Engineered transposases and methods of use thereof

Abstract

The present invention provides engineered transposase polypeptides useful in preparation of next generation sequencing libraries, as well as compositions, methods of utilizing these engineered polypeptides, and polynucleotides encoding the engineered transposases. The present application further relates to methods for using engineered transposases, e.g., in the preparation of nucleic acid libraries for nucleic acid sequencing. More specifically, the invention pertains to methods that use engineered transposases having improved performance to improve the transposition reactions during the preparation of nucleic acid libraries.

Description

ENGINEERED TRANSPOSASES AND METHODS OF USE THEREOFCROSS-REFERENCE TO RELATED APPLICATIONSThis application claims the benefit of U.S. Provisional Application No. 63 / 622,732, filed January 19, 2024, and U.S. Provisional Application No. 63 / 623,064, filed January 19, 2024, both of which are herein incorporated by reference, in their entireties.REFERENCE TO SEQUENCE LISTING, TABLE OR COMPUTER PROGRAMThe instant application contains a Sequence Listing which has been submitted electronically in XML format and is hereby incorporated by reference in its entirety. Said XML copy, created on January 17, 2025, is named “51178-017WO2_Sequence_Listing_1_17_25”, and is 12,833,483 bytes in size.TECHNICAL FIELDThe present invention provides engineered transposase polypeptides useful in the preparation of next generation sequencing (NGS) libraries, as well as compositions and methods of utilizing these engineered polypeptides.BACKGROUNDTransposases are enzymes known to facilitate the transposition of fragments of DNA (transposons) from a donor strand to a target strand at sites flanked by short inverted-repeat sequences or end sequences (ESs). The retroviral integrase superfamily of proteins includes a number of transposases, including Tn5, an IS4 transposase. The transposition mechanism is initiated by the binding of two transposases to donor DNA ESs to form the dimeric synaptic complex or transposome, which catalyzes cleavage of donor DNA. The transposome then transfers the transposon to the target strand (Gradman, R.J., & Reznikoff, W.S. (2008). Tn 5 Synaptic Complex Formation : Role of Transposase Residue W 450.). Two divalent metal ions (Mg2+or Mn2+) bind to the active site and are critical for catalysis (Lovell, S., Goryshin, L, Reznikoff, W. et al. Two-metal active site binding of a Tn 5 transposase synaptic complex. Nat. Struct. Mol. Biol. 9:278-281 (2002).)In nature, transposases have limited activity. However, several mutations in Tn5 are known to enhance transposase activity, including L372P, E54K, E110K, E345K, and P242A / G (Reznikoff, W.S. (2003), Tn5 as a model for understanding DNA transposition. Molecular Microbiology, 47: 1199-1206.) Additionally, a 19 bp ES known as mosaic end (MEs) is associated with hyperactivity in Tn5 transposases.Next generation sequencing (NGS) has facilitated sequencing of large DNA libraries using limited samples. However, sample preparation is time-consuming and resource-intensive, necessitating development of more efficient protocols. Tn5 transposase has been used to reduce costs and improve efficiency in preparing samples for large-scale sequencing projects (Hennig BP, Veiten L, Racks I, Tu CS, Thoms M, Rybin V, Besir H, Remans K, Steinmetz LM. Large-Scale Low-Cost NGS Library PreparationUsing a Robust Tn5 Purification and Tagmentation Protocol. G3 (Bethesda). 2018 Jan 4;8(1):79-89.) In one example, a hyperactive Tn5 with L372P and E54K mutations was used with MEs in a tagmentation protocol to simultaneously fragment sample DNA and bind sequencing adapters for NGS library preparation (Picelli S, Bjorklund AK, Reinius B, Sagasser S, Winberg G, Sandberg R. Tn5 transposase and tagmentation procedures for massively scaled sequencing projects. Genome Res. 2014 Dec;24(12):2033-40.)Improvements to these methods have been reported (Hennig et. al)., and various commercial sample preparation methods now incorporate Tn5 transposase tagmentation. In one instance, a Tn5 transposase is used to create normalized libraries suitable for NGS without the need for independent adjustment or parallelized normalization of input samples or resulting libraries (PCT / US2016 / 013753).However, existing transposase NGS prep methods are limited primarily by their high cost. A transposase with increased DNA fragmentation or cleavage and increased adapter loading or ligation would expand the utility of existing transposase NGS library preparation methods. Transposases with other improved properties, including increased thermostability, increased activity at elevated temperatures, increased soluble expression, decreased inhibition, increased binding to a target polynucleotide or adapter polynucleotide, and / or decreased sequence insertion bias as compared to another wild-type or engineered transposase, are also highly desirable.SUMMARYThe present invention provides engineered transposase polypeptides useful in preparation of next generation sequencing libraries, as well as compositions and methods of utilizing these engineered polypeptides. The transposases of the present invention are derived from a wild-type (WT) polypeptide described by UniProt accession number A0A3L8PTF2 a predicted IS4 family transposase from Parashewanella curva or from a modified hyperactive Tn5 transposase that contains E54K and L372P amino acid substitutions and is described by NCBI Protein accession number HQ908071 .1 . A C- terminal 6-histidine tagged version of the transposases (SEQ ID NO: 2 or SEQ ID NO: 5704, respectively) recognize the 19 bp Tn5 mosaic end (ME) transposon donor recognition sequence. Beneficial properties of these engineered transposases include increased binding to adapters or donor DNA to form a transposome, increased DNA cleavage, increased DNA fragmentation, increased activity at elevated temperatures, increased soluble expression, increased ligation of the adapters or donor DNA to the target DNA, decreased inhibition and / or decreased sequence insertion bias, as compared to a wild-type transposase or a reference transposase.In some embodiments, the present invention provides an engineered transposase polypeptide comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to a reference sequence of SEQ ID NOs: 2, 4, 156, 302, 338, 606, 972, 1082, 1218, 1580, 1598, 1600, 1604, 1720, 2368, 2426, 2540, 3402, 3662, 3788, 4150, 4278, 4642, 4984, 5196, 5286, 5412, 5422, and / or 5704, optionally comprising at least one substitution or one substitution set at one or more positions, wherein the positions are numbered with reference to SEQ ID NOs: 2, 4, 156, 302, 338, 606, 972, 1082, 1218, 1580, 1598, 1600, 1604, 1720, 2368, 2426, 2540, 3402, 3662, 3788, 4150, 4278, 4642, 4984, 5196, 5286, 5412, 5422, and / or 5704, and wherein the engineeredtransposase polypeptide has increased polynucleotide fragmentation or cleavage activity, increased thermostability, increased activity at elevated temperatures, increased soluble expression, decreased inhibition, increased adapter loading or adapter ligation (i.e. tagging), increased binding to a target polynucleotide, and / or decreased sequence insertion bias as compared to another wild-type transposase or engineered transposase. These engineered transposase polypeptides with one or more amino acid substitutions or substitution sets are described, below, in the detailed description of the invention.In some embodiments, the present invention provides engineered transposases comprising a polypeptide sequence having at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to the reference sequence of SEQ ID NOs: 2, 4, 156, 302, 338, 606, 972, 1082, 1218, 1580, 1598, 1600, 1604, 1720, 2368, 2426, 2540, 3402, 3662, 3788, 4150, 4278, 4642, 4984, 5196, 5286, 5412, 5422, and / or 5704, or a functional fragment thereof, wherein the engineered transposase comprises at least one substitution or substitution set in its polypeptide sequence, and wherein the amino acid positions of the polypeptide sequence are numbered with reference to SEQ ID NOs: 2, 4, 156, 302, 338, 606, 972, 1082, 1218, 1580, 1598, 1600, 1604, 1720, 2368, 2426, 2540, 3402, 3662, 3788, 4150, 4278, 4642, 4984, 5196, 5286, 5412, 5422, and / or 5704.In some additional embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 10, 13, 24, 35, 39, 44, 46, 51 , 54, 55, 62 / 361 , 72, 75, 82, 89, 92, 107, 128, 137, 164, 167, 177 / 359, 191 , 192, 193, 194, 201 , 217, 281 , 289, 291 , 291 / 340, 305, 309, 310, 317, 331 , 337, 340, 346, 352, 353, 357, 358, 359, 362, 371 , 396, 414 / 435, 415, 428, 437, 443, 447, and 448, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 2.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 4, 5, 6, 8, 10, 10 / 317, 10 / 317 / 358, 10 / 317 / 359, 10 / 346, 10 / 346 / 359, 10 / 358, 10 / 359, 12, 12 / 373, 13, 15, 16, 18, 19, 26, 30, 30 / 154, 31 , 34, 35, 37, 38, 68, 70, 73, 74, 93, 106, 118, 126, 134, 135, 143, 146, 154 / 453, 163, 182, 207 / 286, 208, 226, 237, 246, 247, 259, 262, 263, 264, 280, 282, 286, 287, 288, 289, 291 , 292, 293, 298, 305, 309, 310, 313, 317, 317 / 346, 317 / 359, 317 / 445, 320, 346, 351 , 355, 356, 358, 359, 360, 368, 369, 410, 411 , 422, 426, 427, 428, 446, 448, 453, and 454, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 4.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 18 / 73 / 74 / 259, 18 / 73 / 286 / 289 / 411 , 18 / 74 / 118 / 259 / 286 / 289 / 411 , 18 / 74 / 259, 18 / 74 / 259 / 286 / 289, 18 / 74 / 259 / 286 / 411 , 18 / 118 / 259, 18 / 259 / 286 / 289 / 411 , 18 / 259 / 289 / 411 , 18 / 289 / 411 , 73 / 74 / 259 / 286 / 289, 73 / 74 / 259 / 289, 73 / 259 / 286, 74 / 259, 74 / 259 / 289, 74 / 286 / 289 / 411 , 259, 259 / 286 / 289, and 289, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 156.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 16 / 19 / 355 / 427, 16 / 37 / 146 / 286 / 287 / 289 / 310 / 313 / 355, 16 / 146 / 226 / 287 / 289 / 355, 19 / 37 / 146 / 286 / 287 / 310 / 355, 19 / 135 / 286 / 287 / 289 / 355, 19 / 182 / 355, 37 / 70 / 226 / 310 / 313 / 355,37 / 74 / 310 / 355 / 427, 37 / 135 / 146 / 182 / 226 / 286 / 287 / 355 / 368, 37 / 146 / 226 / 289 / 427, 37 / 146 / 287 / 289 / 427, 38, 41 , 42, 43, 44, 45, 47, 48, 50, 53, 54, 55, 70 / 74 / 226 / 286 / 287 / 289 / 368 / 427, 70 / 74 / 355 / 368, 70 / 135 / 146 / 150 / 182 / 286 / 289 / 313 / 427, 74 / 310 / 313 / 355, 74 / 310 / 355, 74 / 355 / 427, 101 , 103, 117, 135 / 226 / 286 / 289 / 355 / 368, 135 / 355, 146, 146 / 150 / 287 / 289 / 310 / 355, 146 / 182 / 355, 146 / 226 / 286 / 355, 146 / 286 / 287 / 289 / 355 / 427, 146 / 289, 146 / 427, 150 / 226 / 310 / 313, 182 / 226 / 355, 182 / 355, 244, 247, 249, 251 , 287 / 355 / 427, 300, 317 / 411 , 355, 393, 397, 398, 399, 401 , 405, 407, 408, 409, 411 , 413, 426, 429, 431 , 439, 444, 446, 450, 453, 454, 457, 459, 460, 464, 466, 467, 469, 470, 471 , 472, 474, 475, 476, and 478, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 302.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 16, 16 / 313, 16 / 409 / 411 / 427, 37 / 150 / 408, 37 / 313 / 317 / 408 / 411 , 37 / 317 / 405 / 408, 37 / 317 / 405 / 426, 37 / 405 / 409 / 427, 150, 150 / 313 / 405 / 411 , 150 / 405 / 406, 150 / 408, 150 / 408 / 411 , 226 / 405, 317 / 460, and 408 / 426, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 338.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 12, 13, 18, 19, 22, 26, 31 / 262, 34, 60, 79, 80, 81 , 82, 92, 106, 107, 113, 115, 132, 134, 136,140, 143, 152, 160, 163, 167, 168, 175, 179, 183, 195, 199, 201 , 202, 215, 216, 218, 220, 223, 226, 233,235, 236, 237, 243, 255, 259, 261 , 262, 266, 268, 270, 273, 282, 290, 292, 293, 309, 319, 320, 326, 343,346, 365, 370, 374, 380, 382, 386, 389, 390, 408, 412, 414, 415, 424, and 442, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 606.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 2, 12, 13, 15, 16, 18, 19, 22, 26, 30, 31 / 262, 34, 51 , 60, 69, 79, 82, 84, 87, 88, 106, 107, 113, 114, 115, 128, 132, 134, 135, 136, 140, 143, 146, 150, 152, 160, 163, 167, 168, 175, 179, 183, 195,198, 199, 201 , 215, 217, 218, 219, 220, 223, 224, 226, 227, 231 , 233, 235, 236, 243, 254, 255, 259, 261 ,266, 267, 268, 270, 272, 273, 275, 277, 279, 282, 287, 289, 290, 291 , 292, 293, 304, 307, 309, 319, 320,343, 344, 346, 355, 365, 370, 373, 380, 382, 386, 389, 390, 408, 412, 414, 415, 418, 423, 424, and 468, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 606.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 44 / 45 / 47 / 408 / 453, 44 / 45 / 47 / 453, 44 / 45 / 47 / 453 / 454, 44 / 45 / 103 / 453 / 457, 44 / 45 / 453 / 454 / 457, 44 / 47 / 103 / 393 / 408 / 453 / 454, 44 / 47 / 103 / 453 / 454 / 457, 44 / 47 / 393 / 408 / 453 / 457, 44 / 47 / 393 / 453 / 457, 44 / 103 / 408 / 453 / 454, 45 / 47 / 103 / 393 / 453 / 454 / 457, 45 / 47 / 393 / 453 / 454, 45 / 47 / 408 / 423 / 453 / 457, 45 / 47 / 453, 45 / 103 / 393 / 453 / 454, 45 / 453 / 454, 47 / 103 / 408 / 453 / 454, 47 / 393 / 408 / 453 / 454 / 457, 47 / 393 / 453 / 454, 47 / 408 / 453 / 457, 53 / 54 / 244 / 450, 103 / 393 / 408 / 453 / 457, 244 / 450, 408 / 453 / 454, 446 / 450 / 464, and 453 / 457, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 606.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 38 / 45 / 53, 38 / 45 / 53 / 54 / 476, 38 / 45 / 53 / 244, 38 / 45 / 53 / 470 / 472, 38 / 45 / 54 / 244, 38 / 45 / 54 / 470 / 474 / 478, 38 / 45 / 472 / 476 / 478, 38 / 45 / 476, 38 / 53 / 54, 38 / 53 / 54 / 244, 38 / 53 / 54 / 244 / 472 / 478, 38 / 53 / 472, 38 / 54 / 244 / 470 / 474 / 476 / 478, 38 / 244 / 470 / 472 / 474 / 478, 38 / 478, 45 / 53 / 54 / 244 / 470 / 474 / 476 / 478, 45 / 53 / 54 / 470 / 474 / 476, 45 / 53 / 54 / 472 / 478, 45 / 54 / 244 / 474 / 478, 53, 53 / 54 / 470 / 476 / 478, 53 / 244 / 474 / 478, 54, 54 / 244, 54 / 244 / 472 / 476, 54 / 472, 244, and 474 / 478, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 972.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 22 / 38 / 45 / 107 / 259 / 317 / 412, 22 / 38 / 45 / 259 / 317 / 412, 22 / 107, 22 / 107 / 317 / 412, 22 / 134, 22 / 134 / 220, 22 / 134 / 220 / 317, 22 / 134 / 317 / 412, 22 / 317, 38, 38 / 45 / 81 / 143 / 244, 38 / 53 / 243, 38 / 81 , 38 / 81 / 244, 38 / 107 / 134 / 220, 38 / 132, 38 / 220 / 317, 45 / 81 , 45 / 81 / 243, 45 / 81 / 243 / 343, 45 / 143 / 243 / 270 / 343, 45 / 244 / 270, 53 / 115 / 163 / 243 / 270 / 343, 53 / 143 / 243 / 270 / 343, 53 / 270, 64 / 134 / 220 / 317 / 412, 81 / 132 / 243, 107, 107 / 134 / 317, 115 / 244 / 270, 134 / 259, 143 / 243 / 270, 243, 243 / 270, 244, 244 / 270, 259, 270, and 317, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 972.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 22 / 38 / 45 / 107 / 259 / 317 / 412, 22 / 38 / 45 / 259 / 317 / 412, 22 / 38 / 64 / 134 / 220, 22 / 45 / 220 / 259, 22 / 64 / 107 / 259 / 317 / 412, 22 / 64 / 134 / 220, 22 / 107, 22 / 134, 22 / 134 / 317, 22 / 134 / 317 / 412, 22 / 220, 22 / 220 / 412 / 415, 22 / 259 / 317, 22 / 259 / 317 / 412, 22 / 317, 38, 38 / 45 / 81 / 143 / 244, 38 / 45 / 143 / 270, 38 / 53 / 243, 38 / 81 , 38 / 81 / 143 / 243 / 270, 38 / 81 / 183 / 243 / 270, 38 / 81 / 243, 38 / 81 / 244, 38 / 107 / 134 / 220, 38 / 132, 38 / 220 / 317, 38 / 270 / 343, 45 / 81 , 45 / 81 / 132 / 243 / 270, 45 / 81 / 243, 45 / 143 / 243 / 270 / 343, 45 / 244 / 270, 53 / 143 / 243 / 270 / 343, 53 / 244 / 270, 53 / 270, 64, 64 / 107 / 134, 64 / 107 / 317, 64 / 134, 64 / 134 / 220 / 317 / 412, 81 / 132, 81 / 132 / 243, 107, 107 / 134 / 317, 107 / 412, 115 / 244 / 270, 143 / 243 / 270, 220 / 259, 243, 243 / 270, 244 / 270, 255, 259, and 270, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 972.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 143, 143 / 175 / 272 / 273 / 415, 152 / 168 / 216 / 237, 152 / 168 / 423, 160, 160 / 201 / 272, 160 / 201 / 273 / 415, 160 / 226, 160 / 244 / 272 / 273 / 415, 160 / 415, 168, 168 / 187 / 216 / 255 / 423, 168 / 216 / 237, 168 / 255 / 423, 168 / 290 / 423, 187, 187 / 423, 199 / 201 / 226, 199 / 201 / 415, 201 , 216, 216 / 423, 226 / 244 / 415, 226 / 415, 237, 237 / 290, 244, 255, 272, 290, 290 / 423, 415, and 423, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 1082.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 143, 143 / 160 / 226 / 415, 143 / 175 / 272 / 273 / 415, 152 / 168 / 216 / 237, 152 / 168 / 423, 160, 160 / 201 / 272, 160 / 201 / 273 / 415, 160 / 226, 160 / 244 / 272 / 273 / 415, 160 / 415, 168, 168 / 187 / 216 / 255 / 423, 168 / 216 / 237, 168 / 216 / 237 / 255 / 343, 168 / 255 / 423, 168 / 290 / 423, 187, 199 / 201 / 226, 199 / 201 / 415, 201 ,216, 216 / 423, 226 / 244 / 415, 226 / 415, 237, 237 / 290, 244, 255, 255 / 343, 272, 290, 290 / 423, 415, and 423, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 1082.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 72, 90, 91 , 92, 93, 95, 122, 129, 141 , 142, 143, 144, 145, 150, 151 , 151 / 155, 152, 154, 155, 157, 160, 161 , 161 / 313, 164, 165, 168, 180, 181 , 184, 193, 197, 204, 205, 205 / 472, 206, 299, and 320, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 1082.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 90, 92, 122, 129, 141 , 143, 144, 145, 150, 151 , 151 / 155, 152, 154, 155, 155 / 265, 157, 160, 161 , 164, 165, 168, 180, 181 , 184, 193, 197, 204, 205, 205 / 472, 206, 299, and 320, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 1082.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 129, 129 / 160, 129 / 160 / 164, 129 / 160 / 184, 129 / 164, 129 / 164 / 184, 129 / 205, 146 / 152 / 244, 150 / 155 / 226, 150 / 155 / 244, 152, 154, 154 / 160 / 184, 154 / 164, 154 / 205, 155, 160, 160 / 164, 160 / 164 / 184, 160 / 184, 164, 164 / 204, 164 / 205, 204, 205, 226, 226 / 244, and 244, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 1218.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 129, 129 / 143, 129 / 154 / 164, 129 / 154 / 164 / 205, 129 / 154 / 205, 129 / 160, 129 / 160 / 164, 129 / 160 / 205, 129 / 164 / 205, 143, 143 / 150 / 155 / 226 / 244, 143 / 150 / 155 / 244, 143 / 150 / 226, 143 / 150 / 226 / 244 / 343, 143 / 150 / 226 / 343, 143 / 150 / 244, 143 / 150 / 343, 143 / 152, 143 / 152 / 226, 143 / 152 / 226 / 244, 143 / 152 / 226 / 343, 143 / 152 / 343, 143 / 155, 143 / 155 / 226 / 244, 143 / 155 / 226 / 244 / 343, 143 / 155 / 343, 143 / 226, 143 / 226 / 244 / 343, 143 / 226 / 343, 143 / 244, 143 / 244 / 343, 143 / 343, 146 / 152 / 244, 150 / 155 / 226, 150 / 155 / 244, 150 / 155 / 244 / 343, 150 / 226 / 244 / 343, 152, 152 / 174 / 244 / 343, 152 / 226, 152 / 226 / 343, 152 / 343, 154, 154 / 160 / 184, 154 / 164, 154 / 164 / 204, 154 / 164 / 204 / 205, 154 / 205, 155, 160, 160 / 164, 160 / 164 / 184, 164, 205, 226, 226 / 244, and 244, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 1218.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 98 / 129 / 157 / 168, 129 / 157 / 168, 151 , 151 / 152 / 255, 151 / 152 / 273, 151 / 255, 151 / 343, 157, 157 / 164, 157 / 164 / 168, 157 / 168, 164, 164 / 255, 168, 183, 255, 255 / 343, and 343, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 1580.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 129, 129 / 157, 129 / 157 / 168, 129 / 164, 129 / 164 / 168 / 183, 129 / 164 / 183, 143, 143 / 151 ,143 / 151 / 152, 143 / 151 / 164, 143 / 164, 143 / 164 / 255, 143 / 164 / 343, 143 / 255, 143 / 343, 151 , 151 / 152, 151 / 152 / 164 / 343, 151 / 152 / 343, 151 / 255, 151 / 255 / 343, 151 / 343, 152, 152 / 255, 157, 157 / 164 / 168, 157 / 168, 164, 164 / 168, 164 / 255, 164 / 447, 168, 183, 255, 255 / 343, 298 / 343, and 343, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 1580.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 69, 69 / 164, 72, 73, 91 , 92 / 164, 95, 96, 97, 98, 110, 112, 113, 121 , 121 / 164, 122, 122 / 164, 124, 126, 127, 139, 140, 141 , 144, 144 / 164, 145, 146, 147, 148, 150, 150 / 164, 151 , 151 / 164, 153, 153 / 164, 155, 156, 157, 159, 160, 161 , 163, 164, 164 / 172, 164 / 173, 164 / 182, 164 / 188, 164 / 191 , 164 / 192, 164 / 195, 164 / 197, 164 / 199, 164 / 202, 164 / 205 / 249, 164 / 209, 165, 166, 167, 169, 170, 171 , 172, 173, 177, 184, 187, 188, 190, 191 , 192, 193, 194, 195, 197, 197 / 266, 198, 199, 200, 202, 203, 204, 205, 208, 209, 210, 321 , 324, 331 , and 349, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 1580.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 69, 69 / 164, 72, 73, 91 , 92 / 164, 95, 97, 98, 121 / 164, 126, 127, 139, 140, 142, 144, 144 / 164, 145, 146, 146 / 164, 148, 150, 150 / 164, 151 , 153, 153 / 164, 155, 156, 157, 160, 163, 164 / 173, 164 / 188, 164 / 191 , 164 / 192, 164 / 195, 164 / 199, 164 / 205 / 249, 164 / 209, 165, 166, 167, 169, 171 , 172, 173, 177, 182, 187, 188, 190, 191 , 192, 194, 195, 197 / 266, 198, 199, 200, 202, 203, 204, 205, 208, 209, 210, 324, 331 , and 349, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 1580.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 73, 91 , 93, 95, 97, 98, 110, 112, 122, 122 / 164, 124, 127, 139, 144, 145, 146, 146 / 164, 147, 148, 150, 150 / 164, 151 , 151 / 164, 153, 153 / 164, 155, 156, 157, 159, 160, 161 , 162, 163, 164, 164 / 182, 164 / 188, 164 / 191 , 164 / 192, 164 / 195, 164 / 199, 164 / 205 / 249, 164 / 209, 165, 166, 167, 169, 171 , 172, 177, 184, 187, 188, 190, 191 , 192, 193, 195, 197 / 266, 198, 199, 200, 204, 205, 206, 207, 208, 209, 210, and 324, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 1580.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 69, 73, 91 , 127, 140, 144, 145, 146, 146 / 164, 150, 150 / 164, 151 , 153, 155, 156, 157, 160, 163, 164, 164 / 172, 164 / 188, 164 / 191 , 164 / 192, 164 / 195, 164 / 199, 164 / 209, 165, 167, 169, 171 , 172, 182, 183, 188, 191 , 192, 193, 195, 196, 198, 199, 200, 204, 205, 208, 210, and 324, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 1580.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 38 / 143 / 168 / 220 / 290 / 317 / 423, 38 / 152 / 154 / 164 / 168 / 220 / 290 / 317 / 423, 38 / 152 / 168 / 220 / 290 / 317 / 343 / 423, 38 / 152 / 168 / 220 / 290 / 317 / 423, 38 / 154 / 164 / 168 / 220 / 290 / 317 / 423,38 / 168 / 220 / 290 / 317 / 423, and 38 / 220 / 317, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 972.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 73 / 143 / 169 / 191 / 193, 73 / 156, 73 / 156 / 192 / 435, 73 / 156 / 210, 73 / 163, 73 / 163 / 210, 73 / 192 / 193, 127, 127 / 160 / 172 / 198, 127 / 172, 127 / 198, 146, 146 / 151 / 188, 146 / 324, 151 / 171 / 324, 151 / 324, 156 / 169, 156 / 192, 157 / 171 , 163, 163 / 169, 163 / 169 / 191 / 192 / 193, 165 / 171 / 324, 165 / 188 / 324, 165 / 324, 169, 171 / 188 / 324, 171 / 324, 172, 188, 192, 192 / 193, 198, 210, and 324 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 2368.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 73, 73 / 163 / 191 / 192 / 193, 127 / 160 / 172, 127 / 160 / 172 / 198, 144, 160, 160 / 172, 163 / 191 / 192, and 192 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 2368. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 2396. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 2396, wherein the C-terminal histidine tag is replaced or removed. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 2396, wherein the C-terminal peptide spacer GGSG (SEQ ID NO: 5777) or a portion thereof is replaced or removed.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 8, 13, 17, 18, 20 / 146 / 157 / 167, 26, 34 / 38, 38 / 39 / 45, 67, 70, 73, 73 / 150 / 155 / 169, 73 / 150 / 171 / 192, 73 / 155, 73 / 155 / 169 / 199, 73 / 155 / 171 , 73 / 192, 73 / 199, 78, 82, 84, 86, 86 / 384, 89, 90, 91 , 95, 102, 110, 111 , 113, 114, 115, 122 / 127, 126 / 127, 127 / 131 / 165, 127 / 133, 143, 143 / 146, 143 / 146 / 153, 143 / 146 / 157, 143 / 146 / 167, 143 / 146 / 167 / 188, 143 / 146 / 188, 143 / 153 / 167 / 188 / 246, 143 / 156 / 157 / 167 / 195, 143 / 156 / 188, 143 / 188, 143 / 188 / 195, 143 / 195, 144, 145, 145 / 151 , 146, 146 / 152, 146 / 153 / 157, 146 / 153 / 157 / 167, 146 / 153 / 167, 146 / 157, 146 / 157 / 167, 146 / 167, 150, 150 / 155 / 169, 150 / 155 / 169 / 192, 150 / 155 / 192, 150 / 169, 150 / 192, 150 / 199, 151 / 152 / 154, 152 / 154, 152 / 154 / 155 / 160, 153 / 157 / 167, 155 / 171 , 155 / 192, 156 / 270, 160 / 371 , 167 / 188, 168 / 172, 169, 184, 190, 192 / 198, 198, 206, 208, 209, 210, 214 / 220, 215 / 220, 216 / 220, 217 / 220, 219 / 220, 220, 220 / 222, 220 / 225, 220 / 226, 231 , 235, 242, 257, 259, 261 , 262, 267, 268, 269, 270, 272, 274, 275, 276, 277, 278, 282, 283 / 290, 284 / 290, 287 / 290, 288 / 290, 289 / 290, 290, 290 / 291 , 290 / 292, 290 / 293, 290 / 294, 304, 305, 305 / 373, 306, 307, 309, 317, 329, 333, 334, 339, 341 , 344, 345, 346, 352, 353, 355, 358, 366, 368, 370, 372, 373, 375, 376, 378, 379, 380, 381 , 382, 383, 384, 385, 386 / 393, 387 / 393, 389 / 393, 390 / 393, 391 / 393, 392 / 393, 393 / 394, 414, 415, 420 / 423, 422 / 423, 423, 423 / 425, 432, 440, 441 , 443, 469, and 472 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 2426.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 8, 18, 25, 29, 30, 67, 73, 73 / 150 / 155 / 169, 73 / 150 / 155 / 171 , 73 / 150 / 171 / 192, 73 / 171 , 73 / 192, 73 / 199, 74, 82, 86, 86 / 384, 113, 115, 118, 127 / 128, 143, 150, 150 / 155 / 169, 150 / 155 / 169 / 192,150 / 155 / 171 , 150 / 155 / 171 / 199, 150 / 155 / 192, 150 / 155 / 199, 150 / 192, 150 / 199, 155 / 192, 168 / 171 / 172, 195 / 198, 197 / 198, 198 / 201 , 198 / 203, 198 / 205, 235, 252, 256, 257, 262, 263, 267 / 343, 268, 269, 270, 272, 273, 276, 277, 288 / 290, 289 / 290, 290, 290 / 291 , 290 / 292, 290 / 294, 298, 304, 305, 307, 333, 346, 355, 366, 370, 386 / 393, 389 / 393, 393 / 395, 440, and 472 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 2426.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 66, 67, 70, 101 , 102, 103, 104, 106, 107, 108, 113, 114, 115, 117, 118, 143, 152, 154, 235, 236 / 251 , 237, 238, 242, 243, 246, 247, 249, 250, 251 , 256 / 352, 257, 259, 285, 286, 287, 288, 289, 290, 292, 293, 294, 295, 296, 322, 323, 326, 329, 330, 333, 339, 340, 343, 344, 345, and 352. and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 2540.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 18, 18 / 113 / 270, 18 / 113 / 270 / 329 / 370, 18 / 113 / 270 / 370 / 384, 18 / 113 / 270 / 370 / 412, 18 / 113 / 329 / 384, 18 / 259 / 370 / 384, 18 / 270, 18 / 270 / 370, 18 / 270 / 370 / 384, 18 / 370, 18 / 370 / 384, 73, 73 / 115, 73 / 115 / 184 / 333, 73 / 115 / 184 / 341 , 73 / 115 / 256, 73 / 184 / 256, 73 / 184 / 273 / 333, 73 / 184 / 333, 73 / 256, 73 / 256 / 375, 73 / 273, 73 / 273 / 333 / 375, 73 / 273 / 375, 73 / 333, 73 / 333 / 375, 73 / 375, 73 / 379, 77 / 233 / 238 / 376, 77 / 233 / 334 / 376, 77 / 262 / 376, 77 / 276, 77 / 334 / 376, 82, 82 / 127, 82 / 127 / 144 / 235 / 290, 82 / 127 / 144 / 266, 82 / 127 / 144 / 266 / 290, 82 / 127 / 144 / 378, 82 / 127 / 235, 82 / 127 / 290, 82 / 144 / 266 / 290 / 378, 82 / 144 / 266 / 378, 82 / 235 / 266, 82 / 235 / 266 / 378 / 383, 82 / 235 / 290, 82 / 235 / 378, 82 / 266, 82 / 266 / 378, 82 / 378, 113 / 270 / 370 / 384, 115 / 256 / 333 / 375, 115 / 256 / 375, 115 / 273 / 375, 115 / 375, 127 / 144 / 290, 127 / 149 / 150, 127 / 266 / 290 / 378, 127 / 378 / 383, 144 / 235 / 290, 144 / 266 / 290, 233 / 334, 233 / 376, 235, 235 / 290, 256 / 333, 262, 266 / 378, 270 / 370, 273, 276 / 352, 290, 290 / 378, 333, 358, 370, 375, and 378 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 2540.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 18, 18 / 113, 18 / 113 / 252, 18 / 113 / 252 / 336 / 384, 18 / 113 / 252 / 370, 18 / 113 / 270, 18 / 113 / 270 / 370 / 384, 18 / 259 / 370 / 384, 18 / 270, 18 / 270 / 370, 18 / 270 / 370 / 384, 18 / 370, 18 / 370 / 384, 73, 73 / 115 / 184 / 341 , 73 / 115 / 341 , 73 / 131 / 256 / 273 / 333 / 341 / 375, 73 / 184 / 256, 73 / 184 / 256 / 341 , 73 / 184 / 256 / 375, 73 / 184 / 273 / 333, 73 / 184 / 333, 73 / 184 / 341 / 375, 73 / 256, 73 / 256 / 273, 73 / 256 / 273 / 375, 73 / 256 / 333 / 375, 73 / 256 / 375, 73 / 273, 73 / 273 / 333 / 341 , 73 / 273 / 333 / 375, 73 / 273 / 375, 73 / 333, 73 / 333 / 341 / 375, 73 / 333 / 375, 73 / 341 , 73 / 375, 73 / 379, 77, 77 / 118, 77 / 118 / 143 / 262 / 276 / 334 / 376, 77 / 118 / 143 / 334 / 352 / 376, 77 / 118 / 143 / 376, 77 / 118 / 233, 77 / 118 / 233 / 352 / 376, 77 / 118 / 262 / 334 / 376, 77 / 118 / 334, 77 / 143 / 262 / 276, 77 / 143 / 262 / 334, 77 / 143 / 376, 77 / 233 / 238 / 376, 77 / 233 / 262, 77 / 233 / 276 / 376, 77 / 233 / 334 / 376, 77 / 262 / 376, 77 / 276, 77 / 334 / 376, 77 / 352, 82, 82 / 127, 82 / 127 / 144 / 235 / 290, 82 / 127 / 144 / 266, 82 / 127 / 144 / 266 / 290, 82 / 127 / 144 / 378, 82 / 127 / 235, 82 / 127 / 235 / 266 / 358, 82 / 127 / 290, 82 / 127 / 290 / 358, 82 / 127 / 290 / 378, 82 / 144 / 266 / 290 / 378, 82 / 144 / 266 / 378, 82 / 150 / 235 / 266, 82 / 235 / 266, 82 / 235 / 266 / 290, 82 / 235 / 266 / 378 / 383, 82 / 235 / 290, 82 / 235 / 378, 82 / 266, 82 / 266 / 378, 82 / 290, 82 / 378, 113, 113 / 252, 113 / 252 / 270 / 370, 113 / 270 / 370 / 384, 113 / 336, 118 / 233 / 276 / 334, 118 / 334, 127 / 235 / 266, 127 / 266 / 290 / 378, 127 / 378 / 383, 143,143 / 276, 143 / 276 / 334, 144 / 235 / 290, 144 / 266 / 290, 144 / 290 / 358, 233 / 334, 233 / 376, 235, 235 / 266 / 290 / 358 / 383, 235 / 358, 252 / 329 / 336 / 370, 252 / 384, 256 / 333, 256 / 375, 262, 262 / 276, 262 / 334, 266 / 290, 266 / 378, 270 / 370, 273, 276 / 352, 290, 290 / 378, 333, 334, 334 / 376, 336 / 370, 341 , 358, 370, 375, and 378 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 2540.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 18, 18 / 77 / 118 / 169 / 233 / 238 / 256 / 266 / 270, 18 / 77 / 150 / 256 / 376, 18 / 118, 18 / 118 / 143 / 169 / 238 / 256 / 266 / 333, 18 / 118 / 150 / 169 / 184 / 238 / 256 / 270 / 378, 18 / 118 / 169, 18 / 118 / 233 / 238, 18 / 143, 18 / 143 / 256, 18 / 150 / 169, 18 / 150 / 266, 18 / 169 / 184 / 290 / 333 / 376, 18 / 169 / 233 / 333, 18 / 169 / 392, 18 / 184 / 270, 18 / 233 / 256 / 334, 18 / 256 / 266 / 290 / 378, 77 / 113 / 118 / 256 / 266 / 290, 77 / 118 / 290, 77 / 169 / 270 / 290, 77 / 233, 77 / 233 / 256, 77 / 238, 113 / 118 / 169 / 184 / 256 / 376, 113 / 169, 113 / 233, 118, 118 / 143 / 238, 118 / 233 / 238 / 256 / 378, 118 / 233 / 238 / 270, 143 / 169 / 233 / 270, 143 / 233 / 266 / 290 / 376, 143 / 238 / 270 / 290 / 378, 143 / 256 / 334, 143 / 290, 169, 169 / 256 / 266, 183 / 270 / 333, 233 / 238, 233 / 238 / 376, 233 / 392, 238 / 256, 256 / 290 / 303, 270, 290, 376, and 378 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 3402.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 18, 18 / 77 / 118 / 169 / 266 / 334 / 376, 18 / 77 / 150 / 256 / 376, 18 / 77 / 256 / 290 / 334, 18 / 113 / 238, 18 / 118, 18 / 118 / 143 / 169 / 238 / 256 / 266 / 333, 18 / 118 / 143 / 169 / 376, 18 / 118 / 169, 18 / 118 / 233 / 238, 18 / 118 / 266, 18 / 143, 18 / 143 / 169 / 233 / 238 / 358, 18 / 143 / 233 / 238 / 290 / 376, 18 / 143 / 256, 18 / 143 / 290, 18 / 143 / 333 / 334 / 378, 18 / 150 / 169, 18 / 150 / 266, 18 / 169 / 184 / 290 / 333 / 376, 18 / 169 / 233 / 333, 18 / 169 / 238 / 270 / 334 / 376, 18 / 169 / 392, 18 / 233 / 256 / 334, 18 / 233 / 266 / 290 / 376, 18 / 256 / 266 / 290 / 378, 18 / 266 / 270 / 378, 18 / 378, 77 / 113 / 121 / 143 / 233 / 334, 77 / 118 / 169 / 233 / 266 / 270, 77 / 118 / 238 / 376, 77 / 118 / 290, 77 / 169 / 270 / 290, 77 / 233, 77 / 238, 113 / 238 / 376, 118, 118 / 143 / 238, 118 / 143 / 392, 118 / 169 / 184 / 233 / 266 / 270 / 333, 118 / 233 / 238 / 256 / 378, 118 / 256 / 290 / 333 / 334 / 376 / 378, 118 / 256 / 334 / 376, 118 / 266 / 290, 143 / 169 / 233 / 270, 143 / 233 / 266 / 290 / 376, 143 / 238 / 270 / 290 / 378, 143 / 256 / 334, 143 / 290, 143 / 364, 150 / 233 / 333, 169, 169 / 256 / 266, 183 / 270 / 333, 233 / 238, 233 / 238 / 376, 233 / 392, 238 / 256, 238 / 266 / 270 / 378, 256 / 266 / 334, 266 / 270 / 376, 266 / 333, and 378 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 3402.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 77 / 156, 77 / 156 / 270, 77 / 184 / 244, 77 / 184 / 333, 77 / 244 / 270, 77 / 270, 143, 143 / 169 / 238 / 257 / 344, 143 / 169 / 255, 143 / 169 / 255 / 257, 143 / 169 / 343, 143 / 255, 143 / 255 / 257, 143 / 255 / 344, 143 / 257, 156, 156 / 270, 169 / 238, 169 / 238 / 255 / 257 / 344, 169 / 257 / 344, 169 / 344, 184, 184 / 244 / 324, 238, 238 / 255 / 344, 244 / 270, 257 / 344, and 270 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 3662.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 77, 77 / 156 / 184 / 244 / 324, 77 / 184 / 270, 143, 143 / 238, 143 / 238 / 255, 143 / 238 / 255 / 343,143 / 238 / 255 / 392, 143 / 238 / 344, 143 / 255, 143 / 257 / 392, 143 / 344, 169, 169 / 238 / 255 / 344, 169 / 238 / 344, 169 / 255, 169 / 343, 169 / 392, 238 / 344, 255, 255 / 257, 255 / 257 / 392, 257, and 344 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 3662.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 20, 22, 26, 42, 43, 44, 45, 50, 51 , 52, 53, 54, 55, 60, 62, 64, 65, 95, 97, 99, 103, 104, 111 , 112, 115, 115 / 143 / 169, 115 / 169 / 343, 115 / 343, 116, 116 / 343, 143, 143 / 169, 154 / 439, 169, 169 / 324, 169 / 324 / 343, 169 / 343, 186, 234, 236, 238, 238 / 246, 239, 239 / 244, 241 , 242, 243, 244, 250, 251 , 254, 255, 260, 283, 284, 295, 296, 298 / 326, 319, 322, 323, 324, 324 / 343, 326, 328, 329, 333, 334, 338, 340, 341 , 343, 344, 347, 366, 434, 439, 442, 443, 444, 445, 446, and 447 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 3788.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 26, 26 / 236, 26 / 236 / 260 / 333, 26 / 236 / 295, 26 / 260, 26 / 260 / 295, 26 / 260 / 295 / 333, 26 / 295 / 319, 26 / 333, 64, 64 / 244, 64 / 251 / 260 / 347, 64 / 260 / 295 / 333, 64 / 260 / 347, 103 / 115 / 255, 103 / 115 / 343, 103 / 115 / 343 / 344, 103 / 143 / 255 / 284 / 343 / 344, 103 / 169 / 343 / 344, 103 / 255 / 284, 103 / 343 / 344, 115, 115 / 143 / 255, 115 / 143 / 255 / 343, 115 / 143 / 284, 115 / 143 / 343, 115 / 143 / 344, 115 / 169, 115 / 169 / 343, 115 / 255, 115 / 255 / 344, 115 / 284, 115 / 284 / 343 / 344, 115 / 284 / 344, 115 / 343, 115 / 343 / 344, 115 / 344, 143, 143 / 169 / 255 / 284 / 343 / 344, 143 / 169 / 343 / 344, 143 / 255, 143 / 255 / 284 / 343 / 344, 143 / 255 / 343, 143 / 255 / 343 / 344, 143 / 255 / 344, 143 / 343 / 344, 169, 169 / 255, 169 / 255 / 284, 169 / 255 / 343, 169 / 284, 169 / 343, 169 / 344, 236, 236 / 244 / 333, 236 / 260, 236 / 347, 244, 244 / 251 , 244 / 251 / 319, 244 / 260, 244 / 295, 244 / 347, 251 , 251 / 260 / 295 / 347, 251 / 333, 255, 255 / 284, 255 / 343, 255 / 343 / 344, 255 / 344, 259, 260, 260 / 295, 260 / 333, 284, 284 / 343 / 344, 284 / 344, 295, 295 / 333, 333, 343, 343 / 344, 344, and 347 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 4150.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 115, 115 / 117, 115 / 117 / 118, 115 / 117 / 118 / 152, 115 / 117 / 118 / 169, 115 / 117 / 118 / 246, 115 / 117 / 152, 115 / 117 / 152 / 169, 115 / 117 / 152 / 169 / 246, 115 / 117 / 152 / 246, 115 / 117 / 169, 115 / 117 / 246, 115 / 118, 115 / 118 / 152, 115 / 118 / 152 / 169, 115 / 118 / 152 / 169 / 246, 115 / 118 / 169, 115 / 118 / 246, 115 / 152, 115 / 152 / 169, 115 / 152 / 169 / 246, 115 / 152 / 246, 115 / 152 / 246 / 374, 115 / 169, 115 / 169 / 246, 115 / 246, 117, 117 / 118, 117 / 118 / 152, 117 / 118 / 152 / 169, 117 / 118 / 152 / 246, 117 / 118 / 169, 117 / 118 / 246, 117 / 152, 117 / 152 / 169, 117 / 152 / 246 / 288, 117 / 169, 117 / 246, 118 / 246, 152, 152 / 169, 152 / 169 / 246, 152 / 246, 169, 169 / 246, and 246 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 4278. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 4642. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 4642, wherein the C-terminal histidine tag is replaced or removed. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 4642, wherein the C-terminal peptide spacer GGSG (SEQ ID NO: 5777) or a portion thereof is replaced or removed.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 73, 101 , 114, 115, 118, 143, 145, 150, 151 , 152, 153, 154, 156, 157, 160, 161 , 163, 166, 167, 168, 169, 171 , 172, 190, 191 , 192, 195, 213, 214, 218, 219, 223, 225, 233, 233 / 343, 234, 235, 236, 237, 238, 239, 239 / 250, 243, 244, 249, 251 , 254, 255, 257, 258, 259, 261 , 262, 262 / 283, 277 / 292, 283, 284, 286, 289, 292, 295, and 296 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 4278.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 93, 113, 113 / 114, 113 / 115, 113 / 143, 115 / 118, 115 / 177, 118 / 143 / 163 / 168 / 195 / 255 / 315, 118 / 143 / 255, 118 / 163 / 168 / 214 / 255 / 315, 118 / 168 / 214 / 315, 118 / 255, 124, 126, 129, 134, 143 / 154 / 255, 143 / 156 / 168 / 216 / 255, 143 / 163, 143 / 163 / 255, 143 / 168, 143 / 168 / 169 / 255 / 315, 143 / 255, 148, 156 / 214 / 315, 159, 163, 163 / 168, 163 / 168 / 169, 163 / 168 / 169 / 255 / 315, 163 / 168 / 169 / 315, 163 / 168 / 195 / 214 / 255 / 315, 163 / 214, 163 / 255, 163 / 255 / 315, 168 / 315, 186, 194, 197, 206, 207, 208, 210, 213, 255 / 315, 277, 293, 294, 315, 317, 320, 322, 328, 329, 330, 333, 352, 366, 392, and 445 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 4642. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 4872. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 4872, wherein the C-terminal histidine tag is replaced or removed. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 4872, wherein the C-terminal peptide spacer GGSG (SEQ ID NO: 5777) or a portion thereof is replaced or removed. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 4954. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 4954, wherein the C-terminal histidine tag is replaced or removed. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 4954, wherein the C-terminal peptide spacer GGSG (SEQ ID NO: 5777) or a portion thereof is replaced or removed.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 127, 128, 129, 140, 180, 182, 194, 277, 293, 294, 299, 300, 317, 319, 320, 324, 333, 356, 366, and 392 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 4642.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 113, 117, 118, 151 , 191 , 196, and 205 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 4984.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 114 / 115 / 143, 114 / 115 / 143 / 233, 114 / 115 / 233, 114 / 115 / 343, 114 / 143, 114 / 143 / 151 / 233 / 238 / 261 , 114 / 143 / 213 / 261 / 343, 114 / 143 / 233, 114 / 143 / 233 / 238, 114 / 151 / 261 / 343, 114 / 213 / 238, 114 / 343, 115, 115 / 143, 115 / 143 / 151 / 154 / 213 / 261 , 115 / 143 / 151 / 154 / 233 / 343,115 / 143 / 151 / 233 / 261 , 115 / 143 / 151 / 233 / 343, 115 / 143 / 154 / 213 / 233 / 238 / 261 , 115 / 143 / 213, 115 / 143 / 233, 115 / 143 / 343, 115 / 151 / 154 / 233, 115 / 151 / 261 , 115 / 213 / 233, 115 / 213 / 233 / 238, 115 / 213 / 261 , 115 / 233 / 238, 115 / 238, 115 / 343, 115 / 348, 143, 143 / 151 / 154, 143 / 151 / 154 / 213, 143 / 154, 143 / 213 / 233, 143 / 213 / 233 / 238 / 261 / 343, 143 / 213 / 261 / 343, 143 / 233 / 261 , 143 / 261 , 151 / 154, 151 / 154 / 213 / 343, 151 / 154 / 343, 151 / 213, 151 / 213 / 233, 213 / 233 / 343, and 233 / 238 / 343 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 4984. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 5186. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 5186, wherein the C-terminal histidine tag is replaced or removed. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 5186, wherein the C-terminal peptide spacer GGSG (SEQ ID NO: 5777) or a portion thereof is replaced or removed. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 5354. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 5354, wherein the C-terminal histidine tag is replaced or removed. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 5354, wherein the C-terminal peptide spacer GGSG (SEQ ID NO: 5777) or a portion thereof is replaced or removed.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 117, 117 / 118 / 143 / 168 / 169 / 392, 117 / 118 / 143 / 392, 117 / 118 / 168 / 169, 117 / 118 / 168 / 169 / 392, 117 / 143 / 168 / 169 / 170 / 392, 117 / 143 / 168 / 169 / 392, 117 / 168 / 169, 117 / 168 / 169 / 392, 117 / 168 / 392, 117 / 169, 117 / 392, 118 / 143, 118 / 143 / 168 / 169, 118 / 143 / 169, 118 / 143 / 392, 118 / 168, 118 / 168 / 392, 118 / 169 / 392, 118 / 392, 143 / 168 / 169 / 392, 143 / 169, 143 / 392, 168 / 169, 168 / 169 / 392, 168 / 392, 169 / 392, and 392 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 5196.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 134, 134 / 182 / 194 / 294, 134 / 182 / 277 / 293, 134 / 182 / 293 / 320, 134 / 182 / 294, 134 / 194 / 277, 134 / 194 / 277 / 317, 134 / 194 / 293, 134 / 194 / 293 / 294, 134 / 194 / 293 / 317, 134 / 194 / 294, 134 / 194 / 317, 134 / 194 / 317 / 320, 134 / 317, 182, 182 / 194, 182 / 194 / 293 / 320, 182 / 293 / 294, 194, 194 / 277 / 293 / 317, 194 / 277 / 317 / 320, 194 / 277 / 320, 194 / 293 / 294 / 317, 194 / 293 / 320, 277, 293, 317, 317 / 320, and 320 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 5286.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 12 / 134, 13 / 134, 13 / 134 / 210, 22 / 134 / 152, 22 / 134 / 235 / 344, 64 / 93 / 134 / 235, 64 / 134 / 344 / 347, 77 / 134, 79 / 134 / 210, 84 / 134, 86 / 134, 87 / 134, 89 / 134, 93 / 134 / 152 / 235 / 344, 93 / 134 / 235 / 344, 93 / 134 / 244 / 344, 93 / 134 / 344, 131 , 134 / 152 / 344, 134 / 175, 134 / 210 / 382, 134 / 210 / 402, 134 / 210 / 456, 134 / 210 / 478, 134 / 344, 134 / 377, 134 / 378, 134 / 380, 134 / 381 , 134 / 382, 134 / 384, 134 / 386, 134 / 387, 134 / 388, 134 / 390, 134 / 391 , 134 / 393, 134 / 395, 134 / 398, 134 / 401 , 134 / 402, 134 / 406, 134 / 407, 134 / 421 , 134 / 456, 134 / 457, 134 / 459, 134 / 460, 134 / 464, 134 / 467, 134 / 468, 134 / 470, 134 / 471 , 134 / 473,134 / 473 / 474, 134 / 474, and 134 / 475 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 5412.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 73 / 127 / 134 / 160 / 172 / 198 / 290, 73 / 127 / 134 / 160 / 198 / 333, 73 / 127 / 134 / 160 / 210 / 266, 73 / 127 / 134 / 172 / 192 / 266 / 333, 73 / 127 / 134 / 172 / 198, 73 / 127 / 134 / 210 / 290, 73 / 127 / 134 / 210 / 333, 73 / 127 / 134 / 290 / 333, 73 / 127 / 134 / 333, 73 / 134, 73 / 134 / 160 / 172 / 192 / 290, 73 / 134 / 160 / 172 / 198 / 333, 73 / 134 / 160 / 172 / 290, 73 / 134 / 160 / 266, 73 / 134 / 160 / 333, 73 / 134 / 172, 73 / 134 / 172 / 210 / 333, 73 / 134 / 172 / 266 / 333, 73 / 134 / 198 / 210, 73 / 134 / 198 / 210 / 266, 73 / 134 / 210 / 333, 73 / 134 / 333, 73 / 134 / 366, 127 / 134, 127 / 134 / 160 / 172 / 192 / 333, 127 / 134 / 160 / 172 / 210 / 266 / 290, 127 / 134 / 172 / 198 / 266 / 333, 127 / 134 / 192 / 210, 127 / 134 / 198 / 210 / 290, 127 / 134 / 210 / 266 / 290, 127 / 134 / 210 / 333, 134, 134 / 160 / 172 / 333, 134 / 160 / 198 / 266 / 290, 134 / 172 / 192 / 290 / 333, 134 / 192 / 198, 134 / 198, 134 / 198 / 266 / 333, 134 / 210, 134 / 210 / 290, and 134 / 333 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 5412. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 5500. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 5500, wherein the C-terminal histidine tag is replaced or removed. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 5500, wherein the C-terminal peptide spacer GGSG (SEQ ID NO: 5777) or a portion thereof is replaced or removed. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 5520. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 5520, wherein the C-terminal histidine tag is replaced or removed. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 5520, wherein the C-terminal peptide spacer GGSG (SEQ ID NO: 5777) or a portion thereof is replaced or removed.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from 13, 13 / 178 / 381 , 13 / 178 / 381 / 473, 13 / 178 / 457, 13 / 381 , 13 / 381 / 457, 13 / 402 / 457, 13 / 457, 178, 178 / 381 / 457, 178 / 381 / 457 / 473, 178 / 402 / 457, 178 / 473, 381 , 381 / 393 / 457, 381 / 457 / 473, 381 / 473, and 393 / 457 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 5422.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from the reference sequence is SEQ ID NO: 5704 and the at least one substitution comprises substitutions at amino acid positions selected from 48, 116 ,118, 120, 130, 137, 153, 157, 163, 175, 181 , 187, 192, 213, 246, 263, 273, 296, 318, 334, 341 , 394, 412, 424, 454, and 458 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 5704. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 5738. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 5738, wherein the C-terminal histidine tag is replaced or removed. In some embodiments, the engineeredtransposase comprises the polypeptide sequence of SEQ ID NO: 5738, wherein the C-terminal peptide spacer GGSG (SEQ ID NO: 5777) or a portion thereof is replaced or removed.In some further embodiments, the engineered transposase comprises a polypeptide sequence comprising at least one substitution or substitution set comprising substitutions at amino acid positions selected from38 / 73 / 113 / 115 / 117 / 118 / 127 / 134 / 152 / 154 / 160 / 163 / 164 / 168 / 169 / 172 / 184 / 192 / 194 / 198 / 220 / 243 / 260 / 266 / 27 0 / 295 / 317 / 333 / 392 / 423,38 / 73 / 113 / 115 / 117 / 118 / 127 / 134 / 152 / 154 / 160 / 163 / 164 / 168 / 169 / 172 / 184 / 192 / 198 / 220 / 243 / 260 / 266 / 270 / 29 5 / 317 / 333 / 392 / 423,38 / 73 / 113 / 115 / 117 / 118 / 127 / 134 / 152 / 154 / 160 / 163 / 164 / 168 / 169 / 172 / 184 / 192 / 198 / 220 / 243 / 260 / 266 / 270 / 29 5 / 317 / 333 / 392 / 423 / 457,38 / 73 / 113 / 115 / 117 / 118 / 127 / 152 / 154 / 160 / 163 / 164 / 168 / 169 / 172 / 184 / 192 / 198 / 220 / 243 / 260 / 266 / 270 / 295 / 31 7 / 333 / 392 / 423,38 / 73 / 113 / 115 / 118 / 127 / 152 / 154 / 160 / 162 / 164 / 168 / 172 / 184 / 192 / 198 / 220 / 243 / 260 / 266 / 270 / 295 / 317 / 333 / 42 3,38 / 73 / 113 / 115 / 118 / 127 / 152 / 154 / 160 / 163 / 164 / 168 / 169 / 172 / 184 / 192 / 198 / 220 / 243 / 260 / 266 / 270 / 295 / 317 / 33 3 / 423,38 / 73 / 113 / 115 / 118 / 127 / 152 / 154 / 160 / 164 / 168 / 172 / 184 / 189 / 192 / 198 / 220 / 243 / 260 / 266 / 270 / 295 / 317 / 333 / 42 3,38 / 73 / 113 / 115 / 118 / 127 / 152 / 154 / 160 / 164 / 168 / 172 / 184 / 192 / 198 / 210 / 220 / 243 / 260 / 266 / 270 / 295 / 317 / 333 / 42 3, 38 / 73 / 113 / 115 / 118 / 127 / 152 / 154 / 160 / 164 / 168 / 172 / 184 / 192 / 198 / 220 / 243 / 260 / 266 / 270 / 295 / 317 / 333 / 423, 38 / 73 / 113 / 118 / 127 / 152 / 154 / 160 / 164 / 168 / 172 / 184 / 192 / 198 / 220 / 243 / 260 / 266 / 270 / 295 / 317 / 333 / 423, 38 / 73 / 113 / 118 / 127 / 152 / 154 / 160 / 164 / 168 / 172 / 184 / 192 / 198 / 220 / 243 / 266 / 270 / 317 / 333 / 423, 38 / 73 / 113 / 118 / 127 / 152 / 154 / 160 / 164 / 168 / 172 / 184 / 192 / 198 / 220 / 266 / 270 / 317 / 333 / 423, and 38 / 113 / 127 / 152 / 154 / 160 / 164 / 168 / 172 / 198 / 220 / 290 / 317 / 423, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 972. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 5702. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 5702, wherein the C-terminal histidine tag is replaced or removed. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 5702, wherein the C-terminal peptide spacer GGSG (SEQ ID NO: 5777) or a portion thereof is replaced or removed.In some additional embodiments, the present invention provides engineered transposases capable of binding to adapters or donor DNA to form a transposome, cleaving or fragmenting target DNA, and / or ligating the adapters or donor DNA to the target DNA.In some further embodiments, the present invention provides engineered transposases have at least one improved property, as compared to a wild-type or reference transposase. In some embodiments, the improved property is selected from increased polynucleotide fragmentation or cleavage activity, increased thermostability, increased activity at elevated temperatures, increased solubleexpression, decreased inhibition, increased adapter loading or adapter ligation (i.e. tagging), increased binding to a target polynucleotide, and / or decreased sequence insertion bias.In still some additional embodiments, the present invention provides engineered transposases comprise increased polynucleotide fragmentation or cleavage activity as compared to a wild-type or reference transposase.In yet some additional embodiments, the engineered transposases comprise increased thermal stability as compared to a wild-type or reference transposase. In some further embodiments, the engineered transposase comprises increased activity at elevated temperatures as compared to a wildtype or reference transposase.In some further embodiments, the engineered transposases comprise increased adapter loading or adapter ligation (i.e. tagging) as compared to a wild-type or reference transposase.In some additional embodiments, the engineered transposases comprise increased binding to a target polynucleotide, as compared to a wild-type or reference transposase.In yet some further embodiments, the engineered transposases comprise decreased sequence insertion bias, as compared to a wild-type or reference transposase.In some additional embodiments, the engineered polypeptide comprises an amino acid sequence with at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or greater sequence identity to any polypeptide sequence set forth in SEQ ID NOs: 2-2368, 2387-5694, and 5706-5756.In some additional embodiments, the engineered transposase comprises a polypeptide sequence selected from the polypeptide sequences between SEQ ID NOs: 2-2368, 2388-5694, and 5706-5756.In yet some additional embodiments, the present invention provides purified engineered transposases.The present invention also provides compositions comprising at least one engineered transposase provided herein.The present invention also provides engineered polynucleotide sequences encoding at least one engineered transposase provided herein.In some embodiments, the present invention provides engineered polynucleotide sequences comprising at least 85%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to the reference sequence of SEQ ID NOs: 1 , 3, 155, 301 , 337, 605, 971 , 1081 , 1217, 1579, 1597, 1599, 1603, 1719, 2367, 2425, 2539, 3401 , 3661 , 3787, 4149, 4277, 4641 , 4983, 5195, 5285, 5411 , 5421 , and / or 5703-5755, and / or or a functional fragment thereof, wherein said polynucleotide sequence encodes an engineered polypeptide comprising at least one substitution at one or more amino acid positions. In some embodiments, a functional fragment of an engineered polynucleotide sequence does not contain the segment of nucleotides encoding the C-terminal peptide spacer GGSG (SEQ ID NO: 5777) and / or the 6-hisitine tag.In some further embodiments, the engineered polynucleotide sequences encode at least one engineered transposase comprising a sequence having at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to the reference sequence of SEQ ID NOs: 2, 4, 156, 302, 338, 606, 972, 1082, 1218, 1580, 1598, 1600, 1604, 1720, 2368, 2426, 2540, 3402, 3662, 3788, 4150, 4278, 4642, 4984, 5196, 5286, 5412, 5422, and / or 5704.In some additional embodiments, the engineered polynucleotide sequence comprises SEQ ID NOs: 1 , 3, 155, 301 , 337, 605, 971 , 1081 , 1217, 1579, 1597, 1599, 1603, 1719, 2367, 2425, 2539, 3401 , 3661 , 3787, 4149, 4277, 4641 , 4983, 5195, 5285, 5411 , 5421 , and / or 5705-5755.In yet some additional embodiments, the engineered polynucleotide sequence is operably linked to a control sequence.In still some other embodiments, the engineered polynucleotide sequence is codon-optimized.The present invention also provides an engineered polynucleotide encoding at least one engineered polypeptide described in the above paragraphs. In some embodiments, the engineered polynucleotide comprises the polynucleotide sequences set forth in SEQ ID NOs: 1 -2367, SEQ ID NOs: 2385-5693, and SEQ ID NOs: 5705-5755. In some embodiments, the engineered polynucleotide comprises the polynucleotide sequences set forth in SEQ ID NO: 2395. In some embodiments, the engineered polynucleotide comprises the polynucleotide sequences set forth in SEQ ID NO: 4641 . In some embodiments, the engineered polynucleotide comprises the polynucleotide sequences set forth in SEQ ID NO: 4871 . In some embodiments, the engineered polynucleotide comprises the polynucleotide sequences set forth in SEQ ID NO: 4953. In some embodiments, the engineered polynucleotide comprises the polynucleotide sequences set forth in SEQ ID NO: 5185. In some embodiments, the engineered polynucleotide comprises the polynucleotide sequences set forth in SEQ ID NO: 5353. In some embodiments, the engineered polynucleotide comprises the polynucleotide sequences set forth in SEQ ID NO: 5499. In some embodiments, the engineered polynucleotide comprises the polynucleotide sequences set forth in SEQ ID NO: 5519. In some embodiments, the engineered polynucleotide comprises the polynucleotide sequences set forth in SEQ ID NO: 5701 . In some embodiments, the engineered polynucleotide comprises the polynucleotide sequences set forth in SEQ ID NO: 5737. In some embodiments, the engineered polynucleotide comprises one of the foregoing polynucleotide sequence or a polynucleotide sequence encoding an engineered polypeptide described herein (e.g., an engineered transposase), wherein the segment of nucleotides encoding the C-terminal peptide spacer GGSG (SEQ ID NO: 5777) and / or the 6-histidine tag is modified or removed. In some embodiments, the segment of nucleotides encoding the C-terminal peptide spacer GGSG (SEQ ID NO: 5777) is modified or replaced to encode a different C-terminal peptide spacer. In some embodiments, the segment of nucleotides encoding the C-terminal 6-histidine tag is modified or replaced to encode a different tag. In some embodiments, the segment of nucleotides encoding the C-terminal peptide spacer GGSG (SEQ ID NO: 5777) and / or the 6-histidine tag is removed.The present invention further provides vectors comprising at least one engineered polynucleotide described above. In some embodiments, the vectors further comprise at least one control sequence.The present invention also provides host cells comprising the vectors provided herein. In some embodiments, the host cell produces at least one engineered polypeptide provided herein.The present invention further provides methods of producing an engineered transposase polypeptide, comprising the steps of culturing the host cell provided herein under conditions such that the engineered polynucleotide is expressed and the engineered polypeptide is produced. In some embodiments, the methods further comprise the step of recovering the engineered polypeptide. In some embodiments, the methods further comprise the step of purifying at least one engineered transposase.In some embodiments, the invention provides, a method for generating a mixture of altered nucleic acids comprising exposing a target nucleic acid to an engineered transposase under conditions and for a time sufficient for the engineered transposase to carry out a transposition event, wherein the engineered transposase comprises a polypeptide sequence comprising at least 85% sequence identity to a reference sequence of SEQ ID NOs: 2, 4, 156, 302, 338, 606, 972, 1082, 1218, 1580, 1598, 1600, 1604, 1720, 2368, 2426, 2540, 3402, 3662, 3788, 4150, 4278, 4642, 4984, 5196, 5286, 5412, 5422, and / or 5704, or a functional fragment thereof, wherein the engineered transposase comprises at least one substitution or substitution set in its polypeptide sequence relative to the reference sequence.In some embodiments, the altered nucleic acids are fragmented from the target nucleic acid.In some embodiments, the invention provides, a method of preparing a nucleic acid library comprising:(a) contacting a target nucleic acid with an engineered transposase under conditions and for a time sufficient for the engineered transposase to carry out a transposition event, wherein the engineered transposase comprises a polypeptide sequence comprising at least 85% sequence identity to a reference sequence of SEQ ID NOs: 2, 4, 156, 302, 338, 606, 972, 1082, 1218, 1580, 1598, 1600, 1604, 1720, 2368, 2426, 2540, 3402, 3662, 3788, 4150, 4278, 4642, 4984, 5196, 5286, 5412, 5422, and / or 5704, or a functional fragment thereof, wherein the engineered transposase comprises at least one substitution or substitution set in its polypeptide sequence relative to the reference sequences;(b) fragmenting the target nucleic acid into a plurality of nucleic acid fragments;(c) selecting a subset of nucleic acid fragments from the plurality of nucleic acid fragments; and(d) amplifying the subset of nucleic acid fragments to generate the nucleic acid library.In some embodiments, the invention provides, a method of sequencing a target nucleic acid comprising:(a) contacting the target nucleic acid with an engineered transposase under conditions and for a time sufficient for the engineered transposase to carry out a transposition event, wherein the engineered transposase comprises a polypeptide sequence comprising at least 85% sequence identity to a reference sequence of SEQ ID NOs: 2, 4, 156, 302, 338, 606, 972, 1082, 1218, 1580, 1598, 1600, 1604, 1720, 2368, 2426, 2540, 3402, 3662, 3788, 4150, 4278, 4642, 4984, 5196, 5286, 5412, 5422, and / or 5704, or a functional fragment thereof, wherein the engineered transposase comprises at least one substitution or substitution set in its polypeptide sequence relative to the reference sequence;(b) fragmenting the target nucleic acid into a plurality of nucleic acid fragments;(c) selecting a subset of nucleic acid fragments from the plurality of nucleic acid fragments;(d) amplifying the subset of nucleic acid fragments to generate a nucleic acid library; and(e) sequencing the nucleic acid library.In some embodiments, the method further includes adding a polynucleotide to the 5’ and / or 3’ end of each nucleic acid fragment of the plurality of nucleic acid fragments.In some embodiments, the engineered transposase is any one of the engineered transposases of the foregoing embodiments. In some embodiments, the engineered transposase comprises thepolypeptide sequence of SEQ ID NO: 2396. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 4642. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 4872. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 4954. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 5186. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 5354. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 5500. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 5520. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 5702. In some embodiments, the engineered transposase comprises the polypeptide sequence of SEQ ID NO: 5738. In some embodiments, the polypeptide sequence of the engineered transposase is modified to replace or remove the C-terminal peptide spacer GGSG (SEQ ID NO: 5777) and / or the 6-histidine tag. In some embodiments, the polypeptide sequence of the engineered transposase is modified to replace the C-terminal peptide spacer GGSG (SEQ ID NO: 5777) with a different peptide spacer. In some embodiments, the polypeptide sequence of the engineered transposase is modified to replace the 6-histidine tag with a different tag. In some embodiments, the polypeptide sequence of the engineered transposase is modified to remove the C-terminal histidine tag and the peptide spacer GGSG (SEQ ID NO: 5777) or a portion thereof.In some embodiments, the engineered transposase is purified.In some embodiments, fragmenting the target nucleic acid comprises tagmentation or random sheering and adapter ligation. In some embodiments, fragmenting the target nucleic acid comprises tagmentation.In some embodiments, each altered nucleic acid of the mixture of altered nucleic acids comprises an identifiable sequence tag (1ST).In some embodiments, each nucleic acid fragment of the subset of nucleic acid fragments comprises an identifiable sequence tag (1ST). In some embodiments, the 1ST is between 6 and 30 nucleotides in length (e.g., 6-10, 10-15, 15-20, 20-25, 25-30, such as 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides in length).In some embodiments, each altered nucleic acid of the mixture of altered nucleic acids is between 30 and 30000 nucleotides in length (e.g., 30-30000, 30-25000, 30-20000, 30-15000, 30-10000, 30-7500, 30-5000, 30-4000, 30-3000, 30-2500, 30-2000, 30-1500, 30-1000, 30-900, 30-800, 30-700, 30-600, 30- 500, 30-400, 30-300, 30-250, 30-200, 30-150, 30-100, 30-50, 100-500, 100-1000, 100-1500, 100-2500, 100-5000, 100-10000, 100-20000, 100-30000, 500-1000, 500-2500, 500-5000, 500-10000, 500-20000, 500-30000, 1000-2500, 1000-5000, 1000-10000, 1000-20000, 1000-30000, 5000-10000, 5000-20000, 5000-30000, 10000-20000, 20000-30000, 2500-5000, 7500-10000, or 15000-25000 nucleotides in length).In some embodiments, each nucleic acid fragment of the plurality of nucleic acid fragments is between 30 and 30000 nucleotides in length (e.g., 30-30000, 30-25000, 30-20000, 30-15000, 30-10000, 30-7500, 30-5000, 30-4000, 30-3000, 30-2500, 30-2000, 30-1500, 30-1000, 30-900, 30-800, 30-700, 30- 600, 30-500, 30-400, 30-300, 30-250, 30-200, 30-150, 30-100, 30-50, 100-500, 100-1000, 100-1500, 100-2500, 100-5000, 100-10000, 100-20000, 100-30000, 500-1000, 500-2500, 500-5000, 500-10000, 500- 20000, 500-30000, 1000-2500, 1000-5000, 1000-10000, 1000-20000, 1000-30000, 5000-10000, 5000- 20000, 5000-30000, 10000-20000, 20000-30000, 2500-5000, 7500-10000, or 15000-25000 nucleotides in length).In some embodiments, the transposition event occurs in reaction conditions additionally comprising a terminal deoxynucleotide transferase, dNTP, and / or buffer components suitable for the addition of deoxynucleotides to the 3’ terminus of each of the plurality of nucleic acid fragments.In some embodiments, the transposition event occurs in reaction conditions additionally comprising a DNA ligase and / or buffer components suitable for a ligation reaction.In some embodiments, each altered nucleic acid of the mixture of altered nucleic acids further comprises a sample tag and / or a unique molecular identifier (UMI). In some embodiments, each nucleic acid fragment of the subset of nucleic acid fragments further comprises a sample tag and / or a unique molecular identifier (UMI).In some embodiments, amplification of the subset of nucleic acid fragments is performed through polymerase chain reaction (PCR), multiple displacement amplification (MDA), ligase chain reaction (LCR), loop mediated isothermal amplification (LAMP), rolling circle amplification (RCA), or strand displacement amplification (SDA).In some embodiments, the sequencing comprises next-generation sequencing (NGS).In some embodiments, the sequencing comprises sequencing by synthesis, sequencing by ligation, or nanopore sequencing. In some embodiments, the sequencing by synthesis comprises Illumina™ dye sequencing, single-molecule real-time (SMRT™) sequencing, or pyrosequencing. In some embodiments, the sequencing by ligation comprises polony-based sequencing or SOLiD™ sequencing.In some embodiments, the target nucleic acid comprises genomic DNA or cDNAs from a single cell. In some embodiments, the target nucleic acid comprises nucleic acids from a plurality of haplotypes.In some embodiments, the target nucleic acid is crosslinked via histones or chromatin from single or multiple cells. In some embodiments, the target nucleic acid has been condensed or optionally treated with one or more condensing agents.BRIEF DESCRIPTION OF THE DRAWINGSFigure 1 depicts active transposase variants, bound to appropriate adapter sequences, that are capable of binding target sequences, cleaving the target strand, and ligating the adapter oligonucleotides to the target fragment.Figure 2 provides a scheme depicting the capillary electrophoresis (CE) assay used in the Examples to assess the activity of the engineered transposase variants.Figure 3 is a plot of fragment sizes of NGS libraries prepared from 50 ng input DNA sample.Libraries prepared with a transposase that has the polypeptide sequence set forth in SEQ ID NO: 5704 or a commercially available Tn5 transposase are indicated.Figures 4A through 4D are plots showing the insertion bias of the indicated transposases as determined by the calculated root mean square error (RMSE) of the first 25 bases of the insert.Figures 5A and 5B are gels showing the results of the enzyme activity assay comparing engineered transposase (SequenceK; Figure 5A) with a commercial transposase (hyperactive Tn5) for their ability to convert circular plasmid to linear form. 2.5 pmol transposase was used in the comparison. Plasmids are present in the gel as the higher length bars, i.e. >3000 bp, while the linear form fragments are shown as the smear < 3000 bp.Figure 6 is a plot of fragment sizes of NGS libraries prepared from 50 ng - 200 ng input DNA sample.Figure 7 is a plot showing the estimated NGS library size generated from 100 ng of human NA12878 genomic DNA using different engineered transposases (SequenceE, Sequencer, SequenceG, SequenceH, SequenceK, and SequenceL) and commercial hyperactive Tn5 transposases (Hyp-Tn5).Figure 8 is a plot showing GC coverage bias of NGS libraries generated from 100 ng of human NA12878 genomic DNA using different engineered transposases (SequenceE; SequenceK; SequenceF; SequenceG; SequenceH; and SequenceL) and commercial hyperactive Tn5 transposases.Figure 9 is a plot showing fold-coverage of NGS libraries generated from 100 ng of human NA12878 genomic DNA using different engineered transposases (SequenceE; SequenceK; SequenceF; SequenceG; SequenceH; and SequenceL) and commercial hyperactive Tn5 transposases.Figures 10A through 10D are plots showing the per base sequence content of the transposition sites of the different commercial transposases and engineered transposases. Each nucleotide (A, T, G, or C) is represented by a single line trace, representing the percentage of reads having that nucleotide at the corresponding position relative to the read start site (transposition site) of the DNA molecules in the sequenced library.Figure 11 are plots showing GC coverage bias of NGS libraries prepared by ExpressPlex using hyperactive Tn5 transposase and the SequenceH engineered transposase from Staphylococcus epidermidis, Bacillus subtilis, Escherichia coll, and Pseudomonas aeruginosa.Figure 12 is a plot showing the coverage profiles of NGS libraries prepared by ExpressPlex of lambda amplicon 18 DNA samples using a commercial and engineered (SequenceE, SequenceH, and SequenceK) transposases. Data is shown as the raw coverage profile as well as the summary statistic in the form of the coefficient of variation (CV) of the raw coverage profile.Figure 13 is a plot showing the coverage profiles of NGS libraries prepared by ExpressPlex of pUC19 DNA samples using commercial and engineered (SequenceE, SequenceH, and SequenceK) transposases. Data is shown as the raw coverage profile as well as the summary statistic in the form of the coefficient of variation (CV) of the raw coverage profile.DESCRIPTION OF THE INVENTIONUnless defined otherwise, all technical and scientific terms used herein generally have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Generally, the nomenclature used herein and the laboratory procedures of cell culture, molecular genetics, microbiology, organic chemistry, analytical chemistry and nucleic acid chemistry described below are those well-known and commonly employed in the art. Such techniques are well-known and described in numerous texts and reference works well known to those of skill in the art. Standardtechniques, or modifications thereof, are used for chemical syntheses and chemical analyses. All patents, patent applications, articles and publications mentioned herein, both supra and infra, are hereby expressly incorporated herein by reference.Although any suitable methods and materials similar or equivalent to those described herein find use in the practice of the present invention, some methods and materials are described herein. It is to be understood that this invention is not limited to the particular methodology, protocols, and reagents described, as these may vary, depending upon the context they are used by those of skill in the art. Accordingly, the terms defined immediately below are more fully described by reference to the invention as a whole.It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the present invention. The section headings used herein are for organizational purposes only and not to be construed as limiting the subject matter described. Numeric ranges are inclusive of the numbers defining the range. Thus, every numerical range disclosed herein is intended to encompass every narrower numerical range that falls within such broader numerical range, as if such narrower numerical ranges were all expressly written herein. It is also intended that every maximum (or minimum) numerical limitation disclosed herein includes every lower (or higher) numerical limitation, as if such lower (or higher) numerical limitations were expressly written herein.As used in this specification and the appended claims, the singular forms “a”, “an” and “the” include plural referents unless the context clearly indicates otherwise. Thus, for example, reference to “a polypeptide” includes more than one polypeptide. Similarly, “comprise,” “comprises,” “comprising” “include,” “includes,” and “including” are interchangeable and not intended to be limiting.It is to be understood that where descriptions of various embodiments use the term “comprising,” those skilled in the art would understand that in some specific instances, an embodiment can be alternatively described using language “consisting essentially of” or “consisting of.” It is to be further understood that where descriptions of various embodiments use the term “optional” or “optionally” the subsequently described event or circumstance may or may not occur, and that the description includes instances where the event or circumstance occurs and instances in which it does not. It is to be understood that both the foregoing general description, and the following detailed description are exemplary and explanatory only and are not restrictive of this disclosure. The section headings used herein are for organizational purposes only and not to be construed as limiting the subject matter described.Abbreviations:The abbreviations used for the genetically encoded amino acids are conventional and are as follows:When the three-letter abbreviations are used, unless specifically preceded by an “L” or a “D” or clear from the context in which the abbreviation is used, the amino acid may be in either the L- or D- configuration about a-carbon (Ca). For example, whereas “Ala” designates alanine without specifying the configuration about the a-carbon, “D-Ala” and “L-Ala” designate D-alanine and L-alanine, respectively.When the one-letter abbreviations are used, upper case letters designate amino acids in the L- configuration about the a-carbon and lower case letters designate amino acids in the D-configuration about the a-carbon. For example, “A” designates L-alanine and “a” designates D-alanine. When polypeptide sequences are presented as a string of one-letter or three-letter abbreviations (or mixtures thereof), the sequences are presented in the amino (N) to carboxy (C) direction in accordance with common convention.The abbreviations used for the genetically encoding nucleosides are conventional and are as follows: adenosine (A); guanosine (G); cytidine (C); thymidine (T); and uridine (U). These abbreviations are also used interchangeably for nucleosides and nucleotides (nucleosides with one or more phosphate groups). Unless specifically delineated, the abbreviated nucleosides or nucleotides may be either ribonucleosides (or ribonucleotides) or 2’-deoxyribonucleosides (or 2’-deoxyribonucleotides). The nucleosides or nucleotides may also be modified at the 3’ position. The nucleosides or nucleotides may be specified as being either ribonucleosides (or ribonucleotides) or 2’-deoxyribonucleosides (or 2’- deoxyribonucleotides) on an individual basis or on an aggregate basis. When nucleic acid sequences arepresented as a string of one-letter abbreviations, the sequences are presented in the 5’ to 3’ direction in accordance with common convention, and the phosphates are not indicated.Definitions:In reference to the present invention, the technical and scientific terms used in the descriptions herein will have the meanings commonly understood by one of ordinary skill in the art, unless specifically defined otherwise. Accordingly, the following terms are intended to have the following meanings.“EC” number refers to the Enzyme Nomenclature of the Nomenclature Committee of the International Union of Biochemistry and Molecular Biology (NC-IUBMB). The IUBMB biochemical classification is a numerical classification system for enzymes based on the chemical reactions they catalyze.“ATCC” refers to the American Type Culture Collection whose biorepository collection includes genes and strains.“NCBI” refers to National Center for Biological Information and the sequence databases provided therein.“Protein,” “polypeptide,” and “peptide” are used interchangeably herein to denote a polymer of at least two amino acids covalently linked by an amide bond, regardless of length or post-translational modification (e.g., glycosylation, phosphorylation, lipidation, myristilation, ubiquitination, etc.). Included within this definition are D- and L-amino acids, and mixtures of D- and L-amino acids, as well as polymers comprising D- and L-amino acids, and mixtures of D- and L-amino acids.“Amino acids” are referred to herein by either their commonly known three-letter symbols or by the one-letter symbols recommended by IUPAC-IUB Biochemical Nomenclature Commission. Nucleotides, likewise, may be referred to by their commonly accepted single letter codes.As used herein, “polynucleotide,” “oligonucleotide,” and “nucleic acid” are used interchangeably herein and refer to two or more nucleosides or nucleotides that are covalently linked together. The polynucleotide may be wholly comprised of ribonucleotides (i.e., RNA), wholly comprised of 2’ deoxyribonucleotides (i.e., DNA), wholly comprised of other synthetic nucleotides or comprised of mixtures of synthetic, ribo- and / or 2’ deoxyribonucleotides. The polynucleotides may also include modified nucleotides with substitutions, including 2’ substitutions (e.g., 2’-fluoro, 2’-O-methyl, 2’-O-methoxyethyl, locked or constrained ethyl modifications, and others known to those skilled in the art). Nucleosides will be linked together via standard phosphodiester linkages or via one or more non-standard linkages, including but not limited to phosphorothioate linkages. The polynucleotide may be single-stranded or doublestranded or may include both single-stranded regions and double-stranded regions. Moreover, while a polynucleotide will typically be composed of the naturally occurring encoding nucleobases (i.e., adenine, guanine, uracil, thymine and cytosine), it may include one or more modified and / or synthetic nucleobases, such as, for example, inosine, xanthine, hypoxanthine, etc. In some embodiments, such modified or synthetic nucleobases are nucleobases encoding amino-acid sequences. Nucleobases that are modified or synthetic may comprise any known or hypothetical or future discovered modification or structure that would be recognized by one of skill in the art as a modified or synthetic nucleobase. Examples of modified nucleobases or nucleotides include, but are not limited to, diaminopurine, 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil, hypoxanthine, xantine, 4-acetylcytosine, 5-(carboxyhydroxylmethyl)uracil, 5- carboxymethylaminomethyl-2-thiouridine, 5-carboxymethylaminomethyluracil, dihydrouracil, beta-D- galactosylqueosine, inosine, N6-isopentenyladenine, 1 -methylguanine, 1 -methylinosine, 2,2- dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-adenine, 7- methylguanine, 5-methylaminomethyluracil, 5-methoxyaminomethyl-2-thiouracil, beta-D- mannosylqueosine, 5'-methoxycarboxymethyluracil, 5-methoxyuracil, 2-methylthio-N6- isopentenyladenine, uracil-5-oxyacetic acid (v), wybutoxosine, pseudouracil, queosine, 2-thiocytosine, 5- methyl-2-thiouracil, 2-thiouracil, 4-thiouraci 1 , 5-methyluracil, uracil-5-oxyacetic acid methylester, 5-methyl- 2-thiouracil, and 3-(3-amino-3-N-2-carboxypropyl) uracil. A polynucleotide may contain one or more chemical modifications, including modifications on the 5’ end, the 3’ end, or internally. Examples of chemical modifications include, but are not limited to, addition of functional groups (e.g., biotins, amino modifiers, alkynes, thiol modifiers, or azides), fluorophores (e.g. quantum dots or organic dyes), spacers (e.g. C3 spacer, dSpacer, photo-cleavable spacers), modified bases, or modified backbones.Similarly, the terms “polynucleotide,” “oligonucleotide,” and “nucleic acid” are intended to comprise any modified or synthetic structure that is now known or discovered in the future that would be recognized by one of skill in the art as being or having the function of a “polynucleotide,” “oligonucleotide,” or “nucleic acid.” An example of a modified or synthetic structure having the function of a “polynucleotide,” “oligonucleotide,” or “nucleic acid” is PNA or peptide nucleic acid.“Coding sequence” refers to that portion of a nucleic acid (e.g., a gene) that encodes an amino acid sequence of a protein.“Naturally-occurring” or “wild-type” refers to the form found in nature. For example, a naturally occurring or wild-type polypeptide or polynucleotide sequence is a sequence present in an organism that can be isolated from a source in nature and which has not been intentionally modified by human manipulation.As used herein, “recombinant,” “engineered,” and “non-naturally occurring” when used with reference to a cell, nucleic acid, or polypeptide, refer to a material, or a material corresponding to the natural or native form of the material, that has been modified in a manner that would not otherwise exist in nature. In some embodiments, the cell, nucleic acid or polypeptide is identical to a naturally occurring cell, nucleic acid or polypeptide, but is produced or derived from synthetic materials and / or by manipulation using recombinant techniques. Non-limiting examples include, among others, recombinant cells expressing genes that are not found within the native (non-recombinant) form of the cell or expressed native genes that are otherwise expressed at a different level.“Percentage of sequence identity” and “percentage homology” are used interchangeably herein to refer to comparisons among polynucleotides or polypeptides, and are determined by comparing two optimally aligned sequences over a comparison window, wherein the portion of the polynucleotide or polypeptide sequence in the comparison window may comprise additions or deletions (i.e., gaps) as compared to the reference sequence for optimal alignment of the two sequences. The percentage may be calculated by determining the number of positions at which the identical nucleic acid base or amino acid residue occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the window of comparison and multiplying the resultby 100 to yield the percentage of sequence identity. Alternatively, the percentage may be calculated by determining the number of positions at which either the identical nucleic acid base or amino acid residue occurs in both sequences or a nucleic acid base or amino acid residue is aligned with a gap to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the window of comparison and multiplying the result by 100 to yield the percentage of sequence identity. Those of skill in the art appreciate that there are many established algorithms available to align two sequences. Optimal alignment of sequences for comparison can be conducted, e.g., by the local homology algorithm of Smith and Waterman (Smith and Waterman, Adv. Appl. Math., 2:482

