Optimized Protein Linkers and Methods of Use

Abstract

The present invention relates to peptide linkers and fusion proteins comprising linkers designed to optimize the activity of the proteins contained therein, as well as methods for using the same. The present invention further relates to newly designed Cas12a-based cytosine base editors.

Description

[0001] Statement Regarding Electronic Submission of Sequence Listing

[0002] Instead of a paper copy, a sequence listing in ASCII text format was provided, which was submitted in accordance with 37 C.F.R.§1.821, named 1499-6WO_ST25.txt, sized 727,526 bytes, generated on July 17, 2020, and submitted via EFS-Web. This sequence listing is hereby incorporated by reference herein in that its disclosure is mentioned in the specification.

[0003] Priority Claim

[0004] This application claims the benefit of U.S. Provisional Application No. 62 / 876,275, filed on July 19, 2019 (the entire content of which is incorporated by reference herein), under 35 U.S.C.§119(e). Field of the Invention

[0005] The present invention relates to peptide linkers and fusion proteins comprising linkers designed to optimize the activity of the proteins contained therein, and methods for using the same. The present invention further relates to newly designed Cas12a-based cytosine base editors. Background of the Invention

[0007] In the past six years, CRISPR-based gene editing tools (especially those based on Cas9) have become increasingly popular. While early tools relied on the ability of Cas9 to generate blunt-ended double-strand breaks in DNA and double-strand break repair mechanisms such as homologous recombination and non-homologous end joining, more recent methods have been developed that primarily use modified versions of the nuclease as targeting tools for other covalently linked effector proteins. Notably, the first Cas9-based base editor was developed by linking Cas9 to a deaminase domain (see, e.g., Gaudelli et al., Nature 551:464-471 (2017)). The initial cytosine base editor was created by linking the rat APOBEC1 domain (apolipoprotein B mRNA editing enzyme), which deaminates cytosine to uracil in both RNA and DNA, to the N-terminus of Cas9 using a linker based on the previously published unstructured XTEN protein (Komor et al., Nature 533(7603):420-424 (2016)). A uracil-DNA glycosylase inhibitor (UGI) domain was linked to the C-terminus of Cas9 to reduce base excision repair activity. Later versions of the Cas9 cytosine base editor (CBE) doubled the length of both linkers by adding flexible glycine and serine residues and added additional UGI domains. The most recent versions of this base editor have been optimized for use in human cells through codon optimization and improved nuclear localization signals as well as ancestral reconstruction of the deaminase domain.

[0008] Cas12a (also known as Cpf1) is a more recently discovered CRISPR endonuclease that has also increasingly been used as a genome editing tool. Cas12a differs from Cas9 in several respects, including, for example, its size, its nuclease activity, the structure of the guide RNA, the orientation by which the nuclease binds its guide RNA, and the protospacer adjacent motif (PAM) that is recognized. While some variations of Cas12a-based cytosine base editors have been tested, they have lower activity compared to Cas9-based versions. Thus, to overcome the drawbacks in the art, new adenosine base editing tools using Cas12a are needed. SUMMARY OF THE INVENTION

[0010] Existing CRISPR-based cytosine base editors are severely Cas9-based, and the published versions of Cas12a-based cytosine base editors are relatively ineffective compared to Cas9-based versions. Part of this deficiency may be attributed to the different architecture and binding orientation of Cas12a compared to Cas9. Cas9-based cytosine base editors rely on simple GS linkers or previously designed unstructured sequences, and their length and composition have been designed for the optimal arrangement of the deaminase and UGI domains relative to the DNA being edited. In addition, base editors derived from Cas12a have not achieved an activity level suitable for commercial applications. The present inventors have designed novel linker sequences and optimized domain architecture for Cas12a-based cytosine base editors, which can now allow targeting of new sites and / or expanding the repertoire of site-specific base editing tools, and / or which may be suitable for commercial use. Methods of modifying nucleic acids by using the fusion proteins and / or polynucleotides encoding them of the present invention are also provided. These editors can be used for prokaryotic and / or eukaryotic applications, including editing the genomes of commercially relevant crops.

[0011] One aspect of the invention provides a polypeptide comprising any one of the amino acid sequences of SEQ ID NOs: 1-24 (L1-L24).

[0012] A second aspect of the invention provides a polypeptide comprising a Cas12a domain and any one of the amino acid sequences of SEQ ID NOs: 1-24.

[0013] A third aspect of the invention provides a fusion protein comprising a Cas12a domain, a polypeptide of interest, and any one of the amino acid sequences of SEQ ID NOs: 1-24.

[0014] The fourth aspect provides a Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-associated (Cas) (CRISPR-Cas) system, which comprises: (a) a fusion protein comprising a Cas12a domain, a linker comprising an amino acid sequence of any one of SEQ ID NOs: 1-24, and a polypeptide of interest, wherein the Cas12a domain is linked to the polypeptide of interest via any one of the amino acid sequences of SEQ ID NOs: 1-24; or a nucleic acid encoding the fusion protein; and (b) a guide nucleic acid (CRISPR RNA, CRISPR DNA, crRNA, crDNA, gRNA) comprising a spacer sequence and a repeat sequence, wherein the guide nucleic acid is capable of forming a complex with the Cas12a domain of the fusion protein, and the spacer sequence is capable of hybridizing with a target nucleic acid, thereby guiding the Cas12a domain and the polypeptide of interest to the target nucleic acid, whereby the system is capable of modifying or regulating the target nucleic acid.

[0015] The fifth aspect of the present invention provides a fusion protein, which comprises: (a) a Cas12a domain, wherein when associated with a bound guide nucleic acid (e.g., gRNA), the Cas12a domain specifically binds to a target nucleic acid sequence; (b) a cytidine deaminase domain, wherein when associated with the Cas12a domain and the gRNA, the cytidine deaminase domain deaminates cytosine bases in the single-stranded portion of the target nucleic acid sequence; and (c) a uracil glycosylase inhibitor (UGI) domain, wherein the UGI domain inhibits uracil-DNA glycosylase, wherein the Cas12a domain is linked to the cytidine deaminase domain or the UGI domain via any one of the amino acid sequences of SEQ ID NOs: 1-24.

[0016] The sixth aspect provides a fusion protein comprising: (a) a cytosine deaminase domain; (b) a Cas12a domain; and (c) a uracil-DNA glycosylase inhibitor (UGI) domain, wherein the C-terminus of the cytosine deaminase domain is linked to the N-terminus of the Cas12a domain via any one of the amino acid sequences of SEQ ID NOs: 1-5, and the C-terminus of the Cas12a domain is linked to the N-terminus of the UGI domain, or the C-terminus of the Cas12a domain is linked to the N-terminus of the UGI domain via any one of the amino acid sequences of SEQ ID NOs: 6-9, and the C-terminus of the cytosine deaminase domain is linked to the N-terminus of the Cas12a domain.

[0017] The seventh aspect provides a fusion protein comprising: (a) a Cas12a (Cpf1) domain; (b) a uracil-DNA glycosylase inhibitor (UGI) domain; and (c) a cytosine deaminase domain, wherein the C-terminus of the Cas12a domain is linked to the N-terminus of the UGI domain via any one of the amino acid sequences of SEQ ID NOs: 10-12, and the C-terminus of the UGI domain is linked to the N-terminus of the cytosine deaminase domain via any one of the amino acid sequences of SEQ ID NOs: 13-16, wherein the amino acid sequences of SEQ ID NOs: 10-12 and SEQ ID NOs: 13-16 are independently selected.

[0018] The eighth aspect provides a fusion protein comprising: (a) a uracil-DNA glycosylase inhibitor (UGI) domain; (b) a Cas12a (Cpf1) domain, wherein the Cas12a domain contains a mutation at the nuclease active site; and (c) a cytosine deaminase domain, wherein the C-terminus of the UGI domain is linked to the N-terminus of the Cas12a domain via any one of the amino acid sequences of SEQ ID NOs: 17-19, and the C-terminus of the Cas12a domain is linked to the N-terminus of the cytosine deaminase domain, or wherein the C-terminus of the UGI domain is linked to the N-terminus of the Cas12a domain, and the C-terminus of the Cas12a domain is linked to the N-terminus of the cytosine deaminase domain via any one of the amino acid sequences of SEQ ID NOs: 20-24.

[0019] The ninth aspect of the present invention provides a method for modifying a target nucleic acid, the method comprising contacting the target nucleic acid with the following items: (a)(i) the fusion protein of the present invention, and (a)(ii) a guide nucleic acid; (b) a complex comprising the fusion protein of the present invention and a guide nucleic acid; (c) a composition comprising the fusion protein of the present invention and a guide nucleic acid; and / or (d) the system of the present invention, thereby modifying the target nucleic acid.

[0020] The tenth aspect of the present invention provides a method for modifying a target nucleic acid, the method comprising contacting a cell or cell-free system comprising the target nucleic acid with the following items under certain conditions: (a)(i) a polynucleotide encoding the polypeptide or fusion protein of the present invention, or an expression cassette or vector comprising the same, and (a)(ii) a guide nucleic acid, or an expression cassette or vector comprising the same; and / or (b) a nucleic acid construct encoding a complex comprising the fusion protein of the present invention and a guide nucleic acid, or an expression cassette or vector comprising the same, thereby modifying the target nucleic acid, the conditions being such that when the fusion protein is expressed and forms a complex with the guide nucleic acid, the complex hybridizes to the target nucleic acid.

[0021] The eleventh aspect of the present invention provides a method for editing a target nucleic acid, the method comprising contacting the target nucleic acid with the following items: (a)(i) the fusion protein of the present invention, and (a)(ii) a guide nucleic acid; (b) a complex comprising the fusion protein of the present invention and a guide nucleic acid; (c) a composition comprising (i) the fusion protein of the present invention and (ii) a guide nucleic acid; and / or (d)(i) the system of the present invention, wherein the cytosine deaminase domain converts cytosine (C) in the target nucleic acid to thymine (T), thereby editing the target nucleic acid to produce a (point) mutation.

[0022] The twelfth aspect of the present invention provides a method for editing a target nucleic acid, the method comprising contacting a cell or cell-free system comprising the target nucleic acid with the following items under certain conditions: (a)(i) a polynucleotide encoding the fusion protein of the present invention, or an expression cassette or vector comprising the same, and (a)(ii) a guide nucleic acid, or an expression cassette or vector comprising the same; and / or (b) a nucleic acid construct encoding a complex comprising the fusion protein of the present invention and a guide nucleic acid, or an expression cassette or vector comprising the same, the conditions being such that when the fusion protein is expressed and forms a complex with the guide nucleic acid, the complex hybridizes to the target nucleic acid, wherein the cytosine deaminase domain converts cytosine (C) in the target nucleic acid to thymine (T), thereby editing the target nucleic acid to produce a (point) mutation.

[0023] The present invention further provides constructs, complexes, compositions, expression cassettes, vectors and cells, which comprise the polypeptides and / or fusion proteins of the present invention, and / or polynucleotides and nucleic acid constructs encoding the fusion proteins and complexes of the present invention.

[0024] These and other aspects of the present invention are set forth in more detail in the following description of the present invention.

[0025] Brief Description of Sequences

[0026] SEQ ID NO: 1-24 are the amino acid sequences of the present invention useful for linker polypeptides.

[0027] SEQ ID NO: 25-28 are the amino acid sequences of exemplary peptide linkers useful for linker polypeptides.

[0028] SEQ ID NO: 29-45 are exemplary Cas12a amino acid sequences that can be used in the present invention.

[0029] SEQ ID NO: 46-47 and SEQ ID NO: 76-82 are exemplary cytosine deaminase amino acid sequences that can be used in the present invention.

[0030] SEQ ID NO: 48 is an exemplary uracil-DNA glycosylase inhibitor (UGI).

[0031] SEQ ID NO: 49-72 and SEQ ID NO: 91-107 are exemplary fusion proteins.

[0032] SEQ ID NO: 83-88 are exemplary spacer sequences.

[0033] SEQ ID NO: 89 and SEQ ID NO: 90 are exemplary intron sequences of human and soybean, respectively. Brief Description of Drawings

[0035] Figure 1 A-C provide exemplary domain arrangements of the Cas12a-based cytosine base editors of the present invention selected for experimental screening in mammalian cells. For the constructs in Figure 1 A (ACU) and Figure 1 C (UCA), each linker with APOBEC1 or UGI was independently tested and paired with a control linker (8-residue GS linker, XTEN linker or GS-XTEN-GS linker). For the constructs in Figure 1 B (CUA), all combinations of linkers were tested.

[0036] Figure 2 Two Cas12a cytosine base editors are provided for use as a reference.

[0037] Figure 3 The results of C to T editing using the EMX1 spacer 1: TCATCTGTGCCCCTCCCTCCCTG (SEQ ID NO: 83) are shown. The Y-axis indicates the level of C->T editing observed for a given cytosine at a specific position within the spacer.

[0038] Figure 4 The results of C to T editing using the RUNX1 spacer 1: AGCCTCACCCCTCTAGCCCTACA (SEQ ID NO: 84) are shown. The Y-axis indicates the level of C->T editing observed for a given cytosine at a specific position within the spacer.

[0039] Figure 5 The results of C to T editing using the RUNX1 spacer 2: TTCTCCCCTCTGCTGGATACCTC (SEQ ID NO: 85) are shown. The Y-axis indicates the level of C->T editing observed for a given cytosine at a specific position within the spacer. No editing data is available for constructs UCA_L2_1, UCA_L2_1R, UCA_L2_4, CUA control, or Shanghai Tech control.

[0040] Figure 6 The results of C to T editing using the DNMT1 spacer 1: CCTCACTCCTGCTCGGTGAATTT (SEQ ID NO: 86) are shown. The Y-axis indicates the level of C->T editing observed for a given cytosine at a specific position within the spacer.

[0041] Figure 7 The results of C to T editing using the DNMT1 spacer 2: GCTCAGCAGGCACCTGCCTCAGC (SEQ ID NO: 87) are shown. The Y-axis indicates the level of C->T editing observed for a given cytosine at a specific position within the spacer. No editing data is available for construct ACU_L1_1.

[0042] Figure 8 The results of C to T editing using the EMX1 spacer 1: TCATCTGTGCCCCTCCCTCCCTG (SEQ ID NO: 83) are shown. The Y-axis indicates the level of C->T editing observed for a given cytosine at a specific position within the spacer.

[0043] Figure 9 Shows the results of C to T editing using RUNX1 spacer 1: AGCCTCACCCCTCTAGCCCTACA (SEQ ID NO: 84).

[0044] Figure 10 Shows the results of C to T editing using DNMT1 spacer 1: CCTCACTCCTGCTCGGTGAATTT (SEQ ID NO: 86). The Y-axis indicates the level of C->T editing observed for a given cytosine at a specific position within the spacer. No editing data is available for constructs ACU_L1_2, ACU_L2_2R.

[0045] Figure 11 Shows the results of C to T editing using DNMT1 spacer 2: GCTCAGCAGGCACCTGCCTCAGC (SEQ ID NO: 87). The Y-axis indicates the level of C->T editing observed for a given cytosine at a specific position within the spacer.

[0046] Figure 12 Shows the results of C to T editing using EMX1 spacer 1: TCATCTGTGCCCCTCCCTCCCTG (SEQ ID NO: 83). The Y-axis indicates the level of C->T editing observed for a given cytosine at a specific position within the spacer.

[0047] Figure 13 Shows the results of C to T editing using RUNX1 spacer 1: AGCCTCACCCCTCTAGCCCTACA (SEQ ID NO: 84). The Y-axis indicates the level of C->T editing observed for a given cytosine at a specific position within the spacer.

[0048] Figure 14 Shows the results of C to T editing using RUNX1 spacer 2: TTCTCCCCTCTGCTGGATACCTC (SEQ ID NO: 85). The Y-axis indicates the level of C->T editing observed for a given cytosine at a specific position within the spacer. No editing data is available for constructs ACU_L1_1, ACU_L1_2, ACU_L1_3, ACU_L1_3R, ACU_L1_5R, CUA_L1_3_L2_1, UCA_L1_1, UCA_L2_1, UCA_L2_1R, and UCA_L2_4.

[0049] Figure 15Shows the results of C-to-T editing using the AAVS1 spacer 1: TCTGTCCCCTCCACCCCACAGTG (SEQ ID NO: 88). The Y-axis indicates the level of C->T editing observed for a given cytosine at a specific position within the spacer.

[0050] Figure 16 Shows the results of C-to-T editing using the DNMT1 spacer 1: CCTCACTCCTGCTCGGTGAATTT (SEQ ID NO: 86). The Y-axis indicates the level of C->T editing observed for a given cytosine at a specific position within the spacer.

[0051] Figure 17 Shows the results of C-to-T editing using the EMX1 spacer 1: TCATCTGTGCCCCTCCCTCCCTG (SEQ ID NO: 83). The Y-axis indicates the level of C->T editing observed for a given cytosine at a specific position within the spacer.

[0052] Figure 18 Shows the results of C-to-T editing using the RUNX1 spacer 1: AGCCTCACCCCTCTAGCCCTACA (SEQ ID NO: 84). The Y-axis indicates the level of C->T editing observed for a given cytosine at a specific position within the spacer.

[0053] Figure 19 Shows the results of C-to-T editing using the AAVS1 spacer 1: TCTGTCCCCTCCACCCCACAGTG (SEQ ID NO: 88). The Y-axis indicates the level of C->T editing observed for a given cytosine at a specific position within the spacer.

[0054] Figure 20 Shows the results of C-to-T editing using the DNMT1 spacer 1: CCTCACTCCTGCTCGGTGAATTT (SEQ ID NO: 86). The Y-axis indicates the level of C->T editing observed for a given cytosine at a specific position within the spacer. For the construct ACU_L1_2 (A3A), no editing data is available.

[0055] Figure 21 Shows the results of C-to-T editing using the EMX1 spacer 1: TCATCTGTGCCCCTCCCTCCCTG (SEQ ID NO: 83). The Y-axis indicates the level of C->T editing observed for a given cytosine at a specific position within the spacer.

[0056] Figure 22 Shows the results of C to T editing using the AAVS1 spacer 1: TCTGTCCCCTCCACCCCACAGTG (SEQ ID NO: 88). The Y-axis indicates the level of C->T editing observed for a given cytosine at a specific position within the spacer.

[0057] Figure 23 Shows the results of C to T editing using the RUNX1 spacer 1: AGCCTCACCCCTCTAGCCCTACA (SEQ ID NO: 84). The Y-axis indicates the level of C->T editing observed for a given cytosine at a specific position within the spacer. No editing data (ND) was available for the constructs UC_L2_1 (A3A + intron) and the ACU control.

[0058] Figure 24 Shows the results of C to T editing using the RUNX1 spacer 2: TTCTCCCCTCTGCTGGATACCTC (SEQ ID NO: 85). The Y-axis indicates the level of C->T editing observed for a given cytosine at a specific position within the spacer. No editing data (ND) was available for the construct UC_L2_4.

[0059] Figure 25 Shows the results of C to T editing using the DNMT1 spacer 1: CCTCACTCCTGCTCGGTGAATTT (SEQ ID NO: 86). The Y-axis indicates the level of C->T editing observed for a given cytosine at a specific position within the spacer.

[0060] Figure 26 Shows the results of editing three different nucleic acid targets (locus 1, locus 2, locus 3) in soybean using the editor constructs of the present invention, as described in Example 4. DETAILED DESCRIPTION OF THE INVENTION

[0062] Now, the present invention will be described hereinafter with reference to the accompanying drawings and embodiments (in which embodiments of the present invention are shown). This description is not intended to be an exhaustive cataloging of all the different ways in which the present invention can be practiced or of all the features that can be added to the present invention. For example, features illustrated with respect to one embodiment can be incorporated into other embodiments, and features illustrated with respect to a particular embodiment can be deleted from that embodiment. Thus, the present invention contemplates that in some embodiments of the present invention, any feature or combination of features set forth herein can be excluded or omitted. Additionally, in light of the present disclosure, numerous variations and additions to the various embodiments herein will be apparent to those of ordinary skill in the art, which do not depart from the present invention. Accordingly, the following description is intended to illustrate some particular embodiments of the present invention, rather than to exhaustively detail all of its permutations, combinations, and variations.

[0063] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terms used in the description of the invention herein are for the purpose of describing particular embodiments only and are not intended to limit the present invention.

[0064] All publications, patent applications, patents, and other references cited herein are incorporated by reference in their entirety for the teachings relevant to the sentence and / or paragraph in which the reference is presented.

[0065] Unless the context dictates otherwise, it is specifically intended that the various features of the present invention described herein can be used in any combination. Additionally, the present invention also contemplates that in some embodiments of the present invention, any feature or combination of features set forth herein can be excluded or omitted. By way of illustration, if the specification states that a composition comprises components A, B, and C, then it is specifically intended that any one of A, B, or C, or any combination thereof, can be omitted and disclaimed, either singly or in any combination.

[0066] As used in the description of the present invention and the appended claims, the singular forms "a", "an", and "the" are intended to include the plural forms as well, unless the context clearly dictates otherwise.

[0067] Furthermore, as used herein, "and / or" refers to and encompasses any and all possible combinations of one or more of the associated listed items, as well as the lack of combinations when interpreted in terms of alternatives ("or").

[0068] As used herein, when referring to measurable values such as amounts or concentrations, etc., the term "about" means encompassing variations of ±10%, ±5%, ±1%, ±0.5%, or even ±0.1% of the specified value, as well as the specified value. For example, "about X" (where X is a measurable value) means including X, as well as variations of ±10%, ±5%, ±1%, ±0.5%, or even ±0.1% of X. The ranges provided herein for measurable values can include any other ranges and / or individual values therein.

[0069] As used herein, phrases such as "between X and Y" and "between about X and Y" shall be construed to include X and Y. As used herein, phrases such as "between about X and Y" mean "between about X and about Y", and phrases such as "from about X to Y" mean "from about X to about Y".

[0070] As used herein, the terms "comprising", "containing", and "including" expressly state the presence of the recited features, integers, steps, operations, elements, and / or components, but do not preclude the presence or addition of one or more other features, integers, steps, operations, elements, components, and / or groups thereof.

[0071] As used herein, the transitional phrase "consisting essentially of" means that the scope of the claim will be construed to cover the specified materials or steps recited in the claim, as well as those that do not materially affect the basic and novel features of the claimed invention. Thus, when used in the claims of the present invention, the term "consisting essentially of" is not intended to be construed as equivalent to "comprising".

[0072] As used herein, the terms "increase" and "enhance" (and their grammatical variations) describe an improvement of at least about 25%, 50%, 75%, 100%, 150%, 200%, 300%, 400%, 500%, or more compared to a control.

[0073] As used herein, the terms "reduce", "decrease", and "lower" (and their grammatical variations) describe, for example, a reduction of at least about 5%, 10%, 15%, 20%, 25%, 35%, 50%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100% compared to a control. In particular embodiments, the reduction may result in no or substantially no (i.e., a non-significant amount, such as less than about 10% or even 5%) detectable activity or amount.

[0074] A "heterologous" or "recombinant" nucleotide sequence is a nucleotide sequence that is not naturally associated with the host cell into which it is introduced, including non-naturally occurring multiple copies of a naturally occurring nucleotide sequence.

[0075] A "native" or "wild-type" nucleic acid, nucleotide sequence, polypeptide, or amino acid sequence refers to a nucleic acid, nucleotide sequence, polypeptide, or amino acid sequence that occurs naturally or is endogenous. Thus, for example, "wild-type mRNA" is mRNA that occurs naturally or is endogenous to an organism. A "homologous" nucleic acid sequence is a nucleotide sequence that is naturally associated with the host cell into which it is introduced.

[0076] As used herein, the terms "nucleic acid", "nucleic acid molecule", "nucleotide sequence", and "polynucleotide" refer to linear or branched, single-stranded or double-stranded RNA or DNA, or a hybrid thereof. The term also encompasses RNA / DNA hybrids. When producing dsRNA synthetically, less common bases (e.g., inosine, 5-methylcytosine, 6-methyladenine, hypoxanthine, and others) can also be used for antisense dsRNA and ribozyme pairing. For example, polynucleotides containing C-5 propyne analogs of uridine and cytidine have been shown to bind RNA with high affinity and are potent antisense inhibitors of gene expression. Other modifications can also be made, such as modifications to the phosphodiester backbone of RNA or to the 2'-hydroxyl group in the ribose moiety.

[0077] As used herein, the term "nucleotide sequence" refers to a heteropolymer of nucleotides, or the order of these nucleotides from the 5' to the 3' end of a nucleic acid molecule, and includes DNA or RNA molecules, including cDNA, DNA fragments or portions, genomic DNA, synthetic (e.g., chemically synthesized) DNA, plasmid DNA, mRNA, and antisense RNA, any of which may be single-stranded or double-stranded. The terms "nucleotide sequence", "nucleic acid", "nucleic acid molecule", "oligonucleotide", and "polynucleotide" are also used interchangeably herein to refer to a heteropolymer of nucleotides. The nucleic acid molecules and / or nucleotide sequences provided herein are presented herein from left to right in the 5' to 3' direction and are represented using the standard codes for representing nucleotide characters as set forth in U.S. Sequence Rules 37 CFR §§ 1.821-1.825 and World Intellectual Property Organization (WIPO) Standard ST.25. As used herein, the "5' region" may mean the region of a polynucleotide that is closest to the 5' end of the polynucleotide. Thus, for example, an element in the 5' region of a polynucleotide may be located anywhere from the first nucleotide at the 5' end of the polynucleotide to a nucleotide midway through the entire polynucleotide. As used herein, the "3' region" may mean the region of a polynucleotide that is closest to the 3' end of the polynucleotide. Thus, for example, an element in the 3' region of a polynucleotide may be located anywhere from the first nucleotide at the 3' end of the polynucleotide to a nucleotide midway through the entire polynucleotide.

[0078] As used herein, the term "gene" refers to a nucleic acid molecule capable of being used to produce mRNA, antisense RNA, miRNA, anti - microRNA antisense oligodeoxynucleotide (AMO), etc. A gene may or may not be capable of being used to produce a functional protein or gene product. A gene may include both coding and non - coding regions (e.g., introns, regulatory elements, promoters, enhancers, termination sequences, and / or 5' and 3' untranslated regions). A gene may be "isolated", meaning such a nucleic acid that is substantially or essentially free of components normally found associated with the nucleic acid in its native state. Such components include other cellular materials, media from recombinant production, and / or various chemicals used in chemically synthesizing the nucleic acid.

[0079] By way of example, introns useful in the constructs of the present invention include, but are not limited to, SEQ ID NO:89 or SEQ ID NO:90.

[0080] The term "mutation" refers to point mutations (e.g., missense, nonsense, or insertion or deletion of a single base pair that results in a frameshift), insertions, deletions, and / or truncations. When a mutation is a substitution of one residue for another within an amino acid sequence or a deletion or insertion of one or more residues within the sequence, the mutation is typically described by identifying the original residue, followed by the position of that residue within the sequence and the identity of the newly substituted residue.

[0081] As used herein, the term "complementary" or "complementarity" refers to the natural binding of polynucleotides through base pairing under permissive salt and temperature conditions. For example, the sequence "A-G-T" (5' to 3') binds to the complementary sequence "T-C-A" (3' to 5'). Complementarity between two single-stranded molecules can be "partial", where only some of the nucleotides bind, or it can be complete, when there is full complementarity between the single-stranded molecules. The degree of complementarity between nucleic acid strands has a significant effect on the efficiency and strength of hybridization between the nucleic acid strands.

[0082] As used herein, "complement" can mean 100% complementarity to a comparator nucleotide sequence, or it can mean less than 100% complementarity (e.g., complementarity of about 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, etc.).

[0083] A "portion" or "fragment" of a nucleotide sequence of the present invention will be understood to mean a nucleotide sequence that has a reduced length (e.g., reduced by 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more nucleotides) relative to a reference nucleic acid or nucleotide sequence, and that comprises, consists essentially of, and / or consists of a nucleotide sequence of contiguous nucleotides that is identical or nearly identical (e.g., 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identical) to the reference nucleic acid or nucleotide sequence. When appropriate, such nucleic acid fragments or portions according to the present invention may be included in a larger polynucleotide of which they are a component. As an example, the repeat sequences of the guide nucleic acids of the present invention may comprise a portion of a wild-type Cas12a repeat sequence.

[0084] As used herein with respect to polypeptides, the terms "fragment" or "portion" can refer to a polypeptide that has a reduced length relative to a reference polypeptide, and that comprises, consists essentially of, and / or consists of an amino acid sequence of contiguous amino acids that is identical or nearly identical (e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identical) to the corresponding portion of the reference polypeptide. When appropriate, such polypeptide fragments may be included in a larger polypeptide of which they are a component. In some embodiments, the polypeptide fragment comprises, consists essentially of, or consists of at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 125, 150, 175, 200, 225, 250, 260, 270, 280, 290 or more contiguous amino acid residues of the reference polypeptide.

[0085] Different nucleic acids or proteins having homology are referred to herein as "homologs". The term "homolog" includes homologous sequences from the same and other species, and orthologous sequences from the same and other species. "Homology" refers to the level of similarity (e.g., sequence similarity or identity) between two or more nucleic acid and / or amino acid sequences in terms of the percentage of positional identity. Homology also refers to the concept of similar functional properties among different nucleic acids or proteins. Thus, the compositions and methods of the present invention further comprise homologs of the nucleotide sequences and polypeptide sequences of the present invention. As used herein, "orthologous(ous)" refers to homologous nucleotide sequences and / or amino acid sequences in different species that have arisen from a common ancestral gene during speciation. Homologs of the nucleotide sequences of the present invention have substantial sequence identity (e.g., at least about 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or 100%) with the nucleotide sequences of the present invention.

[0086] As used herein, "sequence identity" refers to the extent to which two optimally aligned polynucleotide or polypeptide sequences are invariant throughout the aligned window of components (e.g., nucleotides or amino acids). "Identity" can be readily calculated by known methods, including but not limited to those described in the following: Computational Molecular Biology (Lesk, A.M., ed.) Oxford University Press, New York (1988); Biocomputing: Informatics and Genome Projects (Smith, D.W., ed.) Academic Press, New York (1993); Computer Analysis of Sequence Data, Part I (Griffin, A.M. and Griffin, H.G., eds.) Humana Press, New Jersey (1994); Sequence Analysis in Molecular Biology (von Heinje, G., ed.) Academic Press (1987); and Sequence Analysis Primer (Gribskov, M. and Devereux, J., eds.) Stockton Press, New York (1991).

[0087] As used herein, the term "percent sequence identity" or "identity percent" refers to the percentage of nucleotides that are identical in the linear polynucleotide sequence of a reference ("query") polynucleotide molecule (or its complementary strand) compared to a test ("subject") polynucleotide molecule (or its complementary strand) when the two sequences are aligned in an optimal manner. In some embodiments, "identity percent" may refer to the percentage of amino acids that are identical in an amino acid sequence compared to a reference polypeptide.

[0088] As used herein, in the context of two nucleic acid molecules, nucleotide sequences or protein sequences, the phrase "substantially identical" or "substantial identity" refers to two or more sequences or subsequences having at least about 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or 100% nucleotide or amino acid residue identity when compared and aligned for maximum correspondence, as measured by using one of the following sequence comparison algorithms or by visual inspection. In some embodiments of the invention, in a region of contiguous nucleotides of a nucleotide sequence of the invention that is about 10 nucleotides to about 30 nucleotides, about 15 nucleotides to about 25 nucleotides, about 30 nucleotides to about 40 nucleotides, about 50 nucleotides to about 60 nucleotides, about 70 nucleotides to about 80 nucleotides, about 90 nucleotides to about 100 nucleotides or more nucleotides in length, and any range therein up to the full length of the sequence, there is substantial identity. In some embodiments, the nucleotide sequence may be substantially identical over at least about 20 nucleotides (e.g., about 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40 nucleotides). In some embodiments, a substantially identical nucleotide or protein sequence performs substantially the same function as the nucleotide sequence (or the encoded protein sequence) to which it is substantially identical.

[0089] For sequence comparison, one sequence typically serves as a reference sequence, to which the test sequence is compared. When using a sequence comparison algorithm, the test and reference sequences are input into a computer, subsequence coordinates are designated (if necessary), and sequence algorithm program parameters are designated. The sequence comparison algorithm then calculates the percent sequence identity of the test sequence relative to the reference sequence based on the designated program parameters.

[0090] Optimal sequence alignments for comparing comparison windows are well known to those skilled in the art and can be performed using tools such as the local homology algorithms of Smith and Waterman, the homology alignment algorithms of Needleman and Wunsch, the similarity search methods of Pearson and Lipman, and optionally by computerized implementations of these algorithms such as GAP, BESTFIT, FASTA, and TFASTA (which are available as part of Wisconsin (Accelrys Inc., San Diego, CA)). The "identity score" for an aligned segment of a test sequence and a reference sequence is the number of identical components shared by the two aligned sequences divided by the total number of components in the reference sequence segment (e.g., the entire reference sequence or a smaller defined portion of the reference sequence). The percent sequence identity is expressed as the identity score multiplied by 100. The comparison of one or more polynucleotide sequences can be with a full-length polynucleotide sequence or a portion thereof, or with a longer polynucleotide sequence. For the purposes of the present invention, the "percent identity" can also be determined by using BLASTX version 2.0 (for translated nucleotide sequences) and BLASTN version 2.0 (for polynucleotide sequences).

[0091] Two nucleotide sequences are also considered to be substantially complementary when they hybridize to each other under stringent conditions. In some representative embodiments, two nucleotide sequences that are considered to be substantially complementary hybridize to each other under highly stringent conditions.

[0092] "Stringent hybridization conditions" and "stringent hybridization wash conditions" in the context of nucleic acid hybridization experiments such as Southern and Northern hybridizations are sequence-dependent and vary under different environmental parameters. Exhaustive guidelines for nucleic acid hybridization can be found in Tijssen Laboratory Techniques in Biochemistry and Molecular Biology - Hybridization with Nucleic Acid Probes, Chapter 2, Part I, "Overview of principles of hybridization and the strategy of nucleic acid probe assays", Elsevier, New York (1993). Generally, highly stringent hybridization and wash conditions are selected to be about 5 °C below the thermal melting point (T m ) for a particular sequence at a specified ionic strength and pH.

[0093] T m is the temperature at which 50% of the target sequence hybridizes to a perfectly matched probe (at a specified ionic strength and pH). Very stringent conditions are selected to be equal to T for a particular probe m . Examples of stringent hybridization conditions for hybridization of complementary nucleotide sequences having more than 100 complementary residues on a filter in Southern or Northern blots are 50% formamide with 1 mg of heparin at 42°C, where hybridization is carried out overnight. An example of highly stringent wash conditions is 0.15 M NaCl at 72°C for approximately 15 minutes. An example of stringent wash conditions is washing in 0.2x SSC at 65°C for 15 minutes (see Sambrook (below), see description of SSC buffer). Frequently, low stringency washes are carried out before high stringency washes to remove background probe signal. An example of moderately stringent wash conditions for a duplex of, for example, more than 100 nucleotides is 1x SSC at 45°C for 15 minutes. An example of low stringency wash conditions for a duplex of, for example, more than 100 nucleotides is 4 - 6x SSC at 40°C for 15 minutes. For short probes (e.g., approximately 10 to 50 nucleotides), stringent conditions typically involve a salt concentration of less than approximately 1.0 M Na ion at pH 7.0 to 8.3, typically a Na ion (or other salt) concentration of about 0.01 to 1.0 M, and the temperature is typically at least about 30°C. Stringent conditions can also be achieved by adding destabilizing agents such as formamide. Usually, a signal-to-noise ratio of 2x (or higher) compared to that observed for an unrelated probe in a particular hybridization assay indicates specific hybridization has been detected. Nucleotide sequences that do not hybridize to each other under stringent conditions are still substantially identical if the proteins they encode are substantially identical. This can occur, for example, when using the maximum codon degeneracy allowed by the genetic code to generate copies of a nucleotide sequence.

[0094] Any nucleotide sequence, polynucleotide, and / or recombinant nucleic acid construct of the present invention can be codon-optimized for expression in any target organism. Codon optimization is well-known in the art and involves modifying a nucleotide sequence for codon usage bias by using a species-specific codon usage table. The codon usage table is generated based on sequence analysis of the most highly expressed genes of the target organism / species. When the nucleotide sequence is to be expressed in the nucleus, the codon usage table is generated based on sequence analysis of the highly expressed nuclear genes of the target species. Modification of the nucleotide sequence is determined by comparing the species-specific codon usage table with the codons present in the native polynucleotide sequence. As understood in the art, codon optimization of a nucleotide sequence results in a nucleotide sequence that has less than 100% identity (e.g., 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 99.9% and any range or value therein) with the native nucleotide sequence but still encodes a polypeptide having the same function as the polypeptide encoded by the original native nucleotide sequence. Thus, in some embodiments of the present invention, the polynucleotides, nucleic acid constructs, expression cassettes, and / or vectors of the present invention (which contain / encode the polypeptides, fusion proteins, complexes of the present invention, such as Cas12a, target polypeptides, cytosine deaminases, linkers) can be codon-optimized for expression in a specific target species, such as a specific plant species, a specific bacterial species, a specific animal species, etc. In some embodiments, the codon-optimized polynucleotides, nucleic acid constructs, expression cassettes, and / or vectors of the present invention have about 70% to about 99.9% (e.g., 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%, or 100%) identity or more with the non-codon-optimized polynucleotides, nucleic acid constructs, expression cassettes, and / or vectors of the present invention.

[0095] In any of the embodiments described herein, the polynucleotides or nucleic acid constructs of the invention can be operably associated with a variety of promoters or other regulatory elements for expression in a target organism and / or in cells of a target organism. Thus, in some embodiments, an expression cassette or vector comprising a polynucleotide or nucleic acid construct of the invention can further comprise one or more promoters, enhancers, and / or terminators operably linked to one or more polynucleotides or nucleic acid constructs.

[0096] As used herein, "operably linked" or "operably associated" means that the indicated elements are functionally related and are generally also physically associated. Thus, as used herein, the terms "operably linked" or "operably associated" refer to nucleotide sequences on a single nucleic acid molecule that are functionally related. Thus, a first nucleotide sequence operably linked to a second nucleotide sequence means the case when the first nucleotide sequence is placed in a manner that has a functional relationship with the second nucleotide sequence. For example, a promoter is operably associated with a nucleotide sequence if the promoter affects the transcription or expression of the nucleotide sequence. One of ordinary skill in the art will appreciate that a control sequence (e.g., a promoter) need not be contiguous with the nucleotide sequence with which it is operably associated, so long as the control sequence functions to direct its expression. Thus, for example, intervening non-translated (yet transcribed) sequences can be present between a promoter and a nucleotide sequence, and the promoter can still be considered to be "operably linked" to the nucleotide sequence.

[0097] As used herein, with respect to polypeptides, the term "linked" means that one polypeptide is attached to another polypeptide. Polypeptides can be linked to another polypeptide (at the N-terminus or C-terminus) directly (e.g., via a peptide bond) or through a linker.

[0098] The term "linker" is well - recognized in the art and refers to a bond, chemical group, or molecule that connects two molecules or moieties (e.g., two domains of a fusion protein, such as a Cas12a domain and a nucleic acid editing domain (e.g., a cytosine deaminase)). The linker can be composed of a single linking molecule (e.g., an amino acid) or can contain more than one linking molecule. In some embodiments, the linker can be an organic molecule, group, polymer, or chemical moiety. In some embodiments, the linker can be an amino acid or peptide linker. In some embodiments, the length of the peptide linker can be about 4, 5 to 100 or more amino acids, such as 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100 or more amino acids. In some embodiments, the peptide linker can be a GS linker. In some embodiments, the linker can contain the amino acid sequence SGGS (SEQ ID NO:25), (GGS)n, or S(GGS)n (one or more repeats of SEQ ID NO:25), where n is 1 - 20 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 and any range or value therein). In some embodiments, the linker can contain the amino acid sequence SGGSGGSGGS (SEQ ID NO:26). In some embodiments, the linker can contain the amino acid sequence SGSETPGTSESATPES (SEQ ID NO:27), also known as the XTEN linker. In some embodiments, the linker can contain the amino acid sequence SGGSSGGSSGSETPGTSESATPESSGGSSGGS (SEQ ID NO:28), also known as the GS - XTEN - GS linker. In some embodiments, the linker comprises, consists essentially of, or consists of any one of the amino acid sequences of SEQ ID NO:1 - 24.

[0099] "Promoter" means a nucleotide sequence that controls or regulates the transcription of a nucleotide sequence (e.g., a coding sequence) operably associated with the promoter. The coding sequence controlled or regulated by the promoter can encode a polypeptide and / or functional RNA. Typically, a "promoter" refers to a nucleotide sequence that contains a binding site for RNA polymerase II and directs the initiation of transcription. Generally, a promoter is found 5' or upstream relative to the start point of the coding region of the corresponding coding sequence. The promoter region can contain other elements that function as regulators of gene expression. These include the TATA box consensus sequence, and often, the CAAT box consensus sequence (Breathnach and Chambon, (1981) Annu. Rev. Biochem. 50:349). In plants, the CAAT box can be replaced by the AGGA box (Messing et al., (1983) Genetic Engineering of Plants, T. Kosuge, C. Meredith and A. Hollaender (eds.), Plenum Press, pp. 211-227).

[0100] Promoters can include, for example, constitutive, inducible, temporally regulated, developmentally regulated, chemically regulated, tissue-preferred, and / or tissue-specific promoters for use in the preparation of recombinant nucleic acid molecules such as "synthetic nucleic acid constructs" or "protein-RNA complexes". These various types of promoters are known in the art.

[0101] The choice of promoter can vary depending on the temporal and spatial requirements for expression and can also vary based on the host cell to be transformed. Promoters for many different organisms are well known in the art. Based on the extensive knowledge available in the art, a suitable promoter can be selected for a particular host organism of interest. Thus, for example, much is known about the promoters upstream of genes that are highly constitutively expressed in model organisms, and such knowledge can be readily accessed and implemented (where appropriate) in other systems.

[0102] In some embodiments, the polynucleotides and / or nucleic acid constructs of the invention can be "expression cassettes" or can be contained within an expression cassette. As used herein, "expression cassette" means a recombinant nucleic acid molecule that contains, for example, a nucleic acid construct of the invention (e.g., that encodes a complex of the invention (e.g., a fusion protein and a guide nucleic acid of the invention)), wherein the nucleic acid construct is operably associated with at least one control sequence (e.g., a promoter). Thus, some embodiments of the invention provide expression cassettes that are designed for the expression of, for example, a nucleic acid construct of the invention.

[0103] An expression cassette comprising a nucleotide sequence of interest may be chimeric, meaning that at least one of its components is heterologous relative to at least one of its other components (e.g., a promoter from a host organism operably linked to a polynucleotide of interest to be expressed in the host organism, where the polynucleotide of interest is from a different organism than the host or is not normally found associated with that promoter). The expression cassette may also be an expression cassette that is naturally occurring but has been obtained in a recombinant form useful for heterologous expression.

[0104] The expression cassette may optionally include a transcriptional and / or translational termination region (i.e., a termination region) and / or an enhancer region that is functional in the selected host cell. A variety of transcriptional terminators and / or enhancers are available for use in the expression cassette and are responsible for termination of transcription and proper mRNA polyadenylation. The termination region and / or enhancer region may be native to the transcriptional initiation region, may be native to the nucleotide sequence of interest operably linked, may be native to the host cell, or may be from another source (e.g., foreign or heterologous to the promoter, nucleotide sequence of interest, host, or any combination thereof).

[0105] The expression cassette of the present invention may also include a nucleotide sequence encoding a selectable marker (which may be used to select transformed host cells). As used herein, "selectable marker" means a nucleotide sequence that, when expressed, confers a different phenotype on the host cell expressing the marker and thus allows such transformed cells to be distinguished from those that do not have the marker. Such nucleotide sequences may encode selectable or screenable markers, depending on whether the marker confers a trait that can be selected by chemical means, e.g., by using a selection reagent (e.g., an antibiotic, etc.), or depending on whether the marker is a trait that can be identified only by observation or testing, e.g., by screening (e.g., fluorescence). Many examples of suitable selectable markers are known in the art and may be used in the expression cassettes described herein.

[0106] In addition to expression cassettes, the nucleic acid molecules / constructs and polynucleotide sequences described herein can also be used in connection with vectors. The term "vector" refers to a composition for transferring, delivering, or introducing nucleic acids into cells. A vector contains a nucleic acid molecule having a nucleotide sequence to be transferred, delivered, or introduced. Vectors for use in the transformation of host organisms are well known in the art. Non-limiting examples of general classes of vectors include, but are not limited to, viral vectors, plasmid vectors, phage vectors, phagemid vectors, cosmid vectors, fosmid vectors, phages, artificial chromosomes, minicircles or Agrobacterium binary vectors, in double-stranded or single-stranded linear or circular form, which may or may not be self-transmissible or mobilizable. In some embodiments, viral vectors can include, but are not limited to, retroviral, lentiviral, adenoviral, adeno-associated viral, or herpes simplex viral vectors. The vectors defined herein can transform prokaryotic or eukaryotic hosts by integration into the cell genome or by existing episomally (e.g., an autonomously replicating plasmid having an origin of replication). Also included are shuttle vectors, which refer to DNA vehicles capable of (naturally or by design) replicating in two different host organisms, which host organisms can be selected from actinomycetes and related species, bacteria, and eukaryotes (e.g., higher plants, mammals, yeast, or fungal cells). In some embodiments, the nucleic acid in the vector is under the control of and operably linked to a suitable promoter or other regulatory element for transcription in the host cell. The vector can be a bifunctional expression vector that functions in a variety of hosts. In the case of genomic DNA, this can include its own promoter or other regulatory elements, and in the case of cDNA, this can be under the control of a suitable promoter or other regulatory element for expression in the host cell. Thus, the polynucleotides and nucleic acid constructs of the invention and / or expression cassettes containing them can be included in the vectors described herein and known in the art.

[0107] As used herein, "contact" and its grammatical variations refer to placing the components of a desired reaction together under conditions suitable for carrying out the desired reaction (e.g., transformation, transcriptional control, genome editing, nick generation, and / or cleavage). Thus, for example, a target nucleic acid can be contacted with a fusion protein and a guide nucleic acid of the invention to modify the target nucleic acid. In some embodiments, a target DNA can be contacted with a polynucleotide or nucleic acid construct encoding a fusion protein of the invention and a guide nucleic acid under conditions such that the fusion protein is expressed and forms a complex with the guide nucleic acid, which complex then hybridizes to the target nucleic acid to modify the target nucleic acid.

[0108] As used herein, with respect to a target nucleic acid, "modification" includes editing (e.g., mutation), covalent modification, exchange / replacement of nucleic acid / nucleobases, deletion, cleavage, nick generation, and / or transcriptional control of the target nucleic acid.

[0109] In the context of a polynucleotide of interest, "introducing" (and its grammatical variations) means presenting a nucleotide sequence of interest (e.g., a polynucleotide, a nucleic acid construct, a complex (e.g., a protein-RNA chimeric complex), and / or a guide nucleic acid) to a host organism or a cell of said organism (e.g., a host cell) in such a way that the nucleotide sequence enters the interior of the cell. Thus, for example, a polynucleotide encoding a fusion protein of the invention and a guide nucleic acid can be introduced into a cell of an organism to transform the cell.

[0110] As used herein, the term "transformation" refers to the introduction of heterologous nucleic acid into a cell. Transformation of a cell can be stable or transient. Thus, in some embodiments, a host cell or a host organism is stably transformed with a nucleotide molecule of the invention. In other embodiments, a host cell or a host organism is transiently transformed with a recombinant nucleic acid molecule of the invention.

[0111] In the context of a polynucleotide, "transient transformation" means that the polynucleotide is introduced into a cell and does not integrate into the genome of the cell.

[0112] In the context of a polynucleotide introduced into a cell, "stably introduced" means that the introduced polynucleotide is stably incorporated into the genome of the cell, and thus the cell is stably transformed with the polynucleotide.

[0113] As used herein, "stable transformation" or "stably transformed" means that a nucleic acid molecule is introduced into a cell and integrated into the genome of the cell. Thus, the integrated nucleic acid molecule can be inherited by its progeny, more particularly by progeny of multiple successive generations. As used herein, "genome" includes nuclear genome and plastid genome, and thus includes integration of nucleic acid into, for example, a chloroplast or mitochondrial genome. As used herein, "stable transformation" also refers to a transgene maintained extrachromosomally, such as as a minichromosome or a plasmid.

[0114] Transient transformation can be detected by, for example, enzyme-linked immunosorbent assay (ELISA) or Western blotting, which can detect the presence of a peptide or polypeptide encoded by one or more transgenes introduced into an organism. Stable transformation of cells can be detected by, for example, Southern blot hybridization assay of the genomic DNA of said cells, which uses a nucleic acid sequence that specifically hybridizes to the nucleotide sequence of the transgene introduced into an organism (e.g., a plant). Stable transformation of cells can be detected by, for example, Northern blot hybridization assay of the RNA of said cells, which uses a nucleic acid sequence that specifically hybridizes to the nucleotide sequence of the transgene introduced into a host organism. Stable transformation of cells can also be detected by, for example, polymerase chain reaction (PCR) or other amplification reactions (well known in the art), which employ specific primer sequences that hybridize to the target sequence of the transgene, resulting in amplification of the transgene sequence, which can be detected according to standard methods. Transformation can also be detected by direct sequencing and / or hybridization protocols well known in the art.

[0115] Thus, in some embodiments, the nucleotide sequences, nucleic acid constructs, and / or expression cassettes of the invention can be expressed transiently, and / or can be stably incorporated into the genome of a host organism. Thus, in some embodiments, the fusion proteins of the invention or polynucleotides encoding them can be introduced into cells having a guiding nucleic acid, and so no DNA remains in said cells.

