Bacteria with modified prophage

Abstract

Disclosed herein are methods and compositions for increasing culture stability of a bacterial culture comprising a plurality of a modified bacterium comprising a modified prophage. Also disclosed herein are methods and compositions for decreasing the likelihood of replication of a modified prophage in a bacterial genome as compared to the likelihood of replication of a corresponding prophage lacking the prophage modification in a corresponding bacterial genome. Also disclosed herein are methods and compositions for preventing excision of a prophage in a bacterium. Also disclosed herein are methods and compositions for generating a modified prophage having an impaired ability to lyse a host bacterial cell.

Description

[0001] Attorney Docket No. PIV-00053 WO

[0002] Bacteria with Modified Prophage

[0003] CROSS-REFERENCE TO A RELATED APPLICATION This application claims priority to U. S. Provisional Patent Application No.

[0004] 63 / 734,626, filed December 16, 2024, the entire contents of which are herein incorporated by reference.

[0005] INCORPORATION OF THE SEQUENCE LISTING

[0006] The contents of the electronic sequence listing (PIV-00053 WO SeqList ST26.xml, Size: 188,015 bytes; and Date of Creation December 8, 2025) are herein incorporated by reference in their entirety.

[0007] TECHNICAL FIELD

[0008] Bacterial strains that have been gene-edited for improved culturing stability, and methods for their use are described herein. Such bacterial strains, and compositions containing the bacterial strains, can be used to provide nitrogen to plants.

[0009] BACKGROUND

[0010] Bacteriophages are viruses that can infect bacterial cells. The bacteriophage genome can be integrated in the bacterial genome as a prophage until a trigger occurs that induces the prophage to excise and replicate, thereby producing bacteriophage that can cause cell death and infect neighboring cells.

[0011] Bacterial strains can be useful for production of many types of valuable chemical and biological compounds. However, some bacterial strains may contain prophages that can lay dormant but then can be activated by various triggers, causing bacterial production cultures to die off. This can increase the costs and complexity of commercial production of products using such bacterial strains.

[0012] In recent years, DNA technology has been developed that can be useful in determining when a bacterial culture is negatively impacted by the activation of a prophage. Current methods of limiting bacteriophage activation, including prophage activation, in bacterial strains include adapted factory design, improved sanitation, process changes, the use of specific culture media, bacterial strain rotation, and the use of phage-resistant strains (e.g., strains generated using CRISPR / Cas systems, abortive infection systems, and other bacteriophage defense systems). However, there remains a need for reliable methods of Attorney Docket No. PIV-00053 WO

[0013] deactivating or preventing the activation of prophages to produce bacterial strains that can be grown without the threat of bacterial culture loss due to prophage activation.

[0014] SUMMARY

[0015] This document provides methods and materials for inactivating or preventing activation of prophages to produce bacterial strains that can be grown (e.g., during commercial or industrial production) without the threat of loss of the bacterial cultures due to prophage activation. The methods provided herein can be used, for example, to produce strains of bacteria that are less likely to activate prophage during bacterial fermentation. The resulting fermentation cultures therefore are less likely to die, which would otherwise cause significant loss of time and materials to companies or researchers performing the bacterial fermentations, and could result in product variability. The methods provided herein can limit prophage activation by, for example, grounding the prophage, or limiting / removing the prophage's ability to excise out of a bacterial genome. Prophage containing specific genetic modifications that result in their grounding in the bacterial host genome also are described herein.

[0016] Disclosed herein are methods of increasing culture stability of a bacterial culture including a plurality of a modified bacterium. In some cases, the method includes a) identifying a bacterium including a genome having a prophage; and b) modifying an attL DNA sequence of the prophage, an attR DNA sequence of the prophage, and / or a nucleic acid sequence encoding a recombinase polypeptide within the prophage, thereby producing a bacterium having a bacterial genome comprising a modified prophage, where the modified prophage includes from about 51% to about 99.99% of the nucleotide sequence corresponding to the prophage lacking the modification, and where the modified prophage cannot excise from the bacterial genome, and where the culture stability of the bacterial culture including the plurality of the modified bacterium is increased in comparison to a level of culture stability in a corresponding bacterial culture including a plurality of the bacterium including the prophage lacking the modification. In some cases, the culture stability is measured by a decrease in prophage activation after treatment with prophage activating conditions.