[1981] ), by the homology alignment algorithm of Needleman and Wunsch (Needleman and Wunsch, J. Mol. Biol., 48:443

[1970] ), by the search for similarity method of Pearson and Lipman (Pearson and Lipman, Proc. Natl. Acad. Sci. USA 85:2444

[1988] ), by computerized implementations of these algorithms (e.g., GAP, BESTFIT, FASTA, and TFASTA in the GCG Wisconsin Software Package), or by visual inspection, as known in the art. Examples of algorithms that are suitable for determining percent sequence identity and sequence similarity include, but are not limited to the BLAST and BLAST 2.0 algorithms, which are described by Altschul et al. (See, Altschul et al., J. Mol. Biol., 215: 403-410

[1990] ; and Altschul et al., Nucl. Acids Res., 3389-3402

[1977] , respectively). Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information website. This algorithm involves first identifying high scoring sequence pairs (HSPs) by identifying short words of length W in the query sequence, which either match or satisfy some positive-valued threshold score T when aligned with a word of the same length in a database sequence. T is referred to as, the neighborhood word score threshold (Altschul et al., supra). These initial neighborhood word hits act as seeds for initiating searches to find longer HSPs containing them. The word hits are then extended in both directions along each sequence for as far as the cumulative alignment score can be increased. Cumulative scores are calculated using, for nucleotide sequences, the parameters M (reward score for a pair of matching residues; always >0) and N (penalty score for mismatching residues; always <0). For amino acid sequences, a scoring matrix is used to calculate the cumulative score. Extension of the word hits in each direction are halted when: the cumulative alignment score falls off by the quantity X from its maximum achieved value; the cumulative score goes to zero or below, due to the accumulation of one or more negative-scoring residue alignments; or the end of either sequence is reached. The BLAST algorithm parameters W, T, and X determine the sensitivity and speed of the alignment. The BLASTN program (for nucleotide sequences) uses as defaults a wordlength (W) of 11 , an expectation (E) of 10, M=5, N=-4, and a comparison of both strands. For amino acid sequences, the BLASTP program uses as defaults a wordlength (W) of 3, an expectation (E) of 10, and the BLOSUM62 scoring matrix (See, Henikoff and Henikoff, Proc. Natl. Acad. Sci. USA 89:10915

[1989] ). Exemplary determination of sequence alignment and % sequence identity can employ the BESTFIT or GAP programs in the GCG Wisconsin Software package (Accelrys, Madison Wl), using default parameters provided.“Reference sequence” refers to a defined sequence used as a basis for a sequence comparison. A reference sequence may be a subset of a larger sequence, for example, a segment of a full-length gene or polypeptide sequence. Generally, a reference sequence is at least 20 nucleotide or amino acid residues in length, at least 25 residues in length, at least 50 residues in length, or the full length of thenucleic acid or polypeptide. Since two polynucleotides or polypeptides may each (1 ) comprise a sequence {i.e., a portion of the complete sequence) that is similar between the two sequences, and (2) may further comprise a sequence that is divergent between the two sequences, sequence comparisons between two (or more) polynucleotides or polypeptide are typically performed by comparing sequences of the two polynucleotides or polypeptides over a “comparison window” to identify and compare local regions of sequence similarity. In some embodiments, a “reference sequence” can be based on a primary amino acid sequence, where the reference sequence is a sequence that can have one or more changes in the primary sequence. For instance, a “reference sequence based on SEQ ID NO:4 having at the residue corresponding to X14 a valine” or X14V refers to a reference sequence in which the corresponding residue at X14 in SEQ ID NO:4, which is a tyrosine, has been changed to valine.“Comparison window” refers to a conceptual segment of at least about 20 contiguous nucleotide positions or amino acids residues wherein a sequence may be compared to a reference sequence of at least 20 contiguous nucleotides or amino acids and wherein the portion of the sequence in the comparison window may comprise additions or deletions {i.e., gaps) of 20 percent or less as compared to the reference sequence (which does not comprise additions or deletions) for optimal alignment of the two sequences. The comparison window can be longer than 20 contiguous residues, and includes, optionally 30, 40, 50, 100, or longer windows.As used herein, “substantial identity” refers to a polynucleotide or polypeptide sequence that has at least 80 percent sequence identity, at least 85 percent identity, at least between 89 to 95 percent sequence identity, or more usually, at least 99 percent sequence identity as compared to a reference sequence over a comparison window of at least 20 residue positions, frequently over a window of at least 30-50 residues, wherein the percentage of sequence identity is calculated by comparing the reference sequence to a sequence that includes deletions or additions which total 20% or less of the reference sequence over the window of comparison. In some specific embodiments applied to polypeptides, the term “substantial identity” means that two polypeptide sequences, when optimally aligned, such as by the programs GAP or BESTFIT using default gap weights, share at least 80% sequence identity, preferably at least 89% sequence identity, at least 95% sequence identity or more (e.g., 99% sequence identity). In some embodiments, residue positions that are not identical in sequences being compared differ by conservative amino acid substitutions.“Corresponding to,” “reference to,” and “relative to” when used in the context of the numbering of a given amino acid or polynucleotide sequence refer to the numbering of the residues of a specified reference sequence when the given amino acid or polynucleotide sequence is compared to the reference sequence. In other words, the residue number or residue position of a given polymer is designated with respect to the reference sequence rather than by the actual numerical position of the residue within the given amino acid or polynucleotide sequence. For example, a given amino acid sequence, such as that of an engineered transposase, can be aligned to a reference sequence by introducing gaps to optimize residue matches between the two sequences. In these cases, although the gaps are present, the numbering of the residue in the given amino acid or polynucleotide sequence is made with respect to the reference sequence to which it has been aligned.“Amino acid difference” or “residue difference” refers to a change in the amino acid residue at a position of a polypeptide sequence relative to the amino acid residue at a corresponding position in a reference sequence. The positions of amino acid differences generally are referred to herein as “Xn,” where n refers to the corresponding position in the reference sequence upon which the residue difference is based. For example, a “residue difference at position X25 as compared to SEQ ID NO: 2” refers to a change of the amino acid residue at the polypeptide position corresponding to position 25 of SEQ ID NO:2. Thus, if the reference polypeptide of SEQ ID NO: 2 has a valine at position 25, then a “residue difference at position X25 as compared to SEQ ID NO:2” an amino acid substitution of any residue other than valine at the position of the polypeptide corresponding to position 25 of SEQ ID NO: 2. In most instances herein, the specific amino acid residue difference at a position is indicated as “XnY” where “Xn” specified the corresponding position as described above, and “Y” is the single letter identifier of the amino acid found in the engineered polypeptide (i.e., the different residue than in the reference polypeptide). In some embodiments, more than one amino acid can appear in a specified residue position (i.e., the alternative amino acids can be listed in the form XnY / Z, where Y and Z represent alternate amino acid residues). In some instances (e.g., in Tables 5.2, 6.2, 7.2, 8.2, 9.2, 10.2, 10.3, 10.5, 11 .2, 11 .4, 11 .5, 12.2, 12.3, 12.5, 12.6, 13.2, 13.3, 13.4, 14.2, 14. 3, 14.5, 14.6, 14.8, 14.9, 15.1 , 16.2, 16.3, 17.2, 17.3, 18.2,19.2, 19.3, 20.2, 20.3, 21 .2, 21 .3, 22.2, 23.2, 24.2, 25.2, 26.2, 27.2, 28.2, 29.2, 30.2, 31 .2, 32.2, 33.2,34.2, 35.1 , 36.1 , and 36.3) the present invention also provides specific amino acid differences denoted by the conventional notation “AnB”, where A is the single letter identifier of the residue in the reference sequence, “n” is the number of the residue position in the reference sequence, and B is the single letter identifier of the residue substitution in the sequence of the engineered polypeptide. Furthermore, in some instances, a polypeptide of the present invention can include one or more amino acid residue differences relative to a reference sequence, which is indicated by a list of the specified positions where changes are made relative to the reference sequence. In some additional embodiments, the present invention provides engineered polypeptide sequences comprising both conservative and non-conservative amino acid substitutions.As used herein, “conservative amino acid substitution” refers to a substitution of a residue with a different residue having a similar side chain, and thus typically involves substitution of the amino acid in the polypeptide with amino acids within the same or similar defined class of amino acids. By way of example and not limitation, an amino acid with an aliphatic side chain is substituted with another aliphatic amino acid (e.g., alanine, valine, leucine, and isoleucine); an amino acid with an hydroxyl side chain is substituted with another amino acid with a hydroxyl side chain (e.g., serine and threonine); an amino acid having an aromatic side chain is substituted with another amino acid having an aromatic side chain (e.g., phenylalanine, tyrosine, tryptophan, and histidine); an amino acid with a basic side chain is substituted with another amino acid with a basic side chain (e.g., lysine and arginine); an amino acid with an acidic side chain is substituted with another amino acid with an acidic side chain (e.g., aspartic acid or glutamic acid); and / or a hydrophobic or hydrophilic amino acid is replaced with another hydrophobic or hydrophilic amino acid, respectively. Exemplary conservative substitutions are provided in Table 1.“Non-conservative substitution” refers to substitution of an amino acid in the polypeptide with an amino acid with significantly differing side chain properties. Non-conservative substitutions may use amino acids between, rather than within, the defined groups and affects (a) the structure of the peptide backbone in the area of the substitution (e.g., proline for glycine), (b) the charge or hydrophobicity, or (c) the bulk of the side chain. By way of example and not limitation, an exemplary non-conservative substitution can be an acidic amino acid substituted with a basic or aliphatic amino acid; an aromatic amino acid substituted with a small amino acid; and a hydrophilic amino acid substituted with a hydrophobic amino acid.“Deletion” refers to modification to the polypeptide by removal of one or more amino acids from the reference polypeptide. Deletions can comprise removal of 1 or more amino acids, 2 or more amino acids, 5 or more amino acids, 10 or more amino acids, 15 or more amino acids, or 20 or more amino acids, up to 10% of the total number of amino acids, or up to 20% of the total number of amino acids making up the reference enzyme while retaining enzymatic activity and / or retaining the improved properties of an engineered transposase enzyme. Deletions can be directed to the internal portions and / or terminal portions of the polypeptide. In various embodiments, the deletion can comprise a continuous segment or can be discontinuous.“Insertion” refers to modification to the polypeptide by addition of one or more amino acids from the reference polypeptide. In some embodiments, the improved engineered transposase enzymes comprise insertions of one or more amino acids to the naturally occurring polypeptide as well as insertions of one or more amino acids to other improved transposase polypeptides. Insertions can be in the internal portions of the polypeptide, or to the carboxy or amino terminus. Insertions as used herein include fusion proteins as is known in the art. The insertion can be a contiguous segment of amino acids or separated by one or more of the amino acids in the naturally occurring polypeptide.“Fragment” as used herein refers to a polypeptide that has an amino-terminal and / or carboxyterminal deletion, but where the remaining amino acid sequence is identical to the corresponding positions in the sequence. Fragments can be at least 14 amino acids long, at least 20 amino acids long, at least 50 amino acids long or longer, and up to 70%, 80%, 90%, 95%, 98%, and 99% of the full-length transposase polypeptide, for example the polypeptide of SEQ ID NO: 2 or a transposase provided in the polypeptide sequences of SEQ ID NOs: 4-2368, 2388-5694, and 5706-5756.“Isolated polypeptide” refers to a polypeptide which is substantially separated from other contaminants that naturally accompany it, e.g., protein, lipids, and polynucleotides. The term embraces polypeptides which have been removed or purified from their naturally-occurring environment or expression system {e.g., host cell or in vitro synthesis). The engineered transposase enzymes may be present within a cell, present in the cellular medium, or prepared in various forms, such as lysates or isolated preparations. As such, in some embodiments, the engineered transposase enzyme can be an isolated polypeptide.“Substantially pure polypeptide” refers to a composition in which the polypeptide species is the predominant species present ( / .e., on a molar or weight basis it is more abundant than any other individual macromolecular species in the composition) and is generally a substantially purified composition when the object species comprises at least about 50 percent of the macromolecular species present by mole or % weight. Generally, a substantially pure transposase composition will comprise about 60 % or more, about 70% or more, about 80% or more, about 90% or more, about 95% or more, and about 98% or more of all macromolecular species by mole or % weight present in the composition. In some embodiments, the object species is purified to essential homogeneity ( / .e., contaminant species cannot be detected in the composition by conventional detection methods) wherein the composition consists essentially of a single macromolecular species. Solvent species, small molecules (<500 Daltons), and elemental ion species are not considered macromolecular species. In some embodiments, the isolated engineered transposase polypeptide is a substantially pure polypeptide composition.As used herein, a “target nucleic acid” refers to any nucleic acid (e.g., RNA or DNA) of interest that is selected for amplification or analysis (e.g., sequencing) using a composition (e.g., sequencing oligonucleotides or barcoding oligonucleotides) or method of the invention. In some instances, RNA may be converted to cDNA prior to being used in the methods described herein. In some instances, target nucleic acid includes genomic DNA or cDNAs from a single cell. In some instances, target nucleic acid includes nucleic acids from a plurality of haplotypes. In some instances, target nucleic acid is crosslinked via histones or chromatin from single or multiple cells. In some instances, target nucleic acid has been condensed or optionally treated with one or more condensing agents.As used herein, “improved enzyme property” refers to at least one improved property of an enzyme. In some embodiments, the present invention provides engineered transposase polypeptides that exhibit an improvement in any enzyme property as compared to a reference transposase polypeptide and / or a wild-type transposase polypeptide, and / or another engineered transposase polypeptide. Thus, the level of “improvement” can be determined and compared between various transposase polypeptides, including wild-type, as well as engineered transposases. Improved properties include, but are not limited, to such properties as increased polynucleotide fragmentation or cleavage activity, increased thermostability, increased activity at elevated temperatures, increased soluble expression, decreased inhibition, increased adapter loading (donor binding) or adapter ligation (i.e. tagging), increased binding to a target polynucleotide, and / or decreased sequence insertion bias as compared to another wild-type transposase or engineered transposase.“Sequence insertion bias,” “target bias,” “sequence bias,” or related terms used herein refer to the degree to which a transposome cleaves target nucleotide sequences in a non-random manner based onthe composition of the sequence of the polynucleotide. For example, a transposase with G / C target bias will have an increased rate of binding to G / C rich sequences, as compared to other sequences. As detailed in the Examples below, sequence insertion bias can be approximated or quantified using several methods. One method evaluates bias over the first 15 nucleotides of NGS reads putatively comprising the region of the target bound by the transposome construct. Bias plots generated from NGS data represent this bias as a function of the distance from the insertion site. Overall relative bias for each variant can be calculated as the root-mean-squared deviation (RMSD) of the observed vs. expected representation of all four nucleotides at each position. Conversely, “insertion promiscuity” and “promiscuity” refer to the degree to which a transposome binds to target nucleotide sequences in a random manner, without regard to the sequence composition of the polynucleotide.“Next generation sequencing” refers to modern high-throughput or massively-parallel sequencing techniques that allow sequencing of many polynucleotide sequences at the same time, as compared to traditional sequencing methods, including the Sanger method, that allowed sequencing of only one polynucleotide at a time. Next generation sequencing methods have facilitated whole genome sequencing and enabled genomics and related fields by requiring less polynucleotide sample and having greater accuracy.“Adapter,” as used herein, refers to a donor polynucleotide that contains the end sequences for transposase binding, as well as other optional application-specific elements. These may include binding sites for sequencing primers, library amplification primers, sample indexes, or other functional donor polynucleotide sequence. The adapter is loaded onto or bound to the transposase, which dimerizes to form the “transposome”.“Barcode,” as used herein, refers to a unique oligonucleotide sequence that may allow the corresponding oligonucleotide to be identified. In some instances, the nucleic acid sequence may be located at a specific position in a longer nucleic acid sequence.“Complement” or “complementary,” as used herein in reference to a sequence, refers to the sequence of a first nucleic acid in relation to that of a second nucleic acid, wherein when the first and second nucleic acids are aligned antiparallel (5’ end of the first nucleic acid matched to the 3’ end of the second nucleic acid, and vice versa) to each other, the nucleotide bases at each position in their sequences will have complementary structures following a lock-and-key principle (i.e. , A will be paired with U or T and G will be paired with C). Complementary sequences may include mismatches of up to one third of nucleotide bases. For example, two sequences that are nine bases in length may have mismatches of at most 3, at most 2, or at most 1 , or at most 0 nucleotide bases, and remain complementary to one another.“Transposition” or a “transposition event,” as used herein, is the cleavage of a target dsDNA (or hybrid DNA / RNA) sequence and ligation of the pre-loaded adapter (or another sequence containing transposon end sequences) from the transposome onto the target dsDNA at the site of cleavage. In cells, transposition typically refers to the relocation of a segment of DNA to another locus in the genome. In vitro, transposition may result in the fragmentation of a target DNA sample with the simultaneous addition of the adapter sequences to the fragmented DNA ends, i.e., tagmentation.“Synaptic complex” or “transposase synaptic complex,” as used herein, refers to a protein-nucleic acid complex including one or more transposases and one or more oligonucleotides. In some instances, the one or more oligonucleotides of the synaptic complex are inserted into a nucleic acid sequence of a nucleic acid sample by transposase activity. In some instances, the synaptic complex may include two transposases and two oligonucleotides. In some instances, the insertion of the two oligonucleotides into the nucleic acid sequence of the nucleic acid sample results in fragmentation of the nucleic acid sequence at the site of insertion. In some instances, the transposases may be Tn5 transposases. In some instances, the oligonucleotides may be adapter sequences. In some instances, the synaptic complex is pre-assembled.“Increased enzymatic activity” refers to an improved property of the transposase polypeptides, which can be represented by an increase in specific activity {e.g., product produced / time / weight protein) or an increase in percent conversion of the substrate to the product {e.g., percent conversion of starting amount of substrate to product in a specified time period using a specified amount of transposase) as compared to the reference transposase enzyme. Exemplary methods to determine enzyme activity are provided in the Examples. Any property relating to enzyme activity may be affected, including the classical enzyme properties of Km, Vmaxor kcat, changes of which can lead to increased enzymatic activity. Improvements in enzyme activity can be from about 1 .2 times the enzymatic activity of the corresponding wild-type enzyme, to as much as 2 times, 5 times, 10 times, 20 times, 25 times, 50 times or more enzymatic activity than the naturally occurring or another engineered transposase from which the transposase polypeptides were derived. Transposase activity can be measured by any one of standard assays, such as by monitoring changes in properties of substrates, cofactors, or products. In some embodiments, the amount of products generated can be measured by Liquid Chromatography-Mass Spectrometry (LC-MS), HPLC, capillary electrophoresis or other methods, as known in the art. Comparisons of enzyme activities are made using a defined preparation of enzyme, a defined assay under a set condition, and one or more defined substrates, as further described in detail herein. Generally, when lysates are compared, the numbers of cells and the amount of protein assayed are determined as well as use of identical expression systems and identical host cells to minimize variations in amount of enzyme produced by the host cells and present in the lysates.“Conversion” refers to the enzymatic conversion of the substrate(s) to the corresponding product(s). “Percent conversion” refers to the percent of the substrate that is converted to the product within a period of time under specified conditions. Thus, the “enzymatic activity” or “activity” of a transposase polypeptide can be expressed as “percent conversion” of the substrate to the product.“Thermostable” refers to a polypeptide that maintains similar activity (more than 60% to 80% for example) after exposure to elevated temperatures {e.g., 40-80 °C) for a period of time {e.g., 0.5-24 hrs) compared to the wild-type enzyme exposed to the same elevated temperature.“Solvent stable” refers to a polypeptide that maintains similar activity (more than e.g., 60% to 80%) after exposure to varying concentrations {e.g., 5-99%) of solvent (ethanol, isopropyl alcohol, dimethylsulfoxide (DMSO), tetrahydrofuran, 2-methyltetrahydrofuran, acetone, toluene, butyl acetate, methyl tert-butyl ether, etc.) for a period of time {e.g., 0.5-24 hrs) compared to the wild-type enzyme exposed to the same concentration of the same solvent.“Thermo- and solvent stable” refers to a polypeptide that is both thermostable and solvent stable. The term “stringent hybridization conditions” is used herein to refer to conditions under which nucleic acid hybrids are stable. As known to those of skill in the art, the stability of hybrids is reflected in the melting temperature (Tm) of the hybrids. In general, the stability of a hybrid is a function of ion strength, temperature, G / C content, and the presence of chaotropic agents. The Tmvalues for polynucleotides can be calculated using known methods for predicting melting temperatures (See e.g., Baldino et al., Meth. Enzymol., 168:761 -777

[1989] ; Bolton et al., Proc. Natl. Acad. Sci. USA 48:1390

[1962] ; Bresslauer et al., Proc. Natl. Acad. Sci. USA 83:8893-8897

[1986] ; Freier et al., Proc. Natl. Acad. Sci. USA 83:9373-9377

[1986] ; Kierzek et al., Biochem., 25:7840-7846

[1986] ; Rychlik et al., 1990, Nucl. Acids Res., 18:6409-6412

[1990] (erratum, Nucl. Acids Res., 19:698

[1991] ); Sambrook et al., supra); Suggs et al., 1981 , in Developmental Biology Using Purified Genes, Brown et al. [eds.], pp. 683-693, Academic Press, Cambridge, MA

[1981] ; and Wetmur, Crit. Rev. Biochem. Mol. Biol., 26:227-259

[1991] ). In some embodiments, the polynucleotide encodes the polypeptide disclosed herein and hybridizes under defined conditions, such as moderately stringent or highly stringent conditions, to the complement of a sequence encoding an engineered transposase enzyme of the present invention.“Hybridization stringency” relates to hybridization conditions, such as washing conditions, in the hybridization of nucleic acids. Generally, hybridization reactions are performed under conditions of lower stringency, followed by washes of varying but higher stringency. The term “moderately stringent hybridization” refers to conditions that permit target-DNA to bind a complementary nucleic acid that has about 60% identity, preferably about 75% identity, about 85% identity to the target DNA, with greater than about 90% identity to target-polynucleotide. Exemplary moderately stringent conditions are conditions equivalent to hybridization in 50% formamide, 5x Denhart's solution, 5xSSPE, 0.2% SDS at 42 °C, followed by washing in 0.2xSSPE, 0.2% SDS, at 42 °C. “High stringency hybridization” refers generally to conditions that are about 10 °C or less from the thermal melting temperature Tmas determined under the solution condition for a defined polynucleotide sequence. In some embodiments, a high stringency condition refers to conditions that permit hybridization of only those nucleic acid sequences that form stable hybrids in 0.018M NaCI at 65 °C ( / .e., if a hybrid is not stable in 0.018M NaCI at 65 °C, it will not be stable under high stringency conditions, as contemplated herein). High stringency conditions can be provided, for example, by hybridization in conditions equivalent to 50% formamide, 5x Denhart's solution, 5xSSPE, 0.2% SDS at 42 °C, followed by washing in 0.1 xSSPE, and 0.1 % SDS at 65 °C. Another high stringency condition is hybridizing in conditions equivalent to hybridizing in 5X SSC containing 0.1% (w:v) SDS at 65 °C and washing in 0.1 x SSC containing 0.1 % SDS at 65 °C. Other high stringency hybridization conditions, as well as moderately stringent conditions, are described in the references cited above.“Heterologous” polynucleotide refers to any polynucleotide that is introduced into a host cell by laboratory techniques, and includes polynucleotides that are removed from a host cell, subjected to laboratory manipulation, and then reintroduced into a host cell.“Codon optimized” refers to changes in the codons of the polynucleotide encoding a protein to those preferentially used in a particular organism such that the encoded protein is efficiently expressed in the organism of interest. Although the genetic code is degenerate in that most amino acids arerepresented by several codons, called “synonyms” or “synonymous” codons, it is well known that codon usage by particular organisms is nonrandom and biased towards particular codon triplets. This codon usage bias may be higher in reference to a given gene, genes of common function or ancestral origin, highly expressed proteins versus low copy number proteins, and the aggregate protein coding regions of an organism's genome. In some embodiments, the polynucleotides encoding the transposase enzymes may be codon optimized for optimal production from the host organism selected for expression.As used herein, “preferred, optimal, high codon usage bias codons” refers interchangeably to codons that are used at higher frequency in the protein coding regions than other codons that code for the same amino acid. The preferred codons may be determined in relation to codon usage in a single gene, a set of genes of common function or origin, highly expressed genes, the codon frequency in the aggregate protein coding regions of the whole organism, codon frequency in the aggregate protein coding regions of related organisms, or combinations thereof. Codons whose frequency increases with the level of gene expression are typically optimal codons for expression. A variety of methods are known for determining the codon frequency (e.g., codon usage, relative synonymous codon usage) and codon preference in specific organisms, including multivariate analysis, for example, using cluster analysis or correspondence analysis, and the effective number of codons used in a gene (See e.g., GCG CodonPreference, Genetics Computer Group Wisconsin Package; CodonW, Peden, University of Nottingham; McInerney, Bioinform., 14:372-73

[1998] ; Stenico et al., Nucl. Acids Res., 222437-46

[1994] ; Wright, Gene 87:23-29

[1990] ). Codon usage tables are available for many different organisms (See e.g., Wada et al., Nucl. Acids Res., 20:2111 -2118

[1992] ; Nakamura et al., Nucl. Acids Res., 28:292

[2000] ; Duret, et al., supra; Henaut and Danchin, in Escherichia co / / and Salmonella, Neidhardt, et al. (eds.), ASM Press, Washington D.C., p. 2047-2066

[1996] ). The data source for obtaining codon usage may rely on any available nucleotide sequence capable of coding for a protein. These data sets include nucleic acid sequences actually known to encode expressed proteins (e.g., complete protein coding sequences-CDS), expressed sequence tags (ESTS), or predicted coding regions of genomic sequences (See e.g., Mount, Bioinformatics: Sequence and Genome Analysis, Chapter 8, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.