[0116] The nucleic acid constructs / polynucleotides of the invention can be introduced into cells by any method known to those skilled in the art. In some embodiments of the invention, transformation of cells includes nuclear transformation. In other embodiments, transformation of cells includes plastid transformation (e.g., chloroplast transformation). In further embodiments, the nucleic acid constructs / polynucleotides of the invention can be introduced into cells via conventional breeding techniques.

[0117] Procedures for transforming both eukaryotic and prokaryotic organisms are well known and conventional in the art and are described throughout the literature (see, e.g., Jiang et al., 2013. Nat. Biotechnol. 31:233-239; Ran et al., Nature Protocols 8:2281-2308 (2013)).

[0118] Thus, nucleotide sequences can be introduced into a host organism or its cells in many ways well known in the art. The methods of the present invention are not dependent on a particular method for introducing one or more nucleotide sequences into an organism, provided that they gain entry into the interior of at least one cell of said organism. When more than one nucleotide sequence is to be introduced, they can be assembled as part of a single nucleic acid construct, or as separate nucleic acid constructs, and can be located on the same or different nucleic acid constructs. Thus, nucleotide sequences can be introduced into the target cells in a single transformation event or in separate transformation events, or alternatively, in relevant cases, nucleotide sequences can be incorporated into a plant, for example as part of a breeding protocol.

[0119] The present invention is directed to polypeptides (e.g., SEQ ID NO: 1-24), which can be used, for example, to link two or more proteins / protein domains. In some embodiments, the polypeptides of the present invention can be about 70% to 100% identical to any of the amino acid sequences of SEQ ID NO: 1-24 (e.g., 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or 100%). In some embodiments, the present invention provides polynucleotides encoding any of the amino acid sequences of SEQ ID NO: 1-24, and / or polynucleotides having 70% to 100% identity to the polynucleotides encoding any of the amino acid sequences of SEQ ID NO: 1-24 (e.g., 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or 100% identical). In some embodiments, the polynucleotides encoding any of the amino acid sequences of SEQ ID NO: 1-24 can be codon-optimized for expression in an organism.

[0120] The present invention also aims to include synthetic fusion proteins containing these polypeptides. In some embodiments, the present invention provides polypeptides that contain any one of the amino acid sequences of SEQ ID NOs: 1-24 and a polypeptide of interest. In some embodiments, the polypeptide of interest can be linked to any one of the amino acid sequences of SEQ ID NOs: 1-24 at its C-terminus and / or its N-terminus, optionally at the C-terminus and / or N-terminus. In some embodiments, the polypeptide of interest can contain two or more polypeptides of interest (e.g., 2, 3, 4, 5, 6, 7 or more), which can be the same or different, wherein at least two of the two or more polypeptides of interest can be linked to each other via any one of the amino acid sequences of SEQ ID NOs: 1-24.

[0121] Polypeptides or protein domains useful for the present invention can include, but are not limited to, polypeptides having the following activities: deaminase (deamination) activity (e.g., cytosine deaminase, adenine deaminase), nickase activity, recombinase activity, transposase activity, methylase activity, glycosylase (DNA glycosylase) activity, glycosylase inhibitor activity (e.g., uracil-DNA glycosylase inhibitor (UGI)), demethylase activity, transcriptional activation activity, transcriptional repression activity, transcriptional release factor activity, histone modification activity, nuclease activity, single-stranded RNA cleavage activity, double-stranded RNA cleavage activity, restriction endonuclease activity (e.g., Fok1), nucleic acid binding activity, methyltransferase activity, DNA repair activity, DNA damage activity, dismutase activity, alkylation activity, depurination activity, oxidation activity, pyrimidine dimer formation activity, integrase activity, transposase activity, polymerase activity, ligase activity, helicase activity, and / or photolyase activity. In some embodiments, the polypeptide of interest is adenine deaminase, cytosine deaminase, Fok1 nuclease, or uracil-DNA glycosylase inhibitor. In some embodiments, the polynucleotide of interest can be codon-optimized for expression in an organism.

[0122] In some embodiments, the polypeptide of interest is a CRISPR Cas12a polypeptide or Cas12a domain, wherein the Cas12a is linked to the C-terminus or N-terminus of any one of the amino acid sequences of SEQ ID NOs: 1-24 at its C-terminus and / or N-terminus.

[0123] In some embodiments, fusion proteins are provided that comprise Cas12a, a polypeptide of interest, and any one of the amino acid sequences of SEQ ID NOs: 1-24. In some embodiments, the amino acid sequences of SEQ ID NOs: 1-24 enable an optimal arrangement of Cas12a and one or more (e.g., 1, 2, 3, 4, 5, 6, 7 or more) polypeptides of interest (e.g., a cytosine deaminase domain, a glycosylase inhibitor (e.g., uracil-DNA glycosylase inhibitor (UGI)) relative to the Cas12a domain. The amino acid sequences of SEQ ID NOs: 1-24 can be used to link Cas12a and the polypeptide of interest in a manner that allows access to the single-stranded portion of the non-target strand for, e.g., nucleic acid modification, such as base editing.

[0124] In some embodiments, when used to link Cas12a to a polypeptide of interest, the amino acid sequences of SEQ ID NOs: 1-24 can provide different windows for modification or editing of nucleic acids. For example, the amino acid sequences of SEQ ID NOs: 1-24 that link the polypeptide of interest to Cas12a can provide a window for editing or modification of 1 to about 25 nucleotides from the corresponding PAM (protospacer adjacent motif) in the target nucleic acid (e.g., DNA) (e.g., an editing / modification window of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 nucleotides from the PAM and any range or value therein). In some embodiments, the editing or modification window can be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 to about 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 nucleotides from the PAM (e.g., 1 to 20, 1 to 15, 1 to 10, 3 to 15, 4 to 10, 5 to 25, 5 to 20, 5 to 15, 5 to 10, 7 to 15 nucleotides, etc. from the PAM).

[0125] Cas12a is a type V clustered regularly interspaced short palindromic repeat (CRISPR)-Cas nuclease. Cas12a differs from the better-known type II CRISPR Cas9 nuclease in several respects. For example, Cas9 recognizes a G-rich protospacer adjacent motif (PAM) (3'-NGG) at the 3' of its guide RNA (gRNA, sgRNA) binding site (protospacer, target nucleic acid, target DNA), while Cas12a recognizes a T-rich PAM (5'-ttN, 5'-TTTN) located at the 5' of the target nucleic acid. In fact, the orientations by which Cas9 and Cas12a bind their guide RNAs are nearly opposite with respect to their N and C termini. Further, the Cas12a enzyme uses a single guide RNA (gRNA, CRISPR array, crRNA), rather than the dual guide RNAs (sgRNA (e.g., crRNA and tracrRNA)) found in the native Cas9 system, and Cas12a processes its own gRNA. Additionally, Cas12a nuclease activity generates staggered DNA double-strand breaks, rather than the blunt ends generated by Cas9 nuclease activity, and Cas12a relies on a single RuvC domain to cleave both DNA strands, while Cas9 utilizes an HNH domain and an RuvC domain for cleavage.

[0126] The CRISPR Cas12a polypeptides or CRISPR Cas12a domains useful for the present invention can be any known or later identified Cas12a nuclease (formerly known as Cpf1) (see, e.g., U.S. Patent No. 9,790,490, which is incorporated by reference for its disclosure regarding Cpf1 (Cas12a) sequences). The terms "Cas12a", "Cas12a polypeptide", or "Cas12a domain" refer to an RNA-guided nuclease comprising a Cas12a polypeptide or a fragment thereof that comprises the guide nucleic acid binding domain of Cas12a, and / or the active, inactive, or partially active DNA cleavage domain of Cas12a. In some embodiments, the Cas12a useful for the present invention can comprise a mutation in the nuclease active site (e.g., the RuvC site of the Cas12a domain). A Cas12a domain or Cas12a polypeptide that has a mutation in its nuclease active site and thus no longer comprises nuclease activity is generally referred to as deadCas12a (e.g., dCas12a). In some embodiments, a Cas12a domain or Cas12a polypeptide that has a mutation in its nuclease active site can have impaired activity.

[0127] In some embodiments, the Cas12a domain can include, but is not limited to, any of the amino acid sequences of SEQ ID NOs: 29-45 (e.g., SEQ ID NO: 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 45); or a polynucleotide encoding the same. In some embodiments, the fusion protein of the present invention can comprise a Cas12a domain from the bacterium Lachnospiraceae bacterium ND2006 Cas12a (LbCas12a) (e.g., SEQ ID NO: 29).

[0128] In some embodiments, the polynucleotide encoding the Cas12a domain can be codon-optimized for expression in an organism. Thus, in some embodiments, the present invention provides a polynucleotide having at least about 70% identity (e.g., about 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% identity) to a polynucleotide encoding an amino acid sequence of any of SEQ ID NOs: 29-45.

[0129] In some embodiments, a type V clustered regularly interspaced short palindromic repeat (CRISPR)-associated (Cas) (CRISPR-Cas) system is provided, the system comprising: (a) a fusion protein comprising a Cas12a domain, a linker comprising any of the amino acid sequences of SEQ ID NOs: 1-24, and a polypeptide of interest, or a nucleic acid encoding the fusion protein, wherein the Cas12a domain is linked to the polypeptide of interest via any of the amino acid sequences of SEQ ID NOs: 1-24; and (b) a guide nucleic acid (CRISPR RNA, CRISPR DNA, crRNA, crDNA) comprising a spacer sequence and a repeat sequence, wherein the guide nucleic acid is capable of forming a complex with the Cas12a domain of the fusion protein, and the spacer sequence is capable of hybridizing to a target nucleic acid, thereby guiding the Cas12a domain and the polypeptide of interest to the target nucleic acid, whereby the system is capable of modifying (e.g., cleaving or editing) or regulating (e.g., regulating transcription) the target nucleic acid.

[0130] In some embodiments, a fusion protein is provided, which comprises Cas12a, a polypeptide of interest, and any of the amino acid sequences of SEQ ID NOs: 1-24, wherein the polypeptide of interest is a cytosine deaminase polypeptide or domain.

[0131] In some embodiments, the present invention provides a fusion protein comprising: (a) a Cas12a domain that specifically binds to a target nucleic acid sequence when associated with a bound guide nucleic acid (e.g., gRNA); (b) a cytidine deaminase domain that deaminates cytosine bases in the single-stranded portion of the target nucleic acid sequence when associated with the Cas12a domain and the gRNA; and (c) a uracil glycosylase inhibitor (UGI) domain that inhibits uracil-DNA glycosylase, wherein the Cas12a domain is linked to the cytidine deaminase domain or the UGI domain via any one of the amino acid sequences of SEQ ID NOs: 1-24. In some embodiments, the N-terminus of the Cas12a domain can be linked to the C-terminus of the cytidine deaminase domain via any one of the amino acid sequences of SEQ ID NOs: 1-5, the C-terminus of the Cas12a domain can be linked to the N-terminus of the UGI domain via any one of the amino acid sequences of SEQ ID NOs: 6-12, the N-terminus of the cytidine deaminase domain can be linked to the C-terminus of the UGI domain via any one of the amino acid sequences of SEQ ID NOs: 13-16, the N-terminus of the Cas12a domain can be linked to the C-terminus of the UGI domain via any one of the amino acid sequences of SEQ ID NOs: 17-19, and / or the N-terminus of the cytidine deaminase domain can be linked to the C-terminus of the Cas12a domain via any one of the amino acid sequences of SEQ ID NOs: 20-24. In some embodiments, when the N-terminus of the Cas12a domain is linked to the C-terminus of the cytidine deaminase domain via any one of the amino acid sequences of SEQ ID NOs: 1-5, the C-terminus of the Cas12a domain can be linked to the UGI domain via a GS linker. In some embodiments, when the C-terminus of the Cas12a domain is linked to the N-terminus of the UGI domain via any one of the amino acid sequences of SEQ ID NOs: 6-12, the N-terminus of the Cas12a domain can be linked to the cytidine deaminase domain via a GS linker. In some embodiments, when the N-terminus of the Cas12a domain is linked to the C-terminus of the UGI domain via any one of the amino acid sequences of SEQ ID NOs: 17-19, the C-terminus of the Cas12a can be linked to the cytidine deaminase via a GS linker.In some embodiments, when the N-terminus of the cytosine deaminase domain is linked to the C-terminus of the Cas12a domain via any one of the amino acid sequences of SEQ ID NOs: 20-24, the C-terminus of the Cas12a is linked to the cytosine deaminase via a GS linker. Exemplary fusion proteins of the invention include, but are not limited to, the amino acid sequences of SEQ ID NOs: 49-72.

[0132] In some embodiments, there is provided a fusion protein comprising: (a) a cytosine deaminase domain; (b) a Cas12a domain; and (c) a uracil-DNA glycosylase inhibitor (UGI) domain, wherein the C-terminus of the cytosine deaminase domain is linked to the N-terminus of the Cas12a domain via any one of the amino acid sequences of SEQ ID NOs: 1-5, and the C-terminus of the Cas12a domain is linked to the N-terminus of the UGI domain, or the C-terminus of the Cas12a domain is linked to the N-terminus of the UGI domain via any one of the amino acid sequences of SEQ ID NOs: 6-9, and the C-terminus of the cytosine deaminase domain is linked to the N-terminus of the Cas12a domain. In some embodiments, the C-terminus of the Cas12a domain may be linked to the N-terminus of the UGI domain via a GS linker. In some embodiments, the C-terminus of the cytosine deaminase domain is linked to the N-terminus of the Cas12a domain via a GS linker. In some embodiments, the C-terminus of the cytosine deaminase domain is linked to the N-terminus of the Cas12a domain via the amino acid sequence of SEQ ID NO: 29. Exemplary fusion proteins of the invention include, but are not limited to, any one of the amino acid sequences of SEQ ID NOs: 64-72.

[0133] In some embodiments, there is provided a fusion protein comprising: (a) a Cas12a (Cpf1) domain; (b) a uracil-DNA glycosylase inhibitor (UGI) domain; and (c) a cytosine deaminase domain, wherein the C-terminus of the Cas12a domain is linked to the N-terminus of the UGI domain via any one of the amino acid sequences of SEQ ID NOs: 10-12, and the C-terminus of the UGI domain is linked to the N-terminus of the cytosine deaminase domain via any one of the amino acid sequences of SEQ ID NOs: 13-16, wherein the amino acid sequences of SEQ ID NOs: 10-12 and SEQ ID NOs: 13-16 are independently selected. Exemplary fusion proteins of the invention include, but are not limited to, any one of the amino acid sequences of SEQ ID NOs: 58-63.

[0134] In some embodiments, a fusion protein is provided that comprises: (a) a uracil-DNA glycosylase inhibitor (UGI) domain; (b) a Cas12a (Cpf1) domain, wherein the Cas12a domain comprises a mutation at the nuclease active site; and (c) a cytosine deaminase domain, wherein the C-terminus of the UGI domain is linked to the N-terminus of the Cas12a domain via any one of the amino acid sequences of SEQ ID NOs: 17-19, and the C-terminus of the Cas12a domain is linked to the N-terminus of the cytosine deaminase domain, or wherein the C-terminus of the UGI domain is linked to the N-terminus of the Cas12a domain, and the C-terminus of the Cas12a domain is linked to the N-terminus of the cytosine deaminase domain via any one of the amino acid sequences of SEQ ID NOs: 20-24. In some embodiments, the C-terminus of the Cas12a domain is linked to the N-terminus of the cytosine deaminase domain via a GS linker. In some embodiments, the C-terminus of the Cas12a domain is linked to the N-terminus of the cytosine deaminase domain via the amino acid sequence of SEQ ID NO: 28. In some embodiments, the C-terminus of the UGI domain is linked to the N-terminus of the Cas12a domain via a GS linker. Exemplary fusion proteins of the invention include, but are not limited to, any one of the amino acid sequences of SEQ ID NOs: 49-72.

[0135] The cytosine deaminase (or cytidine deaminase) useful for the present invention can be any known or later identified cytosine deaminase from any organism (see, for example, U.S. Patent No. 10,167,457; and Thuronyi et al., Nat. Biotechnol. 37:1070-1079 (2019), each incorporated herein by reference for its disclosure regarding cytosine deaminase). The cytosine deaminase can catalyze the hydrolysis and deamination of cytidine or deoxycytidine to uridine or deoxyuridine, respectively. In some embodiments, the deaminase polypeptide or deaminase domain is a cytidine deaminase domain that catalyzes the hydrolysis and deamination of cytosine to uracil. In some embodiments, the cytosine deaminase can be a variant of a naturally occurring cytosine deaminase, including but not limited to primates (e.g., human, monkey, chimpanzee, gorilla), dog, cow, rat, or mouse. Thus, in some embodiments, the cytosine deaminase useful for the present invention can be about 70% to 100% identical to the wild-type cytosine deaminase (e.g., about 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a naturally occurring cytosine deaminase, and any range or value therein). In some embodiments, the polynucleotide encoding the cytosine deaminase polypeptide / domain can be codon-optimized for expression in an organism.

[0136] In some embodiments, the cytosine deaminase useful for the present invention can be an apolipoprotein B mRNA editing complex (APOBEC) family deaminase. In some embodiments, the cytosine deaminase can be APOBEC1 deaminase, APOBEC2 deaminase, APOBEC3A deaminase, APOBEC3B deaminase, APOBEC3C deaminase, APOBEC3D deaminase, APOBEC3F deaminase, APOBEC3G deaminase, APOBEC3H deaminase, APOBEC4 deaminase, human activation-induced deaminase (hAID), rAPOBEC1, FERNY, and / or CDA1, optionally pmCDA1, atCDA1 (e.g., At2g19570), and / or an evolved version thereof. In some embodiments, the cytosine deaminase can be APOBEC1 deaminase having the amino acid sequence of SEQ ID NO: 46. In some embodiments, the cytosine deaminase can be APOBEC3A deaminase having the amino acid sequence of SEQ ID NO: 47. In some embodiments, the cytosine deaminase useful for the present invention can be about 70% to 100% identical to the amino acid sequence of a naturally occurring cytosine deaminase (e.g., 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 100% identical). In some embodiments, the cytosine deaminase useful for the present invention can be about 70% to 99.5% identical to the amino acid sequence of SEQ ID NO: 46 or SEQ ID NO: 47 (e.g., 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5% identical) (e.g., at least 80%, at least 85%, at least 90%, at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5% identical to the amino acid sequence of SEQ ID NO: 46 or SEQ ID NO: 47). In some embodiments, the polynucleotide encoding the cytosine deaminase can be codon-optimized for expression in an organism, and the codon-optimized polypeptide can be about 70% to 99.5% identical to the reference polynucleotide.

[0137] For the present invention, a "uracil glycosylase inhibitor" useful for the present invention can be any protein that inhibits uracil-DNA glycosylase base excision repair enzyme. In some embodiments, the UGI domain comprises wild-type UGI or a fragment thereof. In some embodiments, the UGI domain useful for the present invention can be about 70% to 100% identical to the amino acid sequence of a naturally occurring UGI domain (e.g., 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or 100% identical, and any range or value therein). In some embodiments, the UGI domain can comprise the amino acid sequence of SEQ ID NO: 48, or a polypeptide having about 70% to 99.5% identity to the amino acid sequence of SEQ ID NO: 48 (e.g., at least 80%, at least 85%, at least 90%, at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or at least 99.5% identical to the amino acid sequence of SEQ ID NO: 48). For example, in some embodiments, the UGI domain can comprise a fragment of the amino acid sequence of SEQ ID NO: 48 that is 100% identical to a portion of the amino acid sequence of SEQ ID NO: 48 (e.g., 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80 consecutive nucleotides; e.g., about 10, 15, 20, 25, 30, 35, 40, 45 to about 50, 55, 60, 65, 70, 75, 80 consecutive nucleotides). In some embodiments, the UGI domain can be a variant of a known UGI (e.g., SEQ ID NO: 48) that has 70% to 99.5% identity to the known UGI (e.g., 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% identity, and any range or value therein). In some embodiments, the polynucleotide encoding UGI can be codon-optimized for expression in an organism, and the codon-optimized polynucleotide can be about 70% to 99.5% identical to a reference polynucleotide.

[0138] The fusion proteins of the invention described herein that comprise a Cas12a domain linked to a polypeptide of interest can be used in combination with a guide RNA (gRNA, CRISPR array, CRISPR RNA, crRNA) designed to function with the Cas12a domain to modify a target nucleic acid. Guide nucleic acids (CRISPR RNA, CRISPR DNA, crRNA, crDNA) useful for the invention comprise a spacer sequence and a repeat sequence. The guide nucleic acid is capable of forming a complex with the Cas12a domain of the fusion protein, and the spacer sequence is capable of hybridizing to the target nucleic acid, thereby guiding the Cas12a domain and the polypeptide of interest to the target nucleic acid, wherein the target nucleic acid is modified (e.g., cleaved or edited) or regulated (e.g., transcriptional regulation) by the polypeptide of interest of the fusion protein. As an example, the fusion proteins described herein that comprise a Cas12a domain linked to a cytidine deaminase domain can be used in combination with a Cas12a guide nucleic acid to modify a target nucleic acid, wherein the cytidine deaminase domain of the fusion protein deaminates a cytosine base in the target nucleic acid, thereby editing the target nucleic acid.

[0139] As used herein, "guide nucleic acid", "guide RNA", "gRNA", "CRISPR RNA / DNA", "crRNA", or "crDNA" refers to a nucleic acid that comprises at least one spacer sequence that is complementary to (and hybridizes to) a target DNA (e.g., a protospacer), and at least one repeat sequence (e.g., a repeat sequence of a type V Cas12a CRISPR-Cas system, or a fragment or portion thereof), wherein the repeat sequence is linked to the 5' end of the spacer sequence. The design of the gRNA of the invention is based on the type V Cas12a CRISPR-Cas system. In some embodiments, the gRNA for Cas12a can comprise (from 5' to 3'): a repeat sequence (full length or a portion thereof ("stem"); e.g., a pseudoknot-like structure); and a spacer sequence. In some embodiments, the guide nucleic acid can comprise more than one repeat sequence-spacer sequence (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10 or more repeat sequence-spacer sequences) (e.g., repeat-spacer-repeat; e.g., repeat-spacer-repeat-spacer-repeat-spacer-repeat-spacer-repeat-spacer-repeat-spacer, etc.). The guide nucleic acids of the invention are synthetic, artificial and not found in nature. The gRNA can be quite long and can be used as an aptamer (e.g., in an MS2 recruitment strategy) or other RNA structures hanging off the spacer.

[0140] As used herein, "repeat sequence" refers to, for example, any repeat sequence of the wild-type CRISPR Cas12a locus or the repeat sequence of a synthetic crRNA. Repeat sequences useful for the present invention can be any known or later identified repeat sequence of a CRISPR Cas12a locus (type V), or it can be a synthetic repeat sequence designed to function in a type V CRISPR-Cas system. The repeat sequence can include a hairpin structure and / or a stem-loop structure. In some embodiments, the repeat sequence can form a pseudoknot-like structure (i.e., a "handle") at its 5' end. Thus, in some embodiments, the repeat sequence can be identical or substantially identical (e.g., at least 70% identical) to a repeat sequence from a wild-type type V CRISPR locus. Repeat sequences from wild-type Cas12a (type V) CRISPR loci can be determined by established algorithms, such as using CRISPRfinder provided by CRISPRdb (see Grissa et al., Nucleic Acids Res. 35 (Web Server issue): W52-7). In some embodiments, the repeat sequence or a portion thereof is linked to the 5' end of the spacer sequence to form a repeat-spacer sequence (e.g., guide RNA, crRNA).

[0141] In some embodiments, the repeat sequence comprises at least 10 nucleotides, consists essentially of at least 10 nucleotides, or consists of at least 10 nucleotides, depending on the particular repeat sequence and whether the guide RNA containing the repeat sequence is processed or unprocessed (e.g., about 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50 to 100 or more nucleotides, or any range or value therein). In some embodiments, the repeat sequence comprises, consists essentially of, or consists of the following nucleotides: about 10 to about 20, about 10 to about 30, about 10 to about 45, about 10 to about 50, about 15 to about 30, about 15 to about 40, about 15 to about 45, about 15 to about 50, about 20 to about 30, about 20 to about 40, about 20 to about 50, about 30 to about 40, about 40 to about 80, about 50 to about 100 or more nucleotides.

[0142] The repeat sequence linked to the 5' end of the spacer sequence can include a portion of the repeat sequence (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 or more contiguous nucleotides of the wild-type repeat sequence). In some embodiments, the length of the portion of the repeat sequence linked to the 5' end of the spacer sequence can be from about five to about ten consecutive nucleotides (e.g., about 5, 6, 7, 8, 9, 10 nucleotides), and has at least 90% identity (e.g., at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more) with the same region (e.g., the 5' end) of the wild-type Cas12a repeat nucleotide sequence. In some embodiments, the portion of the repeat sequence contains a pseudoknot-like structure (e.g., a "stem") at its 5' end.

[0143] As used herein, an "spacer sequence" is a nucleotide sequence that is complementary to a target nucleic acid (e.g., target DNA) (e.g., protospacer). The spacer sequence can be fully complementary or substantially complementary to the target nucleic acid (e.g., at least about 70% complementary (e.g., about 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more)). Thus, in some embodiments, the spacer sequence can have one, two, three, four or five mismatches compared to the target nucleic acid, and the mismatches can be contiguous or non - contiguous. In some embodiments, the spacer sequence can have 70% complementarity to the target nucleic acid. In other embodiments, the spacer nucleotide sequence can have 80% complementarity to the target nucleic acid. In still other embodiments, the spacer nucleotide sequence can have 85%, 90%, 95%, 96%, 97%, 98%, 99% or 99.5% complementarity to the target nucleic acid (protospacer), etc. In some embodiments, the spacer sequence is 100% complementary to the target nucleic acid. The spacer sequence can have a length of from about 15 nucleotides to about 30 nucleotides (e.g., 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 nucleotides, or any range or value therein). Thus, in some embodiments, the spacer sequence can have complete or substantial complementarity over a region of a target nucleic acid (e.g., protospacer) that is at least about 15 nucleotides to about 30 nucleotides in length. In some embodiments, the length of the spacer is about 20 nucleotides. In some embodiments, the length of the spacer is about 23 nucleotides.

[0144] In some embodiments, the 5' region of the spacer sequence of the guide RNA can be identical to the target DNA, while the 3' region of the spacer can be substantially identical to the target DNA, and thus the overall complementarity of the spacer sequence to the target DNA can be less than 100%. Thus, for example, the first 1, 2, 3, 4, 5, 6, 7, 8, etc. nucleotides (i.e., the seed region) in the 5' region of a spacer sequence of, for example, 20 nucleotides can be 100% complementary to the target DNA, while the remaining nucleotides in the 3' region of the spacer sequence are substantially complementary to the target DNA (e.g., at least about 70% complementary). In some embodiments, the first 1 to 8 nucleotides (e.g., the first 1, 2, 3, 4, 5, 6, 7, 8 nucleotides, and any range therein) at the 5' end of the spacer sequence can be 100% complementary to the target DNA, while the remaining nucleotides in the 3' region of the spacer sequence are substantially complementary to the target DNA (e.g., at least about 50% complementary (e.g., 50%, 55%, 60%, 65%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more)). In some embodiments, the length of the seed region of the spacer can be about 5 to 6 nucleotides. In some embodiments, the length of the seed region of the spacer is 5 nucleotides. In some embodiments, the length of the seed region of the spacer is 6 nucleotides.

[0145] As used herein, "target nucleic acid", "target DNA", "target nucleotide sequence", "target region", or "target region in the genome" refers to a region of the genome of an organism that is perfectly complementary (100% complementary) or substantially complementary (e.g., at least 70% complementary (e.g., 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more)) to the spacer sequence in the guide RNA of the invention. A target region useful for the CRISPR-Cas12a system is immediately located 3' of a PAM sequence in the genome of an organism. The target region can be selected from any at least 15 consecutive nucleotides (e.g., 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 nucleotides, etc.) immediately adjacent to the PAM sequence.

[0146] "The protospacer sequence" refers to the target double-stranded DNA, and particularly to that portion of the target DNA (e.g., the target region in the genome) that is fully or substantially complementary (and hybridizes) to the spacer sequence of a CRISPR repeat-spacer sequence (e.g., guide RNA, CRISPR array, crRNA). In the case of the type V CRISPR-Cas Cas12a system, the protospacer sequence is flanked (immediately adjacent) by a protospacer adjacent motif (PAM). The PAM is located at the 5' end on the non-target strand and the 3' end on the target strand (as an example, see below).

[0147]

[0148] The canonical Cas12a PAM is T-rich. In some embodiments, the canonical Cas12a PAM sequence can be 5'-TTN, 5'-TTTN or 5'-TTTV. In some embodiments, non-canonical PAMs can be used, but they may be less efficient.

[0149] Additional PAM sequences can be determined by those skilled in the art through established experimental and computational methods. Thus, for example, experimental methods include targeting sequences flanked by all possible nucleotide sequences and identifying sequence members that do not undergo targeting, e.g., by transformation of target plasmid DNA (Esvelt et al., 2013. Nat. Methods 10:1116-1121; Jiang et al., 2013. Nat. Biotechnol. 31:233-239). In some aspects, computational methods can include performing a BLAST search of native spacers to identify the original target DNA sequences in phages or plasmids and aligning these sequences to determine the conserved sequences adjacent to the target sequences (Briner and Barrangou, 2014. Appl. Environ. Microbiol. 80:994-1001; Mojica et al., 2009. Microbiology 155:733-740).

[0150] In some embodiments, a complex or composition is provided that comprises one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8 or more) fusion proteins of the invention and one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8 or more) guide nucleic acids (e.g., CRISPR RNA / DNA, e.g., crRNA / crDNA). In some embodiments, a polynucleotide or nucleic acid construct is provided that encodes a polypeptide, fusion protein, guide nucleic acid, and / or complex of the invention. In some embodiments, a nucleic acid construct, expression cassette, and / or vector is provided that comprises a polynucleotide of the invention and / or one or more guide nucleic acids. In some embodiments, the polynucleotide encoding the fusion protein of the invention can be encoded on the same or a separate polynucleotide, nucleic acid construct, expression cassette, or vector as the polynucleotide comprising the guide nucleic acid. When the fusion protein is encoded on a polynucleotide, nucleic acid construct, expression cassette, or vector separate from the polynucleotide comprising the guide nucleic acid, the polynucleotide, nucleic acid construct, expression cassette, or vector encoding the fusion protein of the invention (e.g., contacting a target nucleic acid) can be provided before, simultaneously with, or after the guide nucleic acid (e.g., contacting the target nucleic acid) is provided.

[0151] In some embodiments, the polynucleotides, nucleic acid constructs, expression cassettes, and / or vectors of the invention can be codon-optimized for expression in an organism. In some embodiments, the optimized polynucleotides, nucleic acid constructs, or expression cassettes of the invention can be about 70% to 100% identical (e.g., about 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or 100%) to the polynucleotides, nucleic acid constructs, or expression cassettes encoding the polypeptides, fusion proteins, and complexes of the invention.

[0152] In some embodiments, a cell is provided that comprises one or more polynucleotides, guide nucleic acids, nucleic acid constructs, expression cassettes, or vectors of the invention.

[0153] The polypeptides, fusion proteins, guide RNAs, complexes, and compositions of the invention, and the polynucleotides / nucleic acid constructs / expression cassettes / vectors encoding them can be used to modify a target nucleic acid and / or its expression.

[0154] In some embodiments, the fusion protein of the invention is a cytosine base editor (ABE) for use in base editing of a target nucleic acid, wherein the fusion protein comprises a Cas12a domain linked to a cytosine deaminase domain.

[0155] In some embodiments, methods of modifying a target nucleic acid are provided, the methods comprising contacting the target nucleic acid with: (a)(i) a fusion protein of the invention, and (a)(ii) a guide nucleic acid (e.g., CRISPR RNA, CRISPR DNA, crRNA, crDNA); (b) a complex comprising the fusion protein of the invention and a guide nucleic acid; (c) a composition comprising the fusion protein of the invention and a guide nucleic acid; and / or (d) a system of the invention, thereby modifying the target nucleic acid. The target nucleic acid can be contacted with the fusion protein before, simultaneously with, or after contacting the target nucleic acid with the guide nucleic acid.

[0156] In some embodiments, methods of modifying a target nucleic acid are provided, the methods comprising contacting the target nucleic acid with a fusion protein comprising any one of the amino acid sequences of SEQ ID NOs: 49-72 and a guide nucleic acid. The target nucleic acid can be contacted with the fusion protein of the invention before, simultaneously with, or after contacting the target nucleic acid with the guide nucleic acid.

[0157] In some embodiments, methods of modifying a target nucleic acid are provided, the methods comprising contacting a cell or cell-free system comprising the target nucleic acid, under conditions, with: (a)(i) a polynucleotide encoding a polypeptide or a fusion protein of the invention, or an expression cassette or vector comprising the same, and (a)(ii) a guide nucleic acid, and / or an expression cassette or vector comprising the same; and / or (b) a nucleic acid construct encoding a complex comprising the fusion protein of the invention and a guide nucleic acid, and / or an expression cassette or vector comprising the same, thereby modifying the target nucleic acid, the conditions being such that the fusion protein is expressed and forms a complex with the guide nucleic acid, the complex hybridizing to the target nucleic acid. When provided on separate constructs, the target nucleic acid can be contacted with the polynucleotide encoding the fusion protein, nucleic acid construct, expression cassette, or vector before, simultaneously with, or after contacting the target nucleic acid with the guide nucleic acid.

[0158] In some embodiments, methods of modifying a target nucleic acid are provided, the methods comprising contacting a cell or cell-free system comprising the target nucleic acid with: a polynucleotide encoding a fusion protein comprising any one of the amino acid sequences of SEQ ID NOs: 50-78, or an expression cassette or vector comprising the same; and a guide nucleic acid, or an expression cassette or vector comprising the same, such that the target nucleic acid is modified, the conditions being such that the fusion protein is expressed and forms a complex with the guide nucleic acid, the complex hybridizing to the target nucleic acid. When provided on separate constructs, the target nucleic acid can be contacted with the polynucleotide, nucleic acid construct, expression cassette or vector encoding the fusion protein before, simultaneously with or after contacting the target nucleic acid with the guide nucleic acid.

[0159] In some embodiments, the present invention provides methods of editing a target nucleic acid, the methods comprising contacting the target nucleic acid with: (a)(i) a fusion protein of the present invention, and (a)(ii) a guide nucleic acid; (b) a complex comprising a fusion protein of the present invention and a guide nucleic acid; (c) a composition comprising (i) a fusion protein of the present invention and (ii) a guide nucleic acid; and / or (d)(i) a CRISPR-Cas system of the present invention, wherein the cytosine deaminase domain converts cytosine (C) in the target nucleic acid to thymine (T), thereby editing the target nucleic acid to produce a (point) mutation. The target nucleic acid can be contacted with the fusion protein of the present invention before, simultaneously with or after contacting the target nucleic acid with the guide nucleic acid.

[0160] In some embodiments, methods of editing a target nucleic acid are provided, the methods comprising contacting the target nucleic acid with a fusion protein comprising any one of the amino acid sequences of SEQ ID NOs: 49-72 and a guide nucleic acid, thereby editing the target nucleic acid. The target nucleic acid can be contacted with the fusion protein of the present invention before, simultaneously with or after contacting the target nucleic acid with the guide nucleic acid.

[0161] In some embodiments, methods of editing a target nucleic acid are provided, the methods comprising contacting a cell or cell-free system comprising the target nucleic acid, under conditions, with: (a)(i) a polynucleotide encoding a fusion protein of the invention, and / or an expression cassette or vector comprising the same, and (a)(ii) a guide nucleic acid, and / or an expression cassette or vector comprising (a)(i) and / or (a)(ii); and / or (b) a nucleic acid construct encoding a complex of a fusion protein of the invention and a guide nucleic acid, or an expression cassette or vector comprising the same, the conditions being such that the fusion protein is expressed and forms a complex with the guide nucleic acid, the complex hybridizes to the target nucleic acid, wherein the cytosine deaminase domain converts cytosine (C) in the target nucleic acid to thymine (T), thereby editing the target nucleic acid to create a (point) mutation. When provided on separate constructs, the target nucleic acid can be contacted with the fusion protein before, simultaneously with, or after contacting the target nucleic acid with the guide nucleic acid.

[0162] In some embodiments, methods of editing a target nucleic acid are provided, the methods comprising contacting a cell or cell-free system comprising the target nucleic acid, under conditions, with: a polynucleotide encoding a fusion protein comprising any one of the amino acid sequences of SEQ ID NOs: 49-72, or an expression cassette or vector comprising the same; and a guide nucleic acid, or an expression cassette or vector comprising the same, thereby editing the target nucleic acid, the conditions being such that the fusion protein is expressed and forms a complex with the guide nucleic acid, the complex hybridizes to the target nucleic acid. The polynucleotide encoding a fusion protein comprising any one of the amino acid sequences of SEQ ID NOs: 49-72 can be present on the same expression cassette or vector comprising the guide nucleic acid. When the polynucleotide encoding a fusion protein comprising any one of the amino acid sequences of SEQ ID NOs: 49-72 is on an expression cassette or vector separate from the polynucleotide comprising the guide nucleic acid, the target nucleic acid can be contacted with the expression cassette / vector comprising the fusion protein before, simultaneously with, or after contacting the target nucleic acid with the expression cassette / vector comprising the guide nucleic acid.

[0163] In some embodiments, the present invention provides methods of editing target domains / polypeptides useful for base editing that can be used with the present invention. As used herein, "cytosine deaminase" and "cytidine deaminase" refer to a polypeptide or domain thereof that catalyzes or is capable of catalyzing the deamination of cytosine, as the polypeptide or domain catalyzes or is capable of catalyzing the removal of an amino group from a cytosine base. Thus, a cytosine deaminase can cause cytosine to be converted to thymidine (via a uracil intermediate), thereby causing a C to T transition in the genome, or a G to A transition (in the complementary strand). Thus, in some embodiments, the cytosine deaminase encoded by the polynucleotides of the present invention generates a C→T transition in the sense (e.g., "+", template) strand of the target nucleic acid, or a G→A transition in the antisense (e.g., "-", complementary) strand of the target nucleic acid. In some embodiments, the cytosine deaminase encoded by the polynucleotides of the present invention generates a C to T or G to A transition (in the complementary strand) in the genome.

[0164] Cytosine deaminases useful for the present invention can be any known or later identified cytosine deaminase from any organism (see, e.g., U.S. Patent No. 10,167,457; and Thuronyi et al., Nat. Biotechnol. 37:1070-1079 (2019), each incorporated herein by reference for its disclosure regarding cytosine deaminases). A cytosine deaminase can catalyze the hydrolytic deamination of cytidine or deoxycytidine to uridine or deoxyuridine, respectively. Thus, in some embodiments, a deaminase or deaminase domain useful for the present invention can be a cytidine deaminase domain that catalyzes the hydrolytic deamination of cytosine to uracil. In some embodiments, a cytosine deaminase can be a variant of a naturally occurring cytosine deaminase, including but not limited to primates (e.g., human, monkey, chimpanzee, gorilla), dog, cow, rat, or mouse. Thus, in some embodiments, a cytosine deaminase useful for the present invention can be about 70% to about 100% identical to a wild-type cytosine deaminase (e.g., about 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a naturally occurring cytosine deaminase, and any range or value therein).

[0165] In some embodiments, the cytosine deaminase useful for the present invention can be a member of the apolipoprotein B mRNA editing complex (APOBEC) family of deaminases. In some embodiments, the cytosine deaminase can be APOBEC1 deaminase, APOBEC2 deaminase, APOBEC3A deaminase, APOBEC3B deaminase, APOBEC3C deaminase, APOBEC3D deaminase, APOBEC3F deaminase, APOBEC3G deaminase, APOBEC3H deaminase, APOBEC4 deaminase, human activation-induced deaminase (hAID), rAPOBEC1, FERNY, and / or CDA1, optionally pmCDA1, atCDA1 (e.g., At2g19570) and its evolved versions. In some embodiments, the cytosine deaminase can be APOBEC1 deaminase, which optionally has the amino acid sequence of SEQ ID NO:46 or SEQ ID NO:79. In some embodiments, the cytosine deaminase can be APOBEC3A deaminase, which optionally has the amino acid sequence of SEQ ID NO:47. In some embodiments, the cytosine deaminase can be CDA1 deaminase, optionally CDA1 having the amino acid sequence of SEQ ID NO:76. In some embodiments, the cytosine deaminase can be FERNY deaminase, optionally FERNY having the amino acid sequence of SEQ ID NO:77 or SEQ ID NO:80. In some embodiments, the cytosine deaminase can be hAID deaminase, optionally hAID having the amino acid sequence of SEQ ID NO:81 or SEQ ID NO:82. In some embodiments, the cytosine deaminase useful for the present invention can be about 70% to about 100% identical (e.g., 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or 100% identical) to the amino acid sequence of a naturally occurring cytosine deaminase (e.g., an "evolved deaminase") (see, e.g., SEQ ID NO:78, SEQ ID NO:79, SEQ ID NO:80, SEQ ID NO:82).In some embodiments, a cytosine deaminase useful for the present invention can be about 70% to about 99.5% identical to the amino acid sequence of SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:76, SEQ ID NO:77, SEQ ID NO:78, SEQ ID NO:79, SEQ ID NO:80, SEQ ID NO:81, or SEQ ID NO:82 (e.g., about 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5% identical) (e.g., at least 80%, at least 85%, at least 90%, at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5% identical to the amino acid sequence of SEQ ID NO:46, SEQ ID NO:47, SEQ ID NO:76, SEQ ID NO:77, SEQ ID NO:78, SEQ ID NO:79, SEQ ID NO:80, SEQ ID NO:81, or SEQ ID NO:82). In some embodiments, a polynucleotide encoding a cytosine deaminase can be codon-optimized for expression in plants, and the codon-optimized polypeptide can be about 70% to 99.5% identical to a reference polynucleotide.

[0166] The fusion proteins of the present invention, polynucleotides encoding them, and nucleic acid constructs can be used in combination with a guide nucleic acid to modify a target nucleic acid, including but not limited to generating C→T or G→A mutations in the target nucleic acid (including but not limited to, plasmid sequences); generating C→T or G→A mutations in a coding sequence to alter amino acid identity; generating C→T or G→A mutations in a coding sequence to generate a stop codon; generating C→T or G→A mutations in a coding sequence to disrupt a start codon; generating point mutations in genomic DNA to disrupt transcription factor binding; generating point mutations in genomic DNA to disrupt splice junctions; and / or other nucleic acid modifications generated by a fusion protein comprising a Cas12a domain fused to another domain (a polypeptide of interest) via any one of the amino acid sequences SEQ ID NO:1-24 (e.g., a peptide linker).

[0167] The fusion proteins of the present invention, polynucleotides encoding them, and nucleic acid constructs can be useful for modifying the target nucleic acids of any organism, including but not limited to animals, plants, fungi, archaea, or bacteria. Animals can include but are not limited to, mammals, insects, fish, birds, etc.

[0168] Exemplary mammals for which the present invention can be useful include, but are not limited to, primates (human and non-human (e.g., chimpanzee, baboon, monkey, gorilla, etc.)), cats, dogs, mice, rats, ferrets, gerbils, hamsters, cows, pigs, horses, goats, donkeys, or sheep.

[0169] The fusion proteins of the present invention and the polynucleotides and nucleic acid constructs encoding them can be used to modify the target nucleic acids of any plant or plant part. Any plant (or group of plants grouped, for example, by genus or higher taxonomic order) can be employed in practicing the present invention, including angiosperms, gymnosperms, monocots, dicots, C3, C4, CAM plants, bryophytes, ferns, and / or fern allies, microalgae, and / or macroalgae. The plants and / or plant parts useful for the present invention can be plants or plant parts of any plant species / variety / cultivar. As used herein, the term "plant part" includes, but is not limited to, embryo, pollen, ovule, seed, leaf, stem, shoot, flower, branch, fruit, grain, spike, rachis, husk, stalk, root, root tip, anther, plant cell (including plant cells that are intact in a plant and / or plant part), plant protoplast, plant tissue, plant cell tissue culture, plant callus, plant clump, etc. As used herein, a "shoot" refers to the above-ground portion including leaves and stem. Further, as used herein, a "plant cell" refers to the structural and physiological unit of a plant that contains a cell wall and can also refer to a protoplast. A plant cell can be in the form of an isolated single cell, or can be a cultured cell, or can be part of a more highly organized unit (e.g., a plant tissue or plant organ).

[0170] The fusion proteins, polynucleotides, and nucleic acid constructs of the present invention can be used to modify target nucleic acids in any plant or plant part (e.g., base editing, cleavage, nick generation, etc.). Non-limiting examples of plants useful for the present invention include: turfgrasses (e.g., Poa pratensis, Agrostis stolonifera, Lolium perenne, Festuca arundinacea), Calamagrostis x acutiflora, Deschampsia cespitosa, Miscanthus, Arundo donax, Panicum virgatum, vegetable crops, including artichoke, kohlrabi, arugula, leek, asparagus, lettuce (e.g., head lettuce, leaf lettuce, romaine lettuce), taro, melons (e.g., cantaloupe, watermelon, Crenshaw melon, honeydew melon, Roma melon), brassica crops (e.g., Brussels sprouts, cabbage, cauliflower, kale, collard greens, turnip greens, Chinese cabbage, pak choi), cardoon, carrot, Chinese cabbage (napa), okra, onion, celery, parsley, chickpea, parsnip, chicory, pepper, potato, cucurbits (e.g., zucchini, cucumber, pattypan squash, butternut squash, pumpkin, honeydew melon, watermelon, Roma melon), radish, dry bulb onion, rutabaga, eggplant, salsify, endive, scallions, mache, garlic, spinach, green onion, summer squash, leafy greens, beets (sugar beet and fodder beet), sweet potato, Swiss chard, horseradish, tomato, turnip, and spices; fruit crops, such as apple, apricot, cherry, nectarine, peach, pear, plum, prune, cherry, quince, fig, nuts (e.g., chestnut, pecan, pistachio, hazelnut, pistachio, peanut, walnut, macadamia nut, almond, etc.), citrus (e.g., clementine, kumquat, orange, grapefruit, tangerine, mandarin, lemon, lime, etc.), blueberry, black raspberry, boysenberry, cranberry, currant, gooseberry, loganberry, raspberry, strawberry, blackberry, grape (wine grape and table grape), avocado, banana, kiwi, persimmon, pomegranate, pineapple, tropical fruits, pome fruits, melons, mango, papaya, and lychee, field crop plants such as clover, alfalfa, timothy, evening primrose, meadow foam, corn / maize (field corn, sweet corn, popcorn), hops, jojoba, buckwheat, safflower, quinoa, wheat, rice, barley, rye, millet, sorghum, oats, triticale, sorghum, tobacco, kapok, legumes (beans (e.g., green bean and dry bean), lentil, pea, soybean), oil plants (rapeseed, canola, mustard, poppy, olive, sunflower, coconut, castor oil plant, cocoa bean, peanut, oil palm), duckweed, Arabidopsis, fiber plants (cotton, flax, hemp, jute), Cannabis (e.g., Cannabis sativa, Cannabis indica, and Cannabis ruderalis), Lauraceae (cinnamon, camphor tree), or plants such as coffee, sugarcane, tea, and natural rubber plants;And / or flower bed plants such as flowering plants, cacti, succulents and / or ornamental plants (e.g., roses, tulips, violets), and trees such as forest trees (broad-leaved trees and evergreen trees, e.g., conifers; e.g., elm, ash, oak, maple, fir, spruce, cedar, pine, birch, cypress, eucalyptus, willow), and shrubs and other nursery stock. In some embodiments, the fusion proteins of the invention and the polynucleotides and nucleic acid constructs encoding them can be used to modify maize, soybean, wheat, canola, rice, tomato, pepper, sunflower, raspberry, blackberry, black raspberry and / or cherry.;

[0171] The invention further includes a kit for implementing the methods of the invention. The kits of the invention can contain reagents, buffers and instruments for mixing, measuring, sorting, labeling, etc., and instructions, etc., as would be suitable for modifying target nucleic acids.

[0172] In some embodiments, the invention provides a kit that contains one or more polypeptides of the invention, one or more fusion proteins of the invention, one or more polynucleotides encoding one or more fusion proteins of the invention, the CRISPR-Cas system of the invention and / or an expression cassette or vector containing the same, and optionally instructions regarding its use. In some embodiments, the kit can further contain: a Cas12a guide nucleic acid and / or an expression cassette or vector containing the same. In some embodiments, the guide nucleic acid can be provided on the same expression cassette or vector as the polynucleotide encoding the fusion protein of the invention.

[0173] Thus, in some embodiments, a kit is provided that contains a nucleic acid construct that includes: (a) a polynucleotide encoding a fusion protein provided herein; and (b) a promoter driving the expression of the polynucleotide of (a). In some embodiments, the kit can further contain a nucleic acid construct encoding a guide nucleic acid, wherein the construct contains a cloning site for cloning a nucleic acid sequence identical or complementary to a target nucleic acid sequence into the backbone of the guide nucleic acid.

[0174] In some embodiments, the polypeptide of the kit can further contain one or more nuclear localization signals fused to the fusion protein, or a polynucleotide encoding the same. In some embodiments, the polynucleotide of the kit can further encode one or more selectable markers useful for identifying transformants (e.g., nucleic acids encoding antibiotic resistance genes, herbicide resistance genes, etc.). In some embodiments, the polynucleotide can be mRNA, which can encode one or more introns within the encoded fusion protein.