[0017] Also disclosed herein are methods of decreasing the likelihood of replication of a modified prophage in a bacterial genome as compared to the likelihood of replication of a corresponding prophage lacking the modification in a corresponding bacterial genome. In Attorney Docket No. PIV-00053 WO

[0018] some cases, the method includes a) identifying a bacterium including a genome having the prophage; b) modifying an attL DNA sequence of the prophage, an attR DNA sequence of the prophage, and / or a nucleic acid sequence encoding a recombinase polypeptide within the prophage, thereby producing a bacterium having a bacterial genome comprising a modified prophage, where the modified prophage includes from about 51% to about 99.99% of the nucleotide sequence corresponding to the prophage lacking the modification, and where the modified prophage cannot excise from the bacterial genome; and c) generating a bacterial culture including a plurality of the bacterium having the modified prophage, where the plurality of the bacterium having the modified prophage has a decreased likelihood of replication of the prophage compared to a level of the likelihood of replication in a corresponding bacterial culture including a plurality of the bacterium including the prophage lacking the modification. In some cases, the likelihood of replication is measured by whole genome sequencing after treatment with prophage activating conditions.

[0019] Also disclosed herein are methods of preventing excision of a prophage in a bacterium. In some cases, the method includes modifying an attL DNA sequence of the prophage, an attR DNA sequence of the prophage, and / or a nucleic acid sequence encoding a recombinase polypeptide within the prophage, thereby producing a bacterium having a bacterial genome comprising a modified prophage, where the modified prophage includes from about 51% to about 99.99% of the nucleotide sequence corresponding to the prophage lacking the modification, and where the modified prophage cannot excise from the bacterial genome.

[0020] Any of the methods disclosed herein can also include growing a bacterial culture in a condition that activates the prophage, where the bacterial culture includes a plurality of bacteria having a bacterial genome with a modified prophage; and determining a lack of prophage replication. In some cases, the condition that activates the prophage comprises mitomycin C or late stationary phase growth.

[0021] Also disclosed herein are methods of generating a modified prophage having an impaired ability to lyse a host bacterial cell. In some cases, the method includes modifying an attL DNA sequence of a prophage within a bacterial genome, an attR DNA sequence of the prophage within the bacterial genome, and / or a nucleic acid sequence encoding a recombinase polypeptide of the prophage within the bacterial genome, thereby producing a bacterium having a bacterial genome comprising the modified prophage, where the modified prophage includes from about 51% to about 99.99% of the nucleotide sequence Attorney Docket No. PIV-00053 WO

[0022] corresponding to the prophage lacking the modification, and where the modified prophage cannot excise from the bacterial genome.

[0023] Also disclosed herein are genetically engineered bacteria that include a modified prophage, where the modified prophage includes at least one modification selected from a modified attL DNA sequence, a modified attR DNA sequence, a modified nucleic acid sequence encoding a recombinase polypeptide, a partially deleted attL DNA sequence, a partially deleted attR DNA sequence, a partially deleted nucleic acid sequence encoding the recombinase polypeptide, a full deletion of an attL DNA sequence, a full deletion of an attR DNA sequence, and / or a full deletion of a nucleic acid sequence encoding the recombinase polypeptide; where the modified prophage includes from about 51% to about 99.99% of a nucleotide sequence of the prophage; and where the modified prophage cannot excise from the genome of the genetically engineered bacterium

[0024] Also disclosed herein are modified prophage having a DNA sequence that is at least 90% identical to SEQ ID NO: 9-11 and cannot be excised from a host bacterial cell. In some cases, all of the phage recombinase sequences that are present in the bacterial genome are inactivated or removed. In any of the methods or compositions disclosed herein, the nucleic acid sequence encoding the recombinase polypeptide, the attL DNA sequence, and the attR DNA sequence are completely deleted. In any of the methods or compositions disclosed herein, the nucleic acid sequence encoding the recombinase polypeptide, the attL DNA sequence, and the attR DNA sequence are partially deleted. In any of the methods or compositions disclosed herein, the nucleic acid sequence encoding the recombinase polypeptide is completely deleted, and where the attL DNA sequence and the attR DNA sequence are partially deleted. In any of the methods or compositions disclosed herein, the nucleic acid sequence encoding the recombinase polypeptide and the attL DNA sequence are completely deleted, and where the attR DNA sequence is partially deleted. In any of the methods or compositions disclosed herein, the nucleic acid sequence encoding the recombinase polypeptide and the attR DNA sequence are completely deleted, and where the attL DNA sequence is partially deleted. In any of the methods or compositions disclosed herein, the nucleic acid sequence encoding the recombinase polypeptide is partially deleted, and where the attL DNA sequence and the attR DNA sequence are completely deleted. In any of the methods or compositions disclosed herein, the nucleic acid sequence encoding the recombinase polypeptide and the attL DNA sequence are partially deleted, and where the attR DNA sequence is completely deleted. In any of the methods or compositions Attorney Docket No. PIV-00053 WO