[2001] ; Uberbacher, Meth. Enzymol., 266:259-281

[1996] ; and Tiwari et al., Comput. Appl. Biosci., 13:263- 270

[1997] ).“Control sequence” is defined herein to include all components, which are necessary or advantageous for the expression of a polynucleotide and / or polypeptide of the present invention. Each control sequence may be native or foreign to the nucleic acid sequence encoding the polypeptide. Such control sequences include, but are not limited to, a leader, polyadenylation sequence, propeptide sequence, promoter, signal peptide sequence, and transcription terminator. At a minimum, the control sequences include a promoter, and transcriptional and translational stop signals. The control sequences may be provided with linkers for the purpose of introducing specific restriction sites facilitating ligation of the control sequences with the coding region of the nucleic acid sequence encoding a polypeptide.“Operably linked” is defined herein as a configuration in which a control sequence is appropriately placed ( / .e., in a functional relationship) at a position relative to a polynucleotide of interest such that the control sequence directs or regulates the expression of the polynucleotide and / or polypeptide of interest.“Promoter sequence” refers to a nucleic acid sequence that is recognized by a host cell for expression of a polynucleotide of interest, such as a coding sequence. The promoter sequence contains transcriptional control sequences, which mediate the expression of a polynucleotide of interest. The promoter may be any nucleic acid sequence which shows transcriptional activity in the host cell of choice including mutant, truncated, and hybrid promoters, and may be obtained from genes encoding extracellular or intracellular polypeptides either homologous or heterologous to the host cell.“Suitable reaction conditions” refer to those conditions in the biocatalytic reaction solution {e.g., ranges of enzyme loading, substrate loading, cofactor loading, temperature, pH, buffers, co-solvents, etc.) under which a transposase polypeptide of the present invention is capable of converting one or more substrate compounds to a product compound {e.g., ligation or annealing of an adapter or donor polynucleotide and a target polynucleotide to form an annealed or ligated polynucleotide consisting of the adapter and the optionally fragmented target polynucleotide). Exemplary “suitable reaction conditions” are provided in the present invention and illustrated by the Examples.“Composition” refers to a mixture or combination of one or more substances, wherein each substance or component of the composition retains its individual properties. As used herein, a biocatalytic composition refers to a combination of one or more substances useful for biocatalysis.“Loading”, such as in “compound loading” or “enzyme loading” or “cofactor loading” refers to the concentration or amount of a component in a reaction mixture at the start of the reaction. “Adapter loading” refers to binding of the transposase to the adapter polynucleotide.“Substrate” in the context of a biocatalyst mediated process refers to the compound or molecule acted on by the biocatalyst. For example, a transposase biocatalyst used in the synthesis processes disclosed herein acts on an adapter or donor polynucleotide and a target polynucleotide.“Product” in the context of a biocatalyst mediated process refers to the compound or molecule resulting from the action of the biocatalyst. For example, an exemplary product for a transposase biocatalyst used in a process disclosed herein is an annealed or ligated polynucleotide consisting of the adapter and the optionally fragmented target polynucleotide, as depicted in Figures 1 and 2.“Alkyl” refers to saturated hydrocarbon groups of from 1 to 18 carbon atoms inclusively, either straight chained or branched, more preferably from 1 to 8 carbon atoms inclusively, and most preferably 1 to 6 carbon atoms inclusively. An alkyl with a specified number of carbon atoms is denoted in parenthesis (e.g., (Ci-Ce)alkyl refers to an alkyl of 1 to 6 carbon atoms).“Alkenyl” refers to hydrocarbon groups of from 2 to 12 carbon atoms inclusively, either straight or branched containing at least one double bond but optionally containing more than one double bond.“Alkynyl” refers to hydrocarbon groups of from 2 to 12 carbon atoms inclusively, either straight or branched containing at least one triple bond but optionally containing more than one triple bond, and additionally optionally containing one or more double bonded moieties.“Heteroalkyl, “heteroalkenyl,” and heteroalkynyl,” refer respectively, to alkyl, alkenyl and alkynyl as defined herein in which one or more of the carbon atoms are each independently replaced with the same or different heteroatoms or heteroatomic groups. Heteroatoms and / or heteroatomic groups which can replace the carbon atoms include, but are not limited to -O-, -S-, -S-O-, -NR9-, -PH-, -S(O)-, -S(O)2-, -S(0) NR9-, -S(O)2NR9, and the like, including combinations thereof, where each R9 is independently selected from hydrogen, alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl.“Amino” refers to the group -NH2. Substituted amino refers to the group -NHRh, NRhRh, and NRhRhRh, where each Rhis independently selected from substituted or unsubstituted alkyl, cycloalkyl, cycloheteroalkyl, alkoxy, aryl, heteroaryl, heteroarylalkyl, acyl, alkoxycarbonyl, sulfanyl, sulfinyl, sulfonyl, and the like. Typical amino groups include, but are limited to, dimethylamino, diethylamino, trimethylammonium, triethylammonium, methylysulfonylamino, furanyl-oxy-sulfamino, and the like.“Aminoalkyl” refers to an alkyl group in which one or more of the hydrogen atoms are replaced with one or more amino groups, including substituted amino groups.“Aminocarbonyl” refers to -C(O)NH2. Substituted aminocarbonyl refers to -C(O)NRhRh, where the amino group NRhRhis as defined herein.“Oxy” refers to a divalent group -O-, which may have various substituents to form different oxy groups, including ethers and esters.“Alkoxy” or “alkyloxy” are used interchangeably herein to refer to the group -ORZ, wherein Rzis an alkyl group, including optionally substituted alkyl groups.“Carboxy” refers to -COOH.“Carbonyl” refers to -C(O)-, which may have a variety of substituents to form different carbonyl groups including acids, acid halides, aldehydes, amides, esters, and ketones.“Carboxyalkyl” refers to an alkyl in which one or more of the hydrogen atoms are replaced with one or more carboxy groups.“Aminocarbonylalkyl” refers to an alkyl substituted with an aminocarbonyl group, as defined herein.“Halogen” or “halo” refers to fluoro, chloro, bromo and iodo.“Haloalkyl” refers to an alkyl group in which one or more of the hydrogen atoms are replaced with a halogen. Thus, the term “haloalkyl” is meant to include monohaloalkyls, dihaloalkyls, trihaloalkyls, etc. up to perhaloalkyls. For example, the expression “(Ci - C2) haloalkyl” includes 1 -fluoromethyl, difluoromethyl, trifluoromethyl, 1 -fluoroethyl, 1 ,1 -difluoroethyl, 1 ,2-difluoroethyl, 1 ,1 ,1 trifluoroethyl, perfluoroethyl, etc.“Hydroxy” refers to -OH.“Hydroxyalkyl” refers to an alkyl group in which in which one or more of the hydrogen atoms are replaced with one or more hydroxy groups.“Thiol” or “sulfanyl” refers to -SH. Substituted thiol or sulfanyl refers to -S-Rh, where Rhis an alkyl, aryl or other suitable substituent.“Sulfonyl” refers to -SO2-. Substituted sulfonyl refers to -SO2-Rh, where Rhis an alkyl, aryl or other suitable substituent.“Alkylsulfonyl" refers to -SO2-RZ, where Rzis an alkyl, which can be optionally substituted. Typical alkylsulfonyl groups include, but are not limited to, methylsulfonyl, ethylsulfonyl, n-propylsulfonyl, and the like.“Phosphate” as used herein refers to a functional group comprised of an orthophosphate ion (phosphorous atom covalently linked to four oxygen atoms). The orthophosphate ion is commonly found with one or more hydrogen atoms or organic groups.“Phosphorylated” as used herein refers to the addition or presence of one of more phosphoryl groups (phosphorous atom covalently linked to the three oxygen atoms).“Optionally substituted” as used herein with respect to the foregoing chemical groups means that positions of the chemical group occupied by hydrogen can be substituted with another atom (unless otherwise specified) exemplified by, but not limited to carbon, oxygen, nitrogen, or sulfur, or a chemical group, exemplified by, but not limited to, hydroxy, oxo, nitro, methoxy, ethoxy, alkoxy, substituted alkoxy, trifluoromethoxy, haloalkoxy, fluoro, chloro, bromo, iodo, halo, methyl, ethyl, propyl, butyl, alkyl, alkenyl, alkynyl, substituted alkyl, trifluoromethyl, haloalkyl, hydroxyalkyl, alkoxyalkyl, thio, alkylthio, acyl, carboxy, alkoxycarbonyl, carboxamido, substituted carboxamido, alkylsulfonyl, alkylsulfinyl, alkylsulfonylamino, sulfonamido, substituted sulfonamido, cyano, amino, substituted amino, alkylamino, dialkylamino, aminoalkyl, acylamino, amidino, amidoximo, hydroxamoyl, phenyl, aryl, substituted aryl, aryloxy, arylalkyl, arylalkenyl, arylalkynyl, pyridyl, imidazolyl, heteroaryl, substituted heteroaryl, heteroaryloxy, heteroarylalkyl, heteroarylalkenyl, heteroarylalkynyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloalkyl, cycloalkenyl, cycloalkylalkyl, substituted cycloalkyl, cycloalkyloxy, pyrrolidinyl, piperidinyl, morpholino, heterocycle, (heterocycle)oxy, and (heterocycle)alkyl; where preferred heteroatoms are oxygen, nitrogen, and sulfur. Additionally, where open valences exist on these substitute chemical groups they can be further substituted with alkyl, cycloalkyl, aryl, heteroaryl, and / or heterocycle groups, that where these open valences exist on carbon they can be further substituted by halogen and by oxygen-, nitrogen-, or sulfur-bonded substituents, and where multiple such open valences exist, these groups can be joined to form a ring, either by direct formation of a bond or by formation of bonds to a new heteroatom, preferably oxygen, nitrogen, or sulfur. It is further contemplated that the above substitutions can be made provided that replacing the hydrogen with the substituent does not introduce unacceptable instability to the molecules of the present invention, and is otherwise chemically reasonable. One of ordinary skill in the art would understand that with respect to any chemical group described as optionally substituted, only sterically practical and / or synthetically feasible chemical groups are meant to be included. “Optionally substituted” as used herein refers to all subsequent modifiers in a term or series of chemical groups. For example, in the term "optionally substituted arylalkyl,” the “alkyl” portion and the “aryl” portion of the molecule may or may not be substituted, and for the series “optionally substituted alkyl, cycloalkyl, aryl and heteroaryl,” the alkyl, cycloalkyl, aryl, and heteroaryl groups, independently of the others, may or may not be substituted.“Reaction” as used herein refers to a process in which one or more substances or compounds or substrates is converted into one or more different substances, compounds, or processes.Preparation of Next Generation Sequencing Libraries Using Engineered TransposasesTransposases with improved activity in cleaving or fragmenting polynucleotides and ligating or tagging polynucleotides are necessary to overcome the limitations of existing NGS library preparation methods. Improved transposases with increased polynucleotide fragmentation or cleavage activity,increased thermostability, increased activity at elevated temperatures, increased soluble expression, decreased inhibition, increased adapter loading or adapter ligation (i.e. tagging), increased binding to a target polynucleotide, and / or decreased sequence insertion bias compared to another wild-type or engineered transposase find utility in library preparation for NGS, but are not limited to these uses. The improved transposases of the present disclosure find use in any application involving cleaving or fragmenting double-stranded polynucleotides and / or ligating adapters or tagging polynucleotides, including applications where a donor polynucleotide is transferred into a target polynucleotide.The present invention provides novel transposases that have improved activity in the fragmentation or cleavage of double stranded DNA and the ligation of adapters or tagging of the DNA fragments. The transposases of the present disclosure have increased polynucleotide fragmentation or cleavage activity, increased thermostability, increased activity at elevated temperatures, increased soluble expression, decreased inhibition, increased adapter loading or adapter ligation (i.e. tagging), increased binding to a target polynucleotide, and / or decreased sequence insertion bias as compared to another wild-type transposase or engineered transposase. The engineered polypeptides of the present disclosure are variants of SEQ ID NO: 2, a chimeric protein based an IS4 transposase from Parashewanella curva or SEQ ID NO: 5704, a modified hyperactive Tn5 transposase, and designed to recognize the19 bp Tn5 mosaic end (ME) transposon donor recognition sequence. These engineered transposases are capable of improved preparation of NGS libraries from polynucleotide samples.In nature, transposases facilitate the transposition of fragments of DNA (transposons) from a donor strand to a host strand at sites flanked by short inverted-repeat sequences or end sequences (ESs). The transposition mechanism is initiated by the binding of two transposases to donor DNA ESs to form the dimeric synaptic complex, which catalyzes cleavage of the donor DNA. The transposome (dimeric transposase / DNA complex) then binds to the target DNA to transfer the transposon DNA to the target DNA.Transposase-mediated NGS library preparation, depicted in Figure 1 , is similar to naturally- occurring transposition, except that the donor DNA or polynucleotide consists of two, separate adapter sequences rather than one continuous loop of donor DNA or polynucleotide. These adapter sequences are bound to transposases in a loading step that replaces the initial donor DNA or polynucleotide binding and cleavage step. Following adapter loading, the dimerized transposome proceeds to bind the target DNA or polynucleotide sample. Cleavage of the target polynucleotide and ligation of the adapter sequences results in fragmentation of the target DNA or polynucleotide and ligation of sequencing adapters to the target fragments.The resulting DNA or polynucleotide fragments with ligated adapters comprise the next generation sequencing library. An ideal library preparation method would generate random fragments in the polynucleotide sequence, without bias toward the sequence composition of the target site. Compared to traditional library preparation methods, transposase-mediated fragmentation has lower sequence insertion bias. However, certain transposases are known to have sequence insertion bias. As an example, Tn5 has a slight G / C insertion bias.In some embodiments, the engineered transposases of the present disclosure have reduced sequence insertion bias or increased sequence promiscuity as compared to a wild-type or reference transposase.In addition to containing the ME or ES sequences for target binding, adapters may comprise sequencing primers or other sequences used to identify or select fragments or groups of fragments from pooled samples. The adapters may encode primer binding sites for next generation sequencing library amplification, binding sites for sequencing primers, sample indexes, or other functional sequences related to a specific application. In some embodiments, the engineered transposases of the present disclosure have increased adapter loading or binding, as compared to a wild-type or reference transposase.In some embodiments, further processing steps are used for library preparation, including gap filling, normalization, pooling of tagged fragment samples, and / or ligation of additional adapters to create dual-tagged libraries. A variety of parameters may be manipulated to create sequencing libraries with various characteristics. For example, varying the concentration of transposase and adapters can change the fragment length. Copy number and sample coverage may also be influenced or controlled using methods known to those of skill in the art.As described further herein, the engineered transposase polypeptides of the current disclosure exhibit one of more improved properties in the transposase-mediated DNA fragmentation and adapter ligation process depicted in Figure 1.In some embodiments, the present invention provides an engineered transposase polypeptide comprising an amino acid sequence having at least 60% sequence identity to an amino acid reference sequence of SEQ ID NO: 2 or SEQ ID NO: 5704 and further comprising one or more amino acid residue differences as compared to the reference amino acid sequence, wherein the engineered transposase polypeptide has increased polynucleotide fragmentation or cleavage activity, increased thermostability, increased activity at elevated temperatures, increased soluble expression, decreased inhibition, increased adapter loading or adapter ligation (i.e. tagging), increased binding to a target polynucleotide, and / or decreased sequence insertion bias as compared to another wild-type transposase or engineered transposase.In particular, the engineered transposase polypeptides of the present disclosure have been engineered for efficient fragmentation or cleavage of DNA and the ligation of adapters in the process described above.A variety of suitable reaction conditions are known to those skilled in the art, as detailed below and in the Examples.Engineered Transposase PolypeptidesThe present invention provides engineered transposase polypeptides useful in preparation of next generation sequencing libraries, as well as compositions and methods of utilizing these engineered polypeptides.The present invention provides transposase polypeptides, polynucleotides encoding the polypeptides, methods of preparing the polypeptides, and methods for using the polypeptides. Where thedescription relates to polypeptides, it is to be understood that it can describe the polynucleotides encoding the polypeptides.Suitable reaction conditions under which the above-described improved properties of the engineered polypeptides carry out the desired reaction can be determined with respect to concentrations or amounts of polypeptide, substrate, co-substrate, buffer, solvent, pH, conditions including temperature and reaction time, and / or conditions with the polypeptide immobilized on a solid support, as further described below and in the Examples.In some embodiments, exemplary engineered transposases comprise an amino acid sequence that has one or more residue differences as compared to SEQ ID NO: 2 at the residue positions indicated in Tables 5.2, 6.2, 7.2, 8.2, 9.2, 10.2, 10.3, 10.5, 11.2, 11.4, 11.5, 12.2, 12.3, 12.5, 12.6, 13.2, 13.3, 13.4,14.2, 14. 3, 14.5, 14.6, 14.8, 14.9, 15.1 , 16.2, 16.3, 17.2, 17.3, 18.2, 19.2, 19.3, 20.2, 20.3, 21.2, 21.3,22.2, 23.2, 24.2, 25.2, 26.2, 27.2, 28.2, 29.2, 30.2, 31 .2, 32.2, 33.2, 34.2, 35.1 . In some embodiments, exemplary engineered transposases comprise an amino acid sequence that has one or more residue differences as compared to SEQ ID NO: 5704 at the residue positions indicated in Tables 36.1 and 36.3.The structure and function information for the exemplary engineered polypeptides of the present invention are based on the fragmentation or cleavage of double stranded DNA and the ligation or tagging of the DNA fragments, the results of which are shown below in Tables 5.2, 6.2, 7.2, 8.2, 9.2, 10.2, 10.3, 10.5, 11.2, 11.4, 11.5, 12.2, 12.3, 12.5, 12.6, 13.2, 13.3, 13.4, 14.2, 14. 3, 14.5, 14.6, 14.8, 14.9, 15.1 ,16.2, 16.3, 17.2, 17.3, 18.2, 19.2, 19.3, 20.2, 20.3, 21 .2, 21 .3, 22.2, 23.2, 24.2, 25.2, 26.2, 27.2, 28.2,29.2, 30.2, 31 .2, 32.2, 33.2, 34.2, 35.1 , 36.1 , and 36.3, as further described in the Examples. The odd numbered sequence identifiers (i.e., SEQ ID NOs) in these Tables refer to the nucleotide sequence encoding the amino acid sequence provided by the even numbered SEQ ID NOs in these Tables. Exemplary sequences are provided in the electronic sequence listing file accompanying this invention, which is hereby incorporated by reference herein. The amino acid residue differences are based on comparison to the reference sequence of SEQ ID NOs: 2, 4, 156, 302, 338, 606, 972, 1082, 1218, 1580, 1598, 1600, 1604, 1720, 2368, 2426, 2540, 3402, 3662, 3788, 4150, 4278, 4642, 4984, 5196, 5286, 5412, 5422, and / or 5704, as indicated.Proteins based on an IS4 transposase from Parashewanella curva or a Tn5 transposase with the ability to recognize the 19 bp Tn5 mosaic end (ME) transposon donor recognition sequence (SEQ ID NO: 2 or SEQ ID NO: 5704, respectively) were designed in silico, verified to have transposase activity, and selected for evolution. The transposase polypeptides of the present disclosure are engineered variants of SEQ ID NO: 2 or SEQ ID NO: 5704, which also comprise a C-terminal 6-histidine tag following a GGSG peptide spacer (SEQ ID NO: 5777).The polypeptides of the present disclosure have residue differences that result in improved properties, including increased polynucleotide fragmentation or cleavage activity, increased thermostability, increased activity at elevated temperatures, increased soluble expression, decreased inhibition, increased adapter loading or adapter ligation (i.e. tagging), increased binding to a target polynucleotide, and / or decreased sequence insertion bias as compared to another wild-type transposase or engineered transposase.The activity of each engineered transposase relative to the reference polypeptide of SEQ ID NOs: 2, 4, 156, 302, 338, 606, 972, 1082, 1218, 1580, 1598, 1600, 1604, 1720, 2368, 2426, 2540, 3402, 3662, 3788, 4150, 4278, 4642, 4984, 5196, 5286, 5412, 5422, and / or 5704 was determined as conversion of the substrates described in the Examples herein. In some embodiments, a shake flask purified enzyme (SFP) is used to assess the properties of the engineered transposases, the results of which are provided in the Examples.In some embodiments, the specific enzyme properties are associated with the residues differences as compared to SEQ ID NOs: 2, 4, 156, 302, 338, 606, 972, 1082, 1218, 1580, 1598, 1600, 1604, 1720, 2368, 2426, 2540, 3402, 3662, 3788, 4150, 4278, 4642, 4984, 5196, 5286, 5412, 5422, and / or 5704 at the residue positions indicated herein. In some embodiments, residue differences affecting polypeptide expression can be used to increase expression of the engineered transposases.In light of the guidance provided herein, it is further contemplated that any of the exemplary engineered polypeptides comprising the polypeptide sequences of SEQ ID NOs: 2, 4, 156, 302, 338, 606, 972, 1082, 1218, 1580, 1598, 1600, 1604, 1720, 2368, 2426, 2540, 3402, 3662, 3788, 4150, 4278, 4642, 4984, 5196, 5286, 5412, 5422, and / or 5704 find use as the starting amino acid sequence for synthesizing other transposase polypeptides, for example by subsequent rounds of evolution that incorporate new combinations of various amino acid differences from other polypeptides in Tables 5.2, 6.2, 7.2, 8.2, 9.2,10.2, 10.3, 10.5, 11.2, 11.4, 11.5, 12.2, 12.3, 12.5, 12.6, 13.2, 13.3, 13.4, 14.2, 14. 3, 14.5, 14.6, 14.8,14.9, 15.1 , 16.2, 16.3, 17.2, 17.3, 18.2, 19.2, 19.3, 20.2, 20.3, 21.2, 21.3, 22.2, 23.2, 24.2, 25.2, 26.2,27.2, 28.2, 29.2, 30.2, 31 .2, 32.2, 33.2, 34.2, 35.1 , 36.1 , and 36.3, and other residue positions described herein. Further improvements may be generated by including amino acid differences at residue positions that had been maintained as unchanged throughout earlier rounds of evolution.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 2 and one or more residue differences as compared to SEQ ID NO: 2, selected from 10, 13, 24, 35, 39, 44, 46, 51 , 54, 55, 62 / 361 , 72, 75, 82, 89, 92, 107, 128, 137, 164, 167, 177 / 359, 191 , 192, 193, 194, 201 , 217, 281 , 289, 291 , 291 / 340, 305, 309, 310, 317, 331 , 337, 340, 346, 352, 353, 357, 358, 359, 362, 371 , 396, 414 / 435, 415, 428, 437, 443, 447, and 448. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 2 and one or more residue differences as compared to SEQ ID NO: 2, selected from 10A, 13E, 24L, 35L, 39P, 44P, 46A, 51 E, 54T, 55K, 62D / 361 H, 72L, 75H, 82R, 89V, 92L, 107G, 128A, 137L, 164E, 167R, 177I / 359A, 191 Y, 192E, 193L, 194F, 201 G, 217G, 2811, 289P, 291 G, 291 Y / 340L, 305V, 309E, 310D, 317W, 331 L, 337I, 340L, 346D, 352L, 353A, 357I, 358M, 358V, 359A, 362I, 371 L, 396I, 414D / 435L, 415P, 428I, 437R, 443P, 447V, and 448L. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 2 and one or more residue differences as compared to SEQ ID NO: 2, selected from S10A, H13E, R24L, M35L, S39P, T44P, S46A, K51 E, Q54T, E55K,R62D / R361 H, R72L, G75H, L82R, I89V, I92L, D107G, L128A, I137L, D164E, H167R, M177I / G359A, Q191Y, S192E, Y193L, L194F, N201 G, S217G, V281 I, K289P, D291 G, D291Y / Q340L, I305V, S309E, E310D, 1317W, W331 L, A337I, Q340L, R346D, V352L, S353A, F357I, I358M, I358V, G359A, L362I, P371 L, F396I, N414D / M435L, G415P, L428I, S437R, A443P, A447V, and I448L.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 4 and one or more residue differences as compared to SEQ ID NO: 4, selected from 4, 5, 6, 8, 10, 10 / 317, 10 / 317 / 358, 10 / 317 / 359, 10 / 346, 10 / 346 / 359, 10 / 358, 10 / 359, 12, 12 / 373, 13, 15, 16, 18, 19, 26, 30, 30 / 154, 31 , 34, 35, 37, 38, 68, 70, 73, 74, 93, 106, 118, 126, 134, 135, 143, 146, 154 / 453, 163, 182, 207 / 286, 208, 226, 237, 246, 247, 259, 262, 263, 264, 280, 282, 286, 287, 288, 289, 291 , 292, 293, 298, 305, 309, 310, 313, 317, 317 / 346, 317 / 359, 317 / 445, 320, 346, 351 , 355, 356, 358, 359, 360, 368, 369, 410, 411 , 422, 426, 427, 428, 446, 448, 453, and 454. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 4 and one or more residue differences as compared to SEQ ID NO: 4, selected from 4H, 5R, 6A, 8D, 10A, 10A / 317W, 10A / 317W / 358V, 10A / 317W / 359A, 10A / 346D, 10A / 346D / 359A, 10A / 358V, 10A / 359A, 12S, 12S / 373C, 13V, 15S, 16S, 18A, 19L, 26A, 30N, 30N / 154V, 31 V, 34Q, 35L, 37K, 38Y, 68A, 70A, 73K, 74A, 93E, 106E, 118S, 126V, 134F, 135R, 143E, 146M, 154P / 453E, 163L, 182A, 207C / 286I, 208S, 226K, 237S, 246K, 247R, 259S, 262L, 263R, 264S, 280A, 282L, 286I, 287N, 288P, 289P, 291 G, 292E, 293T, 298L, 305V, 309A, 310Q, 313R, 317W, 317W / 346D, 317W / 359A, 317W / 445L, 320H, 346D, 351 M, 355L, 356S, 358V, 359A, 360V, 368S, 369F, 410G, 411 K, 422Q, 426M, 427A, 428I, 446G, 448L, 453E, and 454A. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 4 and one or more residue differences as compared to SEQ ID NO: 4, selected from F4H, N5R, T6A, Q8D, S10A, S10A / I317W, S10A / I317W / I358V, S10A / I317W / G359A, S10A / R346D, S10A / R346D / G359A, S10A / I358V, S10A / G359A, E12S, E12S / Y373C, H13V, E15S, K16S, N18A, T19L, K26A, E30N, E30N / A154V, T31 V, D34Q, M35L, R37K, C38Y, P68A, V70A, C73K, S74A, D93E, S106E, A118S, T126V, T134F, K135R, D143E, C146M, A154P / D453E, A163L, S182A, R207C / T286I, A208S, E226K, D237S, S246K, K247R, K259S, V262L, K263R, V264S, V280A, Y282L, T286I, T287N, S288P, K289P, D291 G, M292E, E293T, V298L, I305V, S309A, E310Q, H313R, 1317W, I317W / R346D, I317W / G359A, I317W / W445L, A320H, R346D, A351 M, C355L, A356S, I358V, G359A, I360V, A368S, I369F, P410G, R411 K, K422Q, Q426M, S427A, L428I, E446G, I448L, D453E, and T454A.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 156 and one or more residue differences as compared to SEQ ID NO: 156, selected from 18 / 73 / 74 / 259, 18 / 73 / 286 / 289 / 411 , 18 / 74 / 118 / 259 / 286 / 289 / 411 , 18 / 74 / 259, 18 / 74 / 259 / 286 / 289, 18 / 74 / 259 / 286 / 411 , 18 / 118 / 259, 18 / 259 / 286 / 289 / 411 , 18 / 259 / 289 / 411 , 18 / 289 / 411 ,73 / 74 / 259 / 286 / 289, 73 / 74 / 259 / 289, 73 / 259 / 286, 74 / 259, 74 / 259 / 289, 74 / 286 / 289 / 411 , 259, 259 / 286 / 289, and 289. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95% 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 156 and one or more residue differences as compared to SEQ ID NO: 156, selected from 18A / 73K / 74A / 259S, 18A / 73K / 286I / 289P / 411 K, 18A / 74A / 118S / 259S / 286I / 289P / 411 K, 18A / 74A / 259S,18A / 74A / 259S / 286I / 289P, 18A / 74A / 259S / 286I / 411 K, 18A / 118S / 259S, 18A / 259S / 286I / 289P / 411 K, 18A / 259S / 289P / 411 K, 18A / 289P / 411 K, 73K / 74A / 259S / 286I / 289P, 73K / 74A / 259S / 289P, 73K / 259S / 286I, 74A / 259S, 74A / 259S / 289P, 74A / 286I / 289P / 411 K, 259S, 259S / 286I / 289P, and 289P. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 156 and one or more residue differences as compared to SEQ ID NO: 156, selected from N18A / C73K / S74A / K259S,N18A / C73K / T286I / K289P / R411 K, N18A / S74A / A118S / K259S / T286I / K289P / R411 K, N18A / S74A / K259S, N18A / S74A / K259S / T286I / K289P, N18A / S74A / K259S / T286I / R411 K, N18A / A118S / K259S, N18A / K259S / T286I / K289P / R411 K, N18A / K259S / K289P / R411 K, N18A / K289P / R411 K, C73K / S74A / K259S / T286I / K289P, C73K / S74A / K259S / K289P, C73K / K259S / T286I, S74A / K259S, S74A / K259S / K289P, S74A / T286I / K289P / R411 K, K259S, K259S / T286I / K289P, and K289P.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 302 and one or more residue differences as compared to SEQ ID NO: 302, selected from 16 / 19 / 355 / 427, 16 / 37 / 146 / 286 / 287 / 289 / 310 / 313 / 355, 16 / 146 / 226 / 287 / 289 / 355, 19 / 37 / 146 / 286 / 287 / 310 / 355, 19 / 135 / 286 / 287 / 289 / 355, 19 / 182 / 355, 37 / 70 / 226 / 310 / 313 / 355, 37 / 74 / 310 / 355 / 427, 37 / 135 / 146 / 182 / 226 / 286 / 287 / 355 / 368, 37 / 146 / 226 / 289 / 427, 37 / 146 / 287 / 289 / 427, 38, 41 , 42, 43, 44, 45, 47, 48, 50, 53, 54, 55, 70 / 74 / 226 / 286 / 287 / 289 / 368 / 427, 70 / 74 / 355 / 368, 70 / 135 / 146 / 150 / 182 / 286 / 289 / 313 / 427, 74 / 310 / 313 / 355, 74 / 310 / 355, 74 / 355 / 427, 101 , 103, 117, 135 / 226 / 286 / 289 / 355 / 368, 135 / 355, 146, 146 / 150 / 287 / 289 / 310 / 355, 146 / 182 / 355, 146 / 226 / 286 / 355, 146 / 286 / 287 / 289 / 355 / 427, 146 / 289, 146 / 427, 150 / 226 / 310 / 313, 182 / 226 / 355, 182 / 355, 244, 247, 249, 251 , 287 / 355 / 427, 300, 317 / 411 , 355, 393, 397, 398, 399, 401 , 405, 407, 408, 409, 411 , 413, 426, 429, 431 , 439, 444, 446, 450, 453, 454, 457, 459, 460, 464, 466, 467, 469, 470, 471 , 472, 474, 475, 476, and 478. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 302 and one or more residue differences as compared to SEQ ID NO: 302, selected from 16S / 19L / 355L / 427A, 16S / 37K / 146M / 286I / 287N / 289P / 31 OQ / 313R / 355L, 16S / 146M / 226K / 287N / 289P / 355L, 19L / 37K / 146M / 286I / 287N / 310Q / 355L, 19L / 135R / 286I / 287N / 289P / 355L, 19L / 182A / 355L, 37K / 70A / 226K / 31 OQ / 313R / 355L, 37K / 74S / 310Q / 355L / 427A, 37K / 135R / 146M / 182A / 226K / 286I / 287N / 355L / 368S, 37K / 146M / 226K / 289P / 427A, 37K / 146M / 287N / 289P / 427A, 38F, 38N, 41 R, 42G, 43L, 43V, 44G, 44S, 44V, 45M, 45V, 47M, 47S, 48D,50D, 53D, 53L, 54H, 54S, 54V, 55M, 70A / 74S / 226K / 286I / 287N / 289P / 368S / 427A, 70A / 74S / 355L / 368S, 70A / 135R / 146M / 150H / 182A / 286I / 289P / 313R / 427A, 74S / 31 OQ / 313R / 355L, 74S / 310Q / 355L, 74S / 355L / 427A, 101 S, 103M, 103R, 117R, 135R / 226K / 286I / 289P / 355L / 368S, 135R / 355L, 146M, 146M / 150H / 287N / 289P / 310Q / 355L, 146M / 182A / 355L, 146M / 226K / 286I / 355L, 146M / 286I / 287N / 289P / 355L / 427A, 146M / 289P, 146M / 427A, 150H / 226K / 31 OQ / 313R, 182A / 226K / 355L, 182A / 355L, 244I, 244T, 247L, 249F, 249I, 249L, 249S, 249T, 249V, 249Y, 251 L, 287N / 355L / 427A, 300F, 317R / 411 N, 355L, 393A, 397S, 398V, 399V, 401 V, 405F, 405V, 407L, 408R, 409G, 411 D, 413Y, 426A, 426H, 426M, 426S, 426V, 426W, 429G, 431 M, 439G, 444G, 446K, 446Q, 450L, 450M, 450T, 453G, 453H, 453R, 453T, 454L, 454R, 457A, 459M, 460T, 464L, 464M, 466R, 467G, 469F, 470M, 470N, 470R, 470V, 470Y, 471 A, 471 S, 472R, 474R, 474S, 475P, 476D, 476L, 476S, 476T, 478C, 478G, 478L, and 478V. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 302 and one or more residue differences as compared to SEQ ID NO: 302, selected from K16S / T19L / C355L / S427A, K16S / R37K / C146M / T286I / T287N / K289P / E310Q / H313R / C355L,K16S / C146M / E226K / T287N / K289P / C355L, T19L / R37K / C146M / T286I / T287N / E310Q / C355L, T19L / K135R / T286I / T287N / K289P / C355L, T19L / S182A / C355L, R37K / V70A / E226K / E310Q / H313R / C355L, R37K / A74S / E310Q / C355L / S427A, R37K / K135R / C146M / S182A / E226K / T286I / T287N / C355L / A368S, R37K / C146M / E226K / K289P / S427A, R37K / C146M / T287N / K289P / S427A, C38F, C38N, K41 R, S42G, I43L, I43V, T44G, T44S, T44V, I45M, I45V, C47M, C47S, Q48D, S50D, M53D, M53L, Q54H, Q54S, Q54V, E55M, V70A / A74S / E226K / T286I / T287N / K289P / A368S / S427A, V70A / A74S / C355L / A368S, V70A / K135R / C146M / N150H / S182A / T286I / K289P / H313R / S427A, A74S / E310Q / H313R / C355L, A74S / E310Q / C355L, A74S / C355L / S427A, K101 S, Q103M, Q103R, K117R, K135R / E226K / T286I / K289P / C355L / A368S, K135R / C355L, C146M,C146M / N150H / T287N / K289P / E310Q / C355L, C146M / S182A / C355L, C146M / E226K / T286I / C355L, C146M / T286I / T287N / K289P / C355L / S427A, C146M / K289P, C146M / S427A, N150H / E226K / E310Q / H313R, S182A / E226K / C355L, S182A / C355L, V244I, V244T, K247L, K249F, K249I, K249L, K249S, K249T, K249V, K249Y, K251 L, T287N / C355L / S427A, L300F, W317R / R411 N, C355L, E393A, D397S, E398V, D399V, W401 V, M405F, M405V, M407L, K408R, K409G, R411 D, W413Y, Q426A, Q426H, Q426M, Q426S, Q426V, Q426W, A429G, L431 M, R439G, S444G, E446K, E446Q, E450L, E450M, E450T, D453G, D453H, D453R, D453T, T454L, T454R, S457A, V459M, V460T, V464L, V464M, K466R, E467G, L469F, A470M, A470N, A470R, A470V, A470Y, D471 A, D471 S, G472R, L474R, L474S, L475P, G476D, G476L, G476S, G476T, S478C, S478G, S478L, and S478V.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 338 and one or more residue differences as compared to SEQ ID NO: 338, selected from 16, 16 / 313, 16 / 409 / 411 / 427, 37 / 150 / 408, 37 / 313 / 317 / 408 / 411 , 37 / 317 / 405 / 408, 37 / 317 / 405 / 426, 37 / 405 / 409 / 427, 150, 150 / 313 / 405 / 411 , 150 / 405 / 406, 150 / 408, 150 / 408 / 411 , 226 / 405, 317 / 460, and 408 / 426. In some embodiments, the engineered transposase polypeptide comprises anamino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 338 and one or more residue differences as compared to SEQ ID NO: 338, selected from 16S, 16S / 313R, 16S / 409D / 411 N / 427M, 37K / 150N / 408G, 37K / 313R / 317R / 408G / 411 N, 37K / 317R / 405V / 408G,37K / 317R / 405V / 426M, 37K / 405V / 409D / 427M, 150N, 150N / 313R / 405V / 411 N, 150N / 405V / 406S, 150N / 408G, 150N / 408G / 411 N, 226K / 405V, 317R / 460L, and 408G / 426M. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 338 and one or more residue differences as compared to SEQ ID NO: 338, selected from K16S, K16S / H313R, K16S / K409D / R411 N / S427M, R37K / H150N / K408G, R37K / H313R / W317R / K408G / R411 N, R37K / W317R / M405V / K408G, R37K / W317R / M405V / Q426M, R37K / M405V / K409D / S427M, H150N, H150N / H313R / M405V / R411 N, H150N / M405V / K406S, H150N / K408G, H150N / K408G / R411 N, E226K / M405V, W317R / V460L, and K408G / Q426M.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 606 and one or more residue differences as compared to SEQ ID NO: 606, selected from 12, 13, 18, 19, 22, 26, 317262, 34, 60, 79, 80, 81 , 82, 92, 106, 107, 113, 115, 132, 134, 136,140, 143, 152, 160, 163, 167, 168, 175, 179, 183, 195, 199, 201 , 202, 215, 216, 218, 220, 223, 226, 233,235, 236, 237, 243, 255, 259, 261 , 262, 266, 268, 270, 273, 282, 290, 292, 293, 309, 319, 320, 326, 343,346, 365, 370, 374, 380, 382, 386, 389, 390, 408, 412, 414, 415, 424, and 442. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 606 and one or more residue differences as compared to SEQ ID NO: 606, selected from 12M, 131, 18Q, 19L, 22S, 22T, 26R, 31 M / 262A, 34G, 60L, 79Y, 80T, 81 T, 82Q, 92L, 92T, 106Q, 107G, 107H, 107R, 113K, 115R, 132G, 132R, 134E, 134G, 136L, 140C, 140L, 140V, 143G, 143R, 143V, 152S, 160Q, 160T, 163L, 163T, 167R, 168Q, 175T, 179G, 183L, 195G, 199R, 199T, 199W, 201 G, 201 Q, 202M, 215R, 215V, 216E, 218G, 220R, 223R, 223V, 226L, 226Q, 233R, 235V, 236T, 237G, 237S, 243L, 243M, 255R, 259A, 261 E, 262L, 266P, 268A, 270R, 273K, 273P, 273T, 282G, 290A, 290R, 290T, 292W, 293V, 309G, 319R, 320R, 320S, 326A, 326L, 343G, 343R, 346G, 346N, 365I, 370F, 374L, 380R, 382A, 382R, 386D, 389G, 390R, 408A, 408R, 412A, 412H, 4121, 412R, 414R, 415S, 424V, and 442K. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 606 and one or more residue differences as compared to SEQ ID NO: 606, selected from E12M, H13I, A18Q, T19L, P22S, P22T, K26R, T31 M / V262A, D34G, F60L, T79Y, A80T, V81T, L82Q, I92L, I92T, S106Q, D107G, D107H, D107R, S113K, K115R, F132G, F132R, T134E, T134G, T136L, I140C, I140L, 1140V, D143G, D143R, D143V, E152S, G160Q, G160T, A163L, A163T, H167R, F168Q, D175T, R179G, V183L, Q195G, S199R, S199T, S199W, N201 G, N201 Q, E202M, L215R, L215V, D216E, E218G, P220R, E223R, E223V, E226L, E226Q, A233R, T235V, I236T, D237G, D237S, V243L, V243M, A255R, S259A,S261 E, V262L, A266P, T268A, K270R, D273K, D273P, D273T, Y282G, K290A, K290R, K290T, M292W, E293V, S309G, T319R, A320R, A320S, E326A, E326L, E343G, E343R, R346G, R346N, L365I, T370F, Y374L, L380R, D382A, D382R, E386D, S389G, Q390R, G408A, G408R, G412A, G412H, G412I, G412R, N414R, G415S, A424V, and I442K.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 606 and one or more residue differences as compared to SEQ ID NO: 606, selected from 2, 12, 13, 15, 16, 18, 19, 22, 26, 30, 31 / 262, 34, 51 , 60, 69, 79, 82, 84, 87, 88, 106, 107, 113, 114, 115, 128, 132, 134, 135, 136, 140, 143, 146, 150, 152, 160, 163, 167, 168, 175, 179, 183, 195,198, 199, 201 , 215, 217, 218, 219, 220, 223, 224, 226, 227, 231 , 233, 235, 236, 243, 254, 255, 259, 261 ,266, 267, 268, 270, 272, 273, 275, 277, 279, 282, 287, 289, 290, 291 , 292, 293, 304, 307, 309, 319, 320,343, 344, 346, 355, 365, 370, 373, 380, 382, 386, 389, 390, 408, 412, 414, 415, 418, 423, 424, and 468.In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 606 and one or more residue differences as compared to SEQ ID NO: 606, selected from 2A, 12L, 12M, 131, 13N, 13Y, 15R, 16S, 18Q, 19L, 22S, 22T, 26R, 30H, 31 M / 262A, 34G, 51 R, 60L, 69A, 69L, 79Y, 82Q, 84R, 87T, 88L, 106Q, 107G, 107H, 107R, 113E, 114P, 115S, 128F, 132G, 132R, 134E, 134G, 134L, 135T, 136L, 140C, 140L, 140V, 143G, 143R, 143V, 146K, 150R, 152S, 160Q, 160T, 163L, 163T, 167R, 168Q, 175T, 175V, 179G, 183L, 195G, 195S, 198S, 199R, 199T, 199W, 201 G, 201 Q, 215P, 215V, 217F, 217G, 218G, 219S, 220R, 223R, 223V, 224A, 224S, 226L, 227Y, 231 V, 233R, 235A, 235V, 236T, 243L, 243M, 254F, 255R, 259A, 261 E, 266P, 267P, 268A, 270A, 270R, 272L, 272P, 273K, 273P, 273T, 275L, 277K, 279G, 282G, 287L, 287R, 289T, 290T, 291 S, 292W, 293V, 304R, 307L, 309G, 319R, 320L, 320N, 320R, 320S, 343G, 343R, 344V, 346G, 346N, 355G, 365I, 370F, 373R, 380G, 380R, 382R, 386D, 386Q, 389G, 390R, 408A, 412A, 412H, 4121, 412R, 414R, 415S, 418S, 423D, 424V, and 468T. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 606 and one or more residue differences as compared to SEQ ID NO: 606, selected from S2A, E12L, E12M, H13I, H13N, H13Y, E15R, K16S, A18Q, T19L, P22S, P22T, K26R, E30H, T31 M / V262A, D34G, K51 R, F60L, E69A, E69L, T79Y, L82Q, Q84R, P87T, E88L, S106Q, D107G, D107H, D107R, S113E, V114P, K115S, L128F, F132G, F132R, T134E, T134G, T134L, K135T, T136L, I140C, I140L, 1140V, D143G, D143R, D143V, M146K, H150R, E152S, G160Q, G160T, A163L, A163T, H167R, F168Q, D175T, D175V, R179G, V183L, Q195G, Q195S, Q198S, S199R, S199T, S199W, N201 G, N201 Q, L215P, L215V, S217F, S217G, E218G, L219S, P220R, E223R, E223V, Y224A, Y224S, E226L, S227Y, L231 V, A233R, T235A, T235V, I236T, V243L, V243M, P254F, A255R, S259A, S261 E, A266P, V267P, T268A, K270A, K270R, K272L, K272P, D273K, D273P, D273T, M275L, P277K, N279G, Y282G, N287L, N287R, P289T, K290T, D291 S, M292W, E293V, P304R, T307L, S309G, T319R, A320L, A320N, A320R, A320S, E343G, E343R, E344V, R346G, R346N, L355G, L365I, T370F, Y373R,L380G, L380R, D382R, E386D, E386Q, S389G, Q390R, G408A, G412A, G412H, G412I, G412R, N414R, G415S, V418S, W423D, A424V, and M468T.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 606 and one or more residue differences as compared to SEQ ID NO: 606, selected from 44 / 45 / 47 / 408 / 453, 44 / 45 / 47 / 453, 44 / 45 / 47 / 453 / 454, 44 / 45 / 103 / 453 / 457, 44 / 45 / 453 / 454 / 457, 44 / 47 / 103 / 393 / 408 / 453 / 454, 44 / 47 / 103 / 453 / 454 / 457, 44 / 47 / 393 / 408 / 453 / 457, 44 / 47 / 393 / 453 / 457, 44 / 103 / 408 / 453 / 454, 45 / 47 / 103 / 393 / 453 / 454 / 457, 45 / 47 / 393 / 453 / 454, 45 / 47 / 408 / 423 / 453 / 457, 45 / 47 / 453, 45 / 103 / 393 / 453 / 454, 45 / 453 / 454, 47 / 103 / 408 / 453 / 454, 47 / 393 / 408 / 453 / 454 / 457, 47 / 393 / 453 / 454, 47 / 408 / 453 / 457, 53 / 54 / 244 / 450, 103 / 393 / 408 / 453 / 457, 244 / 450, 408 / 453 / 454, 446 / 450 / 464, and 453 / 457. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 606 and one or more residue differences as compared to SEQ ID NO: 606, selected from 44G / 45V / 47M / 408R / 453R, 44G / 45V / 47M / 453H, 44G / 45V / 47M / 453R / 454R, 44G / 47M / 393A / 453R / 457G, 44S / 45V / 103M / 453R / 457G, 44S / 45V / 453T / 454R / 457G, 44S / 47M / 103M / 393A / 408R / 453R / 454R, 44S / 47M / 103M / 453R / 454R / 457G, 44S / 47S / 393A / 408R / 453R / 457G, 44S / 103M / 408R / 453R / 454R, 45V / 47M / 393A / 453R / 454R, 45V / 47M / 408R / 423C / 453T / 457G, 45V / 47S / 103M / 393A / 453R / 454R / 457G, 45V / 47S / 453R, 45V / 103M / 393A / 453T / 454R, 45 V / 453 R / 454R, 47M / 103M / 408R / 453T / 454R, 47M / 393A / 408R / 453R / 454R / 457G, 47M / 393A / 453R / 454R, 47S / 408R / 453R / 457G, 53D / 54V / 244I / 450T, 103M / 393A / 408R / 453R / 457G, 244I / 450T, 408R / 453T / 454R, 446K / 450T / 464L, and 453R / 457G. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 606 and one or more residue differences as compared to SEQ ID NO: 606, selected from T44G / I45V / C47M / G408R / D453R, T44G / I45V / C47M / D453H, T44G / I45V / C47M / D453R / T454R, T44G / C47M / E393A / D453R / S457G, T44S / I45V / Q103M / D453R / S457G, T44S / I45V / D453T / T454R / S457G, T44S / C47M / Q103M / E393A / G408R / D453R / T454R, T44S / C47M / Q103M / D453R / T454R / S457G, T44S / C47S / E393A / G408R / D453R / S457G, T44S / Q103M / G408R / D453R / T454R, I45V / C47M / E393A / D453R / T454R, I45V / C47M / G408R / W423C / D453T / S457G, I45V / C47S / Q103M / E393A / D453R / T454R / S457G, I45V / C47S / D453R, I45V / Q103M / E393A / D453T / T454R, I45V / D453R / T454R, C47M / Q103M / G408R / D453T / T454R, C47M / E393A / G408R / D453R / T454R / S457G, C47M / E393A / D453R / T454R, C47S / G408R / D453R / S457G, M53D / Q54V / V244I / E450T, Q103M / E393A / G408R / D453R / S457G, V244I / E450T, G408R / D453T / T454R, E446K / E450T / V464L, and D453R / S457G.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 972 and one or more residue differences as compared to SEQ ID NO: 972,selected from 38 / 45 / 53, 38 / 45 / 53 / 54 / 476, 38 / 45 / 53 / 244, 38 / 45 / 53 / 470 / 472, 38 / 45 / 54 / 244, 38 / 45 / 54 / 470 / 474 / 478, 38 / 45 / 472 / 476 / 478, 38 / 45 / 476, 38 / 53 / 54, 38 / 53 / 54 / 244, 38 / 53 / 54 / 244 / 472 / 478, 38 / 53 / 472, 38 / 54 / 244 / 470 / 474 / 476 / 478, 38 / 244 / 470 / 472 / 474 / 478, 38 / 478, 45 / 53 / 54 / 244 / 470 / 474 / 476 / 478, 45 / 53 / 54 / 470 / 474 / 476, 45 / 53 / 54 / 472 / 478, 45 / 54 / 244 / 474 / 478, 53, 53 / 54 / 470 / 476 / 478, 53 / 244 / 474 / 478, 54, 54 / 244, 54 / 244 / 472 / 476, 54 / 472, 244, and 474 / 478. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 972 and one or more residue differences as compared to SEQ ID NO: 972, selected from 38F / 45I / 53D, 38F / 45I / 53D / 54S / 476S, 38F / 45I / 53L / 244I, 38F / 45I / 53L / 470N / 472R, 38F / 45I / 54V / 244T, 38F / 45I / 54V / 470N / 474S / 478G, 38F / 45I / 472R / 476S / 478G, 38F / 45I / 476S, 38F / 53D / 54H / 244I / 472R / 478G, 38F / 53D / 54S, 38F / 53D / 54S / 244I, 38F / 53D / 54S / 244I / 472R / 478G, 38F / 53L / 472R, 38F / 54S / 244I / 470N / 474S / 476S / 478G, 38F / 244I / 470R / 472R / 474S / 478G, 38F / 478G, 451 / 53 D / 54H / 244I / 470N / 474S / 476S / 478G , 451 / 53 D / 54S / 470 R / 474S / 476S , 451 / 53L / 54S / 472 R / 478G , 45I / 54S / 244T / 474S / 478G, 53D, 53D / 244T / 474S / 478G, 53L / 54S / 470R / 476S / 478G, 54H, 54S / 472R, 54V / 244I / 472R / 476S, 54V / 244T, 244T, and 474S / 478G. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 972 and one or more residue differences as compared to SEQ ID NO: 972, selected from C38F / V45I / M53D, C38F / V45I / M53D / Q54S / G476S, C38F / V45I / M53L / V244I, C38F / V45I / M53L / A470N / G472R, C38F / V45I / Q54V / V244T, C38F / V45I / Q54V / A470N / L474S / S478G, C38F / V45I / G472R / G476S / S478G, C38F / V45I / G476S, C38F / M53D / Q54H / V244I / G472R / S478G, C38F / M53D / Q54S, C38F / M53D / Q54S / V244I, C38F / M53D / Q54S / V244I / G472R / S478G, C38F / M53L / G472R, C38F / Q54S / V244I / A470N / L474S / G476S / S478G, C38 F / V244I / A470 R / G472 R / L474S / S478G , C38 F / S478G ,V45I / M53D / Q54H / V244I / A470N / L474S / G476S / S478G, V45I / M53D / Q54S / A470R / L474S / G476S, V45I / M53L / Q54S / G472R / S478G, V45I / Q54S / V244T / L474S / S478G, M53D, M53D / V244T / L474S / S478G, M53L / Q54S / A470R / G476S / S478G, Q54H, Q54S / G472R, Q54V / V244I / G472R / G476S, Q54V / V244T, V244T, and L474S / S478G.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 972 and one or more residue differences as compared to SEQ ID NO: 972, selected from 22 / 38 / 45 / 107 / 259 / 317 / 412, 22 / 38 / 45 / 259 / 317 / 412, 22 / 107, 22 / 107 / 317 / 412, 22 / 134, 22 / 134 / 220, 22 / 134 / 220 / 317, 22 / 134 / 317 / 412, 22 / 317, 38, 38 / 45 / 81 / 143 / 244, 38 / 53 / 243, 38 / 81 , 38 / 81 / 244, 38 / 107 / 134 / 220, 38 / 132, 38 / 220 / 317, 45 / 81 , 45 / 81 / 243, 45 / 81 / 243 / 343, 45 / 143 / 243 / 270 / 343, 45 / 244 / 270, 53 / 115 / 163 / 243 / 270 / 343, 53 / 143 / 243 / 270 / 343, 53 / 270, 64 / 134 / 220 / 317 / 412, 81 / 132 / 243, 107, 107 / 134 / 317, 115 / 244 / 270, 134 / 259, 143 / 243 / 270, 243, 243 / 270, 244, 244 / 270, 259, 270, and 317. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 972 and one or moreresidue differences as compared to SEQ ID NO: 972, selected from 22T / 38F / 45I / 107R / 259A / 317M / 412H, 22T / 38F / 45I / 259A / 317M / 412H, 22T / 107R, 22T / 107R / 317M / 412H, 22T / 134G, 22T / 134G / 220R / 317M, 22T / 134G / 317M / 412H, 22T / 134L / 220R, 22T / 317M, 38F, 38F / 45I / 81 T / 143R / 244I, 38F / 53D / 243L, 38F / 81T, 38F / 81 T / 244I, 38F / 107R / 134G / 220R, 38F / 132G, 38F / 220R / 317M, 451 / 81 T, 451 / 81 T / 243L, 451 / 81 T / 243L / 343R, 451 / 143R / 243L / 270R / 343R, 45I / 244I / 270R, 53D / 115R / 163L / 243L / 270R / 343R, 53D / 143R / 243L / 270R / 343R, 53D / 270R, 64R / 134L / 220R / 317M / 412H, 81T / 132G / 243L, 107R, 107R / 134L / 317M, 115R / 244I / 270R, 134G / 259A, 143R / 243L / 270R, 243L, 243L / 270R, 244I, 244I / 270R, 259A, 270R, and 317M. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93% 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 972 and one or more residue differences as compared to SEQ ID NO: 972, selected fromP22T / C38F / V45I / D107R / S259A / R317M / G412H, P22T / C38F / V45I / S259A / R317M / G412H, P22T / D107R, P22T / D107R / R317M / G412H, P22T / T134G, P22T / T134G / P220R / R317M, P22T / T134G / R317M / G412H, P22T / T134L / P220R, P22T / R317M, C38F, C38F / V45I / V81T / D143R / V244I, C38F / M53D / V243L, C38F / V81T, C38F / V81T / V244I, C38F / D107R / T134G / P220R, C38F / F132G, C38F / P220R / R317M, V45I / V81T, V45I / V81T / V243L, V45I / V81T / V243L / E343R, V45I / D143R / V243L / K270R / E343R, V45I / V244I / K270R, M53D / K115R / A163L / V243L / K270R / E343R, M53D / D143R / V243L / K270R / E343R, M53D / K270R, P64R / T134L / P220R / R317M / G412H, V81T / F132G / V243L, D107R, D107R / T134L / R317M, K115R / V244I / K270R, T134G / S259A, D143R / V243L / K270R, V243L, V243L / K270R, V244I, V244I / K270R, S259A, K270R, and R317M.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 972 and one or more residue differences as compared to SEQ ID NO: 972, selected from 22 / 38 / 45 / 107 / 259 / 317 / 412, 22 / 38 / 45 / 259 / 317 / 412, 22 / 38 / 64 / 134 / 220, 22 / 45 / 220 / 259, 22 / 64 / 107 / 259 / 317 / 412, 22 / 64 / 134 / 220, 22 / 107, 22 / 134, 22 / 134 / 317, 22 / 134 / 317 / 412, 22 / 220, 22 / 220 / 412 / 415, 22 / 259 / 317, 22 / 259 / 317 / 412, 22 / 317, 38, 38 / 45 / 81 / 143 / 244, 38 / 45 / 143 / 270, 38 / 53 / 243, 38 / 81 , 38 / 81 / 143 / 243 / 270, 38 / 81 / 183 / 243 / 270, 38 / 81 / 243, 38 / 81 / 244, 38 / 107 / 134 / 220, 38 / 132, 38 / 220 / 317, 38 / 270 / 343, 45 / 81 , 45 / 81 / 132 / 243 / 270, 45 / 81 / 243, 45 / 143 / 243 / 270 / 343, 45 / 244 / 270, 53 / 143 / 243 / 270 / 343, 53 / 244 / 270, 53 / 270, 64, 64 / 107 / 134, 64 / 107 / 317, 64 / 134, 64 / 134 / 220 / 317 / 412, 81 / 132, 81 / 132 / 243, 107, 107 / 134 / 317, 107 / 412, 115 / 244 / 270, 143 / 243 / 270, 220 / 259, 243, 243 / 270, 244 / 270, 255, 259, and 270. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 972 and one or more residue differences as compared to SEQ ID NO: 972, selected from 22T / 38F / 45I / 107R / 259A / 317M / 412H, 22T / 38F / 45I / 259A / 317M / 412H, 22T / 38F / 64R / 134L / 220R, 22T / 45I / 220R / 259A, 22T / 64R / 107R / 259A / 317M / 412H, 22T / 64R / 134L / 220R, 22T / 107R, 22T / 134G, 22T / 134G / 317M / 412H, 22T / 134L / 317M, 22T / 134L / 317M / 412H, 22T / 220R, 22T / 220R / 412H / 415D, 22T / 259A / 317M, 22T / 259A / 317M / 412H, 22T / 317M, 38F, 38F / 45I / 81 T / 143R / 244I, 38F / 45I / 143R / 270R, 38F / 53D / 243L, 38F / 81T, 38F / 81T / 143R / 243L / 270R, 38F / 81T / 183L / 243L / 270R, 38F / 81T / 243L,38F / 81 T / 244I, 38F / 107R / 134G / 220R, 38F / 132G, 38F / 220R / 317M, 38F / 270R / 343R, 451 / 81 T, 451 / 81 T / 132G / 243L / 270R, 451 / 81 T / 243L, 451 / 143R / 243L / 270R / 343R, 45I / 244I / 270R, 53D / 143R / 243L / 270R / 343R, 53D / 244I / 270R, 53D / 270R, 64R, 64R / 107R / 134L, 64R / 107R / 317M, 64R / 134L, 64R / 134L / 220R / 317M / 412H, 81T / 132G, 81 T / 132G / 243L, 107R, 107R / 134L / 317M, 107R / 412H, 115R / 244I / 270R, 143R / 243L / 270R, 220R / 259A, 243L, 243L / 270R, 244I / 270R, 255R, 259A, and 270R. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 972 and one or more residue differences as compared to SEQ ID NO: 972, selected fromP22T / C38F / V45I / D107R / S259A / R317M / G412H, P22T / C38F / V45I / S259A / R317M / G412H, P22T / C38F / P64R / T134L / P220R, P22T / V45I / P220R / S259A, P22T / P64R / D107R / S259A / R317M / G412H, P22T / P64R / T134L / P220R, P22T / D107R, P22T / T134G, P22T / T134G / R317M / G412H, P22T / T134L / R317M, P22T / T134L / R317M / G412H, P22T / P220R, P22T / P220R / G412H / G415D, P22T / S259A / R317M, P22T / S259A / R317M / G412H, P22T / R317M, C38F, C38F / V45I / V81T / D143R / V244I, C38F / V45I / D143R / K270R, C38F / M53D / V243L, C38F / V81T, C38F / V81T / D143R / V243L / K270R, C38F / V81T / V183L / V243L / K270R, C38F / V81T / V243L, C38F / V81T / V244I, C38F / D107R / T134G / P220R, C38F / F132G, C38F / P220R / R317M, C38F / K270R / E343R, V45I / V81T, V45I / V81T / F132G / V243L / K270R, V45I / V81 T / V243L, V45I / D143R / V243L / K270R / E343R, V45I / V244I / K270R, M53D / D143R / V243L / K270R / E343R, M53D / V244I / K270R, M53D / K270R, P64R, P64R / D107R / T134L, P64R / D107R / R317M, P64R / T134L, P64R / T134L / P220R / R317M / G412H, V81T / F132G, V81T / F132G / V243L, D107R, D107R / T134L / R317M, D107R / G412H, K115R / V244I / K270R, D143R / V243L / K270R, P220R / S259A, V243L, V243L / K270R, V244I / K270R, A255R, S259A, and K270R.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 1082 and one or more residue differences as compared to SEQ ID NO: 1082, selected from 143, 143 / 175 / 272 / 273 / 415, 152 / 168 / 216 / 237, 152 / 168 / 423, 160, 160 / 201 / 272, 160 / 201 / 273 / 415, 160 / 226, 160 / 244 / 272 / 273 / 415, 160 / 415, 168, 168 / 187 / 216 / 255 / 423, 168 / 216 / 237, 168 / 255 / 423, 168 / 290 / 423, 187, 187 / 423, 199 / 201 / 226, 199 / 201 / 415, 201 , 216, 216 / 423, 226 / 244 / 415, 226 / 415, 237, 237 / 290, 244, 255, 272, 290, 290 / 423, 415, and 423. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 1082 and one or more residue differences as compared to SEQ ID NO: 1082, selected from 143R, 143R / 175T / 272W / 273P / 415S, 152S / 168Q / 216E / 237S, 152S / 168Q / 423D, 160Q, 160Q / 201 G / 272W, 160Q / 201 G / 273P / 415S, 160Q / 226R, 160Q / 244I / 272W / 273P / 415S, 160Q / 415S, 168Q, 168Q / 187G / 216E / 255R / 423D, 168Q / 216E / 237S, 168Q / 255R / 423D, 168Q / 290R / 423D, 187G, 187G / 423D, 199W / 201 G / 226R, 199W / 201 G / 415S, 201 G, 216E, 216E / 423D, 226R / 244I / 415S, 226R / 415S, 237S, 237S / 290R, 244I, 255R, 272W, 290R, 290R / 423D, 415S, and 423D. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%,93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 1082 and one or more residue differences as compared to SEQ ID NO: 1082, selected from D143R, D143R / D175T / K272W / D273P / G415S, E152S / F168Q / D216E / D237S, E152S / F168Q / W423D, G160Q, G160Q / N201 G / K272W, G160Q / N201 G / D273P / G415S, G160Q / E226R, G160Q / V244I / K272W / D273P / G415S, G160Q / G415S, F168Q, F168Q / E187G / D216E / A255R / W423D, F168Q / D216E / D237S, F168Q / A255R / W423D, F168Q / K290R / W423D, E187G, E187G / W423D, S199W / N201 G / E226R, S199W / N201 G / G415S, N201 G, D216E, D216E / W423D, E226R / V244I / G415S, E226R / G415S, D237S, D237S / K290R, V244I, A255R, K272W, K290R, K290R / W423D, G415S, and W423D.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 1082 and one or more residue differences as compared to SEQ ID NO: 1082, selected from 143, 143 / 160 / 226 / 415, 143 / 175 / 272 / 273 / 415, 152 / 168 / 216 / 237, 152 / 168 / 423, 160, 160 / 201 / 272, 160 / 201 / 273 / 415, 160 / 226, 160 / 244 / 272 / 273 / 415, 160 / 415, 168, 168 / 187 / 216 / 255 / 423, 168 / 216 / 237, 168 / 216 / 237 / 255 / 343, 168 / 255 / 423, 168 / 290 / 423, 187, 199 / 201 / 226, 199 / 201 / 415, 201 , 216, 216 / 423, 226 / 244 / 415, 226 / 415, 237, 237 / 290, 244, 255, 255 / 343, 272, 290, 290 / 423, 415, and 423. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 1082 and one or more residue differences as compared to SEQ ID NO: 1082, selected from 143R, 143R / 160T / 226R / 415S, 143R / 175T / 272W / 273P / 415S, 152S / 168Q / 216E / 237S, 152S / 168Q / 423D, 160Q, 160Q / 201 G / 272W, 160Q / 201 G / 273P / 415S, 160Q / 226R, 160Q / 244I / 272W / 273P / 415S, 160Q / 415S, 168Q, 168Q / 187G / 216E / 255R / 423D, 168Q / 216E / 237S, 168Q / 216E / 237S / 255R / 343R, 168Q / 255R / 423D, 168Q / 290R / 423D, 187G, 199W / 201 G / 226R, 199W / 201 G / 415S, 201 G, 216E, 216E / 423D, 226R / 244I / 415S, 226R / 415S, 237S, 237S / 290R, 244I, 255R, 255R / 343R, 272W, 290R, 290R / 423D, 415S, and 423D. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 1082 and one or more residue differences as compared to SEQ ID NO: 1082, selected from D143R, D143R / G160T / E226R / G415S, D143R / D175T / K272W / D273P / G415S, E152S / F168Q / D216E / D237S, E152S / F168Q / W423D, G160Q, G160Q / N201 G / K272W, G160Q / N201 G / D273P / G415S, G160Q / E226R, G160Q / V244I / K272W / D273P / G415S, G160Q / G415S, F168Q, F168Q / E187G / D216E / A255R / W423D, F168Q / D216E / D237S, F168Q / D216E / D237S / A255R / E343R, F168Q / A255R / W423D, F168Q / K290R / W423D, E187G, S199W / N201 G / E226R, S199W / N201 G / G415S, N201 G, D216E, D216E / W423D, E226R / V244I / G415S, E226R / G415S, D237S, D237S / K290R, V244I, A255R, A255R / E343R, K272W, K290R, K290R / W423D, G415S, and W423D.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to referencesequence SEQ ID NO: 1082 and one or more residue differences as compared to SEQ ID NO: 1082, selected from 72, 90, 91 , 92, 93, 95, 122, 129, 141 , 142, 143, 144, 145, 150, 151 , 151 / 155, 152, 154, 155, 157, 160, 161 , 161 / 313, 164, 165, 168, 180, 181 , 184, 193, 197, 204, 205, 205 / 472, 206, 299, and 320. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 1082 and one or more residue differences as compared to SEQ ID NO: 1082, selected from 72M, 72Q, 90F, 911, 91 V, 92A, 92L, 92V, 93A, 93H, 93S, 95G, 95S, 122F, 122L, 122Q, 122V, 129F, 1291, 141 Y, 142L, 143A, 143K, 144F, 1441, 144V, 144Y, 145F, 145L, 150D, 150E, 150G, 150R, 150S, 150T, 150V, 150W, 151 A, 151Q, 151 R, 151T / 155T, 152D, 152H, 152L, 152Q, 152R, 152T, 152V, 152Y, 154E, 154K, 154L, 154P, 154Q, 154S, 154T, 154V, 154Y, 155E, 155G, 155H, 155Q, 155R, 155S, 155T, 157R, 1601, 160Q, 160V, 160W, 161 C, 161 C / 313C, 161 R, 161 V, 164Q, 164R, 164S, 165G, 165S, 168E, 168K, 168L, 168N, 168R, 168V, 180C, 180L, 180T, 181 L, 181 V, 184A, 184G, 184H, 184S, 193H, 193L, 193M, 193V, 197H, 197R, 197Y, 204L, 204M, 204V, 204W, 204Y, 205I, 205L, 205L / 472D, 206L, 299F, 299I, 320C, 320H, 320K, 320L, 320Q, 320R, 320S, and 320Y. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 1082 and one or more residue differences as compared to SEQ ID NO: 1082, selected from R72M, R72Q, L90F, A911, A91 V, I92A, I92L, I92V, D93A, D93H, D93S, T95G, T95S, W122F, W122L, W122Q, W122V, L129F, L129I, H141Y, Q142L, D143A, D143K, W144F, W144I, W144V, W144Y, W145F, W145L, H150D, H150E, H150G, H150R, H150S, H150T, H150V, H150W, P151 A, P151 Q, P151 R, P151T / D155T, E152D, E152H, E152L, E152Q, E152R, E152T, E152V, E152Y, A154E, A154K, A154L, A154P, A154Q, A154S, A154T, A154V, A154Y, D155E, D155G, D155H, D155Q, D155R, D155S, D155T, K157R, G160I, G160Q, G160V, G160W, K161 C, K161 C / R313C, K161 R, K161 V, D164Q, D164R, D164S, A165G, A165S, F168E, F168K, F168L, F168N, F168R, F168V, V180C, V180L, V180T, 1181 L, 1181 V, C184A, C184G, C184H, C184S, Y193H, Y193L, Y193M, Y193V, K197H, K197R, K197Y, F204L, F204M, F204V, F204W, F204Y, V205I, V205L, V205L / G472D, V206L, L299F, L299I, A320C, A320H, A320K, A320L, A320Q, A320R, A320S, and A320Y.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 1082 and one or more residue differences as compared to SEQ ID NO: 1082, selected from 90, 92, 122, 129, 141 , 143, 144, 145, 150, 151 , 151 / 155, 152, 154, 155, 155 / 265, 157, 160, 161 , 164, 165, 168, 180, 181 , 184, 193, 197, 204, 205, 205 / 472, 206, 299, and 320. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 1082 and one or more residue differences as compared to SEQ ID NO: 1082, selected from 90F, 92A, 92L, 92P, 92V, 122F, 122L, 129F, 1291, 141 Y, 143A, 143K, 144F, 144Y, 145F, 145L, 150D, 150E, 150G, 150R, 150S, 150V, 150W, 151 A, 151 K, 151Q, 151 R, 151T / 155T, 152D, 152H, 152L, 152Q, 152R, 152T, 152V, 152Y, 154E, 154K, 154L,154Q, 154S, 154T, 154V, 154Y, 155E, 155G, 155H, 155Q, 155Q / 265V, 155R, 155S, 155T, 157R, 1601, 160Q, 160V, 160W, 161 C, 161 R, 161 V, 164Q, 164R, 164S, 165G, 165S, 168E, 168K, 168L, 168N, 168R, 168V, 180A, 180C, 180L, 180T, 181 L, 181 V, 184A, 184G, 184S, 193H, 193L, 193M, 193V, 197H, 197R, 204L, 204M, 204V, 204W, 204Y, 205I, 205L, 205L / 472D, 206L, 299F, 299I, 320C, 320H, 320K, 320L, 320Q, 320R, 320S, and 320Y. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 1082 and one or more residue differences as compared to SEQ ID NO: 1082, selected from L90F, I92A, I92L, I92P, I92V, W122F, W122L, L129F, L129I, H141 Y, D143A, D143K, W144F, W144Y, W145F, W145L, H150D, H150E, H150G, H150R, H150S, H150V, H150W, P151 A, P151 K, P151 Q, P151 R, P151 T / D155T, E152D, E152H, E152L, E152Q, E152R, E152T, E152V, E152Y, A154E, A154K, A154L, A154Q, A154S, A154T, A154V, A154Y, D155E, D155G, D155H, D155Q, D155Q / A265V, D155R, D155S, D155T, K157R, G160I, G160Q, G160V, G160W, K161 C, K161 R, K161 V, D164Q, D164R, D164S, A165G, A165S, F168E, F168K, F168L, F168N, F168R, F168V, V180A, V180C, V180L, V180T, 1181 L, 1181 V, C184A, C184G, C184S, Y193H, Y193L, Y193M, Y193V, K197H, K197R, F204L, F204M, F204V, F204W, F204Y, V205I, V205L, V205L / G472D, V206L, L299F, L299I, A320C, A320H, A320K, A320L, A320Q, A320R, A320S, and A320Y.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 1218 and one or more residue differences as compared to SEQ ID NO: 1218, selected from 129, 129 / 160, 129 / 160 / 164, 129 / 160 / 184, 129 / 164, 129 / 164 / 184, 129 / 205, 146 / 152 / 244, 150 / 155 / 226, 150 / 155 / 244, 152, 154, 154 / 160 / 184, 154 / 164, 154 / 205, 155, 160, 160 / 164, 160 / 164 / 184, 160 / 184, 164, 164 / 204, 164 / 205, 204, 205, 226, 226 / 244, and 244. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 1218 and one or more residue differences as compared to SEQ ID NO: 1218, selected from 1291, 1291 / 1601, 1291 / 1601 / 164Q, 1291 / 1601 / 184A, 1291 / 164Q, 1291 / 164Q / 184A, 1291 / 2051, 146L / 152T / 244I, 150D / 155T / 226R, 150D / 155T / 244I, 152T, 154K, 154K / 1601 / 184A, 154K / 164Q, 154K / 205I, 155T, 1601, 1601 / 164Q, 1601 / 164Q / 184A, 1601 / 184A, 164Q, 164Q / 204L, 164Q / 205I, 204L, 2051, 226R, 226R / 244I, and 2441. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 1218 and one or more residue differences as compared to SEQ ID NO: 1218, selected from L129I, L129I / G160I, L129I / G160I / D164Q, L129I / G160I / C184A, L129I / D164Q, L129I / D164Q / C184A, L129I / V205I, M146L / E152T / V244I, H150D / D155T / E226R, H150D / D155T / V244I, E152T, A154K, A154K / G160I / C184A, A154K / D164Q, A154K / V205I, D155T, G160I, G160I / D164Q, G160I / D164Q / C184A, G160I / C184A, D164Q, D164Q / F204L, D164Q / V205I, F204L, V205I, E226R, E226R / V244I, and V244I.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 1218 and one or more residue differences as compared to SEQ ID NO: 1218, selected from 129, 129 / 143, 129 / 154 / 164, 129 / 154 / 164 / 205, 129 / 154 / 205, 129 / 160, 129 / 160 / 164, 129 / 160 / 205, 129 / 164 / 205, 143, 143 / 150 / 155 / 226 / 244, 143 / 150 / 155 / 244, 143 / 150 / 226, 143 / 150 / 226 / 244 / 343, 143 / 150 / 226 / 343, 143 / 150 / 244, 143 / 150 / 343, 143 / 152, 143 / 152 / 226, 143 / 152 / 226 / 244, 143 / 152 / 226 / 343, 143 / 152 / 343, 143 / 155, 143 / 155 / 226 / 244, 143 / 155 / 226 / 244 / 343, 143 / 155 / 343, 143 / 226, 143 / 226 / 244 / 343, 143 / 226 / 343, 143 / 244, 143 / 244 / 343, 143 / 343, 146 / 152 / 244, 150 / 155 / 226, 150 / 155 / 244, 150 / 155 / 244 / 343, 150 / 226 / 244 / 343, 152, 152 / 174 / 244 / 343, 152 / 226, 152 / 226 / 343, 152 / 343, 154, 154 / 160 / 184, 154 / 164, 154 / 164 / 204, 154 / 164 / 204 / 205, 154 / 205, 155, 160, 160 / 164, 160 / 164 / 184, 164, 205, 226, 226 / 244, and 244. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 1218 and one or more residue differences as compared to SEQ ID NO: 1218, selected from 1291, 1291 / 143H, 1291 / 154K / 164Q, 1291 / 154K / 164Q / 205I, 1291 / 154K / 205I, 1291 / 1601, 1291 / 1601 / 164Q, 1291 / 1601 / 2051, 1291 / 164Q / 2051, 143R, 143R / 150D / 155T / 226R / 244I, 143R / 150D / 155T / 244I, 143R / 150D / 226R, 143R / 150D / 226R / 244I / 343R, 143R / 150D / 226R / 343R, 143R / 150D / 244I, 143R / 150D / 343R, 143R / 152T, 143R / 152T / 226R, 143R / 152T / 226R / 244I, 143R / 152T / 226R / 343R, 143R / 152T / 343R, 143R / 155T, 143R / 155T / 226R / 244I, 143R / 155T / 226R / 244I / 343R, 143R / 155T / 343R, 143R / 226R, 143R / 226R / 244I / 343R, 143R / 226R / 343R, 143R / 244I, 143R / 244I / 343R, 143R / 343R, 146L / 152T / 2441 , 150D / 155T / 226R, 150D / 155T / 244I, 150D / 155T / 244I / 343R, 150D / 226R / 244I / 343R, 152T, 152T / 174R / 244I / 343R, 152T / 226R, 152T / 226R / 343R, 152T / 343R, 154K, 154K / 1601 / 184A, 154K / 164Q, 154K / 164Q / 204L, 154K / 164Q / 204L / 205I, 154K / 205I, 155T, 1601, 1601 / 164Q, 1601 / 164Q / 184A, 164Q, 205I, 226R, 226R / 244I, and 244I. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 1218 and one or more residue differences as compared to SEQ ID NO: 1218, selected from L129I, L129I / D143H, L129I / A154K / D164Q, L129I / A154K / D164Q / V205I, L129I / A154K / V205I , L129I / G160I, L129I / G160I / D164Q, L129I / G160I / V205I, L129I / D164Q / V205I, D143R, D143R / H150D / D155T / E226R / V244I, D143R / H150D / D155T / V244I, D143R / H150D / E226R, D143R / H150D / E226R / V244I / E343R, D143R / H150D / E226R / E343R, D143R / H150D / V244I, D143R / H150D / E343R, D143R / E152T, D143R / E152T / E226R, D143R / E152T / E226R / V244I, D143R / E152T / E226R / E343R, D143R / E152T / E343R, D143R / D155T, D143R / D155T / E226R / V244I, D143R / D155T / E226R / V244I / E343R, D143R / D155T / E343R, D143R / E226R, D143R / E226R / V244I / E343R, D143R / E226R / E343R, D143R / V244I, D143R / V244I / E343R, D143R / E343R, M146L / E152T / V244I, H150D / D155T / E226R, H150D / D155T / V244I, H150D / D155T / V244I / E343R, H150D / E226R / V244I / E343R, E152T, E152T / G174R / V244I / E343R, E152T / E226R, E152T / E226R / E343R, E152T / E343R, A154K,A154K / G160I / C184A, A154K / D164Q, A154K / D164Q / F204L, A154K / D164Q / F204L / V205I, A154K / V205I, D155T, G160I, G160I / D164Q, G160I / D164Q / C184A, D164Q, V205I, E226R, E226R / V244I, and V244I.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 1580 and one or more residue differences as compared to SEQ ID NO: 1580, selected from 98 / 129 / 157 / 168, 129 / 157 / 168, 151 , 151 / 152 / 255, 151 / 152 / 273, 151 / 255, 151 / 343, 157, 157 / 164, 157 / 164 / 168, 157 / 168, 164, 164 / 255, 168, 183, 255, 255 / 343, and 343. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 1580 and one or more residue differences as compared to SEQ ID NO: 1580, selected from 981 / 1291 / 157R / 168K, 1291 / 157R / 168K, 151 Q, 151 Q / 152T / 255R, 151 Q / 152T / 273Y, 151 Q / 255R, 151 R / 343R, 157R, 157R / 164D, 157R / 164R, 157R / 164R / 168K, 157R / 168K, 164D, 164D / 255R, 168K, 183I, 255R, 255R / 343R, and 343R. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 1580 and one or more residue differences as compared to SEQ ID NO: 1580, selected from L98I / L129I / K157R / Q168K, L129I / K157R / Q168K, P151 Q, P151 Q / E152T / A255R, P151 Q / E152T / D273Y, P151 Q / A255R, P151 R / E343R, K157R, K157R / Q164D, K157R / Q164R, K157R / Q164R / Q168K, K157R / Q168K, Q164D, Q164D / A255R, Q168K, V183I, A255R, A255R / E343R, and E343R.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 1580 and one or more residue differences as compared to SEQ ID NO: 1580, selected from 129, 129 / 157, 129 / 157 / 168, 129 / 164, 129 / 164 / 168 / 183, 129 / 164 / 183, 143, 143 / 151 , 143 / 151 / 152, 143 / 151 / 164, 143 / 164, 143 / 164 / 255, 143 / 164 / 343, 143 / 255, 143 / 343, 151 , 151 / 152, 151 / 152 / 164 / 343, 151 / 152 / 343, 151 / 255, 151 / 255 / 343, 151 / 343, 152, 152 / 255, 157, 157 / 164 / 168, 157 / 168, 164, 164 / 168, 164 / 255, 164 / 447, 168, 183, 255, 255 / 343, 298 / 343, and 343. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 1580 and one or more residue differences as compared to SEQ ID NO: 1580, selected from 1291, 1291 / 157R, 1291 / 157R / 168K, 1291 / 164R, 1291 / 164R / 168K / 183M, 1291 / 164R / 183I, 143R, 143R / 151 Q, 143R / 151 R, 143R / 151 R / 152T, 143R / 151 R / 164D, 143R / 164D, 143R / 164D / 255R, 143 R / 164 D / 343 R, 143R / 255R, 143R / 343R, 151 R, 151 R / 152T, 151 R / 152T / 164D / 343R, 151 R / 152T / 343R, 151 R / 255R, 151 R / 255R / 343R, 151 R / 343R, 152T, 152T / 255R, 157R, 157R / 164R / 168K, 157R / 168K, 164D, 164D / 255R, 164R, 164R / 168K, 164R / 447T, 168K, 1831, 255R, 255R / 343R, 298I / 343R, and 343R. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to referencesequence SEQ ID NO: 1580 and one or more residue differences as compared to SEQ ID NO: 1580, selected from L129I, L129I / K157R, L129I / K157R / Q168K, L129I / Q164R, L129I / Q164R / Q168K / V183M, L129I / Q164R / V183I, D143R, D143R / P151 Q, D143R / P151 R, D143R / P151 R / E152T,D143R / P151 R / Q164D, D143R / Q164D, D143R / Q164D / A255R, D143R / Q164D / E343R, D143R / A255R, D143R / E343R, P151 R, P151 R / E152T, P151 R / E152T / Q164D / E343R, P151 R / E152T / E343R, P151 R / A255R, P151 R / A255R / E343R, P151 R / E343R, E152T, E152T / A255R, K157R,K157R / Q164R / Q168K, K157R / Q168K, Q164D, Q164D / A255R, Q164R, Q164R / Q168K, Q164R / A447T, Q168K, V183I, A255R, A255R / E343R, V298I / E343R, and E343R.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 1580 and one or more residue differences as compared to SEQ ID NO: 1580, selected from 69, 69 / 164, 72, 73, 91 , 92 / 164, 95, 96, 97, 98, 1 10, 1 12, 1 13, 121 , 121 / 164, 122, 122 / 164, 124, 126, 127, 139, 140, 141 , 144, 144 / 164, 145, 146, 147, 148, 150, 150 / 164, 151 , 151 / 164, 153, 153 / 164, 155, 156, 157, 159, 160, 161 , 163, 164, 164 / 172, 164 / 173, 164 / 182, 164 / 188, 164 / 191 , 164 / 192, 164 / 195, 164 / 197, 164 / 199, 164 / 202, 164 / 205 / 249, 164 / 209, 165, 166, 167, 169, 170, 171 , 172, 173, 177, 184, 187, 188, 190, 191 , 192, 193, 194, 195, 197, 197 / 266, 198, 199, 200, 202, 203, 204, 205, 208, 209, 210, 321 , 324, 331 , and 349. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 1580 and one or more residue differences as compared to SEQ ID NO: 1580, selected from 69Q, 69V / 164D, 72L, 72Q, 73Q, 73W, 91 C, 91 L, 91 V, 92L / 164D, 95G, 95S, 95V, 96G, 97T, 98V, 1 1 OR, 1 10T, 1 10V, 1 12A, 1 12H, 1 13V, 121 A, 121 L / 164D, 121 Q, 122C, 122F, 122L, 122V, 122V / 164D, 124L, 124M, 126A, 126S, 127A, 1271, 1391, 140L, 140V, 141 T, 144F, 144L, 144L / 164D, 144V, 145F, 145G, 145K, 145L, 145Q, 145R, 145S, 145T, 146A, 146F, 146G, 146K, 146Q, 146R, 146V, 147W, 148A, 148E, 148F, 148G, 148R, 148V, 150A, 150A / 164D, 150G, 150L, 150L / 164D, 150R, 150R / 164D, 150S / 164D, 150T, 150V, 150W, 151 A, 151 C, 151 E, 151 G, 151 1, 151 K, 151 L, 151 L / 164D, 151 M, 151 N, 151 Q, 151 S, 151 T, 151 V, 151 W, 151 Y, 151 Y / 164D, 153G, 153G / 164D, 153N, 153P, 153R, 155F, 156C, 156G, 156L, 156Y, 157A, 157C, 157H, 157P, 157Q, 157R, 157S, 157V, 159A, 159G, 159P, 159Q, 159T, 160A, 160L, 160Q, 160S, 160V, 160Y, 161 A, 161 C, 161 F, 161 L, 161 M, 161 T, 163D, 163G, 163K, 163N, 163Q, 163W, 164D, 164D / 172M, 164D / 173M, 164D / 182W, 164D / 188G, 164D / 191 R, 164D / 192E, 164D / 195R, 164D / 197A, 164D / 199G, 164D / 199L, 164D / 202A, 164D / 205L / 249E, 164D / 209R, 165H, 165T, 165V, 166M, 166R, 166V, 166W, 167L, 167R, 167V, 1691, 169S, 169T, 169Y, 170L, 171 1, 171 R, 171 S, 1721, 172S, 172T, 172V, 1731, 177G, 177R, 177S, 177T, 177V, 177W, 177Y, 184E, 184G, 184Q, 184S, 184T, 187G, 188G, 190D, 190G, 190M, 190W, 191 G, 191 L, 191 W, 192A, 192L, 193W, 194C, 194R, 194W, 195G, 195H, 195W, 197A, 197L / 266V, 197M, 197S, 198A, 198G, 1981, 198K, 198L, 198N, 198R, 199G, 199L, 199W, 199Y, 200A, 200F, 200L, 200R, 200V, 200Y, 202F, 202I, 202L, 202S, 202V, 203Q, 204I, 204T, 204V, 205T, 208C, 209A, 209G, 209V, 2101, 21 OP, 21 OR, 210T, 210V, 321 A, 321 S, 324M, 331 L, 349K, and 349L. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%,97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 1580 and one or more residue differences as compared to SEQ ID NO: 1580, selected from E69Q, E69V / Q164D, R72L, R72Q, K73Q, K73W, A91 C, A91 L, A91 V, I92L / Q164D, T95G, T95S, T95V, T96G, S97T, L98V, K1 10R, K1 10T, K1 10V, G1 12A, G1 12H, S1 13V, W121 A, W121 L / Q164D, W121 Q, W122C, W122F, W122L, W122V, W122V / Q164D, H124L, H124M, T126A, T126S, L127A, L127I, L139I, I140L, 1140V, H141 T, W144F, W144L, W144L / Q164D, W144V, W145F, W145G, W145K, W145L, W145Q, W145R, W145S, W145T, M146A, M146F, M146G, M146K, M146Q, M146R, M146V, R147W, P148A, P148E, P148F, P148G, P148R, P148V, H150A, H150A / Q164D, H150G, H150L, H150L / Q164D, H150R, H150R / Q164D, H150S / Q164D, H150T, H150V, H150W, P151 A, P151 C, P151 E, P151 G, P151 1, P151 K, P151 L, P151 L / Q164D, P151 M, P151 N, P151 Q, P151 S, P151 T, P151 V, P151 W, P151 Y, P151 Y / Q164D, D153G, D153G / Q164D, D153N, D153P, D153R, D155F, E156C, E156G, E156L, E156Y, K157A, K157C, K157H, K157P, K157Q, K157R, K157S, K157V, S159A, S159G, S159P, S159Q, S159T, G160A, G160L, G160Q, G160S, G160V, G160Y, K161 A, K161 C, K161 F, K161 L, K161 M, K161 T, A163D, A163G, A163K, A163N, A163Q, A163W, Q164D, Q164D / R172M, Q164D / L173M, Q164D / S182W, Q164D / A188G, Q164D / Q191 R, Q164D / S192E, Q164D / Q195R, Q164D / K197A, Q164D / S199G, Q164D / S199L, Q164D / E202A, Q164D / V205L / K249E, Q164D / K209R, A165H, A165T, A165V, S166M, S166R, S166V, S166W, H167L, H167R, H167V, C169I, C169S, C169T, C169Y, R170L, Q171 1, Q171 R, Q171 S, R172I, R172S, R172T, R172V, L173I, M177G, M177R, M177S, M177T, M177V, M177W, M177Y, C184E, C184G, C184Q, C184S, C184T, E187G, A188G, I190D, I190G, I190M, I190W, Q191 G, 0191 L, Q191 W, S192A, S192L, Y193W, L194C, L194R, L194W, Q195G, Q195H, Q195W, K197A, K197L / A266V, K197M, K197S, Q198A, Q198G, Q198I, Q198K, Q198L, Q198N, Q198R, S199G, S199L, S199W, S199Y, H200A, H200F, H200L, H200R, H200V, H200Y, E202F, E202I, E202L, E202S, E202V, R203Q, F204I, F204T, F204V, V205T, A208C, K209A, K209G, K209V, H210I, H210P, H210R, H210T, H210V, R321 A, R321 S, V324M, W331 L, E349K, and E349L.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 1580 and one or more residue differences as compared to SEQ ID NO: 1580, selected from 69, 69 / 164, 72, 73, 91 , 92 / 164, 95, 97, 98, 121 / 164, 126, 127, 139, 140, 142, 144, 144 / 164, 145, 146, 146 / 164, 148, 150, 150 / 164, 151 , 153, 153 / 164, 155, 156, 157, 160, 163, 164 / 173, 164 / 188, 164 / 191 , 164 / 192, 164 / 195, 164 / 199, 164 / 205 / 249, 164 / 209, 165, 166, 167, 169, 171 , 172, 173, 177, 182, 187, 188, 190, 191 , 192, 194, 195, 197 / 266, 198, 199, 200, 202, 203, 204, 205, 208, 209, 210, 324, 331 , and 349. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 1580 and one or more residue differences as compared to SEQ ID NO: 1580, selected from 69Q, 69V, 69V / 164D, 72L, 72Q, 73Q, 73W, 91 C, 91 V, 92L / 164D, 95S, 97T, 98M, 98V, 121 L / 164D, 126A, 126S, 1271, 1391, 140L, 140V, 142C, 144F, 144L / 164D, 144V, 144Y, 145F, 145K, 145L, 146A, 146F, 146K, 146L / 164D, 146Q, 146R, 146V, 148R, 150A, 150A / 164D, 150G, 150L, 150L / 164D, 150R, 150R / 164D, 150S / 164D, 150T, 150V, 150W, 151 C, 151 E, 151 K, 151 M, 151 N, 151 Q, 151 S, 151 V, 151 W, 151 Y, 153G, 153G / 164D,153N, 153R, 155C, 155F, 155L, 155S, 155T, 156C, 156G, 156L, 156R, 156S, 156T, 156Y, 157A, 157C, 157P, 157Q, 157S, 157V, 160A, 160L, 160Q, 160S, 160V, 160Y, 163D, 163G, 163K, 163N, 163Q, 163W, 164D / 173M, 164D / 188G, 164D / 191 R, 164D / 192E, 164D / 195R, 164D / 199G, 164D / 199L, 164D / 205L / 249E, 164D / 209R, 165G, 165T, 166M, 166V, 167L, 167R, 167V, 1691, 169S, 169T, 1711, 171 R, 171 S, 171 V, 1721, 172T, 172V, 1731, 177G, 177R, 177S, 177T, 177V, 177W, 177Y, 182L, 187G, 188G, 190M, 191 F, 191 G, 192A, 192L, 194W, 195G, 195H, 195M, 195W, 195Y, 197L / 266V, 198A, 198L, 198N, 198R, 199F, 199G, 199L, 199W, 199Y, 200A, 200F, 200L, 200R, 200S, 200V, 200Y, 202L, 203Q, 204I, 204T, 204V, 205T, 208S, 209V, 210T, 210V, 324M, 331 L, and 349K. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 1580 and one or more residue differences as compared to SEQ ID NO: 1580, selected from E69Q, E69V, E69V / Q164D, R72L, R72Q, K73Q, K73W, A91 C, A91 V, I92L / Q164D, T95S, S97T, L98M, L98V, W121 L / Q164D, T126A, T126S, L127I, L139I, I140L, 1140V, Q142C, W144F, W144L / Q164D, W144V, W144Y, W145F, W145K, W145L, M146A, M146F, M146K, M146L / Q164D, M146Q, M146R, M146V, P148R, H150A, H150A / Q164D, H150G, H150L, H150L / Q164D, H150R, H150R / Q164D, H150S / Q164D, H150T, H150V, H150W, P151 C, P151 E, P151 K, P151 M, P151 N, P151 Q, P151 S, P151 V, P151W, P151Y, D153G, D153G / Q164D, D153N, D153R, D155C, D155F, D155L, D155S, D155T, E156C, E156G, E156L, E156R, E156S, E156T, E156Y, K157A, K157C, K157P, K157Q, K157S, K157V, G160A, G160L, G160Q, G160S, G160V, G160Y, A163D, A163G, A163K, A163N, A163Q, A163W, Q164D / L173M, Q164D / A188G, Q164D / Q191 R, Q164D / S192E, Q164D / Q195R, Q164D / S199G, Q164D / S199L, Q164D / V205L / K249E, Q164D / K209R, A165G, A165T, S166M, S166V, H167L, H167R, H167V, C169I, C169S, C169T, Q1711, Q171 R, Q171 S, Q171 V, R172I, R172T, R172V, L173I, M177G, M177R, M177S, M177T, M177V, M177W, M177Y, S182L, E187G, A188G, I190M, Q191 F, Q191 G, S192A, S192L, L194W, Q195G, Q195H, Q195M, Q195W, Q195Y, K197L / A266V, Q198A, Q198L, Q198N, Q198R, S199F, S199G, S199L, S199W, S199Y, H200A, H200F, H200L, H200R, H200S, H200V, H200Y, E202L, R203Q, F204I, F204T, F204V, V205T, A208S, K209V, H210T, H210V, V324M, W331 L, and E349K.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 1580 and one or more residue differences as compared to SEQ ID NO: 1580, selected from 73, 91 , 93, 95, 97, 98, 110, 112, 122, 122 / 164, 124, 127, 139, 144, 145, 146, 146 / 164, 147, 148, 150, 150 / 164, 151 , 151 / 164, 153, 153 / 164, 155, 156, 157, 159, 160, 161 , 162, 163, 164, 164 / 182, 164 / 188, 164 / 191 , 164 / 192, 164 / 195, 164 / 199, 164 / 205 / 249, 164 / 209, 165, 166, 167, 169, 171 , 172, 177, 184, 187, 188, 190, 191 , 192, 193, 195, 197 / 266, 198, 199, 200, 204, 205, 206, 207, 208, 209, 210, and 324. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 1580 and one or more residue differences as compared to SEQ ID NO: 1580, selected from 73Q, 73W, 91 C, 91 L, 93S, 95G, 95R, 95S, 95V, 97T, 98V, 110T, 110V, 112A, 112H, 122C, 122F, 122L, 122V, 122V / 164D, 124A, 124L,124M, 1271, 1391, 144F, 144Y, 145F, 145L, 145Q, 145R, 145S, 145T, 146A, 146F, 146G, 146K, 146L / 164D, 146Q, 146R, 146V, 147W, 148E, 148F, 148G, 148R, 148V, 150A, 150G, 150G / 164D, 150L, 150R / 164D, 150S / 164D, 150T, 150V, 150W, 151 A, 151 C, 151 E, 151 G, 151 1, 151 K, 151 L, 151 L / 164D, 151 M, 151 N, 151 Q, 151 S, 151 T, 151 V, 151 W, 151 Y, 151 Y / 164D, 153G, 153G / 164D, 153P, 153Q, 153R, 155L, 155S, 155W, 156G, 156S, 156Y, 157A, 157C, 157H, 157P, 157R, 157S, 157V, 159A, 159G, 159P, 159Q, 159T, 160A, 160L, 160M, 160S, 160V, 160Y, 161 A, 161 C, 161 F, 161 L, 161 M, 161 T, 162Y, 163D, 163G, 163W, 164D, 164D / 182W, 164D / 188G, 164D / 191 R, 164D / 192E, 164D / 195R, 164D / 199G, 164D / 199L, 164D / 205L / 249E, 164D / 209R, 165G, 165T, 165V, 166M, 166V, 166W, 167L, 1691, 169T, 171 R, 171 S, 171 V, 1721, 177W, 184Q, 184T, 187G, 188G, 190D, 190G, 190M, 190W, 191 F, 191 L, 191 W, 192A, 193W, 195H, 195R, 195Y, 197L / 266V, 198A, 198L, 198N, 198R, 198V, 199G, 199L, 199W, 199Y, 200R, 204L, 205T, 206L, 207K, 207Q, 208S, 208V, 209A, 209G, 209V, 21 OP, 21 OR, 21 OT, 210V, and 324M. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 1580 and one or more residue differences as compared to SEQ ID NO: 1580, selected from K73Q, K73W, A91 C, A91 L, D93S, T95G, T95R, T95S, T95V, S97T, L98V, K1 10T, K1 10V, G1 12A, G1 12H, W122C, W122F, W122L, W122V, W122V / Q164D, H124A, H124L, H124M, L127I, L139I, W144F, W144Y, W145F, W145L, W145Q, W145R, W145S, W145T, M146A, M146F, M146G, M146K, M146L / Q164D, M146Q, M146R, M146V, R147W, P148E, P148F, P148G, P148R, P148V, H150A, H150G, H150G / Q164D, H150L, H150R / Q164D, H150S / Q164D, H150T, H150V, H150W, P151 A, P151 C, P151 E, P151 G, P151 1, P151 K, P151 L, P151 L / Q164D, P151 M, P151 N, P151 Q, P151 S, P151 T, P151 V, P151 W, P151 Y, P151 Y / Q164D, D153G, D153G / Q164D, D153P, D153Q, D153R, D155L, D155S, D155W, E156G, E156S, E156Y, K157A, K157C, K157H, K157P, K157R, K157S, K157V, S159A, S159G, S159P, S159Q, S159T, G160A, G160L, G160M, G160S, G160V, G160Y, K161 A, K161 C, K161 F, K161 L, K161 M, K161 T, W162Y, A163D, A163G, A163W, Q164D, Q164D / S182W, Q164D / A188G, Q164D / Q191 R, Q164D / S192E, Q164D / Q195R, Q164D / S199G, Q164D / S199L, Q164D / V205L / K249E, Q164D / K209R, A165G, A165T, A165V, S166M, S166V, S166W, H167L, C169I, C169T, Q171 R, Q171 S, Q171 V, R172I, M177W, C184Q, C184T, E187G, A188G, I190D, I190G, I190M, I190W, Q191 F, 0191 L, Q191 W, S192A, Y193W, Q195H, Q195R, Q195Y, K197L / A266V, Q198A, Q198L, Q198N, Q198R, Q198V, S199G, S199L, S199W, S199Y, H200R, F204L, V205T, V206L, R207K, R207Q, A208S, A208V, K209A, K209G, K209V, H210P, H210R, H210T, H210V, and V324M.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 1580 and one or more residue differences as compared to SEQ ID NO: 1580, selected from 69, 73, 91 , 127, 140, 144, 145, 146, 146 / 164, 150, 150 / 164, 151 , 153, 155, 156, 157, 160, 163, 164, 164 / 172, 164 / 188, 164 / 191 , 164 / 192, 164 / 195, 164 / 199, 164 / 209, 165, 167, 169, 171 , 172, 182, 183, 188, 191 , 192, 193, 195, 196, 198, 199, 200, 204, 205, 208, 210, and 324. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or moresequence identity to reference sequence SEQ ID NO: 1580 and one or more residue differences as compared to SEQ ID NO: 1580, selected from 69G, 69V, 73Q, 73W, 91 G, 1271, 140V, 144F, 144Y, 145R, 146A, 146F, 146K, 146L / 164D, 146Q, 146R, 150G, 150L, 150R, 150R / 164D, 150S / 164D, 150W, 151 K, 151 N, 153G, 153Q, 153R, 155C, 155F, 155L, 155M, 155P, 155S, 155T, 155W, 156C, 156G, 156R, 156S, 156T, 156W, 156Y, 157A, 157P, 157R, 157S, 157V, 160A, 160L, 160M, 160Q, 160R, 160S, 160V, 160Y, 163D, 163K, 163N, 163W, 164D, 164D / 172M, 164D / 188G, 164D / 191 R, 164D / 192E, 164D / 195R, 164D / 199G, 164D / 199L, 164D / 209R, 165G, 167L, 167R, 169A, 1691, 169T, 171 H, 171 R, 171 S, 171 V, 1721, 182T, 183L, 188G, 191 F, 191 G, 191 W, 192A, 192L, 192R, 193W, 195G, 195H, 195M, 195R, 195W, 195Y, 196C, 196G, 196L, 196W, 198A, 198L, 198N, 198R, 198V, 199L, 199W, 199Y, 200R, 200Y, 204L, 205T, 208S, 210V, and 324M. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 1580 and one or more residue differences as compared to SEQ ID NO: 1580, selected from E69G, E69V, K73Q, K73W, A91 G, L127I, 1140V, W144F, W144Y, W145R, M146A, M146F, M146K, M146L / Q164D, M146Q, M146R, H150G, H150L, H150R, H150R / Q164D, H150S / Q164D, H150W, P151 K, P151 N, D153G, D153Q, D153R, D155C, D155F, D155L, D155M, D155P, D155S, D155T, D155W, E156C, E156G, E156R, E156S, E156T, E156W, E156Y, K157A, K157P, K157R, K157S, K157V, G160A, G160L, G160M, G160Q, G160R, G160S, G160V, G160Y, A163D, A163K, A163N, A163W, Q164D, Q164D / R172M, Q164D / A188G, Q164D / Q191 R, Q164D / S192E, Q164D / Q195R, Q164D / S199G, Q164D / S199L, Q164D / K209R, A165G, H167L, H167R, C169A, C169I, C169T, Q171 H, Q171 R, Q171 S, Q171 V, R172I, S182T, V183L, A188G, Q191 F, Q191 G, Q191 W, S192A, S192L, S192R, Y193W, Q195G, Q195H, Q195M, Q195R, Q195W, Q195Y, D196C, D196G, D196L, D196W, Q198A, Q198L, Q198N, Q198R, Q198V, S199L, S199W, S199Y, H200R, H200Y, F204L, V205T, A208S, H210V, and V324M.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 972 and one or more residue differences as compared to SEQ ID NO: 972, selected from 38 / 143 / 168 / 220 / 290 / 317 / 423, 38 / 152 / 154 / 164 / 168 / 220 / 290 / 317 / 423, 38 / 152 / 168 / 220 / 290 / 317 / 343 / 423, 38 / 152 / 168 / 220 / 290 / 317 / 423, 38 / 154 / 164 / 168 / 220 / 290 / 317 / 423, 38 / 168 / 220 / 290 / 317 / 423, and 38 / 220 / 317. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 972 and one or more residue differences as compared to SEQ ID NO: 972, selected from 38F / 143R / 168Q / 220R / 290R / 317M / 423D, 38F / 152T / 154K / 164R / 168K / 220R / 290R / 317M / 423D, 38F / 152T / 168Q / 220R / 290R / 317M / 343R / 423D, 38F / 152T / 168Q / 220R / 290R / 317M / 423D, 38F / 154K / 164Q / 168Q / 220R / 290R / 317M / 423D, 38F / 168Q / 220R / 290R / 317M / 423D, and 38F / 220R / 317M. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 972 and one or more residue differences as compared to SEQ ID NO:972, selected from C38F / D143R / F168Q / P220R / K290R / R317M / W423D,C38F / E152T / A154K / D164R / F168K / P220R / K290R / R317M / W423D,C38F / E152T / F168Q / P220R / K290R / R317M / E343R / W423D, C38F / E152T / F168Q / P220R / K290R / R317M / W423D,C38F / A154K / D164Q / F168Q / P220R / K290R / R317M / W423D, C38F / F168Q / P220R / K290R / R317M / W423D, and C38F / P220R / R317M.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 2368 and one or more residue differences as compared to SEQ ID NO: 2368, selected from 73 / 143 / 169 / 191 / 193, 73 / 156, 73 / 156 / 192 / 435, 73 / 156 / 210, 73 / 163, 73 / 163 / 210, 73 / 192 / 193, 127, 127 / 160 / 172 / 198, 127 / 172, 127 / 198, 146, 146 / 151 / 188, 146 / 324, 151 / 171 / 324, 151 / 324, 156 / 169, 156 / 192, 157 / 171 , 163, 163 / 169, 163 / 169 / 191 / 192 / 193, 165 / 171 / 324, 165 / 188 / 324, 165 / 324, 169, 171 / 188 / 324, 171 / 324, 172, 188, 192, 192 / 193, 198, 210, and 324. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 2368 and one or more residue differences as compared to SEQ ID NO: 2368, selected from 73Q / 143K / 1691 / 191 F / 193W, 73Q / 156R, 73Q / 156R / 210V, 73Q / 156Y / 192A / 435K, 73Q / 163D, 73Q / 163D / 210Q, 73Q / 163N, 73Q / 163N / 210V, 73Q / 192A / 193W, 1271, 1271 / 160A / 1721 / 198R, 1271 / 1721, 1271 / 198R, 146L, 146L / 151 N / 188G , 146L / 324M, 151 N / 171 V / 324M, 151 N / 324M, 156Y / 169I, 156Y / 192E, 157P / 171 V, 163D, 163D / 1691 / 191 F / 192A / 193W, 163N / 169I, 165G / 171 R / 324M, 165G / 188G / 324M, 165G / 324M, 1691, 171 R / 324M, 171 V / 188G / 324M, 171 V / 324M, 1721, 188G, 192A, 192E / 193W, 198R, 210V, and 324M. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 2368 and one or more residue differences as compared to SEQ ID NO: 2368, selected from K73Q / D143K / C169I / Q191 F / Y193W, K73Q / E156R, K73Q / E156R / H210V, K73Q / E156Y / S192A / M435K, K73Q / A163D, K73Q / A163D / H210Q, K73Q / A163N, K73Q / A163N / H210V, K73Q / S192A / Y193W, L127I, L127I / G160A / R172I / Q198R, L127I / R172I, L127I / Q198R, M146L, M146L / P151 N / A188G, M146L / V324M, P151 N / Q171 V / V324M, P151 N / V324M, E156Y / C169I, E156Y / S192E, K157P / Q171 V, A163D, A163D / C169I / Q191 F / S192A / Y193W, A163N / C169I, A165G / Q171 R / V324M, A165G / A188G / V324M, A165G / V324M, C169I, Q171 R / V324M, Q171 V / A188G / V324M, Q171 V / V324M, R172I, A188G, S192A, S192E / Y193W, Q198R, H210V, and V324M.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 2368 and one or more residue differences as compared to SEQ ID NO: 2368, selected from 73, 73 / 163 / 191 / 192 / 193, 127 / 160 / 172, 127 / 160 / 172 / 198, 144, 160, 160 / 172, 163 / 191 / 192, and 192. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%,96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 2368 and one or more residue differences as compared to SEQ ID NO: 2368, selected from 73Q,73Q / 163N / 191 F / 192A / 193W, 1271 / 160A / 172I, 1271 / 160A / 1721 / 198R, 144Y, 160A, 160A / 172I, 163N / 191 F / 192A, and 192A. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 2368 and one or more residue differences as compared to SEQ ID NO: 2368, selected from K73Q, K73Q / A163N / Q191 F / S192A / Y193W, L127I / G160A / R172I, L127I / G160A / R172I / Q198R, W144Y, G160A, G160A / R172I, A163N / Q191 F / S192A, and S192A.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 2426 and one or more residue differences as compared to SEQ ID NO: 2426, selected from 8, 13, 17, 18, 20 / 146 / 157 / 167, 26, 34 / 38, 38 / 39 / 45, 67, 70, 73, 73 / 150 / 155 / 169, 73 / 150 / 171 / 192, 73 / 155, 73 / 155 / 169 / 199, 73 / 155 / 171 , 73 / 192, 73 / 199, 78, 82, 84, 86, 86 / 384, 89, 90, 91 , 95, 102, 110, 111 , 113, 114, 115, 122 / 127, 126 / 127, 127 / 131 / 165, 127 / 133, 143, 143 / 146, 143 / 146 / 153, 143 / 146 / 157, 143 / 146 / 167, 143 / 146 / 167 / 188, 143 / 146 / 188, 143 / 153 / 167 / 188 / 246, 143 / 156 / 157 / 167 / 195, 143 / 156 / 188, 143 / 188, 143 / 188 / 195, 143 / 195, 144, 145, 145 / 151 , 146, 146 / 152, 146 / 153 / 157, 146 / 153 / 157 / 167, 146 / 153 / 167, 146 / 157, 146 / 157 / 167, 146 / 167, 150, 150 / 155 / 169, 150 / 155 / 169 / 192, 150 / 155 / 192, 150 / 169, 150 / 192, 150 / 199, 151 / 152 / 154, 152 / 154, 152 / 154 / 155 / 160, 153 / 157 / 167, 155 / 171 , 155 / 192, 156 / 270, 160 / 371 , 167 / 188, 168 / 172, 169, 184, 190, 192 / 198, 198, 206, 208, 209, 210, 214 / 220, 215 / 220, 216 / 220, 217 / 220, 219 / 220, 220, 220 / 222, 220 / 225, 220 / 226, 231 , 235, 242, 257, 259, 261 , 262, 267, 268, 269, 270, 272, 274, 275, 276, 277, 278, 282, 283 / 290, 284 / 290, 287 / 290, 288 / 290, 289 / 290, 290, 290 / 291 , 290 / 292, 290 / 293, 290 / 294, 304, 305, 305 / 373, 306, 307, 309, 317, 329, 333, 334, 339, 341 , 344, 345, 346, 352, 353, 355, 358, 366, 368, 370, 372, 373, 375, 376, 378, 379, 380, 381 , 382, 383, 384, 385, 386 / 393, 387 / 393, 389 / 393, 390 / 393, 391 / 393, 392 / 393, 393 / 394, 414, 415, 420 / 423, 422 / 423, 423, 423 / 425, 432, 440, 441 , 443, 469, and 472. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 2426 and one or more residue differences as compared to SEQ ID NO: 2426, selected from 8D, 8V, 13W, 17V, 18D, 18L, 18P, 20D / 146C / 157P / 167L, 26R, 34H / 38C, 38C / 39F / 45I, 38C / 39P / 45I, 67G, 70G, 73H, 73Q / 150N / 155L / 169T, 73Q / 150N / 171 R / 192A, 73Q / 155L, 73Q / 155L / 169T / 199L, 73Q / 155L / 171 R, 73Q / 192A, 73Q / 199L, 73R, 78W, 82Q, 82R, 84T, 86F / 384T, 86I, 89M, 90W, 91 L, 91 V, 95V, 102G, 110L, 111 M, 113A, 113G, 113V, 114D, 114T, 115Q, 122A / 127L, 126V / 127L, 127L / 131 A / 165T, 127L / 133M, 143T, 143V, 143V / 146C, 143V / 146C / 153G, 143V / 146C / 157P, 143V / 146C / 167L, 143V / 146C / 167L / 188G, 143V / 146C / 188G, 143V / 146K, 143V / 153G / 167L / 188G / 246R, 143V / 156R / 157P / 167L / 195H, 143V / 156R / 188G, 143V / 188G, 143V / 188G / 195H, 143V / 195H, 144F, 145L, 145V / 151 R, 146A / 152A, 146C, 146C / 153G / 157P, 146C / 153G / 157P / 167L, 146C / 153G / 167L, 146C / 157P, 146C / 157P / 167L, 146C / 167L, 146K, 146K / 157P, 150N, 150N / 155L / 169T, 150N / 155L / 169T / 192A, 150N / 155L / 192A, 150N / 169T, 150N / 192A, 150N / 199L, 151 E / 152E / 154A, 151 R / 152E / 154A,151T / 152E / 154A, 152E / 154A, 152E / 154A / 155N / 160G, 153G / 157P / 167L, 155L / 171 R, 155L / 192A, 156G / 270P, 160G / 371 W, 167L / 188G, 168F / 172V, 169T, 184A, 190G, 190S, 192L / 198Q, 198D, 198G, 206F, 206W, 208V, 209G, 209M, 210G, 21 OL, 214G / 220P, 214V / 220P, 215F / 220P, 216I / 220P, 217D / 220P, 217G / 220P, 217L / 220P, 217R / 220P, 219G / 220P, 220L, 220P / 222C, 220P / 222H, 220P / 222R, 220P / 225M, 220P / 226R, 220P / 226S, 231 G, 235S, 242S, 257G, 259H, 261 K, 262L, 267L, 268W, 269A, 269E, 269G, 270D, 270L, 270P, 272G, 272R, 272T, 274K, 274R, 275R, 275W, 276A, 276E, 276G, 277V, 278V, 282M, 283S / 290K, 284A / 290K, 284S / 290K, 287P / 290K, 288M / 290K, 288P / 290K, 288R / 290K, 288V / 290K, 289S / 290K, 290C, 290K, 290K / 291 E, 290K / 291G, 290K / 291 S, 290K / 292A, 290K / 293L, 290K / 294K, 290K / 294M, 290K / 294R, 290K / 294S, 290K / 294W, 290Q, 290V, 304E, 304V, 305G / 373C, 305R, 306G, 307S, 309V, 317R, 329A, 329R, 329T, 333L, 334L, 339G, 341 L, 344G, 345T, 346T, 346V, 352L, 353V, 355C, 358C, 366S, 368W, 370S, 372G, 373T, 375Y, 376L, 378A, 378G, 378P, 378S, 379P, 380T, 380V, 381 S, 382L, 383F, 383R, 384N, 384V, 385L, 386A / 393E, 386S / 393E, 386T / 393E, 386V / 393E, 387A / 393E, 387R / 393E, 389L / 393E, 389M / 393E, 389V / 393E, 390E / 393E, 390G / 393E, 391 L / 393E, 392P / 393E, 393E / 394R, 414G, 415P, 415V, 420E / 423W, 422G / 423W, 422S / 423W, 422V / 423W, 423A, 423W / 425A, 432T, 440G, 441 Q, 441 S, 441 W, 443P, 469A, 472A, and 472R. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 2426 and one or more residue differences as compared to SEQ ID NO: 2426, selected from Q8D, Q8V, H13W, A17V, A18D, A18L, A18P, G20D / M146C / K157P / H167L, K26R, D34H / F38C, F38C / S39F / V45I, F38C / S39P / V45I, S67G, V70G, K73H, K73Q / H150N / D155L / C169T, K73Q / H150N / Q171 R / S192A, K73Q / D155L, K73Q / D155L / C169T / S199L, K73Q / D155L / Q171 R, K73Q / S192A, K73Q / S199L, K73R, H78W, L82Q, L82R, Q84T, Y86F / A384T, Y86I, I89M, L90W, A91 L, A91 V, T95V, H102G, K110L, L111 M, S113A, S113G, S113V, V114D, V114T, K115Q, W122A / I127L, T126V / I127L, I127L / G131 A / A165T, I127L / T133M, D143T, D143V, D143V / M146C, D143V / M146C / D153G, D143V / M146C / K157P, D143V / M146C / H167L, D143V / M146C / H167L / A188G, D143V / M146C / A188G, D143V / M146K, D143V / D153G / H167L / A188G / S246R, D143V / E156R / K157P / H167UQ195H, D143V / E156R / A188G, D143V / A188G, D143V / A188G / Q195H, D143V / Q195H, W144F, W145L, W145V / P151 R, M146A / T152A, M146C, M146C / D153G / K157P, M146C / D153G / K157P / H167L, M146C / D153G / H167L, M146C / K157P, M146C / K157P / H167L, M146C / H167L, M146K, M146K / K157P, H150N, H150N / D155L / C169T, H150N / D155L / C169T / S192A, H150N / D155L / S192A, H150N / C169T, H150N / S192A, H150N / S199L, P151 E / T152E / K154A, P151 R / T152E / K154A, P151 T / T152E / K154A, T152E / K154A, T152E / K154A / D155N / A160G, D153G / K157P / H167L, D155L / Q171 R, D155L / S192A, E156G / K270P, A160G / P371W, H167L / A188G, K168F / I172V, C169T, C184A, I190G, 1190S, S192L / R198Q, R198D, R198G, V206F, V206W, A208V, K209G, K209M, H210G, H210L, I214G / R220P, I214V / R220P, L215F / R220P, D216I / R220P, S217D / R220P, S217G / R220P, S217L / R220P, S217R / R220P, L219G / R220P, R220L, R220P / F222C, R220P / F222H, R220P / F222R, R220P / L225M, R220P / E226R, R220P / E226S, L231 G, T235S, G242S, K257G, S259H, S261 K, V262L, V267L, T268W, L269A, L269E, L269G, K270D, K270L, K270P, K272G, K272R, K272T, E274K, E274R, M275R, M275W, Q276A, Q276E, Q276G, P277V, I278V, Y282M, A283S / R290K, E284A / R290K, E284S / R290K, N287P / R290K,S288M / R290K, S288P / R290K, S288R / R290K, S288V / R290K, P289S / R290K, R290C, R290K, R290K / D291 E, R290K / D291 G, R290K / D291 S, R290K / M292A, R290K / E293L, R290K / P294K, R290K / P294M, R290K / P294R, R290K / P294S, R290K / P294W, R290Q, R290V, P304E, P304V, I305G / Y373C, I305R, E306G, T307S, S309V, M317R, K329A, K329R, K329T, T333L, G334L, R339G, R341 L, E344G, P345T, R346T, R346V, V352L, S353V, L355C, I358C, R366S, A368W, T370S, P372G, Y373T, L375Y, R376L, K378A, K378G, K378P, K378S, G379P, L380T, L380V, V381 S, D382L, A383F, A383R, A384N, A384V, N385L, E386A / A393E, E386S / A393E, E386T / A393E, E386V / A393E, L387A / A393E, L387R / A393E, S389L / A393E, S389M / A393E, S389V / A393E, Q390E / A393E, Q390G / A393E, S391 L / A393E, C392P / A393E, A393E / T394R, N414G, G415P, G415V, S420E / D423W, K422G / D423W, K422S / D423W, K422V / D423W, D423A, D423W / Y425A, G432T, T440G, G441 Q, G441 S, G441 W, A443P, L469A, G472A, and G472R.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 2426 and one or more residue differences as compared to SEQ ID NO: 2426, selected from 8, 18, 25, 29, 30, 67, 73, 73 / 150 / 155 / 169, 73 / 150 / 155 / 171 , 73 / 150 / 171 / 192, 73 / 171 , 73 / 192, 73 / 199, 74, 82, 86, 86 / 384, 113, 115, 118, 127 / 128, 143, 150, 150 / 155 / 169, 150 / 155 / 169 / 192, 150 / 155 / 171 , 150 / 155 / 171 / 199, 150 / 155 / 192, 150 / 155 / 199, 150 / 192, 150 / 199, 155 / 192, 168 / 171 / 172, 195 / 198, 197 / 198, 198 / 201 , 198 / 203, 198 / 205, 235, 252, 256, 257, 262, 263, 267 / 343, 268, 269, 270, 272, 273, 276, 277, 288 / 290, 289 / 290, 290, 290 / 291 , 290 / 292, 290 / 294, 298, 304, 305, 307, 333, 346, 355, 366, 370, 386 / 393, 389 / 393, 393 / 395, 440, and 472. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 2426 and one or more residue differences as compared to SEQ ID NO: 2426, selected from 8D, 18L, 25G, 29F, 30R, 67G, 73H, 73Q / 150N / 155L / 169T, 73Q / 150N / 155L / 171 R, 73Q / 150N / 171 R / 192A, 73Q / 171 R, 73Q / 192A, 73Q / 199L, 74R, 82F, 82Q, 86A, 86F / 384T, 86I, 113E, 115A, 115Q, 118S, 127L / 128I, 143H, 150N, 150N / 155L / 169T, 150N / 155L / 169T / 192A, 150N / 155L / 171 R, 150N / 155L / 171 R / 199L, 150N / 155L / 192A, 150N / 155L / 199L, 150N / 192A, 150N / 199L, 155L / 192A, 168F / 171 P / 172R, 195K / 198Q, 195S / 198Q, 197S / 198Q, 198Q / 201 R, 198Q / 203K, 198Q / 205T, 235S, 252A, 256E, 256G, 257G, 262L, 263Q, 267P / 343G, 268W, 269I, 270P, 272R, 273V, 276E, 277V, 288V / 290K, 289S / 290K, 290K / 291 E, 290K / 292A, 290K / 294S, 290Q, 290V, 298R, 304W, 305A, 305R, 307S, 333L, 346T, 355K, 366S, 370S, 386S / 393E, 389M / 393E, 393E / 395R, 393E / 395W, 440G, and 472R. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 2426 and one or more residue differences as compared to SEQ ID NO: 2426, selected from Q8D, A18L, T25G, I29F, E30R, S67G, K73H, K73Q / H150N / D155L / C169T, K73Q / H150N / D155L / Q171 R, K73Q / H150N / Q171 R / S192A, K73Q / Q171 R, K73Q / S192A, K73Q / S199L, A74R, L82F, L82Q, Y86A, Y86F / A384T, Y86I, S113E, K115A, K115Q, A118S, I127L / L128I, D143H, H150N, H150N / D155L / C169T, H150N / D155L / C169T / S192A, H150N / D155L / Q171 R, H150N / D155L / Q171 R / S199L,H150N / D155L / S192A, H150N / D155L / S199L, H150N / S192A, H150N / S199L, D155L / S192A,K168F / Q171 P / 1172R, Q195K / R198Q, Q195S / R198Q, K197S / R198Q, R198Q / N201 R, R198Q / R203K, R198Q / V205T, T235S, N252A, R256E, R256G, K257G, V262L, K263Q, V267P / E343G, T268W, L269I, K270P, K272R, D273V, Q276E, P277V, S288V / R290K, P289S / R290K, R290K / D291 E, R290K / M292A, R290K / P294S, R290Q, R290V, V298R, P304W, I305A, I305R, T307S, T333L, R346T, L355K, R366S, T370S, E386S / A393E, S389M / A393E, A393E / V395R, A393E / V395W, T440G, and G472R.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 2540 and one or more residue differences as compared to SEQ ID NO: 2540, selected from 66, 67, 70, 101 , 102, 103, 104, 106, 107, 108, 113, 114, 115, 117, 118, 143, 152, 154, 235, 236 / 251 , 237, 238, 242, 243, 246, 247, 249, 250, 251 , 256 / 352, 257, 259, 285, 286, 287, 288, 289, 290, 292, 293, 294, 295, 296, 322, 323, 326, 329, 330, 333, 339, 340, 343, 344, 345, and 352. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 2540 and one or more residue differences as compared to SEQ ID NO: 2540, selected from 66T, 67V, 70A, 101 S, 101 T, 102R, 102S, 103V, 104T, 106A, 107P, 107S, 107V, 108M, 113G, 113R, 114A, 1141, 114P, 114S, 115L, 115R, 117A, 118K, 143E, 143G, 143R, 143V, 152R, 154E, 154F, 154Q, 235R, 236V / 251 T, 237S, 238L, 242T, 243F, 243L, 246M, 247F, 249F, 249L, 249R, 249V, 249W, 250N, 251 G, 251 R, 251 S, 256K / 352A, 257L, 257R, 257V, 259H, 259P, 259Y, 285D, 286G, 286R, 287A, 287G, 287L, 287R, 287V, 288L, 288W, 289I, 289K, 289R, 289S, 290G, 290S, 292L, 292P, 292V, 293G, 294E, 294V, 295F, 295T, 295V, 296A, 296Q, 296R, 322G, 322Q, 322T, 323S, 323T, 326L, 326M, 329G, 329H, 330L, 330V, 333A, 333M, 333R, 333S, 339A, 339G, 339S, 340S, 343P, 343T, 344G, 344M, 345A, 345E, 345K, 345V, 352I, and 352L. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 2540 and one or more residue differences as compared to SEQ ID NO: 2540, selected from V66T, S67V, V70A, K101 S, K101T, H102R, H102S, Q103V, V104T, S106A, D107P, D107S, D107V, L108M, S113G, S113R, V114A, V114I, V114P, V114S, K115L, K115R, K117A, A118K, D143E, D143G, D143R, D143V, T152R, K154E, K154F, K154Q, T235R, I236V / K251T, D237S, I238L, G242T, V243F, V243L, S246M, K247F, K249F, K249L, K249R, K249V, K249W, R250N, K251 G, K251 R, K251 S, R256K / V352A, K257L, K257R, K257V, S259H, S259P, S259Y, E285D, T286G, T286R, N287A, N287G, N287L, N287R, N287V, S288L, S288W, P289I, P289K, P289R, P289S, R290G, R290S, M292L, M292P, M292V, E293G, P294E, P294V, L295F, L295T, L295V, K296A, K296Q, K296R, W322G, W322Q, W322T, R323S, R323T, E326L, E326M, K329G, K329H, A330L, A330V, T333A, T333M, T333R, T333S, R339A, R339G, R339S, L340S, E343P, E343T, E344G, E344M, P345A, P345E, P345K, P345V, V352I, and V352L.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to referencesequence SEQ ID NO: 2540 and one or more residue differences as compared to SEQ ID NO: 2540, selected from 18, 18 / 113 / 270, 18 / 113 / 270 / 329 / 370, 18 / 113 / 270 / 370 / 384, 18 / 113 / 270 / 370 / 412, 18 / 113 / 329 / 384, 18 / 259 / 370 / 384, 18 / 270, 18 / 270 / 370, 18 / 270 / 370 / 384, 18 / 370, 18 / 370 / 384, 73, 73 / 115, 73 / 115 / 184 / 333, 73 / 115 / 184 / 341 , 73 / 115 / 256, 73 / 184 / 256, 73 / 184 / 273 / 333, 73 / 184 / 333, 73 / 256, 73 / 256 / 375, 73 / 273, 73 / 273 / 333 / 375, 73 / 273 / 375, 73 / 333, 73 / 333 / 375, 73 / 375, 73 / 379, 77 / 233 / 238 / 376, 77 / 233 / 334 / 376, 77 / 262 / 376, 77 / 276, 77 / 334 / 376, 82, 82 / 127, 82 / 127 / 144 / 235 / 290, 82 / 127 / 144 / 266, 82 / 127 / 144 / 266 / 290, 82 / 127 / 144 / 378, 82 / 127 / 235, 82 / 127 / 290, 82 / 144 / 266 / 290 / 378, 82 / 144 / 266 / 378, 82 / 235 / 266, 82 / 235 / 266 / 378 / 383, 82 / 235 / 290, 82 / 235 / 378, 82 / 266, 82 / 266 / 378, 82 / 378, 113 / 270 / 370 / 384, 115 / 256 / 333 / 375, 115 / 256 / 375, 115 / 273 / 375, 115 / 375, 127 / 144 / 290, 127 / 149 / 150, 127 / 266 / 290 / 378, 127 / 378 / 383, 144 / 235 / 290, 144 / 266 / 290, 233 / 334, 233 / 376, 235, 235 / 290, 256 / 333, 262, 266 / 378, 270 / 370, 273, 276 / 352, 290, 290 / 378, 333, 358, 370, 375, and 378. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 2540 and one or more residue differences as compared to SEQ ID NO: 2540, selected from 18D, 18D / 113A / 270L, 18D / 113A / 270L / 329A / 370S, 18D / 259R / 370S / 384V, 18D / 270L, 18D / 270L / 370S, 18D / 270U370S / 384V, 18D / 370S, 18L, 18L / 113A / 270L, 18L / 113A / 270L / 370S / 384V, 18L / 113A / 329A / 384V, 18L / 113E / 270L / 370S / 412S, 18L / 370S, 18L / 370S / 384V, 73H, 73H / 115Q / 184A / 341 G, 73H / 184A / 256G, 73H / 273V / 333L / 375S, 73H / 333L, 73R, 73R / 115Q, 73R / 115Q / 184A / 333L, 73R / 115Q / 256G, 73R / 184A / 273V / 333L, 73R / 184A / 333L, 73R / 256G, 73R / 256G / 375S, 73R / 273V, 73R / 273V / 333L / 375S, 73R / 273V / 375S, 73R / 333L, 73R / 333L / 375S, 73R / 375S, 73R / 379V, 77V / 233U238V / 376L, 77V / 233L / 334L / 376L, 77V / 262L / 376L, 77V / 276E, 77V / 334L / 376L, 82Q, 82Q / 127L, 82Q / 127L / 144Y / 235S / 290V, 82Q / 127L / 144Y / 266V, 82Q / 127L / 144 Y / 266 V / 290 V, 82Q / 127L / 144Y / 378S, 82Q / 127L / 235S, 82Q / 127L / 290Q, 82Q / 144Y / 266V / 290Q / 378S, 82Q / 144Y / 266V / 378S, 82Q / 235S / 266V, 82Q / 235S / 266V / 378S / 383P, 82Q / 235S / 290V, 82Q / 235S / 378S, 82Q / 266V, 82Q / 266V / 378S, 82Q / 378S, 113A / 270L / 370S / 384V, 115Q / 256G / 333L / 375S, 115Q / 256G / 375S, 115Q / 273V / 375S, 115Q / 375S, 127L / 144Y / 290V, 127L / 149H / 150Q, 127L / 266V / 290Q / 378S, 127L / 378S / 383P, 144Y / 235S / 290Q, 144Y / 266V / 290Q, 233L / 334L, 233L / 376L, 235S, 235S / 290Q, 256G / 333L, 262L, 266V / 378S, 270L / 370S, 273V, 276E / 352L, 290Q, 290V, 290V / 378S, 333L, 358W, 370S, 375S, and 378S. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 2540 and one or more residue differences as compared to SEQ ID NO: 2540, selected from A18D, A18D / S113A / K270L,A18D / S113A / K270L / K329A / T370S, A18D / S259R / T370S / A384V, A18D / K270L, A18D / K270L / T370S, A18D / K270L / T370S / A384V, A18D / T370S, A18L, A18L / S113A / K270L, A18L / S113A / K270L / T370S / A384V, A18L / S113A / K329A / A384V, A18L / S113E / K270L / T370S / G412S, A18L / T370S, A18L / T370S / A384V, Q73H, Q73H / K115Q / C184A / R341 G, Q73H / C184A / R256G, Q73H / D273V / T333L / L375S, Q73H / T333L, Q73R, Q73R / K115Q, Q73R / K115Q / C184A / T333L, Q73R / K115Q / R256G, Q73R / C184A / D273V / T333L, Q73R / C184A / T333L, Q73R / R256G, Q73R / R256G / L375S, Q73R / D273V, Q73R / D273V / T333L / L375S, Q73R / D273V / L375S, Q73R / T333L, Q73R / T333L / L375S, Q73R / L375S, Q73R / G379V,E77V / A233L / I238V / R376L, E77V / A233L / G334L / R376L, E77V / V262L / R376L, E77V / Q276E, E77V / G334L / R376L, L82Q, L82Q / I127L, L82Q / I127L / W144Y / T235S / R290V, L82Q / I127L / W144Y / A266V, L82Q / I127L / W144Y / A266V / R290V, L82Q / I127L / W144Y / K378S, L82Q / I127L / T235S, L82Q / I127L / R290Q, L82Q / W144Y / A266V / R290Q / K378S, L82Q / W144Y / A266V / K378S, L82Q / T235S / A266V, L82Q / T235S / A266V / K378S / A383P, L82Q / T235S / R290V, L82Q / T235S / K378S, L82Q / A266V, L82Q / A266V / K378S, L82Q / K378S, S113A / K270L / T370S / A384V, K115Q / R256G / T333L / L375S,K115Q / R256G / L375S, K115Q / D273V / L375S, K115Q / L375S, I127L / W144Y / R290V, I127L / D149H / H150Q, I127L / A266V / R290Q / K378S, 1127L / K378S / A383P, W144Y / T235S / R290Q, W144Y / A266V / R290Q, A233L / G334L, A233L / R376L, T235S, T235S / R290Q, R256G / T333L, V262L, A266V / K378S, K270L / T370S, D273V, Q276E / V352L, R290Q, R290V, R290V / K378S, T333L, I358W, T370S, L375S, and K378S.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 2540 and one or more residue differences as compared to SEQ ID NO: 2540, selected from 18, 18 / 113, 18 / 113 / 252, 18 / 113 / 252 / 336 / 384, 18 / 113 / 252 / 370, 18 / 113 / 270, 18 / 113 / 270 / 370 / 384, 18 / 259 / 370 / 384, 18 / 270, 18 / 270 / 370, 18 / 270 / 370 / 384, 18 / 370, 18 / 370 / 384, 73, 73 / 115 / 184 / 341 , 73 / 115 / 341 , 73 / 131 / 256 / 273 / 333 / 341 / 375, 73 / 184 / 256, 73 / 184 / 256 / 341 , 73 / 184 / 256 / 375, 73 / 184 / 273 / 333, 73 / 184 / 333, 73 / 184 / 341 / 375, 73 / 256, 73 / 256 / 273, 73 / 256 / 273 / 375, 73 / 256 / 333 / 375, 73 / 256 / 375, 73 / 273, 73 / 273 / 333 / 341 , 73 / 273 / 333 / 375, 73 / 273 / 375, 73 / 333, 73 / 333 / 341 / 375, 73 / 333 / 375, 73 / 341 , 73 / 375, 73 / 379, 77, 77 / 118, 77 / 118 / 143 / 262 / 276 / 334 / 376, 77 / 118 / 143 / 334 / 352 / 376, 77 / 118 / 143 / 376, 77 / 118 / 233, 77 / 118 / 233 / 352 / 376, 77 / 118 / 262 / 334 / 376, 77 / 118 / 334, 77 / 143 / 262 / 276, 77 / 143 / 262 / 334, 77 / 143 / 376, 77 / 233 / 238 / 376, 77 / 233 / 262, 77 / 233 / 276 / 376, 77 / 233 / 334 / 376, 77 / 262 / 376, 77 / 276, 77 / 334 / 376, 77 / 352, 82, 82 / 127, 82 / 127 / 144 / 235 / 290, 82 / 127 / 144 / 266, 82 / 127 / 144 / 266 / 290, 82 / 127 / 144 / 378, 82 / 127 / 235, 82 / 127 / 235 / 266 / 358, 82 / 127 / 290, 82 / 127 / 290 / 358, 82 / 127 / 290 / 378, 82 / 144 / 266 / 290 / 378, 82 / 144 / 266 / 378, 82 / 150 / 235 / 266, 82 / 235 / 266, 82 / 235 / 266 / 290, 82 / 235 / 266 / 378 / 383, 82 / 235 / 290, 82 / 235 / 378, 82 / 266, 82 / 266 / 378, 82 / 290, 82 / 378, 113, 113 / 252, 113 / 252 / 270 / 370, 113 / 270 / 370 / 384, 113 / 336, 118 / 233 / 276 / 334, 118 / 334, 127 / 235 / 266, 127 / 266 / 290 / 378, 127 / 378 / 383, 143, 143 / 276, 143 / 276 / 334, 144 / 235 / 290, 144 / 266 / 290, 144 / 290 / 358, 233 / 334, 233 / 376, 235, 235 / 266 / 290 / 358 / 383, 235 / 358, 252 / 329 / 336 / 370, 252 / 384, 256 / 333, 256 / 375, 262, 262 / 276, 262 / 334, 266 / 290, 266 / 378, 270 / 370, 273, 276 / 352, 290, 290 / 378, 333, 334, 334 / 376, 336 / 370, 341 , 358, 370, 375, and 378. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 2540 and one or more residue differences as compared to SEQ ID NO: 2540, selected from 18D, 18D / 113A / 252A, 18D / 113A / 252A / 336Q / 384V, 18D / 113A / 270L, 18D / 259R / 370S / 384V, 18D / 270L, 18D / 270L / 370S, 18D / 270L / 370S / 384V, 18D / 370S, 18L, 18L / 113A, 18L / 113A / 270L, 18L / 113A / 270L / 370S / 384V, 18L / 113E, 18L / 113E / 252A / 370S, 18L / 370S, 18L / 370S / 384V, 73H, 73H / 115Q / 184A / 341 G, 73H / 115Q / 341 G, 73H / 184A / 256G, 73H / 256G / 273V / 375S, 73H / 256G / 333L / 375S, 73H / 273V / 333L / 341 G, 73H / 273V / 333L / 375S, 73H / 333L, 73R, 73R / 115Q / 341 G, 73R / 131 D / 256G / 273V / 333L / 341 G / 375S,73R / 184A / 256G / 341 G, 73R / 184A / 256G / 375S, 73R / 184A / 273V / 333L, 73R / 184A / 333L,73R / 184A / 341 G / 375S, 73R / 256G, 73R / 256G / 273V, 73R / 256G / 273V / 375S, 73R / 256G / 375S, 73R / 273V, 73R / 273V / 333L / 375S, 73R / 273V / 375S, 73R / 333L, 73R / 333L / 341 G / 375S, 73R / 333L / 375S, 73R / 341 G, 73R / 375S, 73R / 379V, 77V, 77V / 118S, 77V / 118S / 143H / 262L / 276E / 334L / 376L,77V / 118S / 143H / 334L / 352L / 376L, 77V / 118S / 143H / 376L, 77V / 118S / 233L, 77V / 118S / 233L / 352L / 376L, 77V / 118S / 262L / 334L / 376L, 77V / 118S / 334L, 77V / 143H / 262L / 276E, 77V / 143H / 262L / 334L, 77V / 143H / 376L, 77V / 233L / 238V / 376L, 77V / 233L / 262L, 77V / 233L / 276E / 376L, 77V / 233L / 334L / 376L, 77V / 262L / 376L, 77V / 276E, 77V / 334L / 376L, 77V / 352L, 82Q, 82Q / 127L, 82Q / 127L / 144Y / 235S / 290V, 82Q / 127L / 144Y / 266V, 82Q / 127L / 144 Y / 266 V / 290 V, 82Q / 127L / 144Y / 378S, 82Q / 127L / 235S, 82Q / 127L / 235S / 266V / 358W, 82Q / 127L / 290Q, 82Q / 127L / 290Q / 358W, 82Q / 127L / 290V / 378S, 82Q / 144Y / 266V / 290Q / 378S, 82Q / 144 Y / 266 V / 378S, 82Q / 150Q / 235S / 266V, 82Q / 235S / 266V, 82Q / 235S / 266V / 290Q, 82Q / 235S / 266V / 378S / 383P, 82Q / 235S / 290V, 82Q / 235S / 378S, 82Q / 266V, 82Q / 266V / 378S, 82Q / 290V, 82Q / 378S, 113A / 270L / 370S / 384V, 113E, 113E / 252A,113E / 252A / 270L / 370S, 113E / 336Q, 118S / 233L / 276E / 334L, 118S / 334L, 127L / 235S / 266V, 127L / 266V / 290Q / 378S, 127L / 378S / 383P, 143H, 143H / 276E, 143H / 276E / 334L, 144Y / 235S / 290Q, 144Y / 266V / 290Q, 144Y / 290V / 358W, 233L / 334L, 233L / 376L, 235S, 235S / 266V / 290Q / 358W / 383P, 235S / 358W, 252A / 329A / 336Q / 370S, 252A / 384V, 256G / 333L, 256G / 375S, 262L, 262L / 276E, 262L / 334L, 266V / 290Q, 266V / 378S, 270L / 370S, 273V, 276E / 352L, 290Q, 290V / 378S, 333L, 334L, 334L / 376L, 336Q / 370S, 341 G, 358W, 370S, 375S, and 378S. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 2540 and one or more residue differences as compared to SEQ ID NO: 2540, selected from A18D, A18D / S113A / N252A, A18D / S113A / N252A / G336Q / A384V, A18D / S113A / K270L, A18D / S259R / T370S / A384V, A18D / K270L, A18D / K270L / T370S, A18D / K270L / T370S / A384V,A18D / T370S, A18L, A18L / S113A, A18L / S113A / K270L, A18L / S113A / K270L / T370S / A384V, A18L / S113E, A18L / S113E / N252A / T370S, A18L / T370S, A18L / T370S / A384V, Q73H, Q73H / K115Q / C184A / R341 G, Q73H / K115Q / R341 G, Q73H / C184A / R256G, Q73H / R256G / D273V / L375S, Q73H / R256G / T333L / L375S, Q73H / D273V / T333L / R341 G, Q73H / D273V / T333L / L375S, Q73H / T333L, Q73R, Q73R / K115Q / R341 G, Q73R / G131 D / R256G / D273V / T333L / R341 G / L375S, Q73R / C184A / R256G / R341 G, Q73R / C184A / R256G / L375S, Q73R / C184A / D273V / T333L, Q73R / C184A / T333L,Q73R / C184A / R341 G / L375S, Q73R / R256G, Q73R / R256G / D273V, Q73R / R256G / D273V / L375S, Q73R / R256G / L375S, Q73R / D273V, Q73R / D273V / T333L / L375S, Q73R / D273V / L375S, Q73R / T333L, Q73R / T333L / R341 G / L375S, Q73R / T333L / L375S, Q73R / R341 G, Q73R / L375S, Q73R / G379V, E77V, E77V / A118S, E77V / A118S / D143H / V262L / Q276E / G334L / R376L,E77V / A118S / D143H / G334L / V352L / R376L, E77V / A118S / D143H / R376L, E77V / A118S / A233L, E77V / A118S / A233L / V352L / R376L, E77V / A118S / V262L / G334L / R376L, E77V / A118S / G334L, E77V / D143H / V262L / Q276E, E77V / D143H / V262L / G334L, E77V / D143H / R376L, E77V / A233L / I238V / R376L, E77V / A233L / V262L, E77V / A233L / Q276E / R376L, E77V / A233L / G334L / R376L, E77V / V262L / R376L, E77V / Q276E, E77V / G334L / R376L, E77V / V352L, L82Q, L82Q / I127L,L82Q / I127L / W144Y / T235S / R290V, L82Q / I127L / W144Y / A266V, L82Q / I127L / W144Y / A266V / R290V,L82Q / I127L / W144Y / K378S, L82Q / I127L / T235S, L82Q / I127L / T235S / A266V / I358W, L82Q / I127L / R290Q, L82Q / I127L / R290Q / I358W, L82Q / I127L / R290V / K378S, L82Q / W144Y / A266V / R290Q / K378S, L82Q / W144Y / A266V / K378S, L82Q / H150Q / T235S / A266V, L82Q / T235S / A266V, L82Q / T235S / A266V / R290Q, L82Q / T235S / A266V / K378S / A383P, L82Q / T235S / R290V, L82Q / T235S / K378S, L82Q / A266V, L82Q / A266V / K378S, L82Q / R290V, L82Q / K378S,S113A / K270L / T370S / A384V, S113E, S113E / N252A, S113E / N252A / K270L / T370S, S113E / G336Q, A118S / A233L / Q276E / G334L, A118S / G334L, 1127L / T235S / A266V, I127L / A266V / R290Q / K378S, I127L / K378S / A383P, D143H, D143H / Q276E, D143H / Q276E / G334L, W144Y / T235S / R290Q, W144Y / A266V / R290Q, W144Y / R290V / I358W, A233L / G334L, A233L / R376L, T235S, T235S / A266V / R290Q / I358W / A383P, T235S / I358W, N252A / K329A / G336Q / T370S, N252A / A384V, R256G / T333L, R256G / L375S, V262L, V262L / Q276E, V262L / G334L, A266V / R290Q, A266V / K378S, K270L / T370S, D273V, Q276E / V352L, R290Q, R290V / K378S, T333L, G334L, G334L / R376L, G336Q / T370S, R341 G, I358W, T370S, L375S, and K378S.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 3402 and one or more residue differences as compared to SEQ ID NO: 3402, selected from 18, 18 / 77 / 118 / 169 / 233 / 238 / 256 / 266 / 270, 18 / 77 / 150 / 256 / 376, 18 / 118, 18 / 118 / 143 / 169 / 238 / 256 / 266 / 333, 18 / 118 / 150 / 169 / 184 / 238 / 256 / 270 / 378, 18 / 118 / 169, 18 / 118 / 233 / 238, 18 / 143, 18 / 143 / 256, 18 / 150 / 169, 18 / 150 / 266, 18 / 169 / 184 / 290 / 333 / 376, 18 / 169 / 233 / 333, 18 / 169 / 392, 18 / 184 / 270, 18 / 233 / 256 / 334, 18 / 256 / 266 / 290 / 378, 77 / 113 / 118 / 256 / 266 / 290, 77 / 118 / 290, 77 / 169 / 270 / 290, 77 / 233, 77 / 233 / 256, 77 / 238, 113 / 118 / 169 / 184 / 256 / 376, 113 / 169, 113 / 233, 118, 118 / 143 / 238, 118 / 233 / 238 / 256 / 378, 118 / 233 / 238 / 270, 143 / 169 / 233 / 270, 143 / 233 / 266 / 290 / 376, 143 / 238 / 270 / 290 / 378, 143 / 256 / 334, 143 / 290, 169, 169 / 256 / 266, 183 / 270 / 333, 233 / 238, 233 / 238 / 376, 233 / 392, 238 / 256, 256 / 290 / 303, 270, 290, 376, and 378. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 3402 and one or more residue differences as compared to SEQ ID NO: 3402, selected from 18L, 18L / 77V / 118S / 169T / 233L / 238V / 256R / 266V / 270P, 18L / 77V / 150N / 256R / 376L, 18L / 118S, 18L / 118S / 143H / 169T / 238V / 256R / 266V / 333T, 18L / 118S / 150N / 169T / 184A / 238V / 256R / 270P / 378S, 18L / 118S / 169T, 18L / 118S / 233L / 238V, 18L / 143H, 18L / 143H / 256R, 18L / 150N / 169T, 18L / 150N / 266V, 18L / 169T / 184A / 290K / 333T / 376L, 18L / 169T / 233L / 333T, 18L / 169T / 392P, 18L / 184A / 270P, 18L / 233L / 256R / 334L, 18L / 256R / 266V / 290K / 378S, 77V / 113E / 118S / 256R / 266V / 290K, 77V / 118S / 290K, 77V / 169T / 270P / 290K, 77V / 233L, 77V / 233L / 256R, 77V / 238V, 113E / 118S / 169T / 184A / 256R / 376L, 113E / 169T, 113E / 233L, 118S, 118S / 143H / 238V, 118S / 233L / 238V / 256R / 378S, 118S / 233L / 238V / 270P, 143H / 169T / 233L / 270P, 143H / 233L / 266V / 290K / 376L, 143H / 238V / 270P / 290K / 378S, 143H / 256R / 334L, 143H / 290K, 169T, 169T / 256R / 266V, 183L / 270P / 333T, 233U238V, 233L / 238V / 376L, 233L / 392P, 238V / 256R, 256R / 290K / 303G, 270P, 290K, 376L, and 378S. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity toreference sequence SEQ ID NO: 3402 and one or more residue differences as compared to SEQ ID NO: 3402, selected from A18L, A18L / E77V / A118S / C169T / A233L / I238V / G256R / A266V / K270P, A18L / E77V / H150N / G256R / R376L, A18L / A118S,A18L / A118S / D143H / C169T / I238V / G256R / A266V / L333T,A18L / A118S / H150N / C169T / C184A / I238V / G256R / K270P / K378S, A18L / A118S / C169T, A18L / A118S / A233L / I238V, A18L / D143H, A18L / D143H / G256R, A18L / H150N / C169T, A18L / H150N / A266V, A18L / C169T / C184A / R290K / L333T / R376L, A18L / C169T / A233L / L333T, A18L / C169T / C392P, A18L / C184A / K270P, A18L / A233L / G256R / G334L,A18L / G256R / A266V / R290K / K378S, E77V / S113E / A118S / G256R / A266V / R290K, E77V / A118S / R290K, E77V / C169T / K270P / R290K, E77V / A233L, E77V / A233L / G256R, E77V / I238V,S113E / A118S / C169T / C184A / G256R / R376L, S113E / C169T, S113E / A233L, A118S, A118S / D143H / I238V, A118S / A233L / I238V / G256R / K378S, A118S / A233L / I238V / K270P, D143H / C169T / A233L / K270P, D143H / A233L / A266V / R290K / R376L, D143H / I238V / K270P / R290K / K378S, D143H / G256R / G334L, D143H / R290K, C169T, C169T / G256R / A266V, V183L / K270P / L333T, A233L / I238V, A233L / I238V / R376L, A233L / C392P, I238V / G256R, G256R / R290K / E303G, K270P, R290K, R376L, and K378S.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 3402 and one or more residue differences as compared to SEQ ID NO: 3402, selected from 18, 18 / 77 / 118 / 169 / 266 / 334 / 376, 18 / 77 / 150 / 256 / 376, 18 / 77 / 256 / 290 / 334, 18 / 113 / 238, 18 / 118, 18 / 118 / 143 / 169 / 238 / 256 / 266 / 333, 18 / 118 / 143 / 169 / 376, 18 / 118 / 169, 18 / 118 / 233 / 238, 18 / 118 / 266, 18 / 143, 18 / 143 / 169 / 233 / 238 / 358, 18 / 143 / 233 / 238 / 290 / 376, 18 / 143 / 256, 18 / 143 / 290, 18 / 143 / 333 / 334 / 378, 18 / 150 / 169, 18 / 150 / 266, 18 / 169 / 184 / 290 / 333 / 376, 18 / 169 / 233 / 333, 18 / 169 / 238 / 270 / 334 / 376, 18 / 169 / 392, 18 / 233 / 256 / 334, 18 / 233 / 266 / 290 / 376, 18 / 256 / 266 / 290 / 378, 18 / 266 / 270 / 378, 18 / 378, 77 / 113 / 121 / 143 / 233 / 334, 77 / 118 / 169 / 233 / 266 / 270, 77 / 118 / 238 / 376, 77 / 118 / 290, 77 / 169 / 270 / 290, 77 / 233, 77 / 238, 113 / 238 / 376, 118, 118 / 143 / 238, 118 / 143 / 392, 118 / 169 / 184 / 233 / 266 / 270 / 333, 118 / 233 / 238 / 256 / 378, 118 / 256 / 290 / 333 / 334 / 376 / 378, 118 / 256 / 334 / 376, 118 / 266 / 290, 143 / 169 / 233 / 270, 143 / 233 / 266 / 290 / 376, 143 / 238 / 270 / 290 / 378, 143 / 256 / 334, 143 / 290, 143 / 364, 150 / 233 / 333, 169, 169 / 256 / 266, 183 / 270 / 333, 233 / 238, 233 / 238 / 376, 233 / 392, 238 / 256, 238 / 266 / 270 / 378, 256 / 266 / 334, 266 / 270 / 376, 266 / 333, and 378. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 3402 and one or more residue differences as compared to SEQ ID NO: 3402, selected from 18L, 18L / 77V / 118S / 169T / 266V / 334L / 376L, 18L / 77V / 150N / 256R / 376L, 18L / 77V / 256R / 290K / 334L, 18L / 113E / 238V, 18L / 118S, 18L / 118S / 143H / 169T / 238V / 256R / 266V / 333T, 18L / 118S / 143H / 169T / 376L, 18L / 118S / 169T, 18L / 118S / 233L / 238V, 18L / 118S / 266V, 18L / 143H, 18L / 143H / 169T / 233L / 238V / 358W, 18L / 143H / 233L / 238V / 290K / 376L, 18L / 143H / 256R, 18L / 143H / 290K, 18L / 143H / 333T / 334L / 378S, 18L / 150N / 169T, 18L / 150N / 266V, 18L / 169T / 184A / 290K / 333T / 376L, 18L / 169T / 233L / 333T, 18L / 169T / 238V / 270P / 334L / 376L, 18L / 169T / 392P, 18L / 233L / 256R / 334L, 18L / 233L / 266V / 290K / 376L, 18L / 256R / 266V / 290K / 378S, 18L / 266V / 270P / 378S, 18L / 378S, 77V / 113E / 121 C / 143H / 233L / 334L,77V / 118S / 169T / 233L / 266 V / 270 P, 77V / 118S / 238V / 376L, 77V / 118S / 290K, 77V / 169T / 270P / 290K, 77V / 233L, 77V / 238V, 113E / 238V / 376L, 118S, 118S / 143H / 238V, 118S / 143H / 392P, 118S / 169T / 184A / 233L / 266V / 270P / 333T, 118S / 233L / 238V / 256R / 378S,118S / 256R / 290K / 333T / 334L / 376L / 378S, 118S / 256R / 334L / 376L, 118S / 266V / 290K, 143H / 169T / 233L / 270P, 143H / 233L / 266V / 290K / 376L, 143H / 238V / 270P / 290K / 378S, 143H / 256R / 334L, 143H / 290K, 143H / 364R, 150N / 233L / 333T, 169T, 169T / 256R / 266V, 183L / 270P / 333T, 233L / 238V, 233L / 238V / 376L, 233L / 392P, 238V / 256R, 238V / 266V / 270P / 378S, 256R / 266V / 334L, 266V / 270P / 376L, 266V / 333T, and 378S. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 3402 and one or more residue differences as compared to SEQ ID NO: 3402, selected from A18L, A18L / E77V / A118S / C169T / A266V / G334L / R376L, A18L / E77V / H150N / G256R / R376L, A18L / E77V / G256R / R290K / G334L, A18L / S113E / I238V, A18L / A118S,A18L / A118S / D143H / C169T / I238V / G256R / A266V / L333T, A18UA118S / D143H / C169T / R376L,A18L / A118S / C169T, A18L / A118S / A233L / I238V, A18L / A118S / A266V, A18L / D143H, A18L / D143H / C169T / A233L / I238V / I358W, A18L / D143H / A233L / I238V / R290K / R376L, A18L / D143H / G256R, A18L / D143H / R290K, A18L / D143H / L333T / G334L / K378S, A18L / H150N / C169T, A18L / H150N / A266V, A18L / C169T / C184A / R290K / L333T / R376L, A18L / C169T / A233L / L333T, A18L / C169T / I238V / K270P / G334L / R376L, A18L / C169T / C392P, A18L / A233L / G256R / G334L,A18L / A233L / A266V / R290K / R376L, A18L / G256R / A266V / R290K / K378S, A18L / A266V / K270P / K378S, A18L / K378S, E77V / S113E / W121 C / D143H / A233L / G334L, E77V / A118S / C169T / A233L / A266V / K270P, E77V / A118S / I238V / R376L, E77V / A118S / R290K, E77V / C169T / K270P / R290K, E77V / A233L, E77V / I238V, S113E / I238V / R376L, A118S, A118S / D143H / I238V, A118S / D143H / C392P,A118S / C169T / C184A / A233L / A266V / K270P / L333T, A118S / A233L / I238V / G256R / K378S,A118S / G256R / R290K / L333T / G334L / R376L / K378S, A118S / G256R / G334L / R376L, A118S / A266V / R290K, D143H / C169T / A233L / K270P, D143H / A233L / A266V / R290K / R376L, D143H / I238V / K270P / R290K / K378S, D143H / G256R / G334L, D143H / R290K, D143H / Q364R, H150N / A233L / L333T, C169T, C169T / G256R / A266V, V183L / K270P / L333T, A233L / I238V, A233L / I238V / R376L, A233L / C392P, I238V / G256R, I238V / A266V / K270P / K378S, G256R / A266V / G334L, A266V / K270P / R376L, A266V / L333T, and K378S.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 3662 and one or more residue differences as compared to SEQ ID NO: 3662, selected from 77 / 156, 77 / 156 / 270, 77 / 184 / 244, 77 / 184 / 333, 77 / 244 / 270, 77 / 270, 143, 143 / 169 / 238 / 257 / 344, 143 / 169 / 255, 143 / 169 / 255 / 257, 143 / 169 / 343, 143 / 255, 143 / 255 / 257, 143 / 255 / 344, 143 / 257, 156, 156 / 270, 169 / 238, 169 / 238 / 255 / 257 / 344, 169 / 257 / 344, 169 / 344, 184, 184 / 244 / 324, 238, 238 / 255 / 344, 244 / 270, 257 / 344, and 270. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQID NO: 3662 and one or more residue differences as compared to SEQ ID NO: 3662, selected from 77E / 156L, 77E / 156L / 270P, 77E / 184A / 244K, 77E / 184A / 333T, 77E / 244K / 270P, 77E / 270P, 77E / 270R, 143E, 143E / 169T / 238V / 257R / 344K, 143E / 169T / 255R, 143E / 169T / 255R / 257R, 143E / 255R, 143E / 255R / 257R, 143E / 255R / 344K, 143E / 257R, 143H / 169T / 343R, 156L, 156L / 270R, 169T / 238V, 169T / 238V / 255R / 257R / 344M, 169T / 257R / 344M, 169T / 344K, 169T / 344M, 184A, 184A / 244K / 324M, 238V, 238V / 255R / 344M, 244K / 270P, 257R / 344K, and 270P. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 3662 and one or more residue differences as compared to SEQ ID NO: 3662, selected from V77E / E156L, V77E / E156L / K270P, V77E / C184A / V244K, V77E / C184A / L333T, V77E / V244K / K270P, V77E / K270P, V77E / K270R, D143E, D143E / C169T / I238V / K257R / E344K, D143E / C169T / A255R, D143E / C169T / A255R / K257R, D143E / A255R, D143E / A255R / K257R, D143E / A255R / E344K, D143E / K257R, D143H / C169T / E343R, E156L, E156L / K270R, C169T / I238V, C169T / I238V / A255R / K257R / E344M, C169T / K257R / E344M, C169T / E344K, C169T / E344M, C184A, C184A / V244K / V324M, I238V, I238V / A255R / E344M, V244K / K270P, K257R / E344K, and K270P.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 3662 and one or more residue differences as compared to SEQ ID NO: 3662, selected from 77, 77 / 156 / 184 / 244 / 324, 77 / 184 / 270, 143, 143 / 238, 143 / 238 / 255, 143 / 238 / 255 / 343, 143 / 238 / 255 / 392, 143 / 238 / 344, 143 / 255, 143 / 257 / 392, 143 / 344, 169, 169 / 238 / 255 / 344, 169 / 238 / 344, 169 / 255, 169 / 343, 169 / 392, 238 / 344, 255, 255 / 257, 255 / 257 / 392, 257, and 344. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 3662 and one or more residue differences as compared to SEQ ID NO: 3662, selected from 77E, 77E / 156L / 184A / 244K / 324M, 77E / 184A / 270P, 143E / 238V, 143 E / 238 V / 255 R / 343R, 143E / 238V / 255R / 392P, 143E / 238V / 344M, 143E / 257R / 392P, 143E / 344M, 143H, 143H / 238V / 255R, 143H / 255R, 169T, 169T / 238V / 255R / 344M, 169T / 238V / 344M, 169T / 255R, 169T / 343R, 169T / 392P, 238V / 344M, 255R, 255R / 257R, 255R / 257R / 392P, 257R, and 344M. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 3662 and one or more residue differences as compared to SEQ ID NO: 3662, selected from V77E, V77E / E156L / C184A / V244K / V324M, V77E / C184A / K270P, D143E / I238V, D143E / I238V / A255R / E343R, D143E / I238V / A255R / C392P, D143E / I238V / E344M, D143E / K257R / C392P, D143E / E344M, D143H, D143H / I238V / A255R, D143H / A255R, C169T, C169T / I238V / A255R / E344M, C169T / I238V / E344M, C169T / A255R, C169T / E343R, C169T / C392P, I238V / E344M, A255R, A255R / K257R, A255R / K257R / C392P, K257R, and E344M.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%,89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 3788 and one or more residue differences as compared to SEQ ID NO: 3788, selected from 20, 22, 26, 42, 43, 44, 45, 50, 51 , 52, 53, 54, 55, 60, 62, 64, 65, 95, 97, 99, 103, 104, 111 , 112, 115, 115 / 143 / 169, 115 / 169 / 343, 115 / 343, 116, 116 / 343, 143, 143 / 169, 154 / 439, 169, 169 / 324, 169 / 324 / 343, 169 / 343, 186, 234, 236, 238, 238 / 246, 239, 239 / 244, 241 , 242, 243, 244, 250, 251 , 254, 255, 260, 283, 284, 295, 296, 298 / 326, 319, 322, 323, 324, 324 / 343, 326, 328, 329, 333, 334, 338, 340, 341 , 343, 344, 347, 366, 434, 439, 442, 443, 444, 445, 446, and 447. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 3788 and one or more residue differences as compared to SEQ ID NO: 3788, selected from 20R, 22F, 22L, 22N, 22R, 22V, 22W, 26R, 42G, 42P, 43L, 43P, 43V, 44G, 44P, 44V, 45A, 45E, 45G, 45L, 45M, 45Q, 45R, 45W, 50R, 51 Q, 51 R, 52K, 52P, 52R, 52T, 53A, 53H, 53R, 53S, 54R, 54S, 54T, 54V, 55G, 55M, 55S, 55T, 55V, 60L, 62G, 64A, 64D, 64E, 64G, 64R, 64S, 64T, 64V, 65A, 65M, 95G, 95R, 95V, 97A, 97C, 99G, 99R, 103D, 103F, 103G, 103R, 103S, 103T, 103V, 104G, 104T, 111 V, 112R, 115E, 115E / 169T / 343R, 115G, 115L, 115P, 115Q, 115Q / 143E / 169T, 115Q / 169T / 343R, 115Q / 343R, 115R, 115T, 115W, 116E / 343R, 116S, 143E, 143E / 169T, 154E / 439S, 169T, 169T / 324M, 169T / 324M / 343R, 169T / 343R, 186A, 186P, 186Q, 186S, 234W, 236T, 236V, 236Y, 238L / 246G, 238V, 239D, 239K / 244F, 239L, 241 R, 242M, 243E, 243G, 243L, 243Q, 243W, 244C, 244G, 244T, 250A, 250S, 251 A, 2511, 251 L, 251 S, 254R, 254T, 255C, 255K, 255L, 255M, 255R, 260A, 260C, 260I, 260T, 283G, 283S, 283T, 284A, 284G, 284R, 295F, 295I, 295V, 296H, 298I / 326A, 319E, 319K, 319R, 319W, 322G, 322S, 322T, 323H, 323T, 323V, 324M, 324M / 343R, 326A, 326L, 326Q, 328Y, 329G, 329T, 329W, 333M, 333Q, 333R, 333W, 333Y, 334E, 334W, 338A, 340A, 341 K, 343G, 343M, 343R, 343V, 344G, 344H, 344K, 344M, 344Q, 344S, 344T, 344V, 347R, 366S, 434Y, 439A, 439G, 439S, 439Y, 442G, 442H, 442Q, 443C, 443I, 443P, 443S, 444Q, 445G, 445L, 445M, 445T, 445V, 446F, 446G, 446R, 447L, 447N, and 447T. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 3788 and one or more residue differences as compared to SEQ ID NO: 3788, selected from G20R, P22F, P22L, P22N, P22R, P22V, P22W, K26R, S42G, S42P, I43L, I43P, I43V, T44G, T44P, T44V, V45A, V45E, V45G, V45L, V45M, V45Q, V45R, V45W, S50R, K51 Q, K51 R, A52K, A52P, A52R, A52T, M53A, M53H, M53R, M53S, Q54R, Q54S, Q54T, Q54V, E55G, E55M, E55S, E55T, E55V, F60L, R62G, P64A, P64D, P64E, P64G, P64R, P64S, P64T, P64V, N65A, N65M, T95G, T95R, T95V, S97A, S97C, S99G, S99R, Q103D, Q103F, Q103G, Q103R, Q103S, Q103T, Q103V, V104G, V104T, L111 V, G112R, K115E, K115E / C169T / E343R, K115G, K115L, K115P, K115Q, K115Q / D143E / C169T, K115Q / C169T / E343R, K115Q / E343R, K115R, K115T, K115W, D116E / E343R, D116S, D143E, D143E / C169T, K154E / R439S, C169T, C169T / V324M, C169T / V324M / E343R, C169T / E343R, R186A, R186P, R186Q, R186S, Y234W, I236T, I236V, I236Y, I238L / S246G, I238V, P239D, P239K / V244F, P239L, K241 R, G242M, V243E, V243G, V243L, V243Q, V243W, V244C, V244G, V244T, R250A, R250S, K251 A, K251 I, K251 L, K251 S, P254R, P254T, A255C, A255K, A255L, A255M, A255R, L260A, L260C, L260I, L260T, A283G, A283S, A283T, E284A, E284G, E284R, L295F, L295I, L295V, K296H, V298I / E326A, T319E, T319K, T319R,T319W, W322G, W322S, W322T, R323H, R323T, R323V, V324M, V324M / E343R, E326A, E326L, E326Q, H328Y, K329G, K329T, K329W, L333M, L333Q, L333R, L333W, L333Y, G334E, G334W, E338A, L340A, R341 K, E343G, E343M, E343R, E343V, E344G, E344H, E344K, E344M, E344Q, E344S, E344T, E344V, N347R, R366S, F434Y, R439A, R439G, R439S, R439Y, I442G, I442H, I442Q, A443C, A443I, A443P, A443S, S444Q, W445G, W445L, W445M, W445T, W445V, E446F, E446G, E446R, A447L, A447N, and A447T.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 4150 and one or more residue differences as compared to SEQ ID NO: 4150, selected from 26, 26 / 236, 26 / 236 / 260 / 333, 26 / 236 / 295, 26 / 260, 26 / 260 / 295, 26 / 260 / 295 / 333, 26 / 295 / 319, 26 / 333, 64, 64 / 244, 64 / 251 / 260 / 347, 64 / 260 / 295 / 333, 64 / 260 / 347, 103 / 115 / 255, 103 / 115 / 343, 103 / 115 / 343 / 344, 103 / 143 / 255 / 284 / 343 / 344, 103 / 169 / 343 / 344, 103 / 255 / 284, 103 / 343 / 344, 115, 115 / 143 / 255, 115 / 143 / 255 / 343, 115 / 143 / 284, 115 / 143 / 343, 115 / 143 / 344, 115 / 169, 115 / 169 / 343, 115 / 255, 115 / 255 / 344, 115 / 284, 115 / 284 / 343 / 344, 115 / 284 / 344, 115 / 343, 115 / 343 / 344, 115 / 344, 143, 143 / 169 / 255 / 284 / 343 / 344, 143 / 169 / 343 / 344, 143 / 255, 143 / 255 / 284 / 343 / 344, 143 / 255 / 343, 143 / 255 / 343 / 344, 143 / 255 / 344, 143 / 343 / 344, 169, 169 / 255, 169 / 255 / 284, 169 / 255 / 343, 169 / 284, 169 / 343, 169 / 344, 236, 236 / 244 / 333, 236 / 260, 236 / 347, 244, 244 / 251 , 244 / 251 / 319, 244 / 260, 244 / 295, 244 / 347, 251 , 251 / 260 / 295 / 347, 251 / 333, 255, 255 / 284, 255 / 343, 255 / 343 / 344, 255 / 344, 259, 260, 260 / 295, 260 / 333, 284, 284 / 343 / 344, 284 / 344, 295, 295 / 333, 333, 343, 343 / 344, 344, and 347. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 4150 and one or more residue differences as compared to SEQ ID NO: 4150, selected from 26R, 26R / 236T, 26R / 236T / 260I / 333Y, 26R / 236T / 295I, 26R / 260I, 26R / 260I / 295I, 26R / 260I / 295I / 333Y, 26R / 295I / 319K, 26R / 333Y, 64A, 64A / 244T, 64A / 251 A / 260I / 347R, 64A / 260I / 295I / 333Y, 64A / 260I / 347R, 103G / 115E / 255K, 103G / 115E / 343M, 103G / 115E / 343M / 344G, 103G / 143E / 255K / 284G / 343M / 344G, 103G / 169T / 343M / 344G, 103G / 255K / 284A, 103G / 343M / 344G, 115E, 115E / 143E / 255K, 115E / 143E / 344G, 115E / 169T, 115E / 255K, 115E / 284A / 344G, 115E / 284G / 343M / 344G, 115E / 343M, 115E / 344G, 115T, 115T / 143E / 255K / 343M, 115T / 143E / 284G, 115T / 143E / 343M, 115T / 169T, 115T / 169T / 343M, 115T / 255K, 115T / 255K / 344G, 115T / 284G, 115T / 343M / 344G, 115T / 344G, 143E, 143E / 169T / 255K / 284G / 343M / 344G, 143E / 169T / 343M / 344G, 143E / 255K, 143E / 255K / 284A / 343M / 344G, 143E / 255K / 343M, 143E / 255K / 343M / 344G, 143E / 255K / 344G, 143E / 343M / 344G, 169T, 169T / 255K, 169T / 255K / 284G, 169T / 255K / 343M, 169T / 284A, 169T / 284G, 169T / 343M, 169T / 344G, 236T, 236T / 244T / 333Y, 236T / 260I, 236T / 347R, 244T, 244T / 251A, 244T / 251 A / 319K, 244T / 260I, 244T / 295I, 244T / 347R, 251 A, 251 A / 260I / 295I / 347R, 251A / 333Y, 255K, 255K / 284G, 255K / 343M, 255K / 343M / 344G, 255K / 344G, 259I, 260I, 260I / 295I, 260I / 333Y, 284A, 284A / 343M / 344G, 284G, 284G / 344G, 295I, 295I / 333Y, 333Y, 343M, 343M / 344G, 344G, and 347R. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO:4150 and one or more residue differences as compared to SEQ ID NO: 4150, selected from K26R, K26R / I236T, K26R / I236T / L260I / L333Y, K26R / I236T / L295I, K26R / L260I, K26R / L260I / L295I, K26R / L260I / L295I / L333Y, K26R / L295I / T319K, K26R / L333Y, P64A, P64A / V244T,P64A / K251 A / L260I / N347R, P64A / L260I / L295I / L333Y, P64A / L260I / N347R, Q103G / K115E / A255K, Q103G / K115E / E343M, Q103G / K115E / E343M / E344G, Q103G / D143E / A255K / E284G / E343M / E344G, Q103G / C169T / E343M / E344G, Q103G / A255K / E284A, Q103G / E343M / E344G, K115E,K115E / D143E / A255K, K115E / D143E / E344G, K115E / C169T, K115E / A255K, K115E / E284A / E344G, K115E / E284G / E343M / E344G, K115E / E343M, K115E / E344G, K115T, K115T / D143E / A255K / E343M, K115T / D143E / E284G, K115T / D143E / E343M, K115T / C169T, K115T / C169T / E343M, K115T / A255K, K115T / A255K / E344G, K115T / E284G, K115T / E343M / E344G, K115T / E344G, D143E,D143E / C169T / A255K / E284G / E343M / E344G, D143E / C169T / E343M / E344G, D143E / A255K, D143E / A255K / E284A / E343M / E344G, D143E / A255K / E343M, D143E / A255K / E343M / E344G, D143E / A255K / E344G, D143E / E343M / E344G, C169T, C169T / A255K, C169T / A255K / E284G, C169T / A255K / E343M, C169T / E284A, C169T / E284G, C169T / E343M, C169T / E344G, I236T, I236T / V244T / L333Y, I236T / L260I, I236T / N347R, V244T, V244T / K251A, V244T / K251 A / T319K, V244T / L260I, V244T / L295I, V244T / N347R, K251 A, K251 A / L260I / L295I / N347R, K251A / L333Y, A255K, A255K / E284G, A255K / E343M, A255K / E343M / E344G, A255K / E344G, S259I, L260I, L260I / L295I, L260I / L333Y, E284A, E284A / E343M / E344G, E284G, E284G / E344G, L295I, L295I / L333Y, L333Y, E343M, E343M / E344G, E344G, and N347R.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 4278 and one or more residue differences as compared to SEQ ID NO: 4278, selected from 115, 115 / 117, 115 / 117 / 118, 115 / 117 / 118 / 152, 115 / 117 / 118 / 169, 115 / 117 / 118 / 246, 115 / 117 / 152, 115 / 117 / 152 / 169, 115 / 117 / 152 / 169 / 246, 115 / 117 / 152 / 246, 115 / 117 / 169, 115 / 117 / 246, 115 / 118, 115 / 118 / 152, 115 / 118 / 152 / 169, 115 / 118 / 152 / 169 / 246, 115 / 118 / 169, 115 / 118 / 246, 115 / 152, 115 / 152 / 169, 115 / 152 / 169 / 246, 115 / 152 / 246, 115 / 152 / 246 / 374, 115 / 169, 115 / 169 / 246, 115 / 246, 117, 117 / 118, 117 / 118 / 152, 117 / 118 / 152 / 169, 117 / 118 / 152 / 246, 117 / 118 / 169, 117 / 118 / 246, 117 / 152, 117 / 152 / 169, 117 / 152 / 246 / 288, 117 / 169, 117 / 246, 118 / 246, 152, 152 / 169, 152 / 169 / 246, 152 / 246, 169, 169 / 246, and 246. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 4278 and one or more residue differences as compared to SEQ ID NO: 4278, selected from 115E, 115E / 117A, 115E / 117A / 118T, 115E / 117A / 118T / 152R, 115E / 117A / 118T / 169T, 115E / 117A / 118T / 246D, 115E / 117A / 152R, 115E / 117A / 152R / 169T, 115E / 117A / 152R / 169T / 246D, 115E / 117A / 152R / 246D, 115E / 117A / 169T, 115E / 117A / 246D, 115E / 118T, 115E / 118T / 152R, 115E / 118T / 152R / 169T, 115E / 118T / 152R / 169T / 246D, 115E / 118T / 169T, 115E / 118T / 246D, 115E / 152R, 115E / 152R / 169T, 115E / 152R / 169T / 246D, 115E / 152R / 246D, 115E / 152R / 246D / 374C, 115E / 169T, 115E / 169T / 246D, 115E / 246D, 117A, 117A / 118T, 117A / 118T / 152R, 117A / 118T / 152R / 169T, 117A / 118T / 152R / 246D, 117A / 118T / 169T, 117A / 118T / 246D, 117A / 152R, 117A / 152R / 169T, 117A / 152R / 246D / 288G, 117A / 169T,117A / 246D, 118T / 246D, 152R, 152R / 169T, 152R / 169T / 246D, 152R / 246D, 169T, 169T / 246D, and 246D. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 4278 and one or more residue differences as compared to SEQ ID NO: 4278, selected from K115E, K115E / K117A, K115E / K117A / S118T, K115E / K117A / S118T / T152R, K115E / K117A / S118T / C169T, K115E / K117A / S118T / S246D, K115E / K117A / T152R, K115E / K117A / T152R / C169T,K115E / K117A / T152R / C169T / S246D, K115E / K117A / T152R / S246D, K115E / K117A / C169T, K115E / K117A / S246D, K115E / S118T, K115E / S118T / T152R, K115E / S118T / T152R / C169T, K115E / S118T / T152R / C169T / S246D, K115E / S118T / C169T, K115E / S118T / S246D, K115E / T152R, K115E / T152R / C169T, K115E / T152R / C169T / S246D, K115E / T152R / S246D,K115E / T152R / S246D / Y374C, K115E / C169T, K115E / C169T / S246D, K115E / S246D, K117A,K117A / S118T, K117A / S118T / T152R, K117A / S118T / T152R / C169T, K117A / S118T / T152R / S246D, K117A / S118T / C169T, K117A / S118T / S246D, K117A / T152R, K117A / T152R / C169T,K117A / T152R / S246D / S288G, K117A / C169T, K117A / S246D, S118T / S246D, T152R, T152R / C169T, T152R / C169T / S246D, T152R / S246D, C169T, C169T / S246D, and S246D.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 4278 and one or more residue differences as compared to SEQ ID NO: 4278, selected from 73, 101 , 114, 115, 118, 143, 145, 150, 151 , 152, 153, 154, 156, 157, 160, 161 , 163, 166, 167, 168, 169, 171 , 172, 190, 191 , 192, 195, 213, 214, 218, 219, 223, 225, 233, 233 / 343, 234, 235, 236, 237, 238, 239, 239 / 250, 243, 244, 249, 251 , 254, 255, 257, 258, 259, 261 , 262, 262 / 283, 277 / 292, 283, 284, 286, 289, 292, 295, and 296. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 4278 and one or more residue differences as compared to SEQ ID NO: 4278, selected from 73L, 73R, 101 S, 114S, 115A, 115Q, 115R, 115S, 118R, 143E, 143L, 143Q, 143V, 145Y, 1501, 150L, 150M, 150T, 151 A, 151 D, 151 F, 151 H, 151 R, 152L, 153G, 154P, 154S, 154Y, 156D, 156L, 156Q, 156W, 157L,157P, 157R, 160R, 160T, 160V, 161 R, 163K, 163L, 163Q, 163R, 166T, 167Q, 167R, 167T, 168A, 168N,168V, 169L, 169V, 1711, 171 L, 171 M, 172L, 172R, 172V, 190F, 191 H, 191 L, 191 V, 192E, 192G, 192R, 192S, 195L, 195M, 195T, 195V, 213R, 214L, 214V, 218P, 219R, 223A, 223D, 223Q, 225V, 233F, 233G,233L, 233Q, 233R, 233T / 343G, 233W, 234L, 234V, 235C, 235D, 235E, 235S, 236M, 236T, 237Q, 238L,238V, 239A, 239E, 239N / 250H, 239T, 243I, 243Q, 243V, 244A, 244F, 244L, 244S, 244T, 244W, 249G, 249I, 249L, 249R, 249S, 249T, 251 H, 251 Q, 254A, 254E, 255G, 255S, 255W, 257L, 257T, 258V, 259L, 259N, 261 A, 261 K, 261 R, 261 T, 261 V, 262I / 283V, 262L, 277S / 292V, 283G, 283T, 284I, 284L, 284R, 284S, 286E, 286G, 286I, 286L, 286V, 289L, 292E, 292G, 295V, 296C, and 296R. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 4278 and one or more residue differences ascompared to SEQ ID NO: 4278, selected from Q73L, Q73R, K101 S, V1 14S, K1 15A, K1 15Q, K1 15R, K1 15S, S1 18R, D143E, D143L, D143Q, D143V, W145Y, H150I, H150L, H150M, H150T, P151 A, P151 D, P151 F, P151 H, P151 R, T152L, D153G, K154P, K154S, K154Y, E156D, E156L, E156Q, E156W, K157L, K157P, K157R, A160R, A160T, A160V, K161 R, A163K, A163L, A163Q, A163R, S166T, H167Q, H167R, H167T, K168A, K168N, K168V, C169L, C169V, Q171 1, Q171 L, Q171 M, 1172L, 1172R, 1172V, 1190F, Q191 H, Q191 L, Q191 V, A192E, A192G, A192R, A192S, Q195L, Q195M, Q195T, Q195V, K213R, I214L, 1214V, E218P, L219R, E223A, E223D, E223Q, L225V, A233F, A233G, A233L, A233Q, A233R, A233T / E343G, A233W, Y234L, Y234V, T235C, T235D, T235E, T235S, I236M, I236T, D237Q, I238L, I238V, P239A, P239E, P239N / R250H, P239T, G243I, G243Q, G243V, V244A, V244F, V244L, V244S, V244T, V244W, K249G, K249I, K249L, K249R, K249S, K249T, K251 H, K251 Q, P254A, P254E, A255G, A255S, A255W, K257L, K257T, A258V, S259L, S259N, S261 A, S261 K, S261 R, S261 T, S261 V, V262I / A283V, V262L, P277S / M292V, A283G, A283T, E284I, E284L, E284R, E284S, T286E, T286G, T286I, T286L, T286V, P289L, M292E, M292G, I295V, K296C, and K296R.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 4642 and one or more residue differences as compared to SEQ ID NO: 4642, selected from 93, 1 13, 1 13 / 1 14, 1 13 / 1 15, 1 13 / 143, 1 15 / 1 18, 1 15 / 177, 1 18 / 143 / 163 / 168 / 195 / 255 / 315, 1 18 / 143 / 255, 1 18 / 163 / 168 / 214 / 255 / 315, 1 18 / 168 / 214 / 315, 1 18 / 255, 124, 126, 129, 134, 143 / 154 / 255, 143 / 156 / 168 / 216 / 255, 143 / 163, 143 / 163 / 255, 143 / 168, 143 / 168 / 169 / 255 / 315, 143 / 255, 148, 156 / 214 / 315, 159, 163, 163 / 168, 163 / 168 / 169, 163 / 168 / 169 / 255 / 315, 163 / 168 / 169 / 315, 163 / 168 / 195 / 214 / 255 / 315, 163 / 214, 163 / 255, 163 / 255 / 315, 168 / 315, 186, 194, 197, 206, 207, 208, 210, 213, 255 / 315, 277, 293, 294, 315, 317, 320, 322, 328, 329, 330, 333, 352, 366, 392, and 445. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 4642 and one or more residue differences as compared to SEQ ID NO: 4642, selected from 93A, 93F, 93G, 93H, 93M, 93S, 93T, 1 13C / 1 15K, 1 13G / 1 15K, 1 13S, 1 13S / 1 14G, 1 13S / 1 15E, 1 13S / 143K, 113S / 143R, 1 15K / 1 18C, 1 15K / 1 18G, 1 15R / 177V, 1 18T / 143Q / 163V / 168N / 195E / 255K / 315L, 1 18T / 143Q / 255K, 1 18T / 163V / 168N / 214L / 255K / 315L, 1 18T / 168N / 214L / 315L, 1 18T / 255K, 124T, 126V, 129M, 134R, 134V, 143Q / 154L / 255K, 143Q / 156D / 168N / 216G / 255K, 143Q / 163V, 143Q / 163V / 255K, 143Q / 168N, 143Q / 168N / 169L / 255K / 315L, 143Q / 255K, 148L, 148Q, 148R, 156D / 214L / 315L, 159T, 163V, 163V / 168M, 163V / 168M / 169L, 163V / 168M / 169T, 163V / 168M / 169V, 163V / 168M / 195E / 214L / 255K / 315L, 163V / 168N / 169T / 255K / 315L, 163V / 168N / 169V / 315L, 163V / 214L, 163V / 255K, 163V / 255K / 315L, 168N / 315L, 186K, 194M, 197L, 206L, 206M, 207M, 208G, 21 OR, 213R, 255K / 315L, 277D, 277G, 277S, 277W, 293G, 293P, 293R, 293S, 293T, 294G, 294R, 294V, 315L, 317A, 317C, 317G, 317L, 317Q, 317S, 320C, 320I, 320V, 320Y, 322F, 328F, 329L, 329S, 330S, 330T, 333G, 333Q, 333R, 352G, 352I, 366A, 366K, 392A, 445R, and 445Y. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 2 and one or more residue differencesas compared to SEQ ID NO: 4642, selected from D93A, D93F, D93G, D93H, D93M, D93S, D93T, E1 13C / Q1 15K, E1 13G / Q1 15K, E1 13S, E1 13S / V1 14G, E1 13S / Q1 15E, E1 13S / D143K, E1 13S / D143R, Q115K / S1 18C, Q1 15K / S1 18G, Q1 15R / M177V, S1 18T / D143Q / A163V / K168N / Q195E / A255K / I315L, S1 18T / D143Q / A255K, S1 18T / A163V / K168N / I214L / A255K / I315L, S1 18T / K168N / I214L / I315L, S1 18T / A255K, H124T, T126V, L129M, T134R, T134V, D143Q / K154L / A255K, D143Q / E156D / K168N / D216G / A255K, D143Q / A163V, D143Q / A163V / A255K, D143Q / K168N, D143Q / K168N / C169L / A255K / I315L, D143Q / A255K, P148L, P148Q, P148R, E156D / I214L / I315L, S159T, A163V, A163V / K168M, A163V / K168M / C169L, A163V / K168M / C169T, A163V / K168M / C169V, A163V / K168M / Q195E / I214L / A255K / I315L, A163V / K168N / C169T / A255K / I315L, A163V / K168N / C169V / I315L, A163V / I214L, A163V / A255K, A163V / A255K / I315L, K168N / I315L, R186K, L194M, K197L, V206L, V206M, R207M, A208G, H210R, K213R, A255K / I315L, P277D, P277G, P277S, P277W, E293G, E293P, E293R, E293S, E293T, P294G, P294R, P294V, 1315L, M317A, M317C, M317G, M317L, M317Q, M317S, A320C, A320I, A320V, A320Y, W322F, H328F, K329L, K329S, A330S, A330T, L333G, L333Q, L333R, V352G, V352I, R366A, R366K, C392A, W445R, and W445Y.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 4642 and one or more residue differences as compared to SEQ ID NO: 4642, selected from 127, 128, 129, 140, 180, 182, 194, 277, 293, 294, 299, 300, 317, 319, 320, 324, 333, 356, 366, and 392. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 4642 and one or more residue differences as compared to SEQ ID NO: 4642, selected from 127L, 128V, 129M, 140V, 180L, 182Q, 182T, 194M, 277A, 277G, 277H, 277S, 293G, 293K, 293P, 293S, 294G, 294L, 294Q, 299T, 300T, 317A, 317F, 317I, 317S, 319A, 320I, 320Y, 324I, 333D, 333T, 356G, 366K, 392H, and 392M. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 4642 and one or more residue differences as compared to SEQ ID NO: 4642, selected from I127L, L128V, L129M, 1140V, V180L, S182Q, S182T, L194M, P277A, P277G, P277H, P277S, E293G, E293K, E293P, E293S, P294G, P294L, P294Q, L299T, L300T, M317A, M317F, M317I, M317S, T319A, A320I, A320Y, V324I, L333D, L333T, A356G, R366K, C392H, and C392M.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 4984 and one or more residue differences as compared to SEQ ID NO: 4984, selected from 1 13, 1 17, 1 18, 151 , 191 , 196, and 205. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 4984 and one or more residue differences as compared to SEQ ID NO: 4984,selected from 113A, 113C, 113K, 113R, 113S, 113T, 117R, 118A, 118K, 118R, 151 A, 151C, 151 E, 151 F, 151G, 151 H, 1511, 151 K, 151 L, 151 M, 151 N, 1510, 151 R, 151 S, 151T, 151V, 151W, 151 Y, 191G, 191 K, 191 L, 191 M, 191 R, 191S, 191V, 196A, 196F, 196G, 196H, 1961, 196K, 196L, 196M, 196R, 196S, 196V, 196W, 196Y, and 205T. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93% 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 4984 and one or more residue differences as compared to SEQ ID NO: 4984, selected from E113A, E113C, E113K, E113R, E113S, E113T, K117R, S118A, S118K, S118R, P151A, P151C, P151E, P151F, P151G, P151H, P151I, P151K, P151L, P151M, P151N, P151Q, P151R, P151S, P151T, P151V, P151W, P151Y, Q191G, Q191K, Q191 L, Q191M, Q191R, Q191S, Q191V, D196A, D196F, D196G, D196H, D196I, D196K, D196L, D196M, D196R, D196S, D196V, D196W, D196Y, and V205T.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 4984 and one or more residue differences as compared to SEQ ID NO: 4984, selected from 114 / 115 / 143, 114 / 115 / 143 / 233, 114 / 115 / 233, 114 / 115 / 343, 114 / 143, 114 / 143 / 151 / 233 / 238 / 261, 114 / 143 / 213 / 261 / 343, 114 / 143 / 233, 114 / 143 / 233 / 238, 114 / 151 / 261 / 343, 114 / 213 / 238, 114 / 343, 115, 115 / 143, 115 / 143 / 151 / 154 / 213 / 261, 115 / 143 / 151 / 154 / 233 / 343, 115 / 143 / 151 / 233 / 261 , 115 / 143 / 151 / 233 / 343, 115 / 143 / 154 / 213 / 233 / 238 / 261 , 115 / 143 / 213, 115 / 143 / 233, 115 / 143 / 343, 115 / 151 / 154 / 233, 115 / 151 / 261, 115 / 213 / 233, 115 / 213 / 233 / 238, 115 / 213 / 261, 115 / 233 / 238, 115 / 238, 115 / 343, 115 / 348, 143, 143 / 151 / 154, 143 / 151 / 154 / 213, 143 / 154, 143 / 213 / 233, 143 / 213 / 233 / 238 / 261 / 343, 143 / 213 / 261 / 343, 143 / 233 / 261, 143 / 261, 151 / 154, 151 / 154 / 213 / 343, 151 / 154 / 343, 151 / 213, 151 / 213 / 233, 213 / 233 / 343, and 233 / 238 / 343. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 4984 and one or more residue differences as compared to SEQ ID NO: 4984, selected from 114G / 115E / 143E / 233T, 114G / 115E / 143V, 114G / 115E / 233T, 114G / 115E / 343G, 114G / 143E / 233Q, 114G / 143L / 233F / 238V, 114G / 143V,114G / 143V / 151 R / 233T / 238 V / 261 R, 114G / 143V / 213R / 261 R / 343G, 114G / 151 R / 261 R / 343G, 114G / 213R / 238V, 114G / 343G, 115E, 115E / 143E, 115E / 143E / 213R, 115E / 143L / 151 R / 154S / 213R / 261 R, 115E / 143L / 151 R / 154S / 233Q / 343G, 115E / 143L / 151 R / 233F / 343G,115E / 143L / 154S / 213R / 233Q / 238V / 261 R, 115E / 143L / 233Q, 115E / 143L / 233R, 115E / 143L / 343G, 115E / 143V / 151 R / 233F / 261 R, 115E / 151 R / 154S / 233F, 115E / 151 R / 261 R, 115E / 213R / 233F / 238V, 115E / 213R / 233R, 115E / 213R / 261 R, 115E / 233F / 238V, 115E / 238V, 115E / 343G, 115E / 348F, 143E, 143E / 151 R / 154S, 143E / 151 R / 154S / 213R, 143E / 213R / 233F / 238V / 261 R / 343G, 143E / 233F / 261 R, 143L / 213R / 233R, 143L / 261R, 143V / 154S, 143V / 213R / 233R, 143V / 213R / 261 R / 343G, 151 R / 154S, 151 R / 154S / 213R / 343G, 151 R / 154S / 343G, 151 R / 213R, 151 R / 213R / 233F, 213R / 233R / 343G, and 233T / 238V / 343G. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 4984 and oneor more residue differences as compared to SEQ ID NO: 4984, selected fromV114G / Q115E / D143E / A233T, V114G / Q115E / D143V, V114G / Q115E / A233T, V114G / Q115E / E343G, V114G / D143E / A233Q, V114G / D143L / A233F / I238V, V114G / D143V,V114G / D143V / P151 R / A233T / I238V / S261 R, V114G / D143V / K213R / S261 R / E343G,V114G / P151 R / S261 R / E343G, V114G / K213R / I238V, V114G / E343G, Q115E, Q115E / D143E, Q115E / D143E / K213R, Q115E / D143L / P151 R / K154S / K213R / S261 R,Q115E / D143L / P151 R / K154S / A233Q / E343G, Q115E / D143L / P151 R / A233F / E343G,Q115E / D143L / K154S / K213R / A233Q / I238V / S261 R, Q115E / D143L / A233Q, Q115E / D143L / A233R, Q115E / D143L / E343G, Q115E / D143V / P151 R / A233F / S261 R, Q115E / P151 R / K154S / A233F,Q115E / P151 R / S261 R, Q115E / K213R / A233F / I238V, Q115E / K213R / A233R, Q115E / K213R / S261 R, Q115E / A233F / I238V, Q115E / I238V, Q115E / E343G, Q115E / L348F, D143E, D143E / P151 R / K154S, D143E / P151 R / K154S / K213R, D143E / K213R / A233F / I238V / S261 R / E343G, D143E / A233F / S261 R, D143L / K213R / A233R, D143L / S261 R, D143V / K154S, D143V / K213R / A233R,D143V / K213R / S261 R / E343G, P151 R / K154S, P151 R / K154S / K213R / E343G, P151 R / K154S / E343G, P151 R / K213R, P151 R / K213R / A233F, K213R / A233R / E343G, and A233T / I238V / E343G.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 5196 and one or more residue differences as compared to SEQ ID NO: 5196, selected from 117, 117 / 118 / 143 / 168 / 169 / 392, 117 / 118 / 143 / 392, 117 / 118 / 168 / 169, 117 / 118 / 168 / 169 / 392, 117 / 143 / 168 / 169 / 170 / 392, 117 / 143 / 168 / 169 / 392, 117 / 168 / 169, 117 / 168 / 169 / 392, 117 / 168 / 392, 117 / 169, 117 / 392, 118 / 143, 118 / 143 / 168 / 169, 118 / 143 / 169, 118 / 143 / 392, 118 / 168, 118 / 168 / 392, 118 / 169 / 392, 118 / 392, 143 / 168 / 169 / 392, 143 / 169, 143 / 392, 168 / 169, 168 / 169 / 392, 168 / 392, 169 / 392, and 392. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 5196 and one or more residue differences as compared to SEQ ID NO: 5196, selected from 117R, 117R / 118R / 143Q / 168K / 169T / 392A, 117R / 118R / 143Q / 392A, 117R / 118R / 168K / 169M, 117R / 118R / 168K / 169T / 392A, 117R / 118R / 168K / 169T / 392P, 117R / 143Q / 168K / 169T / 170H / 392A, 117R / 143Q / 168K / 169T / 392A, 117R / 168K / 169T, 117R / 168K / 169T / 392P, 117R / 168K / 392A, 117R / 169T, 117R / 392P, 118R / 143Q, 118R / 143Q / 168K / 169T, 118R / 143Q / 169T, 118R / 143Q / 392A, 118R / 143Q / 392P, 118R / 168K, 118R / 168K / 392P, 118R / 169T / 392A, 118R / 392A, 118R / 392P, 143Q / 168K / 169T / 392P, 143Q / 169T, 143Q / 392P, 168K / 169T, 168K / 169T / 392A, 168K / 169T / 392P, 168K / 392A, 168K / 392P, 169T / 392S, 392A, and 392P. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 5196 and one or more residue differences as compared to SEQ ID NO: 5196, selected from K117R, K117R / S118R / D143Q / M168K / V169T / C392A, K117R / S118R / D143Q / C392A, K117R / S118R / M168K / V169M, K117R / S118R / M168K / V169T / C392A,K117R / S118R / M168K / V169T / C392P, K117R / D143Q / M168K / V169T / R170H / C392A,K1 17R / D143Q / M168K / V169T / C392A, K1 17R / M168K / V169T, K117R / M168K / V169T / C392P,K1 17R / M168K / C392A, K1 17R / V169T, K1 17R / C392P, S1 18R / D143Q, S1 18R / D143Q / M168K / V169T, S1 18R / D143Q / V169T, S1 18R / D143Q / C392A, S1 18R / D143Q / C392P, S1 18R / M168K, S1 18R / M168K / C392P, S1 18R / V169T / C392A, S1 18R / C392A, S1 18R / C392P, D143Q / M168K / V169T / C392P, D143Q / V169T, D143Q / C392P, M168K / V169T, M168K / V169T / C392A, M168K / V169T / C392P, M168K / C392A, M168K / C392P, V169T / C392S, C392A, and C392P.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 5286 and one or more residue differences as compared to SEQ ID NO: 5286, selected from 134, 134 / 182 / 194 / 294, 134 / 182 / 277 / 293, 134 / 182 / 293 / 320, 134 / 182 / 294, 134 / 194 / 277, 134 / 194 / 277 / 317, 134 / 194 / 293, 134 / 194 / 293 / 294, 134 / 194 / 293 / 317, 134 / 194 / 294, 134 / 194 / 317, 134 / 194 / 317 / 320, 134 / 317, 182, 182 / 194, 182 / 194 / 293 / 320, 182 / 293 / 294, 194, 194 / 277 / 293 / 317, 194 / 277 / 317 / 320, 194 / 277 / 320, 194 / 293 / 294 / 317, 194 / 293 / 320, 277, 293, 317, 317 / 320, and 320. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 5286 and one or more residue differences as compared to SEQ ID NO: 5286, selected from 134P, 134R / 182T / 277W / 293G, 134R / 182T / 294Q, 134R / 194M / 277W, 134R / 194M / 277W / 317S, 134R / 194M / 293P, 134R / 194M / 293P / 294Q, 134R / 194M / 293P / 317S,134R / 194M / 294Q, 134R / 194M / 317S, 134R / 194M / 317S / 320Y, 134R / 317S, 134V, 134V / 182T / 194M / 294Q, 134V / 182T / 293P / 320Y, 134V / 194M / 277W, 134V / 194M / 293P, 182T, 182T / 194M, 182T / 194M / 293G / 320Y, 182T / 293G / 294Q, 194M, 194M / 277W / 293G / 317S, 194M / 277W / 317S / 320Y, 194M / 277W / 320Y, 194M / 293P / 294Q / 317S, 194M / 293P / 320Y, 277W, 293P, 317S, 317S / 320Y, and 320Y. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 5286 and one or more residue differences as compared to SEQ ID NO: 5286, selected from T134P, T134R / S182T / P277W / E293G, T134R / S182T / P294Q, T134R / L194M / P277W, T134R / L194M / P277W / M317S, T134R / L194M / E293P, T134R / L194M / E293P / P294Q, T134R / L194M / E293P / M317S, T134R / L194M / P294Q, T134R / L194M / M317S, T134R / L194M / M317S / A320Y, T134R / M317S, T134V,T134V / S182T / L194M / P294Q, T134V / S182T / E293P / A320Y, T134V / L194M / P277W, T134V / L194M / E293P, S182T, S182T / L194M, S182T / L194M / E293G / A320Y, S182T / E293G / P294Q, L194M, L194M / P277W / E293G / M317S, L194M / P277W / M317S / A320Y, L194M / P277W / A320Y, L194M / E293P / P294Q / M317S, L194M / E293P / A320Y, P277W, E293P, M317S, M317S / A320Y, and A320Y.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 5412 and one or more residue differences as compared to SEQ ID NO: 5412, selected from 12 / 134, 13 / 134, 13 / 134 / 210, 22 / 134 / 152, 22 / 134 / 235 / 344, 64 / 93 / 134 / 235, 64 / 134 / 344 / 347,77 / 134, 79 / 134 / 210, 84 / 134, 86 / 134, 87 / 134, 89 / 134, 93 / 134 / 152 / 235 / 344, 93 / 134 / 235 / 344, 93 / 134 / 244 / 344, 93 / 134 / 344, 131 , 134 / 152 / 344, 134 / 175, 134 / 210 / 382, 134 / 210 / 402, 134 / 210 / 456, 134 / 210 / 478, 134 / 344, 134 / 377, 134 / 378, 134 / 380, 134 / 381 , 134 / 382, 134 / 384, 134 / 386, 134 / 387, 134 / 388, 134 / 390, 134 / 391 , 134 / 393, 134 / 395, 134 / 398, 134 / 401 , 134 / 402, 134 / 406, 134 / 407, 134 / 421 , 134 / 456, 134 / 457, 134 / 459, 134 / 460, 134 / 464, 134 / 467, 134 / 468, 134 / 470, 134 / 471 , 134 / 473, 134 / 473 / 474, 134 / 474, and 134 / 475. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 5412 and one or more residue differences as compared to SEQ ID NO: 5412, selected from 12N / 134R, 13C / 134T / 210R, 13L / 134R, 13L / 134T, 13R / 134R, 13S / 134R, 13T / 134R, 13V / 134R, 13V / 134T / 210R, 22R / 134R / 152P, 22R / 134R / 235S / 344T, 64R / 93T / 134R / 235S, 64R / 134R / 344Q / 347R, 77Y / 134R, 79V / 134T / 21 OR, 84V / 134R, 86S / 134R, 87G / 134R, 87M / 134R, 89C / 134R, 89L / 134R, 93T / 134R / 152P / 235S / 344T, 93T / 134R / 235S / 344T, 93T / 134R / 244L / 344T, 93T / 134R / 344T, 1311, 134R / 152P / 344Q, 134R / 175C, 134R / 210R / 402I, 134R / 344T, 134R / 377I, 134R / 378R, 134R / 378V, 134R / 380V, 134R / 381 D, 134R / 382G, 134R / 382K, 134R / 382W, 134R / 384F, 134R / 386W, 134R / 387G, 134R / 387I, 134R / 388F, 134R / 390V, 134R / 391 H, 134R / 391 R, 134R / 391T, 134R / 393T, 134R / 395I, 134R / 395M, 134R / 398S, 134R / 401 V, 134R / 402L, 134R / 402V, 134R / 406C, 134R / 407V, 134R / 421 V, 134R / 456D, 134R / 456T, 134R / 457M, 134R / 459L, 134R / 460E, 134R / 464A, 134R / 467L, 134R / 467R, 134R / 467T, 134R / 468S, 134R / 470L, 134R / 470P, 134R / 471 M, 134R / 471 S, 134R / 473D / 474I, 134R / 473G, 134R / 474T, 134R / 475G, 134T / 210R / 382R, 134T / 210R / 456C, 134T / 210R / 478D, and 134T / 380V. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 5412 and one or more residue differences as compared to SEQ ID NO: 5412, selected from E12N / P134R, H13C / P134T / H210R, H13L / P134R, H13L / P134T, H13R / P134R, H13S / P134R, H13T / P134R, H13V / P134R, H13V / P134T / H210R, P22R / P134R / T152P, P22R / P134R / T235S / E344T, P64R / D93T / P134R / T235S, P64R / P134R / E344Q / N347R, E77Y / P134R, T79V / P134T / H210R, Q84V / P134R, Y86S / P134R, P87G / P134R, P87M / P134R, I89C / P134R, I89L / P134R, D93T / P134R / T152P / T235S / E344T, D93T / P134R / T235S / E344T, D93T / P134R / V244L / E344T, D93T / P134R / E344T, G131 I, P134R / T152P / E344Q, P134R / D175C, P134R / H210R / K402I, P134R / E344T, P134R / K377I, P134R / K378R, P134R / K378V, P134R / L380V, P134R / V381 D, P134R / D382G, P134R / D382K, P134R / D382W, P134R / A384F, P134R / E386W, P134R / L387G, P134R / L387I, P134R / E388F, P134R / Q390V, P134R / S391 H, P134R / S391 R, P134R / S391T, P134R / A393T, P134R / V395I, P134R / V395M, P134R / E398S, P134R / W401 V, P134R / K402L, P134R / K402V, P134R / K406C, P134R / M407V, P134R / L421 V, P134R / Q456D, P134R / Q456T, P134R / S457M, P134R / V459L, P134R / V460E, P134R / V464A, P134R / E467L, P134R / E467R, P134R / E467T, P134R / M468S, P134R / A470L, P134R / A470P, P134R / D471 M, P134R / D471 S, P134R / E473D / L474I, P134R / E473G, P134R / L474T, P134R / L475G, P134T / H210R / D382R, P134T / H210R / Q456C, P134T / H210R / S478D, and P134T / L380V.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 5412 and one or more residue differences as compared to SEQ ID NO: 5412, selected from 73 / 127 / 134 / 160 / 172 / 198 / 290, 73 / 127 / 134 / 160 / 198 / 333, 73 / 127 / 134 / 160 / 210 / 266, 73 / 127 / 134 / 172 / 192 / 266 / 333, 73 / 127 / 134 / 172 / 198, 73 / 127 / 134 / 210 / 290, 73 / 127 / 134 / 210 / 333, 73 / 127 / 134 / 290 / 333, 73 / 127 / 134 / 333, 73 / 134, 73 / 134 / 160 / 172 / 192 / 290, 73 / 134 / 160 / 172 / 198 / 333, 73 / 134 / 160 / 172 / 290, 73 / 134 / 160 / 266, 73 / 134 / 160 / 333, 73 / 134 / 172, 73 / 134 / 172 / 210 / 333, 73 / 134 / 172 / 266 / 333, 73 / 134 / 198 / 210, 73 / 134 / 198 / 210 / 266, 73 / 134 / 210 / 333, 73 / 134 / 333, 73 / 134 / 366, 127 / 134, 127 / 134 / 160 / 172 / 192 / 333, 127 / 134 / 160 / 172 / 210 / 266 / 290, 127 / 134 / 172 / 198 / 266 / 333, 127 / 134 / 192 / 210, 127 / 134 / 198 / 210 / 290, 127 / 134 / 210 / 266 / 290, 127 / 134 / 210 / 333, 134, 134 / 160 / 172 / 333, 134 / 160 / 198 / 266 / 290, 134 / 172 / 192 / 290 / 333, 134 / 192 / 198, 134 / 198, 134 / 198 / 266 / 333, 134 / 210, 134 / 210 / 290, and 134 / 333. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 5412 and one or more residue differences as compared to SEQ ID NO: 5412, selected from 73K / 127L / 134R / 160G / 172R / 198Q / 290R, 73K / 127L / 134R / 160G / 198Q / 333T, 73K / 127L / 134R / 172R / 192S / 266A / 333T, 73K / 127L / 134R / 172R / 198Q, 73K / 127L / 134R / 333T, 73K / 127L / 134T / 160G / 210R / 266A, 73K / 127L / 134T / 210R / 290R, 73K / 127L / 134T / 210R / 333T, 73K / 127L / 134T / 290R / 333T, 73K / 134R, 73K / 134R / 160G / 172R / 290R, 73K / 134R / 160G / 333T, 73K / 134R / 172R, 73K / 134R / 172R / 210R / 333T, 73K / 134R / 172R / 266A / 333T, 73K / 134R / 333T, 73K / 134R / 366H, 73K / 134T / 160G / 172R / 192S / 290R, 73K / 134T / 160G / 172R / 198Q / 333T, 73K / 134T / 160G / 266A, 73K / 134T / 198Q / 210R, 73K / 134T / 198Q / 210R / 266A, 73K / 134T / 210R / 333T, 127L / 134R, 127L / 134R / 160G / 172R / 192S / 333T, 127L / 134R / 172R / 198Q / 266A / 333T, 127L / 134R / 210R / 266A / 290R, 127L / 134T, 127L / 134T / 160G / 172R / 210R / 266A / 290R, 127L / 134T / 192S / 210R, 127L / 134T / 198Q / 210R / 290R, 127L / 134T / 210R / 333T, 134R, 134R / 160G / 172R / 333T, 134R / 172R / 192S / 290R / 333T, 134R / 192S / 198Q, 134R / 198Q, 134R / 198Q / 266A / 333T, 134R / 333T, 134T / 160G / 198Q / 266A / 290R, 134T / 210R, and 134T / 210R / 290R.In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 5412 and one or more residue differences as compared to SEQ ID NO: 5412, selected from Q73K / I127L / P134R / A160G / I172R / R198Q / K290R, Q73K / I127L / P134R / A160G / R198Q / L333T, Q73K / I127L / P134R / I172R / A192S / V266A / L333T, Q73K / I127L / P134R / I172R / R198Q, Q73K / I127L / P134R / L333T, Q73K / I127L / P134T / A160G / H210R / V266A,Q73K / I127L / P134T / H210R / K290R, Q73K / I127L / P134T / H210R / L333T, Q73K / I127L / P134T / K290R / L333T, Q73K / P134R, Q73K / P134R / A160G / I172R / K290R, Q73K / P134R / A160G / L333T, Q73K / P134R / I172R, Q73K / P134R / I172R / H210R / L333T, Q73K / P134R / I172R / V266A / L333T, Q73K / P134R / L333T, Q73K / P134R / R366H, Q73K / P134T / A160G / I172R / A192S / K290R,Q73K / P134T / A160G / I172R / R198Q / L333T, Q73K / P134T / A160G / V266A, Q73K / P134T / R198Q / H21 OR,Q73K / P134T / R198Q / H210R / V266A, Q73K / P134T / H210R / L333T, 1127L / P134R, 1127L / P134R / A160G / I172R / A192S / L333T, 1127L / P134R / I172R / R198Q / V266A / L333T, 1127L / P134R / H210R / V266A / K290R, 1127L / P134T, 1127L / P134T / A160G / I172R / H210R / V266A / K290R, 1127L / P134T / A192S / H21 OR, I127L / P134T / R198Q / H210R / K290R, I127L / P134T / H210R / L333T, P134R, P134R / A160G / I172R / L333T, P134R / I172R / A192S / K290R / L333T, P134R / A192S / R198Q, P134R / R198Q, P134R / R198Q / V266A / L333T, P134R / L333T, P134T / A160G / R198Q / V266A / K290R, P134T / H210R, and P134T / H210R / K290R.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 5422 and one or more residue differences as compared to SEQ ID NO: 5422, selected from 13, 13 / 178 / 381 , 13 / 178 / 381 / 473, 13 / 178 / 457, 13 / 381 , 13 / 381 / 457, 13 / 402 / 457, 13 / 457, 178, 178 / 381 / 457, 178 / 381 / 457 / 473, 178 / 402 / 457, 178 / 473, 381 , 381 / 393 / 457, 381 / 457 / 473, 381 / 473, and 393 / 457. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 5422 and one or more residue differences as compared to SEQ ID NO: 5422, selected from 13L, 13L / 178R / 381 D / 473G, 13L / 178R / 381W, 13L / 178R / 457M, 13L / 381 D / 457M, 13L / 381W, 13L / 402L / 457M, 13L / 457M, 178R, 178R / 381 D / 457M / 473G, 178R / 381 W / 457M, 178R / 402L / 457M, 178R / 473G, 381 D / 457M / 473G, 381 W, 381 W / 393T / 457M, 381 W / 473G, and 393T / 457M. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 5422 and one or more residue differences as compared to SEQ ID NO: 5422, selected from H13L, H13L / S178R / V381 D / E473G, H13L / S178R / V381 W, H13L / S178R / S457M, H13L / V381 D / S457M, H13L / V381W, H13L / K402L / S457M, H13L / S457M, S178R,S178R / V381 D / S457M / E473G, S178R / V381 W / S457M, S178R / K402L / S457M, S178R / E473G, V381 D / S457M / E473G, V381W, V381 W / A393T / S457M, V381W / E473G, and A393T / S457M.In some embodiments, the engineered transposase polypeptide with one or more improved properties comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 972 and one or more residue differences as compared to SEQ ID NO: 972, selected from 38 / 73 / 113 / 115 / 117 / 118 / 127 / 134 / 152 / 154 / 160 / 163 / 164 / 168 / 169 / 172 / 184 / 192 / 194 / 198 / 220 / 243 / 260 / 266 / 27 0 / 295 / 317 / 333 / 392 / 423, 38 / 73 / 113 / 115 / 117 / 118 / 127 / 134 / 152 / 154 / 160 / 163 / 164 / 168 / 169 / 172 / 184 / 192 / 198 / 220 / 243 / 260 / 266 / 270 / 29 5 / 317 / 333 / 392 / 423, 38 / 73 / 113 / 115 / 117 / 118 / 127 / 134 / 152 / 154 / 160 / 163 / 164 / 168 / 169 / 172 / 184 / 192 / 198 / 220 / 243 / 260 / 266 / 270 / 29 5 / 317 / 333 / 392 / 423 / 457, 38 / 73 / 113 / 115 / 117 / 118 / 127 / 152 / 154 / 160 / 163 / 164 / 168 / 169 / 172 / 184 / 192 / 198 / 220 / 243 / 260 / 266 / 270 / 295 / 31 7 / 333 / 392 / 423,38 / 73 / 1 13 / 1 15 / 1 18 / 127 / 152 / 154 / 160 / 162 / 164 / 168 / 172 / 184 / 192 / 198 / 220 / 243 / 260 / 266 / 270 / 295 / 317 / 333 / 42 3,38 / 73 / 1 13 / 1 15 / 1 18 / 127 / 152 / 154 / 160 / 163 / 164 / 168 / 169 / 172 / 184 / 192 / 198 / 220 / 243 / 260 / 266 / 270 / 295 / 317 / 33 3 / 423,38 / 73 / 1 13 / 1 15 / 1 18 / 127 / 152 / 154 / 160 / 164 / 168 / 172 / 184 / 189 / 192 / 198 / 220 / 243 / 260 / 266 / 270 / 295 / 317 / 333 / 423,38 / 73 / 1 13 / 1 15 / 1 18 / 127 / 152 / 154 / 160 / 164 / 168 / 172 / 184 / 192 / 198 / 210 / 220 / 243 / 260 / 266 / 270 / 295 / 317 / 333 / 42 3, 38 / 73 / 1 13 / 1 15 / 1 18 / 127 / 152 / 154 / 160 / 164 / 168 / 172 / 184 / 192 / 198 / 220 / 243 / 260 / 266 / 270 / 295 / 317 / 333 / 423, 38 / 73 / 1 13 / 1 18 / 127 / 152 / 154 / 160 / 164 / 168 / 172 / 184 / 192 / 198 / 220 / 243 / 260 / 266 / 270 / 295 / 317 / 333 / 423, 38 / 73 / 1 13 / 1 18 / 127 / 152 / 154 / 160 / 164 / 168 / 172 / 184 / 192 / 198 / 220 / 243 / 266 / 270 / 317 / 333 / 423,38 / 73 / 1 13 / 1 18 / 127 / 152 / 154 / 160 / 164 / 168 / 172 / 184 / 192 / 198 / 220 / 266 / 270 / 317 / 333 / 423, and38 / 1 13 / 127 / 152 / 154 / 160 / 164 / 168 / 172 / 198 / 220 / 290 / 317 / 423. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 972 and one or more residue differences as compared to SEQ ID NO: 972, selected from38F / 73Q / 1 13E / 1 15E / 1 17R / 118R / 1271 / 134R / 152T / 154K / 160A / 163V / 164R / 168K / 169T / 1721 / 184A / 192A / 19 8R / 220R / 243G / 260I / 266V / 270P / 295I / 317M / 333L / 392A / 423D,38F / 73Q / 1 13E / 1 15E / 1 17R / 1 18R / 1271 / 134R / 152T / 154K / 160A / 163V / 164R / 168K / 169T / 1721 / 184A / 192A / 19 8R / 220R / 243G / 260I / 266V / 270P / 295I / 317M / 333L / 392A / 423D / 457M,38F / 73Q / 1 13E / 1 15E / 1 17R / 1 18R / 1271 / 134V / 152T / 154K / 160A / 163V / 164R / 168K / 169T / 1721 / 184A / 192A / 194 M / 198R / 220R / 243G / 260I / 266V / 270P / 295I / 317M / 333L / 392A / 423D,38F / 73Q / 1 13E / 1 15E / 1 17R / 1 18R / 1271 / 152T / 154K / 160A / 163V / 164R / 168K / 169T / 1721 / 184A / 192A / 198R / 22 0R / 243G / 260I / 266V / 270P / 295I / 317M / 333L / 392A / 423D,38F / 73Q / 1 13E / 1 15E / 1 18S / 1271 / 152T / 154K / 160A / 163V / 164R / 168M / 169V / 1721 / 184A / 192A / 198R / 220R / 24 3G / 260I / 266V / 270P / 295I / 317M / 333L / 423D,38F / 73Q / 1 13E / 1 15Q / 1 18S / 1271 / 152T / 154K / 160A / 162L / 164R / 168K / 1721 / 184A / 192A / 198R / 220R / 243G / 26 0I / 266V / 270P / 295I / 317M / 333L / 423D,38F / 73Q / 1 13E / 1 15Q / 1 18S / 1271 / 152T / 154K / 160A / 163V / 164R / 168M / 169V / 1721 / 184A / 192A / 198R / 220R / 24 3G / 260I / 266V / 270P / 295I / 317M / 333L / 423D,38F / 73Q / 1 13E / 1 15Q / 1 18S / 1271 / 152T / 154K / 160A / 164R / 168K / 1721 / 184A / 189S / 192A / 198R / 220R / 243G / 26 0I / 266V / 270P / 295I / 317M / 333L / 423D,38F / 73Q / 1 13E / 1 15Q / 1 18S / 1271 / 152T / 154K / 160A / 164R / 168K / 1721 / 184A / 192A / 198R / 210R / 220R / 243G / 26 0I / 266V / 270P / 295I / 317M / 333L / 423D,38F / 73Q / 1 13E / 1 15Q / 1 18S / 1271 / 152T / 154K / 160A / 164R / 168K / 1721 / 184A / 192A / 198R / 220R / 243G / 260I / 266 V / 270 P / 2951 / 317M / 333L / 423D,38F / 73Q / 1 13E / 1 18S / 1271 / 152T / 154K / 160A / 164R / 168K / 1721 / 184A / 192A / 198R / 220R / 243G / 260I / 266V / 270 P / 2951 / 317M / 333L / 423D,38F / 73Q / 1 13E / 1 18S / 1271 / 152T / 154K / 160A / 164R / 168K / 1721 / 184A / 192A / 198 R / 220R / 243G / 266 V / 270 P / 31 7M / 333L / 423D,38F / 73Q / 1 13E / 1 18S / 1271 / 152T / 154K / 160A / 164R / 168K / 1721 / 184A / 192A / 198R / 220R / 266V / 270P / 317M / 33 3L / 423D, and 38F / 1 13E / 1271 / 152T / 154K / 160A / 164R / 168K / 1721 / 198R / 220R / 290R / 317M / 423D. In some embodiments, the engineered transposase polypeptide comprises an amino acid sequence having at least 60%, 70%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91 %, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more sequence identity to reference sequence SEQ ID NO: 972 and one or more residue differences as compared to SEQ ID NO: 972, selected fromC38F / K73Q / S1 13E / K1 15E / K1 17R / A1 18R / L127I / T134R / E152T / A154K / G160A / A163V / D164R / F168K / C16 9T / R172I / C184A / S192A / Q198R / P220R / V243G / L260I / A266V / K270P / L295I / R317M / T333L / C392A / W423D, C38F / K73Q / S1 13E / K1 15E / K1 17R / A1 18R / L127I / T134R / E152T / A154K / G160A / A163V / D164R / F168K / C16 9T / R172I / C184A / S192A / Q198R / P220R / V243G / L260I / A266V / K270P / L295I / R317M / T333L / C392A / W423D / S457M,C38F / K73Q / S1 13E / K1 15E / K1 17R / A1 18R / L127I / T134V / E152T / A154K / G160A / A163V / D164R / F168K / C16 9T / R172I / C184A / S192A / L194M / Q198R / P220R / V243G / L260I / A266V / K270P / L295I / R317M / T333L / C392A / W423D,C38F / K73Q / S1 13E / K1 15E / K1 17R / A1 18R / L127I / E152T / A154K / G160A / A163V / D164R / F168K / C169T / R17 2I / C184A / S192A / Q198R / P220R / V243G / L260I / A266V / K270P / L295I / R317M / T333L / C392A / W423D, C38F / K73Q / S1 13E / K1 15E / A1 18S / L127I / E152T / A154K / G160A / A163V / D164R / F168M / C169V / R172I / C184 A / S192A / Q198R / P220R / V243G / L260I / A266V / K270P / L295I / R317M / T333L / W423D,C38F / K73Q / S1 13E / K1 15Q / A1 18S / L127I / E152T / A154K / G160A / W162L / D164R / F168K / R172I / C184A / S192 A / Q198R / P220R / V243G / L260I / A266V / K270P / L295I / R317M / T333L / W423D,C38F / K73Q / S1 13E / K1 15Q / A1 18S / L127I / E152T / A154K / G160A / A163V / D164R / F168M / C169V / R172I / C184 A / S192A / Q198R / P220R / V243G / L260I / A266V / K270P / L295I / R317M / T333L / W423D,C38F / K73Q / S1 13E / K1 15Q / A1 18S / L127I / E152T / A154K / G160A / D164R / F168K / R172I / C184A / D189S / S192 A / Q198R / P220R / V243G / L260I / A266V / K270P / L295I / R317M / T333L / W423D,C38F / K73Q / S1 13E / K1 15Q / A1 18S / L127I / E152T / A154K / G160A / D164R / F168K / R172I / C184A / S192A / Q198 R / H210R / P220R / V243G / L260I / A266V / K270P / L295I / R317M / T333L / W423D,C38F / K73Q / S1 13E / K1 15Q / A1 18S / L127I / E152T / A154K / G160A / D164R / F168K / R172I / C184A / S192A / Q198 R / P220 R / V243G / L2601 / A266 V / K270 P / L2951 / R317M / T333L / W423 D ,C38F / K73Q / S1 13E / A1 18S / L127I / E152T / A154K / G160A / D164R / F168K / R172I / C184A / S192A / Q198R / P220 R / V243G / L2601 / A266 V / K270 P / L2951 / R317M / T333L / W423 D ,C38F / K73Q / S1 13E / A1 18S / L127I / E152T / A154K / G160A / D164R / F168K / R172I / C184A / S192A / Q198R / P220 R / V243G / A266V / K270P / R317M / T333L / W423D,C38F / K73Q / S1 13E...