[0175] The present invention will now be described with reference to the following examples. It should be appreciated that these examples are not intended to limit the scope of the claims of the present invention, but are intended as examples of certain embodiments. Any variations that occur to those skilled in the art in the methods exemplified are intended to fall within the scope of the present invention. Example

[0176] Example 1

[0177] Although some variants of the Cas12a-based cytosine base editor have been tested, they have lower activity compared to the Cas9-based versions. All the variants tested used the same set of linkers (GS linker, XTEN linker, and GS-XTEN-GS linker) used in the Cas9-based cytosine base editor, and none were rationally or computationally optimized using structure-based techniques. Therefore, we sought to develop an optimized Cas12a-based cytosine base editor by designing the optimal linker lengths and sequences for various domain constructs based on the ideal arrangement of the rAPOBEC1 and UGI domains.

[0178] The initial fusion protein design used the Lachnospiraceae bacterium ND2006 Cas12a (LbCas12a) (e.g., SEQ ID NO: 29) due to its lower temperature sensitivity and proven activity in plant cells; however, due to the high level of structural similarity between different Cas12a endonucleases, these designs should be extended to Cas12a enzymes from other species (e.g., Acidaminococcus sp. Cpf1 (AsCpf1), Francisella novicida Cpf1 (FnCpf1), and others, see, e.g., SEQ ID NOs: 30-45).

[0179] Using a structure-based approach, we developed several linker sequences designed to enable an optimal arrangement of the cytosine deaminase domain relative to Cas12a such that they will be able to access the single-stranded portion of the non-target strand for base editing. We also designed the linker sequences to ensure that the UGI domain is positioned such that it can bind to uracil-DNA glycosylase without interfering with the other components of the base editor. Due to the arrangement of the termini of Cas12a and the orientation of its guide RNA, the ideal linkers for these arrangements differ significantly from the prior art linkers used in Cas9 CBEs as well as the published versions of Cas12a-based CBEs. These linkers were designed to accommodate several possible base editor domain architectures where the deaminase domain is linked to either terminus of Cas12a. Exemplary designed linkers are provided in Table 1.

[0180] Table 1. Exemplary linkers

[0181]

[0182]

[0183] To test the effectiveness of each designed linker sequence (including length, flexibility, and susceptibility to proteases), constructs were generated that contain each linker sequence for a specific architecture in a vector for expression in mammalian cells. The domain arrangements of Cas12a-based cytosine base editors selected for experimental screening are provided in Figure 1 A-C, and exemplary sequences generated for testing include SEQ ID NO:49 - 72.

[0184] In the case of constructs with the domain arrangements illustrated in Figure 1 A and Figure 1 C, the linkers to APOBEC1 and UGI are independent of each other relative to Cas12a; thus, they were placed in separate constructs and paired with control linkers of matching length (8-residue GS, XTEN, or GS-XTEN-GS). For the domain arrangement illustrated in Figure 1 C, since both linkers potentially affect the position of the deaminase, all combinations of the designed linkers were tested. Two previously tested Cas12a cytosine base editor designs were used as controls ( Figure 2 ).

[0185] After screening in mammalian cells, the most effective linker for each architecture was selected for testing in stable plant transformation (e.g., soybean).

[0186] Example 2

[0187] HEK293T cell test

[0188] HEK293T cells (human cell line) were seeded into a collagen-coated 48-well plate (Corning) without antibiotics and using DMEM (Dulbecco's Modified Eagle Medium) medium. At 70 - 80% confluence, cells were transfected with 1.5 μL of Lipofectamine 3000 (ThermoFisher Scientific) using 750 ng of base editor plasmid and 250 ng of guide RNA plasmid according to the manufacturer's experimental protocol. After 3 days, the cells were lysed and DNA was extracted using MagMax TM DNA extraction kit (Applied Biosystems).

[0189] As described above, all constructs listed in Table 1 were tested in HEK293T cells in a total of four experiments. Results for each of the four experiments are provided in Tables 2 - 5 below and in Figure 3 - 20 . The percentages in Tables 2 - 5 indicate the maximum amount of C->T editing observed at any base in the indicated spacer.

[0190] In Figure 3 - 20 , the constructs are listed on the x-axis, where each bar for a given construct represents the editing at a single cytosine within the editing window, as described in the legend. The y-axis indicates the editing level observed for each cytosine within the window. Error bars represent the standard deviation between multiple experiments for the same construct and guide. When no error bars are present, only one set of measurements was taken. For some constructs and spacers (labeled "ND = no data" in the table), the editing efficiency was not determined.

[0191] The designed linkers (especially in the ACU configuration) showed improved editing efficiency relative to the control constructs.

[0192] The spacer sequences are as follows:

[0193] EMX1 spacer 1: TCATCTGTGCCCCTCCCTCCCTG (SEQ ID NO:83)

[0194] RUNX1 spacer 1: AGCCTCACCCCTCTAGCCCTACA (SEQ ID NO:84)

[0195] RUNX1 Spacer 2: TTCTCCCCTCTGCTGGATACCTC (SEQ ID NO:85)

[0196] DNMT1 Spacer 1: CCTCACTCCTGCTCGGTGAATTT (SEQ ID NO:86)

[0197] DNMT1 Spacer 2: GCTCAGCAGGCACCTGCCTCAGC (SEQ ID NO:87)

[0198] AAVS1 Spacer 1: TCTGTCCCCTCCACCCCACAGTG (SEQ ID NO:88)

[0199] Table 2: Initial editing results for Cas12a CBE constructs as Apobec1 fusions

[0200]

[0201] Table 3: Initial editing results for the remaining Cas12a CBE constructs as Apobec1 fusions

[0202]

[0203]

[0204] Table 4: Repeated editing results for Cas12a CBE constructs

[0205]

[0206] Table 5: Repeated editing results for the remaining Cas12a CBE constructs

[0207]

[0208]

[0209] * This construct was tested as a fusion with A3A (instead of Apobec1).

[0210] Example 3

[0211] In addition, a subgroup of designs was tested as fusions with human A3A, a highly active deaminase that has previously been shown to enable efficient cytosine base editing. To stabilize these constructs, introns were included in the sequence of A3A. Two additional constructs, UCA_L2_2R and UCA_L2_4, were too unstable to be purified as Apobec1 fusions without introns and are therefore shown here as fusions with Apobec1 that contain introns in the coding region. For Figure 21 - 25 , the Y-axis indicates the C-to-T editing efficiency obtained at each of the cytosines shown in the legend, and the constructs are listed on the X-axis, with each cytosine within the spacer depicted by a different bar. When "ND" is indicated, it means that no data were collected for the specified sample and spacer.

[0212] Results for this example are provided in Table 6 below and in Figure 21 - 25 . The percentages in Table 6 indicate the maximum amount of C->T editing observed at any base in the indicated spacer.

[0213] Table 6

[0214]

[0215] *A3A = APOBEC3A

[0216] Exemplary editor construction

[0217] ACU_L1_5R A3A (HCF version): SEQ ID NO:91, 93

[0218] ACU_L1_5R A3A (soybean version): SEQ ID NO:92, 94

[0219] ACU_L1_2 A3A (HCF version): SEQ ID NO:95, 96

[0220] ACU_L1_3R A3A (HCF version): SEQ ID NO:97, 98

[0221] UCA_L2_1 A3A (HCF version): SEQ ID NO:99, 100

[0222] CUA control: SEQ ID NO:101

[0223] ACU control: SEQ ID NO:102

[0224] Shanghai Tech Control: SEQ ID NO:103

[0225] ACU Control A3A (HCF version): SEQ ID NO:104, 105

[0226] UCA_L2_2R (Apobec1 + intron): SEQ ID NO:106

[0227] UCA_L2_4 (Apobec1 + intron): SEQ ID NO:107

[0228] Example 4

[0229] R - SODA Experimental Protocol

[0230] For the Rapid and Stable Soybean Assay (R - SODA), by using sonication, Agrobacterium tumefaciens containing a plasmid with a suitable construct and guide cassette encoded in its T - DNA was used to infiltrate rehydrated and dried soybean explants. The explants were co - cultured with Agrobacterium tumefaciens for four days and transferred to a selection medium. Then, they were cultured on the selection medium for four weeks and the seedlings were collected for screening. Editing in each sampled seedling was evaluated by using next - generation sequencing.

[0231] Using the editor constructs of the present invention (ACU_L1_5R(A3A), ACU_L1_5R, ACU_L1_2), three different nucleic acid targets (locus 1, locus 2, locus 3) in soybeans were edited. The results are shown in Figure 26 in.