[0025] disclosed herein, the nucleic acid sequence encoding the recombinase polypeptide and the attR DNA sequence are partially deleted, and where the attL DNA sequence is completely deleted. In any of the methods or compositions disclosed herein, the recombinase polypeptide encoded by the modified nucleic acid sequence, the modified attL DNA sequence and the modified attR DNA sequence are inactive. In any of the methods or compositions disclosed herein, the nucleic acid sequence encoding the recombinase polypeptide, if present, includes a xerC2 gene or a portion or homolog thereof, a xerC3 gene or a portion or homolog thereof, or a xerD gene or a portion or homolog thereof.

[0026] In any of the methods or compositions disclosed herein, the bacterium is a gram-negative bacterium. In any of the methods or compositions disclosed herein, the bacterium is a gram-positive bacterium. In any of the methods or compositions disclosed herein, the bacterium is a nitrogen-fixing bacterium. In any of the methods or compositions disclosed herein, the nitrogen-fixing bacterium is of a genus selected from Azospirillum, Kosakonia, Klebsiella, and Paraburkholderia. In any of the methods or compositions disclosed herein, the nitrogen-fixing bacterium is of the species Kosakonia oryzendophytica or Kosakonia arachidis. In some cases, the nitrogen-fixing bacterium is of the species Kosakonia oryzendophytica, strain 5812 (ATCC Accession No. PTA-127355). In any of the methods or compositions disclosed herein, the nitrogen-fixing bacterium is of the species Azospirillum palustre. In any of the methods or compositions disclosed herein, the nitrogen-fixing bacterium also includes a modification in one or more genes involved in nitrogen fixation or nitrogen assimilation.

[0027] In any of the methods or compositions disclosed herein, the modified prophage has a DNA sequence that has at least 90% sequence identity to SEQ ID NO: 9-11.

[0028] In any of the methods or compositions disclosed herein, the modified bacterium has an increased competitive fitness when co-cultured with a corresponding bacterial cell lacking the prophage. In any of the methods or compositions disclosed herein, the increased competitive fitness is determined under an environmental stress condition. In any of the methods or compositions disclosed herein, the environmental stress condition is selected from the group consisting of nutrient stress, salt stress, osmotic stress, cold stress, heat stress, excess UV radiation, the presence of antibiotics, and DNA damage.

[0029] In any of the methods or compositions disclosed herein, the modified bacterium has an increased cell growth rate in comparison to a corresponding bacterium lacking the Attorney Docket No. PIV-00053 WO

[0030] prophage. In any of the methods or compositions disclosed herein, the growth rate is measured in a liquid culture medium, in a soil sample, or in a field.

[0031] The use of the terms "a" and "an" and "the" and similar referents in the context of describing the disclosure (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. The terms "comprising," "having," "including," and "containing" are to be construed as open-ended terms (i.e., meaning "including, but not limited to,") unless otherwise noted.

[0032] Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. For example, if the range 10-15 is disclosed, then 11, 12, 13, and 14 are also disclosed.

[0033] All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., "such as") provided herein, is intended merely to better illuminate the disclosure and does not pose a limitation on the scope of the disclosure unless otherwise claimed.

[0034] As used herein, the term "about” is used synonymously with the term "approximately." Illustratively, the use of the term "about" with regard to an amount indicates that values slightly outside the cited values, e.g., plus or minus 0.1% to 10%.

[0035] No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the disclosure.

[0036] 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 disclosure belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the subject matter herein, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting. Attorney Docket No. PIV-00053 WO

[0037] The details of one or more embodiments of the invention are set forth in the accompanying drawings and the description below. Other features, objects, and advantages of the invention will be apparent from the description and drawings, and from the claims.

[0038] All publications, patent applications, patents, sequences, database entries, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control.

[0039] DESCRIPTION OF THE DRAWINGS FIG. 1 is a schematic showing a bacteriophage lifecycle.

[0040] FIG. 2A - FIG. 2C is a series of exemplary schematics showing prophage activation and excision from a host genome.

[0041] FIG. 3 is a schematic showing a general map of a prophage sequence.

[0042] FIGs. 4A-4B show the fitness of bacterial strains with grounded prophage compared to full prophage deletions as determined by flow cytometry (FIG. 4A) or by colony forming unit (CFU) counting (FIG. 4B).