Claims

WHAT IS CLAIMED IS:1 . An engineered transposase comprising a polypeptide sequence having at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to the reference sequence of SEQ ID NOs: 2, 4, 156, 302, 338, 606, 972, 1082, 1218, 1580, 1598, 1600, 1604, 1720, 2368, 2426, 2540, 3402, 3662, 3788, 4150, 4278, 4642, 4984, 5196, 5286, 5412, 5422, and / or 5704, or a functional fragment thereof, wherein the engineered transposase comprises at least one substitution or substitution set in its polypeptide sequence, and wherein the amino acid positions of the polypeptide sequence are numbered with reference to SEQ ID NOs: 2, 4, 156, 302, 338, 606, 972, 1082, 1218, 1580, 1598, 1600, 1604, 1720, 2368, 2426, 2540, 3402, 3662, 3788, 4150, 4278, 4642, 4984, 5196, 5286, 5412, 5422, and / or 5704.

2. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 10, 13, 24, 35, 39, 44, 46, 51 , 54, 55, 62 / 361 , 72, 75, 82, 89, 92, 107, 128, 137, 164, 167, 177 / 359, 191 , 192, 193, 194, 201 , 217, 281 , 289, 291 , 291 / 340, 305, 309, 310, 317, 331 , 337, 340, 346, 352, 353, 357, 358, 359, 362, 371 , 396, 414 / 435, 415, 428, 437, 443, 447, and 448, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 2.

3. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 4, 5, 6, 8, 10, 10 / 317, 10 / 317 / 358, 10 / 317 / 359, 10 / 346, 10 / 346 / 359, 10 / 358, 10 / 359, 12, 12 / 373, 13, 15, 16, 18, 19, 26, 30, 30 / 154, 31 , 34, 35, 37, 38, 68, 70, 73, 74, 93, 106, 118, 126, 134, 135, 143, 146, 154 / 453, 163, 182, 207 / 286, 208, 226, 237, 246, 247, 259, 262, 263, 264, 280, 282, 286, 287, 288, 289, 291 , 292, 293, 298, 305, 309, 310, 313, 317, 317 / 346, 317 / 359, 317 / 445, 320, 346, 351 , 355, 356, 358, 359, 360, 368, 369, 410, 411 , 422, 426, 427, 428, 446, 448, 453, and 454, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 4.

4. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 18 / 73 / 74 / 259, 18 / 73 / 286 / 289 / 411 ,18 / 74 / 118 / 259 / 286 / 289 / 411 , 18 / 74 / 259, 18 / 74 / 259 / 286 / 289, 18 / 74 / 259 / 286 / 411 , 18 / 118 / 259, 18 / 259 / 286 / 289 / 411 , 18 / 259 / 289 / 411 , 18 / 289 / 411 , 73 / 74 / 259 / 286 / 289, 73 / 74 / 259 / 289, 73 / 259 / 286, 74 / 259, 74 / 259 / 289, 74 / 286 / 289 / 411 , 259, 259 / 286 / 289, and 289, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 156.

5. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 16 / 19 / 355 / 427,16 / 37 / 146 / 286 / 287 / 289 / 310 / 313 / 355, 16 / 146 / 226 / 287 / 289 / 355, 19 / 37 / 146 / 286 / 287 / 310 / 355,19 / 135 / 286 / 287 / 289 / 355, 19 / 182 / 355, 37 / 70 / 226 / 310 / 313 / 355, 37 / 74 / 310 / 355 / 427,37 / 135 / 146 / 182 / 226 / 286 / 287 / 355 / 368, 37 / 146 / 226 / 289 / 427, 37 / 146 / 287 / 289 / 427, 38, 41 , 42, 43, 44, 45, 47, 48, 50, 53, 54, 55, 70 / 74 / 226 / 286 / 287 / 289 / 368 / 427, 70 / 74 / 355 / 368, 70 / 135 / 146 / 150 / 182 / 286 / 289 / 313 / 427, 74 / 310 / 313 / 355, 74 / 310 / 355, 74 / 355 / 427, 101 , 103, 1 17, 135 / 226 / 286 / 289 / 355 / 368, 135 / 355, 146, 146 / 150 / 287 / 289 / 310 / 355, 146 / 182 / 355, 146 / 226 / 286 / 355, 146 / 286 / 287 / 289 / 355 / 427, 146 / 289, 146 / 427, 150 / 226 / 310 / 313, 182 / 226 / 355, 182 / 355, 244, 247, 249, 251 , 287 / 355 / 427, 300, 317 / 41 1 , 355, 393, 397, 398, 399, 401 , 405, 407, 408, 409, 41 1 , 413, 426, 429, 431 , 439, 444, 446, 450, 453, 454, 457, 459, 460, 464, 466, 467, 469, 470, 471 , 472, 474, 475, 476, and 478, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 302.

6. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 16, 16 / 313, 16 / 409 / 41 1 / 427,37 / 150 / 408, 37 / 313 / 317 / 408 / 41 1 , 37 / 317 / 405 / 408, 37 / 317 / 405 / 426, 37 / 405 / 409 / 427, 150, 150 / 313 / 405 / 41 1 , 150 / 405 / 406, 150 / 408, 150 / 408 / 41 1 , 226 / 405, 317 / 460, and 408 / 426, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 338.

7. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 12, 13, 18, 19, 22, 26, 31 / 262, 34, 60, 79, 80, 81 , 82, 92, 106, 107, 1 13, 1 15, 132, 134, 136, 140, 143, 152, 160, 163, 167, 168, 175, 179, 183,195, 199, 201 , 202, 215, 216, 218, 220, 223, 226, 233, 235, 236, 237, 243, 255, 259, 261 , 262, 266, 268, 270, 273, 282, 290, 292, 293, 309, 319, 320, 326, 343, 346, 365, 370, 374, 380, 382, 386, 389, 390, 408, 412, 414, 415, 424, and 442, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 606.

8. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 2, 12, 13, 15, 16, 18, 19, 22, 26, 30, 31 / 262, 34, 51 , 60, 69, 79, 82, 84, 87, 88, 106, 107, 1 13, 1 14, 1 15, 128, 132, 134, 135, 136, 140, 143,146, 150, 152, 160, 163, 167, 168, 175, 179, 183, 195, 198, 199, 201 , 215, 217, 218, 219, 220, 223, 224,226, 227, 231 , 233, 235, 236, 243, 254, 255, 259, 261 , 266, 267, 268, 270, 272, 273, 275, 277, 279, 282,287, 289, 290, 291 , 292, 293, 304, 307, 309, 319, 320, 343, 344, 346, 355, 365, 370, 373, 380, 382, 386,389, 390, 408, 412, 414, 415, 418, 423, 424, and 468, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 606.

9. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 44 / 45 / 47 / 408 / 453, 44 / 45 / 47 / 453, 44 / 45 / 47 / 453 / 454, 44 / 45 / 103 / 453 / 457, 44 / 45 / 453 / 454 / 457, 44 / 47 / 103 / 393 / 408 / 453 / 454,44 / 47 / 103 / 453 / 454 / 457, 44 / 47 / 393 / 408 / 453 / 457, 44 / 47 / 393 / 453 / 457, 44 / 103 / 408 / 453 / 454, 45 / 47 / 103 / 393 / 453 / 454 / 457, 45 / 47 / 393 / 453 / 454, 45 / 47 / 408 / 423 / 453 / 457, 45 / 47 / 453, 45 / 103 / 393 / 453 / 454, 45 / 453 / 454, 47 / 103 / 408 / 453 / 454, 47 / 393 / 408 / 453 / 454 / 457, 47 / 393 / 453 / 454, 47 / 408 / 453 / 457,53 / 54 / 244 / 450, 103 / 393 / 408 / 453 / 457, 244 / 450, 408 / 453 / 454, 446 / 450 / 464, and 453 / 457, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 606.

10. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 38 / 45 / 53, 38 / 45 / 53 / 54 / 476, 38 / 45 / 53 / 244, 38 / 45 / 53 / 470 / 472, 38 / 45 / 54 / 244, 38 / 45 / 54 / 470 / 474 / 478, 38 / 45 / 472 / 476 / 478, 38 / 45 / 476, 38 / 53 / 54, 38 / 53 / 54 / 244, 38 / 53 / 54 / 244 / 472 / 478, 38 / 53 / 472, 38 / 54 / 244 / 470 / 474 / 476 / 478, 38 / 244 / 470 / 472 / 474 / 478, 38 / 478, 45 / 53 / 54 / 244 / 470 / 474 / 476 / 478, 45 / 53 / 54 / 470 / 474 / 476, 45 / 53 / 54 / 472 / 478, 45 / 54 / 244 / 474 / 478, 53, 53 / 54 / 470 / 476 / 478, 53 / 244 / 474 / 478, 54, 54 / 244, 54 / 244 / 472 / 476, 54 / 472, 244, and 474 / 478, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 972.1 1 . The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 22 / 38 / 45 / 107 / 259 / 317 / 412, 22 / 38 / 45 / 259 / 317 / 412, 22 / 107, 22 / 107 / 317 / 412, 22 / 134, 22 / 134 / 220, 22 / 134 / 220 / 317, 22 / 134 / 317 / 412, 22 / 317, 38, 38 / 45 / 81 / 143 / 244, 38 / 53 / 243, 38 / 81 , 38 / 81 / 244, 38 / 107 / 134 / 220, 38 / 132, 38 / 220 / 317, 45 / 81 , 45 / 81 / 243, 45 / 81 / 243 / 343, 45 / 143 / 243 / 270 / 343, 45 / 244 / 270, 53 / 1 15 / 163 / 243 / 270 / 343, 53 / 143 / 243 / 270 / 343, 53 / 270, 64 / 134 / 220 / 317 / 412, 81 / 132 / 243, 107, 107 / 134 / 317, 1 15 / 244 / 270, 134 / 259, 143 / 243 / 270, 243, 243 / 270, 244, 244 / 270, 259, 270, and 317, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 972.

12. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 22 / 38 / 45 / 107 / 259 / 317 / 412, 22 / 38 / 45 / 259 / 317 / 412, 22 / 38 / 64 / 134 / 220, 22 / 45 / 220 / 259, 22 / 64 / 107 / 259 / 317 / 412, 22 / 64 / 134 / 220, 22 / 107, 22 / 134, 22 / 134 / 317, 22 / 134 / 317 / 412, 22 / 220, 22 / 220 / 412 / 415, 22 / 259 / 317, 22 / 259 / 317 / 412, 22 / 317, 38, 38 / 45 / 81 / 143 / 244, 38 / 45 / 143 / 270, 38 / 53 / 243, 38 / 81 , 38 / 81 / 143 / 243 / 270, 38 / 81 / 183 / 243 / 270, 38 / 81 / 243, 38 / 81 / 244, 38 / 107 / 134 / 220, 38 / 132, 38 / 220 / 317, 38 / 270 / 343, 45 / 81 , 45 / 81 / 132 / 243 / 270, 45 / 81 / 243, 45 / 143 / 243 / 270 / 343, 45 / 244 / 270, 53 / 143 / 243 / 270 / 343, 53 / 244 / 270, 53 / 270, 64, 64 / 107 / 134, 64 / 107 / 317, 64 / 134, 64 / 134 / 220 / 317 / 412, 81 / 132, 81 / 132 / 243, 107, 107 / 134 / 317, 107 / 412, 1 15 / 244 / 270, 143 / 243 / 270, 220 / 259, 243, 243 / 270, 244 / 270, 255, 259, and 270, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 972.

13. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 143, 143 / 175 / 272 / 273 / 415, 152 / 168 / 216 / 237, 152 / 168 / 423, 160, 160 / 201 / 272, 160 / 201 / 273 / 415, 160 / 226, 160 / 244 / 272 / 273 / 415, 160 / 415, 168, 168 / 187 / 216 / 255 / 423, 168 / 216 / 237, 168 / 255 / 423, 168 / 290 / 423, 187, 187 / 423, 199 / 201 / 226, 199 / 201 / 415, 201 , 216, 216 / 423, 226 / 244 / 415, 226 / 415, 237, 237 / 290, 244, 255, 272, 290, 290 / 423, 415, and 423, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 1082.

14. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 143, 143 / 160 / 226 / 415, 143 / 175 / 272 / 273 / 415, 152 / 168 / 216 / 237, 152 / 168 / 423, 160, 160 / 201 / 272, 160 / 201 / 273 / 415, 160 / 226, 160 / 244 / 272 / 273 / 415, 160 / 415, 168, 168 / 187 / 216 / 255 / 423, 168 / 216 / 237, 168 / 216 / 237 / 255 / 343, 168 / 255 / 423, 168 / 290 / 423, 187, 199 / 201 / 226, 199 / 201 / 415, 201 , 216, 216 / 423, 226 / 244 / 415, 226 / 415, 237, 237 / 290, 244, 255, 255 / 343, 272, 290, 290 / 423, 415, and 423, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 1082.

15. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 72, 90, 91 , 92, 93, 95, 122, 129, 141 , 142, 143, 144, 145, 150, 151 , 151 / 155, 152, 154, 155, 157, 160, 161 , 161 / 313, 164, 165, 168, 180, 181 , 184, 193, 197, 204, 205, 205 / 472, 206, 299, and 320, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 1082.

16. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 90, 92, 122, 129, 141 , 143, 144, 145, 150, 151 , 151 / 155, 152, 154, 155, 155 / 265, 157, 160, 161 , 164, 165, 168, 180, 181 , 184, 193, 197, 204, 205, 205 / 472, 206, 299, and 320, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 1082.

17. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 129, 129 / 160, 129 / 160 / 164, 129 / 160 / 184, 129 / 164, 129 / 164 / 184, 129 / 205, 146 / 152 / 244, 150 / 155 / 226, 150 / 155 / 244, 152, 154, 154 / 160 / 184, 154 / 164, 154 / 205, 155, 160, 160 / 164, 160 / 164 / 184, 160 / 184, 164, 164 / 204, 164 / 205, 204, 205, 226, 226 / 244, and 244, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 1218.

18. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 129, 129 / 143, 129 / 154 / 164, 129 / 154 / 164 / 205, 129 / 154 / 205, 129 / 160, 129 / 160 / 164, 129 / 160 / 205, 129 / 164 / 205, 143, 143 / 150 / 155 / 226 / 244, 143 / 150 / 155 / 244, 143 / 150 / 226, 143 / 150 / 226 / 244 / 343, 143 / 150 / 226 / 343, 143 / 150 / 244, 143 / 150 / 343, 143 / 152, 143 / 152 / 226, 143 / 152 / 226 / 244, 143 / 152 / 226 / 343, 143 / 152 / 343, 143 / 155, 143 / 155 / 226 / 244, 143 / 155 / 226 / 244 / 343, 143 / 155 / 343, 143 / 226, 143 / 226 / 244 / 343, 143 / 226 / 343, 143 / 244, 143 / 244 / 343, 143 / 343, 146 / 152 / 244, 150 / 155 / 226, 150 / 155 / 244, 150 / 155 / 244 / 343, 150 / 226 / 244 / 343, 152, 152 / 174 / 244 / 343, 152 / 226, 152 / 226 / 343, 152 / 343, 154, 154 / 160 / 184, 154 / 164, 154 / 164 / 204, 154 / 164 / 204 / 205, 154 / 205, 155, 160, 160 / 164, 160 / 164 / 184, 164, 205, 226, 226 / 244, and 244, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 1218.

19. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 98 / 129 / 157 / 168, 129 / 157 / 168, 151 , 151 / 152 / 255, 151 / 152 / 273, 151 / 255, 151 / 343, 157, 157 / 164, 157 / 164 / 168, 157 / 168, 164, 164 / 255, 168, 183, 255, 255 / 343, and 343, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 1580.

20. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 129, 129 / 157, 129 / 157 / 168, 129 / 164, 129 / 164 / 168 / 183, 129 / 164 / 183, 143, 143 / 151 , 143 / 151 / 152, 143 / 151 / 164, 143 / 164, 143 / 164 / 255, 143 / 164 / 343, 143 / 255, 143 / 343, 151 , 151 / 152, 151 / 152 / 164 / 343, 151 / 152 / 343, 151 / 255, 151 / 255 / 343, 151 / 343, 152, 152 / 255, 157, 157 / 164 / 168, 157 / 168, 164, 164 / 168, 164 / 255, 164 / 447, 168, 183, 255, 255 / 343, 298 / 343, and 343, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 1580.21 . The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 69, 69 / 164, 72, 73, 91 , 92 / 164, 95, 96,97, 98, 110, 112, 113, 121 , 121 / 164, 122, 122 / 164, 124, 126, 127, 139, 140, 141 , 144, 144 / 164, 145, 146, 147, 148, 150, 150 / 164, 151 , 151 / 164, 153, 153 / 164, 155, 156, 157, 159, 160, 161 , 163, 164, 164 / 172, 164 / 173, 164 / 182, 164 / 188, 164 / 191 , 164 / 192, 164 / 195, 164 / 197, 164 / 199, 164 / 202, 164 / 205 / 249, 164 / 209, 165, 166, 167, 169, 170, 171 , 172, 173, 177, 184, 187, 188, 190, 191 , 192, 193, 194, 195, 197,197 / 266, 198, 199, 200, 202, 203, 204, 205, 208, 209, 210, 321 , 324, 331 , and 349, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 1580.

22. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 69, 69 / 164, 72, 73, 91 , 92 / 164, 95, 97,98, 121 / 164, 126, 127, 139, 140, 142, 144, 144 / 164, 145, 146, 146 / 164, 148, 150, 150 / 164, 151 , 153, 153 / 164, 155, 156, 157, 160, 163, 164 / 173, 164 / 188, 164 / 191 , 164 / 192, 164 / 195, 164 / 199, 164 / 205 / 249, 164 / 209, 165, 166, 167, 169, 171 , 172, 173, 177, 182, 187, 188, 190, 191 , 192, 194, 195, 197 / 266, 198, 199, 200, 202, 203, 204, 205, 208, 209, 210, 324, 331 , and 349, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 1580.

23. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 73, 91 , 93, 95, 97, 98, 110, 112, 122, 122 / 164, 124, 127, 139, 144, 145, 146, 146 / 164, 147, 148, 150, 150 / 164, 151 , 151 / 164, 153, 153 / 164, 155, 156, 157, 159, 160, 161 , 162, 163, 164, 164 / 182, 164 / 188, 164 / 191 , 164 / 192, 164 / 195, 164 / 199, 164 / 205 / 249, 164 / 209, 165, 166, 167, 169, 171 , 172, 177, 184, 187, 188, 190, 191 , 192, 193, 195, 197 / 266, 198, 199, 200, 204, 205, 206, 207, 208, 209, 210, and 324, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 1580.

24. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 69, 73, 91 , 127, 140, 144, 145, 146, 146 / 164, 150, 150 / 164, 151 , 153, 155, 156, 157, 160, 163, 164, 164 / 172, 164 / 188, 164 / 191 , 164 / 192, 164 / 195, 164 / 199, 164 / 209, 165, 167, 169, 171 , 172, 182, 183, 188, 191 , 192, 193, 195, 196, 198, 199, 200, 204, 205, 208, 210, and 324, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 1580.

25. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 38 / 143 / 168 / 220 / 290 / 317 / 423,38 / 152 / 154 / 164 / 168 / 220 / 290 / 317 / 423, 38 / 152 / 168 / 220 / 290 / 317 / 343 / 423, 38 / 152 / 168 / 220 / 290 / 317 / 423, 38 / 154 / 164 / 168 / 220 / 290 / 317 / 423, 38 / 168 / 220 / 290 / 317 / 423, and 38 / 220 / 317, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 972.

26. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 73 / 143 / 169 / 191 / 193, 73 / 156, 73 / 156 / 192 / 435, 73 / 156 / 210, 73 / 163, 73 / 163 / 210, 73 / 192 / 193, 127, 127 / 160 / 172 / 198, 127 / 172, 127 / 198, 146, 146 / 151 / 188, 146 / 324, 151 / 171 / 324, 151 / 324, 156 / 169, 156 / 192, 157 / 171 , 163, 163 / 169, 163 / 169 / 191 / 192 / 193, 165 / 171 / 324, 165 / 188 / 324, 165 / 324, 169, 171 / 188 / 324, 171 / 324, 172, 188, 192, 192 / 193, 198, 210, and 324 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 2368.

27. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 73, 73 / 163 / 191 / 192 / 193, 127 / 160 / 172, 127 / 160 / 172 / 198, 144, 160, 160 / 172, 163 / 191 / 192, and 192 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 2368.

28. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 8, 13, 17, 18, 20 / 146 / 157 / 167, 26, 34 / 38, 38 / 39 / 45, 67, 70, 73, 73 / 150 / 155 / 169, 73 / 150 / 171 / 192, 73 / 155, 73 / 155 / 169 / 199, 73 / 155 / 171 , 73 / 192, 73 / 199, 78, 82, 84, 86, 86 / 384, 89, 90, 91 , 95, 102, 110, 111 , 113, 114, 115, 122 / 127, 126 / 127, 127 / 131 / 165, 127 / 133, 143, 143 / 146, 143 / 146 / 153, 143 / 146 / 157, 143 / 146 / 167, 143 / 146 / 167 / 188, 143 / 146 / 188, 143 / 153 / 167 / 188 / 246, 143 / 156 / 157 / 167 / 195, 143 / 156 / 188, 143 / 188, 143 / 188 / 195, 143 / 195, 144, 145, 145 / 151 , 146, 146 / 152, 146 / 153 / 157, 146 / 153 / 157 / 167, 146 / 153 / 167, 146 / 157, 146 / 157 / 167, 146 / 167, 150, 150 / 155 / 169, 150 / 155 / 169 / 192, 150 / 155 / 192, 150 / 169, 150 / 192, 150 / 199, 151 / 152 / 154, 152 / 154, 152 / 154 / 155 / 160, 153 / 157 / 167, 155 / 171 , 155 / 192, 156 / 270, 160 / 371 , 167 / 188, 168 / 172, 169, 184, 190, 192 / 198, 198, 206, 208, 209, 210, 214 / 220, 215 / 220, 216 / 220, 217 / 220, 219 / 220, 220, 220 / 222, 220 / 225, 220 / 226, 231 , 235, 242, 257, 259, 261 , 262, 267, 268, 269, 270, 272, 274, 275, 276, 277, 278, 282, 283 / 290, 284 / 290, 287 / 290, 288 / 290, 289 / 290, 290, 290 / 291 , 290 / 292, 290 / 293, 290 / 294, 304, 305, 305 / 373, 306, 307, 309, 317, 329, 333, 334, 339, 341 , 344, 345, 346, 352, 353, 355, 358, 366, 368, 370, 372, 373, 375, 376, 378, 379, 380, 381 , 382, 383, 384, 385, 386 / 393, 387 / 393, 389 / 393, 390 / 393,391 / 393, 392 / 393, 393 / 394, 414, 415, 420 / 423, 422 / 423, 423, 423 / 425, 432, 440, 441 , 443, 469, and 472 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 2426.

29. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 8, 18, 25, 29, 30, 67, 73,73 / 150 / 155 / 169, 73 / 150 / 155 / 171 , 73 / 150 / 171 / 192, 73 / 171 , 73 / 192, 73 / 199, 74, 82, 86, 86 / 384, 113, 115, 118, 127 / 128, 143, 150, 150 / 155 / 169, 150 / 155 / 169 / 192, 150 / 155 / 171 , 150 / 155 / 171 / 199, 150 / 155 / 192, 150 / 155 / 199, 150 / 192, 150 / 199, 155 / 192, 168 / 171 / 172, 195 / 198, 197 / 198, 198 / 201 , 198 / 203, 198 / 205, 235, 252, 256, 257, 262, 263, 267 / 343, 268, 269, 270, 272, 273, 276, 277, 288 / 290, 289 / 290, 290, 290 / 291 , 290 / 292, 290 / 294, 298, 304, 305, 307, 333, 346, 355, 366, 370, 386 / 393, 389 / 393, 393 / 395, 440, and 472 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 2426.

30. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 66, 67, 70, 101 , 102, 103, 104, 106,107, 108, 113, 114, 115, 117, 118, 143, 152, 154, 235, 236 / 251 , 237, 238, 242, 243, 246, 247, 249, 250, 251 , 256 / 352, 257, 259, 285, 286, 287, 288, 289, 290, 292, 293, 294, 295, 296, 322, 323, 326, 329, 330, 333, 339, 340, 343, 344, 345, and 352. and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 2540.31 . The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 18, 18 / 113 / 270, 18 / 113 / 270 / 329 / 370, 18 / 113 / 270 / 370 / 384, 18 / 113 / 270 / 370 / 412, 18 / 113 / 329 / 384, 18 / 259 / 370 / 384, 18 / 270, 18 / 270 / 370,18 / 270 / 370 / 384, 18 / 370, 18 / 370 / 384, 73, 73 / 115, 73 / 115 / 184 / 333, 73 / 115 / 184 / 341 , 73 / 115 / 256, 73 / 184 / 256, 73 / 184 / 273 / 333, 73 / 184 / 333, 73 / 256, 73 / 256 / 375, 73 / 273, 73 / 273 / 333 / 375, 73 / 273 / 375, 73 / 333, 73 / 333 / 375, 73 / 375, 73 / 379, 77 / 233 / 238 / 376, 77 / 233 / 334 / 376, 77 / 262 / 376, 77 / 276, 77 / 334 / 376, 82, 82 / 127, 82 / 127 / 144 / 235 / 290, 82 / 127 / 144 / 266, 82 / 127 / 144 / 266 / 290, 82 / 127 / 144 / 378, 82 / 127 / 235, 82 / 127 / 290, 82 / 144 / 266 / 290 / 378, 82 / 144 / 266 / 378, 82 / 235 / 266, 82 / 235 / 266 / 378 / 383, 82 / 235 / 290, 82 / 235 / 378, 82 / 266, 82 / 266 / 378, 82 / 378, 113 / 270 / 370 / 384, 115 / 256 / 333 / 375, 115 / 256 / 375, 115 / 273 / 375, 115 / 375, 127 / 144 / 290, 127 / 149 / 150, 127 / 266 / 290 / 378, 127 / 378 / 383, 144 / 235 / 290, 144 / 266 / 290, 233 / 334, 233 / 376, 235, 235 / 290, 256 / 333, 262, 266 / 378, 270 / 370, 273, 276 / 352, 290, 290 / 378, 333, 358, 370, 375, and 378 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 2540.

32. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 18, 18 / 113, 18 / 113 / 252, 18 / 113 / 252 / 336 / 384, 18 / 113 / 252 / 370, 18 / 113 / 270, 18 / 113 / 270 / 370 / 384, 18 / 259 / 370 / 384, 18 / 270,18 / 270 / 370, 18 / 270 / 370 / 384, 18 / 370, 18 / 370 / 384, 73, 73 / 115 / 184 / 341 , 73 / 115 / 341 , 73 / 131 / 256 / 273 / 333 / 341 / 375, 73 / 184 / 256, 73 / 184 / 256 / 341 , 73 / 184 / 256 / 375, 73 / 184 / 273 / 333, 73 / 184 / 333, 73 / 184 / 341 / 375, 73 / 256, 73 / 256 / 273, 73 / 256 / 273 / 375, 73 / 256 / 333 / 375, 73 / 256 / 375, 73 / 273, 73 / 273 / 333 / 341 , 73 / 273 / 333 / 375, 73 / 273 / 375, 73 / 333, 73 / 333 / 341 / 375, 73 / 333 / 375, 73 / 341 , 73 / 375, 73 / 379, 77, 77 / 118, 77 / 118 / 143 / 262 / 276 / 334 / 376, 77 / 118 / 143 / 334 / 352 / 376, 77 / 118 / 143 / 376, 77 / 118 / 233, 77 / 118 / 233 / 352 / 376, 77 / 118 / 262 / 334 / 376, 77 / 118 / 334, 77 / 143 / 262 / 276, 77 / 143 / 262 / 334, 77 / 143 / 376, 77 / 233 / 238 / 376, 77 / 233 / 262, 77 / 233 / 276 / 376, 77 / 233 / 334 / 376, 77 / 262 / 376, 77 / 276, 77 / 334 / 376, 77 / 352, 82, 82 / 127, 82 / 127 / 144 / 235 / 290, 82 / 127 / 144 / 266, 82 / 127 / 144 / 266 / 290, 82 / 127 / 144 / 378, 82 / 127 / 235, 82 / 127 / 235 / 266 / 358, 82 / 127 / 290, 82 / 127 / 290 / 358, 82 / 127 / 290 / 378, 82 / 144 / 266 / 290 / 378, 82 / 144 / 266 / 378, 82 / 150 / 235 / 266, 82 / 235 / 266, 82 / 235 / 266 / 290, 82 / 235 / 266 / 378 / 383, 82 / 235 / 290, 82 / 235 / 378, 82 / 266, 82 / 266 / 378, 82 / 290, 82 / 378, 113, 113 / 252, 113 / 252 / 270 / 370, 113 / 270 / 370 / 384, 113 / 336, 118 / 233 / 276 / 334, 118 / 334, 127 / 235 / 266, 127 / 266 / 290 / 378, 127 / 378 / 383, 143, 143 / 276, 143 / 276 / 334, 144 / 235 / 290, 144 / 266 / 290, 144 / 290 / 358, 233 / 334, 233 / 376, 235, 235 / 266 / 290 / 358 / 383, 235 / 358, 252 / 329 / 336 / 370, 252 / 384, 256 / 333, 256 / 375, 262, 262 / 276, 262 / 334, 266 / 290, 266 / 378, 270 / 370, 273, 276 / 352, 290, 290 / 378, 333, 334, 334 / 376, 336 / 370, 341 , 358, 370, 375, and 378 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 2540.

33. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 18,18 / 77 / 118 / 169 / 233 / 238 / 256 / 266 / 270, 18 / 77 / 150 / 256 / 376, 18 / 118, 18 / 118 / 143 / 169 / 238 / 256 / 266 / 333, 18 / 118 / 150 / 169 / 184 / 238 / 256 / 270 / 378, 18 / 118 / 169, 18 / 118 / 233 / 238, 18 / 143, 18 / 143 / 256, 18 / 150 / 169, 18 / 150 / 266, 18 / 169 / 184 / 290 / 333 / 376, 18 / 169 / 233 / 333, 18 / 169 / 392, 18 / 184 / 270, 18 / 233 / 256 / 334, 18 / 256 / 266 / 290 / 378, 77 / 113 / 118 / 256 / 266 / 290, 77 / 118 / 290, 77 / 169 / 270 / 290, 77 / 233, 77 / 233 / 256, 77 / 238, 113 / 118 / 169 / 184 / 256 / 376, 113 / 169, 113 / 233, 118, 118 / 143 / 238, 118 / 233 / 238 / 256 / 378, 118 / 233 / 238 / 270, 143 / 169 / 233 / 270, 143 / 233 / 266 / 290 / 376, 143 / 238 / 270 / 290 / 378, 143 / 256 / 334, 143 / 290, 169, 169 / 256 / 266, 183 / 270 / 333, 233 / 238, 233 / 238 / 376, 233 / 392, 238 / 256, 256 / 290 / 303, 270, 290, 376, and 378 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 3402.

34. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 18, 18 / 77 / 118 / 169 / 266 / 334 / 376, 18 / 77 / 150 / 256 / 376, 18 / 77 / 256 / 290 / 334, 18 / 113 / 238, 18 / 118, 18 / 118 / 143 / 169 / 238 / 256 / 266 / 333,18 / 118 / 143 / 169 / 376, 18 / 118 / 169, 18 / 118 / 233 / 238, 18 / 118 / 266, 18 / 143, 18 / 143 / 169 / 233 / 238 / 358, 18 / 143 / 233 / 238 / 290 / 376, 18 / 143 / 256, 18 / 143 / 290, 18 / 143 / 333 / 334 / 378, 18 / 150 / 169, 18 / 150 / 266, 18 / 169 / 184 / 290 / 333 / 376, 18 / 169 / 233 / 333, 18 / 169 / 238 / 270 / 334 / 376, 18 / 169 / 392, 18 / 233 / 256 / 334, 18 / 233 / 266 / 290 / 376, 18 / 256 / 266 / 290 / 378, 18 / 266 / 270 / 378, 18 / 378, 77 / 113 / 121 / 143 / 233 / 334, 77 / 118 / 169 / 233 / 266 / 270, 77 / 118 / 238 / 376, 77 / 118 / 290, 77 / 169 / 270 / 290, 77 / 233, 77 / 238, 113 / 238 / 376, 118, 118 / 143 / 238, 118 / 143 / 392, 118 / 169 / 184 / 233 / 266 / 270 / 333, 118 / 233 / 238 / 256 / 378,118 / 256 / 290 / 333 / 334 / 376 / 378, 118 / 256 / 334 / 376, 118 / 266 / 290, 143 / 169 / 233 / 270, 143 / 233 / 266 / 290 / 376, 143 / 238 / 270 / 290 / 378, 143 / 256 / 334, 143 / 290, 143 / 364, 150 / 233 / 333, 169, 169 / 256 / 266, 183 / 270 / 333, 233 / 238, 233 / 238 / 376, 233 / 392, 238 / 256, 238 / 266 / 270 / 378, 256 / 266 / 334, 266 / 270 / 376, 266 / 333, and 378 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 3402.

35. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 77 / 156, 77 / 156 / 270, 77 / 184 / 244, 77 / 184 / 333, 77 / 244 / 270, 77 / 270, 143, 143 / 169 / 238 / 257 / 344, 143 / 169 / 255, 143 / 169 / 255 / 257, 143 / 169 / 343, 143 / 255, 143 / 255 / 257, 143 / 255 / 344, 143 / 257, 156, 156 / 270, 169 / 238, 169 / 238 / 255 / 257 / 344, 169 / 257 / 344, 169 / 344, 184, 184 / 244 / 324, 238, 238 / 255 / 344, 244 / 270, 257 / 344, and 270 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 3662.

36. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 77, 77 / 156 / 184 / 244 / 324, 77 / 184 / 270, 143, 143 / 238, 143 / 238 / 255, 143 / 238 / 255 / 343, 143 / 238 / 255 / 392, 143 / 238 / 344, 143 / 255, 143 / 257 / 392,143 / 344 , 169, 169 / 238 / 255 / 344, 169 / 238 / 344, 169 / 255, 169 / 343, 169 / 392, 238 / 344, 255, 255 / 257, 255 / 257 / 392, 257, and 344 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 3662.

37. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 20, 22, 26, 42, 43, 44, 45, 50, 51 , 52, 53, 54, 55, 60, 62, 64, 65, 95, 97, 99, 103, 104, 111 , 112, 115, 115 / 143 / 169, 115 / 169 / 343, 115 / 343, 116, 116 / 343, 143, 143 / 169, 154 / 439, 169, 169 / 324, 169 / 324 / 343, 169 / 343, 186, 234, 236, 238, 238 / 246, 239, 239 / 244, 241 , 242, 243, 244, 250, 251 , 254, 255, 260, 283, 284, 295, 296, 298 / 326, 319, 322, 323, 324, 324 / 343, 326, 328, 329, 333, 334, 338, 340, 341 , 343, 344, 347, 366, 434, 439, 442, 443, 444, 445, 446, and 447 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 3788.

38. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 26, 26 / 236, 26 / 236 / 260 / 333, 26 / 236 / 295, 26 / 260, 26 / 260 / 295, 26 / 260 / 295 / 333, 26 / 295 / 319, 26 / 333, 64, 64 / 244, 64 / 251 / 260 / 347, 64 / 260 / 295 / 333, 64 / 260 / 347, 103 / 115 / 255, 103 / 115 / 343, 103 / 115 / 343 / 344, 103 / 143 / 255 / 284 / 343 / 344, 103 / 169 / 343 / 344, 103 / 255 / 284, 103 / 343 / 344, 115, 115 / 143 / 255, 115 / 143 / 255 / 343, 115 / 143 / 284,115 / 143 / 343, 115 / 143 / 344, 115 / 169, 115 / 169 / 343, 115 / 255, 115 / 255 / 344, 115 / 284, 115 / 284 / 343 / 344, 115 / 284 / 344, 115 / 343, 115 / 343 / 344, 115 / 344, 143, 143 / 169 / 255 / 284 / 343 / 344, 143 / 169 / 343 / 344, 143 / 255,143 / 255 / 284 / 343 / 344, 143 / 255 / 343, 143 / 255 / 343 / 344, 143 / 255 / 344, 143 / 343 / 344, 169, 169 / 255, 169 / 255 / 284, 169 / 255 / 343, 169 / 284, 169 / 343, 169 / 344, 236, 236 / 244 / 333, 236 / 260, 236 / 347, 244, 244 / 251 , 244 / 251 / 319, 244 / 260, 244 / 295, 244 / 347, 251 , 251 / 260 / 295 / 347, 251 / 333, 255, 255 / 284, 255 / 343, 255 / 343 / 344, 255 / 344, 259, 260, 260 / 295, 260 / 333, 284, 284 / 343 / 344, 284 / 344, 295, 295 / 333, 333, 343, 343 / 344, 344, and 347 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 4150.

39. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 115, 115 / 117, 115 / 117 / 118,115 / 117 / 118 / 152, 115 / 117 / 118 / 169, 115 / 117 / 118 / 246, 115 / 117 / 152, 115 / 117 / 152 / 169, 115 / 117 / 152 / 169 / 246, 115 / 117 / 152 / 246, 115 / 117 / 169, 115 / 117 / 246, 115 / 118, 115 / 118 / 152, 115 / 118 / 152 / 169, 115 / 118 / 152 / 169 / 246, 115 / 118 / 169, 115 / 118 / 246, 115 / 152, 115 / 152 / 169, 115 / 152 / 169 / 246, 115 / 152 / 246, 115 / 152 / 246 / 374, 115 / 169, 115 / 169 / 246, 115 / 246, 117, 117 / 118, 117 / 118 / 152, 117 / 118 / 152 / 169, 117 / 118 / 152 / 246, 117 / 118 / 169, 117 / 118 / 246, 117 / 152, 117 / 152 / 169, 117 / 152 / 246 / 288, 117 / 169, 117 / 246, 118 / 246, 152, 152 / 169, 152 / 169 / 246, 152 / 246, 169, 169 / 246, and 246 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 4278.

40. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 73, 101 , 114, 115, 118, 143, 145, 150, 151 , 152, 153, 154, 156, 157, 160, 161 , 163, 166, 167, 168, 169, 171 , 172, 190, 191 , 192, 195, 213, 214, 218, 219, 223, 225, 233, 233 / 343, 234, 235, 236, 237, 238, 239, 239 / 250, 243, 244, 249, 251 , 254, 255, 257, 258, 259, 261 , 262, 262 / 283, 277 / 292, 283, 284, 286, 289, 292, 295, and 296 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 4278.41 . The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 93, 113, 113 / 114, 113 / 115, 113 / 143, 115 / 118, 115 / 177, 118 / 143 / 163 / 168 / 195 / 255 / 315, 118 / 143 / 255, 118 / 163 / 168 / 214 / 255 / 315, 118 / 168 / 214 / 315, 118 / 255, 124, 126, 129, 134, 143 / 154 / 255, 143 / 156 / 168 / 216 / 255, 143 / 163, 143 / 163 / 255, 143 / 168, 143 / 168 / 169 / 255 / 315, 143 / 255, 148, 156 / 214 / 315, 159, 163, 163 / 168, 163 / 168 / 169, 163 / 168 / 169 / 255 / 315, 163 / 168 / 169 / 315, 163 / 168 / 195 / 214 / 255 / 315, 163 / 214, 163 / 255, 163 / 255 / 315, 168 / 315, 186, 194, 197, 206, 207, 208, 210, 213, 255 / 315, 277, 293, 294, 315, 317, 320, 322, 328, 329, 330, 333, 352, 366, 392, and 445 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 4642.

42. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 127, 128, 129, 140, 180, 182, 194, 277, 293, 294, 299, 300, 317, 319, 320, 324, 333, 356, 366, and 392 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 4642.

43. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 1 13, 1 17, 1 18, 151 , 191 , 196, and 205 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 4984.

44. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 1 14 / 1 15 / 143, 1 14 / 1 15 / 143 / 233, 1 14 / 1 15 / 233, 1 14 / 1 15 / 343, 1 14 / 143, 1 14 / 143 / 151 / 233 / 238 / 261 , 1 14 / 143 / 213 / 261 / 343, 1 14 / 143 / 233, 1 14 / 143 / 233 / 238, 1 14 / 151 / 261 / 343, 1 14 / 213 / 238, 1 14 / 343, 1 15, 1 15 / 143, 1 15 / 143 / 151 / 154 / 213 / 261 ,1 15 / 143 / 151 / 154 / 233 / 343, 1 15 / 143 / 151 / 233 / 261 , 1 15 / 143 / 151 / 233 / 343, 1 15 / 143 / 154 / 213 / 233 / 238 / 261 , 1 15 / 143 / 213, 1 15 / 143 / 233, 1 15 / 143 / 343, 1 15 / 151 / 154 / 233, 1 15 / 151 / 261 , 1 15 / 213 / 233, 1 15 / 213 / 233 / 238, 1 15 / 213 / 261 , 1 15 / 233 / 238, 1 15 / 238, 1 15 / 343, 1 15 / 348, 143, 143 / 151 / 154, 143 / 151 / 154 / 213, 143 / 154, 143 / 213 / 233, 143 / 213 / 233 / 238 / 261 / 343, 143 / 213 / 261 / 343, 143 / 233 / 261 , 143 / 261 , 151 / 154, 151 / 154 / 213 / 343, 151 / 154 / 343, 151 / 213, 151 / 213 / 233, 213 / 233 / 343, and 233 / 238 / 343 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 4984.

45. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 1 17, 1 17 / 1 18 / 143 / 168 / 169 / 392, 1 17 / 1 18 / 143 / 392, 1 17 / 1 18 / 168 / 169, 1 17 / 1 18 / 168 / 169 / 392, 1 17 / 143 / 168 / 169 / 170 / 392,1 17 / 143 / 168 / 169 / 392, 1 17 / 168 / 169, 1 17 / 168 / 169 / 392, 1 17 / 168 / 392, 1 17 / 169, 1 17 / 392, 1 18 / 143, 1 18 / 143 / 168 / 169, 1 18 / 143 / 169, 1 18 / 143 / 392, 1 18 / 168, 1 18 / 168 / 392, 1 18 / 169 / 392, 1 18 / 392, 143 / 168 / 169 / 392, 143 / 169, 143 / 392, 168 / 169, 168 / 169 / 392, 168 / 392, 169 / 392, and 392 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 5196.

46. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 134, 134 / 182 / 194 / 294, 134 / 182 / 277 / 293, 134 / 182 / 293 / 320, 134 / 182 / 294, 134 / 194 / 277, 134 / 194 / 277 / 317, 134 / 194 / 293, 134 / 194 / 293 / 294, 134 / 194 / 293 / 317, 134 / 194 / 294, 134 / 194 / 317, 134 / 194 / 317 / 320, 134 / 317, 182, 182 / 194, 182 / 194 / 293 / 320, 182 / 293 / 294, 194, 194 / 277 / 293 / 317, 194 / 277 / 317 / 320, 194 / 277 / 320, 194 / 293 / 294 / 317,194 / 293 / 320, 277, 293, 317, 317 / 320, and 320 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 5286.

47. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 12 / 134, 13 / 134, 13 / 134 / 210, 22 / 134 / 152, 22 / 134 / 235 / 344, 64 / 93 / 134 / 235, 64 / 134 / 344 / 347, 77 / 134, 79 / 134 / 210, 84 / 134, 86 / 134, 87 / 134, 89 / 134, 93 / 134 / 152 / 235 / 344, 93 / 134 / 235 / 344, 93 / 134 / 244 / 344, 93 / 134 / 344, 131 , 134 / 152 / 344, 134 / 175, 134 / 210 / 382, 134 / 210 / 402, 134 / 210 / 456, 134 / 210 / 478, 134 / 344, 134 / 377, 134 / 378, 134 / 380, 134 / 381 , 134 / 382, 134 / 384, 134 / 386, 134 / 387, 134 / 388, 134 / 390, 134 / 391 , 134 / 393, 134 / 395, 134 / 398, 134 / 401 , 134 / 402, 134 / 406, 134 / 407, 134 / 421 , 134 / 456, 134 / 457, 134 / 459, 134 / 460, 134 / 464, 134 / 467, 134 / 468, 134 / 470, 134 / 471 , 134 / 473, 134 / 473 / 474, 134 / 474, and 134 / 475 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 5412.

48. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 73 / 127 / 134 / 160 / 172 / 198 / 290,73 / 127 / 134 / 160 / 198 / 333, 73 / 127 / 134 / 160 / 210 / 266, 73 / 127 / 134 / 172 / 192 / 266 / 333, 73 / 127 / 134 / 172 / 198, 73 / 127 / 134 / 210 / 290, 73 / 127 / 134 / 210 / 333, 73 / 127 / 134 / 290 / 333, 73 / 127 / 134 / 333, 73 / 134, 73 / 134 / 160 / 172 / 192 / 290, 73 / 134 / 160 / 172 / 198 / 333, 73 / 134 / 160 / 172 / 290, 73 / 134 / 160 / 266, 73 / 134 / 160 / 333, 73 / 134 / 172, 73 / 134 / 172 / 210 / 333, 73 / 134 / 172 / 266 / 333, 73 / 134 / 198 / 210, 73 / 134 / 198 / 210 / 266, 73 / 134 / 210 / 333, 73 / 134 / 333, 73 / 134 / 366, 127 / 134, 127 / 134 / 160 / 172 / 192 / 333, 127 / 134 / 160 / 172 / 210 / 266 / 290, 127 / 134 / 172 / 198 / 266 / 333, 127 / 134 / 192 / 210, 127 / 134 / 198 / 210 / 290, 127 / 134 / 210 / 266 / 290, 127 / 134 / 210 / 333, 134, 134 / 160 / 172 / 333, 134 / 160 / 198 / 266 / 290, 134 / 172 / 192 / 290 / 333, 134 / 192 / 198, 134 / 198, 134 / 198 / 266 / 333, 134 / 210, 134 / 210 / 290, and 134 / 333 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 5412.

49. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 13, 13 / 178 / 381 , 13 / 178 / 381 / 473, 13 / 178 / 457, 13 / 381 , 13 / 381 / 457, 13 / 402 / 457, 13 / 457, 178, 178 / 381 / 457, 178 / 381 / 457 / 473, 178 / 402 / 457, 178 / 473, 381 , 381 / 393 / 457, 381 / 457 / 473, 381 / 473, and 393 / 457 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 5422.

50. The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from38 / 73 / 1 13 / 1 15 / 1 17 / 1 18 / 127 / 134 / 152 / 154 / 160 / 163 / 164 / 168 / 169 / 172 / 184 / 192 / 194 / 198 / 220 / 243 / 260 / 266 / 27 0 / 295 / 317 / 333 / 392 / 423, 38 / 73 / 1 13 / 1 15 / 1 17 / 1 18 / 127 / 134 / 152 / 154 / 160 / 163 / 164 / 168 / 169 / 172 / 184 / 192 / 198 / 220 / 243 / 260 / 266 / 270 / 295 / 317 / 333 / 392 / 423,38 / 73 / 1 13 / 1 15 / 1 17 / 1 18 / 127 / 134 / 152 / 154 / 160 / 163 / 164 / 168 / 169 / 172 / 184 / 192 / 198 / 220 / 243 / 260 / 266 / 270 / 29 5 / 317 / 333 / 392 / 423 / 457,38 / 73 / 1 13 / 1 15 / 1 17 / 1 18 / 127 / 152 / 154 / 160 / 163 / 164 / 168 / 169 / 172 / 184 / 192 / 198 / 220 / 243 / 260 / 266 / 270 / 295 / 31 7 / 333 / 392 / 423,38 / 73 / 1 13 / 1 15 / 1 18 / 127 / 152 / 154 / 160 / 162 / 164 / 168 / 172 / 184 / 192 / 198 / 220 / 243 / 260 / 266 / 270 / 295 / 317 / 333 / 42 3,38 / 73 / 1 13 / 1 15 / 1 18 / 127 / 152 / 154 / 160 / 163 / 164 / 168 / 169 / 172 / 184 / 192 / 198 / 220 / 243 / 260 / 266 / 270 / 295 / 317 / 33 3 / 423,38 / 73 / 1 13 / 1 15 / 1 18 / 127 / 152 / 154 / 160 / 164 / 168 / 172 / 184 / 189 / 192 / 198 / 220 / 243 / 260 / 266 / 270 / 295 / 317 / 333 / 42 3,38 / 73 / 1 13 / 1 15 / 1 18 / 127 / 152 / 154 / 160 / 164 / 168 / 172 / 184 / 192 / 198 / 210 / 220 / 243 / 260 / 266 / 270 / 295 / 317 / 333 / 42 3, 38 / 73 / 1 13 / 1 15 / 1 18 / 127 / 152 / 154 / 160 / 164 / 168 / 172 / 184 / 192 / 198 / 220 / 243 / 260 / 266 / 270 / 295 / 317 / 333 / 423, 38 / 73 / 1 13 / 1 18 / 127 / 152 / 154 / 160 / 164 / 168 / 172 / 184 / 192 / 198 / 220 / 243 / 260 / 266 / 270 / 295 / 317 / 333 / 423, 38 / 73 / 1 13 / 1 18 / 127 / 152 / 154 / 160 / 164 / 168 / 172 / 184 / 192 / 198 / 220 / 243 / 266 / 270 / 317 / 333 / 423, 38 / 73 / 1 13 / 1 18 / 127 / 152 / 154 / 160 / 164 / 168 / 172 / 184 / 192 / 198 / 220 / 266 / 270 / 317 / 333 / 423, and38 / 1 13 / 127 / 152 / 154 / 160 / 164 / 168 / 172 / 198 / 220 / 290 / 317 / 423, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 972.51 . The engineered transposase of Claim 1 , wherein at least one substitution or substitution set comprises substitutions at amino acid positions selected from 48, 1 16, 1 18, 120, 130, 137, 153, 157,163, 175, 181 , 187, 192, 213, 246, 263, 273, 296, 318, 334, 341 , 394, 412, 424, 454, and 458, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 5704.

52. The engineered transposase of any of Claims 1 -51 , wherein said engineered transposase is capable of binding to adapters or donor DNA to form a transposome, cleaving or fragmenting target DNA, and / or ligating the adapters or donor DNA to the target DNA.

53. The engineered transposase of any of Claims 1 -52, having at least one improved property, as compared to a wild-type or reference transposase.

54. The engineered transposase of Claim 53, wherein said improved property is selected from increased polynucleotide fragmentation or cleavage activity, increased thermostability, increased activity at elevated temperatures, increased soluble expression, decreased inhibition, increased adapter loading or adapter ligation (i.e. tagging), increased binding to a target polynucleotide, and / or decreased sequence insertion bias.

55. The engineered transposase of any of Claims 1 -54, wherein said engineered transposase comprises increased polynucleotide fragmentation or cleavage activity as compared to a wild-type or reference transposase.

56. The engineered transposase of any of Claims 1 -54, wherein said engineered transposase comprises increased thermal stability as compared to a wild-type or reference transposase.

57. The engineered transposase of any of Claims 1 -54, wherein said engineered transposase comprises increased activity at elevated temperatures as compared to a wild-type or reference transposase.

58. The engineered transposase of any of Claims 1 -54, wherein said engineered transposase comprises increased adapter loading or adapter ligation (i.e. tagging) as compared to a wild-type or reference transposase.

59. The engineered transposase of any of Claims 1 -54, wherein said engineered transposase comprises increased binding to a target polynucleotide, as compared to a wild-type or reference transposase.

60. The engineered transposase of any of Claims 1 -54, wherein said engineered transposase comprises decreased sequence insertion bias, as compared to a wild-type or reference transposase.61 . The engineered transposase of any of Claims 1 -60, wherein said engineered transposase comprises a polypeptide sequence selected from the polypeptide sequences between SEQ ID NOs: 2- 2368, 2387-5694, and 5706-5756.

62. The engineered transposase of any of Claims 1 -61 , wherein said transposase is purified.

63. A composition comprising at least one engineered transposase of any of Claims 1 -62.

64. A polynucleotide sequence encoding at least one engineered transposase of any of Claims 1 -61 .

65. A polynucleotide sequence comprising at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to the reference sequence of SEQ ID NOs: 1 , 3, 155, 301 , 337, 605, 971 , 1081 , 1217, 1579, 1597, 1599, 1603, 1719, 2367, 2425, 2539, 3401 , 3661 , 3787, 4149, 4277, 4641 , 4983, 5195, 5285, 5411 , 5421 , and / or 5703, and / or or a functional fragment thereof, wherein said polynucleotide sequence encodes an engineered polypeptide comprising at least one substitution at one or more amino acid positions.

66. The polynucleotide sequence of Claim 65, wherein said polynucleotide sequence encodes at least one engineered transposase comprising a sequence having at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to the reference sequence of SEQ ID NOs: 2, 4, 156, 302, 338, 606, 972, 1082, 1218, 1580, 1598, 1600, 1604, 1720, 2368, 2426, 2540, 3402, 3662, 3788, 4150, 4278, 4642, 4984, 5196, 5286, 5412, 5422, and / or 5704.

67. The polynucleotide sequence of Claim 65, wherein said sequence comprises SEQ ID NOs: 1 , 3, 155, 301 , 337, 605, 971 , 1081 , 1217, 1579, 1597, 1599, 1603, 1719, 2367, 2425, 2539, 3401 , 3661 , 3787, 4149, 4277, 4641 , 4983, 5195, 5285, 5411 , 5421 , and / or 5703.

68. The polynucleotide sequence of Claim 64, wherein said polynucleotide sequence is operably linked to a control sequence.

69. The polynucleotide sequence of Claim 64, wherein said polynucleotide sequence is codon-optimized.

70. An expression vector comprising at least one polynucleotide sequence of any of Claims 64-69.71 . A host cell comprising at least one expression vector of Claim 70.

72. A method of producing an engineered transposase polypeptide in a host cell comprising culturing a host cell of Claim 71 , under suitable culture conditions, such that at least one engineered transposase is produced.

73. The method of Claim 72, further comprising recovering at least one engineered transposase from the culture and / or host cells.

74. The method of Claim 72 or 73, further comprising the step of purifying said at least one engineered transposase.

75. A method for generating a mixture of altered nucleic acids comprising exposing a target nucleic acid to an engineered transposase under conditions and for a time sufficient for the engineered transposase to carry out a transposition event, wherein the engineered transposase comprises a polypeptide sequence comprising at least 85% sequence identity to a reference sequence of SEQ ID NOs: 2, 4, 156, 302, 338, 606, 972, 1082, 1218, 1580, 1598, 1600, 1604, 1720, 2368, 2426, 2540, 3402, 3662, 3788, 4150, 4278, 4642, 4984, 5196, 5286, 5412, 5422, and / or 5704, or a functional fragment thereof, wherein the engineered transposase comprises at least one substitution or substitution set in its polypeptide sequence relative to the reference sequence.

76. The method of Claim 75, wherein the altered nucleic acids are fragmented from the target nucleic acid.

77. A method of preparing a nucleic acid library comprising:(a) contacting a target nucleic acid with an engineered transposase under conditions and for a time sufficient for the engineered transposase to carry out a transposition event, wherein the engineered transposase comprises a polypeptide sequence comprising at least 85% sequence identity to a reference sequence of SEQ ID NOs: 2, 4, 156, 302, 338, 606, 972, 1082, 1218, 1580, 1598, 1600, 1604, 1720, 2368, 2426, 2540, 3402, 3662, 3788, 4150, 4278, 4642, 4984, 5196, 5286, 5412, 5422, and / or 5704, or a functional fragment thereof, wherein the engineered transposase comprises at least one substitution or substitution set in its polypeptide sequence relative to the reference sequences;(b) fragmenting the target nucleic acid into a plurality of nucleic acid fragments;(c) selecting a subset of nucleic acid fragments from the plurality of nucleic acid fragments; and(d) amplifying the subset of nucleic acid fragments to generate the nucleic acid library.

78. A method of sequencing a target nucleic acid comprising:(a) contacting the target nucleic acid with an engineered transposase under conditions and for a time sufficient for the engineered transposase to carry out a transposition event, wherein the engineered transposase comprises a polypeptide sequence comprising at least 85% sequence identity to a reference sequence of SEQ ID NOs: 2, 4, 156, 302, 338, 606, 972, 1082, 1218, 1580, 1598, 1600, 1604, 1720, 2368, 2426, 2540, 3402, 3662, 3788, 4150, 4278, 4642, 4984, 5196, 5286, 5412, 5422, and / or 5704, or a functional fragment thereof, wherein the engineered transposase comprises at least one substitution or substitution set in its polypeptide sequence relative to the reference sequence;(b) fragmenting the target nucleic acid into a plurality of nucleic acid fragments;(c) selecting a subset of nucleic acid fragments from the plurality of nucleic acid fragments;(d) amplifying the subset of nucleic acid fragments to generate a nucleic acid library; and(e) sequencing the nucleic acid library.