[0232] The above content illustrates the present invention and should not be construed as limiting the present invention. The present invention is defined by the following claims, which include equivalents of these claims. Sequence Listing <110> Pairwise Plants Services, Inc Guffy, Sharon Leigh Watts, Joseph Matthew <120> Optimized Protein Linkers and Methods of Use <130> 1499 - 6WO <150> US 62 / 876,275 <151> 2019 - 07 - 19 <160> 107 <170> PatentIn version 3.5 <210> 1 <211> 36 <212> PRT <213> Artificial <220> <223> Connector <400> 1 Glu Lys Ser Lys Asn Asp Arg Ser Lys Pro Gln Pro Ser Asp Asp Arg 1 5 10 15 Asp Arg Gln Pro Pro Ser Gly Glu Asp Tyr Pro Glu Trp Lys Ala Pro 20 25 30 Gly Glu Tyr Glu 35 <210> 2 <211> 34 <212> PRT <213> Artificial <220> <223> Connector <400> 2 Gln Glu Pro Lys Pro Gln Asp Gln Ser Ser Glu Val Pro Pro Pro Pro 1 5 10 15 Gly Ser Gln Lys Pro Gly Thr Lys Glu Pro His Asp Ser Lys Ser Ser 20 25 30 Gly Pro <210> 3 <211> 34 <212> PRT <213> Artificial <220> <223> Connector <400> 3 Pro Asp Asn Ser Ser Gly Gln Lys Leu Gln Leu Pro Gln Pro Ser Asp 1 5 10 15 Lys Pro Gln Asp Ser Arg Glu Lys Ser Asp Ser Leu Pro Ser Asp Lys 20 25 30 Arg Asp <210> 4 <211> 36 <212> PRT <213> Artificial <220> <223> Linker <400> 4 Pro Asp Asn Ser Thr Leu Gln Thr Leu Gln Leu Pro Gln Pro Thr Pro 1 5 10 15 Ser Ser Thr Asp Thr Gln Gln Thr Ser Asp Thr Asp Pro Glu Asp Thr 20 25 30 Thr Asp Val Ile 35 <210> 5 <211> 30 <212> PRT <213> Artificial <220> <223> Linker <400> 5 Ser Thr Ser Gln Ser Asp Gly Ser Ser Val Pro Ala Asp Ile Asp Gln 1 5 10 15 Ser Ser Asp Ser Asp Gln Ser Ser Ser Gln Gly Gln Pro Gly 20 25 30 <210> 6 <211> 14 <212> PRT <213> Artificial <220> <223> Linker <400> 6 Ala Lys Pro Asp Asp Glu Ser Gln Lys Pro Pro Gln Asp Asp 1 5 10 <210> 7 <211> 14 <212> PRT <213> Artificial <220> <223> Linker <400> 7 Leu Gln Leu Glu Pro Gly Pro Thr Thr Pro Glu Tyr Pro Ile 1 5 10 <210> 8 <211> 14 <212> PRT <213> Artificial <220> <223> Linker <400> 8 Ile Gln Leu Pro Pro Ser Asp Thr Thr Pro Glu Asn Pro Ile 1 5 10 <210> 9 <211> 12 <212> PRT <213> Artificial <220> <223> Linker <400> 9 Glu Ser Asn Asp Asn Ser Gln Val Pro Pro Ser Leu 1 5 10 <210> 10 <211> 10 <212> PRT <213> Artificial <220> <223> Linker <400> 10 Ser Glu Gln Gln Glu Tyr Pro Gly Ser Gly 1 5 10 <210> 11 <211> 10 <212> PRT <213> Artificial <220> <223> Connector <400> 11 Asn Asn Ser Glu Gln Gln Glu Asn Pro Ala 1 5 10 <210> 12 <211> 12 <212> PRT <213> Artificial <220> <223> Connector <400> 12 Ser Thr Asp Gly Ser Gly Gln Pro Lys His Lys Pro 1 5 10 <210> 13 <211> 20 <212> PRT <213> Artificial <220> <223> Connector <400> 13 Pro Lys Pro Ser Ser Glu Ser Gly Glu Arg Tyr Glu Gln Gln Pro Glu 1 5 10 15 Pro Pro Pro Pro 20 <210> 14 <211> 16 <212> PRT <213> Artificial <220> <223> Connector <400> 14 Lys Gly Gly Gly Gly Glu Pro Asp Glu Lys Arg Pro Ser Gln Ser Ser 1 5 10 15 <210> 15 <211> 14 <212> PRT <213> Artificial <220> <223> Connector <400> 15 Tyr Ala Gly Gly Thr Pro Lys Glu Pro Pro Pro Pro Asn Ser 1 5 10 <210> 16 <211> 14 <212> PRT <213> Artificial <220> <223> Connector <400> 16 Pro Leu Val Ala Gly Gly Thr Pro Phe Glu Pro Pro Pro Pro 1 5 10 <210> 17 <211> 14 <212> PRT <213> Artificial <220> <223> Connector <400> 17 Pro Gln Pro Asp Glu Arg Ser Gln Ile Pro Asp Asn Lys Glu 1 5 10 <210> 18 <211> 10 <212> PRT <213> Artificial <220> <223> Connector <400> 18 Tyr Thr Asp Glu Lys Pro Leu Pro Arg Ser 1 5 10 <210> 19 <211> 12 <212> PRT <213> Artificial <220> <223> Connector <400> 19 Ser His Pro Pro Gln Glu Pro Pro Gln Ser Asn Leu 1 5 10 <210> 20 <211> 16 <212> PRT <213> Artificial <220> <223> Connector <400> 20 Ser Glu Ser Pro Ser Lys Gln Gln Pro Glu Pro Lys Ser Ser Lys Gly 1 5 10 15 <210> 21 <211> 16 <212> PRT <213> Artificial <220> <223> Connector <400> 21 Ser Glu Ser Pro Thr Asn Gln Gln Pro Glu Pro Gln Trp Thr Thr Asp 1 5 10 15 <210> 22 <211> 16 <212> PRT <213> Artificial <220> <223> Connector <400> 22 Gly Gly Ser Lys Gly Pro Pro Pro Ser Pro Pro Pro Pro Gln Pro Glu 1 5 10 15 <210> 23 <211> 16 <212> PRT <213> Artificial <220> <223> Connector <400> 23 Gly Pro Leu Pro Ala Pro Pro Pro Gln Pro Pro Pro Pro Gln Pro Asn 1 5 10 15 <210> 24 <211> 14 <212> PRT <213> Artificial <220> <223> Connector <400> 24 Arg Pro Leu Pro His Asp Asn Asn Lys Gln Asp Tyr Ser Lys 1 5 10 <210> 25 <211> 4 <212> PRT <213> Artificial <220> <223> Connector <400> 25 Ser Gly Gly Ser 1 <210> 26 <211> 10 <212> PRT <213> Artificial <220> <223> Connector <400> 26 Ser Gly Gly Ser Gly Gly Ser Gly Gly Ser 1 5 10 <210> 27 <211> 16 <212> PRT <213> Artificial <220> <223> Connector <400> 27 Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser 1 5 10 15 <210> 28 <211> 32 <212> PRT <213> Artificial <220> <223> Connector <400> 28 Ser Gly Gly Ser Ser Gly Gly Ser Ser Gly Ser Glu Thr Pro Gly Thr 1 5 10 15 Ser Glu Ser Ala Thr Pro Glu Ser Ser Gly Gly Ser Ser Gly Gly Ser 20 25 30 <210> 29 <211> 1227 <212> PRT <213> Bacteria of the family Lachnospiraceae <400> 29 Ser Lys Leu Glu Lys Phe Thr Asn Cys Tyr Ser Leu Ser Lys Thr Leu 1 5 10 15 Arg Phe Lys Ala Ile Pro Val Gly Lys Thr Gln Glu Asn Ile Asp Asn 20 25 30 Lys Arg Leu Leu Val Glu Asp Glu Lys Arg Ala Glu Asp Tyr Lys Gly 35 40 45 Val Lys Lys Leu Leu Asp Arg Tyr Tyr Leu Ser Phe Ile Asn Asp Val 50 55 60 Leu His Ser Ile Lys Leu Lys Asn Leu Asn Asn Tyr Ile Ser Leu Phe 65 70 75 80 Arg Lys Lys Thr Arg Thr Glu Lys Glu Asn Lys Glu Leu Glu Asn Leu 85 90 95 Glu Ile Asn Leu Arg Lys Glu Ile Ala Lys Ala Phe Lys Gly Asn Glu 100 105 110 Gly Tyr Lys Ser Leu Phe Lys Lys Asp Ile Ile Glu Thr Ile Leu Pro 115 120 125 Glu Phe Leu Asp Asp Lys Asp Glu Ile Ala Leu Val Asn Ser Phe Asn 130 135 140 Gly Phe Thr Thr Ala Phe Thr Gly Phe Phe Asp Asn Arg Glu Asn Met 145 150 155 160 Phe Ser Glu Glu Ala Lys Ser Thr Ser Ile Ala Phe Arg Cys Ile Asn 165 170 175 Glu Asn Leu Thr Arg Tyr Ile Ser Asn Met Asp Ile Phe Glu Lys Val 180 185 190 Asp Ala Ile Phe Asp Lys His Glu Val Gln Glu Ile Lys Glu Lys Ile 195 200 205 Leu Asn Ser Asp Tyr Asp Val Glu Asp Phe Phe Glu Gly Glu Phe Phe 210 215 220 Asn Phe Val Leu Thr Gln Glu Gly Ile Asp Val Tyr Asn Ala Ile Ile 225 230 235 240 Gly Gly Phe Val Thr Glu Ser Gly Glu Lys Ile Lys Gly Leu Asn Glu 245 250 255 Tyr Ile Asn Leu Tyr Asn Gln Lys Thr Lys Gln Lys Leu Pro Lys Phe 260 265 270 Lys Pro Leu Tyr Lys Gln Val Leu Ser Asp Arg Glu Ser Leu Ser Phe 275 280 285 Tyr Gly Glu Gly Tyr Thr Ser Asp Glu Glu Val Leu Glu Val Phe Arg 290 295 300 Asn Thr Leu Asn Lys Asn Ser Glu Ile Phe Ser Ser Ile Lys Lys Leu 305 310 315 320 Glu Lys Leu Phe Lys Asn Phe Asp Glu Tyr Ser Ser Ala Gly Ile Phe 325 330 335 Val Lys Asn Gly Pro Ala Ile Ser Thr Ile Ser Lys Asp Ile Phe Gly 340 345 350 Glu Trp Asn Val Ile Arg Asp Lys Trp Asn Ala Glu Tyr Asp Asp Ile 355 360 365 His Leu Lys Lys Lys Ala Val Val Thr Glu Lys Tyr Glu Asp Asp Arg 370 375 380 Arg Lys Ser Phe Lys Lys Ile Gly Ser Phe Ser Leu Glu Gln Leu Gln 385 390 395 400 Glu Tyr Ala Asp Ala Asp Leu Ser Val Val Glu Lys Leu Lys Glu Ile 405 410 415 Ile Ile Gln Lys Val Asp Glu Ile Tyr Lys Val Tyr Gly Ser Ser Glu 420 425 430 Lys Leu Phe Asp Ala Asp Phe Val Leu Glu Lys Ser Leu Lys Lys Asn 435 440 445 Asp Ala Val Val Ala Ile Met Lys Asp Leu Leu Asp Ser Val Lys Ser 450 455 460 Phe Glu Asn Tyr Ile Lys Ala Phe Phe Gly Glu Gly Lys Glu Thr Asn 465 470 475 480 Arg Asp Glu Ser Phe Tyr Gly Asp Phe Val Leu Ala Tyr Asp Ile Leu 485 490 495 Leu Lys Val Asp His Ile Tyr Asp Ala Ile Arg Asn Tyr Val Thr Gln 500 505 510 Lys Pro Tyr Ser Lys Asp Lys Phe Lys Leu Tyr Phe Gln Asn Pro Gln 515 520 525 Phe Met Gly Gly Trp Asp Lys Asp Lys Glu Thr Asp Tyr Arg Ala Thr 530 535 540 Ile Leu Arg Tyr Gly Ser Lys Tyr Tyr Leu Ala Ile Met Asp Lys Lys 545 550 555 560 Tyr Ala Lys Cys Leu Gln Lys Ile Asp Lys Asp Asp Val Asn Gly Asn 565 570 575 Tyr Glu Lys Ile Asn Tyr Lys Leu Leu Pro Gly Pro Asn Lys Met Leu 580 585 590 Pro Lys Val Phe Phe Ser Lys Lys Trp Met Ala Tyr Tyr Asn Pro Ser 595 600 605 Glu Asp Ile Gln Lys Ile Tyr Lys Asn Gly Thr Phe Lys Lys Gly Asp 610 615 620 Met Phe Asn Leu Asn Asp Cys His Lys Leu Ile Asp Phe Phe Lys Asp 625 630 635 640 Ser Ile Ser Arg Tyr Pro Lys Trp Ser Asn Ala Tyr Asp Phe Asn Phe 645 650 655 Ser Glu Thr Glu Lys Tyr Lys Asp Ile Ala Gly Phe Tyr Arg Glu Val 660 665 670 Glu Glu Gln Gly Tyr Lys Val Ser Phe Glu Ser Ala Ser Lys Lys Glu 675 680 685 Val Asp Lys Leu Val Glu Glu Gly Lys Leu Tyr Met Phe Gln Ile Tyr 690 695 700 Asn Lys Asp Phe Ser Asp Lys Ser His Gly Thr Pro Asn Leu His Thr 705 710 715 720 Met Tyr Phe Lys Leu Leu Phe Asp Glu Asn Asn His Gly Gln Ile Arg 725 730 735 Leu Ser Gly Gly Ala Glu Leu Phe Met Arg Arg Ala Ser Leu Lys Lys 740 745 750 Glu Glu Leu Val Val His Pro Ala Asn Ser Pro Ile Ala Asn Lys Asn 755 760 765 Pro Asp Asn Pro Lys Lys Thr Thr Thr Leu Ser Tyr Asp Val Tyr Lys 770 775 780 Asp Lys Arg Phe Ser Glu Asp Gln Tyr Glu Leu His Ile Pro Ile Ala 785 790 795 800 Ile Asn Lys Cys Pro Lys Asn Ile Phe Lys Ile Asn Thr Glu Val Arg 805 810 815 Val Leu Leu Lys His Asp Asp Asn Pro Tyr Val Ile Gly Ile Ala Arg 820 825 830 Gly Glu Arg Asn Leu Leu Tyr Ile Val Val Val Asp Gly Lys Gly Asn 835 840 845 Ile Val Glu Gln Tyr Ser Leu Asn Glu Ile Ile Asn Asn Phe Asn Gly 850 855 860 Ile Arg Ile Lys Thr Asp Tyr His Ser Leu Leu Asp Lys Lys Glu Lys 865 870 875 880 Glu Arg Phe Glu Ala Arg Gln Asn Trp Thr Ser Ile Glu Asn Ile Lys 885 890 895 Glu Leu Lys Ala Gly Tyr Ile Ser Gln Val Val His Lys Ile Cys Glu 900 905 910 Leu Val Glu Lys Tyr Asp Ala Val Ile Ala Leu Glu Asp Leu Asn Ser 915 920 925 Gly Phe Lys Asn Ser Arg Val Lys Val Glu Lys Gln Val Tyr Gln Lys 930 935 940 Phe Glu Lys Met Leu Ile Asp Lys Leu Asn Tyr Met Val Asp Lys Lys 945 950 955 960 Ser Asn Pro Cys Ala Thr Gly Gly Ala Leu Lys Gly Tyr Gln Ile Thr 965 970 975 Asn Lys Phe Glu Ser Phe Lys Ser Met Ser Thr Gln Asn Gly Phe Ile 980 985 990 Phe Tyr Ile Pro Ala Trp Leu Thr Ser Lys Ile Asp Pro Ser Thr Gly 995 1000 1005 Phe Val Asn Leu Leu Lys Thr Lys Tyr Thr Ser Ile Ala Asp Ser 1010 1015 1020 Lys Lys Phe Ile Ser Ser Phe Asp Arg Ile Met Tyr Val Pro Glu 1025 1030 1035 Glu Asp Leu Phe Glu Phe Ala Leu Asp Tyr Lys Asn Phe Ser Arg 1040 1045 1050 Thr Asp Ala Asp Tyr Ile Lys Lys Trp Lys Leu Tyr Ser Tyr Gly 1055 1060 1065 Asn Arg Ile Arg Ile Phe Arg Asn Pro Lys Lys Asn Asn Val Phe 1070 1075 1080 Asp Trp Glu Glu Val Cys Leu Thr Ser Ala Tyr Lys Glu Leu Phe 1085 1090 1095 Asn Lys Tyr Gly Ile Asn Tyr Gln Gln Gly Asp Ile Arg Ala Leu 1100 1105 1110 Leu Cys Glu Gln Ser Asp Lys Ala Phe Tyr Ser Ser Phe Met Ala 1115 1120 1125 Leu Met Ser Leu Met Leu Gln Met Arg Asn Ser Ile Thr Gly Arg 1130 1135 1140 Thr Asp Val Asp Phe Leu Ile Ser Pro Val Lys Asn Ser Asp Gly 1145 1150 1155 Ile Phe Tyr Asp Ser Arg Asn Tyr Glu Ala Gln Glu Asn Ala Ile 1160 1165 1170 Leu Pro Lys Asn Ala Asp Ala Asn Gly Ala Tyr Asn Ile Ala Arg 1175 1180 1185 Lys Val Leu Trp Ala Ile Gly Gln Phe Lys Lys Ala Glu Asp Glu 1190 1195 1200 Lys Leu Asp Lys Val Lys Ile Ala Ile Ser Asn Lys Glu Trp Leu 1205 1210 1215 Glu Tyr Ala Gln Thr Ser Val Lys His 1220 1225 <210> 30 <211> 1307 <212> PRT <213> Acidaminococcus sp. <400> 30 Met Thr Gln Phe Glu Gly Phe Thr Asn Leu Tyr Gln Val Ser Lys Thr 1 5 10 15 Leu Arg Phe Glu Leu Ile Pro Gln Gly Lys Thr Leu Lys His Ile Gln 20 25 30 Glu Gln Gly Phe Ile Glu Glu Asp Lys Ala Arg Asn Asp His Tyr Lys 35 40 45 Glu Leu Lys Pro Ile Ile Asp Arg Ile Tyr Lys Thr Tyr Ala Asp Gln 50 55 60 Cys Leu Gln Leu Val Gln Leu Asp Trp Glu Asn Leu Ser Ala Ala Ile 65 70 75 80 Asp Ser Tyr Arg Lys Glu Lys Thr Glu Glu Thr Arg Asn Ala Leu Ile 85 90 95 Glu Glu Gln Ala Thr Tyr Arg Asn Ala Ile His Asp Tyr Phe Ile Gly 100 105 110 Arg Thr Asp Asn Leu Thr Asp Ala Ile Asn Lys Arg His Ala Glu Ile 115 120 125 Tyr Lys Gly Leu Phe Lys Ala Glu Leu Phe Asn Gly Lys Val Leu Lys 130 135 140 Gln Leu Gly Thr Val Thr Thr Thr Glu His Glu Asn Ala Leu Leu Arg 145 150 155 160 Ser Phe Asp Lys Phe Thr Thr Tyr Phe Ser Gly Phe Tyr Glu Asn Arg 165 170 175 Lys Asn Val Phe Ser Ala Glu Asp Ile Ser Thr Ala Ile Pro His Arg 180 185 190 Ile Val Gln Asp Asn Phe Pro Lys Phe Lys Glu Asn Cys His Ile Phe 195 200 205 Thr Arg Leu Ile Thr Ala Val Pro Ser Leu Arg Glu His Phe Glu Asn 210 215 220 Val Lys Lys Ala Ile Gly Ile Phe Val Ser Thr Ser Ile Glu Glu Val 225 230 235 240 Phe Ser Phe Pro Phe Tyr Asn Gln Leu Leu Thr Gln Thr Gln Ile Asp 245 250 255 Leu Tyr Asn Gln Leu Leu Gly Gly Ile Ser Arg Glu Ala Gly Thr Glu 260 265 270 Lys Ile Lys Gly Leu Asn Glu Val Leu Asn Leu Ala Ile Gln Lys Asn 275 280 285 Asp Glu Thr Ala His Ile Ile Ala Ser Leu Pro His Arg Phe Ile Pro 290 295 300 Leu Phe Lys Gln Ile Leu Ser Asp Arg Asn Thr Leu Ser Phe Ile Leu 305 310 315 320 Glu Glu Phe Lys Ser Asp Glu Glu Val Ile Gln Ser Phe Cys Lys Tyr 325 330 335 Lys Thr Leu Leu Arg Asn Glu Asn Val Leu Glu Thr Ala Glu Ala Leu 340 345 350 Phe Asn Glu Leu Asn Ser Ile Asp Leu Thr His Ile Phe Ile Ser His 355 360 365 Lys Lys Leu Glu Thr Ile Ser Ser Ala Leu Cys Asp His Trp Asp Thr 370 375 380 Leu Arg Asn Ala Leu Tyr Glu Arg Arg Ile Ser Glu Leu Thr Gly Lys 385 390 395 400 Ile Thr Lys Ser Ala Lys Glu Lys Val Gln Arg Ser Leu Lys His Glu 405 410 415 Asp Ile Asn Leu Gln Glu Ile Ile Ser Ala Ala Gly Lys Glu Leu Ser 420 425 430 Glu Ala Phe Lys Gln Lys Thr Ser Glu Ile Leu Ser His Ala His Ala 435 440 445 Ala Leu Asp Gln Pro Leu Pro Thr Thr Leu Lys Lys Gln Glu Glu Lys 450 455 460 Glu Ile Leu Lys Ser Gln Leu Asp Ser Leu Leu Gly Leu Tyr His Leu 465 470 475 480 Leu Asp Trp Phe Ala Val Asp Glu Ser Asn Glu Val Asp Pro Glu Phe 485 490 495 Ser Ala Arg Leu Thr Gly Ile Lys Leu Glu Met Glu Pro Ser Leu Ser 500 505 510 Phe Tyr Asn Lys Ala Arg Asn Tyr Ala Thr Lys Lys Pro Tyr Ser Val 515 520 525 Glu Lys Phe Lys Leu Asn Phe Gln Met Pro Thr Leu Ala Ser Gly Trp 530 535 540 Asp Val Asn Lys Glu Lys Asn Asn Gly Ala Ile Leu Phe Val Lys Asn 545 550 555 560 Gly Leu Tyr Tyr Leu Gly Ile Met Pro Lys Gln Lys Gly Arg Tyr Lys 565 570 575 Ala Leu Ser Phe Glu Pro Thr Glu Lys Thr Ser Glu Gly Phe Asp Lys 580 585 590 Met Tyr Tyr Asp Tyr Phe Pro Asp Ala Ala Lys Met Ile Pro Lys Cys 595 600 605 Ser Thr Gln Leu Lys Ala Val Thr Ala His Phe Gln Thr His Thr Thr 610 615 620 Pro Ile Leu Leu Ser Asn Asn Phe Ile Glu Pro Leu Glu Ile Thr Lys 625 630 635 640 Glu Ile Tyr Asp Leu Asn Asn Pro Glu Lys Glu Pro Lys Lys Phe Gln 645 650 655 Thr Ala Tyr Ala Lys Lys Thr Gly Asp Gln Lys Gly Tyr Arg Glu Ala 660 665 670 Leu Cys Lys Trp Ile Asp Phe Thr Arg Asp Phe Leu Ser Lys Tyr Thr 675 680 685 Lys Thr Thr Ser Ile Asp Leu Ser Ser Leu Arg Pro Ser Ser Gln Tyr 690 695 700 Lys Asp Leu Gly Glu Tyr Tyr Ala Glu Leu Asn Pro Leu Leu Tyr His 705 710 715 720 Ile Ser Phe Gln Arg Ile Ala Glu Lys Glu Ile Met Asp Ala Val Glu 725 730 735 Thr Gly Lys Leu Tyr Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ala Lys 740 745 750 Gly His His Gly Lys Pro Asn Leu His Thr Leu Tyr Trp Thr Gly Leu 755 760 765 Phe Ser Pro Glu Asn Leu Ala Lys Thr Ser Ile Lys Leu Asn Gly Gln 770 775 780 Ala Glu Leu Phe Tyr Arg Pro Lys Ser Arg Met Lys Arg Met Ala His 785 790 795 800 Arg Leu Gly Glu Lys Met Leu Asn Lys Lys Leu Lys Asp Gln Lys Thr 805 810 815 Pro Ile Pro Asp Thr Leu Tyr Gln Glu Leu Tyr Asp Tyr Val Asn His 820 825 830 Arg Leu Ser His Asp Leu Ser Asp Glu Ala Arg Ala Leu Leu Pro Asn 835 840 845 Val Ile Thr Lys Glu Val Ser His Glu Ile Ile Lys Asp Arg Arg Phe 850 855 860 Thr Ser Asp Lys Phe Phe Phe His Val Pro Ile Thr Leu Asn Tyr Gln 865 870 875 880 Ala Ala Asn Ser Pro Ser Lys Phe Asn Gln Arg Val Asn Ala Tyr Leu 885 890 895 Lys Glu His Pro Glu Thr Pro Ile Ile Gly Ile Asp Arg Gly Glu Arg 900 905 910 Asn Leu Ile Tyr Ile Thr Val Ile Asp Ser Thr Gly Lys Ile Leu Glu 915 920 925 Gln Arg Ser Leu Asn Thr Ile Gln Gln Phe Asp Tyr Gln Lys Lys Leu 930 935 940 Asp Asn Arg Glu Lys Glu Arg Val Ala Ala Arg Gln Ala Trp Ser Val 945 950 955 960 Val Gly Thr Ile Lys Asp Leu Lys Gln Gly Tyr Leu Ser Gln Val Ile 965 970 975 His Glu Ile Val Asp Leu Met Ile His Tyr Gln Ala Val Val Val Leu 980 985 990 Glu Asn Leu Asn Phe Gly Phe Lys Ser Lys Arg Thr Gly Ile Ala Glu 995 1000 1005 Lys Ala Val Tyr Gln Gln Phe Glu Lys Met Leu Ile Asp Lys Leu 1010 1015 1020 Asn Cys Leu Val Leu Lys Asp Tyr Pro Ala Glu Lys Val Gly Gly 1025 1030 1035 Val Leu Asn Pro Tyr Gln Leu Thr Asp Gln Phe Thr Ser Phe Ala 1040 1045 1050 Lys Met Gly Thr Gln Ser Gly Phe Leu Phe Tyr Val Pro Ala Pro 1055 1060 1065 Tyr Thr Ser Lys Ile Asp Pro Leu Thr Gly Phe Val Asp Pro Phe 1070 1075 1080 Val Trp Lys Thr Ile Lys Asn His Glu Ser Arg Lys His Phe Leu 1085 1090 1095 Glu Gly Phe Asp Phe Leu His Tyr Asp Val Lys Thr Gly Asp Phe 1100 1105 1110 Ile Leu His Phe Lys Met Asn Arg Asn Leu Ser Phe Gln Arg Gly 1115 1120 1125 Leu Pro Gly Phe Met Pro Ala Trp Asp Ile Val Phe Glu Lys Asn 1130 1135 1140 Glu Thr Gln Phe Asp Ala Lys Gly Thr Pro Phe Ile Ala Gly Lys 1145 1150 1155 Arg Ile Val Pro Val Ile Glu Asn His Arg Phe Thr Gly Arg Tyr 1160 1165 1170 Arg Asp Leu Tyr Pro Ala Asn Glu Leu Ile Ala Leu Leu Glu Glu 1175 1180 1185 Lys Gly Ile Val Phe Arg Asp Gly Ser Asn Ile Leu Pro Lys Leu 1190 1195 1200 Leu Glu Asn Asp Asp Ser His Ala Ile Asp Thr Met Val Ala Leu 1205 1210 1215 Ile Arg Ser Val Leu Gln Met Arg Asn Ser Asn Ala Ala Thr Gly 1220 1225 1230 Glu Asp Tyr Ile Asn Ser Pro Val Arg Asp Leu Asn Gly Val Cys 1235 1240 1245 Phe Asp Ser Arg Phe Gln Asn Pro Glu Trp Pro Met Asp Ala Asp 1250 1255 1260 Ala Asn Gly Ala Tyr His Ile Ala Leu Lys Gly Gln Leu Leu Leu 1265 1270 1275 Asn His Leu Lys Glu Ser Lys Asp Leu Lys Leu Gln Asn Gly Ile 1280 1285 1290 Ser Asn Gln Asp Trp Leu Ala Tyr Ile Gln Glu Leu Arg Asn 1295 1300 1305 <210> 31 <211> 1241 <212> PRT <213> Butyrivibrio proteoclasticus <400> 31 Met Leu Leu Tyr Glu Asn Tyr Thr Lys Arg Asn Gln Ile Thr Lys Ser 1 5 10 15 Leu Arg Leu Glu Leu Arg Pro Gln Gly Lys Thr Leu Arg Asn Ile Lys 20 25 30 Glu Leu Asn Leu Leu Glu Gln Asp Lys Ala Ile Tyr Ala Leu Leu Glu 35 40 45 Arg Leu Lys Pro Val Ile Asp Glu Gly Ile Lys Asp Ile Ala Arg Asp 50 55 60 Thr Leu Lys Asn Cys Glu Leu Ser Phe Glu Lys Leu Tyr Glu His Phe 65 70 75 80 Leu Ser Gly Asp Lys Lys Ala Tyr Ala Lys Glu Ser Glu Arg Leu Lys 85 90 95 Lys Glu Ile Val Lys Thr Leu Ile Lys Asn Leu Pro Glu Gly Ile Gly 100 105 110 Lys Ile Ser Glu Ile Asn Ser Ala Lys Tyr Leu Asn Gly Val Leu Tyr 115 120 125 Asp Phe Ile Asp Lys Thr His Lys Asp Ser Glu Glu Lys Gln Asn Ile 130 135 140 Leu Ser Asp Ile Leu Glu Thr Lys Gly Tyr Leu Ala Leu Phe Ser Lys 145 150 155 160 Phe Leu Thr Ser Arg Ile Thr Thr Leu Glu Gln Ser Met Pro Lys Arg 165 170 175 Val Ile Glu Asn Phe Glu Ile Tyr Ala Ala Asn Ile Pro Lys Met Gln 180 185 190 Asp Ala Leu Glu Arg Gly Ala Val Ser Phe Ala Ile Glu Tyr Glu Ser 195 200 205 Ile Cys Ser Val Asp Tyr Tyr Asn Gln Ile Leu Ser Gln Glu Asp Ile 210 215 220 Asp Ser Tyr Asn Arg Leu Ile Ser Gly Ile Met Asp Glu Asp Gly Ala 225 230 235 240 Lys Glu Lys Gly Ile Asn Gln Thr Ile Ser Glu Lys Asn Ile Lys Ile 245 250 255 Lys Ser Glu His Leu Glu Glu Lys Pro Phe Arg Ile Leu Lys Gln Leu 260 265 270 His Lys Gln Ile Leu Glu Glu Arg Glu Lys Ala Phe Thr Ile Asp His 275 280 285 Ile Asp Ser Asp Glu Glu Val Val Gln Val Thr Lys Glu Ala Phe Glu 290 295 300 Gln Thr Lys Glu Gln Trp Glu Asn Ile Lys Lys Ile Asn Gly Phe Tyr 305 310 315 320 Ala Lys Asp Pro Gly Asp Ile Thr Leu Phe Ile Val Val Gly Pro Asn 325 330 335 Gln Thr His Val Leu Ser Gln Leu Ile Tyr Gly Glu His Asp Arg Ile 340 345 350 Arg Leu Leu Leu Glu Glu Tyr Glu Lys Asn Thr Leu Glu Val Leu Pro 355 360 365 Arg Arg Thr Lys Ser Glu Asp Ala Arg Tyr Asp Lys Phe Val Asn Ala 370 375 380 Val Pro Lys Lys Val Ala Lys Glu Ser His Thr Phe Asp Gly Leu Gln 385 390 395 400 Lys Met Thr Gly Asp Asp Arg Leu Phe Ile Leu Tyr Arg Asp Glu Leu 405 410 415 Ala Arg Asn Tyr Met Arg Ile Lys Glu Ala Tyr Gly Thr Phe Glu Arg 420 425 430 Asp Ile Leu Lys Ser Arg Arg Gly Ile Lys Gly Asn Arg Asp Val Gln 435 440 445 Glu Ser Leu Val Ser Phe Tyr Asp Glu Leu Thr Lys Phe Arg Ser Ala 450 455 460 Leu Arg Ile Ile Asn Ser Gly Asn Asp Glu Lys Ala Asp Pro Ile Phe 465 470 475 480 Tyr Asn Thr Phe Asp Gly Ile Phe Glu Lys Ala Asn Arg Thr Tyr Lys 485 490 495 Ala Glu Asn Leu Cys Arg Asn Tyr Val Thr Lys Ser Pro Ala Asp Asp 500 505 510 Ala Arg Ile Met Ala Ser Cys Leu Gly Thr Pro Ala Arg Leu Arg Thr 515 520 525 His Trp Trp Asn Gly Glu Glu Asn Phe Ala Ile Asn Asp Val Ala Met 530 535 540 Ile Arg Arg Gly Asp Glu Tyr Tyr Tyr Phe Val Leu Thr Pro Asp Val 545 550 555 560 Lys Pro Val Asp Leu Lys Thr Lys Asp Glu Thr Asp Ala Gln Ile Phe 565 570 575 Val Gln Arg Lys Gly Ala Lys Ser Phe Leu Gly Leu Pro Lys Ala Leu 580 585 590 Phe Lys Cys Ile Leu Glu Pro Tyr Phe Glu Ser Pro Glu His Lys Asn 595 600 605 Asp Lys Asn Cys Val Ile Glu Glu Tyr Val Ser Lys Pro Leu Thr Ile 610 615 620 Asp Arg Arg Ala Tyr Asp Ile Phe Lys Asn Gly Thr Phe Lys Lys Thr 625 630 635 640 Asn Ile Gly Ile Asp Gly Leu Thr Glu Glu Lys Phe Lys Asp Asp Cys 645 650 655 Arg Tyr Leu Ile Asp Val Tyr Lys Glu Phe Ile Ala Val Tyr Thr Arg 660 665 670 Tyr Ser Cys Phe Asn Met Ser Gly Leu Lys Arg Ala Asp Glu Tyr Asn 675 680 685 Asp Ile Gly Glu Phe Phe Ser Asp Val Asp Thr Arg Leu Cys Thr Met 690 695 700 Glu Trp Ile Pro Val Ser Phe Glu Arg Ile Asn Asp Met Val Asp Lys 705 710 715 720 Lys Glu Gly Leu Leu Phe Leu Val Arg Ser Met Phe Leu Tyr Asn Arg 725 730 735 Pro Arg Lys Pro Tyr Glu Arg Thr Phe Ile Gln Leu Phe Ser Asp Ser 740 745 750 Asn Met Glu His Thr Ser Met Leu Leu Asn Ser Arg Ala Met Ile Gln 755 760 765 Tyr Arg Ala Ala Ser Leu Pro Arg Arg Val Thr His Lys Lys Gly Ser 770 775 780 Ile Leu Val Ala Leu Arg Asp Ser Asn Gly Glu His Ile Pro Met His 785 790 795 800 Ile Arg Glu Ala Ile Tyr Lys Met Lys Asn Asn Phe Asp Ile Ser Ser 805 810 815 Glu Asp Phe Ile Met Ala Lys Ala Tyr Leu Ala Glu His Asp Val Ala 820 825 830 Ile Lys Lys Ala Asn Glu Asp Ile Ile Arg Asn Arg Arg Tyr Thr Glu 835 840 845 Asp Lys Phe Phe Leu Ser Leu Ser Tyr Thr Lys Asn Ala Asp Ile Ser 850 855 860 Ala Arg Thr Leu Asp Tyr Ile Asn Asp Lys Val Glu Glu Asp Thr Gln 865 870 875 880 Asp Ser Arg Met Ala Val Ile Val Thr Arg Asn Leu Lys Asp Leu Thr 885 890 895 Tyr Val Ala Val Val Asp Glu Lys Asn Asn Val Leu Glu Glu Lys Ser 900 905 910 Leu Asn Glu Ile Asp Gly Val Asn Tyr Arg Glu Leu Leu Lys Glu Arg 915 920 925 Thr Lys Ile Lys Tyr His Asp Lys Thr Arg Leu Trp Gln Tyr Asp Val 930 935 940 Ser Ser Lys Gly Leu Lys Glu Ala Tyr Val Glu Leu Ala Val Thr Gln 945 950 955 960 Ile Ser Lys Leu Ala Thr Lys Tyr Asn Ala Val Val Val Val Glu Ser 965 970 975 Met Ser Ser Thr Phe Lys Asp Lys Phe Ser Phe Leu Asp Glu Gln Ile 980 985 990 Phe Lys Ala Phe Glu Ala Arg Leu Cys Ala Arg Met Ser Asp Leu Ser 995 1000 1005 Phe Asn Thr Ile Lys Glu Gly Glu Ala Gly Ser Ile Ser Asn Pro 1010 1015 1020 Ile Gln Val Ser Asn Asn Asn Gly Asn Ser Tyr Gln Asp Gly Val 1025 1030 1035 Ile Tyr Phe Leu Asn Asn Ala Tyr Thr Arg Thr Leu Cys Pro Asp 1040 1045 1050 Thr Gly Phe Val Asp Val Phe Asp Lys Thr Arg Leu Ile Thr Met 1055 1060 1065 Gln Ser Lys Arg Gln Phe Phe Ala Lys Met Lys Asp Ile Arg Ile 1070 1075 1080 Asp Asp Gly Glu Met Leu Phe Thr Phe Asn Leu Glu Glu Tyr Pro 1085 1090 1095 Thr Lys Arg Leu Leu Asp Arg Lys Glu Trp Thr Val Lys Ile Ala 1100 1105 1110 Gly Asp Gly Ser Tyr Phe Asp Lys Asp Lys Gly Glu Tyr Val Tyr 1115 1120 1125 Val Asn Asp Ile Val Arg Glu Gln Ile Ile Pro Ala Leu Leu Glu 1130 1135 1140 Asp Lys Ala Val Phe Asp Gly Asn Met Ala Glu Lys Phe Leu Asp 1145 1150 1155 Lys Thr Ala Ile Ser Gly Lys Ser Val Glu Leu Ile Tyr Lys Trp 1160 1165 1170 Phe Ala Asn Ala Leu Tyr Gly Ile Ile Thr Lys Lys Asp Gly Glu 1175 1180 1185 Lys Ile Tyr Arg Ser Pro Ile Thr Gly Thr Glu Ile Asp Val Ser 1190 1195 1200 Lys Asn Thr Thr Tyr Asn Phe Gly Lys Lys Phe Met Phe Lys Gln 1205 1210 1215 Glu Tyr Arg Gly Asp Gly Asp Phe Leu Asp Ala Phe Leu Asn Tyr 1220 1225 1230 Met Gln Ala Gln Asp Ile Ala Val 1235 1240 <210> 32 <211> 1238 <212> PRT <213> Candidatus Methanoplasma termitum <400> 32 Met Asn Asn Tyr Asp Glu Phe Thr Lys Leu Tyr Pro Ile Gln Lys Thr 1 5 10 15 Ile Arg Phe Glu Leu Lys Pro Gln Gly Arg Thr Met Glu His Leu Glu 20 25 30 Thr Phe Asn Phe Phe Glu Glu Asp Arg Asp Arg Ala Glu Lys Tyr Lys 35 40 45 Ile Leu Lys Glu Ala Ile Asp Glu Tyr His Lys Lys Phe Ile Asp Glu 50 55 60 His Leu Thr Asn Met Ser Leu Asp Trp Asn Ser Leu Lys Gln Ile Ser 65 70 75 80 Glu Lys Tyr Tyr Lys Ser Arg Glu Glu Lys Asp Lys Lys Val Phe Leu 85 90 95 Ser Glu Gln Lys Arg Met Arg Gln Glu Ile Val Ser Glu Phe Lys Lys 100 105 110 Asp Asp Arg Phe Lys Asp Leu Phe Ser Lys Lys Leu Phe Ser Glu Leu 115 120 125 Leu Lys Glu Glu Ile Tyr Lys Lys Gly Asn His Gln Glu Ile Asp Ala 130 135 140 Leu Lys Ser Phe Asp Lys Phe Ser Gly Tyr Phe Ile Gly Leu His Glu 145 150 155 160 Asn Arg Lys Asn Met Tyr Ser Asp Gly Asp Glu Ile Thr Ala Ile Ser 165 170 175 Asn Arg Ile Val Asn Glu Asn Phe Pro Lys Phe Leu Asp Asn Leu Gln 180 185 190 Lys Tyr Gln Glu Ala Arg Lys Lys Tyr Pro Glu Trp Ile Ile Lys Ala 195 200 205 Glu Ser Ala Leu Val Ala His Asn Ile Lys Met Asp Ile Val Phe Ser 210 215 220 Leu Glu Tyr Phe Asn Lys Val Leu Asn Gln Glu Gly Ile Gln Arg Tyr 225 230 235 240 Asn Leu Ala Leu Gly Gly Tyr Val Thr Lys Ser Gly Glu Lys Met Met 245 250 255 Gly Leu Asn Asp Ala Leu Asn Leu Ala His Gln Ser Glu Lys Ser Ser 260 265 270 Lys Gly Arg Ile His Met Thr Pro Leu Phe Lys Gln Ile Leu Ser Glu 275 280 285 Lys Glu Ser Phe Ser Tyr Ile Pro Asp Val Phe Thr Glu Asp Ser Gln 290 295 300 Leu Leu Pro Ser Ile Gly Gly Phe Phe Ala Gln Ile Glu Asn Asp Lys 305 310 315 320 Asp Gly Asn Ile Phe Asp Arg Ala Leu Glu Leu Ile Ser Ser Tyr Ala 325 330 335 Glu Tyr Asp Thr Glu Arg Ile Tyr Ile Arg Gln Ala Asp Ile Asn Arg 340 345 350 Val Ser Asn Val Ile Phe Gly Glu Trp Gly Thr Leu Gly Gly Leu Met 355 360 365 Arg Glu Tyr Lys Ala Asp Ser Ile Asn Asp Ile Asn Leu Glu Arg Thr 370 375 380 Cys Lys Lys Val Asp Lys Trp Leu Asp Ser Lys Glu Phe Ala Leu Ser 385 390 395 400 Asp Val Leu Glu Ala Ile Asp Arg Thr Gly Asn Asn Asp Ala Phe Asn 405 410 415 Glu Tyr Ile Ser Lys Met Arg Thr Ala Arg Glu Lys Ile Asp Ala Ala 420 425 430 Arg Lys Glu Met Lys Phe Ile Ser Glu Lys Ile Ser Gly Asp Glu Glu 435 440 445 Ser Ile His Ile Ile Lys Thr Leu Leu Asp Ser Val Gln Gln Phe Leu 450 455 460 His Phe Phe Asn Leu Phe Lys Ala Arg Gln Asp Ile Pro Leu Asp Gly 465 470 475 480 Ala Phe Tyr Ala Glu Phe Asp Glu Val His Ser Lys Leu Phe Ala Ile 485 490 495 Val Pro Leu Tyr Asn Lys Val Arg Asn Tyr Leu Thr Lys Asn Asn Leu 500 505 510 Asn Thr Lys Lys Ile Lys Leu Asn Phe Lys Asn Pro Thr Leu Ala Asn 515 520 525 Gly Trp Asp Gln Asn Lys Val Tyr Asp Tyr Ala Ser Leu Ile Phe Leu 530 535 540 Arg Asp Gly Asn Tyr Tyr Leu Gly Ile Ile Asn Pro Lys Arg Lys Lys 545 550 555 560 Asn Ile Lys Phe Glu Gln Gly Ser Gly Asn Gly Pro Phe Tyr Arg Lys 565 570 575 Met Val Tyr Lys Gln Ile Pro Gly Pro Asn Lys Asn Leu Arg Pro Val 580 585 590 Phe Leu Thr Ser Thr Lys Gly Lys Lys Glu Tyr Lys Pro Ser Lys Glu 595 600 605 Ile Ile Glu Gly Tyr Glu Ala Asp Lys His Ile Arg Gly Asp Lys Phe 610 615 620 Asp Leu Asp Phe Cys His Lys Leu Ile Asp Phe Phe Lys Glu Ser Ile 625 630 635 640 Glu Lys His Lys Asp Trp Ser Lys Phe Asn Phe Tyr Phe Ser Pro Thr 645 650 655 Glu Ser Tyr Gly Asp Ile Ser Glu Phe Tyr Leu Asp Val Glu Lys Gln 660 665 670 Gly Tyr Arg Met His Phe Glu Asn Ile Ser Ala Glu Thr Ile Asp Glu 675 680 685 Tyr Val Glu Lys Gly Asp Leu Phe Leu Phe Gln Ile Tyr Asn Lys Asp 690 695 700 Phe Val Lys Ala Ala Thr Gly Lys Lys Asp Met His Thr Ile Tyr Trp 705 710 715 720 Asn Ala Ala Phe Ser Pro Glu Asn Leu Gln Asp Val Val Val Lys Leu 725 730 735 Asn Gly Glu Ala Glu Leu Phe Tyr Arg Asp Lys Ser Asp Ile Lys Glu 740 745 750 Ile Val His Arg Glu Gly Glu Ile Leu Val Asn Arg Thr Tyr Asn Gly 755 760 765 Arg Thr Pro Val Pro Asp Lys Ile His Lys Lys Leu Thr Asp Tyr His 770 775 780 Asn Gly Arg Thr Lys Asp Leu Gly Glu Ala Lys Glu Tyr Leu Asp Lys 785 790 795 800 Val Arg Tyr Phe Lys Ala His Tyr Asp Ile Thr Lys Asp Arg Arg Tyr 805 810 815 Leu Asn Asp Lys Ile Tyr Phe His Val Pro Leu Thr Leu Asn Phe Lys 820 825 830 Ala Asn Gly Lys Lys Asn Leu Asn Lys Met Val Ile Glu Lys Phe Leu 835 840 845 Ser Asp Glu Lys Ala His Ile Ile Gly Ile Asp Arg Gly Glu Arg Asn 850 855 860 Leu Leu Tyr Tyr Ser Ile Ile Asp Arg Ser Gly Lys Ile Ile Asp Gln 865 870 875 880 Gln Ser Leu Asn Val Ile Asp Gly Phe Asp Tyr Arg Glu Lys Leu Asn 885 890 895 Gln Arg Glu Ile Glu Met Lys Asp Ala Arg Gln Ser Trp Asn Ala Ile 900 905 910 Gly Lys Ile Lys Asp Leu Lys Glu Gly Tyr Leu Ser Lys Ala Val His 915 920 925 Glu Ile Thr Lys Met Ala Ile Gln Tyr Asn Ala Ile Val Val Met Glu 930 935 940 Glu Leu Asn Tyr Gly Phe Lys Arg Gly Arg Phe Lys Val Glu Lys Gln 945 950 955 960 Ile Tyr Gln Lys Phe Glu Asn Met Leu Ile Asp Lys Met Asn Tyr Leu 965 970 975 Val Phe Lys Asp Ala Pro Asp Glu Ser Pro Gly Gly Val Leu Asn Ala 980 985 990 Tyr Gln Leu Thr Asn Pro Leu Glu Ser Phe Ala Lys Leu Gly Lys Gln 995 1000 1005 Thr Gly Ile Leu Phe Tyr Val Pro Ala Ala Tyr Thr Ser Lys Ile 1010 1015 1020 Asp Pro Thr Thr Gly Phe Val Asn Leu Phe Asn Thr Ser Ser Lys 1025 1030 1035 Thr Asn Ala Gln Glu Arg Lys Glu Phe Leu Gln Lys Phe Glu Ser 1040 1045 1050 Ile Ser Tyr Ser Ala Lys Asp Gly Gly Ile Phe Ala Phe Ala Phe 1055 1060 1065 Asp Tyr Arg Lys Phe Gly Thr Ser Lys Thr Asp His Lys Asn Val 1070 1075 1080 Trp Thr Ala Tyr Thr Asn Gly Glu Arg Met Arg Tyr Ile Lys Glu 1085 1090 1095 Lys Lys Arg Asn Glu Leu Phe Asp Pro Ser Lys Glu Ile Lys Glu 1100 1105 1110 Ala Leu Thr Ser Ser Gly Ile Lys Tyr Asp Gly Gly Gln Asn Ile 1115 1120 1125 Leu Pro Asp Ile Leu Arg Ser Asn Asn Asn Gly Leu Ile Tyr Thr 1130 1135 1140 Met Tyr Ser Ser Phe Ile Ala Ala Ile Gln Met Arg Val Tyr Asp 1145 1150 1155 Gly Lys Glu Asp Tyr Ile Ile Ser Pro Ile Lys Asn Ser Lys Gly 1160 1165 1170 Glu Phe Phe Arg Thr Asp Pro Lys Arg Arg Glu Leu Pro Ile Asp 1175 1180 1185 Ala Asp Ala Asn Gly Ala Tyr Asn Ile Ala Leu Arg Gly Glu Leu 1190 1195 1200 Thr Met Arg Ala Ile Ala Glu Lys Phe Asp Pro Asp Ser Glu Lys 1205 1210 1215 Met Ala Lys Leu Glu Leu Lys His Lys Asp Trp Phe Glu Phe Met 1220 1225 1230 Gln Thr Arg Gly Asp 1235 <210> 33 <211> 1281 <212> PRT <213> Eubacterium eligens <400> 33 Met Asn Gly Asn Arg Ser Ile Val Tyr Arg Glu Phe Val Gly Val Ile 1 5 10 15 Pro Val Ala Lys Thr Leu Arg Asn Glu Leu Arg Pro Val Gly His Thr 20 25 30 Gln Glu His Ile Ile Gln Asn Gly Leu Ile Gln Glu Asp Glu Leu Arg 35 40 45 Gln Glu Lys Ser Thr Glu Leu Lys Asn Ile Met Asp Asp Tyr Tyr Arg 50 55 60 Glu Tyr Ile Asp Lys Ser Leu Ser Gly Val Thr Asp Leu Asp Phe Thr 65 70 75 80 Leu Leu Phe Glu Leu Met Asn Leu Val Gln Ser Ser Pro Ser Lys Asp 85 90 95 Asn Lys Lys Ala Leu Glu Lys Glu Gln Ser Lys Met Arg Glu Gln Ile 100 105 110 Cys Thr His Leu Gln Ser Asp Ser Asn Tyr Lys Asn Ile Phe Asn Ala 115 120 125 Lys Leu Leu Lys Glu Ile Leu Pro Asp Phe Ile Lys Asn Tyr Asn Gln 130 135 140 Tyr Asp Val Lys Asp Lys Ala Gly Lys Leu Glu Thr Leu Ala Leu Phe 145 150 155 160 Asn Gly Phe Ser Thr Tyr Phe Thr Asp Phe Phe Glu Lys Arg Lys Asn 165 170 175 Val Phe Thr Lys Glu Ala Val Ser Thr Ser Ile Ala Tyr Arg Ile Val 180 185 190 His Glu Asn Ser Leu Ile Phe Leu Ala Asn Met Thr Ser Tyr Lys Lys 195 200 205 Ile Ser Glu Lys Ala Leu Asp Glu Ile Glu Val Ile Glu Lys Asn Asn 210 215 220 Gln Asp Lys Met Gly Asp Trp Glu Leu Asn Gln Ile Phe Asn Pro Asp 225 230 235 240 Phe Tyr Asn Met Val Leu Ile Gln Ser Gly Ile Asp Phe Tyr Asn Glu 245 250 255 Ile Cys Gly Val Val Asn Ala His Met Asn Leu Tyr Cys Gln Gln Thr 260 265 270 Lys Asn Asn Tyr Asn Leu Phe Lys Met Arg Lys Leu His Lys Gln Ile 275 280 285 Leu Ala Tyr Thr Ser Thr Ser Phe Glu Val Pro Lys Met Phe Glu Asp 290 295 300 Asp Met Ser Val Tyr Asn Ala Val Asn Ala Phe Ile Asp Glu Thr Glu 305 310 315 320 Lys Gly Asn Ile Ile Gly Lys Leu Lys Asp Ile Val Asn Lys Tyr Asp 325 330 335 Glu Leu Asp Glu Lys Arg Ile Tyr Ile Ser Lys Asp Phe Tyr Glu Thr 340 345 350 Leu Ser Cys Phe Met Ser Gly Asn Trp Asn Leu Ile Thr Gly Cys Val 355 360 365 Glu Asn Phe Tyr Asp Glu Asn Ile His Ala Lys Gly Lys Ser Lys Glu 370 375 380 Glu Lys Val Lys Lys Ala Val Lys Glu Asp Lys Tyr Lys Ser Ile Asn 385 390 395 400 Asp Val Asn Asp Leu Val Glu Lys Tyr Ile Asp Glu Lys Glu Arg Asn 405 410 415 Glu Phe Lys Asn Ser Asn Ala Lys Gln Tyr Ile Arg Glu Ile Ser Asn 420 425 430 Ile Ile Thr Asp Thr Glu Thr Ala His Leu Glu Tyr Asp Asp His Ile 435 440 445 Ser Leu Ile Glu Ser Glu Glu Lys Ala Asp Glu Met Lys Lys Arg Leu 450 455 460 Asp Met Tyr Met Asn Met Tyr His Trp Ala Lys Ala Phe Ile Val Asp 465 470 475 480 Glu Val Leu Asp Arg Asp Glu Met Phe Tyr Ser Asp Ile Asp Asp Ile 485 490 495 Tyr Asn Ile Leu Glu Asn Ile Val Pro Leu Tyr Asn Arg Val Arg Asn 500 505 510 Tyr Val Thr Gln Lys Pro Tyr Asn Ser Lys Lys Ile Lys Leu Asn Phe 515 520 525 Gln Ser Pro Thr Leu Ala Asn Gly Trp Ser Gln Ser Lys Glu Phe Asp 530 535 540 Asn Asn Ala Ile Ile Leu Ile Arg Asp Asn Lys Tyr Tyr Leu Ala Ile 545 550 555 560 Phe Asn Ala Lys Asn Lys Pro Asp Lys Lys Ile Ile Gln Gly Asn Ser 565 570 575 Asp Lys Lys Asn Asp Asn Asp Tyr Lys Lys Met Val Tyr Asn Leu Leu 580 585 590 Pro Gly Ala Asn Lys Met Leu Pro Lys Val Phe Leu Ser Lys Lys Gly 595 600 605 Ile Glu Thr Phe Lys Pro Ser Asp Tyr Ile Ile Ser Gly Tyr Asn Ala 610 615 620 His Lys His Ile Lys Thr Ser Glu Asn Phe Asp Ile Ser Phe Cys Arg 625 630 635 640 Asp Leu Ile Asp Tyr Phe Lys Asn Ser Ile Glu Lys His Ala Glu Trp 645 650 655 Arg Lys Tyr Glu Phe Lys Phe Ser Ala Thr Asp Ser Tyr Ser Asp Ile 660 665 670 Ser Glu Phe Tyr Arg Glu Val Glu Met Gln Gly Tyr Arg Ile Asp Trp 675 680 685 Thr Tyr Ile Ser Glu Ala Asp Ile Asn Lys Leu Asp Glu Glu Gly Lys 690 695 700 Ile Tyr Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ala Glu Asn Ser Thr 705 710 715 720 Gly Lys Glu Asn Leu His Thr Met Tyr Phe Lys Asn Ile Phe Ser Glu 725 730 735 Glu Asn Leu Asp Lys Ile Ile Lys Leu Asn Gly Gln Ala Glu Leu Phe 740 745 750 Tyr Arg Arg Ala Ser Val Lys Asn Pro Val Lys His Lys Lys Asp Ser 755 760 765 Val Leu Val Asn Lys Thr Tyr Lys Asn Gln Leu Asp Asn Gly Asp Val 770 775 780 Val Arg Ile Pro Ile Pro Asp Asp Ile Tyr Asn Glu Ile Tyr Lys Met 785 790 795 800 Tyr Asn Gly Tyr Ile Lys Glu Ser Asp Leu Ser Glu Ala Ala Lys Glu 805 810 815 Tyr Leu Asp Lys Val Glu Val Arg Thr Ala Gln Lys Asp Ile Val Lys 820 825 830 Asp Tyr Arg Tyr Thr Val Asp Lys Tyr Phe Ile His Thr Pro Ile Thr 835 840 845 Ile Asn Tyr Lys Val Thr Ala Arg Asn Asn Val Asn Asp Met Val Val 850 855 860 Lys Tyr Ile Ala Gln Asn Asp Asp Ile His Val Ile Gly Ile Asp Arg 865 870 875 880 Gly Glu Arg Asn Leu Ile Tyr Ile Ser Val Ile Asp Ser His Gly Asn 885 890 895 Ile Val Lys Gln Lys Ser Tyr Asn Ile Leu Asn Asn Tyr Asp Tyr Lys 900 905 910 Lys Lys Leu Val Glu Lys Glu Lys Thr Arg Glu Tyr Ala Arg Lys Asn 915 920 925 Trp Lys Ser Ile Gly Asn Ile Lys Glu Leu Lys Glu Gly Tyr Ile Ser 930 935 940 Gly Val Val His Glu Ile Ala Met Leu Ile Val Glu Tyr Asn Ala Ile 945 950 955 960 Ile Ala Met Glu Asp Leu Asn Tyr Gly Phe Lys Arg Gly Arg Phe Lys 965 970 975 Val Glu Arg Gln Val Tyr Gln Lys Phe Glu Ser Met Leu Ile Asn Lys 980 985 990 Leu Asn Tyr Phe Ala Ser Lys Glu Lys Ser Val Asp Glu Pro Gly Gly 995 1000 1005 Leu Leu Lys Gly Tyr Gln Leu Thr Tyr Val Pro Asp Asn Ile Lys 1010 1015 1020 Asn Leu Gly Lys Gln Cys Gly Val Ile Phe Tyr Val Pro Ala Ala 1025 1030 1035 Phe Thr Ser Lys Ile Asp Pro Ser Thr Gly Phe Ile Ser Ala Phe 1040 1045 1050 Asn Phe Lys Ser Ile Ser Thr Asn Ala Ser Arg Lys Gln Phe Phe 1055 1060 1065 Met Gln Phe Asp Glu Ile Arg Tyr Cys Ala Glu Lys Asp Met Phe 1070 1075 1080 Ser Phe Gly Phe Asp Tyr Asn Asn Phe Asp Thr Tyr Asn Ile Thr 1085 1090 1095 Met Gly Lys Thr Gln Trp Thr Val Tyr Thr Asn Gly Glu Arg Leu 1100 1105 1110 Gln Ser Glu Phe Asn Asn Ala Arg Arg Thr Gly Lys Thr Lys Ser 1115 1120 1125 Ile Asn Leu Thr Glu Thr Ile Lys Leu Leu Leu Glu Asp Asn Glu 1130 1135 1140 Ile Asn Tyr Ala Asp Gly His Asp Ile Arg Ile Asp Met Glu Lys 1145 1150 1155 Met Asp Glu Asp Lys Lys Ser Glu Phe Phe Ala Gln Leu Leu Ser 1160 1165 1170 Leu Tyr Lys Leu Thr Val Gln Met Arg Asn Ser Tyr Thr Glu Ala 1175 1180 1185 Glu Glu Gln Glu Asn Gly Ile Ser Tyr Asp Lys Ile Ile Ser Pro 1190 1195 1200 Val Ile Asn Asp Glu Gly Glu Phe Phe Asp Ser Asp Asn Tyr Lys 1205 1210 1215 Glu Ser Asp Asp Lys Glu Cys Lys Met Pro Lys Asp Ala Asp Ala 1220 1225 1230 Asn Gly Ala Tyr Cys Ile Ala Leu Lys Gly Leu Tyr Glu Val Leu 1235 1240 1245 Lys Ile Lys Ser Glu Trp Thr Glu Asp Gly Phe Asp Arg Asn Cys 1250 1255 1260 Leu Lys Leu Pro His Ala Glu Trp Leu Asp Phe Ile Gln Asn Lys 1265 1270 1275 Arg Tyr Glu 1280 <210> 34 <211> 1300 <212> PRT <213> Francisella novicida <400> 34 Met Ser Ile Tyr Gln Glu Phe Val Asn Lys Tyr Ser Leu Ser Lys Thr 1 5 10 15 Leu Arg Phe Glu Leu Ile Pro Gln Gly Lys Thr Leu Glu Asn Ile Lys 20 25 30 Ala Arg Gly Leu Ile Leu Asp Asp Glu Lys Arg Ala Lys Asp Tyr Lys 35 40 45 Lys Ala Lys Gln Ile Ile Asp Lys Tyr His Gln Phe Phe Ile Glu Glu 50 55 60 Ile Leu Ser Ser Val Cys Ile Ser Glu Asp Leu Leu Gln Asn Tyr Ser 65 70 75 80 Asp Val Tyr Phe Lys Leu Lys Lys Ser Asp Asp Asp Asn Leu Gln Lys 85 90 95 Asp Phe Lys Ser Ala Lys Asp Thr Ile Lys Lys Gln Ile Ser Glu Tyr 100 105 110 Ile Lys Asp Ser Glu Lys Phe Lys Asn Leu Phe Asn Gln Asn Leu Ile 115 120 125 Asp Ala Lys Lys Gly Gln Glu Ser Asp Leu Ile Leu Trp Leu Lys Gln 130 135 140 Ser Lys Asp Asn Gly Ile Glu Leu Phe Lys Ala Asn Ser Asp Ile Thr 145 150 155 160 Asp Ile Asp Glu Ala Leu Glu Ile Ile Lys Ser Phe Lys Gly Trp Thr 165 170 175 Thr Tyr Phe Lys Gly Phe His Glu Asn Arg Lys Val Asn Tyr Ser Ser 180 185 190 Asn Asp Ile Pro Thr Ser Ile Ile Tyr Arg Ile Val Asp Asp Asn Leu 195 200 205 Pro Lys Phe Leu Glu Asn Lys Ala Lys Tyr Glu Ser Leu Lys Asp Lys 210 215 220 Ala Pro Glu Ala Ile Asn Tyr Glu Gln Ile Lys Lys Asp Leu Ala Glu 225 230 235 240 Glu Leu Thr Phe Asp Ile Asp Tyr Lys Thr Ser Glu Val Asn Gln Arg 245 250 255 Val Phe Ser Leu Asp Glu Val Phe Glu Ile Ala Asn Phe Asn Asn Tyr 260 265 270 Leu Asn Gln Ser Gly Ile Thr Lys Phe Asn Thr Ile Ile Gly Gly Lys 275 280 285 Phe Val Asn Gly Glu Asn Thr Lys Arg Lys Gly Ile Asn Glu Tyr Ile 290 295 300 Asn Leu Tyr Ser Gln Gln Ile Asn Asp Lys Thr Leu Lys Lys Tyr Lys 305 310 315 320 Met Ser Val Leu Phe Lys Gln Ile Leu Ser Asp Thr Glu Ser Lys Ser 325 330 335 Phe Val Ile Asp Lys Leu Glu Asp Asp Ser Asp Val Val Thr Thr Met 340 345 350 Gln Ser Phe Tyr Glu Gln Ile Ala Ala Phe Lys Thr Val Glu Glu Lys 355 360 365 Ser Ile Lys Glu Thr Leu Ser Leu Leu Phe Asp Asp Leu Lys Ala Gln 370 375 380 Lys Leu Asp Leu Ser Lys Ile Tyr Phe Lys Asn Asp Lys Ser Leu Thr 385 390 395 400 Asp Leu Ser Gln Gln Val Phe Asp Asp Tyr Ser Val Ile Gly Thr Ala 405 410 415 Val Leu Glu Tyr Ile Thr Gln Gln Ile Ala Pro Lys Asn Leu Asp Asn 420 425 430 Pro Ser Lys Lys Glu Gln Glu Leu Ile Ala Lys Lys Thr Glu Lys Ala 435 440 445 Lys Tyr Leu Ser Leu Glu Thr Ile Lys Leu Ala Leu Glu Glu Phe Asn 450 455 460 Lys His Arg Asp Ile Asp Lys Gln Cys Arg Phe Glu Glu Ile Leu Ala 465 470 475 480 Asn Phe Ala Ala Ile Pro Met Ile Phe Asp Glu Ile Ala Gln Asn Lys 485 490 495 Asp Asn Leu Ala Gln Ile Ser Ile Lys Tyr Gln Asn Gln Gly Lys Lys 500 505 510 Asp Leu Leu Gln Ala Ser Ala Glu Asp Asp Val Lys Ala Ile Lys Asp 515 520 525 Leu Leu Asp Gln Thr Asn Asn Leu Leu His Lys Leu Lys Ile Phe His 530 535 540 Ile Ser Gln Ser Glu Asp Lys Ala Asn Ile Leu Asp Lys Asp Glu His 545 550 555 560 Phe Tyr Leu Val Phe Glu Glu Cys Tyr Phe Glu Leu Ala Asn Ile Val 565 570 575 Pro Leu Tyr Asn Lys Ile Arg Asn Tyr Ile Thr Gln Lys Pro Tyr Ser 580 585 590 Asp Glu Lys Phe Lys Leu Asn Phe Glu Asn Ser Thr Leu Ala Asn Gly 595 600 605 Trp Asp Lys Asn Lys Glu Pro Asp Asn Thr Ala Ile Leu Phe Ile Lys 610 615 620 Asp Asp Lys Tyr Tyr Leu Gly Val Met Asn Lys Lys Asn Asn Lys Ile 625 630 635 640 Phe Asp Asp Lys Ala Ile Lys Glu Asn Lys Gly Glu Gly Tyr Lys Lys 645 650 655 Ile Val Tyr Lys Leu Leu Pro Gly Ala Asn Lys Met Leu Pro Lys Val 660 665 670 Phe Phe Ser Ala Lys Ser Ile Lys Phe Tyr Asn Pro Ser Glu Asp Ile 675 680 685 Leu Arg Ile Arg Asn His Ser Thr His Thr Lys Asn Gly Ser Pro Gln 690 695 700 Lys Gly Tyr Glu Lys Phe Glu Phe Asn Ile Glu Asp Cys Arg Lys Phe 705 710 715 720 Ile Asp Phe Tyr Lys Gln Ser Ile Ser Lys His Pro Glu Trp Lys Asp 725 730 735 Phe Gly Phe Arg Phe Ser Asp Thr Gln Arg Tyr Asn Ser Ile Asp Glu 740 745 750 Phe Tyr Arg Glu Val Glu Asn Gln Gly Tyr Lys Leu Thr Phe Glu Asn 755 760 765 Ile Ser Glu Ser Tyr Ile Asp Ser Val Val Asn Gln Gly Lys Leu Tyr 770 775 780 Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ser Ala Tyr Ser Lys Gly Arg 785 790 795 800 Pro Asn Leu His Thr Leu Tyr Trp Lys Ala Leu Phe Asp Glu Arg Asn 805 810 815 Leu Gln Asp Val Val Tyr Lys Leu Asn Gly Glu Ala Glu Leu Phe Tyr 820 825 830 Arg Lys Gln Ser Ile Pro Lys Lys Ile Thr His Pro Ala Lys Glu Ala 835 840 845 Ile Ala Asn Lys Asn Lys Asp Asn Pro Lys Lys Glu Ser Val Phe Glu 850 855 860 Tyr Asp Leu Ile Lys Asp Lys Arg Phe Thr Glu Asp Lys Phe Phe Phe 865 870 875 880 His Cys Pro Ile Thr Ile Asn Phe Lys Ser Ser Gly Ala Asn Lys Phe 885 890 895 Asn Asp Glu Ile Asn Leu Leu Leu Lys Glu Lys Ala Asn Asp Val His 900 905 910 Ile Leu Ser Ile Asp Arg Gly Glu Arg His Leu Ala Tyr Tyr Thr Leu 915 920 925 Val Asp Gly Lys Gly Asn Ile Ile Lys Gln Asp Thr Phe Asn Ile Ile 930 935 940 Gly Asn Asp Arg Met Lys Thr Asn Tyr His Asp Lys Leu Ala Ala Ile 945 950 955 960 Glu Lys Asp Arg Asp Ser Ala Arg Lys Asp Trp Lys Lys Ile Asn Asn 965 970 975 Ile Lys Glu Met Lys Glu Gly Tyr Leu Ser Gln Val Val His Glu Ile 980 985 990 Ala Lys Leu Val Ile Glu Tyr Asn Ala Ile Val Val Phe Glu Asp Leu 995 1000 1005 Asn Phe Gly Phe Lys Arg Gly Arg Phe Lys Val Glu Lys Gln Val 1010 1015 1020 Tyr Gln Lys Leu Glu Lys Met Leu Ile Glu Lys Leu Asn Tyr Leu 1025 1030 1035 Val Phe Lys Asp Asn Glu Phe Asp Lys Thr Gly Gly Val Leu Arg 1040 1045 1050 Ala Tyr Gln Leu Thr Ala Pro Phe Glu Thr Phe Lys Lys Met Gly 1055 1060 1065 Lys Gln Thr Gly Ile Ile Tyr Tyr Val Pro Ala Gly Phe Thr Ser 1070 1075 1080 Lys Ile Cys Pro Val Thr Gly Phe Val Asn Gln Leu Tyr Pro Lys 1085 1090 1095 Tyr Glu Ser Val Ser Lys Ser Gln Glu Phe Phe Ser Lys Phe Asp 1100 1105 1110 Lys Ile Cys Tyr Asn Leu Asp Lys Gly Tyr Phe Glu Phe Ser Phe 1115 1120 1125 Asp Tyr Lys Asn Phe Gly Asp Lys Ala Ala Lys Gly Lys Trp Thr 1130 1135 1140 Ile Ala Ser Phe Gly Ser Arg Leu Ile Asn Phe Arg Asn Ser Asp 1145 1150 1155 Lys Asn His Asn Trp Asp Thr Arg Glu Val Tyr Pro Thr Lys Glu 1160 1165 1170 Leu Glu Lys Leu Leu Lys Asp Tyr Ser Ile Glu Tyr Gly His Gly 1175 1180 1185 Glu Cys Ile Lys Ala Ala Ile Cys Gly Glu Ser Asp Lys Lys Phe 1190 1195 1200 Phe Ala Lys Leu Thr Ser Val Leu Asn Thr Ile Leu Gln Met Arg 1205 1210 1215 Asn Ser Lys Thr Gly Thr Glu Leu Asp Tyr Leu Ile Ser Pro Val 1220 1225 1230 Ala Asp Val Asn Gly Asn Phe Phe Asp Ser Arg Gln Ala Pro Lys 1235 1240 1245 Asn Met Pro Gln Asp Ala Asp Ala Asn Gly Ala Tyr His Ile Gly 1250 1255 1260 Leu Lys Gly Leu Met Leu Leu Gly Arg Ile Lys Asn Asn Gln Glu 1265 1270 1275 Gly Lys Lys Leu Asn Leu Val Ile Lys Asn Glu Glu Tyr Phe Glu 1280 1285 1290 Phe Val Gln Asn Arg Asn Asn 1295 1300 <210> 35 <211> 1206 <212> PRT <213> Ruminococcaceae bacterium <400> 35 Met Tyr Tyr Glu Ser Leu Thr Lys Gln Tyr Pro Val Ser Lys Thr Ile 1 5 10 15 Arg Asn Glu Leu Ile Pro Ile Gly Lys Thr Leu Asp Asn Ile Arg Gln 20 25 30 Asn Asn Ile Leu Glu Ser Asp Val Lys Arg Lys Gln Asn Tyr Glu His 35 40 45 Val Lys Gly Ile Leu Asp Glu Tyr His Lys Gln Leu Ile Asn Glu Ala 50 55 60 Leu Asp Asn Cys Thr Leu Pro Ser Leu Lys Ile Ala Ala Glu Ile Tyr 65 70 75 80 Leu Lys Asn Gln Lys Glu Val Ser Asp Arg Glu Asp Phe Asn Lys Thr 85 90 95 Gln Asp Leu Leu Arg Lys Glu Val Val Glu Lys Leu Lys Ala His Glu 100 105 110 Asn Phe Thr Lys Ile Gly Lys Lys Asp Ile Leu Asp Leu Leu Glu Lys 115 120 125 Leu Pro Ser Ile Ser Glu Asp Asp Tyr Asn Ala Leu Glu Ser Phe Arg 130 135 140 Asn Phe Tyr Thr Tyr Phe Thr Ser Tyr Asn Lys Val Arg Glu Asn Leu 145 150 155 160 Tyr Ser Asp Lys Glu Lys Ser Ser Thr Val Ala Tyr Arg Leu Ile Asn 165 170 175 Glu Asn Phe Pro Lys Phe Leu Asp Asn Val Lys Ser Tyr Arg Phe Val 180 185 190 Lys Thr Ala Gly Ile Leu Ala Asp Gly Leu Gly Glu Glu Glu Gln Asp 195 200 205 Ser Leu Phe Ile Val Glu Thr Phe Asn Lys Thr Leu Thr Gln Asp Gly 210 215 220 Ile Asp Thr Tyr Asn Ser Gln Val Gly Lys Ile Asn Ser Ser Ile Asn 225 230 235 240 Leu Tyr Asn Gln Lys Asn Gln Lys Ala Asn Gly Phe Arg Lys Ile Pro 245 250 255 Lys Met Lys Met Leu Tyr Lys Gln Ile Leu Ser Asp Arg Glu Glu Ser 260 265 270 Phe Ile Asp Glu Phe Gln Ser Asp Glu Val Leu Ile Asp Asn Val Glu 275 280 285 Ser Tyr Gly Ser Val Leu Ile Glu Ser Leu Lys Ser Ser Lys Val Ser 290 295 300 Ala Phe Phe Asp Ala Leu Arg Glu Ser Lys Gly Lys Asn Val Tyr Val 305 310 315 320 Lys Asn Asp Leu Ala Lys Thr Ala Met Ser Val Ile Val Phe Glu Asn 325 330 335 Trp Arg Thr Phe Asp Asp Leu Leu Asn Gln Glu Tyr Asp Leu Ala Asn 340 345 350 Glu Asn Lys Lys Lys Asp Asp Lys Tyr Phe Glu Lys Arg Gln Lys Glu 355 360 365 Leu Lys Lys Asn Lys Ser Tyr Ser Leu Glu His Leu Cys Asn Leu Ser 370 375 380 Glu Asp Ser Cys Asn Leu Ile Glu Asn Tyr Ile His Gln Ile Ser Asp 385 390 395 400 Asp Ile Glu Asn Ile Ile Ile Asn Asn Glu Thr Phe Leu Arg Ile Val 405 410 415 Ile Asn Glu His Asp Arg Ser Arg Lys Leu Ala Lys Asn Arg Lys Ala 420 425 430 Val Lys Ala Ile Lys Asp Phe Leu Asp Ser Ile Lys Val Leu Glu Arg 435 440 445 Glu Leu Lys Leu Ile Asn Ser Ser Gly Gln Glu Leu Glu Lys Asp Leu 450 455 460 Ile Val Tyr Ser Ala His Glu Glu Leu Leu Val Glu Leu Lys Gln Val 465 470 475 480 Asp Ser Leu Tyr Asn Met Thr Arg Asn Tyr Leu Thr Lys Lys Pro Phe 485 490 495 Ser Thr Glu Lys Val Lys Leu Asn Phe Asn Arg Ser Thr Leu Leu Asn 500 505 510 Gly Trp Asp Arg Asn Lys Glu Thr Asp Asn Leu Gly Val Leu Leu Leu 515 520 525 Lys Asp Gly Lys Tyr Tyr Leu Gly Ile Met Asn Thr Ser Ala Asn Lys 530 535 540 Ala Phe Val Asn Pro Pro Val Ala Lys Thr Glu Lys Val Phe Lys Lys 545 550 555 560 Val Asp Tyr Lys Leu Leu Pro Val Pro Asn Gln Met Leu Pro Lys Val 565 570 575 Phe Phe Ala Lys Ser Asn Ile Asp Phe Tyr Asn Pro Ser Ser Glu Ile 580 585 590 Tyr Ser Asn Tyr Lys Lys Gly Thr His Lys Lys Gly Asn Met Phe Ser 595 600 605 Leu Glu Asp Cys His Asn Leu Ile Asp Phe Phe Lys Glu Ser Ile Ser 610 615 620 Lys His Glu Asp Trp Ser Lys Phe Gly Phe Lys Phe Asp Thr Gln Ala 625 630 635 640 Ser Tyr Asn Asp Ile Ser Glu Phe Tyr Arg Glu Val Glu Lys Gln Gly 645 650 655 Tyr Lys Leu Thr Tyr Thr Asp Ile Asp Glu Thr Tyr Ile Asn Asp Leu 660 665 670 Ile Glu Arg Asn Glu Leu Tyr Leu Phe Gln Ile Tyr Asn Lys Asp Phe 675 680 685 Ser Met Tyr Ser Lys Gly Lys Leu Asn Leu His Thr Leu Tyr Phe Met 690 695 700 Met Leu Phe Asp Gln Arg Asn Ile Asp Asp Val Val Tyr Lys Leu Asn 705 710 715 720 Gly Glu Ala Glu Val Phe Tyr Arg Pro Ala Ser Ile Ser Glu Asp Glu 725 730 735 Leu Ile Ile His Lys Ala Gly Glu Glu Ile Lys Asn Lys Asn Pro Asn 740 745 750 Arg Ala Arg Thr Lys Glu Thr Ser Thr Phe Ser Tyr Asp Ile Val Lys 755 760 765 Asp Lys Arg Tyr Ser Lys Asp Lys Phe Thr Leu His Ile Pro Ile Thr 770 775 780 Met Asn Phe Gly Val Asp Glu Val Lys Arg Phe Asn Asp Ala Val Asn 785 790 795 800 Ser Ala Ile Arg Ile Asp Glu Asn Val Asn Val Ile Gly Ile Asp Arg 805 810 815 Gly Glu Arg Asn Leu Leu Tyr Val Val Val Ile Asp Ser Lys Gly Asn 820 825 830 Ile Leu Glu Gln Ile Ser Leu Asn Ser Ile Ile Asn Lys Glu Tyr Asp 835 840 845 Ile Glu Thr Asp Tyr His Ala Leu Leu Asp Glu Arg Glu Gly Gly Arg 850 855 860 Asp Lys Ala Arg Lys Asp Trp Asn Thr Val Glu Asn Ile Arg Asp Leu 865 870 875 880 Lys Ala Gly Leu Tyr Leu Gln Val Val Asn Val Val Ala Lys Leu Val 885 890 895 Leu Lys Tyr Asn Ala Ile Ile Cys Leu Glu Asp Leu Asn Phe Gly Phe 900 905 910 Lys Arg Gly Arg Gln Lys Val Glu Lys Gln Val Tyr Gln Lys Phe Glu 915 920 925 Lys Met Leu Ile Asp Lys Leu Asn Tyr Leu Val Ile Asp Lys Ser Arg 930 935 940 Glu Gln Thr Ser Pro Lys Glu Leu Gly Gly Ala Leu Asn Ala Leu Gln 945 950 955 960 Leu Thr Ser Lys Phe Lys Ser Phe Lys Glu Leu Gly Lys Gln Ser Gly 965 970 975 Val Ile Tyr Tyr Val Pro Ala Tyr Leu Thr Ser Lys Ile Asp Pro Thr 980 985 990 Thr Gly Phe Ala Asn Leu Phe Tyr Met Lys Cys Glu Asn Val Glu Lys 995 1000 1005 Ser Lys Arg Phe Phe Asp Gly Phe Asp Phe Ile Arg Phe Asn Ala 1010 1015 1020 Leu Glu Asn Val Phe Glu Phe Gly Phe Asp Tyr Arg Ser Phe Thr 1025 1030 1035 Gln Arg Ala Cys Gly Ile Asn Ser Lys Trp Thr Val Cys Thr Asn 1040 1045 1050 Gly Glu Arg Ile Ile Lys Tyr Arg Asn Pro Asp Lys Asn Asn Met 1055 1060 1065 Phe Asp Glu Lys Val Val Val Val Thr Asp Glu Met Lys Asn Leu 1070 1075 1080 Phe Glu Gln Tyr Lys Ile Pro Tyr Glu Asp Gly Arg Asn Val Lys 1085 1090 1095 Asp Met Ile Ile Ser Asn Glu Glu Ala Glu Phe Tyr Arg Arg Leu 1100 1105 1110 Tyr Arg Leu Leu Gln Gln Thr Leu Gln Met Arg Asn Ser Thr Ser 1115 1120 1125 Asp Gly Thr Arg Asp Tyr Ile Ile Ser Pro Val Lys Asn Lys Arg 1130 1135 1140 Glu Ala Tyr Phe Asn Ser Glu Leu Ser Asp Gly Ser Val Pro Lys 1145 1150 1155 Asp Ala Asp Ala Asn Gly Ala Tyr Asn Ile Ala Arg Lys Gly Leu 1160 1165 1170 Trp Val Leu Glu Gln Ile Arg Gln Lys Ser Glu Gly Glu Lys Ile 1175 1180 1185 Asn Leu Ala Met Thr Asn Ala Glu Trp Leu Glu Tyr Ala Gln Thr 1190 1195 1200 His Leu Leu 1205 <210> 36 <211> 1233 <212> PRT <213> Lachnospiraceae bacterium <400> 36 Met Asp Tyr Gly Asn Gly Gln Phe Glu Arg Arg Ala Pro Leu Thr Lys 1 5 10 15 Thr Ile Thr Leu Arg Leu Lys Pro Ile Gly Glu Thr Arg Glu Thr Ile 20 25 30 Arg Glu Gln Lys Leu Leu Glu Gln Asp Ala Ala Phe Arg Lys Leu Val 35 40 45 Glu Thr Val Thr Pro Ile Val Asp Asp Cys Ile Arg Lys Ile Ala Asp 50 55 60 Asn Ala Leu Cys His Phe Gly Thr Glu Tyr Asp Phe Ser Cys Leu Gly 65 70 75 80 Asn Ala Ile Ser Lys Asn Asp Ser Lys Ala Ile Lys Lys Glu Thr Glu 85 90 95 Lys Val Glu Lys Leu Leu Ala Lys Val Leu Thr Glu Asn Leu Pro Asp 100 105 110 Gly Leu Arg Lys Val Asn Asp Ile Asn Ser Ala Ala Phe Ile Gln Asp 115 120 125 Thr Leu Thr Ser Phe Val Gln Asp Asp Ala Asp Lys Arg Val Leu Ile 130 135 140 Gln Glu Leu Lys Gly Lys Thr Val Leu Met Gln Arg Phe Leu Thr Thr 145 150 155 160 Arg Ile Thr Ala Leu Thr Val Trp Leu Pro Asp Arg Val Phe Glu Asn 165 170 175 Phe Asn Ile Phe Ile Glu Asn Ala Glu Lys Met Arg Ile Leu Leu Asp 180 185 190 Ser Pro Leu Asn Glu Lys Ile Met Lys Phe Asp Pro Asp Ala Glu Gln 195 200 205 Tyr Ala Ser Leu Glu Phe Tyr Gly Gln Cys Leu Ser Gln Lys Asp Ile 210 215 220 Asp Ser Tyr Asn Leu Ile Ile Ser Gly Ile Tyr Ala Asp Asp Glu Val 225 230 235 240 Lys Asn Pro Gly Ile Asn Glu Ile Val Lys Glu Tyr Asn Gln Gln Ile 245 250 255 Arg Gly Asp Lys Asp Glu Ser Pro Leu Pro Lys Leu Lys Lys Leu His 260 265 270 Lys Gln Ile Leu Met Pro Val Glu Lys Ala Phe Phe Val Arg Val Leu 275 280 285 Ser Asn Asp Ser Asp Ala Arg Ser Ile Leu Glu Lys Ile Leu Lys Asp 290 295 300 Thr Glu Met Leu Pro Ser Lys Ile Ile Glu Ala Met Lys Glu Ala Asp 305 310 315 320 Ala Gly Asp Ile Ala Val Tyr Gly Ser Arg Leu His Glu Leu Ser His 325 330 335 Val Ile Tyr Gly Asp His Gly Lys Leu Ser Gln Ile Ile Tyr Asp Lys 340 345 350 Glu Ser Lys Arg Ile Ser Glu Leu Met Glu Thr Leu Ser Pro Lys Glu 355 360 365 Arg Lys Glu Ser Lys Lys Arg Leu Glu Gly Leu Glu Glu His Ile Arg 370 375 380 Lys Ser Thr Tyr Thr Phe Asp Glu Leu Asn Arg Tyr Ala Glu Lys Asn 385 390 395 400 Val Met Ala Ala Tyr Ile Ala Ala Val Glu Glu Ser Cys Ala Glu Ile 405 410 415 Met Arg Lys Glu Lys Asp Leu Arg Thr Leu Leu Ser Lys Glu Asp Val 420 425 430 Lys Ile Arg Gly Asn Arg His Asn Thr Leu Ile Val Lys Asn Tyr Phe 435 440 445 Asn Ala Trp Thr Val Phe Arg Asn Leu Ile Arg Ile Leu Arg Arg Lys 450 455 460 Ser Glu Ala Glu Ile Asp Ser Asp Phe Tyr Asp Val Leu Asp Asp Ser 465 470 475 480 Val Glu Val Leu Ser Leu Thr Tyr Lys Gly Glu Asn Leu Cys Arg Ser 485 490 495 Tyr Ile Thr Lys Lys Ile Gly Ser Asp Leu Lys Pro Glu Ile Ala Thr 500 505 510 Tyr Gly Ser Ala Leu Arg Pro Asn Ser Arg Trp Trp Ser Pro Gly Glu 515 520 525 Lys Phe Asn Val Lys Phe His Thr Ile Val Arg Arg Asp Gly Arg Leu 530 535 540 Tyr Tyr Phe Ile Leu Pro Lys Gly Ala Lys Pro Val Glu Leu Glu Asp 545 550 555 560 Met Asp Gly Asp Ile Glu Cys Leu Gln Met Arg Lys Ile Pro Asn Pro 565 570 575 Thr Ile Phe Leu Pro Lys Leu Val Phe Lys Asp Pro Glu Ala Phe Phe 580 585 590 Arg Asp Asn Pro Glu Ala Asp Glu Phe Val Phe Leu Ser Gly Met Lys 595 600 605 Ala Pro Val Thr Ile Thr Arg Glu Thr Tyr Glu Ala Tyr Arg Tyr Lys 610 615 620 Leu Tyr Thr Val Gly Lys Leu Arg Asp Gly Glu Val Ser Glu Glu Glu 625 630 635 640 Tyr Lys Arg Ala Leu Leu Gln Val Leu Thr Ala Tyr Lys Glu Phe Leu 645 650 655 Glu Asn Arg Met Ile Tyr Ala Asp Leu Asn Phe Gly Phe Lys Asp Leu 660 665 670 Glu Glu Tyr Lys Asp Ser Ser Glu Phe Ile Lys Gln Val Glu Thr His 675 680 685 Asn Thr Phe Met Cys Trp Ala Lys Val Ser Ser Ser Gln Leu Asp Asp 690 695 700 Leu Val Lys Ser Gly Asn Gly Leu Leu Phe Glu Ile Trp Ser Glu Arg 705 710 715 720 Leu Glu Ser Tyr Tyr Lys Tyr Gly Asn Glu Lys Val Leu Arg Gly Tyr 725 730 735 Glu Gly Val Leu Leu Ser Ile Leu Lys Asp Glu Asn Leu Val Ser Met 740 745 750 Arg Thr Leu Leu Asn Ser Arg Pro Met Leu Val Tyr Arg Pro Lys Glu 755 760 765 Ser Ser Lys Pro Met Val Val His Arg Asp Gly Ser Arg Val Val Asp 770 775 780 Arg Phe Asp Lys Asp Gly Lys Tyr Ile Pro Pro Glu Val His Asp Glu 785 790 795 800 Leu Tyr Arg Phe Phe Asn Asn Leu Leu Ile Lys Glu Lys Leu Gly Glu 805 810 815 Lys Ala Arg Lys Ile Leu Asp Asn Lys Lys Val Lys Val Lys Val Leu 820 825 830 Glu Ser Glu Arg Val Lys Trp Ser Lys Phe Tyr Asp Glu Gln Phe Ala 835 840 845 Val Thr Phe Ser Val Lys Lys Asn Ala Asp Cys Leu Asp Thr Thr Lys 850 855 860 Asp Leu Asn Ala Glu Val Met Glu Gln Tyr Ser Glu Ser Asn Arg Leu 865 870 875 880 Ile Leu Ile Arg Asn Thr Thr Asp Ile Leu Tyr Tyr Leu Val Leu Asp 885 890 895 Lys Asn Gly Lys Val Leu Lys Gln Arg Ser Leu Asn Ile Ile Asn Asp 900 905 910 Gly Ala Arg Asp Val Asp Trp Lys Glu Arg Phe Arg Gln Val Thr Lys 915 920 925 Asp Arg Asn Glu Gly Tyr Asn Glu Trp Asp Tyr Ser Arg Thr Ser Asn 930 935 940 Asp Leu Lys Glu Val Tyr Leu Asn Tyr Ala Leu Lys Glu Ile Ala Glu 945 950 955 960 Ala Val Ile Glu Tyr Asn Ala Ile Leu Ile Ile Glu Lys Met Ser Asn 965 970 975 Ala Phe Lys Asp Lys Tyr Ser Phe Leu Asp Asp Val Thr Phe Lys Gly 980 985 990 Phe Glu Thr Lys Lys Leu Ala Lys Leu Ser Asp Leu His Phe Arg Gly 995 1000 1005 Ile Lys Asp Gly Glu Pro Cys Ser Phe Thr Asn Pro Leu Gln Leu 1010 1015 1020 Cys Gln Asn Asp Ser Asn Lys Ile Leu Gln Asp Gly Val Ile Phe 1025 1030 1035 Met Val Pro Asn Ser Met Thr Arg Ser Leu Asp Pro Asp Thr Gly 1040 1045 1050 Phe Ile Phe Ala Ile Asn Asp His Asn Ile Arg Thr Lys Lys Ala 1055 1060 1065 Lys Leu Asn Phe Leu Ser Lys Phe Asp Gln Leu Lys Val Ser Ser 1070 1075 1080 Glu Gly Cys Leu Ile Met Lys Tyr Ser Gly Asp Ser Leu Pro Thr 1085 1090 1095 His Asn Thr Asp Asn Arg Val Trp Asn Cys Cys Cys Asn His Pro 1100 1105 1110 Ile Thr Asn Tyr Asp Arg Glu Thr Lys Lys Val Glu Phe Ile Glu 1115 1120 1125 Glu Pro Val Glu Glu Leu Ser Arg Val Leu Glu Glu Asn Gly Ile 1130 1135 1140 Glu Thr Asp Thr Glu Leu Asn Lys Leu Asn Glu Arg Glu Asn Val 1145 1150 1155 Pro Gly Lys Val Val Asp Ala Ile Tyr Ser Leu Val Leu Asn Tyr 1160 1165 1170 Leu Arg Gly Thr Val Ser Gly Val Ala Gly Gln Arg Ala Val Tyr 1175 1180 1185 Tyr Ser Pro Val Thr Gly Lys Lys Tyr Asp Ile Ser Phe Ile Gln 1190 1195 1200 Ala Met Asn Leu Asn Arg Lys Cys Asp Tyr Tyr Arg Ile Gly Ser 1205 1210 1215 Lys Glu Arg Gly Glu Trp Thr Asp Phe Val Ala Gln Leu Ile Asn 1220 1225 1230 <210> 37 <211> 1227 <212> PRT <213> Ruminococcaceae bacterium <400> 37 Met Ser Lys Leu Glu Lys Phe Thr Asn Cys Tyr Ser Leu Ser Lys Thr 1 5 10 15 Leu Arg Phe Lys Ala Ile Pro Val Gly Lys Thr Gln Glu Asn Ile Asp 20 25 30 Asn Lys Arg Leu Leu Val Glu Asp Glu Lys Arg Ala Glu Asp Tyr Lys 35 40 45 Gly Val Lys Lys Leu Leu Asp Arg Tyr Tyr Leu Ser Phe Ile Asn Asp 50 55 60 Val Leu His Ser Ile Lys Leu Lys Asn Leu Asn Asn Tyr Ile Ser Leu 65 70 75 80 Phe Arg Lys Lys Thr Arg Thr Glu Lys Glu Asn Lys Glu Leu Glu Asn 85 90 95 Leu Glu Ile Asn Leu Arg Lys Glu Ile Ala Lys Ala Phe Lys Gly Asn 100 105 110 Glu Gly Tyr Lys Ser Leu Phe Lys Lys Asp Ile Ile Glu Thr Ile Leu 115 120 125 Pro Glu Phe Leu Asp Asp Lys Asp Glu Ile Ala Leu Val Asn Ser Phe 130 135 140 Asn Gly Phe Thr Thr Ala Phe Thr Gly Phe Phe Asp Asn Arg Glu Asn 145 150 155 160 Met Phe Ser Glu Glu Ala Lys Ser Thr Ser Ile Ala Phe Arg Cys Ile 165 170 175 Asn Glu Asn Leu Thr Arg Tyr Ile Ser Asn Met Asp Ile Phe Glu Lys 180 185 190 Val Asp Ala Ile Phe Asp Lys His Glu Val Gln Glu Ile Lys Glu Lys 195 200 205 Ile Leu Asn Ser Asp Tyr Asp Val Glu Asp Phe Phe Glu Gly Glu Phe 210 215 220 Phe Asn Phe Val Leu Thr Gln Glu Gly Ile Asp Val Tyr Asn Ala Ile 225 230 235 240 Ile Gly Gly Phe Val Thr Glu Ser Gly Glu Lys Ile Lys Gly Leu Asn 245 250 255 Glu Tyr Ile Asn Leu Tyr Asn Gln Lys Thr Lys Gln Lys Leu Pro Lys 260 265 270 Phe Lys Pro Leu Tyr Lys Gln Val Leu Ser Asp Arg Glu Ser Leu Ser 275 280 285 Phe Tyr Gly Glu Gly Tyr Thr Ser Asp Glu Glu Val Leu Glu Val Phe 290 295 300 Arg Asn Thr Leu Asn Lys Asn Ser Glu Ile Phe Ser Ser Ile Lys Lys 305 310 315 320 Leu Glu Lys Leu Phe Lys Asn Phe Asp Glu Tyr Ser Ser Ala Gly Ile 325 330 335 Phe Val Lys Asn Gly Pro Ala Ile Ser Thr Ile Ser Lys Asp Ile Phe 340 345 350 Gly Glu Trp Asn Val Ile Arg Asp Lys Trp Asn Ala Glu Tyr Asp Asp 355 360 365 Ile His Leu Lys Lys Lys Ala Val Val Thr Glu Lys Tyr Glu Asp Asp 370 375 380 Arg Arg Lys Ser Phe Lys Lys Ile Gly Ser Phe Ser Leu Glu Gln Leu 385 390 395 400 Gln Glu Tyr Ala Asp Ala Asp Leu Ser Val Val Glu Lys Leu Lys Glu 405 410 415 Ile Ile Ile Gln Lys Val Asp Glu Ile Tyr Lys Val Tyr Gly Ser Ser 420 425 430 Glu Lys Leu Phe Asp Ala Asp Phe Val Leu Glu Lys Ser Leu Lys Lys 435 440 445 Asn Asp Ala Val Val Ala Ile Met Lys Asp Leu Leu Asp Ser Val Lys 450 455 460 Ser Phe Glu Asn Tyr Ile Lys Ala Phe Phe Gly Glu Gly Lys Glu Thr 465 470 475 480 Asn Arg Asp Glu Ser Phe Tyr Gly Asp Phe Val Leu Ala Tyr Asp Ile 485 490 495 Leu Leu Lys Val Asp His Ile Tyr Asp Ala Ile Arg Asn Tyr Val Thr 500 505 510 Gln Lys Pro Tyr Ser Lys Asp Lys Phe Lys Leu Tyr Phe Gln Asn Pro 515 520 525 Gln Phe Met Gly Gly Trp Asp Lys Asp Lys Glu Thr Asp Tyr Arg Ala 530 535 540 Thr Ile Leu Arg Tyr Gly Ser Lys Tyr Tyr Leu Ala Ile Met Asp Lys 545 550 555 560 Lys Tyr Ala Lys Cys Leu Gln Lys Ile Asp Lys Asp Asp Val Asn Gly 565 570 575 Asn Tyr Glu Lys Ile Asn Tyr Lys Leu Leu Pro Gly Pro Asn Lys Met 580 585 590 Leu Pro Lys Val Phe Phe Ser Lys Lys Trp Met Ala Tyr Tyr Asn Pro 595 600 605 Ser Glu Asp Ile Gln Lys Ile Tyr Lys Asn Gly Thr Phe Lys Lys Gly 610 615 620 Asp Met Phe Asn Leu Asn Asp Cys His Lys Leu Ile Asp Phe Phe Lys 625 630 635 640 Asp Ser Ile Ser Arg Tyr Pro Lys Trp Ser Asn Ala Tyr Asp Phe Asn 645 650 655 Phe Ser Glu Thr Glu Lys Tyr Lys Asp Ile Ala Gly Phe Tyr Arg Glu 660 665 670 Val Glu Glu Gln Gly Tyr Lys Val Ser Phe Glu Ser Ala Ser Lys Lys 675 680 685 Glu Val Asp Lys Leu Val Glu Glu Gly Lys Leu Tyr Met Phe Gln Ile 690 695 700 Tyr Asn Lys Asp Phe Ser Asp Lys Ser His Gly Thr Pro Asn Leu His 705 710 715 720 Thr Met Tyr Phe Lys Leu Leu Phe Asp Glu Asn Asn His Gly Gln Ile 725 730 735 Arg Leu Ser Gly Gly Ala Glu Leu Phe Met Arg Arg Ala Ser Leu Lys 740 745 750 Lys Glu Glu Leu Val Val His Pro Ala Asn Ser Pro Ile Ala Asn Lys 755 760 765 Asn Pro Asp Asn Pro Lys Lys Thr Thr Thr Leu Ser Tyr Asp Val Tyr 770 775 780 Lys Asp Lys Arg Phe Ser Glu Asp Gln Tyr Glu Leu His Ile Pro Ile 785 790 795 800 Ala Asn Ile Asn Lys Cys Pro Lys Asn Ile Phe Lys Ile Asn Thr Glu 805 810 815 Val Arg Val Leu Leu Lys His Asp Asp Asn Pro Tyr Val Ile Gly Ile 820 825 830 Asp Arg Gly Glu Arg Asn Leu Leu Tyr Ile Val Val Val Asp Gly Lys 835 840 845 Gly Asn Ile Val Glu Gln Tyr Ser Leu Asn Glu Ile Ile Asn Asn Phe 850 855 860 Asn Gly Ile Arg Ile Lys Thr Asp Tyr His Ser Leu Leu Asp Lys Lys 865 870 875 880 Glu Lys Glu Arg Phe Glu Ala Arg Gln Asn Trp Thr Ser Ile Glu Asn 885 890 895 Ile Lys Glu Leu Lys Ala Gly Tyr Ile Ser Gln Val Val His Lys Ile 900 905 910 Cys Glu Leu Val Glu Lys Tyr Asp Ala Val Ile Ala Leu Glu Asp Leu 915 920 925 Asn Ser Gly Phe Lys Asn Ser Arg Val Lys Val Glu Lys Gln Val Tyr 930 935 940 Gln Lys Phe Glu Lys Met Leu Ile Asp Lys Leu Asn Tyr Met Val Asp 945 950 955 960 Lys Lys Ser Asn Pro Cys Ala Thr Gly Gly Ala Leu Lys Gly Tyr Gln 965 970 975 Ile Thr Asn Lys Phe Glu Ser Phe Lys Ser Met Ser Thr Gln Asn Gly 980 985 990 Phe Ile Phe Tyr Ile Pro Ala Trp Leu Thr Ser Lys Ile Asp Pro Ser 995 1000 1005 Thr Gly Phe Val Asn Leu Leu Lys Thr Lys Tyr Thr Ser Ile Ala 1010 1015 1020 Asp Lys Lys Phe Ile Ser Ser Phe Asp Arg Ile Met Tyr Val Pro 1025 1030 1035 Glu Glu Asp Leu Phe Glu Phe Ala Leu Asp Tyr Lys Asn Phe Ser 1040 1045 1050 Arg Thr Asp Ala Asp Tyr Ile Lys Lys Trp Lys Leu Tyr Ser Tyr 1055 1060 1065 Gly Asn Arg Ile Arg Ile Phe Arg Asn Pro Lys Lys Asn Asn Val 1070 1075 1080 Phe Asp Trp Glu Glu Val Cys Leu Thr Ser Ala Tyr Lys Glu Leu 1085 1090 1095 Phe Asn Lys Tyr Gly Ile Asn Tyr Gln Gln Gly Asp Ile Arg Ala 1100 1105 1110 Leu Leu Cys Glu Gln Ser Asp Lys Ala Phe Tyr Ser Ser Phe Met 1115 1120 1125 Ala Leu Met Ser Leu Met Leu Gln Met Arg Asn Ser Ile Thr Gly 1130 1135 1140 Arg Thr Asp Val Asp Phe Leu Ile Ser Pro Val Lys Asn Ser Asp 1145 1150 1155 Gly Ile Phe Tyr Asp Ser Arg Asn Tyr Glu Ala Gln Glu Asn Ala 1160 1165 1170 Ile Leu Pro Lys Asn Ala Asp Ala Asn Gly Ala Tyr Asn Ile Ala 1175 1180 1185 Arg Lys Val Leu Trp Ala Ile Gly Gln Phe Lys Lys Ala Glu Asp 1190 1195 1200 Glu Lys Leu Asp Lys Val Lys Ile Ala Ser Asn Lys Glu Trp Leu 1205 1210 1215 Glu Tyr Ala Gln Thr Ser Val Lys His 1220 1225 <210> 38 <211> 1264 <212> PRT <213> Ruminococcaceae bacterium <400> 38 Met Glu Asp Tyr Ser Gly Phe Val Asn Ile Tyr Ser Ile Gln Lys Thr 1 5 10 15 Leu Arg Phe Glu Leu Lys Pro Val Gly Lys Thr Leu Glu His Ile Glu 20 25 30 Lys Lys Gly Phe Leu Lys Lys Asp Lys Ile Arg Ala Glu Asp Tyr Lys 35 40 45 Ala Val Lys Lys Ile Ile Asp Lys Tyr His Arg Ala Tyr Ile Glu Glu 50 55 60 Val Phe Asp Ser Val Leu His Gln Lys Lys Lys Lys Asp Lys Thr Arg 65 70 75 80 Phe Ser Thr Gln Phe Ile Lys Glu Ile Lys Glu Phe Ser Glu Leu Tyr 85 90 95 Tyr Lys Thr Glu Lys Asn Ile Pro Asp Lys Glu Arg Leu Glu Ala Leu 100 105 110 Ser Glu Lys Leu Arg Lys Met Leu Val Gly Ala Phe Lys Gly Glu Phe 115 120 125 Ser Glu Glu Val Ala Glu Lys Tyr Asn Lys Asn Leu Phe Ser Lys Glu 130 135 140 Leu Ile Arg Asn Glu Ile Glu Lys Phe Cys Glu Thr Asp Glu Glu Arg 145 150 155 160 Lys Gln Val Ser Asn Phe Lys Ser Phe Thr Thr Tyr Phe Thr Gly Phe 165 170 175 His Ser Asn Arg Gln Asn Ile Tyr Ser Asp Glu Lys Lys Ser Thr Ala 180 185 190 Ile Gly Tyr Arg Ile Ile His Gln Asn Leu Pro Lys Phe Leu Asp Asn 195 200 205 Leu Lys Ile Ile Glu Ser Ile Gln Arg Arg Phe Lys Asp Phe Pro Trp 210 215 220 Ser Asp Leu Lys Lys Asn Leu Lys Lys Ile Asp Lys Asn Ile Lys Leu 225 230 235 240 Thr Glu Tyr Phe Ser Ile Asp Gly Phe Val Asn Val Leu Asn Gln Lys 245 250 255 Gly Ile Asp Ala Tyr Asn Thr Ile Leu Gly Gly Lys Ser Glu Glu Ser 260 265 270 Gly Glu Lys Ile Gln Gly Leu Asn Glu Tyr Ile Asn Leu Tyr Arg Gln 275 280 285 Lys Asn Asn Ile Asp Arg Lys Asn Pro Leu Asn Val Lys Ile Leu Phe 290 295 300 Lys Gln Ile Leu Gly Asp Arg Glu Thr Lys Ser Phe Ile Pro Glu Ala 305 310 315 320 Phe Pro Asp Asp Gln Ser Val Leu Asn Ser Ile Thr Glu Phe Ala Lys 325 330 335 Tyr Leu Lys Leu Asp Lys Lys Lys Lys Ser Ile Ile Ala Glu Leu Lys 340 345 350 Lys Phe Leu Ser Ser Phe Asn Arg Tyr Glu Leu Asp Gly Ile Tyr Leu 355 360 365 Ala Asn Asp Asn Ser Leu Ala Ser Ile Ser Thr Phe Leu Phe Asp Asp 370 375 380 Trp Ser Phe Ile Lys Lys Ser Val Ser Phe Lys Tyr Asp Glu Ser Val 385 390 395 400 Gly Asp Pro Lys Lys Lys Ile Lys Ser Pro Leu Lys Tyr Glu Lys Glu 405 410 415 Lys Glu Lys Trp Leu Lys Gln Lys Tyr Tyr Thr Ile Ser Phe Leu Asn 420 425 430 Asp Ala Ile Glu Ser Tyr Ser Lys Ser Gln Asp Glu Lys Arg Val Lys 435 440 445 Ile Arg Leu Glu Ala Tyr Phe Ala Glu Phe Lys Ser Lys Asp Asp Ala 450 455 460 Lys Lys Gln Phe Asp Leu Leu Glu Arg Ile Glu Glu Ala Tyr Ala Ile 465 470 475 480 Val Glu Pro Leu Leu Gly Ala Glu Tyr Pro Arg Asp Arg Asn Leu Lys 485 490 495 Ala Asp Lys Lys Glu Val Gly Lys Ile Lys Asp Phe Leu Asp Ser Ile 500 505 510 Lys Ser Leu Gln Phe Phe Leu Lys Pro Leu Leu Ser Ala Glu Ile Phe 515 520 525 Asp Glu Lys Asp Leu Gly Phe Tyr Asn Gln Leu Glu Gly Tyr Tyr Glu 530 535 540 Glu Ile Asp Ile Ser Gly His Leu Tyr Asn Lys Val Arg Asn Tyr Leu 545 550 555 560 Thr Gly Lys Ile Tyr Ser Lys Glu Lys Phe Lys Leu Asn Phe Glu Asn 565 570 575 Ser Thr Leu Leu Lys Gly Trp Asp Glu Asn Arg Glu Val Ala Asn Leu 580 585 590 Cys Val Ile Phe Arg Glu Asp Gln Lys Tyr Tyr Leu Gly Val Met Asp 595 600 605 Lys Glu Asn Asn Thr Ile Leu Ser Asp Ile Pro Lys Val Lys Pro Asn 610 615 620 Glu Leu Phe Tyr Glu Lys Met Val Tyr Lys Leu Ile Pro Thr Pro His 625 630 635 640 Met Gln Leu Pro Arg Ile Ile Phe Ser Ser Asp Asn Leu Ser Ile Tyr 645 650 655 Asn Pro Ser Lys Ser Ile Leu Lys Ile Arg Glu Ala Lys Ser Phe Lys 660 665 670 Glu Gly Lys Asn Phe Lys Leu Lys Asp Cys His Lys Phe Ile Asp Phe 675 680 685 Tyr Lys Glu Ser Ile Ser Lys Asn Glu Asp Trp Ser Arg Phe Asp Phe 690 695 700 Lys Phe Ser Lys Thr Ser Ser Tyr Glu Asn Ile Ser Glu Phe Tyr Arg 705 710 715 720 Glu Val Glu Arg Gln Gly Tyr Asn Leu Asp Phe Lys Lys Val Ser Lys 725 730 735 Phe Tyr Ile Asp Ser Leu Val Glu Asp Gly Lys Leu Tyr Leu Phe Gln 740 745 750 Ile Tyr Asn Lys Asp Phe Ser Ile Phe Ser Lys Gly Lys Pro Asn Leu 755 760 765 His Thr Ile Tyr Phe Arg Ser Leu Phe Ser Lys Glu Asn Leu Lys Asp 770 775 780 Val Cys Leu Lys Leu Asn Gly Glu Ala Glu Met Phe Phe Arg Lys Lys 785 790 795 800 Ser Ile Asn Tyr Asp Glu Lys Lys Lys Arg Glu Gly His His Pro Glu 805 810 815 Leu Phe Glu Lys Leu Lys Tyr Pro Ile Leu Lys Asp Lys Arg Tyr Ser 820 825 830 Glu Asp Lys Phe Gln Phe His Leu Pro Ile Ser Leu Asn Phe Lys Ser 835 840 845 Lys Glu Arg Leu Asn Phe Asn Leu Lys Val Asn Glu Phe Leu Lys Arg 850 855 860 Asn Lys Asp Ile Asn Ile Ile Gly Ile Asp Arg Gly Glu Arg Asn Leu 865 870 875 880 Leu Tyr Leu Val Met Ile Asn Gln Lys Gly Glu Ile Leu Lys Gln Thr 885 890 895 Leu Leu Asp Ser Met Gln Ser Gly Lys Gly Arg Pro Glu Ile Asn Tyr 900 905 910 Lys Glu Lys Leu Gln Glu Lys Glu Ile Glu Arg Asp Lys Ala Arg Lys 915 920 925 Ser Trp Gly Thr Val Glu Asn Ile Lys Glu Leu Lys Glu Gly Tyr Leu 930 935 940 Ser Ile Val Ile His Gln Ile Ser Lys Leu Met Val Glu Asn Asn Ala 945 950 955 960 Ile Val Val Leu Glu Asp Leu Asn Ile Gly Phe Lys Arg Gly Arg Gln 965 970 975 Lys Val Glu Arg Gln Val Tyr Gln Lys Phe Glu Lys Met Leu Ile Asp 980 985 990 Lys Leu Asn Phe Leu Val Phe Lys Glu Asn Lys Pro Thr Glu Pro Gly 995 1000 1005 Gly Val Leu Lys Ala Tyr Gln Leu Thr Asp Glu Phe Gln Ser Phe 1010 1015 1020 Glu Lys Leu Ser Lys Gln Thr Gly Phe Leu Phe Tyr Val Pro Ser 1025 1030 1035 Trp Asn Thr Ser Lys Ile Asp Pro Arg Thr Gly Phe Ile Asp Phe 1040 1045 1050 Leu His Pro Ala Tyr Glu Asn Ile Glu Lys Ala Lys Gln Trp Ile 1055 1060 1065 Asn Lys Phe Asp Ser Ile Arg Phe Asn Ser Lys Met Asp Trp Phe 1070 1075 1080 Glu Phe Thr Ala Asp Thr Arg Lys Phe Ser Glu Asn Leu Met Leu 1085 1090 1095 Gly Lys Asn Arg Val Trp Val Ile Cys Thr Thr Asn Val Glu Arg 1100 1105 1110 Tyr Phe Thr Ser Lys Thr Ala Asn Ser Ser Ile Gln Tyr Asn Ser 1115 1120 1125 Ile Gln Ile Thr Glu Lys Leu Lys Glu Leu Phe Val Asp Ile Pro 1130 1135 1140 Phe Ser Asn Gly Gln Asp Leu Lys Pro Glu Ile Leu Arg Lys Asn 1145 1150 1155 Asp Ala Val Phe Phe Lys Ser Leu Leu Phe Tyr Ile Lys Thr Thr 1160 1165 1170 Leu Ser Leu Arg Gln Asn Asn Gly Lys Lys Gly Glu Glu Glu Lys 1175 1180 1185 Asp Phe Ile Leu Ser Pro Val Val Asp Ser Lys Gly Arg Phe Phe 1190 1195 1200 Asn Ser Leu Glu Ala Ser Asp Asp Glu Pro Lys Asp Ala Asp Ala 1205 1210 1215 Asn Gly Ala Tyr His Ile Ala Leu Lys Gly Leu Met Asn Leu Leu 1220 1225 1230 Val Leu Asn Glu Thr Lys Glu Glu Asn Leu Ser Arg Pro Lys Trp 1235 1240 1245 Lys Ile Lys Asn Lys Asp Trp Leu Glu Phe Val Trp Glu Arg Asn 1250 1255 1260 Arg <210> 39 <211> 1373 <212> PRT <213> Moraxella bovoculi <400> 39 Met Leu Phe Gln Asp Phe Thr His Leu Tyr Pro Leu Ser Lys Thr Val 1 5 10 15 Arg Phe Glu Leu Phe Ile Asp Arg Thr Leu Glu His Ile His Ala Lys 20 25 30 Asn Phe Leu Ser Gln Asp Glu Thr Met Ala Asp Met His Gln Lys Val 35 40 45 Lys Val Ile Leu Asp Asp Tyr His Arg Asp Phe Ile Ala Asp Met Met 50 55 60 Gly Glu Val Lys Leu Thr Lys Leu Ala Glu Phe Tyr Asp Val Tyr Leu 65 70 75 80 Lys Phe Arg Lys Asn Pro Lys Asp Asp Glu Leu Gln Lys Ala Gln Leu 85 90 95 Lys Asp Leu Gln Ala Val Leu Arg Lys Glu Ile Val Lys Pro Ile Gly 100 105 110 Asn Gly Gly Lys Tyr Lys Ala Gly Tyr Asp Arg Leu Phe Gly Ala Lys 115 120 125 Leu Phe Lys Asp Gly Lys Glu Leu Gly Asp Leu Ala Lys Phe Val Ile 130 135 140 Ala Gln Glu Gly Glu Ser Ser Pro Lys Leu Ala His Leu Ala His Phe 145 150 155 160 Glu Lys Phe Ser Thr Tyr Phe Thr Gly Phe His Asp Asn Arg Lys Asn 165 170 175 Met Tyr Ser Asp Glu Asp Lys His Thr Ala Ile Ala Tyr Arg Leu Ile 180 185 190 His Glu Asn Leu Pro Arg Phe Ile Asp Asn Leu Gln Ile Leu Thr Thr 195 200 205 Ile Lys Gln Lys His Ser Ala Leu Tyr Asp Gln Ile Ile Asn Glu Leu 210 215 220 Thr Ala Ser Gly Leu Asp Val Ser Leu Ala Ser His Leu Asp Gly Tyr 225 230 235 240 His Lys Leu Leu Thr Gln Glu Gly Ile Thr Ala Tyr Asn Thr Leu Leu 245 250 255 Gly Gly Ile Ser Gly Glu Ala Gly Ser Pro Lys Ile Gln Gly Ile Asn 260 265 270 Glu Leu Ile Asn Ser His His Asn Gln His Cys His Lys Ser Glu Arg 275 280 285 Ile Ala Lys Leu Arg Pro Leu His Lys Gln Ile Leu Ser Asp Gly Met 290 295 300 Ser Val Ser Phe Leu Pro Ser Lys Phe Ala Asp Asp Ser Glu Met Cys 305 310 315 320 Gln Ala Val Asn Glu Phe Tyr Arg His Tyr Ala Asp Val Phe Ala Lys 325 330 335 Val Gln Ser Leu Phe Asp Gly Phe Asp Asp His Gln Lys Asp Gly Ile 340 345 350 Tyr Val Glu His Lys Asn Leu Asn Glu Leu Ser Lys Gln Ala Phe Gly 355 360 365 Asp Phe Ala Leu Leu Gly Arg Val Leu Asp Gly Tyr Tyr Val Asp Val 370 375 380 Val Asn Pro Glu Phe Asn Glu Arg Phe Ala Lys Ala Lys Thr Asp Asn 385 390 395 400 Ala Lys Ala Lys Leu Thr Lys Glu Lys Asp Lys Phe Ile Lys Gly Val 405 410 415 His Ser Leu Ala Ser Leu Glu Gln Ala Ile Glu His Tyr Thr Ala Arg 420 425 430 His Asp Asp Glu Ser Val Gln Ala Gly Lys Leu Gly Gln Tyr Phe Lys 435 440 445 His Gly Leu Ala Gly Val Asp Asn Pro Ile Gln Lys Ile His Asn Asn 450 455 460 His Ser Thr Ile Lys Gly Phe Leu Glu Arg Glu Arg Pro Ala Gly Glu 465 470 475 480 Arg Ala Leu Pro Lys Ile Lys Ser Gly Lys Asn Pro Glu Met Thr Gln 485 490 495 Leu Arg Gln Leu Lys Glu Leu Leu Asp Asn Ala Leu Asn Val Ala His 500 505 510 Phe Ala Lys Leu Leu Thr Thr Lys Thr Thr Leu Asp Asn Gln Asp Gly 515 520 525 Asn Phe Tyr Gly Glu Phe Gly Val Leu Tyr Asp Glu Leu Ala Lys Ile 530 535 540 Pro Thr Leu Tyr Asn Lys Val Arg Asp Tyr Leu Ser Gln Lys Pro Phe 545 550 555 560 Ser Thr Glu Lys Tyr Lys Leu Asn Phe Gly Asn Pro Thr Leu Leu Asn 565 570 575 Gly Trp Asp Leu Asn Lys Glu Lys Asp Asn Phe Gly Val Ile Leu Gln 580 585 590 Lys Asp Gly Cys Tyr Tyr Leu Ala Leu Leu Asp Lys Ala His Lys Lys 595 600 605 Val Phe Asp Asn Ala Pro Asn Thr Gly Lys Ser Ile Tyr Gln Lys Met 610 615 620 Ile Tyr Lys Tyr Leu Glu Val Arg Lys Gln Phe Pro Lys Val Phe Phe 625 630 635 640 Ser Lys Glu Ala Ile Ala Ile Asn Tyr His Pro Ser Lys Glu Leu Val 645 650 655 Glu Ile Lys Asp Lys Gly Arg Gln Arg Ser Asp Asp Glu Arg Leu Lys 660 665 670 Leu Tyr Arg Phe Ile Leu Glu Cys Leu Lys Ile His Pro Lys Tyr Asp 675 680 685 Lys Lys Phe Glu Gly Ala Ile Gly Asp Ile Gln Leu Phe Lys Lys Asp 690 695 700 Lys Lys Gly Arg Glu Val Pro Ile Ser Glu Lys Asp Leu Phe Lys Asp 705 710 715 720 Ile Asn Gly Ile Phe Ser Ser Lys Pro Lys Leu Glu Met Glu Asp Phe 725 730 735 Phe Ile Gly Glu Phe Lys Arg Tyr Asn Pro Ser Gln Asp Leu Val Asp 740 745 750 Gln Tyr Asn Ile Tyr Lys Lys Ile Asp Ser Asn Asp Asn Arg Lys Lys 755 760 765 Glu Asn Phe Tyr Asn Asn His Pro Lys Phe Lys Lys Asp Leu Val Arg 770 775 780 Tyr Tyr Tyr Glu Ser Met Cys Lys His Glu Glu Trp Glu Glu Ser Phe 785 790 795 800 Glu Phe Ser Lys Lys Leu Gln Asp Ile Gly Cys Tyr Val Asp Val Asn 805 810 815 Glu Leu Phe Thr Glu Ile Glu Thr Arg Arg Leu Asn Tyr Lys Ile Ser 820 825 830 Phe Cys Asn Ile Asn Ala Asp Tyr Ile Asp Glu Leu Val Glu Gln Gly 835 840 845 Gln Leu Tyr Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ser Pro Lys Ala 850 855 860 His Gly Lys Pro Asn Leu His Thr Leu Tyr Phe Lys Ala Leu Phe Ser 865 870 875 880 Glu Asp Asn Leu Ala Asp Pro Ile Tyr Lys Leu Asn Gly Glu Ala Gln 885 890 895 Ile Phe Tyr Arg Lys Ala Ser Leu Asp Met Asn Glu Thr Thr Ile His 900 905 910 Arg Ala Gly Glu Val Leu Glu Asn Lys Asn Pro Asp Asn Pro Lys Lys 915 920 925 Arg Gln Phe Val Tyr Asp Ile Ile Lys Asp Lys Arg Tyr Thr Gln Lys 930 935 940 Asp Phe Met Leu His Val Pro Ile Thr Met Asn Phe Gly Val Gln Gly 945 950 955 960 Met Thr Ile Lys Glu Phe Asn Lys Lys Val Asn Gln Ser Ile Gln Gln 965 970 975 Tyr Asp Glu Val Asn Val Ile Gly Ile Asp Arg Gly Glu Arg His Leu 980 985 990 Leu Tyr Leu Thr Val Ile Asn Ser Lys Gly Glu Ile Leu Glu Gln Cys 995 1000 1005 Ser Leu Asn Asp Ile Thr Thr Ala Ser Ala Asn Gly Thr Gln Met 1010 1015 1020 Thr Thr Pro Tyr His Lys Ile Leu Asp Lys Arg Glu Ile Glu Arg 1025 1030 1035 Leu Asn Ala Arg Val Gly Trp Gly Glu Ile Glu Thr Ile Lys Glu 1040 1045 1050 Leu Lys Ser Gly Tyr Leu Ser His Val Val His Gln Ile Ser Gln 1055 1060 1065 Leu Met Leu Lys Tyr Asn Ala Ile Val Val Leu Glu Asp Leu Asn 1070 1075 1080 Phe Gly Phe Lys Arg Gly Arg Phe Lys Val Glu Lys Gln Ile Tyr 1085 1090 1095 Gln Asn Phe Glu Asn Ala Leu Ile Lys Lys Leu Asn His Leu Val 1100 1105 1110 Leu Lys Asp Lys Ala Asp Asp Glu Ile Gly Ser Tyr Lys Asn Ala 1115 1120 1125 Leu Gln Leu Thr Asn Asn Phe Thr Asp Leu Lys Ser Ile Gly Lys 1130 1135 1140 Gln Thr Gly Phe Leu Phe Tyr Val Pro Ala Trp Asn Thr Ser Lys 1145 1150 1155 Ile Asp Pro Glu Thr Gly Phe Val Asp Leu Leu Lys Pro Arg Tyr 1160 1165 1170 Glu Asn Ile Gln Ala Ser Gln Ala Phe Phe Gly Lys Phe Asp Lys 1175 1180 1185 Ile Cys Tyr Asn Ala Asp Lys Asp Tyr Phe Glu Phe His Ile Asp 1190 1195 1200 Tyr Ala Lys Phe Thr Asp Lys Ala Lys Asn Ser Arg Gln Ile Trp 1205 1210 1215 Thr Ile Cys Ser His Gly Asp Lys Arg Tyr Val Tyr Asp Lys Thr 1220 1225 1230 Ala Asn Gln Asn Lys Gly Ala Ala Lys Gly Ile Asn Val Asn Asp 1235 1240 1245 Ile Leu Lys Ser Leu Phe Ala Arg His His Ile Asn Glu Lys Gln 1250 1255 1260 Pro Asn Leu Val Met Asp Ile Cys Gln Asn Asn Asp Lys Glu Phe 1265 1270 1275 His Lys Ser Leu Met Tyr Leu Leu Lys Thr Leu Leu Ala Leu Arg 1280 1285 1290 Tyr Ser Asn Ala Ser Ser Asp Glu Asp Phe Ile Leu Ser Pro Val 1295 1300 1305 Ala Asn Asp Glu Gly Val Phe Phe Asn Ser Ala Leu Ala Asp Asp 1310 1315 1320 Thr Gln Pro Gln Asn Ala Asp Ala Asn Gly Ala Tyr His Ile Ala 1325 1330 1335 Leu Lys Gly Leu Trp Leu Leu Asn Glu Leu Lys Asn Ser Asp Asp 1340 1345 1350 Leu Asn Lys Val Lys Leu Ala Ile Asp Asn Gln Thr Trp Leu Asn 1355 1360 1365 Phe Ala Gln Asn Arg 1370 <210> 40 <211> 1352 <212> PRT <213> Parcubacteria bacteria <400> 40 Met Glu Asn Ile Phe Asp Gln Phe Ile Gly Lys Tyr Ser Leu Ser Lys 1 5 10 15 Thr Leu Arg Phe Glu Leu Lys Pro Val Gly Lys Thr Glu Asp Phe Leu 20 25 30 Lys Ile Asn Lys Val Phe Glu Lys Asp Gln Thr Ile Asp Asp Ser Tyr 35 40 45 Asn Gln Ala Lys Phe Tyr Phe Asp Ser Leu His Gln Lys Phe Ile Asp 50 55 60 Ala Ala Leu Ala Ser Asp Lys Thr Ser Glu Leu Ser Phe Gln Asn Phe 65 70 75 80 Ala Asp Val Leu Glu Lys Gln Asn Lys Ile Ile Leu Asp Lys Lys Arg 85 90 95 Glu Met Gly Ala Leu Arg Lys Arg Asp Lys Asn Ala Val Gly Ile Asp 100 105 110 Arg Leu Gln Lys Glu Ile Asn Asp Ala Glu Asp Ile Ile Gln Lys Glu 115 120 125 Lys Glu Lys Ile Tyr Lys Asp Val Arg Thr Leu Phe Asp Asn Glu Ala 130 135 140 Glu Ser Trp Lys Thr Tyr Tyr Gln Glu Arg Glu Val Asp Gly Lys Lys 145 150 155 160 Ile Thr Glu Ser Lys Ala Asp Leu Lys Gln Lys Gly Ala Asp Phe Leu 165 170 175 Thr Ala Ala Gly Ile Leu Lys Val Leu Lys Tyr Glu Phe Pro Glu Glu 180 185 190 Lys Glu Lys Glu Phe Gln Ala Lys Asn Gln Pro Ser Leu Phe Val Glu 195 200 205 Glu Lys Glu Asn Pro Gly Gln Lys Arg Tyr Ile Phe Asp Ser Phe Asp 210 215 220 Lys Phe Ala Gly Tyr Leu Thr Lys Phe Gln Gln Thr Lys Lys Asn Leu 225 230 235 240 Tyr Ala Ala Asp Gly Thr Ser Thr Ala Val Ala Thr Arg Ile Ala Asp 245 250 255 Asn Phe Ile Ile Phe His Gln Asn Thr Lys Val Phe Arg Asp Lys Tyr 260 265 270 Lys Asn Asn His Thr Asp Leu Gly Phe Asp Glu Glu Asn Ile Phe Glu 275 280 285 Ile Glu Arg Tyr Lys Asn Cys Leu Leu Gln Arg Glu Ile Glu His Ile 290 295 300 Lys Asn Glu Asn Ser Tyr Asn Lys Ile Ile Gly Arg Ile Asn Lys Lys 305 310 315 320 Ile Lys Glu Tyr Arg Asp Gln Lys Ala Lys Asp Thr Lys Leu Thr Lys 325 330 335 Ser Asp Phe Pro Phe Phe Lys Asn Leu Asp Lys Gln Ile Leu Gly Glu 340 345 350 Val Glu Lys Glu Lys Gln Leu Ile Glu Lys Thr Arg Glu Lys Thr Glu 355 360 365 Glu Asp Val Leu Ile Glu Arg Phe Lys Glu Phe Ile Glu Asn Asn Glu 370 375 380 Glu Arg Phe Thr Ala Ala Lys Lys Leu Met Asn Ala Phe Cys Asn Gly 385 390 395 400 Glu Phe Glu Ser Glu Tyr Glu Gly Ile Tyr Leu Lys Asn Lys Ala Ile 405 410 415 Asn Thr Ile Ser Arg Arg Trp Phe Val Ser Asp Arg Asp Phe Glu Leu 420 425 430 Lys Leu Pro Gln Gln Lys Ser Lys Asn Lys Ser Glu Lys Asn Glu Pro 435 440 445 Lys Val Lys Lys Phe Ile Ser Ile Ala Glu Ile Lys Asn Ala Val Glu 450 455 460 Glu Leu Asp Gly Asp Ile Phe Lys Ala Val Phe Tyr Asp Lys Lys Ile 465 470 475 480 Ile Ala Gln Gly Gly Ser Lys Leu Glu Gln Phe Leu Val Ile Trp Lys 485 490 495 Tyr Glu Phe Glu Tyr Leu Phe Arg Asp Ile Glu Arg Glu Asn Gly Glu 500 505 510 Lys Leu Leu Gly Tyr Asp Ser Cys Leu Lys Ile Ala Lys Gln Leu Gly 515 520 525 Ile Phe Pro Gln Glu Lys Glu Ala Arg Glu Lys Ala Thr Ala Val Ile 530 535 540 Lys Asn Tyr Ala Asp Ala Gly Leu Gly Ile Phe Gln Met Met Lys Tyr 545 550 555 560 Phe Ser Leu Asp Asp Lys Asp Arg Lys Asn Thr Pro Gly Gln Leu Ser 565 570 575 Thr Asn Phe Tyr Ala Glu Tyr Asp Gly Tyr Tyr Lys Asp Phe Glu Phe 580 585 590 Ile Lys Tyr Tyr Asn Glu Phe Arg Asn Phe Ile Thr Lys Lys Pro Phe 595 600 605 Asp Glu Asp Lys Ile Lys Leu Asn Phe Glu Asn Gly Ala Leu Leu Lys 610 615 620 Gly Trp Asp Glu Asn Lys Glu Tyr Asp Phe Met Gly Val Ile Leu Lys 625 630 635 640 Lys Glu Gly Arg Leu Tyr Leu Gly Ile Met His Lys Asn His Arg Lys 645 650 655 Leu Phe Gln Ser Met Gly Asn Ala Lys Gly Asp Asn Ala Asn Arg Tyr 660 665 670 Gln Lys Met Ile Tyr Lys Gln Ile Ala Asp Ala Ser Lys Asp Val Pro 675 680 685 Arg Leu Leu Leu Thr Ser Lys Lys Ala Met Glu Lys Phe Lys Pro Ser 690 695 700 Gln Glu Ile Leu Arg Ile Lys Lys Glu Lys Thr Phe Lys Arg Glu Ser 705 710 715 720 Lys Asn Phe Ser Leu Arg Asp Leu His Ala Leu Ile Glu Tyr Tyr Arg 725 730 735 Asn Cys Ile Pro Gln Tyr Ser Asn Trp Ser Phe Tyr Asp Phe Gln Phe 740 745 750 Gln Asp Thr Gly Lys Tyr Gln Asn Ile Lys Glu Phe Thr Asp Asp Val 755 760 765 Gln Lys Tyr Gly Tyr Lys Ile Ser Phe Arg Asp Ile Asp Asp Glu Tyr 770 775 780 Ile Asn Gln Ala Leu Asn Glu Gly Lys Met Tyr Leu Phe Glu Val Val 785 790 795 800 Asn Lys Asp Ile Tyr Asn Thr Lys Asn Gly Ser Lys Asn Leu His Thr 805 810 815 Leu Tyr Phe Glu His Ile Leu Ser Ala Glu Asn Leu Asn Asp Pro Val 820 825 830 Phe Lys Leu Ser Gly Met Ala Glu Ile Phe Gln Arg Gln Pro Ser Val 835 840 845 Asn Glu Arg Glu Lys Ile Thr Thr Gln Lys Asn Gln Cys Ile Leu Asp 850 855 860 Lys Gly Asp Arg Ala Tyr Lys Tyr Arg Arg Tyr Thr Glu Lys Lys Ile 865 870 875 880 Methionine, Phenylalanine, Histidine, Methionine, Serine, Leucine, Valine, Leucine, Asparagine, Threonine, Glycine, Lysine, Glycine, Glutamic Acid, Isoleucine, Lysine 885 890 895 Glutamine, Valine, Glutamine, Phenylalanine, Asparagine, Lysine, Isoleucine, Isoleucine, Asparagine, Glutamine, Arginine, Isoleucine, Serine, Serine, Serine, Aspartic Acid 900 905 910 Asparagine, Glutamic Acid, Methionine, Arginine, Valine, Asparagine, Valine, Isoleucine, Glycine, Isoleucine, Aspartic Acid, Arginine, Glycine, Glutamic Acid, Lysine, Asparagine 915 920 925 Leucine, Leucine, Tyrosine, Tyrosine, Serine, Valine, Valine, Lysine, Glutamine, Asparagine, Glycine, Glutamic Acid, Isoleucine, Isoleucine, Glutamic Acid, Glutamine 930 935 940 Alanine, Serine, Leucine, Asparagine, Glutamic Acid, Isoleucine, Asparagine, Glycine, Valine, Asparagine, Tyrosine, Arginine, Aspartic Acid, Lysine, Leucine, Isoleucine 945 950 955 960 Glutamic Acid, Arginine, Glutamic Acid, Lysine, Glutamic Acid, Arginine, Leucine, Lysine, Asparagine, Arginine, Glutamine, Serine, Tryptophan, Lysine, Proline, Valine 965 970 975 Valine, Lysine, Isoleucine, Lysine, Aspartic Acid, Leucine, Lysine, Lysine, Glycine, Tyrosine, Isoleucine, Serine, Histidine, Valine, Isoleucine, Histidine 980 985 990 Lysine, Isoleucine, Cysteine, Glutamine, Leucine, Isoleucine, Glutamic Acid, Lysine, Tyrosine, Serine, Alanine, Isoleucine, Valine, Valine, Leucine, Glutamic Acid 995 1000 1005 Aspartic Acid, Leucine, Asparagine, Methionine, Arginine, Phenylalanine, Lysine, Glutamine, Isoleucine, Arginine, Glycine, Glycine, Isoleucine, Glutamic Acid, Arginine 1010 1015 1020 Serine, Valine, Tyrosine, Glutamine, Glutamine, Phenylalanine, Glutamic Acid, Lysine, Alanine, Leucine, Isoleucine, Aspartic Acid, Lysine, Leucine, Glycine 1025 1030 1035 Tyr Leu Val Phe Lys Asp Asn Arg Asp Leu Arg Ala Pro Gly Gly 1040 1045 1050 Val Leu Asn Gly Tyr Gln Leu Ser Ala Pro Phe Val Ser Phe Glu 1055 1060 1065 Lys Met Arg Lys Gln Thr Gly Ile Leu Phe Tyr Thr Gln Ala Glu 1070 1075 1080 Tyr Thr Ser Lys Thr Asp Pro Ile Thr Gly Phe Arg Lys Asn Val 1085 1090 1095 Tyr Ile Ser Asn Ser Ala Ser Leu Asp Lys Ile Lys Glu Ala Val 1100 1105 1110 Lys Lys Phe Asp Ala Ile Gly Trp Asp Gly Lys Glu Gln Ser Tyr 1115 1120 1125 Phe Phe Lys Tyr Asn Pro Tyr Asn Leu Ala Asp Glu Lys Tyr Lys 1130 1135 1140 Asn Ser Thr Val Ser Lys Glu Trp Ala Ile Phe Ala Ser Ala Pro 1145 1150 1155 Arg Ile Arg Arg Gln Lys Gly Glu Asp Gly Tyr Trp Lys Tyr Asp 1160 1165 1170 Arg Val Lys Val Asn Glu Glu Phe Glu Lys Leu Leu Lys Val Trp 1175 1180 1185 Asn Phe Val Asn Pro Lys Ala Thr Asp Ile Lys Gln Glu Ile Ile 1190 1195 1200 Lys Lys Ile Lys Ala Gly Asp Leu Gln Gly Glu Lys Glu Leu Asp 1205 1210 1215 Gly Arg Leu Arg Asn Phe Trp His Ser Phe Ile Tyr Leu Phe Asn 1220 1225 1230 Leu Val Leu Glu Leu Arg Asn Ser Phe Ser Leu Gln Ile Lys Ile 1235 1240 1245 Lys Ala Gly Glu Val Ile Ala Val Asp Glu Gly Val Asp Phe Ile 1250 1255 1260 Ala Ser Pro Val Lys Pro Phe Phe Thr Thr Pro Asn Pro Tyr Ile 1265 1270 1275 Pro Ser Asn Leu Cys Trp Leu Ala Val Glu Asn Ala Asp Ala Asn 1280 1285 1290 Gly Ala Tyr Asn Ile Ala Arg Lys Gly Val Met Ile Leu Lys Lys 1295 1300 1305 Ile Arg Glu His Ala Lys Lys Asp Pro Glu Phe Lys Lys Leu Pro 1310 1315 1320 Asn Leu Phe Ile Ser Asn Ala Glu Trp Asp Glu Ala Ala Arg Asp 1325 1330 1335 Trp Gly Lys Tyr Ala Gly Thr Thr Ala Leu Asn Leu Asp His 1340 1345 1350 <210> 41 <211> 1260 <212> PRT <213> Porphyromonas crevioricanis <400> 41 Met Asp Ser Leu Lys Asp Phe Thr Asn Leu Tyr Pro Val Ser Lys Thr 1 5 10 15 Leu Arg Phe Glu Leu Lys Pro Val Gly Lys Thr Leu Glu Asn Ile Glu 20 25 30 Lys Ala Gly Ile Leu Lys Glu Asp Glu His Arg Ala Glu Ser Tyr Arg 35 40 45 Arg Val Lys Lys Ile Ile Asp Thr Tyr His Lys Val Phe Ile Asp Ser 50 55 60 Ser Leu Glu Asn Met Ala Lys Met Gly Ile Glu Asn Glu Ile Lys Ala 65 70 75 80 Met Leu Gln Ser Phe Cys Glu Leu Tyr Lys Lys Asp His Arg Thr Glu 85 90 95 Gly Glu Asp Lys Ala Leu Asp Lys Ile Arg Ala Val Leu Arg Gly Leu 100 105 110 Ile Val Gly Ala Phe Thr Gly Val Cys Gly Arg Arg Glu Asn Thr Val 115 120 125 Gln Asn Glu Lys Tyr Glu Ser Leu Phe Lys Glu Lys Leu Ile Lys Glu 130 135 140 Ile Leu Pro Asp Phe Val Leu Ser Thr Glu Ala Glu Ser Leu Pro Phe 145 150 155 160 Ser Val Glu Glu Ala Thr Arg Ser Leu Lys Glu Phe Asp Ser Phe Thr 165 170 175 Ser Tyr Phe Ala Gly Phe Tyr Glu Asn Arg Lys Asn Ile Tyr Ser Thr 180 185 190 Lys Pro Gln Ser Thr Ala Ile Ala Tyr Arg Leu Ile His Glu Asn Leu 195 200 205 Pro Lys Phe Ile Asp Asn Ile Leu Val Phe Gln Lys Ile Lys Glu Pro 210 215 220 Ile Ala Lys Glu Leu Glu His Ile Arg Ala Asp Phe Ser Ala Gly Gly 225 230 235 240 Tyr Ile Lys Lys Asp Glu Arg Leu Glu Asp Ile Phe Ser Leu Asn Tyr 245 250 255 Tyr Ile His Val Leu Ser Gln Ala Gly Ile Glu Lys Tyr Asn Ala Leu 260 265 270 Ile Gly Lys Ile Val Thr Glu Gly Asp Gly Glu Met Lys Gly Leu Asn 275 280 285 Glu His Ile Asn Leu Tyr Asn Gln Gln Arg Gly Arg Glu Asp Arg Leu 290 295 300 Pro Leu Phe Arg Pro Leu Tyr Lys Gln Ile Leu Ser Asp Arg Glu Gln 305 310 315 320 Leu Ser Tyr Leu Pro Glu Ser Phe Glu Lys Asp Glu Glu Leu Leu Arg 325 330 335 Ala Leu Lys Glu Phe Tyr Asp His Ile Ala Glu Asp Ile Leu Gly Arg 340 345 350 Thr Gln Gln Leu Met Thr Ser Ile Ser Glu Tyr Asp Leu Ser Arg Ile 355 360 365 Tyr Val Arg Asn Asp Ser Gln Leu Thr Asp Ile Ser Lys Lys Met Leu 370 375 380 Gly Asp Trp Asn Ala Ile Tyr Met Ala Arg Glu Arg Ala Tyr Asp His 385 390 395 400 Glu Gln Ala Pro Lys Arg Ile Thr Ala Lys Tyr Glu Arg Asp Arg Ile 405 410 415 Lys Ala Leu Lys Gly Glu Glu Ser Ile Ser Leu Ala Asn Leu Asn Ser 420 425 430 Cys Ile Ala Phe Leu Asp Asn Val Arg Asp Cys Arg Val Asp Thr Tyr 435 440 445 Leu Ser Thr Leu Gly Gln Lys Glu Gly Pro His Gly Leu Ser Asn Leu 450 455 460 Val Glu Asn Val Phe Ala Ser Tyr His Glu Ala Glu Gln Leu Leu Ser 465 470 475 480 Phe Pro Tyr Pro Glu Glu Asn Asn Leu Ile Gln Asp Lys Asp Asn Val 485 490 495 Val Leu Ile Lys Asn Leu Leu Asp Asn Ile Ser Asp Leu Gln Arg Phe 500 505 510 Leu Lys Pro Leu Trp Gly Met Gly Asp Glu Pro Asp Lys Asp Glu Arg 515 520 525 Phe Tyr Gly Glu Tyr Asn Tyr Ile Arg Gly Ala Leu Asp Gln Val Ile 530 535 540 Pro Leu Tyr Asn Lys Val Arg Asn Tyr Leu Thr Arg Lys Pro Tyr Ser 545 550 555 560 Thr Arg Lys Val Lys Leu Asn Phe Gly Asn Ser Gln Leu Leu Ser Gly 565 570 575 Trp Asp Arg Asn Lys Glu Lys Asp Asn Ser Cys Val Ile Leu Arg Lys 580 585 590 Gly Gln Asn Phe Tyr Leu Ala Ile Met Asn Asn Arg His Lys Arg Ser 595 600 605 Phe Glu Asn Lys Met Leu Pro Glu Tyr Lys Glu Gly Glu Pro Tyr Phe 610 615 620 Glu Lys Met Asp Tyr