79. The method of Claim 77 or 78, further comprising adding a polynucleotide to the 5’ and / or 3’ end of each nucleic acid fragment of the plurality of nucleic acid fragments.

80. The method of any one of Claims 75-78, wherein the engineered transposase comprises a polypeptide sequence comprising at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to the reference sequence of SEQ ID NOs: 2, 4, 156, 302, 338, 606, 972, 1082, 1218, 1580, 1598, 1600, 1604, 1720, 2368, 2426, 2540, 3402, 3662, 3788, 4150, 4278, 4642, 4984, 5196, 5286, 5412, 5422, or 5704.81 . The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 2 and the at least one substitution or substitution set comprises substitutions at amino acid positionsselected from 10, 13, 24, 35, 39, 44, 46, 51 , 54, 55, 62 / 361 , 72, 75, 82, 89, 92, 107, 128, 137, 164, 167, 177 / 359, 191 , 192, 193, 194, 201 , 217, 281 , 289, 291 , 291 / 340, 305, 309, 310, 317, 331 , 337, 340, 346, 352, 353, 357, 358, 359, 362, 371 , 396, 414 / 435, 415, 428, 437, 443, 447, and 448, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 2.

82. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 4 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 4, 5, 6, 8, 10, 10 / 317, 10 / 317 / 358, 10 / 317 / 359, 10 / 346, 10 / 346 / 359, 10 / 358, 10 / 359, 12,12 / 373, 13, 15, 16, 18, 19, 26, 30, 30 / 154, 31 , 34, 35, 37, 38, 68, 70, 73, 74, 93, 106, 118, 126, 134, 135, 143, 146, 154 / 453, 163, 182, 207 / 286, 208, 226, 237, 246, 247, 259, 262, 263, 264, 280, 282, 286, 287, 288, 289, 291 , 292, 293, 298, 305, 309, 310, 313, 317, 317 / 346, 317 / 359, 317 / 445, 320, 346, 351 , 355, 356, 358, 359, 360, 368, 369, 410, 411 , 422, 426, 427, 428, 446, 448, 453, and 454, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 4.

83. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 156 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 18 / 73 / 74 / 259, 18 / 73 / 286 / 289 / 411 , 18 / 74 / 118 / 259 / 286 / 289 / 411 , 18 / 74 / 259,18 / 74 / 259 / 286 / 289, 18 / 74 / 259 / 286 / 411 , 18 / 118 / 259, 18 / 259 / 286 / 289 / 411 , 18 / 259 / 289 / 411 , 18 / 289 / 411 , 73 / 74 / 259 / 286 / 289, 73 / 74 / 259 / 289, 73 / 259 / 286, 74 / 259, 74 / 259 / 289, 74 / 286 / 289 / 411 , 259, 259 / 286 / 289, and 289, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 156.

84. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 302 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 16 / 19 / 355 / 427, 16 / 37 / 146 / 286 / 287 / 289 / 310 / 313 / 355, 16 / 146 / 226 / 287 / 289 / 355, 19 / 37 / 146 / 286 / 287 / 310 / 355, 19 / 135 / 286 / 287 / 289 / 355, 19 / 182 / 355, 37 / 70 / 226 / 310 / 313 / 355, 37 / 74 / 310 / 355 / 427, 37 / 135 / 146 / 182 / 226 / 286 / 287 / 355 / 368, 37 / 146 / 226 / 289 / 427, 37 / 146 / 287 / 289 / 427, 38, 41 , 42, 43, 44, 45, 47, 48, 50, 53, 54, 55, 70 / 74 / 226 / 286 / 287 / 289 / 368 / 427, 70 / 74 / 355 / 368, 70 / 135 / 146 / 150 / 182 / 286 / 289 / 313 / 427, 74 / 310 / 313 / 355, 74 / 310 / 355, 74 / 355 / 427, 101 , 103, 117, 135 / 226 / 286 / 289 / 355 / 368, 135 / 355, 146, 146 / 150 / 287 / 289 / 310 / 355, 146 / 182 / 355, 146 / 226 / 286 / 355, 146 / 286 / 287 / 289 / 355 / 427, 146 / 289, 146 / 427, 150 / 226 / 310 / 313, 182 / 226 / 355, 182 / 355, 244, 247, 249, 251 , 287 / 355 / 427, 300, 317 / 411 , 355, 393, 397, 398, 399, 401 , 405, 407, 408, 409, 411 , 413, 426, 429, 431 , 439, 444, 446, 450, 453, 454, 457, 459, 460, 464, 466, 467, 469, 470, 471 , 472, 474, 475, 476, and 478, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 302.

85. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 338 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 16, 16 / 313, 16 / 409 / 411 / 427, 37 / 150 / 408, 37 / 313 / 317 / 408 / 411 , 37 / 317 / 405 / 408, 37 / 317 / 405 / 426, 37 / 405 / 409 / 427, 150, 150 / 313 / 405 / 411 , 150 / 405 / 406, 150 / 408, 150 / 408 / 411 , 226 / 405,317 / 460, and 408 / 426, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 338.

86. The method of any one of claims 75-80, wherein the reference sequence is SEQ ID NO.606 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 12, 13, 18, 19, 22, 26, 31 / 262, 34, 60, 79, 80, 81 , 82, 92, 106, 107, 113, 115, 132, 134, 136,140, 143, 152, 160, 163, 167, 168, 175, 179, 183, 195, 199, 201 , 202, 215, 216, 218, 220, 223, 226, 233,235, 236, 237, 243, 255, 259, 261 , 262, 266, 268, 270, 273, 282, 290, 292, 293, 309, 319, 320, 326, 343,346, 365, 370, 374, 380, 382, 386, 389, 390, 408, 412, 414, 415, 424, and 442, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 606.

87. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO.606 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 2, 12, 13, 15, 16, 18, 19, 22, 26, 30, 31 / 262, 34, 51 , 60, 69, 79, 82, 84, 87, 88, 106, 107, 113, 114, 115, 128, 132, 134, 135, 136, 140, 143, 146, 150, 152, 160, 163, 167, 168, 175, 179, 183, 195,198, 199, 201 , 215, 217, 218, 219, 220, 223, 224, 226, 227, 231 , 233, 235, 236, 243, 254, 255, 259, 261 ,266, 267, 268, 270, 272, 273, 275, 277, 279, 282, 287, 289, 290, 291 , 292, 293, 304, 307, 309, 319, 320,343, 344, 346, 355, 365, 370, 373, 380, 382, 386, 389, 390, 408, 412, 414, 415, 418, 423, 424, and 468, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 606.

88. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 606 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 44 / 45 / 47 / 408 / 453, 44 / 45 / 47 / 453, 44 / 45 / 47 / 453 / 454, 44 / 45 / 103 / 453 / 457, 44 / 45 / 453 / 454 / 457, 44 / 47 / 103 / 393 / 408 / 453 / 454, 44 / 47 / 103 / 453 / 454 / 457, 44 / 47 / 393 / 408 / 453 / 457, 44 / 47 / 393 / 453 / 457,44 / 103 / 408 / 453 / 454, 45 / 47 / 103 / 393 / 453 / 454 / 457, 45 / 47 / 393 / 453 / 454, 45 / 47 / 408 / 423 / 453 / 457, 45 / 47 / 453, 45 / 103 / 393 / 453 / 454, 45 / 453 / 454, 47 / 103 / 408 / 453 / 454, 47 / 393 / 408 / 453 / 454 / 457, 47 / 393 / 453 / 454, 47 / 408 / 453 / 457, 53 / 54 / 244 / 450, 103 / 393 / 408 / 453 / 457, 244 / 450, 408 / 453 / 454, 446 / 450 / 464, and 453 / 457, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 606.

89. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 972 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 38 / 45 / 53, 38 / 45 / 53 / 54 / 476, 38 / 45 / 53 / 244, 38 / 45 / 53 / 470 / 472, 38 / 45 / 54 / 244, 38 / 45 / 54 / 470 / 474 / 478, 38 / 45 / 472 / 476 / 478, 38 / 45 / 476, 38 / 53 / 54, 38 / 53 / 54 / 244, 38 / 53 / 54 / 244 / 472 / 478, 38 / 53 / 472, 38 / 54 / 244 / 470 / 474 / 476 / 478, 38 / 244 / 470 / 472 / 474 / 478, 38 / 478, 45 / 53 / 54 / 244 / 470 / 474 / 476 / 478, 45 / 53 / 54 / 470 / 474 / 476, 45 / 53 / 54 / 472 / 478, 45 / 54 / 244 / 474 / 478, 53, 53 / 54 / 470 / 476 / 478, 53 / 244 / 474 / 478, 54, 54 / 244, 54 / 244 / 472 / 476, 54 / 472, 244, and 474 / 478, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 972.

90. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 972 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 22 / 38 / 45 / 107 / 259 / 317 / 412, 22 / 38 / 45 / 259 / 317 / 412, 22 / 107, 22 / 107 / 317 / 412, 22 / 134, 22 / 134 / 220, 22 / 134 / 220 / 317, 22 / 134 / 317 / 412, 22 / 317, 38, 38 / 45 / 81 / 143 / 244, 38 / 53 / 243, 38 / 81 , 38 / 81 / 244, 38 / 107 / 134 / 220, 38 / 132, 38 / 220 / 317, 45 / 81 , 45 / 81 / 243, 45 / 81 / 243 / 343, 45 / 143 / 243 / 270 / 343, 45 / 244 / 270, 53 / 115 / 163 / 243 / 270 / 343, 53 / 143 / 243 / 270 / 343, 53 / 270, 64 / 134 / 220 / 317 / 412, 81 / 132 / 243, 107, 107 / 134 / 317, 115 / 244 / 270, 134 / 259, 143 / 243 / 270, 243, 243 / 270, 244, 244 / 270, 259, 270, and 317, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 972.91 . The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 972 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 22 / 38 / 45 / 107 / 259 / 317 / 412, 22 / 38 / 45 / 259 / 317 / 412, 22 / 38 / 64 / 134 / 220, 22 / 45 / 220 / 259, 22 / 64 / 107 / 259 / 317 / 412, 22 / 64 / 134 / 220, 22 / 107, 22 / 134, 22 / 134 / 317, 22 / 134 / 317 / 412, 22 / 220, 22 / 220 / 412 / 415, 22 / 259 / 317, 22 / 259 / 317 / 412, 22 / 317, 38, 38 / 45 / 81 / 143 / 244, 38 / 45 / 143 / 270, 38 / 53 / 243, 38 / 81 , 38 / 81 / 143 / 243 / 270, 38 / 81 / 183 / 243 / 270, 38 / 81 / 243, 38 / 81 / 244, 38 / 107 / 134 / 220, 38 / 132, 38 / 220 / 317, 38 / 270 / 343, 45 / 81 , 45 / 81 / 132 / 243 / 270, 45 / 81 / 243, 45 / 143 / 243 / 270 / 343, 45 / 244 / 270, 53 / 143 / 243 / 270 / 343, 53 / 244 / 270, 53 / 270, 64, 64 / 107 / 134, 64 / 107 / 317, 64 / 134, 64 / 134 / 220 / 317 / 412, 81 / 132, 81 / 132 / 243, 107, 107 / 134 / 317, 107 / 412, 115 / 244 / 270, 143 / 243 / 270, 220 / 259, 243, 243 / 270, 244 / 270, 255, 259, and 270, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 972.

92. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 1082 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 143, 143 / 175 / 272 / 273 / 415, 152 / 168 / 216 / 237, 152 / 168 / 423, 160, 160 / 201 / 272, 160 / 201 / 273 / 415, 160 / 226, 160 / 244 / 272 / 273 / 415, 160 / 415, 168, 168 / 187 / 216 / 255 / 423, 168 / 216 / 237, 168 / 255 / 423, 168 / 290 / 423, 187, 187 / 423, 199 / 201 / 226, 199 / 201 / 415, 201 , 216, 216 / 423, 226 / 244 / 415, 226 / 415, 237, 237 / 290, 244, 255, 272, 290, 290 / 423, 415, and 423, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 1082.

93. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 1082 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 143, 143 / 160 / 226 / 415, 143 / 175 / 272 / 273 / 415, 152 / 168 / 216 / 237, 152 / 168 / 423, 160,160 / 201 / 272, 160 / 201 / 273 / 415, 160 / 226, 160 / 244 / 272 / 273 / 415, 160 / 415, 168, 168 / 187 / 216 / 255 / 423, 168 / 216 / 237, 168 / 216 / 237 / 255 / 343, 168 / 255 / 423, 168 / 290 / 423, 187, 199 / 201 / 226, 199 / 201 / 415, 201 , 216, 216 / 423, 226 / 244 / 415, 226 / 415, 237, 237 / 290, 244, 255, 255 / 343, 272, 290, 290 / 423, 415, and 423, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 1082.

94. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 1082 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 72, 90, 91 , 92, 93, 95, 122, 129, 141 , 142, 143, 144, 145, 150, 151 , 151 / 155, 152, 154, 155, 157, 160, 161 , 161 / 313, 164, 165, 168, 180, 181 , 184, 193, 197, 204, 205, 205 / 472, 206, 299, and 320, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 1082.

95. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 1082 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 90, 92, 122, 129, 141 , 143, 144, 145, 150, 151 , 151 / 155, 152, 154, 155, 155 / 265, 157, 160, 161 , 164, 165, 168, 180, 181 , 184, 193, 197, 204, 205, 205 / 472, 206, 299, and 320, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 1082.

96. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 1218 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 129, 129 / 160, 129 / 160 / 164, 129 / 160 / 184, 129 / 164, 129 / 164 / 184, 129 / 205, 146 / 152 / 244, 150 / 155 / 226, 150 / 155 / 244, 152, 154, 154 / 160 / 184, 154 / 164, 154 / 205, 155, 160, 160 / 164, 160 / 164 / 184, 160 / 184, 164, 164 / 204, 164 / 205, 204, 205, 226, 226 / 244, and 244, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 1218.

97. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 1218 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 129, 129 / 143, 129 / 154 / 164, 129 / 154 / 164 / 205, 129 / 154 / 205, 129 / 160, 129 / 160 / 164, 129 / 160 / 205, 129 / 164 / 205, 143, 143 / 150 / 155 / 226 / 244, 143 / 150 / 155 / 244, 143 / 150 / 226, 143 / 150 / 226 / 244 / 343, 143 / 150 / 226 / 343, 143 / 150 / 244, 143 / 150 / 343, 143 / 152, 143 / 152 / 226, 143 / 152 / 226 / 244, 143 / 152 / 226 / 343, 143 / 152 / 343, 143 / 155, 143 / 155 / 226 / 244, 143 / 155 / 226 / 244 / 343, 143 / 155 / 343, 143 / 226, 143 / 226 / 244 / 343, 143 / 226 / 343, 143 / 244, 143 / 244 / 343, 143 / 343, 146 / 152 / 244, 150 / 155 / 226, 150 / 155 / 244, 150 / 155 / 244 / 343, 150 / 226 / 244 / 343, 152, 152 / 174 / 244 / 343, 152 / 226, 152 / 226 / 343, 152 / 343, 154, 154 / 160 / 184, 154 / 164, 154 / 164 / 204, 154 / 164 / 204 / 205, 154 / 205, 155, 160,160 / 164, 160 / 164 / 184, 164, 205, 226, 226 / 244, and 244, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 1218.

98. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 1580 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 98 / 129 / 157 / 168, 129 / 157 / 168, 151 , 151 / 152 / 255, 151 / 152 / 273, 151 / 255, 151 / 343, 157, 157 / 164, 157 / 164 / 168, 157 / 168, 164, 164 / 255, 168, 183, 255, 255 / 343, and 343, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 1580.

99. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 1580 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 129, 129 / 157, 129 / 157 / 168, 129 / 164, 129 / 164 / 168 / 183, 129 / 164 / 183, 143, 143 / 151 , 143 / 151 / 152, 143 / 151 / 164, 143 / 164, 143 / 164 / 255, 143 / 164 / 343, 143 / 255, 143 / 343, 151 , 151 / 152, 151 / 152 / 164 / 343, 151 / 152 / 343, 151 / 255, 151 / 255 / 343, 151 / 343, 152, 152 / 255, 157, 157 / 164 / 168, 157 / 168, 164, 164 / 168, 164 / 255, 164 / 447, 168, 183, 255, 255 / 343, 298 / 343, and 343, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 1580.

100. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 1580 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 69, 69 / 164, 72, 73, 91 , 92 / 164, 95, 96, 97, 98, 110, 112, 113, 121 , 121 / 164, 122, 122 / 164, 124, 126, 127, 139, 140, 141 , 144, 144 / 164, 145, 146, 147, 148, 150, 150 / 164, 151 , 151 / 164, 153, 153 / 164, 155, 156, 157, 159, 160, 161 , 163, 164, 164 / 172, 164 / 173, 164 / 182, 164 / 188, 164 / 191 , 164 / 192, 164 / 195, 164 / 197, 164 / 199, 164 / 202, 164 / 205 / 249, 164 / 209, 165, 166, 167, 169, 170, 171 , 172, 173, 177, 184, 187, 188, 190, 191 , 192, 193, 194, 195, 197, 197 / 266, 198, 199, 200, 202, 203, 204, 205, 208, 209, 210, 321 , 324, 331 , and 349, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 1580.101 . The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 1580 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 69, 69 / 164, 72, 73, 91 , 92 / 164, 95, 97, 98, 121 / 164, 126, 127, 139, 140, 142, 144, 144 / 164, 145, 146, 146 / 164, 148, 150, 150 / 164, 151 , 153, 153 / 164, 155, 156, 157, 160, 163, 164 / 173, 164 / 188, 164 / 191 , 164 / 192, 164 / 195, 164 / 199, 164 / 205 / 249, 164 / 209, 165, 166, 167, 169, 171 , 172, 173, 177, 182, 187, 188, 190, 191 , 192, 194, 195, 197 / 266, 198, 199, 200, 202, 203, 204, 205, 208, 209, 210, 324, 331 , and 349, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 1580.

102. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 1580 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 73, 91 , 93, 95, 97, 98, 110, 112, 122, 122 / 164, 124, 127, 139, 144, 145, 146, 146 / 164, 147, 148, 150, 150 / 164, 151 , 151 / 164, 153, 153 / 164, 155, 156, 157, 159, 160, 161 , 162, 163, 164, 164 / 182, 164 / 188, 164 / 191 , 164 / 192, 164 / 195, 164 / 199, 164 / 205 / 249, 164 / 209, 165, 166, 167, 169, 171 , 172, 177, 184, 187, 188, 190, 191 , 192, 193, 195, 197 / 266, 198, 199, 200, 204, 205, 206, 207, 208, 209, 210, and 324, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 1580.

103. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 1580 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 69, 73, 91 , 127, 140, 144, 145, 146, 146 / 164, 150, 150 / 164, 151 , 153, 155, 156, 157, 160,163, 164, 164 / 172, 164 / 188, 164 / 191 , 164 / 192, 164 / 195, 164 / 199, 164 / 209, 165, 167, 169, 171 , 172, 182, 183, 188, 191 , 192, 193, 195, 196, 198, 199, 200, 204, 205, 208, 210, and 324, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 1580.

104. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 972 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 38 / 143 / 168 / 220 / 290 / 317 / 423, 38 / 152 / 154 / 164 / 168 / 220 / 290 / 317 / 423,38 / 152 / 168 / 220 / 290 / 317 / 343 / 423, 38 / 152 / 168 / 220 / 290 / 317 / 423, 38 / 154 / 164 / 168 / 220 / 290 / 317 / 423, 38 / 168 / 220 / 290 / 317 / 423, and 38 / 220 / 317, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 972.

105. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 2368 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 73 / 143 / 169 / 191 / 193, 73 / 156, 73 / 156 / 192 / 435, 73 / 156 / 210, 73 / 163, 73 / 163 / 210, 73 / 192 / 193, 127, 127 / 160 / 172 / 198, 127 / 172, 127 / 198, 146, 146 / 151 / 188, 146 / 324, 151 / 171 / 324, 151 / 324, 156 / 169, 156 / 192, 157 / 171 , 163, 163 / 169, 163 / 169 / 191 / 192 / 193, 165 / 171 / 324, 165 / 188 / 324, 165 / 324, 169, 171 / 188 / 324, 171 / 324, 172, 188, 192, 192 / 193, 198, 210, and 324 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 2368.

106. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 2368 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 73, 73 / 163 / 191 / 192 / 193, 127 / 160 / 172, 127 / 160 / 172 / 198, 144, 160, 160 / 172, 163 / 191 / 192, and 192 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 2368.

107. The method of any one of claims 75-80, wherein the reference sequence is SEQ ID NO. 2426 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 8, 13, 17, 18, 20 / 146 / 157 / 167, 26, 34 / 38, 38 / 39 / 45, 67, 70, 73, 73 / 150 / 155 / 169, 73 / 150 / 171 / 192, 73 / 155, 73 / 155 / 169 / 199, 73 / 155 / 171 , 73 / 192, 73 / 199, 78, 82, 84, 86, 86 / 384, 89, 90, 91 , 95, 102, 110, 111 , 113, 114, 115, 122 / 127, 126 / 127, 127 / 131 / 165, 127 / 133, 143, 143 / 146, 143 / 146 / 153, 143 / 146 / 157, 143 / 146 / 167, 143 / 146 / 167 / 188, 143 / 146 / 188, 143 / 153 / 167 / 188 / 246, 143 / 156 / 157 / 167 / 195, 143 / 156 / 188, 143 / 188, 143 / 188 / 195, 143 / 195, 144, 145, 145 / 151 , 146, 146 / 152, 146 / 153 / 157, 146 / 153 / 157 / 167, 146 / 153 / 167, 146 / 157, 146 / 157 / 167, 146 / 167, 150, 150 / 155 / 169, 150 / 155 / 169 / 192, 150 / 155 / 192, 150 / 169, 150 / 192, 150 / 199, 151 / 152 / 154, 152 / 154, 152 / 154 / 155 / 160, 153 / 157 / 167,155 / 171 , 155 / 192, 156 / 270, 160 / 371 , 167 / 188, 168 / 172, 169, 184, 190, 192 / 198, 198, 206, 208, 209, 210, 214 / 220, 215 / 220, 216 / 220, 217 / 220, 219 / 220, 220, 220 / 222, 220 / 225, 220 / 226, 231 , 235, 242, 257, 259, 261 , 262, 267, 268, 269, 270, 272, 274, 275, 276, 277, 278, 282, 283 / 290, 284 / 290, 287 / 290, 288 / 290, 289 / 290, 290, 290 / 291 , 290 / 292, 290 / 293, 290 / 294, 304, 305, 305 / 373, 306, 307, 309, 317, 329, 333, 334, 339, 341 , 344, 345, 346, 352, 353, 355, 358, 366, 368, 370, 372, 373, 375, 376, 378, 379, 380, 381 , 382, 383, 384, 385, 386 / 393, 387 / 393, 389 / 393, 390 / 393, 391 / 393, 392 / 393, 393 / 394, 414, 415, 420 / 423,422 / 423, 423, 423 / 425, 432, 440, 441 , 443, 469, and 472 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 2426.

108. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 2426 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 8, 18, 25, 29, 30, 67, 73, 73 / 150 / 155 / 169, 73 / 150 / 155 / 171 , 73 / 150 / 171 / 192, 73 / 171 , 73 / 192, 73 / 199, 74, 82, 86, 86 / 384, 113, 115, 118, 127 / 128, 143, 150, 150 / 155 / 169, 150 / 155 / 169 / 192, 150 / 155 / 171 , 150 / 155 / 171 / 199, 150 / 155 / 192, 150 / 155 / 199, 150 / 192, 150 / 199, 155 / 192, 168 / 171 / 172, 195 / 198, 197 / 198, 198 / 201 , 198 / 203, 198 / 205, 235, 252, 256, 257, 262, 263, 267 / 343, 268, 269, 270, 272, 273, 276, 277, 288 / 290, 289 / 290, 290, 290 / 291 , 290 / 292, 290 / 294, 298, 304, 305, 307, 333, 346, 355, 366, 370, 386 / 393, 389 / 393, 393 / 395, 440, and 472 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 2426.

109. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 2540 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 66, 67, 70, 101 , 102, 103, 104, 106, 107, 108, 113, 114, 115, 117, 118, 143, 152, 154, 235, 236 / 251 , 237, 238, 242, 243, 246, 247, 249, 250, 251 , 256 / 352, 257, 259, 285, 286, 287, 288, 289, 290, 292, 293, 294, 295, 296, 322, 323, 326, 329, 330, 333, 339, 340, 343, 344, 345, and 352. and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 2540.

110. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 2540 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 18, 18 / 113 / 270, 18 / 113 / 270 / 329 / 370, 18 / 113 / 270 / 370 / 384, 18 / 113 / 270 / 370 / 412, 18 / 113 / 329 / 384, 18 / 259 / 370 / 384, 18 / 270, 18 / 270 / 370, 18 / 270 / 370 / 384, 18 / 370, 18 / 370 / 384, 73, 73 / 115, 73 / 115 / 184 / 333, 73 / 115 / 184 / 341 , 73 / 115 / 256, 73 / 184 / 256, 73 / 184 / 273 / 333, 73 / 184 / 333, 73 / 256, 73 / 256 / 375, 73 / 273, 73 / 273 / 333 / 375, 73 / 273 / 375, 73 / 333, 73 / 333 / 375, 73 / 375, 73 / 379, 77 / 233 / 238 / 376, 77 / 233 / 334 / 376, 77 / 262 / 376, 77 / 276, 77 / 334 / 376, 82, 82 / 127, 82 / 127 / 144 / 235 / 290, 82 / 127 / 144 / 266, 82 / 127 / 144 / 266 / 290, 82 / 127 / 144 / 378, 82 / 127 / 235, 82 / 127 / 290, 82 / 144 / 266 / 290 / 378, 82 / 144 / 266 / 378, 82 / 235 / 266, 82 / 235 / 266 / 378 / 383, 82 / 235 / 290, 82 / 235 / 378, 82 / 266, 82 / 266 / 378, 82 / 378, 113 / 270 / 370 / 384, 115 / 256 / 333 / 375, 115 / 256 / 375, 115 / 273 / 375, 115 / 375, 127 / 144 / 290, 127 / 149 / 150, 127 / 266 / 290 / 378, 127 / 378 / 383, 144 / 235 / 290, 144 / 266 / 290, 233 / 334, 233 / 376, 235, 235 / 290, 256 / 333, 262, 266 / 378, 270 / 370, 273, 276 / 352, 290, 290 / 378, 333, 358, 370, 375, and 378 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 2540.

111. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 2540 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 18, 18 / 113, 18 / 113 / 252, 18 / 113 / 252 / 336 / 384, 18 / 113 / 252 / 370, 18 / 113 / 270, 18 / 113 / 270 / 370 / 384, 18 / 259 / 370 / 384, 18 / 270, 18 / 270 / 370, 18 / 270 / 370 / 384, 18 / 370, 18 / 370 / 384, 73, 73 / 115 / 184 / 341 , 73 / 115 / 341 , 73 / 131 / 256 / 273 / 333 / 341 / 375, 73 / 184 / 256, 73 / 184 / 256 / 341 , 73 / 184 / 256 / 375,73 / 184 / 273 / 333, 73 / 184 / 333, 73 / 184 / 341 / 375, 73 / 256, 73 / 256 / 273, 73 / 256 / 273 / 375, 73 / 256 / 333 / 375, 73 / 256 / 375, 73 / 273, 73 / 273 / 333 / 341 , 73 / 273 / 333 / 375, 73 / 273 / 375, 73 / 333, 73 / 333 / 341 / 375, 73 / 333 / 375, 73 / 341 , 73 / 375, 73 / 379, 77, 77 / 118, 77 / 118 / 143 / 262 / 276 / 334 / 376, 77 / 118 / 143 / 334 / 352 / 376, 77 / 118 / 143 / 376, 77 / 118 / 233, 77 / 118 / 233 / 352 / 376, 77 / 118 / 262 / 334 / 376, 77 / 118 / 334, 77 / 143 / 262 / 276, 77 / 143 / 262 / 334, 77 / 143 / 376, 77 / 233 / 238 / 376, 77 / 233 / 262, 77 / 233 / 276 / 376, 77 / 233 / 334 / 376, 77 / 262 / 376, 77 / 276, 77 / 334 / 376, 77 / 352, 82, 82 / 127, 82 / 127 / 144 / 235 / 290, 82 / 127 / 144 / 266, 82 / 127 / 144 / 266 / 290, 82 / 127 / 144 / 378, 82 / 127 / 235, 82 / 127 / 235 / 266 / 358, 82 / 127 / 290, 82 / 127 / 290 / 358, 82 / 127 / 290 / 378, 82 / 144 / 266 / 290 / 378, 82 / 144 / 266 / 378, 82 / 150 / 235 / 266, 82 / 235 / 266, 82 / 235 / 266 / 290, 82 / 235 / 266 / 378 / 383, 82 / 235 / 290, 82 / 235 / 378, 82 / 266, 82 / 266 / 378, 82 / 290, 82 / 378, 113, 113 / 252, 113 / 252 / 270 / 370, 113 / 270 / 370 / 384, 113 / 336, 118 / 233 / 276 / 334, 118 / 334, 127 / 235 / 266, 127 / 266 / 290 / 378, 127 / 378 / 383, 143, 143 / 276, 143 / 276 / 334, 144 / 235 / 290, 144 / 266 / 290, 144 / 290 / 358, 233 / 334, 233 / 376, 235, 235 / 266 / 290 / 358 / 383, 235 / 358, 252 / 329 / 336 / 370, 252 / 384, 256 / 333, 256 / 375, 262, 262 / 276, 262 / 334, 266 / 290, 266 / 378, 270 / 370, 273, 276 / 352, 290, 290 / 378, 333, 334, 334 / 376, 336 / 370, 341 , 358, 370, 375, and 378 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 2540.

112. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 3402 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 18, 18 / 77 / 118 / 169 / 233 / 238 / 256 / 266 / 270, 18 / 77 / 150 / 256 / 376, 18 / 118,18 / 118 / 143 / 169 / 238 / 256 / 266 / 333, 18 / 118 / 150 / 169 / 184 / 238 / 256 / 270 / 378, 18 / 118 / 169, 18 / 118 / 233 / 238, 18 / 143, 18 / 143 / 256, 18 / 150 / 169, 18 / 150 / 266, 18 / 169 / 184 / 290 / 333 / 376, 18 / 169 / 233 / 333, 18 / 169 / 392, 18 / 184 / 270, 18 / 233 / 256 / 334, 18 / 256 / 266 / 290 / 378, 77 / 113 / 118 / 256 / 266 / 290, 77 / 118 / 290, 77 / 169 / 270 / 290, 77 / 233, 77 / 233 / 256, 77 / 238, 113 / 118 / 169 / 184 / 256 / 376, 113 / 169, 113 / 233, 118, 118 / 143 / 238, 118 / 233 / 238 / 256 / 378, 118 / 233 / 238 / 270, 143 / 169 / 233 / 270, 143 / 233 / 266 / 290 / 376, 143 / 238 / 270 / 290 / 378, 143 / 256 / 334, 143 / 290, 169, 169 / 256 / 266, 183 / 270 / 333, 233 / 238, 233 / 238 / 376, 233 / 392, 238 / 256, 256 / 290 / 303, 270, 290, 376, and 378 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 3402.

113. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 3402 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 18, 18 / 77 / 118 / 169 / 266 / 334 / 376, 18 / 77 / 150 / 256 / 376, 18 / 77 / 256 / 290 / 334, 18 / 113 / 238,18 / 118, 18 / 118 / 143 / 169 / 238 / 256 / 266 / 333, 18 / 118 / 143 / 169 / 376, 18 / 118 / 169, 18 / 118 / 233 / 238, 18 / 118 / 266, 18 / 143, 18 / 143 / 169 / 233 / 238 / 358, 18 / 143 / 233 / 238 / 290 / 376, 18 / 143 / 256, 18 / 143 / 290, 18 / 143 / 333 / 334 / 378, 18 / 150 / 169, 18 / 150 / 266, 18 / 169 / 184 / 290 / 333 / 376, 18 / 169 / 233 / 333, 18 / 169 / 238 / 270 / 334 / 376, 18 / 169 / 392, 18 / 233 / 256 / 334, 18 / 233 / 266 / 290 / 376, 18 / 256 / 266 / 290 / 378, 18 / 266 / 270 / 378, 18 / 378, 77 / 113 / 121 / 143 / 233 / 334, 77 / 118 / 169 / 233 / 266 / 270, 77 / 118 / 238 / 376, 77 / 118 / 290, 77 / 169 / 270 / 290, 77 / 233, 77 / 238, 113 / 238 / 376, 118, 118 / 143 / 238, 118 / 143 / 392, 118 / 169 / 184 / 233 / 266 / 270 / 333, 118 / 233 / 238 / 256 / 378, 118 / 256 / 290 / 333 / 334 / 376 / 378, 118 / 256 / 334 / 376, 118 / 266 / 290, 143 / 169 / 233 / 270, 143 / 233 / 266 / 290 / 376, 143 / 238 / 270 / 290 / 378, 143 / 256 / 334, 143 / 290, 143 / 364, 150 / 233 / 333, 169, 169 / 256 / 266, 183 / 270 / 333, 233 / 238, 233 / 238 / 376, 233 / 392, 238 / 256, 238 / 266 / 270 / 378, 256 / 266 / 334,266 / 270 / 376, 266 / 333, and 378 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 3402.

114. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 3662 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 77 / 156, 77 / 156 / 270, 77 / 184 / 244, 77 / 184 / 333, 77 / 244 / 270, 77 / 270, 143, 143 / 169 / 238 / 257 / 344, 143 / 169 / 255, 143 / 169 / 255 / 257, 143 / 169 / 343, 143 / 255, 143 / 255 / 257, 143 / 255 / 344, 143 / 257, 156, 156 / 270, 169 / 238, 169 / 238 / 255 / 257 / 344, 169 / 257 / 344, 169 / 344, 184, 184 / 244 / 324, 238, 238 / 255 / 344, 244 / 270, 257 / 344, and 270 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 3662.

115. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 3662 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 77, 77 / 156 / 184 / 244 / 324, 77 / 184 / 270, 143, 143 / 238, 143 / 238 / 255, 143 / 238 / 255 / 343, 143 / 238 / 255 / 392, 143 / 238 / 344, 143 / 255, 143 / 257 / 392, 143 / 344, 169, 169 / 238 / 255 / 344, 169 / 238 / 344, 169 / 255, 169 / 343, 169 / 392, 238 / 344, 255, 255 / 257, 255 / 257 / 392, 257, and 344 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 3662.

116. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 3788 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 20, 22, 26, 42, 43, 44, 45, 50, 51 , 52, 53, 54, 55, 60, 62, 64, 65, 95, 97, 99, 103, 104, 111 , 112, 115, 115 / 143 / 169, 115 / 169 / 343, 115 / 343, 116, 116 / 343, 143, 143 / 169, 154 / 439, 169, 169 / 324, 169 / 324 / 343, 169 / 343, 186, 234, 236, 238, 238 / 246, 239, 239 / 244, 241 , 242, 243, 244, 250, 251 , 254, 255, 260, 283, 284, 295, 296, 298 / 326, 319, 322, 323, 324, 324 / 343, 326, 328, 329, 333, 334, 338, 340, 341 , 343, 344, 347, 366, 434, 439, 442, 443, 444, 445, 446, and 447 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 3788.

117. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 4150 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 26, 26 / 236, 26 / 236 / 260 / 333, 26 / 236 / 295, 26 / 260, 26 / 260 / 295, 26 / 260 / 295 / 333, 26 / 295 / 319, 26 / 333, 64, 64 / 244, 64 / 251 / 260 / 347, 64 / 260 / 295 / 333, 64 / 260 / 347, 103 / 115 / 255, 103 / 115 / 343, 103 / 115 / 343 / 344, 103 / 143 / 255 / 284 / 343 / 344, 103 / 169 / 343 / 344, 103 / 255 / 284, 103 / 343 / 344, 115,115 / 143 / 255, 115 / 143 / 255 / 343, 115 / 143 / 284, 115 / 143 / 343, 115 / 143 / 344, 115 / 169, 115 / 169 / 343, 115 / 255, 115 / 255 / 344, 115 / 284, 115 / 284 / 343 / 344, 115 / 284 / 344, 115 / 343, 115 / 343 / 344, 115 / 344, 143, 143 / 169 / 255 / 284 / 343 / 344, 143 / 169 / 343 / 344, 143 / 255, 143 / 255 / 284 / 343 / 344, 143 / 255 / 343, 143 / 255 / 343 / 344, 143 / 255 / 344, 143 / 343 / 344, 169, 169 / 255, 169 / 255 / 284, 169 / 255 / 343, 169 / 284, 169 / 343, 169 / 344, 236, 236 / 244 / 333, 236 / 260, 236 / 347, 244, 244 / 251 , 244 / 251 / 319, 244 / 260, 244 / 295, 244 / 347, 251 , 251 / 260 / 295 / 347, 251 / 333, 255, 255 / 284, 255 / 343, 255 / 343 / 344, 255 / 344, 259, 260, 260 / 295, 260 / 333, 284, 284 / 343 / 344, 284 / 344, 295, 295 / 333, 333, 343, 343 / 344, 344, and 347 and / or anycombinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 4150.

118. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO.4278 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 115, 115 / 117, 115 / 117 / 118, 115 / 117 / 118 / 152, 115 / 117 / 118 / 169, 115 / 117 / 118 / 246, 115 / 117 / 152, 115 / 117 / 152 / 169, 115 / 117 / 152 / 169 / 246, 115 / 117 / 152 / 246, 115 / 117 / 169, 115 / 117 / 246, 115 / 118, 115 / 118 / 152, 115 / 118 / 152 / 169, 115 / 118 / 152 / 169 / 246, 115 / 118 / 169, 115 / 118 / 246, 115 / 152, 115 / 152 / 169, 115 / 152 / 169 / 246, 115 / 152 / 246, 115 / 152 / 246 / 374, 115 / 169, 115 / 169 / 246, 115 / 246, 117, 117 / 118, 117 / 118 / 152, 117 / 118 / 152 / 169, 117 / 118 / 152 / 246, 117 / 118 / 169, 117 / 118 / 246, 117 / 152, 117 / 152 / 169, 117 / 152 / 246 / 288, 117 / 169, 117 / 246, 118 / 246, 152, 152 / 169, 152 / 169 / 246, 152 / 246, 169, 169 / 246, and 246 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 4278.

119. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 4278 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 73, 101 , 114, 115, 118, 143, 145, 150, 151 , 152, 153, 154, 156, 157, 160, 161 , 163, 166,167, 168, 169, 171 , 172, 190, 191 , 192, 195, 213, 214, 218, 219, 223, 225, 233, 233 / 343, 234, 235, 236, 237, 238, 239, 239 / 250, 243, 244, 249, 251 , 254, 255, 257, 258, 259, 261 , 262, 262 / 283, 277 / 292, 283, 284, 286, 289, 292, 295, and 296 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 4278.

120. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 4642 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 93, 113, 113 / 114, 113 / 115, 113 / 143, 115 / 118, 115 / 177, 118 / 143 / 163 / 168 / 195 / 255 / 315, 118 / 143 / 255, 118 / 163 / 168 / 214 / 255 / 315, 118 / 168 / 214 / 315, 118 / 255, 124, 126, 129, 134, 143 / 154 / 255, 143 / 156 / 168 / 216 / 255, 143 / 163, 143 / 163 / 255, 143 / 168, 143 / 168 / 169 / 255 / 315, 143 / 255, 148, 156 / 214 / 315, 159, 163, 163 / 168, 163 / 168 / 169, 163 / 168 / 169 / 255 / 315, 163 / 168 / 169 / 315, 163 / 168 / 195 / 214 / 255 / 315,163 / 214, 163 / 255, 163 / 255 / 315, 168 / 315, 186, 194, 197, 206, 207, 208, 210, 213, 255 / 315, 277, 293, 294, 315, 317, 320, 322, 328, 329, 330, 333, 352, 366, 392, and 445 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 4642.121 . The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 4642 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 127, 128, 129, 140, 180, 182, 194, 277, 293, 294, 299, 300, 317, 319, 320, 324, 333, 356, 366, and 392 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 4642.

122. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 4984 and the at least one substitution or substitution set comprises substitutions at amino acid positionsselected from 1 13, 1 17, 1 18, 151 , 191 , 196, and 205 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 4984.

123. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 4984 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 1 14 / 1 15 / 143, 1 14 / 1 15 / 143 / 233, 1 14 / 1 15 / 233, 1 14 / 1 15 / 343, 1 14 / 143, 1 14 / 143 / 151 / 233 / 238 / 261 , 1 14 / 143 / 213 / 261 / 343, 1 14 / 143 / 233, 1 14 / 143 / 233 / 238, 1 14 / 151 / 261 / 343, 1 14 / 213 / 238, 1 14 / 343, 1 15, 1 15 / 143, 1 15 / 143 / 151 / 154 / 213 / 261 , 1 15 / 143 / 151 / 154 / 233 / 343,1 15 / 143 / 151 / 233 / 261 , 1 15 / 143 / 151 / 233 / 343, 1 15 / 143 / 154 / 213 / 233 / 238 / 261 , 1 15 / 143 / 213, 1 15 / 143 / 233, 1 15 / 143 / 343, 1 15 / 151 / 154 / 233, 1 15 / 151 / 261 , 1 15 / 213 / 233, 1 15 / 213 / 233 / 238, 1 15 / 213 / 261 , 1 15 / 233 / 238, 1 15 / 238, 1 15 / 343, 1 15 / 348, 143, 143 / 151 / 154, 143 / 151 / 154 / 213, 143 / 154, 143 / 213 / 233, 143 / 213 / 233 / 238 / 261 / 343, 143 / 213 / 261 / 343, 143 / 233 / 261 , 143 / 261 , 151 / 154, 151 / 154 / 213 / 343, 151 / 154 / 343, 151 / 213, 151 / 213 / 233, 213 / 233 / 343, and 233 / 238 / 343 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 4984.

124. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 5196 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 1 17, 1 17 / 1 18 / 143 / 168 / 169 / 392, 1 17 / 1 18 / 143 / 392, 1 17 / 1 18 / 168 / 169, 1 17 / 1 18 / 168 / 169 / 392, 1 17 / 143 / 168 / 169 / 170 / 392, 1 17 / 143 / 168 / 169 / 392, 1 17 / 168 / 169, 1 17 / 168 / 169 / 392, 1 17 / 168 / 392, 1 17 / 169, 1 17 / 392, 1 18 / 143, 1 18 / 143 / 168 / 169, 1 18 / 143 / 169, 1 18 / 143 / 392, 1 18 / 168, 1 18 / 168 / 392, 1 18 / 169 / 392, 1 18 / 392, 143 / 168 / 169 / 392, 143 / 169, 143 / 392, 168 / 169, 168 / 169 / 392, 168 / 392, 169 / 392, and 392 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 5196.

125. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 5286 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 134, 134 / 182 / 194 / 294, 134 / 182 / 277 / 293, 134 / 182 / 293 / 320, 134 / 182 / 294, 134 / 194 / 277, 134 / 194 / 277 / 317, 134 / 194 / 293, 134 / 194 / 293 / 294, 134 / 194 / 293 / 317, 134 / 194 / 294, 134 / 194 / 317, 134 / 194 / 317 / 320, 134 / 317, 182, 182 / 194, 182 / 194 / 293 / 320, 182 / 293 / 294, 194, 194 / 277 / 293 / 317, 194 / 277 / 317 / 320, 194 / 277 / 320, 194 / 293 / 294 / 317, 194 / 293 / 320, 277, 293, 317, 317 / 320, and 320 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 5286.

126. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 5412 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 12 / 134, 13 / 134, 13 / 134 / 210, 22 / 134 / 152, 22 / 134 / 235 / 344, 64 / 93 / 134 / 235, 64 / 134 / 344 / 347, 77 / 134, 79 / 134 / 210, 84 / 134, 86 / 134, 87 / 134, 89 / 134, 93 / 134 / 152 / 235 / 344, 93 / 134 / 235 / 344, 93 / 134 / 244 / 344, 93 / 134 / 344, 131 , 134 / 152 / 344, 134 / 175, 134 / 210 / 382, 134 / 210 / 402, 134 / 210 / 456, 134 / 210 / 478, 134 / 344, 134 / 377, 134 / 378, 134 / 380, 134 / 381 , 134 / 382, 134 / 384, 134 / 386, 134 / 387, 134 / 388, 134 / 390, 134 / 391 , 134 / 393, 134 / 395, 134 / 398, 134 / 401 , 134 / 402, 134 / 406, 134 / 407, 134 / 421 ,134 / 456, 134 / 457, 134 / 459, 134 / 460, 134 / 464, 134 / 467, 134 / 468, 134 / 470, 134 / 471 , 134 / 473, 134 / 473 / 474, 134 / 474, and 134 / 475 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 5412.

127. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 5412 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 73 / 127 / 134 / 160 / 172 / 198 / 290, 73 / 127 / 134 / 160 / 198 / 333, 73 / 127 / 134 / 160 / 210 / 266,73 / 127 / 134 / 172 / 192 / 266 / 333, 73 / 127 / 134 / 172 / 198, 73 / 127 / 134 / 210 / 290, 73 / 127 / 134 / 210 / 333, 73 / 127 / 134 / 290 / 333, 73 / 127 / 134 / 333, 73 / 134, 73 / 134 / 160 / 172 / 192 / 290, 73 / 134 / 160 / 172 / 198 / 333, 73 / 134 / 160 / 172 / 290, 73 / 134 / 160 / 266, 73 / 134 / 160 / 333, 73 / 134 / 172, 73 / 134 / 172 / 210 / 333, 73 / 134 / 172 / 266 / 333, 73 / 134 / 198 / 210, 73 / 134 / 198 / 210 / 266, 73 / 134 / 210 / 333, 73 / 134 / 333, 73 / 134 / 366, 127 / 134, 127 / 134 / 160 / 172 / 192 / 333, 127 / 134 / 160 / 172 / 210 / 266 / 290, 127 / 134 / 172 / 198 / 266 / 333, 127 / 134 / 192 / 210, 127 / 134 / 198 / 210 / 290, 127 / 134 / 210 / 266 / 290, 127 / 134 / 210 / 333, 134, 134 / 160 / 172 / 333, 134 / 160 / 198 / 266 / 290, 134 / 172 / 192 / 290 / 333, 134 / 192 / 198, 134 / 198, 134 / 198 / 266 / 333, 134 / 210, 134 / 210 / 290, and 134 / 333 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 5412.

128. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 5422 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 13, 13 / 178 / 381 , 13 / 178 / 381 / 473, 13 / 178 / 457, 13 / 381 , 13 / 381 / 457, 13 / 402 / 457, 13 / 457, 178, 178 / 381 / 457, 178 / 381 / 457 / 473, 178 / 402 / 457, 178 / 473, 381 , 381 / 393 / 457, 381 / 457 / 473, 381 / 473, and 393 / 457 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 5422.

129. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 972 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from38 / 73 / 113 / 115 / 117 / 118 / 127 / 134 / 152 / 154 / 160 / 163 / 164 / 168 / 169 / 172 / 184 / 192 / 198 / 220 / 243 / 260 / 266 / 270 / 29 5 / 317 / 333 / 392 / 423, 38 / 73 / 113 / 115 / 117 / 118 / 127 / 134 / 152 / 154 / 160 / 163 / 164 / 168 / 169 / 172 / 184 / 192 / 194 / 198 / 220 / 243 / 260 / 266 / 27 0 / 295 / 317 / 333 / 392 / 423,38 / 73 / 113 / 115 / 117 / 118 / 127 / 134 / 152 / 154 / 160 / 163 / 164 / 168 / 169 / 172 / 184 / 192 / 198 / 220 / 243 / 260 / 266 / 270 / 29 5 / 317 / 333 / 392 / 423 / 457,38 / 73 / 113 / 115 / 117 / 118 / 127 / 152 / 154 / 160 / 163 / 164 / 168 / 169 / 172 / 184 / 192 / 198 / 220 / 243 / 260 / 266 / 270 / 295 / 31 7 / 333 / 392 / 423,38 / 73 / 113 / 115 / 118 / 127 / 152 / 154 / 160 / 162 / 164 / 168 / 172 / 184 / 192 / 198 / 220 / 243 / 260 / 266 / 270 / 295 / 317 / 333 / 42 3,38 / 73 / 113 / 115 / 118 / 127 / 152 / 154 / 160 / 163 / 164 / 168 / 169 / 172 / 184 / 192 / 198 / 220 / 243 / 260 / 266 / 270 / 295 / 317 / 33 3 / 423,38 / 73 / 113 / 115 / 118 / 127 / 152 / 154 / 160 / 164 / 168 / 172 / 184 / 189 / 192 / 198 / 220 / 243 / 260 / 266 / 270 / 295 / 317 / 333 / 423,38 / 73 / 113 / 115 / 118 / 127 / 152 / 154 / 160 / 164 / 168 / 172 / 184 / 192 / 198 / 210 / 220 / 243 / 260 / 266 / 270 / 295 / 317 / 333 / 42 3, 38 / 73 / 113 / 115 / 118 / 127 / 152 / 154 / 160 / 164 / 168 / 172 / 184 / 192 / 198 / 220 / 243 / 260 / 266 / 270 / 295 / 317 / 333 / 423, 38 / 73 / 113 / 118 / 127 / 152 / 154 / 160 / 164 / 168 / 172 / 184 / 192 / 198 / 220 / 243 / 260 / 266 / 270 / 295 / 317 / 333 / 423, 38 / 73 / 113 / 118 / 127 / 152 / 154 / 160 / 164 / 168 / 172 / 184 / 192 / 198 / 220 / 243 / 266 / 270 / 317 / 333 / 423, 38 / 73 / 113 / 118 / 127 / 152 / 154 / 160 / 164 / 168 / 172 / 184 / 192 / 198 / 220 / 266 / 270 / 317 / 333 / 423, and38 / 113 / 127 / 152 / 154 / 160 / 164 / 168 / 172 / 198 / 220 / 290 / 317 / 423 and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 972.

130. The method of any one of Claims 75-80, wherein the reference sequence is SEQ ID NO. 5704 and the at least one substitution or substitution set comprises substitutions at amino acid positions selected from 48, 116, 118, 120, 130, 137, 153, 157, 163, 175, 181 , 187, 192, 213, 246, 263, 273, 296, 318, 334, 341 , 394, 412, 424, 454, and 458, and / or any combinations thereof, wherein the amino acid positions are numbered with reference to SEQ ID NO: 5704.131 . The method of any of Claims 75-130, wherein the engineered transposase comprises a polypeptide sequence selected from the even-numbered sequences selected from SEQ ID NOs: 2-2368, 2388-5694, 5702, and 5706-5756.

132. The method of any one of Claims 75-80 and 131 , wherein the engineered transposase comprises a polypeptide sequence of SEQ ID NO: 2396.

133. The method of any one of Claims 75-80 and 131 , wherein the engineered transposase comprises a polypeptide sequence of SEQ ID NO: 4642.

134. The method of any one of Claims 75-80 and 131 , wherein the engineered transposase comprises a polypeptide sequence of SEQ ID NO: 4872.

135. The method of any one of Claims 75-80 and 131 , wherein the engineered transposase comprises a polypeptide sequence of SEQ ID NO: 4954.

136. The method of any one of Claims 75-80 and 131 , wherein the engineered transposase comprises a polypeptide sequence of SEQ ID NO: 5186.

137. The method of any one of Claims 75-80 and 131 , wherein the engineered transposase comprises a polypeptide sequence of SEQ ID NO: 5354.

138. The method of any one of Claims 75-80 and 131 , wherein the engineered transposase comprises a polypeptide sequence of SEQ ID NO: 5500.

139. The method of any one of Claims 75-80 and 131 , wherein the engineered transposase comprises a polypeptide sequence of SEQ ID NO: 5520.

140. The method of any one of Claims 75-80 and 131 , wherein the engineered transposase comprises a polypeptide sequence of SEQ ID NO: 5702.141 . The method of any one of Claims 75-80 and 131 , wherein the engineered transposase comprises a polypeptide sequence of SEQ ID NO: 5738.

142. The method of any one of Claims 75-141 , wherein the engineered transposase is purified.

143. The method of any one of Claims 76-141 , wherein fragmenting the target nucleic acid comprises tagmentation or random sheering and adapter ligation.

144. The method of any one of Claims 76-141 , wherein fragmenting the target nucleic acid comprises tagmentation.

145. The method of Claim 75, wherein each altered nucleic acid of the mixture of altered nucleic acids comprises an identifiable sequence tag (1ST).

146. The method of any one of Claims 75-79, wherein each nucleic acid fragment of the subset of nucleic acid fragments comprises an identifiable sequence tag (1ST).

147. The method of Claim 155 or 156, wherein the 1ST is between 6 and 30 nucleotides in length.

148. The method of Claim 75, wherein each altered nucleic acid of the mixture of altered nucleic acids is between 30 and 30000 nucleotides in length.

149. The method of any one of Claims 76-79, wherein each nucleic acid fragment of the plurality of nucleic acid fragments is between 30 and 30000 nucleotides in length.

150. The method of any one of Claims 75-134, wherein the transposition event occurs in reaction conditions additionally comprising a terminal deoxynucleotide transferase, dNTP, and / or buffer components suitable for the addition of deoxynucleotides to the 3’ terminus of each of the plurality of nucleic acid fragments.151 . The method of any one of Claims 75-144, wherein the transposition event occurs in reaction conditions additionally comprising a DNA ligase and / or buffer components suitable for a ligation reaction.

152. The method of Claim 75, wherein each altered nucleic acid of the mixture of altered nucleic acids further comprises a sample tag and / or a unique molecular identifier (UMI).

153. The method of any one of Claims 76-79, wherein each nucleic acid fragment of the subset of nucleic acid fragments further comprises a sample tag and / or a unique molecular identifier (UMI).

154. The method of any one of Claims 77-144, wherein amplification of the subset of nucleic acid fragments is performed through polymerase chain reaction (PCR), multiple displacement amplification (MDA), ligase chain reaction (LCR), loop mediated isothermal amplification (LAMP), rolling circle amplification (RCA), or strand displacement amplification (SDA).

155. The method of any one of Claims 78-144, wherein the sequencing comprises nextgeneration sequencing (NGS).

156. The method of any one of Claims 78-144, wherein sequencing comprises sequencing by synthesis, sequencing by ligation, or nanopore sequencing.

157. The method of Claim 156, wherein the sequencing by synthesis comprises Illumina™ dye sequencing, single-molecule real-time (SMRT™) sequencing, or pyrosequencing.

158. The method of Claim 156, wherein the sequencing by ligation comprises polony-based sequencing or SOLiD™ sequencing.

159. The method of any one of Claims 75-158, wherein the target nucleic acid comprises genomic DNA or cDNAs from a single cell.

160. The method of any one of Claims 75-158, wherein the target nucleic acid comprises nucleic acids from a plurality of haplotypes.161 . The method of any one of Claims 75-158, wherein the target nucleic acid is crosslinked via histones or chromatin from single or multiple cells.

162. The method of any one of Claims 75-158, wherein the target nucleic acid has been condensed or optionally treated with one or more condensing agents.

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