Lys Phe Leu Pro Asp Pro Asn Lys Met Leu Pro 625 630 635 640 Lys Val Phe Leu Ser Lys Lys Gly Ile Glu Ile Tyr Lys Pro Ser Pro 645 650 655 Lys Leu Leu Glu Gln Tyr Gly His Gly Thr His Lys Lys Gly Asp Thr 660 665 670 Phe Ser Met Asp Asp Leu His Glu Leu Ile Asp Phe Phe Lys His Ser 675 680 685 Ile Glu Ala His Glu Asp Trp Lys Gln Phe Gly Phe Lys Phe Ser Asp 690 695 700 Thr Ala Thr Tyr Glu Asn Val Ser Ser Phe Tyr Arg Glu Val Glu Asp 705 710 715 720 Gln Gly Tyr Lys Leu Ser Phe Arg Lys Val Ser Glu Ser Tyr Val Tyr 725 730 735 Ser Leu Ile Asp Gln Gly Lys Leu Tyr Leu Phe Gln Ile Tyr Asn Lys 740 745 750 Asp Phe Ser Pro Cys Ser Lys Gly Thr Pro Asn Leu His Thr Leu Tyr 755 760 765 Trp Arg Met Leu Phe Asp Glu Arg Asn Leu Ala Asp Val Ile Tyr Lys 770 775 780 Leu Asp Gly Lys Ala Glu Ile Phe Phe Arg Glu Lys Ser Leu Lys Asn 785 790 795 800 Asp His Pro Thr His Pro Ala Gly Lys Pro Ile Lys Lys Lys Ser Arg 805 810 815 Gln Lys Lys Gly Glu Glu Ser Leu Phe Glu Tyr Asp Leu Val Lys Asp 820 825 830 Arg Arg Tyr Thr Met Asp Lys Phe Gln Phe His Val Pro Ile Thr Met 835 840 845 Asn Phe Lys Cys Ser Ala Gly Ser Lys Val Asn Asp Met Val Asn Ala 850 855 860 His Ile Arg Glu Ala Lys Asp Met His Val Ile Gly Ile Asp Arg Gly 865 870 875 880 Glu Arg Asn Leu Leu Tyr Ile Cys Val Ile Asp Ser Arg Gly Thr Ile 885 890 895 Leu Asp Gln Ile Ser Leu Asn Thr Ile Asn Asp Ile Asp Tyr His Asp 900 905 910 Leu Leu Glu Ser Arg Asp Lys Asp Arg Gln Gln Glu His Arg Asn Trp 915 920 925 Gln Thr Ile Glu Gly Ile Lys Glu Leu Lys Gln Gly Tyr Leu Ser Gln 930 935 940 Ala Val His Arg Ile Ala Glu Leu Met Val Ala Tyr Lys Ala Val Val 945 950 955 960 Ala Leu Glu Asp Leu Asn Met Gly Phe Lys Arg Gly Arg Gln Lys Val 965 970 975 Glu Ser Ser Val Tyr Gln Gln Phe Glu Lys Gln Leu Ile Asp Lys Leu 980 985 990 Asn Tyr Leu Val Asp Lys Lys Lys Arg Pro Glu Asp Ile Gly Gly Leu 995 1000 1005 Leu Arg Ala Tyr Gln Phe Thr Ala Pro Phe Lys Ser Phe Lys Glu 1010 1015 1020 Met Gly Lys Gln Asn Gly Phe Leu Phe Tyr Ile Pro Ala Trp Asn 1025 1030 1035 Thr Ser Asn Ile Asp Pro Thr Thr Gly Phe Val Asn Leu Phe His 1040 1045 1050 Val Gln Tyr Glu Asn Val Asp Lys Ala Lys Ser Phe Phe Gln Lys 1055 1060 1065 Phe Asp Ser Ile Ser Tyr Asn Pro Lys Lys Asp Trp Phe Glu Phe 1070 1075 1080 Ala Phe Asp Tyr Lys Asn Phe Thr Lys Lys Ala Glu Gly Ser Arg 1085 1090 1095 Ser Met Trp Ile Leu Cys Thr His Gly Ser Arg Ile Lys Asn Phe 1100 1105 1110 Arg Asn Ser Gln Lys Asn Gly Gln Trp Asp Ser Glu Glu Phe Ala 1115 1120 1125 Leu Thr Glu Ala Phe Lys Ser Leu Phe Val Arg Tyr Glu Ile Asp 1130 1135 1140 Tyr Thr Ala Asp Leu Lys Thr Ala Ile Val Asp Glu Lys Gln Lys 1145 1150 1155 Asp Phe Phe Val Asp Leu Leu Lys Leu Phe Lys Leu Thr Val Gln 1160 1165 1170 Met Arg Asn Ser Trp Lys Glu Lys Asp Leu Asp Tyr Leu Ile Ser 1175 1180 1185 Pro Val Ala Gly Ala Asp Gly Arg Phe Phe Asp Thr Arg Glu Gly 1190 1195 1200 Asn Lys Ser Leu Pro Lys Asp Ala Asp Ala Asn Gly Ala Tyr Asn 1205 1210 1215 Ile Ala Leu Lys Gly Leu Trp Ala Leu Arg Gln Ile Arg Gln Thr 1220 1225 1230 Ser Glu Gly Gly Lys Leu Lys Leu Ala Ile Ser Asn Lys Glu Trp 1235 1240 1245 Leu Gln Phe Val Gln Glu Arg Ser Tyr Glu Lys Asp 1250 1255 1260 <210> 42 <211> 1324 <212> PRT <213> Prevotella disiens <400> 42 Met Glu Asn Tyr Gln Glu Phe Thr Asn Leu Phe Gln Leu Asn Lys Thr 1 5 10 15 Leu Arg Phe Glu Leu Lys Pro Ile Gly Lys Thr Cys Glu Leu Leu Glu 20 25 30 Glu Gly Lys Ile Phe Ala Ser Gly Ser Phe Leu Glu Lys Asp Lys Val 35 40 45 Arg Ala Asp Asn Val Ser Tyr Val Lys Lys Glu Ile Asp Lys Lys His 50 55 60 Lys Ile Phe Ile Glu Glu Thr Leu Ser Ser Phe Ser Ile Ser Asn Asp 65 70 75 80 Leu Leu Lys Gln Tyr Phe Asp Cys Tyr Asn Glu Leu Lys Ala Phe Lys 85 90 95 Lys Asp Cys Lys Ser Asp Glu Glu Glu Val Lys Lys Thr Ala Leu Arg 100 105 110 Asn Lys Cys Thr Ser Ile Gln Arg Ala Met Arg Glu Ala Ile Ser Gln 115 120 125 Ala Phe Leu Lys Ser Pro Gln Lys Lys Leu Leu Ala Ile Lys Asn Leu 130 135 140 Ile Glu Asn Val Phe Lys Ala Asp Glu Asn Val Gln His Phe Ser Glu 145 150 155 160 Phe Thr Ser Tyr Phe Ser Gly Phe Glu Thr Asn Arg Glu Asn Phe Tyr 165 170 175 Ser Asp Glu Glu Lys Ser Thr Ser Ile Ala Tyr Arg Leu Val His Asp 180 185 190 Asn Leu Pro Ile Phe Ile Lys Asn Ile Tyr Ile Phe Glu Lys Leu Lys 195 200 205 Glu Gln Phe Asp Ala Lys Thr Leu Ser Glu Ile Phe Glu Asn Tyr Lys 210 215 220 Leu Tyr Val Ala Gly Ser Ser Leu Asp Glu Val Phe Ser Leu Glu Tyr 225 230 235 240 Phe Asn Asn Thr Leu Thr Gln Lys Gly Ile Asp Asn Tyr Asn Ala Val 245 250 255 Ile Gly Lys Ile Val Lys Glu Asp Lys Gln Glu Ile Gln Gly Leu Asn 260 265 270 Glu His Ile Asn Leu Tyr Asn Gln Lys His Lys Asp Arg Arg Leu Pro 275 280 285 Phe Phe Ile Ser Leu Lys Lys Gln Ile Leu Ser Asp Arg Glu Ala Leu 290 295 300 Ser Trp Leu Pro Asp Met Phe Lys Asn Asp Ser Glu Val Ile Asp Ala 305 310 315 320 Leu Lys Gly Phe Tyr Ile Glu Asp Gly Phe Glu Asn Asn Val Leu Thr 325 330 335 Pro Leu Ala Thr Leu Leu Ser Ser Leu Asp Lys Tyr Asn Leu Asn Gly 340 345 350 Ile Phe Ile Arg Asn Asn Glu Ala Leu Ser Ser Leu Ser Gln Asn Val 355 360 365 Tyr Arg Asn Phe Ser Ile Asp Glu Ala Ile Asp Ala Gln Asn Ala Glu 370 375 380 Leu Gln Thr Phe Asn Asn Tyr Glu Leu Ile Ala Asn Ala Leu Arg Ala 385 390 395 400 Lys Ile Lys Lys Glu Thr Lys Gln Gly Arg Lys Ser Phe Glu Lys Tyr 405 410 415 Glu Glu Tyr Ile Asp Lys Lys Val Lys Ala Ile Asp Ser Leu Ser Ile 420 425 430 Gln Glu Ile Asn Glu Leu Val Glu Asn Tyr Val Ser Glu Phe Asn Ser 435 440 445 Asn Ser Gly Asn Met Pro Arg Lys Val Glu Asp Tyr Phe Ser Leu Met 450 455 460 Arg Lys Gly Asp Phe Gly Ser Asn Asp Leu Ile Glu Asn Ile Lys Thr 465 470 475 480 Lys Leu Ser Ala Ala Glu Lys Leu Leu Gly Thr Lys Tyr Gln Glu Thr 485 490 495 Ala Lys Asp Ile Phe Lys Lys Asp Glu Asn Ser Lys Leu Ile Lys Glu 500 505 510 Leu Leu Asp Ala Thr Lys Gln Phe Gln His Phe Ile Lys Pro Leu Leu 515 520 525 Gly Thr Gly Glu Glu Ala Asp Arg Asp Leu Val Phe Tyr Gly Asp Phe 530 535 540 Leu Pro Leu Tyr Glu Lys Phe Glu Glu Leu Thr Leu Leu Tyr Asn Lys 545 550 555 560 Val Arg Asn Arg Leu Thr Gln Lys Pro Tyr Ser Lys Asp Lys Ile Arg 565 570 575 Leu Cys Phe Asn Lys Pro Lys Leu Met Thr Gly Trp Val Asp Ser Lys 580 585 590 Thr Glu Lys Ser Asp Asn Gly Thr Gln Tyr Gly Gly Tyr Leu Phe Arg 595 600 605 Lys Lys Asn Glu Ile Gly Glu Tyr Asp Tyr Phe Leu Gly Ile Ser Ser 610 615 620 Lys Ala Gln Leu Phe Arg Lys Asn Glu Ala Val Ile Gly Asp Tyr Glu 625 630 635 640 Arg Leu Asp Tyr Tyr Gln Pro Lys Ala Asn Thr Ile Tyr Gly Ser Ala 645 650 655 Tyr Glu Gly Glu Asn Ser Tyr Lys Glu Asp Lys Lys Arg Leu Asn Lys 660 665 670 Val Ile Ile Ala Tyr Ile Glu Gln Ile Lys Gln Thr Asn Ile Lys Lys 675 680 685 Ser Ile Ile Glu Ser Ile Ser Lys Tyr Pro Asn Ile Ser Asp Asp Asp 690 695 700 Lys Val Thr Pro Ser Ser Leu Leu Glu Lys Ile Lys Lys Val Ser Ile 705 710 715 720 Asp Ser Tyr Asn Gly Ile Leu Ser Phe Lys Ser Phe Gln Ser Val Asn 725 730 735 Lys Glu Val Ile Asp Asn Leu Leu Lys Thr Ile Ser Pro Leu Lys Asn 740 745 750 Lys Ala Glu Phe Leu Asp Leu Ile Asn Lys Asp Tyr Gln Ile Phe Thr 755 760 765 Glu Val Gln Ala Val Ile Asp Glu Ile Cys Lys Gln Lys Thr Phe Ile 770 775 780 Tyr Phe Pro Ile Ser Asn Val Glu Leu Glu Lys Glu Met Gly Asp Lys 785 790 795 800 Asp Lys Pro Leu Cys Leu Phe Gln Ile Ser Asn Lys Asp Leu Ser Phe 805 810 815 Ala Lys Thr Phe Ser Ala Asn Leu Arg Lys Lys Arg Gly Ala Glu Asn 820 825 830 Leu His Thr Met Leu Phe Lys Ala Leu Met Glu Gly Asn Gln Asp Asn 835 840 845 Leu Asp Leu Gly Ser Gly Ala Ile Phe Tyr Arg Ala Lys Ser Leu Asp 850 855 860 Gly Asn Lys Pro Thr His Pro Ala Asn Glu Ala Ile Lys Cys Arg Asn 865 870 875 880 Val Ala Asn Lys Asp Lys Val Ser Leu Phe Thr Tyr Asp Ile Tyr Lys 885 890 895 Asn Arg Arg Tyr Met Glu Asn Lys Phe Leu Phe His Leu Ser Ile Val 900 905 910 Gln Asn Tyr Lys Ala Ala Asn Asp Ser Ala Gln Leu Asn Ser Ser Ala 915 920 925 Thr Glu Tyr Ile Arg Lys Ala Asp Asp Leu His Ile Ile Gly Ile Asp 930 935 940 Arg Gly Glu Arg Asn Leu Leu Tyr Tyr Ser Val Ile Asp Met Lys Gly 945 950 955 960 Asn Ile Val Glu Gln Asp Ser Leu Asn Ile Ile Arg Asn Asn Asp Leu 965 970 975 Glu Thr Asp Tyr His Asp Leu Leu Asp Lys Arg Glu Lys Glu Arg Lys 980 985 990 Ala Asn Arg Gln Asn Trp Glu Ala Val Glu Gly Ile Lys Asp Leu Lys 995 1000 1005 Lys Gly Tyr Leu Ser Gln Ala Val His Gln Ile Ala Gln Leu Met 1010 1015 1020 Leu Lys Tyr Asn Ala Ile Ile Ala Leu Glu Asp Leu Gly Gln Met 1025 1030 1035 Phe Val Thr Arg Gly Gln Lys Ile Glu Lys Ala Val Tyr Gln Gln 1040 1045 1050 Phe Glu Lys Ser Leu Val Asp Lys Leu Ser Tyr Leu Val Asp Lys 1055 1060 1065 Lys Arg Pro Tyr Asn Glu Leu Gly Gly Ile Leu Lys Ala Tyr Gln 1070 1075 1080 Leu Ala Ser Ser Ile Thr Lys Asn Asn Ser Asp Lys Gln Asn Gly 1085 1090 1095 Phe Leu Phe Tyr Val Pro Ala Trp Asn Thr Ser Lys Ile Asp Pro 1100 1105 1110 Val Thr Gly Phe Thr Asp Leu Leu Arg Pro Lys Ala Met Thr Ile 1115 1120 1125 Lys Glu Ala Gln Asp Phe Phe Gly Ala Phe Asp Asn Ile Ser Tyr 1130 1135 1140 Asn Asp Lys Gly Tyr Phe Glu Phe Glu Thr Asn Tyr Asp Lys Phe 1145 1150 1155 Lys Ile Arg Met Lys Ser Ala Gln Thr Arg Trp Thr Ile Cys Thr 1160 1165 1170 Phe Gly Asn Arg Ile Lys Arg Lys Lys Asp Lys Asn Tyr Trp Asn 1175 1180 1185 Tyr Glu Glu Val Glu Leu Thr Glu Glu Phe Lys Lys Leu Phe Lys 1190 1195 1200 Asp Ser Asn Ile Asp Tyr Glu Asn Cys Asn Leu Lys Glu Glu Ile 1205 1210 1215 Gln Asn Lys Asp Asn Arg Lys Phe Phe Asp Asp Leu Ile Lys Leu 1220 1225 1230 Leu Gln Leu Thr Leu Gln Met Arg Asn Ser Asp Asp Lys Gly Asn 1235 1240 1245 Asp Tyr Ile Ile Ser Pro Val Ala Asn Ala Glu Gly Gln Phe Phe 1250 1255 1260 Asp Ser Arg Asn Gly Asp Lys Lys Leu Pro Leu Asp Ala Asp Ala 1265 1270 1275 Asn Gly Ala Tyr Asn Ile Ala Arg Lys Gly Leu Trp Asn Ile Arg 1280 1285 1290 Gln Ile Lys Gln Thr Lys Asn Lys Asp Asp Leu Asn Leu Ser Ile 1295 1300 1305 Ser Ser Thr Glu Trp Leu Asp Phe Val Arg Glu Lys Pro Tyr Leu 1310 1315 1320 Lys <210> 43 <211> 1484 <212> PRT <213> Peregrinibacteria bacteria <220> <221> misc_feature <222> (1073)..(1073) <223> Xaa can be any naturally occurring amino acid <400> 43 Met Ser Asn Phe Phe Lys Asn Phe Thr Asn Leu Tyr Glu Leu Ser Lys 1 5 10 15 Thr Leu Arg Phe Glu Leu Lys Pro Val Gly Asp Thr Leu Thr Asn Met 20 25 30 Lys Asp His Leu Glu Tyr Asp Glu Lys Leu Gln Thr Phe Leu Lys Asp 35 40 45 Gln Asn Ile Asp Asp Ala Tyr Gln Ala Leu Lys Pro Gln Phe Asp Glu 50 55 60 Ile His Glu Glu Phe Ile Thr Asp Ser Leu Glu Ser Lys Lys Ala Lys 65 70 75 80 Glu Ile Asp Phe Ser Glu Tyr Leu Asp Leu Phe Gln Glu Lys Lys Glu 85 90 95 Leu Asn Asp Ser Glu Lys Lys Leu Arg Asn Lys Ile Gly Glu Thr Phe 100 105 110 Asn Lys Ala Gly Glu Lys Trp Lys Lys Glu Lys Tyr Pro Gln Tyr Glu 115 120 125 Trp Lys Lys Gly Ser Lys Ile Ala Asn Gly Ala Asp Ile Leu Ser Cys 130 135 140 Gln Asp Met Leu Gln Phe Ile Lys Tyr Lys Asn Pro Glu Asp Glu Lys 145 150 155 160 Ile Lys Asn Tyr Ile Asp Asp Thr Leu Lys Gly Phe Phe Thr Tyr Phe 165 170 175 Gly Gly Phe Asn Gln Asn Arg Ala Asn Tyr Tyr Glu Thr Lys Lys Glu 180 185 190 Ala Ser Thr Ala Val Ala Thr Arg Ile Val His Glu Asn Leu Pro Lys 195 200 205 Phe Cys Asp Asn Val Ile Gln Phe Lys His Ile Ile Lys Arg Lys Lys 210 215 220 Asp Gly Thr Val Glu Lys Thr Glu Arg Lys Thr Glu Tyr Leu Asn Ala 225 230 235 240 Tyr Gln Tyr Leu Lys Asn Asn Asn Lys Ile Thr Gln Ile Lys Asp Ala 245 250 255 Glu Thr Glu Lys Met Ile Glu Ser Thr Pro Ile Ala Glu Lys Ile Phe 260 265 270 Asp Val Tyr Tyr Phe Ser Ser Cys Leu Ser Gln Lys Gln Ile Glu Glu 275 280 285 Tyr Asn Arg Ile Ile Gly His Tyr Asn Leu Leu Ile Asn Leu Tyr Asn 290 295 300 Gln Ala Lys Arg Ser Glu Gly Lys His Leu Ser Ala Asn Glu Lys Lys 305 310 315 320 Tyr Lys Asp Leu Pro Lys Phe Lys Thr Leu Tyr Lys Gln Ile Gly Cys 325 330 335 Gly Lys Lys Lys Asp Leu Phe Tyr Thr Ile Lys Cys Asp Thr Glu Glu 340 345 350 Glu Ala Asn Lys Ser Arg Asn Glu Gly Lys Glu Ser His Ser Val Glu 355 360 365 Glu Ile Ile Asn Lys Ala Gln Glu Ala Ile Asn Lys Tyr Phe Lys Ser 370 375 380 Asn Asn Asp Cys Glu Asn Ile Asn Thr Val Pro Asp Phe Ile Asn Tyr 385 390 395 400 Ile Leu Thr Lys Glu Asn Tyr Glu Gly Val Tyr Trp Ser Lys Ala Ala 405 410 415 Met Asn Thr Ile Ser Asp Lys Tyr Phe Ala Asn Tyr His Asp Leu Gln 420 425 430 Asp Arg Leu Lys Glu Ala Lys Val Phe Gln Lys Ala Asp Lys Lys Ser 435 440 445 Glu Asp Asp Ile Lys Ile Pro Glu Ala Ile Glu Leu Ser Gly Leu Phe 450 455 460 Gly Val Leu Asp Ser Leu Ala Asp Trp Gln Thr Thr Leu Phe Lys Ser 465 470 475 480 Ser Ile Leu Ser Asn Glu Lys Leu Lys Ile Ile Thr Asp Ser Gln Thr 485 490 495 Pro Ser Glu Ala Leu Leu Lys Met Ile Phe Asn Asp Ile Glu Lys Asn 500 505 510 Met Glu Ser Phe Leu Lys Glu Thr Asn Asp Ile Ile Thr Leu Lys Lys 515 520 525 Tyr Lys Gly Asn Lys Glu Gly Thr Glu Lys Ile Lys Gln Trp Phe Asp 530 535 540 Tyr Thr Leu Ala Ile Asn Arg Met Leu Lys Tyr Phe Leu Val Lys Glu 545 550 555 560 Asn Lys Ile Lys Gly Asn Ser Leu Asp Thr Asn Ile Ser Glu Ala Leu 565 570 575 Lys Thr Leu Ile Tyr Ser Asp Asp Ala Glu Trp Phe Lys Trp Tyr Asp 580 585 590 Ala Leu Arg Asn Tyr Leu Thr Gln Lys Pro Gln Asp Glu Ala Lys Glu 595 600 605 Asn Lys Leu Lys Leu Asn Phe Asp Asn Pro Ser Leu Ala Gly Gly Trp 610 615 620 Asp Val Asn Lys Glu Cys Ser Asn Phe Cys Val Ile Leu Lys Asp Lys 625 630 635 640 Asn Glu Lys Lys Tyr Leu Ala Met Ile Lys Lys Gly Glu Asn Thr Leu 645 650 655 Phe Gln Lys Glu Trp Thr Glu Gly Arg Gly Lys Asn Leu Thr Lys Lys 660 665 670 Ser Asn Pro Leu Phe Glu Ile Asn Asn Cys Glu Ile Leu Ser Lys Met 675 680 685 Glu Tyr Asp Phe Trp Ala Asp Val Ser Lys Met Ile Pro Lys Cys Ser 690 695 700 Thr Gln Leu Lys Ala Val Val Asn His Phe Lys Gln Ser Asp Asn Glu 705 710 715 720 Phe Ile Phe Pro Ile Gly Tyr Lys Val Thr Ser Gly Glu Lys Phe Arg 725 730 735 Glu Glu Cys Lys Ile Ser Lys Gln Asp Phe Glu Leu Asn Asn Lys Val 740 745 750 Phe Asn Lys Asn Glu Leu Ser Val Thr Ala Met Arg Tyr Asp Leu Ser 755 760 765 Ser Thr Gln Glu Lys Gln Tyr Ile Lys Ala Phe Gln Lys Glu Tyr Trp 770 775 780 Glu Leu Leu Phe Lys Gln Glu Lys Arg Asp Thr Lys Leu Thr Asn Asn 785 790 795 800 Glu Ile Phe Asn Glu Trp Ile Asn Phe Cys Asn Lys Lys Tyr Ser Glu 805 810 815 Leu Leu Ser Trp Glu Arg Lys Tyr Lys Asp Ala Leu Thr Asn Trp Ile 820 825 830 Asn Phe Cys Lys Tyr Phe Leu Ser Lys Tyr Pro Lys Thr Thr Leu Phe 835 840 845 Asn Tyr Ser Phe Lys Glu Ser Glu Asn Tyr Asn Ser Leu Asp Glu Phe 850 855 860 Tyr Arg Asp Val Asp Ile Cys Ser Tyr Lys Leu Asn Ile Asn Thr Thr 865 870 875 880 Ile Asn Lys Ser Ile Leu Asp Arg Leu Val Glu Glu Gly Lys Leu Tyr 885 890 895 Leucine Phenylalanine Glutamic acid Isoleucine Lysine Asparagine Glutamine Aspartic acid Serine Asparagine Aspartic acid Glycine Lysine Serine Isoleucine Glycine 900 905 910 Histidine Lysine Asparagine Asparagine Leucine Histidine Threonine Isoleucine Tyrosine Tryptophan Asparagine Alanine Isoleucine Phenylalanine Glutamic acid Asparagine 915 920 925 Phenylalanine Aspartic acid Asparagine Arginine Proline Lysine Leucine Asparagine Glycine Glutamic acid Alanine Glutamic acid Isoleucine Phenylalanine Tyrosine Arginine 930 935 940 Lysine Alanine Isoleucine Serine Lysine Aspartic acid Lysine Leucine Glycine Isoleucine Valine Lysine Glycine Lysine Lysine Threonine 945 950 955 960 Lysine Asparagine Glycine Threonine Tryptophan Isoleucine Isoleucine Lysine Asparagine Tyrosine Arginine Phenylalanine Serine Lysine Glutamic acid Lysine 965 970 975 Phenylalanine Isoleucine Leucine Histidine Valine Proline Isoleucine Threonine Leucine Asparagine Phenylalanine Cysteine Serine Asparagine Asparagine Glutamic acid 980 985 990 Tyrosine Valine Asparagine Aspartic acid Isoleucine Valine Asparagine Threonine Lysine Phenylalanine Tyrosine Asparagine Phenylalanine Serine Asparagine Leucine 995 1000 1005 Histidine Phenylalanine Leucine Glycine Isoleucine Aspartic acid Arginine Glycine Glutamic acid Lysine Histidine Leucine Alanine Tyrosine Tyrosine 1010 1015 1020 Serine Leucine Valine Asparagine Lysine Asparagine Glycine Glutamic acid Isoleucine Valine Aspartic acid Glutamine Glycine Threonine Leucine 1025 1030 1035 Asparagine Leucine Proline Phenylalanine Threonine Aspartic acid Lysine Aspartic acid Glycine Asparagine Glutamine Arginine Serine Isoleucine Lysine 1040 1045 1050 Lys Glu Lys Tyr Phe Tyr Asn Lys Gln Glu Asp Lys Trp Glu Ala 1055 1060 1065 Lys Glu Val Asp Xaa Trp Asn Tyr Asn Asp Leu Leu Asp Ala Met 1070 1075 1080 Ala Ser Asn Arg Asp Met Ala Arg Lys Asn Trp Gln Arg Ile Gly 1085 1090 1095 Thr Ile Lys Glu Ala Lys Asn Gly Tyr Val Ser Leu Val Ile Arg 1100 1105 1110 Lys Ile Ala Asp Leu Ala Val Asn Asn Glu Arg Pro Ala Phe Ile 1115 1120 1125 Val Leu Glu Asp Leu Asn Thr Gly Phe Lys Arg Ser Arg Gln Lys 1130 1135 1140 Ile Asp Lys Ser Val Tyr Gln Lys Phe Glu Leu Ala Leu Ala Lys 1145 1150 1155 Lys Leu Asn Phe Leu Val Asp Lys Asn Ala Lys Arg Asp Glu Ile 1160 1165 1170 Gly Ser Pro Thr Lys Ala Leu Gln Leu Thr Pro Pro Val Asn Asn 1175 1180 1185 Tyr Gly Asp Ile Glu Asn Lys Lys Gln Ala Gly Ile Met Leu Tyr 1190 1195 1200 Thr Arg Ala Asn Tyr Thr Ser Gln Thr Asp Pro Ala Thr Gly Trp 1205 1210 1215 Arg Lys Thr Ile Tyr Leu Lys Ala Gly Pro Glu Glu Thr Thr Tyr 1220 1225 1230 Lys Lys Asp Gly Lys Ile Lys Asn Lys Ser Val Lys Asp Gln Ile 1235 1240 1245 Ile Glu Thr Phe Thr Asp Ile Gly Phe Asp Gly Lys Asp Tyr Tyr 1250 1255 1260 Phe Glu Tyr Asp Lys Gly Glu Phe Val Asp Glu Lys Thr Gly Glu 1265 1270 1275 Ile Lys Pro Lys Lys Trp Arg Leu Tyr Ser Gly Glu Asn Gly Lys 1280 1285 1290 Ser Leu Asp Arg Phe Arg Gly Glu Arg Glu Lys Asp Lys Tyr Glu 1295 1300 1305 Trp Lys Ile Asp Lys Ile Asp Ile Val Lys Ile Leu Asp Asp Leu 1310 1315 1320 Phe Val Asn Phe Asp Lys Asn Ile Ser Leu Leu Lys Gln Leu Lys 1325 1330 1335 Glu Gly Val Glu Leu Thr Arg Asn Asn Glu His Gly Thr Gly Glu 1340 1345 1350 Ser Leu Arg Phe Ala Ile Asn Leu Ile Gln Gln Ile Arg Asn Thr 1355 1360 1365 Gly Asn Asn Glu Arg Asp Asn Asp Phe Ile Leu Ser Pro Val Arg 1370 1375 1380 Asp Glu Asn Gly Lys His Phe Asp Ser Arg Glu Tyr Trp Asp Lys 1385 1390 1395 Glu Thr Lys Gly Glu Lys Ile Ser Met Pro Ser Ser Gly Asp Ala 1400 1405 1410 Asn Gly Ala Phe Asn Ile Ala Arg Lys Gly Ile Ile Met Asn Ala 1415 1420 1425 His Ile Leu Ala Asn Ser Asp Ser Lys Asp Leu Ser Leu Phe Val 1430 1435 1440 Ser Asp Glu Glu Trp Asp Leu His Leu Asn Asn Lys Thr Glu Trp 1445 1450 1455 Lys Lys Gln Leu Asn Ile Phe Ser Ser Arg Lys Ala Met Ala Lys 1460 1465 1470 Arg Lys Lys Lys Arg Pro Ala Ala Thr Lys Lys 1475 1480 <210> 44 <211> 1245 <212> PRT <213> Porphyromonas macacae <400> 44 Methionine, Lysine, Threonine, Glutamine, Histidine, Phenylalanine, Phenylalanine, Glutamic acid, Aspartic acid, Phenylalanine, Threonine, Serine, Leucine, Tyrosine, Serine, Leucine 1 5 10 15 Serine, Lysine, Threonine, Isoleucine, Arginine, Phenylalanine, Glutamic acid, Leucine, Lysine, Proline, Isoleucine, Glycine, Lysine, Threonine, Leucine, Glutamic acid 20 25 30 Asparagine, Isoleucine, Lysine, Lysine, Asparagine, Glycine, Leucine, Isoleucine, Arginine, Arginine, Aspartic acid, Glutamic acid, Glutamine, Arginine, Leucine, Aspartic acid 35 40 45 Aspartic acid, Tyrosine, Glutamic acid, Lysine, Leucine, Lysine, Lysine, Valine, Isoleucine, Aspartic acid, Glutamic acid, Tyrosine, Histidine, Glutamic acid, Aspartic acid, Phenylalanine 50 55 60 Isoleucine, Alanine, Asparagine, Isoleucine, Leucine, Serine, Serine, Phenylalanine, Serine, Phenylalanine, Serine, Glutamic acid, Glutamic acid, Isoleucine, Leucine, Glutamine 65 70 75 80 Serine, Tyrosine, Isoleucine, Glutamine, Asparagine, Leucine, Serine, Isoleucine, Serine, Glutamic acid, Alanine, Arginine, Alanine, Lysine, Isoleucine, Glutamic acid 85 90 95 Lysine, Threonine, Methionine, Arginine, Aspartic acid, Threonine, Leucine, Alanine, Lysine, Alanine, Phenylalanine, Serine, Glutamic acid, Aspartic acid, Glutamic acid, Arginine 100 105 110 Tyrosine, Lysine, Serine, Isoleucine, Phenylalanine, Lysine, Lysine, Glutamic acid, Leucine, Valine, Lysine, Lysine, Aspartic acid, Isoleucine, Proline, Valine 115 120 125 Tryptophan, Cysteine, Proline, Alanine, Tyrosine, Lysine, Serine, Leucine, Cysteine, Lysine, Lysine, Phenylalanine, Aspartic acid, Asparagine, Phenylalanine, Threonine 130 135 140 Threonine, Serine, Leucine, Valine, Proline, Phenylalanine, Histidine, Glutamic acid, Asparagine, Arginine, Lysine, Asparagine, Leucine, Tyrosine, Threonine, Serine 145 150 155 160 Asn Glu Ile Thr Ala Ser Ile Pro Tyr Arg Ile Val His Val Asn Leu 165 170 175 Pro Lys Phe Ile Gln Asn Ile Glu Ala Leu Cys Glu Leu Gln Lys Lys 180 185 190 Met Gly Ala Asp Leu Tyr Leu Glu Met Met Glu Asn Leu Arg Asn Val 195 200 205 Trp Pro Ser Phe Val Lys Thr Pro Asp Asp Leu Cys Asn Leu Lys Thr 210 215 220 Tyr Asn His Leu Met Val Gln Ser Ser Ile Ser Glu Tyr Asn Arg Phe 225 230 235 240 Val Gly Gly Tyr Ser Thr Glu Asp Gly Thr Lys His Gln Gly Ile Asn 245 250 255 Glu Trp Ile Asn Ile Tyr Arg Gln Arg Asn Lys Glu Met Arg Leu Pro 260 265 270 Gly Leu Val Phe Leu His Lys Gln Ile Leu Ala Lys Val Asp Ser Ser 275 280 285 Ser Phe Ile Ser Asp Thr Leu Glu Asn Asp Asp Gln Val Phe Cys Val 290 295 300 Leu Arg Gln Phe Arg Lys Leu Phe Trp Asn Thr Val Ser Ser Lys Glu 305 310 315 320 Asp Asp Ala Ala Ser Leu Lys Asp Leu Phe Cys Gly Leu Ser Gly Tyr 325 330 335 Asp Pro Glu Ala Ile Tyr Val Ser Asp Ala His Leu Ala Thr Ile Ser 340 345 350 Lys Asn Ile Phe Asp Arg Trp Asn Tyr Ile Ser Asp Ala Ile Arg Arg 355 360 365 Lys Thr Glu Val Leu Met Pro Arg Lys Lys Glu Ser Val Glu Arg Tyr 370 375 380 Ala Glu Lys Ile Ser Lys Gln Ile Lys Lys Arg Gln Ser Tyr Ser Leu 385 390 395 400 Ala Glu Leu Asp Asp Leu Leu Ala His Tyr Ser Glu Glu Ser Leu Pro 405 410 415 Ala Gly Phe Ser Leu Leu Ser Tyr Phe Thr Ser Leu Gly Gly Gln Lys 420 425 430 Tyr Leu Val Ser Asp Gly Glu Val Ile Leu Tyr Glu Glu Gly Ser Asn 435 440 445 Ile Trp Asp Glu Val Leu Ile Ala Phe Arg Asp Leu Gln Val Ile Leu 450 455 460 Asp Lys Asp Phe Thr Glu Lys Lys Leu Gly Lys Asp Glu Glu Ala Val 465 470 475 480 Ser Val Ile Lys Lys Ala Leu Asp Ser Ala Leu Arg Leu Arg Lys Phe 485 490 495 Phe Asp Leu Leu Ser Gly Thr Gly Ala Glu Ile Arg Arg Asp Ser Ser 500 505 510 Phe Tyr Ala Leu Tyr Thr Asp Arg Met Asp Lys Leu Lys Gly Leu Leu 515 520 525 Lys Met Tyr Asp Lys Val Arg Asn Tyr Leu Thr Lys Lys Pro Tyr Ser 530 535 540 Ile Glu Lys Phe Lys Leu His Phe Asp Asn Pro Ser Leu Leu Ser Gly 545 550 555 560 Trp Asp Lys Asn Lys Glu Leu Asn Asn Leu Ser Val Ile Phe Arg Gln 565 570 575 Asn Gly Tyr Tyr Tyr Leu Gly Ile Met Thr Pro Lys Gly Lys Asn Leu 580 585 590 Phe Lys Thr Leu Pro Lys Leu Gly Ala Glu Glu Met Phe Tyr Glu Lys 595 600 605 Met Glu Tyr Lys Gln Ile Ala Glu Pro Met Leu Met Leu Pro Lys Val 610 615 620 Phe Phe Pro Lys Lys Thr Lys Pro Ala Phe Ala Pro Asp Gln Ser Val 625 630 635 640 Val Asp Ile Tyr Asn Lys Lys Thr Phe Lys Thr Gly Gln Lys Gly Phe 645 650 655 Asn Lys Lys Asp Leu Tyr Arg Leu Ile Asp Phe Tyr Lys Glu Ala Leu 660 665 670 Thr Val His Glu Trp Lys Leu Phe Asn Phe Ser Phe Ser Pro Thr Glu 675 680 685 Gln Tyr Arg Asn Ile Gly Glu Phe Phe Asp Glu Val Arg Glu Gln Ala 690 695 700 Tyr Lys Val Ser Met Val Asn Val Pro Ala Ser Tyr Ile Asp Glu Ala 705 710 715 720 Val Glu Asn Gly Lys Leu Tyr Leu Phe Gln Ile Tyr Asn Lys Asp Phe 725 730 735 Ser Pro Tyr Ser Lys Gly Ile Pro Asn Leu His Thr Leu Tyr Trp Lys 740 745 750 Ala Leu Phe Ser Glu Gln Asn Gln Ser Arg Val Tyr Lys Leu Cys Gly 755 760 765 Gly Gly Glu Leu Phe Tyr Arg Lys Ala Ser Leu His Met Gln Asp Thr 770 775 780 Thr Val His Pro Lys Gly Ile Ser Ile His Lys Lys Asn Leu Asn Lys 785 790 795 800 Lys Gly Glu Thr Ser Leu Phe Asn Tyr Asp Leu Val Lys Asp Lys Arg 805 810 815 Phe Thr Glu Asp Lys Phe Phe Phe His Val Pro Ile Ser Ile Asn Tyr 820 825 830 Lys Asn Lys Lys Ile Thr Asn Val Asn Gln Met Val Arg Asp Tyr Ile 835 840 845 Ala Gln Asn Asp Asp Leu Gln His Gly Ile Asp Arg Gly Glu Arg Asn 850 855 860 Leu Leu Tyr Ile Ser Arg Ile Asp Thr Arg Gly Asn Leu Leu Glu Gln 865 870 875 880 Phe Ser Leu Asn Val Ile Glu Ser Asp Lys Gly Asp Leu Arg Thr Asp 885 890 895 Tyr Gln Lys Ile Leu Gly Asp Arg Glu Gln Glu Arg Leu Arg Arg Arg 900 905 910 Gln Glu Trp Lys Ser Ile Glu Ser Ile Lys Asp Leu Lys Asp Gly Tyr 915 920 925 Met Ser Gln Val Val His Lys Ile Cys Asn Met Val Val Glu His Lys 930 935 940 Ala Ile Val Val Leu Glu Asn Leu Asn Leu Ser Phe Met Lys Gly Arg 945 950 955 960 Lys Lys Val Glu Lys Ser Val Tyr Glu Lys Phe Glu Arg Met Leu Val 965 970 975 Asp Lys Leu Asn Tyr Leu Val Val Asp Lys Lys Asn Leu Ser Asn Glu 980 985 990 Pro Gly Gly Leu Tyr Ala Ala Tyr Gln Leu Thr Asn Pro Leu Phe Ser 995 1000 1005 Phe Glu Glu Leu His Arg Tyr Pro Gln Ser Gly Ile Leu Phe Phe 1010 1015 1020 Val Asp Pro Trp Asn Thr Ser Leu Thr Asp Pro Ser Thr Gly Phe 1025 1030 1035 Val Asn Leu Leu Gly Arg Ile Asn Tyr Thr Asn Val Gly Asp Ala 1040 1045 1050 Arg Lys Phe Phe Asp Arg Phe Asn Ala Ile Arg Tyr Asp Gly Lys 1055 1060 1065 Gly Asn Ile Leu Phe Asp Leu Asp Leu Ser Arg Phe Asp Val Arg 1070 1075 1080 Val Glu Thr Gln Arg Lys Leu Trp Thr Leu Thr Thr Phe Gly Ser 1085 1090 1095 Arg Ile Ala Lys Ser Lys Lys Ser Gly Lys Trp Met Val Glu Arg 1100 1105 1110 Ile Glu Asn Leu Ser Leu Cys Phe Leu Glu Leu Phe Glu Gln Phe 1115 1120 1125 Asn Ile Gly Tyr Arg Val Glu Lys Asp Leu Lys Lys Ala Ile Leu 1130 1135 1140 Ser Gln Asp Arg Lys Glu Phe Tyr Val Arg Leu Ile Tyr Leu Phe 1145 1150 1155 Asn Leu Met Met Gln Ile Arg Asn Ser Asp Gly Glu Glu Asp Tyr 1160 1165 1170 Ile Leu Ser Pro Ala Leu Asn Glu Lys Asn Leu Gln Phe Asp Ser 1175 1180 1185 Arg Leu Ile Glu Ala Lys Asp Leu Pro Val Asp Ala Asp Ala Asn 1190 1195 1200 Gly Ala Tyr Asn Val Ala Arg Lys Gly Leu Met Val Val Gln Arg 1205 1210 1215 Ile Lys Arg Gly Asp His Glu Ser Ile His Arg Ile Gly Arg Ala 1220 1225 1230 Gln Trp Leu Arg Tyr Val Gln Glu Gly Ile Val Glu 1235 1240 1245 <210> 45 <211> 1250 <212> PRT <213> Smithella sp. <400> 45 Met Gln Thr Leu Phe Glu Asn Phe Thr Asn Gln Tyr Pro Val Ser Lys 1 5 10 15 Thr Leu Arg Phe Glu Leu Ile Pro Gln Gly Lys Thr Lys Asp Phe Ile 20 25 30 Glu Gln Lys Gly Leu Leu Lys Lys Asp Glu Asp Arg Ala Glu Lys Tyr 35 40 45 Lys Lys Val Lys Asn Ile Ile Asp Glu Tyr His Lys Asp Phe Ile Glu 50 55 60 Lys Ser Leu Asn Gly Leu Lys Leu Asp Gly Leu Glu Lys Tyr Lys Thr 65 70 75 80 Leu Tyr Leu Lys Gln Glu Lys Asp Asp Lys Asp Lys Lys Ala Phe Asp 85 90 95 Lys Glu Lys Glu Asn Leu Arg Lys Gln Ile Ala Asn Ala Phe Arg Asn 100 105 110 Asn Glu Lys Phe Lys Thr Leu Phe Ala Lys Glu Leu Ile Lys Asn Asp 115 120 125 Leu Met Ser Phe Ala Cys Glu Glu Asp Lys Lys Asn Val Lys Glu Phe 130 135 140 Glu Ala Phe Thr Thr Tyr Phe Thr Gly Phe His Gln Asn Arg Ala Asn 145 150 155 160 Met Tyr Val Ala Asp Glu Lys Arg Thr Ala Ile Ala Ser Arg Leu Ile 165 170 175 His Glu Asn Leu Pro Lys Phe Ile Asp Asn Ile Lys Ile Phe Glu Lys 180 185 190 Met Lys Lys Glu Ala Pro Glu Leu Leu Ser Pro Phe Asn Gln Thr Leu 195 200 205 Lys Asp Met Lys Asp Val Ile Lys Gly Thr Thr Leu Glu Glu Ile Phe 210 215 220 Ser Leu Asp Tyr Phe Asn Lys Thr Leu Thr Gln Ser Gly Ile Asp Ile 225 230 235 240 Tyr Asn Ser Val Ile Gly Gly Arg Thr Pro Glu Glu Gly Lys Thr Lys 245 250 255 Ile Lys Gly Leu Asn Glu Tyr Ile Asn Thr Asp Phe Asn Gln Lys Gln 260 265 270 Thr Asp Lys Lys Lys Arg Gln Pro Lys Phe Lys Gln Leu Tyr Lys Gln 275 280 285 Ile Leu Ser Asp Arg Gln Ser Leu Ser Phe Ile Ala Glu Ala Phe Lys 290 295 300 Asn Asp Thr Glu Ile Leu Glu Ala Ile Glu Lys Phe Tyr Val Asn Glu 305 310 315 320 Leu Leu His Phe Ser Asn Glu Gly Lys Ser Thr Asn Val Leu Asp Ala 325 330 335 Ile Lys Asn Ala Val Ser Asn Leu Glu Ser Phe Asn Leu Thr Lys Met 340 345 350 Tyr Phe Arg Ser Gly Ala Ser Leu Thr Asp Val Ser Arg Lys Val Phe 355 360 365 Gly Glu Trp Ser Ile Ile Asn Arg Ala Leu Asp Asn Tyr Tyr Ala Thr 370 375 380 Thr Tyr Pro Ile Lys Pro Arg Glu Lys Ser Glu Lys Tyr Glu Glu Arg 385 390 395 400 Lys Glu Lys Trp Leu Lys Gln Asp Phe Asn Val Ser Leu Ile Gln Thr 405 410 415 Ala Ile Asp Glu Tyr Asp Asn Glu Thr Val Lys Gly Lys Asn Ser Gly 420 425 430 Lys Val Ile Ala Asp Tyr Phe Ala Lys Phe Cys Asp Asp Lys Glu Thr 435 440 445 Asp Leu Ile Gln Lys Val Asn Glu Gly Tyr Ile Ala Val Lys Asp Leu 450 455 460 Leu Asn Thr Pro Cys Pro Glu Asn Glu Lys Leu Gly Ser Asn Lys Asp 465 470 475 480 Gln Val Lys Gln Ile Lys Ala Phe Met Asp Ser Ile Met Asp Ile Met 485 490 495 His Phe Val Arg Pro Leu Ser Leu Lys Asp Thr Asp Lys Glu Lys Asp 500 505 510 Glu Thr Phe Tyr Ser Leu Phe Thr Pro Leu Tyr Asp His Leu Thr Gln 515 520 525 Thr Ile Ala Leu Tyr Asn Lys Val Arg Asn Tyr Leu Thr Gln Lys Pro 530 535 540 Tyr Ser Thr Glu Lys Ile Lys Leu Asn Phe Glu Asn Ser Thr Leu Leu 545 550 555 560 Gly Gly Trp Asp Leu Asn Lys Glu Thr Asp Asn Thr Ala Ile Ile Leu 565 570 575 Arg Lys Asp Asn Leu Tyr Tyr Leu Gly Ile Met Asp Lys Arg His Asn 580 585 590 Arg Ile Phe Arg Asn Val Pro Lys Ala Asp Lys Lys Asp Phe Cys Tyr 595 600 605 Glu Lys Met Val Tyr Lys Leu Leu Pro Gly Ala Asn Lys Met Leu Pro 610 615 620 Lys Val Phe Phe Ser Gln Ser Arg Ile Gln Glu Phe Thr Pro Ser Ala 625 630 635 640 Lys Leu Leu Glu Asn Tyr Ala Asn Glu Thr His Lys Lys Gly Asp Asn 645 650 655 Phe Asn Leu Asn His Cys His Lys Leu Ile Asp Phe Phe Lys Asp Ser 660 665 670 Ile Asn Lys His Glu Asp Trp Lys Asn Phe Asp Phe Arg Phe Ser Ala 675 680 685 Thr Ser Thr Tyr Ala Asp Leu Ser Gly Phe Tyr His Glu Val Glu His 690 695 700 Gln Gly Tyr Lys Ile Ser Phe Gln Ser Val Ala Asp Ser Phe Ile Asp 705 710 715 720 Asp Leu Val Asn Glu Gly Lys Leu Tyr Leu Phe Gln Ile Tyr Asn Lys 725 730 735 Asp Phe Ser Pro Phe Ser Lys Gly Lys Pro Asn Leu His Thr Leu Tyr 740 745 750 Trp Lys Met Leu Phe Asp Glu Asn Asn Leu Lys Asp Val Val Tyr Lys 755 760 765 Leu Asn Gly Glu Ala Glu Val Phe Tyr Arg Lys Lys Ser Ile Ala Glu 770 775 780 Lys Asn Thr Thr Ile His Lys Ala Asn Glu Ser Ile Ile Asn Lys Asn 785 790 795 800 Pro Asp Asn Pro Lys Ala Thr Ser Thr Phe Asn Tyr Asp Ile Val Lys 805 810 815 Asp Lys Arg Tyr Thr Ile Asp Lys Phe Gln Phe His Ile Pro Ile Thr 820 825 830 Met Asn Phe Lys Ala Glu Gly Ile Phe Asn Met Asn Gln Arg Val Asn 835 840 845 Gln Phe Leu Lys Ala Asn Pro Asp Ile Asn Ile Ile Gly Ile Asp Arg 850 855 860 Gly Glu Arg His Leu Leu Tyr Tyr Ala Leu Ile Asn Gln Lys Gly Lys 865 870 875 880 Ile Leu Lys Gln Asp Thr Leu Asn Val Ile Ala Asn Glu Lys Gln Lys 885 890 895 Val Asp Tyr His Asn Leu Leu Asp Lys Lys Glu Gly Asp Arg Ala Thr 900 905 910 Ala Arg Gln Glu Trp Gly Val Ile Glu Thr Ile Lys Glu Leu Lys Glu 915 920 925 Gly Tyr Leu Ser Gln Val Ile His Lys Leu Thr Asp Leu Met Ile Glu 930 935 940 Asn Asn Ala Ile Ile Val Met Glu Asp Leu Asn Phe Gly Phe Lys Arg 945 950 955 960 Gly Arg Gln Lys Val Glu Lys Gln Val Tyr Gln Lys Phe Glu Lys Met 965 970 975 Leu Ile Asp Lys Leu Asn Tyr Leu Val Asp Lys Asn Lys Lys Ala Asn 980 985 990 Glu Leu Gly Gly Leu Leu Asn Ala Phe Gln Leu Ala Asn Lys Phe Glu 995 1000 1005 Ser Phe Gln Lys Met Gly Lys Gln Asn Gly Phe Ile Phe Tyr Val 1010 1015 1020 Pro Ala Trp Asn Thr Ser Lys Thr Asp Pro Ala Thr Gly Phe Ile 1025 1030 1035 Asp Phe Leu Lys Pro Arg Tyr Glu Asn Leu Asn Gln Ala Lys Asp 1040 1045 1050 Phe Phe Glu Lys Phe Asp Ser Ile Arg Leu Asn Ser Lys Ala Asp 1055 1060 1065 Tyr Phe Glu Phe Ala Phe Asp Phe Lys Asn Phe Thr Glu Lys Ala 1070 1075 1080 Asp Gly Gly Arg Thr Lys Trp Thr Val Cys Thr Thr Asn Glu Asp 1085 1090 1095 Arg Tyr Gln Trp Asn Arg Ala Leu Asn Asn Asn Arg Gly Ser Gln 1100 1105 1110 Glu Lys Tyr Asp Ile Thr Ala Glu Leu Lys Ser Leu Phe Asp Gly 1115 1120 1125 Lys Val Asp Tyr Lys Ser Gly Lys Asp Leu Lys Gln Gln Ile Ala 1130 1135 1140 Ser Gln Glu Ser Ala Asp Phe Phe Lys Ala Leu Met Lys Asn Leu 1145 1150 1155 Ser Ile Thr Leu Ser Leu Arg His Asn Asn Gly Glu Lys Gly Asp 1160 1165 1170 Asn Glu Gln Asp Tyr Ile Leu Ser Pro Val Ala Asp Ser Lys Gly 1175 1180 1185 Arg Phe Phe Asp Ser Arg Lys Ala Asp Asp Asp Met Pro Lys Asn 1190 1195 1200 Ala Asp Ala Asn Gly Ala Tyr His Ile Ala Leu Lys Gly Leu Trp 1205 1210 1215 Cys Leu Glu Gln Ile Ser Lys Thr Asp Asp Leu Lys Lys Val Lys 1220 1225 1230 Leu Ala Ile Ser Asn Lys Glu Trp Leu Glu Phe Val Gln Thr Leu 1235 1240 1245 Lys Gly 1250 <210> 46 <211> 228 <212> PRT <213> Rattus norvegicus <400> 46 Ser Ser Glu Thr Gly Pro Val Ala Val Asp Pro Thr Leu Arg Arg Arg 1 5 10 15 Ile Glu Pro His Glu Phe Glu Val Phe Phe Asp Pro Arg Glu Leu Arg 20 25 30 Lys Glu Thr Cys Leu Leu Tyr Glu Ile Asn Trp Gly Gly Arg His Ser 35 40 45 Ile Trp Arg His Thr Ser Gln Asn Thr Asn Lys His Val Glu Val Asn 50 55 60 Phe Ile Glu Lys Phe Thr Thr Glu Arg Tyr Phe Cys Pro Asn Thr Arg 65 70 75 80 Cys Ser Ile Thr Trp Phe Leu Ser Trp Ser Pro Cys Gly Glu Cys Ser 85 90 95 Arg Ala Ile Thr Glu Phe Leu Ser Arg Tyr Pro His Val Thr Leu Phe 100 105 110 Ile Tyr Ile Ala Arg Leu Tyr His His Ala Asp Pro Arg Asn Arg Gln 115 120 125 Gly Leu Arg Asp Leu Ile Ser Ser Gly Val Thr Ile Gln Ile Met Thr 130 135 140 Glu Gln Glu Ser Gly Tyr Cys Trp Arg Asn Phe Val Asn Tyr Ser Pro 145 150 155 160 Ser Asn Glu Ala His Trp Pro Arg Tyr Pro His Leu Trp Val Arg Leu 165 170 175 Tyr Val Leu Glu Leu Tyr Cys Ile Ile Leu Gly Leu Pro Pro Cys Leu 180 185 190 Asn Ile Leu Arg Arg Lys Gln Pro Gln Leu Thr Phe Phe Thr Ile Ala 195 200 205 Leu Gln Ser Cys His Tyr Gln Arg Leu Pro Pro His Ile Leu Trp Ala 210 215 220 Thr Gly Leu Lys 225 <210> 47 <211> 199 <212> PRT <213> Homo sapiens <400> 47 Met Glu Ala Ser Pro Ala Ser Gly Pro Arg His Leu Met Asp Pro His 1 5 10 15 Ile Phe Thr Ser Asn Phe Asn Asn Gly Ile Gly Arg His Lys Thr Tyr 20 25 30 Leu Cys Tyr Glu Val Glu Arg Leu Asp Asn Gly Thr Ser Val Lys Met 35 40 45 Asp Gln His Arg Gly Phe Leu His Asn Gln Ala Lys Asn Leu Leu Cys 50 55 60 Gly Phe Tyr Gly Arg His Ala Glu Leu Arg Phe Leu Asp Leu Val Pro 65 70 75 80 Ser Leu Gln Leu Asp Pro Ala Gln Ile Tyr Arg Val Thr Trp Phe Ile 85 90 95 Ser Trp Ser Pro Cys Phe Ser Trp Gly Cys Ala Gly Glu Val Arg Ala 100 105 110 Phe Leu Gln Glu Asn Thr His Val Arg Leu Arg Ile Phe Ala Ala Arg 115 120 125 Ile Tyr Asp Tyr Asp Pro Leu Tyr Lys Glu Ala Leu Gln Met Leu Arg 130 135 140 Asp Ala Gly Ala Gln Val Ser Ile Met Thr Tyr Asp Glu Phe Lys His 145 150 155 160 Cys Trp Asp Thr Phe Val Asp His Gln Gly Cys Pro Phe Gln Pro Trp 165 170 175 Asp Gly Leu Asp Glu His Ser Gln Ala Leu Ser Gly Arg Leu Arg Ala 180 185 190 Ile Leu Gln Asn Gln Gly Asn 195 <210> 48 <211> 83 <212> PRT <213> Bacteriophage <400> 48 Thr Asn Leu Ser Asp Ile Ile Glu Lys Glu Thr Gly Lys Gln Leu Val 1 5 10 15 Ile Gln Glu Ser Ile Leu Met Leu Pro Glu Glu Val Glu Glu Val Ile 20 25 30 Gly Asn Lys Pro Glu Ser Asp Ile Leu Val His Thr Ala Tyr Asp Glu 35 40 45 Ser Thr Asp Glu Asn Val Met Leu Leu Thr Ser Asp Ala Pro Glu Tyr 50 55 60 Lys Pro Trp Ala Leu Val Ile Gln Asp Ser Asn Gly Glu Asn Lys Ile 65 70 75 80 Lys Met Leu <210> 49 <211> 1596 <212> PRT <213> Artificial <220> <223> Base editor fusion protein <400> 49 Met Lys Arg Thr Ala Asp Gly Ser Glu Phe Glu Ser Pro Lys Lys Lys 1 5 10 15 Arg Lys Val Thr Asn Leu Ser Asp Ile Ile Glu Lys Glu Thr Gly Lys 20 25 30 Gln Leu Val Ile Gln Glu Ser Ile Leu Met Leu Pro Glu Glu Val Glu 35 40 45 Glu Val Ile Gly Asn Lys Pro Glu Ser Asp Ile Leu Val His Thr Ala 50 55 60 Tyr Asp Glu Ser Thr Asp Glu Asn Val Met Leu Leu Thr Ser Asp Ala 65 70 75 80 Pro Glu Tyr Lys Pro Trp Ala Leu Val Ile Gln Asp Ser Asn Gly Glu 85 90 95 Asn Lys Ile Lys Met Leu Ser Gly Gly Ser Gly Gly Ser Gly Gly Ser 100 105 110 Ser Lys Leu Glu Lys Phe Thr Asn Cys Tyr Ser Leu Ser Lys Thr Leu 115 120 125 Arg Phe Lys Ala Ile Pro Val Gly Lys Thr Gln Glu Asn Ile Asp Asn 130 135 140 Lys Arg Leu Leu Val Glu Asp Glu Lys Arg Ala Glu Asp Tyr Lys Gly 145 150 155 160 Val Lys Lys Leu Leu Asp Arg Tyr Tyr Leu Ser Phe Ile Asn Asp Val 165 170 175 Leucine Histidine Serine Isoleucine Lysine Leucine Lysine Asparagine Leucine Asparagine Asparagine Tyrosine Isoleucine Serine Leucine Phenylalanine 180 185 190 Arginine Lysine Lysine Threonine Arginine Threonine Glutamic acid Lysine Glutamic acid Asparagine Lysine Glutamic acid Leucine Glutamic acid Asparagine Leucine 195 200 205 Glutamic acid Isoleucine Asparagine Leucine Arginine Lysine Glutamic acid Isoleucine Alanine Lysine Alanine Phenylalanine Lysine Glycine Asparagine Glutamic acid 210 215 220 Glycine Tyrosine Lysine Serine Leucine Phenylalanine Lysine Lysine Aspartic acid Isoleucine Isoleucine Glutamic acid Threonine Isoleucine Leucine Proline 225 230 235 240 Glutamic acid Phenylalanine Leucine Aspartic acid Aspartic acid Lysine Aspartic acid Glutamic acid Isoleucine Alanine Leucine Valine Asparagine Serine Phenylalanine Asparagine 245 250 255 Glycine Phenylalanine Threonine Threonine Alanine Phenylalanine Threonine Glycine Phenylalanine Phenylalanine Aspartic acid Asparagine Arginine Glutamic acid Asparagine Methionine 260 265 270 Phenylalanine Serine Glutamic acid Glutamic acid Alanine Lysine Serine Threonine Serine Isoleucine Alanine Phenylalanine Arginine Cysteine Isoleucine Asparagine 275 280 285 Glutamic acid Asparagine Leucine Threonine Arginine Tyrosine Isoleucine Serine Asparagine Methionine Aspartic acid Isoleucine Phenylalanine Glutamic acid Lysine Valine 290 295 300 Aspartic acid Alanine Isoleucine Phenylalanine Aspartic acid Lysine Histidine Glutamic acid Valine Glutamine Glutamic acid Isoleucine Lysine Glutamic acid Lysine Isoleucine 305 310 315 320 Leucine Asparagine Serine Aspartic acid Tyrosine Aspartic acid Valine Glutamic acid Aspartic acid Phenylalanine Phenylalanine Glutamic acid Glycine Glutamic acid Phenylalanine Phenylalanine 325 330 335 Asn Phe Val Leu Thr Gln Glu Gly Ile Asp Val Tyr Asn Ala Ile Ile 340 345 350 Gly Gly Phe Val Thr Glu Ser Gly Glu Lys Ile Lys Gly Leu Asn Glu 355 360 365 Tyr Ile Asn Leu Tyr Asn Gln Lys Thr Lys Gln Lys Leu Pro Lys Phe 370 375 380 Lys Pro Leu Tyr Lys Gln Val Leu Ser Asp Arg Glu Ser Leu Ser Phe 385 390 395 400 Tyr Gly Glu Gly Tyr Thr Ser Asp Glu Glu Val Leu Glu Val Phe Arg 405 410 415 Asn Thr Leu Asn Lys Asn Ser Glu Ile Phe Ser Ser Ile Lys Lys Leu 420 425 430 Glu Lys Leu Phe Lys Asn Phe Asp Glu Tyr Ser Ser Ala Gly Ile Phe 435 440 445 Val Lys Asn Gly Pro Ala Ile Ser Thr Ile Ser Lys Asp Ile Phe Gly 450 455 460 Glu Trp Asn Val Ile Arg Asp Lys Trp Asn Ala Glu Tyr Asp Asp Ile 465 470 475 480 His Leu Lys Lys Lys Ala Val Val Thr Glu Lys Tyr Glu Asp Asp Arg 485 490 495 Arg Lys Ser Phe Lys Lys Ile Gly Ser Phe Ser Leu Glu Gln Leu Gln 500 505 510 Glu Tyr Ala Asp Ala Asp Leu Ser Val Val Glu Lys Leu Lys Glu Ile 515 520 525 Ile Ile Gln Lys Val Asp Glu Ile Tyr Lys Val Tyr Gly Ser Ser Glu 530 535 540 Lys Leu Phe Asp Ala Asp Phe Val Leu Glu Lys Ser Leu Lys Lys Asn 545 550 555 560 Asp Ala Val Val Ala Ile Met Lys Asp Leu Leu Asp Ser Val Lys Ser 565 570 575 Phe Glu Asn Tyr Ile Lys Ala Phe Phe Gly Glu Gly Lys Glu Thr Asn 580 585 590 Arg Asp Glu Ser Phe Tyr Gly Asp Phe Val Leu Ala Tyr Asp Ile Leu 595 600 605 Leu Lys Val Asp His Ile Tyr Asp Ala Ile Arg Asn Tyr Val Thr Gln 610 615 620 Lys Pro Tyr Ser Lys Asp Lys Phe Lys Leu Tyr Phe Gln Asn Pro Gln 625 630 635 640 Phe Met Gly Gly Trp Asp Lys Asp Lys Glu Thr Asp Tyr Arg Ala Thr 645 650 655 Ile Leu Arg Tyr Gly Ser Lys Tyr Tyr Leu Ala Ile Met Asp Lys Lys 660 665 670 Tyr Ala Lys Cys Leu Gln Lys Ile Asp Lys Asp Asp Val Asn Gly Asn 675 680 685 Tyr Glu Lys Ile Asn Tyr Lys Leu Leu Pro Gly Pro Asn Lys Met Leu 690 695 700 Pro Lys Val Phe Phe Ser Lys Lys Trp Met Ala Tyr Tyr Asn Pro Ser 705 710 715 720 Glu Asp Ile Gln Lys Ile Tyr Lys Asn Gly Thr Phe Lys Lys Gly Asp 725 730 735 Met Phe Asn Leu Asn Asp Cys His Lys Leu Ile Asp Phe Phe Lys Asp 740 745 750 Ser Ile Ser Arg Tyr Pro Lys Trp Ser Asn Ala Tyr Asp Phe Asn Phe 755 760 765 Ser Glu Thr Glu Lys Tyr Lys Asp Ile Ala Gly Phe Tyr Arg Glu Val 770 775 780 Glu Glu Gln Gly Tyr Lys Val Ser Phe Glu Ser Ala Ser Lys Lys Glu 785 790 795 800 Val Asp Lys Leu Val Glu Glu Gly Lys Leu Tyr Met Phe Gln Ile Tyr 805 810 815 Asn Lys Asp Phe Ser Asp Lys Ser His Gly Thr Pro Asn Leu His Thr 820 825 830 Met Tyr Phe Lys Leu Leu Phe Asp Glu Asn Asn His Gly Gln Ile Arg 835 840 845 Leu Ser Gly Gly Ala Glu Leu Phe Met Arg Arg Ala Ser Leu Lys Lys 850 855 860 Glu Glu Leu Val Val His Pro Ala Asn Ser Pro Ile Ala Asn Lys Asn 865 870 875 880 Pro Asp Asn Pro Lys Lys Thr Thr Thr Leu Ser Tyr Asp Val Tyr Lys 885 890 895 Asp Lys Arg Phe Ser Glu Asp Gln Tyr Glu Leu His Ile Pro Ile Ala 900 905 910 Ile Asn Lys Cys Pro Lys Asn Ile Phe Lys Ile Asn Thr Glu Val Arg 915 920 925 Val Leu Leu Lys His Asp Asp Asn Pro Tyr Val Ile Gly Ile Ala Arg 930 935 940 Gly Glu Arg Asn Leu Leu Tyr Ile Val Val Val Asp Gly Lys Gly Asn 945 950 955 960 Ile Val Glu Gln Tyr Ser Leu Asn Glu Ile Ile Asn Asn Phe Asn Gly 965 970 975 Ile Arg Ile Lys Thr Asp Tyr His Ser Leu Leu Asp Lys Lys Glu Lys 980 985 990 Glu Arg Phe Glu Ala Arg Gln Asn Trp Thr Ser Ile Glu Asn Ile Lys 995 1000 1005 Glu Leu Lys Ala Gly Tyr Ile Ser Gln Val Val His Lys Ile Cys 1010 1015 1020 Glu Leu Val Glu Lys Tyr Asp Ala Val Ile Ala Leu Glu Asp Leu 1025 1030 1035 Asn Ser Gly Phe Lys Asn Ser Arg Val Lys Val Glu Lys Gln Val 1040 1045 1050 Tyr Gln Lys Phe Glu Lys Met Leu Ile Asp Lys Leu Asn Tyr Met 1055 1060 1065 Val Asp Lys Lys Ser Asn Pro Cys Ala Thr Gly Gly Ala Leu Lys 1070 1075 1080 Gly Tyr Gln Ile Thr Asn Lys Phe Glu Ser Phe Lys Ser Met Ser 1085 1090 1095 Thr Gln Asn Gly Phe Ile Phe Tyr Ile Pro Ala Trp Leu Thr Ser 1100 1105 1110 Lys Ile Asp Pro Ser Thr Gly Phe Val Asn Leu Leu Lys Thr Lys 1115 1120 1125 Tyr Thr Ser Ile Ala Asp Ser Lys Lys Phe Ile Ser Ser Phe Asp 1130 1135 1140 Arg Ile Met Tyr Val Pro Glu Glu Asp Leu Phe Glu Phe Ala Leu 1145 1150 1155 Asp Tyr Lys Asn Phe Ser Arg Thr Asp Ala Asp Tyr Ile Lys Lys 1160 1165 1170 Trp Lys Leu Tyr Ser Tyr Gly Asn Arg Ile Arg Ile Phe Arg Asn 1175 1180 1185 Pro Lys Lys Asn Asn Val Phe Asp Trp Glu Glu Val Cys Leu Thr 1190 1195 1200 Ser Ala Tyr Lys Glu Leu Phe Asn Lys Tyr Gly Ile Asn Tyr Gln 1205 1210 1215 Gln Gly Asp Ile Arg Ala Leu Leu Cys Glu Gln Ser Asp Lys Ala 1220 1225 1230 Phe Tyr Ser Ser Phe Met Ala Leu Met Ser Leu Met Leu Gln Met 1235 1240 1245 Arg Asn Ser Ile Thr Gly Arg Thr Asp Val Asp Phe Leu Ile Ser 1250 1255 1260 Pro Val Lys Asn Ser Asp Gly Ile Phe Tyr Asp Ser Arg Asn Tyr 1265 1270 1275 Glu Ala Gln Glu Asn Ala Ile Leu Pro Lys Asn Ala Asp Ala Asn 1280 1285 1290 Gly Ala Tyr Asn Ile Ala Arg Lys Val Leu Trp Ala Ile Gly Gln 1295 1300 1305 Phe Lys Lys Ala Glu Asp Glu Lys Leu Asp Lys Val Lys Ile Ala 1310 1315 1320 Ile Ser Asn Lys Glu Trp Leu Glu Tyr Ala Gln Thr Ser Val Lys 1325 1330 1335 His Asp Ser Pro Glu Ile Pro Gly Glu Pro Pro Pro Pro Ile Pro 1340 1345 1350 Pro Glu Ser Ser Glu Thr Gly Pro Val Ala Val Asp Pro Thr Leu 1355 1360 1365 Arg Arg Arg Ile Glu Pro His Glu Phe Glu Val Phe Phe Asp Pro 1370 1375 1380 Arg Glu Leu Arg Lys Glu Thr Cys Leu Leu Tyr Glu Ile Asn Trp 1385 1390 1395 Gly Gly Arg His Ser Ile Trp Arg His Thr Ser Gln Asn Thr Asn 1400 1405 1410 Lys His Val Glu Val Asn Phe Ile Glu Lys Phe Thr Thr Glu Arg 1415 1420 1425 Tyr Phe Cys Pro Asn Thr Arg Cys Ser Ile Thr Trp Phe Leu Ser 1430 1435 1440 Trp Ser Pro Cys Gly Glu Cys Ser Arg Ala Ile Thr Glu Phe Leu 1445 1450 1455 Ser Arg Tyr Pro His Val Thr Leu Phe Ile Tyr Ile Ala Arg Leu 1460 1465 1470 Tyr His His Ala Asp Pro Arg Asn Arg Gln Gly Leu Arg Asp Leu 1475 1480 1485 Ile Ser Ser Gly Val Thr Ile Gln Ile Met Thr Glu Gln Glu Ser 1490 1495 1500 Gly Tyr Cys Trp Arg Asn Phe Val Asn Tyr Ser Pro Ser Asn Glu 1505 1510 1515 Ala His Trp Pro Arg Tyr Pro His Leu Trp Val Arg Leu Tyr Val 1520 1525 1530 Leu Glu Leu Tyr Cys Ile Ile Leu Gly Leu Pro Pro Cys Leu Asn 1535 1540 1545 Ile Leu Arg Arg Lys Gln Pro Gln Leu Thr Phe Phe Thr Ile Ala 1550 1555 1560 Leu Gln Ser Cys His Tyr Gln Arg Leu Pro Pro His Ile Leu Trp 1565 1570 1575 Ala Thr Gly Leu Lys Ser Gly Gly Ser Lys Arg Thr Ala Asp Gly 1580 1585 1590 Ser Glu Phe 1595 <210> 50 <211> 1594 <212> PRT <213> Artificial <220> <223> Base editor fusion protein <400> 50 Met Lys Arg Thr Ala Asp Gly Ser Glu Phe Glu Ser Pro Lys Lys Lys 1 5 10 15 Arg Lys Val Thr Asn Leu Ser Asp Ile Ile Glu Lys Glu Thr Gly Lys 20 25 30 Gln Leu Val Ile Gln Glu Ser Ile Leu Met Leu Pro Glu Glu Val Glu 35 40 45 Glu Val Ile Gly Asn Lys Pro Glu Ser Asp Ile Leu Val His Thr Ala 50 55 60 Tyr Asp Glu Ser Thr Asp Glu Asn Val Met Leu Leu Thr Ser Asp Ala 65 70 75 80 Pro Glu Tyr Lys Pro Trp Ala Leu Val Ile Gln Asp Ser Asn Gly Glu 85 90 95 Asn Lys Ile Lys Met Leu Ser Gly Gly Ser Gly Gly Ser Gly Gly Ser 100 105 110 Ser Lys Leu Glu Lys Phe Thr Asn Cys Tyr Ser Leu Ser Lys Thr Leu 115 120 125 Arg Phe Lys Ala Ile Pro Val Gly Lys Thr Gln Glu Asn Ile Asp Asn 130 135 140 Lys Arg Leu Leu Val Glu Asp Glu Lys Arg Ala Glu Asp Tyr Lys Gly 145 150 155 160 Val Lys Lys Leu Leu Asp Arg Tyr Tyr Leu Ser Phe Ile Asn Asp Val 165 170 175 Leu His Ser Ile Lys Leu Lys Asn Leu Asn Asn Tyr Ile Ser Leu Phe 180 185 190 Arg Lys Lys Thr Arg Thr Glu Lys Glu Asn Lys Glu Leu Glu Asn Leu 195 200 205 Glu Ile Asn Leu Arg Lys Glu Ile Ala Lys Ala Phe Lys Gly Asn Glu 210 215 220 Gly Tyr Lys Ser Leu Phe Lys Lys Asp Ile Ile Glu Thr Ile Leu Pro 225 230 235 240 Glu Phe Leu Asp Asp Lys Asp Glu Ile Ala Leu Val Asn Ser Phe Asn 245 250 255 Gly Phe Thr Thr Ala Phe Thr Gly Phe Phe Asp Asn Arg Glu Asn Met 260 265 270 Phe Ser Glu Glu Ala Lys Ser Thr Ser Ile Ala Phe Arg Cys Ile Asn 275 280 285 Glu Asn Leu Thr Arg Tyr Ile Ser Asn Met Asp Ile Phe Glu Lys Val 290 295 300 Asp Ala Ile Phe Asp Lys His Glu Val Gln Glu Ile Lys Glu Lys Ile 305 310 315 320 Leu Asn Ser Asp Tyr Asp Val Glu Asp Phe Phe Glu Gly Glu Phe Phe 325 330 335 Asn Phe Val Leu Thr Gln Glu Gly Ile Asp Val Tyr Asn Ala Ile Ile 340 345 350 Gly Gly Phe Val Thr Glu Ser Gly Glu Lys Ile Lys Gly Leu Asn Glu 355 360 365 Tyr Ile Asn Leu Tyr Asn Gln Lys Thr Lys Gln Lys Leu Pro Lys Phe 370 375 380 Lys Pro Leu Tyr Lys Gln Val Leu Ser Asp Arg Glu Ser Leu Ser Phe 385 390 395 400 Tyr Gly Glu Gly Tyr Thr Ser Asp Glu Glu Val Leu Glu Val Phe Arg 405 410 415 Asn Thr Leu Asn Lys Asn Ser Glu Ile Phe Ser Ser Ile Lys Lys Leu 420 425 430 Glu Lys Leu Phe Lys Asn Phe Asp Glu Tyr Ser Ser Ala Gly Ile Phe 435 440 445 Val Lys Asn Gly Pro Ala Ile Ser Thr Ile Ser Lys Asp Ile Phe Gly 450 455 460 Glu Trp Asn Val Ile Arg Asp Lys Trp Asn Ala Glu Tyr Asp Asp Ile 465 470 475 480 His Leu Lys Lys Lys Ala Val Val Thr Glu Lys Tyr Glu Asp Asp Arg 485 490 495 Arg Lys Ser Phe Lys Lys Ile Gly Ser Phe Ser Leu Glu Gln Leu Gln 500 505 510 Glu Tyr Ala Asp Ala Asp Leu Ser Val Val Glu Lys Leu Lys Glu Ile 515 520 525 Ile Ile Gln Lys Val Asp Glu Ile Tyr Lys Val Tyr Gly Ser Ser Glu 530 535 540 Lys Leu Phe Asp Ala Asp Phe Val Leu Glu Lys Ser Leu Lys Lys Asn 545 550 555 560 Asp Ala Val Val Ala Ile Met Lys Asp Leu Leu Asp Ser Val Lys Ser 565 570 575 Phe Glu Asn Tyr Ile Lys Ala Phe Phe Gly Glu Gly Lys Glu Thr Asn 580 585 590 Arg Asp Glu Ser Phe Tyr Gly Asp Phe Val Leu Ala Tyr Asp Ile Leu 595 600 605 Leu Lys Val Asp His Ile Tyr Asp Ala Ile Arg Asn Tyr Val Thr Gln 610 615 620 Lys Pro Tyr Ser Lys Asp Lys Phe Lys Leu Tyr Phe Gln Asn Pro Gln 625 630 635 640 Phe Met Gly Gly Trp Asp Lys Asp Lys Glu Thr Asp Tyr Arg Ala Thr 645 650 655 Ile Leu Arg Tyr Gly Ser Lys Tyr Tyr Leu Ala Ile Met Asp Lys Lys 660 665 670 Tyr Ala Lys Cys Leu Gln Lys Ile Asp Lys Asp Asp Val Asn Gly Asn 675 680 685 Tyr Glu Lys Ile Asn Tyr Lys Leu Leu Pro Gly Pro Asn Lys Met Leu 690 695 700 Pro Lys Val Phe Phe Ser Lys Lys Trp Met Ala Tyr Tyr Asn Pro Ser 705 710 715 720 Glu Asp Ile Gln Lys Ile Tyr Lys Asn Gly Thr Phe Lys Lys Gly Asp 725 730 735 Met Phe Asn Leu Asn Asp Cys His Lys Leu Ile Asp Phe Phe Lys Asp 740 745 750 Ser Ile Ser Arg Tyr Pro Lys Trp Ser Asn Ala Tyr Asp Phe Asn Phe 755 760 765 Ser Glu Thr Glu Lys Tyr Lys Asp Ile Ala Gly Phe Tyr Arg Glu Val 770 775 780 Glu Glu Gln Gly Tyr Lys Val Ser Phe Glu Ser Ala Ser Lys Lys Glu 785 790 795 800 Val Asp Lys Leu Val Glu Glu Gly Lys Leu Tyr Met Phe Gln Ile Tyr 805 810 815 Asn Lys Asp Phe Ser Asp Lys Ser His Gly Thr Pro Asn Leu His Thr 820 825 830 Met Tyr Phe Lys Leu Leu Phe Asp Glu Asn Asn His Gly Gln Ile Arg 835 840 845 Leu Ser Gly Gly Ala Glu Leu Phe Met Arg Arg Ala Ser Leu Lys Lys 850 855 860 Glu Glu Leu Val Val His Pro Ala Asn Ser Pro Ile Ala Asn Lys Asn 865 870 875 880 Pro Asp Asn Pro Lys Lys Thr Thr Thr Leu Ser Tyr Asp Val Tyr Lys 885 890 895 Asp Lys Arg Phe Ser Glu Asp Gln Tyr Glu Leu His Ile Pro Ile Ala 900 905 910 Ile Asn Lys Cys Pro Lys Asn Ile Phe Lys Ile Asn Thr Glu Val Arg 915 920 925 Val Leu Leu Lys His Asp Asp Asn Pro Tyr Val Ile Gly Ile Ala Arg 930 935 940 Gly Glu Arg Asn Leu Leu Tyr Ile Val Val Val Asp Gly Lys Gly Asn 945 950 955 960 Ile Val Glu Gln Tyr Ser Leu Asn Glu Ile Ile Asn Asn Phe Asn Gly 965 970 975 Ile Arg Ile Lys Thr Asp Tyr His Ser Leu Leu Asp Lys Lys Glu Lys 980 985 990 Glu Arg Phe Glu Ala Arg Gln Asn Trp Thr Ser Ile Glu Asn Ile Lys 995 1000 1005 Glu Leu Lys Ala Gly Tyr Ile Ser Gln Val Val His Lys Ile Cys 1010 1015 1020 Glu Leu Val Glu Lys Tyr Asp Ala Val Ile Ala Leu Glu Asp Leu 1025 1030 1035 Asn Ser Gly Phe Lys Asn Ser Arg Val Lys Val Glu Lys Gln Val 1040 1045 1050 Tyr Gln Lys Phe Glu Lys Met Leu Ile Asp Lys Leu Asn Tyr Met 1055 1060 1065 Val Asp Lys Lys Ser Asn Pro Cys Ala Thr Gly Gly Ala Leu Lys 1070 1075 1080 Gly Tyr Gln Ile Thr Asn Lys Phe Glu Ser Phe Lys Ser Met Ser 1085 1090 1095 Thr Gln Asn Gly Phe Ile Phe Tyr Ile Pro Ala Trp Leu Thr Ser 1100 1105 1110 Lys Ile Asp Pro Ser Thr Gly Phe Val Asn Leu Leu Lys Thr Lys 1115 1120 1125 Tyr Thr Ser Ile Ala Asp Ser Lys Lys Phe Ile Ser Ser Phe Asp 1130 1135 1140 Arg Ile Met Tyr Val Pro Glu Glu Asp Leu Phe Glu Phe Ala Leu 1145 1150 1155 Asp Tyr Lys Asn Phe Ser Arg Thr Asp Ala Asp Tyr Ile Lys Lys 1160 1165 1170 Trp Lys Leu Tyr Ser Tyr Gly Asn Arg Ile Arg Ile Phe Arg Asn 1175 1180 1185 Pro Lys Lys Asn Asn Val Phe Asp Trp Glu Glu Val Cys Leu Thr 1190 1195 1200 Ser Ala Tyr Lys Glu Leu Phe Asn Lys Tyr Gly Ile Asn Tyr Gln 1205 1210 1215 Gln Gly Asp Ile Arg Ala Leu Leu Cys Glu Gln Ser Asp Lys Ala 1220 1225 1230 Phe Tyr Ser Ser Phe Met Ala Leu Met Ser Leu Met Leu Gln Met 1235 1240 1245 Arg Asn Ser Ile Thr Gly Arg Thr Asp Val Asp Phe Leu Ile Ser 1250 1255 1260 Pro Val Lys Asn Ser Asp Gly Ile Phe Tyr Asp Ser Arg Asn Tyr 1265 1270 1275 Glu Ala Gln Glu Asn Ala Ile Leu Pro Lys Asn Ala Asp Ala Asn 1280 1285 1290 Gly Ala Tyr Asn Ile Ala Arg Lys Val Leu Trp Ala Ile Gly Gln 1295 1300 1305 Phe Lys Lys Ala Glu Asp Glu Lys Leu Asp Lys Val Lys Ile Ala 1310 1315 1320 Ile Ser Asn Lys Glu Trp Leu Glu Tyr Ala Gln Thr Ser Val Lys 1325 1330 1335 His Arg Pro Leu Pro His Asp Asn Asn Lys Gln Asp Tyr Ser Lys 1340 1345 1350 Ser Ser Glu Thr Gly Pro Val Ala Val Asp Pro Thr Leu Arg Arg 1355 1360 1365 Arg Ile Glu Pro His Glu Phe Glu Val Phe Phe Asp Pro Arg Glu 1370 1375 1380 Leu Arg Lys Glu Thr Cys Leu Leu Tyr Glu Ile Asn Trp Gly Gly 1385 1390 1395 Arg His Ser Ile Trp Arg His Thr Ser Gln Asn Thr Asn Lys His 1400 1405 1410 Val Glu Val Asn Phe Ile Glu Lys Phe Thr Thr Glu Arg Tyr Phe 1415 1420 1425 Cys Pro Asn Thr Arg Cys Ser Ile Thr Trp Phe Leu Ser Trp Ser 1430 1435 1440 Pro Cys Gly Glu Cys Ser Arg Ala Ile Thr Glu Phe Leu Ser Arg 1445 1450 1455 Tyr Pro His Val Thr Leu Phe Ile Tyr Ile Ala Arg Leu Tyr His 1460 1465 1470 His Ala Asp Pro Arg Asn Arg Gln Gly Leu Arg Asp Leu Ile Ser 1475 1480 1485 Ser Gly Val Thr Ile Gln Ile Met Thr Glu Gln Glu Ser Gly Tyr 1490 1495 1500 Cys Trp Arg Asn Phe Val Asn Tyr Ser Pro Ser Asn Glu Ala His 1505 1510 1515 Trp Pro Arg Tyr Pro His Leu Trp Val Arg Leu Tyr Val Leu Glu 1520 1525 1530 Leu Tyr Cys Ile Ile Leu Gly Leu Pro Pro Cys Leu Asn Ile Leu 1535 1540 1545 Arg Arg Lys Gln Pro Gln Leu Thr Phe Phe Thr Ile Ala Leu Gln 1550 1555 1560 Ser Cys His Tyr Gln Arg Leu Pro Pro His Ile Leu Trp Ala Thr 1565 1570 1575 Gly Leu Lys Ser Gly Gly Ser Lys Arg Thr Ala Asp Gly Ser Glu 1580 1585 1590 Phe <210> 51 <211> 1596 <212> PRT <213> Artificial <220> <223> Base editor fusion protein <400> 51 Met Lys Arg Thr Ala Asp Gly Ser Glu Phe Glu Ser Pro Lys Lys Lys 1 5 10 15 Arg Lys Val Thr Asn Leu Ser Asp Ile Ile Glu Lys Glu Thr Gly Lys 20 25 30 Gln Leu Val Ile Gln Glu Ser Ile Leu Met Leu Pro Glu Glu Val Glu 35 40 45 Glu Val Ile Gly Asn Lys Pro Glu Ser Asp Ile Leu Val His Thr Ala 50 55 60 Tyr Asp Glu Ser Thr Asp Glu Asn Val Met Leu Leu Thr Ser Asp Ala 65 70 75 80 Pro Glu Tyr Lys Pro Trp Ala Leu Val Ile Gln Asp Ser Asn Gly Glu 85 90 95 Asn Lys Ile Lys Met Leu Ser Gly Gly Ser Gly Gly Ser Gly Gly Ser 100 105 110 Ser Lys Leu Glu Lys Phe Thr Asn Cys Tyr Ser Leu Ser Lys Thr Leu 115 120 125 Arg Phe Lys Ala Ile Pro Val Gly Lys Thr Gln Glu Asn Ile Asp Asn 130 135 140 Lys Arg Leu Leu Val Glu Asp Glu Lys Arg Ala Glu Asp Tyr Lys Gly 145 150 155 160 Val Lys Lys Leu Leu Asp Arg Tyr Tyr Leu Ser Phe Ile Asn Asp Val 165 170 175 Leu His Ser Ile Lys Leu Lys Asn Leu Asn Asn Tyr Ile Ser Leu Phe 180 185 190 Arg Lys Lys Thr Arg Thr Glu Lys Glu Asn Lys Glu Leu Glu Asn Leu 195 200 205 Glu Ile Asn Leu Arg Lys Glu Ile Ala Lys Ala Phe Lys Gly Asn Glu 210 215 220 Gly Tyr Lys Ser Leu Phe Lys Lys Asp Ile Ile Glu Thr Ile Leu Pro 225 230 235 240 Glu Phe Leu Asp Asp Lys Asp Glu Ile Ala Leu Val Asn Ser Phe Asn 245 250 255 Gly Phe Thr Thr Ala Phe Thr Gly Phe Phe Asp Asn Arg Glu Asn Met 260 265 270 Phe Ser Glu Glu Ala Lys Ser Thr Ser Ile Ala Phe Arg Cys Ile Asn 275 280 285 Glu Asn Leu Thr Arg Tyr Ile Ser Asn Met Asp Ile Phe Glu Lys Val 290 295 300 Asp Ala Ile Phe Asp Lys His Glu Val Gln Glu Ile Lys Glu Lys Ile 305 310 315 320 Leu Asn Ser Asp Tyr Asp Val Glu Asp Phe Phe Glu Gly Glu Phe Phe 325 330 335 Asn Phe Val Leu Thr Gln Glu Gly Ile Asp Val Tyr Asn Ala Ile Ile 340 345 350 Gly Gly Phe Val Thr Glu Ser Gly Glu Lys Ile Lys Gly Leu Asn Glu 355 360 365 Tyr Ile Asn Leu Tyr Asn Gln Lys Thr Lys Gln Lys Leu Pro Lys Phe 370 375 380 Lys Pro Leu Tyr Lys Gln Val Leu Ser Asp Arg Glu Ser Leu Ser Phe 385 390 395 400 Tyr Gly Glu Gly Tyr Thr Ser Asp Glu Glu Val Leu Glu Val Phe Arg 405 410 415 Asn Thr Leu Asn Lys Asn Ser Glu Ile Phe Ser Ser Ile Lys Lys Leu 420 425 430 Glu Lys Leu Phe Lys Asn Phe Asp Glu Tyr Ser Ser Ala Gly Ile Phe 435 440 445 Val Lys Asn Gly Pro Ala Ile Ser Thr Ile Ser Lys Asp Ile Phe Gly 450 455 460 Glu Trp Asn Val Ile Arg Asp Lys Trp Asn Ala Glu Tyr Asp Asp Ile 465 470 475 480 His Leu Lys Lys Lys Ala Val Val Thr Glu Lys Tyr Glu Asp Asp Arg 485 490 495 Arg Lys Ser Phe Lys Lys Ile Gly Ser Phe Ser Leu Glu Gln Leu Gln 500 505 510 Glu Tyr Ala Asp Ala Asp Leu Ser Val Val Glu Lys Leu Lys Glu Ile 515 520 525 Ile Ile Gln Lys Val Asp Glu Ile Tyr Lys Val Tyr Gly Ser Ser Glu 530 535 540 Lys Leu Phe Asp Ala Asp Phe Val Leu Glu Lys Ser Leu Lys Lys Asn 545 550 555 560 Asp Ala Val Val Ala Ile Met Lys Asp Leu Leu Asp Ser Val Lys Ser 565 570 575 Phe Glu Asn Tyr Ile Lys Ala Phe Phe Gly Glu Gly Lys Glu Thr Asn 580 585 590 Arg Asp Glu Ser Phe Tyr Gly Asp Phe Val Leu Ala Tyr Asp Ile Leu 595 600 605 Leu Lys Val Asp His Ile Tyr Asp Ala Ile Arg Asn Tyr Val Thr Gln 610 615 620 Lys Pro Tyr Ser Lys Asp Lys Phe Lys Leu Tyr Phe Gln Asn Pro Gln 625 630 635 640 Phe Met Gly Gly Trp Asp Lys Asp Lys Glu Thr Asp Tyr Arg Ala Thr 645 650 655 Ile Leu Arg Tyr Gly Ser Lys Tyr Tyr Leu Ala Ile Met Asp Lys Lys 660 665 670 Tyr Ala Lys Cys Leu Gln Lys Ile Asp Lys Asp Asp Val Asn Gly Asn 675 680 685 Tyr Glu Lys Ile Asn Tyr Lys Leu Leu Pro Gly Pro Asn Lys Met Leu 690 695 700 Pro Lys Val Phe Phe Ser Lys Lys Trp Met Ala Tyr Tyr Asn Pro Ser 705 710 715 720 Glu Asp Ile Gln Lys Ile Tyr Lys Asn Gly Thr Phe Lys Lys Gly Asp 725 730 735 Met Phe Asn Leu Asn Asp Cys His Lys Leu Ile Asp Phe Phe Lys Asp 740 745 750 Ser Ile Ser Arg Tyr Pro Lys Trp Ser Asn Ala Tyr Asp Phe Asn Phe 755 760 765 Ser Glu Thr Glu Lys Tyr Lys Asp Ile Ala Gly Phe Tyr Arg Glu Val 770 775 780 Glu Glu Gln Gly Tyr Lys Val Ser Phe Glu Ser Ala Ser Lys Lys Glu 785 790 795 800 Val Asp Lys Leu Val Glu Glu Gly Lys Leu Tyr Met Phe Gln Ile Tyr 805 810 815 Asn Lys Asp Phe Ser Asp Lys Ser His Gly Thr Pro Asn Leu His Thr 820 825 830 Met Tyr Phe Lys Leu Leu Phe Asp Glu Asn Asn His Gly Gln Ile Arg 835 840 845 Leu Ser Gly Gly Ala Glu Leu Phe Met Arg Arg Ala Ser Leu Lys Lys 850 855 860 Glu Glu Leu Val Val His Pro Ala Asn Ser Pro Ile Ala Asn Lys Asn 865 870 875 880 Pro Asp Asn Pro Lys Lys Thr Thr Thr Leu Ser Tyr Asp Val Tyr Lys 885 890 895 Asp Lys Arg Phe Ser Glu Asp Gln Tyr Glu Leu His Ile Pro Ile Ala 900 905 910 Ile Asn Lys Cys Pro Lys Asn Ile Phe Lys Ile Asn Thr Glu Val Arg 915 920 925 Val Leu Leu Lys His Asp Asp Asn Pro Tyr Val Ile Gly Ile Ala Arg 930 935 940 Gly Glu Arg Asn Leu Leu Tyr Ile Val Val Val Asp Gly Lys Gly Asn 945 950 955 960 Ile Val Glu Gln Tyr Ser Leu Asn Glu Ile Ile Asn Asn Phe Asn Gly 965 970 975 Ile Arg Ile Lys Thr Asp Tyr His Ser Leu Leu Asp Lys Lys Glu Lys 980 985 990 Glu Arg Phe Glu Ala Arg Gln Asn Trp Thr Ser Ile Glu Asn Ile Lys 995 1000 1005 Glu Leu Lys Ala Gly Tyr Ile Ser Gln Val Val His Lys Ile Cys 1010 1015 1020 Glu Leu Val Glu Lys Tyr Asp Ala Val Ile Ala Leu Glu Asp Leu 1025 1030 1035 Asn Ser Gly Phe Lys Asn Ser Arg Val Lys Val Glu Lys Gln Val 1040 1045 1050 Tyr Gln Lys Phe Glu Lys Met Leu Ile Asp Lys Leu Asn Tyr Met 1055 1060 1065 Val Asp Lys Lys Ser Asn Pro Cys Ala Thr Gly Gly Ala Leu Lys 1070 1075 1080 Gly Tyr Gln Ile Thr Asn Lys Phe Glu Ser Phe Lys Ser Met Ser 1085 1090 1095 Thr Gln Asn Gly Phe Ile Phe Tyr Ile Pro Ala Trp Leu Thr Ser 1100 1105 1110 Lys Ile Asp Pro Ser Thr Gly Phe Val Asn Leu Leu Lys Thr Lys 1115 1120 1125 Tyr Thr Ser Ile Ala Asp Ser Lys Lys Phe Ile Ser Ser Phe Asp 1130 1135 1140 Arg Ile Met Tyr Val Pro Glu Glu Asp Leu Phe Glu Phe Ala Leu 1145 1150 1155 Asp Tyr Lys Asn Phe Ser Arg Thr Asp Ala Asp Tyr Ile Lys Lys 1160 1165 1170 Trp Lys Leu Tyr Ser Tyr Gly Asn Arg Ile Arg Ile Phe Arg Asn 1175 1180 1185 Pro Lys Lys Asn Asn Val Phe Asp Trp Glu Glu Val Cys Leu Thr 1190 1195 1200 Ser Ala Tyr Lys Glu Leu Phe Asn Lys Tyr Gly Ile Asn Tyr Gln 1205 1210 1215 Gln Gly Asp Ile Arg Ala Leu Leu Cys Glu Gln Ser Asp Lys Ala 1220 1225 1230 Phe Tyr Ser Ser Phe Met Ala Leu Met Ser Leu Met Leu Gln Met 1235 1240 1245 Arg Asn Ser Ile Thr Gly Arg Thr Asp Val Asp Phe Leu Ile Ser 1250 1255 1260 Pro Val Lys Asn Ser Asp Gly Ile Phe Tyr Asp Ser Arg Asn Tyr 1265 1270 1275 Glu Ala Gln Glu Asn Ala Ile Leu Pro Lys Asn Ala Asp Ala Asn 1280 1285 1290 Gly Ala Tyr Asn Ile Ala Arg Lys Val Leu Trp Ala Ile Gly Gln 1295 1300 1305 Phe Lys Lys Ala Glu Asp Glu Lys Leu Asp Lys Val Lys Ile Ala 1310 1315 1320 Ile Ser Asn Lys Glu Trp Leu Glu Tyr Ala Gln Thr Ser Val Lys 1325 1330 1335 His Gly Pro Leu Pro Ala Pro Pro Pro Gln Pro Pro Pro Pro Gln 1340 1345 1350 Pro Asn Ser Ser Glu Thr Gly Pro Val Ala Val Asp Pro Thr Leu 1355 1360 1365 Arg Arg Arg Ile Glu Pro His Glu Phe Glu Val Phe Phe Asp Pro 1370 1375 1380 Arg Glu Leu Arg Lys Glu Thr Cys Leu Leu Tyr Glu Ile Asn Trp 1385 1390 1395 Gly Gly Arg His Ser Ile Trp Arg His Thr Ser Gln Asn Thr Asn 1400 1405 1410 Lys His Val Glu Val Asn Phe Ile Glu Lys Phe Thr Thr Glu Arg 1415 1420 1425 Tyr Phe Cys Pro Asn Thr Arg Cys Ser Ile Thr Trp Phe Leu Ser 1430 1435 1440 Trp Ser Pro Cys Gly Glu Cys Ser Arg Ala Ile Thr Glu Phe Leu 1445 1450 1455 Ser Arg Tyr Pro His Val Thr Leu Phe Ile Tyr Ile Ala Arg Leu 1460 1465 1470 Tyr His His Ala Asp Pro Arg Asn Arg Gln Gly Leu Arg Asp Leu 1475 1480 1485 Ile Ser Ser Gly Val Thr Ile Gln Ile Met Thr Glu Gln Glu Ser 1490 1495 1500 Gly Tyr Cys Trp Arg Asn Phe Val Asn Tyr Ser Pro Ser Asn Glu 1505 1510 1515 Ala His Trp Pro Arg Tyr Pro His Leu Trp Val Arg Leu Tyr Val 1520 1525 1530 Leu Glu Leu Tyr Cys Ile Ile Leu Gly Leu Pro Pro Cys Leu Asn 1535 1540 1545 Ile Leu Arg Arg Lys Gln Pro Gln Leu Thr Phe Phe Thr Ile Ala 1550 1555 1560 Leu Gln Ser Cys His Tyr Gln Arg Leu Pro Pro His Ile Leu Trp 1565 1570 1575 Ala Thr Gly Leu Lys Ser Gly Gly Ser Lys Arg Thr Ala Asp Gly 1580 1585 1590 Ser Glu Phe 1595 <210> 52 <211> 1596 <212> PRT <213> Artificial <220> <223> Base editor fusion protein <400> 52 Met Lys Arg Thr Ala Asp Gly Ser Glu Phe Glu Ser Pro Lys Lys Lys 1 5 10 15 Arg Lys Val Thr Asn Leu Ser Asp Ile Ile Glu Lys Glu Thr Gly Lys 20 25 30 Gln Leu Val Ile Gln Glu Ser Ile Leu Met Leu Pro Glu Glu Val Glu 35 40 45 Glu Val Ile Gly Asn Lys Pro Glu Ser Asp Ile Leu Val His Thr Ala 50 55 60 Tyr Asp Glu Ser Thr Asp Glu Asn Val Met Leu Leu Thr Ser Asp Ala 65 70 75 80 Pro Glu Tyr Lys Pro Trp Ala Leu Val Ile Gln Asp Ser Asn Gly Glu 85 90 95 Asn Lys Ile Lys Met Leu Ser Gly Gly Ser Gly Gly Ser Gly Gly Ser 100 105 110 Ser Lys Leu Glu Lys Phe Thr Asn Cys Tyr Ser Leu Ser Lys Thr Leu 115 120 125 Arg Phe Lys Ala Ile Pro Val Gly Lys Thr Gln Glu Asn Ile Asp Asn 130 135 140 Lys Arg Leu Leu Val Glu Asp Glu Lys Arg Ala Glu Asp Tyr Lys Gly 145 150 155 160 Val Lys Lys Leu Leu Asp Arg Tyr Tyr Leu Ser Phe Ile Asn Asp Val 165 170 175 Leu His Ser Ile Lys Leu Lys Asn Leu Asn Asn Tyr Ile Ser Leu Phe 180 185 190 Arg Lys Lys Thr Arg Thr Glu Lys Glu Asn Lys Glu Leu Glu Asn Leu 195 200 205 Glu Ile Asn Leu Arg Lys Glu Ile Ala Lys Ala Phe Lys Gly Asn Glu 210 215 220 Gly Tyr Lys Ser Leu Phe Lys Lys Asp Ile Ile Glu Thr Ile Leu Pro 225 230 235 240 Glu Phe Leu Asp Asp Lys Asp Glu Ile Ala Leu Val Asn Ser Phe Asn 245 250 255 Gly Phe Thr Thr Ala Phe Thr Gly Phe Phe Asp Asn Arg Glu Asn Met 260 265 270 Phe Ser Glu Glu Ala Lys Ser Thr Ser Ile Ala Phe Arg Cys Ile Asn 275 280 285 Glu Asn Leu Thr Arg Tyr Ile Ser Asn Met Asp Ile Phe Glu Lys Val 290 295 300 Asp Ala Ile Phe Asp Lys His Glu Val Gln Glu Ile Lys Glu Lys Ile 305 310 315 320 Leu Asn Ser Asp Tyr Asp Val Glu Asp Phe Phe Glu Gly Glu Phe Phe 325 330 335 Asn Phe Val Leu Thr Gln Glu Gly Ile Asp Val Tyr Asn Ala Ile Ile 340 345 350 Gly Gly Phe Val Thr Glu Ser Gly Glu Lys Ile Lys Gly Leu Asn Glu 355 360 365 Tyr Ile Asn Leu Tyr Asn Gln Lys Thr Lys Gln Lys Leu Pro Lys Phe 370 375 380 Lys Pro Leu Tyr Lys Gln Val Leu Ser Asp Arg Glu Ser Leu Ser Phe 385 390 395 400 Tyr Gly Glu Gly Tyr Thr Ser Asp Glu Glu Val Leu Glu Val Phe Arg 405 410 415 Asn Thr Leu Asn Lys Asn Ser Glu Ile Phe Ser Ser Ile Lys Lys Leu 420 425 430 Glu Lys Leu Phe Lys Asn Phe Asp Glu Tyr Ser Ser Ala Gly Ile Phe 435 440 445 Val Lys Asn Gly Pro Ala Ile Ser Thr Ile Ser Lys Asp Ile Phe Gly 450 455 460 Glu Trp Asn Val Ile Arg Asp Lys Trp Asn Ala Glu Tyr Asp Asp Ile 465 470 475 480 His Leu Lys Lys Lys Ala Val Val Thr Glu Lys Tyr Glu Asp Asp Arg 485 490 495 Arg Lys Ser Phe Lys Lys Ile Gly Ser Phe Ser Leu Glu Gln Leu Gln 500 505 510 Glu Tyr Ala Asp Ala Asp Leu Ser Val Val Glu Lys Leu Lys Glu Ile 515 520 525 Ile Ile Gln Lys Val Asp Glu Ile Tyr Lys Val Tyr Gly Ser Ser Glu 530 535 540 Lys Leu Phe Asp Ala Asp Phe Val Leu Glu Lys Ser Leu Lys Lys Asn 545 550 555 560 Asp Ala Val Val Ala Ile Met Lys Asp Leu Leu Asp Ser Val Lys Ser 565 570 575 Phe Glu Asn Tyr Ile Lys Ala Phe Phe Gly Glu Gly Lys Glu Thr Asn 580 585 590 Arg Asp Glu Ser Phe Tyr Gly Asp Phe Val Leu Ala Tyr Asp Ile Leu 595 600 605 Leu Lys Val Asp His Ile Tyr Asp Ala Ile Arg Asn Tyr Val Thr Gln 610 615 620 Lys Pro Tyr Ser Lys Asp Lys Phe Lys Leu Tyr Phe Gln Asn Pro Gln 625 630 635 640 Phe Met Gly Gly Trp Asp Lys Asp Lys Glu Thr Asp Tyr Arg Ala Thr 645 650 655 Ile Leu Arg Tyr Gly Ser Lys Tyr Tyr Leu Ala Ile Met Asp Lys Lys 660 665 670 Tyr Ala Lys Cys Leu Gln Lys Ile Asp Lys Asp Asp Val Asn Gly Asn 675 680 685 Tyr Glu Lys Ile Asn Tyr Lys Leu Leu Pro Gly Pro Asn Lys Met Leu 690 695 700 Pro Lys Val Phe Phe Ser Lys Lys Trp Met Ala Tyr Tyr Asn Pro Ser 705 710 715 720 Glu Asp Ile Gln Lys Ile Tyr Lys Asn Gly Thr Phe Lys Lys Gly Asp 725 730 735 Met Phe Asn Leu Asn Asp Cys His Lys Leu Ile Asp Phe Phe Lys Asp 740 745 750 Ser Ile Ser Arg Tyr Pro Lys Trp Ser Asn Ala Tyr Asp Phe Asn Phe 755 760 765 Ser Glu Thr Glu Lys Tyr Lys Asp Ile Ala Gly Phe Tyr Arg Glu Val 770 775 780 Glu Glu Gln Gly Tyr Lys Val Ser Phe Glu Ser Ala Ser Lys Lys Glu 785 790 795 800 Val Asp Lys Leu Val Glu Glu Gly Lys Leu Tyr Met Phe Gln Ile Tyr 805 810 815 Asn Lys Asp Phe Ser Asp Lys Ser His Gly Thr Pro Asn Leu His Thr 820 825 830 Met Tyr Phe Lys Leu Leu Phe Asp Glu Asn Asn His Gly Gln Ile Arg 835 840 845 Leu Ser Gly Gly Ala Glu Leu Phe Met Arg Arg Ala Ser Leu Lys Lys 850 855 860 Glu Glu Leu Val Val His Pro Ala Asn Ser Pro Ile Ala Asn Lys Asn 865 870 875 880 Pro Asp Asn Pro Lys Lys Thr Thr Thr Leu Ser Tyr Asp Val Tyr Lys 885 890 895 Asp Lys Arg Phe Ser Glu Asp Gln Tyr Glu Leu His Ile Pro Ile Ala 900 905 910 Ile Asn Lys Cys Pro Lys Asn Ile Phe Lys Ile Asn Thr Glu Val Arg 915 920 925 Val Leu Leu Lys His Asp Asp Asn Pro Tyr Val Ile Gly Ile Ala Arg 930 935 940 Gly Glu Arg Asn Leu Leu Tyr Ile Val Val Val Asp Gly Lys Gly Asn 945 950 955 960 Ile Val Glu Gln Tyr Ser Leu Asn Glu Ile Ile Asn Asn Phe Asn Gly 965 970 975 Ile Arg Ile Lys Thr Asp Tyr His Ser Leu Leu Asp Lys Lys Glu Lys 980 985 990 Glu Arg Phe Glu Ala Arg Gln Asn Trp Thr Ser Ile Glu Asn Ile Lys 995 1000 1005 Glu Leu Lys Ala Gly Tyr Ile Ser Gln Val Val His Lys Ile Cys 1010 1015 1020 Glu Leu Val Glu Lys Tyr Asp Ala Val Ile Ala Leu Glu Asp Leu 1025 1030 1035 Asn Ser Gly Phe Lys Asn Ser Arg Val Lys Val Glu Lys Gln Val 1040 1045 1050 Tyr Gln Lys Phe Glu Lys Met Leu Ile Asp Lys Leu Asn Tyr Met 1055 1060 1065 Val Asp Lys Lys Ser Asn Pro Cys Ala Thr Gly Gly Ala Leu Lys 1070 1075 1080 Gly Tyr Gln Ile Thr Asn Lys Phe Glu Ser Phe Lys Ser Met Ser 1085 1090 1095 Thr Gln Asn Gly Phe Ile Phe Tyr Ile Pro Ala Trp Leu Thr Ser 1100 1105 1110 Lys Ile Asp Pro Ser Thr Gly Phe Val Asn Leu Leu Lys Thr Lys 1115 1120 1125 Tyr Thr Ser Ile Ala Asp Ser Lys Lys Phe Ile Ser Ser Phe Asp 1130 1135 1140 Arg Ile Met Tyr Val Pro Glu Glu Asp Leu Phe Glu Phe Ala Leu 1145 1150 1155 Asp Tyr Lys Asn Phe Ser Arg Thr Asp Ala Asp Tyr Ile Lys Lys 1160 1165 1170 Trp Lys Leu Tyr Ser Tyr Gly Asn Arg Ile Arg Ile Phe Arg Asn 1175 1180 1185 Pro Lys Lys Asn Asn Val Phe Asp Trp Glu Glu Val Cys Leu Thr 1190 1195 1200 Ser Ala Tyr Lys Glu Leu Phe Asn Lys Tyr Gly Ile Asn Tyr Gln 1205 1210 1215 Gln Gly Asp Ile Arg Ala Leu Leu Cys Glu Gln Ser Asp Lys Ala 1220 1225 1230 Phe Tyr Ser Ser Phe Met Ala Leu Met Ser Leu Met Leu Gln Met 1235 1240 1245 Arg Asn Ser Ile Thr Gly Arg Thr Asp Val Asp Phe Leu Ile Ser 1250 1255 1260 Pro Val Lys Asn Ser Asp Gly Ile Phe Tyr Asp Ser Arg Asn Tyr 1265 1270 1275 Glu Ala Gln Glu Asn Ala Ile Leu Pro Lys Asn Ala Asp Ala Asn 1280 1285 1290 Gly Ala Tyr Asn Ile Ala Arg Lys Val Leu Trp Ala Ile Gly Gln 1295 1300 1305 Phe Lys Lys Ala Glu Asp Glu Lys Leu Asp Lys Val Lys Ile Ala 1310 1315 1320 Ile Ser Asn Lys Glu Trp Leu Glu Tyr Ala Gln Thr Ser Val Lys 1325 1330 1335 His Gly Gly Ser Lys Gly Pro Pro Pro Ser Pro Pro Pro Pro Gln 1340 1345 1350 Pro Glu Ser Ser Glu Thr Gly Pro Val Ala Val Asp Pro Thr Leu 1355 1360 1365 Arg Arg Arg Ile Glu Pro His Glu Phe Glu Val Phe Phe Asp Pro 1370 1375 1380 Arg Glu Leu Arg Lys Glu Thr Cys Leu Leu Tyr Glu Ile Asn Trp 1385 1390 1395 Gly Gly Arg His Ser Ile Trp Arg His Thr Ser Gln Asn Thr Asn 1400 1405 1410 Lys His Val Glu Val Asn Phe Ile Glu Lys Phe Thr Thr Glu Arg 1415 1420 1425 Tyr Phe Cys Pro Asn Thr Arg Cys Ser Ile Thr Trp Phe Leu Ser 1430 1435 1440 Trp Ser Pro Cys Gly Glu Cys Ser Arg Ala Ile Thr Glu Phe Leu 1445 1450 1455 Ser Arg Tyr Pro His Val Thr Leu Phe Ile Tyr Ile Ala Arg Leu 1460 1465 1470 Tyr His His Ala Asp Pro Arg Asn Arg Gln Gly Leu Arg Asp Leu 1475 1480 1485 Ile Ser Ser Gly Val Thr Ile Gln Ile Met Thr Glu Gln Glu Ser 1490 1495 1500 Gly Tyr Cys Trp Arg Asn Phe Val Asn Tyr Ser Pro Ser Asn Glu 1505 1510 1515 Ala His Trp Pro Arg Tyr Pro His Leu Trp Val Arg Leu Tyr Val 1520 1525 1530 Leu Glu Leu Tyr Cys Ile Ile Leu Gly Leu Pro Pro Cys Leu Asn 1535 1540 1545 Ile Leu Arg Arg Lys Gln Pro Gln Leu Thr Phe Phe Thr Ile Ala 1550 1555 1560 Leu Gln Ser Cys His Tyr Gln Arg Leu Pro Pro His Ile Leu Trp 1565 1570 1575 Ala Thr Gly Leu Lys Ser Gly Gly Ser Lys Arg Thr Ala Asp Gly 1580 1585 1590 Ser Glu Phe 1595 <210> 53 <211> 1596 <212> PRT <213> Artificial <220> <223> Base editor fusion protein <400> 53 Met Lys Arg Thr Ala Asp Gly Ser Glu Phe Glu Ser Pro Lys Lys Lys 1 5 10 15 Arg Lys Val Thr Asn Leu Ser Asp Ile Ile Glu Lys Glu Thr Gly Lys 20 25 30 Gln Leu Val Ile Gln Glu Ser Ile Leu Met Leu Pro Glu Glu Val Glu 35 40 45 Glu Val Ile Gly Asn Lys Pro Glu Ser Asp Ile Leu Val His Thr Ala 50 55 60 Tyr Asp Glu Ser Thr Asp Glu Asn Val Met Leu Leu Thr Ser Asp Ala 65 70 75 80 Pro Glu Tyr Lys Pro Trp Ala Leu Val Ile Gln Asp Ser Asn Gly Glu 85 90 95 Asn Lys Ile Lys Met Leu Ser Gly Gly Ser Gly Gly Ser Gly Gly Ser 100 105 110 Ser Lys Leu Glu Lys Phe Thr Asn Cys Tyr Ser Leu Ser Lys Thr Leu 115 120 125 Arg Phe Lys Ala Ile Pro Val Gly Lys Thr Gln Glu Asn Ile Asp Asn 130 135 140 Lys Arg Leu Leu Val Glu Asp Glu Lys Arg Ala Glu Asp Tyr Lys Gly 145 150 155 160 Val Lys Lys Leu Leu Asp Arg Tyr Tyr Leu Ser Phe Ile Asn Asp Val 165 170 175 Leu His Ser Ile Lys Leu Lys Asn Leu Asn Asn Tyr Ile Ser Leu Phe 180 185 190 Arg Lys Lys Thr Arg Thr Glu Lys Glu Asn Lys Glu Leu Glu Asn Leu 195 200 205 Glu Ile Asn Leu Arg Lys Glu Ile Ala Lys Ala Phe Lys Gly Asn Glu 210 215 220 Gly Tyr Lys Ser Leu Phe Lys Lys Asp Ile Ile Glu Thr Ile Leu Pro 225 230 235 240 Glu Phe Leu Asp Asp Lys Asp Glu Ile Ala Leu Val Asn Ser Phe Asn 245 250 255 Gly Phe Thr Thr Ala Phe Thr Gly Phe Phe Asp Asn Arg Glu Asn Met 260 265 270 Phe Ser Glu Glu Ala Lys Ser Thr Ser Ile Ala Phe Arg Cys Ile Asn 275 280 285 Glu Asn Leu Thr Arg Tyr Ile Ser Asn Met Asp Ile Phe Glu Lys Val 290 295 300 Asp Ala Ile Phe Asp Lys His Glu Val Gln Glu Ile Lys Glu Lys Ile 305 310 315 320 Leu Asn Ser Asp Tyr Asp Val Glu Asp Phe Phe Glu Gly Glu Phe Phe 325 330 335 Asn Phe Val Leu Thr Gln Glu Gly Ile Asp Val Tyr Asn Ala Ile Ile 340 345 350 Gly Gly Phe Val Thr Glu Ser Gly Glu Lys Ile Lys Gly Leu Asn Glu 355 360 365 Tyr Ile Asn Leu Tyr Asn Gln Lys Thr Lys Gln Lys Leu Pro Lys Phe 370 375 380 Lys Pro Leu Tyr Lys Gln Val Leu Ser Asp Arg Glu Ser Leu Ser Phe 385 390 395 400 Tyr Gly Glu Gly Tyr Thr Ser Asp Glu Glu Val Leu Glu Val Phe Arg 405 410 415 Asn Thr Leu Asn Lys Asn Ser Glu Ile Phe Ser Ser Ile Lys Lys Leu 420 425 430 Glu Lys Leu Phe Lys Asn Phe Asp Glu Tyr Ser Ser Ala Gly Ile Phe 435 440 445 Val Lys Asn Gly Pro Ala Ile Ser Thr Ile Ser Lys Asp Ile Phe Gly 450 455 460 Glu Trp Asn Val Ile Arg Asp Lys Trp Asn Ala Glu Tyr Asp Asp Ile 465 470 475 480 His Leu Lys Lys Lys Ala Val Val Thr Glu Lys Tyr Glu Asp Asp Arg 485 490 495 Arg Lys Ser Phe Lys Lys Ile Gly Ser Phe Ser Leu Glu Gln Leu Gln 500 505 510 Glu Tyr Ala Asp Ala Asp Leu Ser Val Val Glu Lys Leu Lys Glu Ile 515 520 525 Ile Ile Gln Lys Val Asp Glu Ile Tyr Lys Val Tyr Gly Ser Ser Glu 530 535 540 Lys Leu Phe Asp Ala Asp Phe Val Leu Glu Lys Ser Leu Lys Lys Asn 545 550 555 560 Asp Ala Val Val Ala Ile Met Lys Asp Leu Leu Asp Ser Val Lys Ser 565 570 575 Phe Glu Asn Tyr Ile Lys Ala Phe Phe Gly Glu Gly Lys Glu Thr Asn 580 585 590 Arg Asp Glu Ser Phe Tyr Gly Asp Phe Val Leu Ala Tyr Asp Ile Leu 595 600 605 Leu Lys Val Asp His Ile Tyr Asp Ala Ile Arg Asn Tyr Val Thr Gln 610 615 620 Lys Pro Tyr Ser Lys Asp Lys Phe Lys Leu Tyr Phe Gln Asn Pro Gln 625 630 635 640 Phe Met Gly Gly Trp Asp Lys Asp Lys Glu Thr Asp Tyr Arg Ala Thr 645 650 655 Ile Leu Arg Tyr Gly Ser Lys Tyr Tyr Leu Ala Ile Met Asp Lys Lys 660 665 670 Tyr Ala Lys Cys Leu Gln Lys Ile Asp Lys Asp Asp Val Asn Gly Asn 675 680 685 Tyr Glu Lys Ile Asn Tyr Lys Leu Leu Pro Gly Pro Asn Lys Met Leu 690 695 700 Pro Lys Val Phe Phe Ser Lys Lys Trp Met Ala Tyr Tyr Asn Pro Ser 705 710 715 720 Glu Asp Ile Gln Lys Ile Tyr Lys Asn Gly Thr Phe Lys Lys Gly Asp 725 730 735 Met Phe Asn Leu Asn Asp Cys His Lys Leu Ile Asp Phe Phe Lys Asp 740 745 750 Ser Ile Ser Arg Tyr Pro Lys Trp Ser Asn Ala Tyr Asp Phe Asn Phe 755 760 765 Ser Glu Thr Glu Lys Tyr Lys Asp Ile Ala Gly Phe Tyr Arg Glu Val 770 775 780 Glu Glu Gln Gly Tyr Lys Val Ser Phe Glu Ser Ala Ser Lys Lys Glu 785 790 795 800 Val Asp Lys Leu Val Glu Glu Gly Lys Leu Tyr Met Phe Gln Ile Tyr 805 810 815 Asn Lys Asp Phe Ser Asp Lys Ser His Gly Thr Pro Asn Leu His Thr 820 825 830 Met Tyr Phe Lys Leu Leu Phe Asp Glu Asn Asn His Gly Gln Ile Arg 835 840 845 Leu Ser Gly Gly Ala Glu Leu Phe Met Arg Arg Ala Ser Leu Lys Lys 850 855 860 Glu Glu Leu Val Val His Pro Ala Asn Ser Pro Ile Ala Asn Lys Asn 865 870 875 880 Pro Asp Asn Pro Lys Lys Thr Thr Thr Leu Ser Tyr Asp Val Tyr Lys 885 890 895 Asp Lys Arg Phe Ser Glu Asp Gln Tyr Glu Leu His Ile Pro Ile Ala 900 905 910 Ile Asn Lys Cys Pro Lys Asn Ile Phe Lys Ile Asn Thr Glu Val Arg 915 920 925 Val Leu Leu Lys His Asp Asp Asn Pro Tyr Val Ile Gly Ile Ala Arg 930 935 940 Gly Glu Arg Asn Leu Leu Tyr Ile Val Val Val Asp Gly Lys Gly Asn 945 950 955 960 Ile Val Glu Gln Tyr Ser Leu Asn Glu Ile Ile Asn Asn Phe Asn Gly 965 970 975 Ile Arg Ile Lys Thr Asp Tyr His Ser Leu Leu Asp Lys Lys Glu Lys 980 985 990 Glu Arg Phe Glu Ala Arg Gln Asn Trp Thr Ser Ile Glu Asn Ile Lys 995 1000 1005 Glu Leu Lys Ala Gly Tyr Ile Ser Gln Val Val His Lys Ile Cys 1010 1015 1020 Glu Leu Val Glu Lys Tyr Asp Ala Val Ile Ala Leu Glu Asp Leu 1025 1030 1035 Asn Ser Gly Phe Lys Asn Ser Arg Val Lys Val Glu Lys Gln Val 1040 1045 1050 Tyr Gln Lys Phe Glu Lys Met Leu Ile Asp Lys Leu Asn Tyr Met 1055 1060 1065 Val Asp Lys Lys Ser Asn Pro Cys Ala Thr Gly Gly Ala Leu Lys 1070 1075 1080 Gly Tyr Gln Ile Thr Asn Lys Phe Glu Ser Phe Lys Ser Met Ser 1085 1090 1095 Thr Gln Asn Gly Phe Ile Phe Tyr Ile Pro Ala Trp Leu Thr Ser 1100 1105 1110 Lys Ile Asp Pro Ser Thr Gly Phe Val Asn Leu Leu Lys Thr Lys 1115 1120 1125 Tyr Thr Ser Ile Ala Asp Ser Lys Lys Phe Ile Ser Ser Phe Asp 1130 1135 1140 Arg Ile Met Tyr Val Pro Glu Glu Asp Leu Phe Glu Phe Ala Leu 1145 1150 1155 Asp Tyr Lys Asn Phe Ser Arg Thr Asp Ala Asp Tyr Ile Lys Lys 1160 1165 1170 Trp Lys Leu Tyr Ser Tyr Gly Asn Arg Ile Arg Ile Phe Arg Asn 1175 1180 1185 Pro Lys Lys Asn Asn Val Phe Asp Trp Glu Glu Val Cys Leu Thr 1190 1195 1200 Ser Ala Tyr Lys Glu Leu Phe Asn Lys Tyr Gly Ile Asn Tyr Gln 1205 1210 1215 Gln Gly Asp Ile Arg Ala Leu Leu Cys Glu Gln Ser Asp Lys Ala 1220 1225 1230 Phe Tyr Ser Ser Phe Met Ala Leu Met Ser Leu Met Leu Gln Met 1235 1240 1245 Arg Asn Ser Ile Thr Gly Arg Thr Asp Val Asp Phe Leu Ile Ser 1250 1255 1260 Pro Val Lys Asn Ser Asp Gly Ile Phe Tyr Asp Ser Arg Asn Tyr 1265 1270 1275 Glu Ala Gln Glu Asn Ala Ile Leu Pro Lys Asn Ala Asp Ala Asn 1280 1285 1290 Gly Ala Tyr Asn Ile Ala Arg Lys Val Leu Trp Ala Ile Gly Gln 1295 1300 1305 Phe Lys Lys Ala Glu Asp Glu Lys Leu Asp Lys Val Lys Ile Ala 1310 1315 1320 Ile Ser Asn Lys Glu Trp Leu Glu Tyr Ala Gln Thr Ser Val Lys 1325 1330 1335 His Ser Glu Ser Pro Thr Asn Gln Gln Pro Glu Pro Gln Trp Thr 1340 1345 1350 Thr Asp Ser Ser Glu Thr Gly Pro Val Ala Val Asp Pro Thr Leu 1355 1360 1365 Arg Arg Arg Ile Glu Pro His Glu Phe Glu Val Phe Phe Asp Pro 1370 1375 1380 Arg Glu Leu Arg Lys Glu Thr Cys Leu Leu Tyr Glu Ile Asn Trp 1385 1390 1395 Gly Gly Arg His Ser Ile Trp Arg His Thr Ser Gln Asn Thr Asn 1400 1405 1410 Lys His Val Glu Val Asn Phe Ile Glu Lys Phe Thr Thr Glu Arg 1415 1420 1425 Tyr Phe Cys Pro Asn Thr Arg Cys Ser Ile Thr Trp Phe Leu Ser 1430 1435 1440 Trp Ser Pro Cys Gly Glu Cys Ser Arg Ala Ile Thr Glu Phe Leu 1445 1450 1455 Ser Arg Tyr Pro His Val Thr Leu Phe Ile Tyr Ile Ala Arg Leu 1460 1465 1470 Tyr His His Ala Asp Pro Arg Asn Arg Gln Gly Leu Arg Asp Leu 1475 1480 1485 Ile Ser Ser Gly Val Thr Ile Gln Ile Met Thr Glu Gln Glu Ser 1490 1495 1500 Gly Tyr Cys Trp Arg Asn Phe Val Asn Tyr Ser Pro Ser Asn Glu 1505 1510 1515 Ala His Trp Pro Arg Tyr Pro His Leu Trp Val Arg Leu Tyr Val 1520 1525 1530 Leu Glu Leu Tyr Cys Ile Ile Leu Gly Leu Pro Pro Cys Leu Asn 1535 1540 1545 Ile Leu Arg Arg Lys Gln Pro Gln Leu Thr Phe Phe Thr Ile Ala 1550 1555 1560 Leu Gln Ser Cys His Tyr Gln Arg Leu Pro Pro His Ile Leu Trp 1565 1570 1575 Ala Thr Gly Leu Lys Ser Gly Gly Ser Lys Arg Thr Ala Asp Gly 1580 1585 1590 Ser Glu Phe 1595 <210> 54 <211> 1596 <212> PRT <213> Artificial <220> <223> Base editor fusion protein <400> 54 Met Lys Arg Thr Ala Asp Gly Ser Glu Phe Glu Ser Pro Lys Lys Lys 1 5 10 15 Arg Lys Val Thr Asn Leu Ser Asp Ile Ile Glu Lys Glu Thr Gly Lys 20 25 30 Gln Leu Val Ile Gln Glu Ser Ile Leu Met Leu Pro Glu Glu Val Glu 35 40 45 Glu Val Ile Gly Asn Lys Pro Glu Ser Asp Ile Leu Val His Thr Ala 50 55 60 Tyr Asp Glu Ser Thr Asp Glu Asn Val Met Leu Leu Thr Ser Asp Ala 65 70 75 80 Pro Glu Tyr Lys Pro Trp Ala Leu Val Ile Gln Asp Ser Asn Gly Glu 85 90 95 Asn Lys Ile Lys Met Leu Ser Gly Gly Ser Gly Gly Ser Gly Gly Ser 100 105 110 Ser Lys Leu Glu Lys Phe Thr Asn Cys Tyr Ser Leu Ser Lys Thr Leu 115 120 125 Arg Phe Lys Ala Ile Pro Val Gly Lys Thr Gln Glu Asn Ile Asp Asn 130 135 140 Lys Arg Leu Leu Val Glu Asp Glu Lys Arg Ala Glu Asp Tyr Lys Gly 145 150 155 160 Val Lys Lys Leu Leu Asp Arg Tyr Tyr Leu Ser Phe Ile Asn Asp Val 165 170 175 Leu His Ser Ile Lys Leu Lys Asn Leu Asn Asn Tyr Ile Ser Leu Phe 180 185 190 Arg Lys Lys Thr Arg Thr Glu Lys Glu Asn Lys Glu Leu Glu Asn Leu 195 200 205 Glu Ile Asn Leu Arg Lys Glu Ile Ala Lys Ala Phe Lys Gly Asn Glu 210 215 220 Gly Tyr Lys Ser Leu Phe Lys Lys Asp Ile Ile Glu Thr Ile Leu Pro 225 230 235 240 Glu Phe Leu Asp Asp Lys Asp Glu Ile Ala Leu Val Asn Ser Phe Asn 245 250 255 Gly Phe Thr Thr Ala Phe Thr Gly Phe Phe Asp Asn Arg Glu Asn Met 260 265 270 Phe Ser Glu Glu Ala Lys Ser Thr Ser Ile Ala Phe Arg Cys Ile Asn 275 280 285 Glu Asn Leu Thr Arg Tyr Ile Ser Asn Met Asp Ile Phe Glu Lys Val 290 295 300 Asp Ala Ile Phe Asp Lys His Glu Val Gln Glu Ile Lys Glu Lys Ile 305 310 315 320 Leu Asn Ser Asp Tyr Asp Val Glu Asp Phe Phe Glu Gly Glu Phe Phe 325 330 335 Asn Phe Val Leu Thr Gln Glu Gly Ile Asp Val Tyr Asn Ala Ile Ile 340 345 350 Gly Gly Phe Val Thr Glu Ser Gly Glu Lys Ile Lys Gly Leu Asn Glu 355 360 365 Tyr Ile Asn Leu Tyr Asn Gln Lys Thr Lys Gln Lys Leu Pro Lys Phe 370 375 380 Lys Pro Leu Tyr Lys Gln Val Leu Ser Asp Arg Glu Ser Leu Ser Phe 385 390 395 400 Tyr Gly Glu Gly Tyr Thr Ser Asp Glu Glu Val Leu Glu Val Phe Arg 405 410 415 Asn Thr Leu Asn Lys Asn Ser Glu Ile Phe Ser Ser Ile Lys Lys Leu 420 425 430 Glu Lys Leu Phe Lys Asn Phe Asp Glu Tyr Ser Ser Ala Gly Ile Phe 435 440 445 Val Lys Asn Gly Pro Ala Ile Ser Thr Ile Ser Lys Asp Ile Phe Gly 450 455 460 Glu Trp Asn Val Ile Arg Asp Lys Trp Asn Ala Glu Tyr Asp Asp Ile 465 470 475 480 His Leu Lys Lys Lys Ala Val Val Thr Glu Lys Tyr Glu Asp Asp Arg 485 490 495 Arg Lys Ser Phe Lys Lys Ile Gly Ser Phe Ser Leu Glu Gln Leu Gln 500 505 510 Glu Tyr Ala Asp Ala Asp Leu Ser Val Val Glu Lys Leu Lys Glu Ile 515 520 525 Ile Ile Gln Lys Val Asp Glu Ile Tyr Lys Val Tyr Gly Ser Ser Glu 530 535 540 Lys Leu Phe Asp Ala Asp Phe Val Leu Glu Lys Ser Leu Lys Lys Asn 545 550 555 560 Asp Ala Val Val Ala Ile Met Lys Asp Leu Leu Asp Ser Val Lys Ser 565 570 575 Phe Glu Asn Tyr Ile Lys Ala Phe Phe Gly Glu Gly Lys Glu Thr Asn 580 585 590 Arg Asp Glu Ser Phe Tyr Gly Asp Phe Val Leu Ala Tyr Asp Ile Leu 595 600 605 Leu Lys Val Asp His Ile Tyr Asp Ala Ile Arg Asn Tyr Val Thr Gln 610 615 620 Lys Pro Tyr Ser Lys Asp Lys Phe Lys Leu Tyr Phe Gln Asn Pro Gln 625 630 635 640 Phe Met Gly Gly Trp Asp Lys Asp Lys Glu Thr Asp Tyr Arg Ala Thr 645 650 655 Ile Leu Arg Tyr Gly Ser Lys Tyr Tyr Leu Ala Ile Met Asp Lys Lys 660 665 670 Tyr Ala Lys Cys Leu Gln Lys Ile Asp Lys Asp Asp Val Asn Gly Asn 675 680 685 Tyr Glu Lys Ile Asn Tyr Lys Leu Leu Pro Gly Pro Asn Lys Met Leu 690 695 700 Pro Lys Val Phe Phe Ser Lys Lys Trp Met Ala Tyr Tyr Asn Pro Ser 705 710 715 720 Glu Asp Ile Gln Lys Ile Tyr Lys Asn Gly Thr Phe Lys Lys Gly Asp 725 730 735 Met Phe Asn Leu Asn Asp Cys His Lys Leu Ile Asp Phe Phe Lys Asp 740 745 750 Ser Ile Ser Arg Tyr Pro Lys Trp Ser Asn Ala Tyr Asp Phe Asn Phe 755 760 765 Ser Glu Thr Glu Lys Tyr Lys Asp Ile Ala Gly Phe Tyr Arg Glu Val 770 775 780 Glu Glu Gln Gly Tyr Lys Val Ser Phe Glu Ser Ala Ser Lys Lys Glu 785 790 795 800 Val Asp Lys Leu Val Glu Glu Gly Lys Leu Tyr Met Phe Gln Ile Tyr 805 810 815 Asn Lys Asp Phe Ser Asp Lys Ser His Gly Thr Pro Asn Leu His Thr 820 825 830 Met Tyr Phe Lys Leu Leu Phe Asp Glu Asn Asn His Gly Gln Ile Arg 835 840 845 Leu Ser Gly Gly Ala Glu Leu Phe Met Arg Arg Ala Ser Leu Lys Lys 850 855 860 Glu Glu Leu Val Val His Pro Ala Asn Ser Pro Ile Ala Asn Lys Asn 865 870 875 880 Pro Asp Asn Pro Lys Lys Thr Thr Thr Leu Ser Tyr Asp Val Tyr Lys 885 890 895 Asp Lys Arg Phe Ser Glu Asp Gln Tyr Glu Leu His Ile Pro Ile Ala 900 905 910 Ile Asn Lys Cys Pro Lys Asn Ile Phe Lys Ile Asn Thr Glu Val Arg 915 920 925 Val Leu Leu Lys His Asp Asp Asn Pro Tyr Val Ile Gly Ile Ala Arg 930 935 940 Gly Glu Arg Asn Leu Leu Tyr Ile Val Val Val Asp Gly Lys Gly Asn 945 950 955 960 Ile Val Glu Gln Tyr Ser Leu Asn Glu Ile Ile Asn Asn Phe Asn Gly 965 970 975 Ile Arg Ile Lys Thr Asp Tyr His Ser Leu Leu Asp Lys Lys Glu Lys 980 985 990 Glu Arg Phe Glu Ala Arg Gln Asn Trp Thr Ser Ile Glu Asn Ile Lys 995 1000 1005 Glu Leu Lys Ala Gly Tyr Ile Ser Gln Val Val His Lys Ile Cys 1010 1015 1020 Glu Leu Val Glu Lys Tyr Asp Ala Val Ile Ala Leu Glu Asp Leu 1025 1030 1035 Asn Ser Gly Phe Lys Asn Ser Arg Val Lys Val Glu Lys Gln Val 1040 1045 1050 Tyr Gln Lys Phe Glu Lys Met Leu Ile Asp Lys Leu Asn Tyr Met 1055 1060 1065 Val Asp Lys Lys Ser Asn Pro Cys Ala Thr Gly Gly Ala Leu Lys 1070 1075 1080 Gly Tyr Gln Ile Thr Asn Lys Phe Glu Ser Phe Lys Ser Met Ser 1085 1090 1095 Thr Gln Asn Gly Phe Ile Phe Tyr Ile Pro Ala Trp Leu Thr Ser 1100 1105 1110 Lys Ile Asp Pro Ser Thr Gly Phe Val Asn Leu Leu Lys Thr Lys 1115 1120 1125 Tyr Thr Ser Ile Ala Asp Ser Lys Lys Phe Ile Ser Ser Phe Asp 1130 1135 1140 Arg Ile Met Tyr Val Pro Glu Glu Asp Leu Phe Glu Phe Ala Leu 1145 1150 1155 Asp Tyr Lys Asn Phe Ser Arg Thr Asp Ala Asp Tyr Ile Lys Lys 1160 1165 1170 Trp Lys Leu Tyr Ser Tyr Gly Asn Arg Ile Arg Ile Phe Arg Asn 1175 1180 1185 Pro Lys Lys Asn Asn Val Phe Asp Trp Glu Glu Val Cys Leu Thr 1190 1195 1200 Ser Ala Tyr Lys Glu Leu Phe Asn Lys Tyr Gly Ile Asn Tyr Gln 1205 1210 1215 Gln Gly Asp Ile Arg Ala Leu Leu Cys Glu Gln Ser Asp Lys Ala 1220 1225 1230 Phe Tyr Ser Ser Phe Met Ala Leu Met Ser Leu Met Leu Gln Met 1235 1240 1245 Arg Asn Ser Ile Thr Gly Arg Thr Asp Val Asp Phe Leu Ile Ser 1250 1255 1260 Pro Val Lys Asn Ser Asp Gly Ile Phe Tyr Asp Ser Arg Asn Tyr 1265 1270 1275 Glu Ala Gln Glu Asn Ala Ile Leu Pro Lys Asn Ala Asp Ala Asn 1280 1285 1290 Gly Ala Tyr Asn Ile Ala Arg Lys Val Leu Trp Ala Ile Gly Gln 1295 1300 1305 Phe Lys Lys Ala Glu Asp Glu Lys Leu Asp Lys Val Lys Ile Ala 1310 1315 1320 Ile Ser Asn Lys Glu Trp Leu Glu Tyr Ala Gln Thr Ser Val Lys 1325 1330 1335 His Ser Glu Ser Pro Ser Lys Gln Gln Pro Glu Pro Lys Ser Ser 1340 1345 1350 Lys Gly Ser Ser Glu Thr Gly Pro Val Ala Val Asp Pro Thr Leu 1355 1360 1365 Arg Arg Arg Ile Glu Pro His Glu Phe Glu Val Phe Phe Asp Pro 1370 1375 1380 Arg Glu Leu Arg Lys Glu Thr Cys Leu Leu Tyr Glu Ile Asn Trp 1385 1390 1395 Gly Gly Arg His Ser Ile Trp Arg His Thr Ser Gln Asn Thr Asn 1400 1405 1410 Lys His Val Glu Val Asn Phe Ile Glu Lys Phe Thr Thr Glu Arg 1415 1420 1425 Tyr Phe Cys Pro Asn Thr Arg Cys Ser Ile Thr Trp Phe Leu Ser 1430 1435 1440 Trp Ser Pro Cys Gly Glu Cys Ser Arg Ala Ile Thr Glu Phe Leu 1445 1450 1455 Ser Arg Tyr Pro His Val Thr Leu Phe Ile Tyr Ile Ala Arg Leu 1460 1465 1470 Tyr His His Ala Asp Pro Arg Asn Arg Gln Gly Leu Arg Asp Leu 1475 1480 1485 Ile Ser Ser Gly Val Thr Ile Gln Ile Met Thr Glu Gln Glu Ser 1490 1495 1500 Gly Tyr Cys Trp Arg Asn Phe Val Asn Tyr Ser Pro Ser Asn Glu 1505 1510 1515 Ala His Trp Pro Arg Tyr Pro His Leu Trp Val Arg Leu Tyr Val 1520 1525 1530 Leu Glu Leu Tyr Cys Ile Ile Leu Gly Leu Pro Pro Cys Leu Asn 1535 1540 1545 Ile Leu Arg Arg Lys Gln Pro Gln Leu Thr Phe Phe Thr Ile Ala 1550 1555 1560 Leu Gln Ser Cys His Tyr Gln Arg Leu Pro Pro His Ile Leu Trp 1565 1570 1575 Ala Thr Gly Leu Lys Ser Gly Gly Ser Lys Arg Thr Ala Asp Gly 1580 1585 1590 Ser Glu Phe 1595 <210> 55 <211> 1598 <212> PRT <213> Artificial <220> <223> Base editor fusion protein <400> 55 Met Lys Arg Thr Ala Asp Gly Ser Glu Phe Glu Ser Pro Lys Lys Lys 1 5 10 15 Arg Lys Val Thr Asn Leu Ser Asp Ile Ile Glu Lys Glu Thr Gly Lys 20 25 30 Gln Leu Val Ile Gln Glu Ser Ile Leu Met Leu Pro Glu Glu Val Glu 35 40 45 Glu Val Ile Gly Asn Lys Pro Glu Ser Asp Ile Leu Val His Thr Ala 50 55 60 Tyr Asp Glu Ser Thr Asp Glu Asn Val Met Leu Leu Thr Ser Asp Ala 65 70 75 80 Pro Glu Tyr Lys Pro Trp Ala Leu Val Ile Gln Asp Ser Asn Gly Glu 85 90 95 Asn Lys Ile Lys Met Leu Ser His Pro Pro Gln Glu Pro Pro Gln Ser 100 105 110 Asn Leu Ser Lys Leu Glu Lys Phe Thr Asn Cys Tyr Ser Leu Ser Lys 115 120 125 Thr Leu Arg Phe Lys Ala Ile Pro Val Gly Lys Thr Gln Glu Asn Ile 130 135 140 Asp Asn Lys Arg Leu Leu Val Glu Asp Glu Lys Arg Ala Glu Asp Tyr 145 150 155 160 Lys Gly Val Lys Lys Leu Leu Asp Arg Tyr Tyr Leu Ser Phe Ile Asn 165 170 175 Asp Val Leu His Ser Ile Lys Leu Lys Asn Leu Asn Asn Tyr Ile Ser 180 185 190 Leu Phe Arg Lys Lys Thr Arg Thr Glu Lys Glu Asn Lys Glu Leu Glu 195 200 205 Asn Leu Glu Ile Asn Leu Arg Lys Glu Ile Ala Lys Ala Phe Lys Gly 210 215 220 Asn Glu Gly Tyr Lys Ser Leu Phe Lys Lys Asp Ile Ile Glu Thr Ile 225 230 235 240 Leu Pro Glu Phe Leu Asp Asp Lys Asp Glu Ile Ala Leu Val Asn Ser 245 250 255 Phe Asn Gly Phe Thr Thr Ala Phe Thr Gly Phe Phe Asp Asn Arg Glu 260 265 270 Asn Met Phe Ser Glu Glu Ala Lys Ser Thr Ser Ile Ala Phe Arg Cys 275 280 285 Ile Asn Glu Asn Leu Thr Arg Tyr Ile Ser Asn Met Asp Ile Phe Glu 290 295 300 Lys Val Asp Ala Ile Phe Asp Lys His Glu Val Gln Glu Ile Lys Glu 305 310 315 320 Lys Ile ...

Claims

1. A polypeptide consisting of the amino acid sequence of SEQ ID NO:

1.

2. A fusion protein comprising a Cas12a domain, a polypeptide of interest, and the amino acid sequence of SEQ ID NO: 1, wherein the structure of the fusion protein is as follows: apolipoprotein B mRNA editing complex (APOBEC) domain - SEQ ID NO: 1 - Cas12a domain - GS linker - uracil-DNA glycosylase inhibitor (UGI), from N-terminus to C-terminus.

3. The fusion protein of claim 2, wherein the APOBEC domain is a rat or human APOBEC domain.

4. The fusion protein of claim 3, wherein the APOBEC domain has the amino acid sequence of SEQ ID NO:

46.

5. The fusion protein of claim 3, wherein the APOBEC domain has the amino acid sequence of SEQ ID NO:

47.

6. The fusion protein of claim 2, wherein the GS linker is (GSS)n, S(GGS)n, SGGS, SGGSGGSGGS, SGSETPGTSESATPES, or SGGSSGGSSGSETPGTSESATPESSGGSSGGS.

7. A polynucleotide encoding the fusion protein of any one of claims 2 to 6.

8. The polynucleotide of claim 7, wherein the polynucleotide is codon-optimized for expression in a biological organism.

9. The polynucleotide of claim 8, wherein the biological organism is an animal, a plant, a fungus, an archaeon, or a bacterium.

10. A complex comprising the fusion protein of any one of claims 2 to 6 and a guide nucleic acid.

11. A nucleic acid construct encoding the complex of claim 10.

12. A composition comprising the fusion protein of any one of claims 2 to 6 and a guide nucleic acid.

13. An expression cassette or vector comprising the polynucleotide of any one of claims 7 to 9 or the nucleic acid construct of claim 11.

14. A type V clustered regularly interspaced short palindromic repeat (CRISPR-Cas) system comprising: (a) A fusion protein comprising a Cas12a domain, a linker consisting of the amino acid sequence of SEQ ID NO: 1, and a polypeptide of interest, wherein the Cas12a domain is linked to the polypeptide of interest via the amino acid sequence of SEQ ID NO: 1, and wherein the fusion protein is the fusion protein of any one of claims 2 to 6; or a nucleic acid encoding the fusion protein; and (b) A guide nucleic acid comprising a spacer sequence and a repeat sequence, wherein the guide nucleic acid is capable of forming a complex with the Cas12a domain of the fusion protein, and the spacer sequence is capable of hybridizing to a target nucleic acid, thereby guiding the Cas12a domain and the polypeptide of interest to the target nucleic acid, whereby the system is capable of modifying or regulating the target nucleic acid.

15. The system of claim 14, wherein (a) and (b) are comprised in one or more expression cassettes and / or vectors.

16. A cell comprising the polynucleotide of any one of claims 7 to 9, the nucleic acid construct of claim 11, the expression cassette or vector of claim 13, or the system of any one of claims 14 to 15, wherein the cell is neither a plant variety nor an animal variety.

17. A method of modifying a target nucleic acid, comprising contacting the target nucleic acid with: (a) (i) a fusion protein of any one of claims 2 to 6, and (a) (ii) a guide nucleic acid; (b) the complex of claim 10, and a guide nucleic acid; (c) a composition comprising a fusion protein of any one of claims 2 to 6 and a guide nucleic acid; and / or (d) the system of claim 14, thereby modifying the target nucleic acid, wherein the method is not a method of treating a disease.

18. A method of modifying a target nucleic acid, comprising contacting a cell or cell-free system comprising the target nucleic acid with: (a) (i) a polynucleotide encoding a fusion protein of any one of claims 2 to 6, or an expression cassette or vector comprising the same, and (a) (ii) a guide nucleic acid, or an expression cassette or vector comprising the same; and / or (b) a nucleic acid construct encoding the complex of claim 10, or an expression cassette or vector comprising the same, thereby modifying the target nucleic acid, wherein the conditions are such that the fusion protein is expressed and forms a complex with the guide nucleic acid, and the complex hybridizes with the target nucleic acid, wherein the method is not a method of treating a disease.

19. A method of editing a target nucleic acid, comprising contacting the target nucleic acid with: (a) (i) a fusion protein of any one of claims 2 to 6, and (a) (ii) a guide nucleic acid; (b) the complex of claim 10; (c) a composition comprising a fusion protein of any one of claims 2 to 6 and a guide nucleic acid; and / or (d) the system of claim 14, wherein the cytosine deaminase domain converts cytosine (C) in the target nucleic acid to thymine (T), thereby editing the target nucleic acid to generate a mutation, wherein the method is not a method of treating a disease.

20. A method of editing a target nucleic acid, comprising contacting a cell or cell-free system comprising the target nucleic acid with: (a) (i) a polynucleotide encoding a fusion protein of any one of claims 2 to 6, or an expression cassette or vector comprising the same, and (a) (ii) a guide nucleic acid, or an expression cassette or vector comprising the same; (b) a nucleic acid construct encoding the complex of claim 10, or an expression cassette or vector comprising the same; and / or (c) the system of claim 15, wherein the conditions are such that the fusion protein is expressed and forms a complex with the guide nucleic acid, and the complex hybridizes with the target nucleic acid, wherein the cytosine deaminase domain converts cytosine (C) in the target nucleic acid to thymine (T), thereby editing the target nucleic acid, wherein the method is not a method of treating a disease.

21. The method of claim 19 or 20, wherein the point mutation is a C→T transition in the sense strand of the target nucleic acid or a G→A transition in the antisense strand of the target nucleic acid.

22. A kit comprising the fusion protein of any one of claims 2 to 6.

23. The kit of claim 22, further comprising instructions for its use.

24. A kit comprising the polynucleotide of any one of claims 7 to 9 and / or an expression cassette or vector comprising the same.

25. The kit of claim 24, further comprising instructions for its use.

26. The kit of any one of claims 22 to 25, further comprising a Cas12a guide nucleic acid and / or an expression cassette or vector comprising the same.

27. The kit of claim 26, wherein the guide nucleic acid comprises a cloning site for cloning a nucleic acid sequence identical or complementary to the target nucleic acid sequence into the backbone of the guide nucleic acid.

28. The kit of any one of claims 22 to 23, wherein the fusion protein further comprises one or more nuclear localization signals.

29. The kit of any one of claims 24 to 25, wherein the polynucleotide further encodes one or more selectable markers.

30. The kit of any one of claims 24 to 25, wherein the polynucleotide is mRNA and encodes one or more introns within the encoded fusion protein.

Images