[0043] FIG. 5 is a graph of the amount of ammonium produced (millimolar (mM)) per optical density of bacteria (NH4+ / OD) by ammonium secretion mutants.

[0044] FIGs. 6A-6C demonstrate phage grounding in Kosakonia oryzendophytica strain 5812. FIG. 6A is a graph showing the increased read coverage in two replicates of WT 5812 strain. FIG. 6B is a graph showing that prophage replication levels are inhibited in strain 5812-7767. FIG. 6C is a graph showing that prophage replication levels are inhibited in strain 5812-7663.

[0045] FIG. 7A is a table describing the mutations in Kosakonia arachidis strains 1661-5402 and 1661-11364. FIG. 7B is a graph showing the increased read coverage of K. arachidis strain 1661-5402. FIG. 7C is a graph showing that prophage replication levels in K. arachidis strain 1661-11364.

[0046] FIG. 8 is a graph of the optical density (OD) of the indicated E. coli bacterial strains (background: BG; 1-C6, and 1-C7).

[0047] FIG. 9 is a graph of the growth over time of the indicated E. coli bacterial strains (BG, 1-C6, and 1-C7) in LB broth.

[0048] FIG. 10 is a graph of the growth over time of the indicated E. coli bacterial strains (BG, 1-C6, and 1-C7) in M9 medium with glucose. Attorney Docket No. PIV-00053 WO

[0049] DETAILED DESCRIPTION

[0050] This document provides methods of increasing bacterial culture stability by genetically modifying bacteria. Bacteria having such genetic modifications can be more stable when grown in large scale cultures (e.g., scaled-up cultures), or when applied to soil to improve crop growth.

[0051] Bacteriophages (also called phages) are viruses that infect bacteria and have been estimated to be present in 47-70% of bacteria. Prophages are bacteriophages that can be integrated into the host genome. Bacteriophages capable of this lifestyle are called temperate or lysogenic bacteriophages. A description of a typical phage lifecycle is shown in FIGs. 1A-1E. The phage sequence can remain in the host genome as the host bacterium grows and divides without killing the cell (FIG. 1A). Upon exposure to a stress (indicated in the figure by a lightning bolt), the prophage can excise from the genome (be "triggered"), circularize (FIG. IB), and replicate using the cell's machinery (FIG. 1C). Once progeny phages are assembled (FIG. ID), the phage then lyses, releasing virions (virus particles) that go on to infect other nearby cells (FIG. IE).

[0052] While conditions that trigger prophages to activate and excise from a host genome can vary widely, the molecular mechanism of excision is known (FIG. 2). Briefly, the prophage recombinase protein binds the recombinase directionality factor (RDF) along with the left and right attachment sites (attL and attR), excising the prophage genome from the bacterial host genome, and allowing for replication and assembly of additional bacteriophages (called virions) that can escape a host cell, for example, by lysing and killing the bacterial host cell. When virions infect other host cells, the bacteriophage genome can integrate into the genome at various locations indicated by attachment phage (attP) sites, thus becoming a prophage until triggered again to continue the bacteriophage's life cycle. This process, particularly the reactivation or triggering of prophage and subsequent host cell lysis, can cause the host cell culture to die (or "crash") and can be a significant problem for industries that rely on large-scale bacterial cell culturing or bacterial cell fermentation.

[0053] The presence of activatable prophages can pose risks to companies using bacteria as products or to make components of products (e.g., biologies). These risks include making bacterial strains unpredictable and stymying strain engineering efforts, making experimental outcomes difficult to interpret, killing bacterial cultures during production of biologies, and decreasing shelf-life because the stresses (including changes in pH, waste products, and Attorney Docket No. PIV-00053 WO

[0054] fluctuating temperatures) experienced during storage over time may trigger a prophage resulting in non-viable lysed bacterial cells when a customer uses the product.

[0055] Described herein are methods for selectively modifying a prophage to generate a "grounded" prophage (i.e., a prophage that cannot be excised from a bacterial host genome and therefore cannot be activated). Surprisingly, modified prophages generated by the methods provided herein not only limit the risks to the bacterial culture of the host bacteria containing the grounded prophage, but, in some cases, also can increase the competitive fitness of the modified bacteria. " Phage grounding" refers to the inactivation or inhibition of a prophage by genetically modifying or removing the attachment sites or phage recombinase that allow the prophage to excise from the genome (FIG. 2).

[0056] Modified Prophage

[0057] Disclosed herein are modified prophages containing a genetic modification in the DNA sequence of the prophage. For example, the genetic modification can result in a grounded prophage, a prophage has a decreased likelihood of excising from a host bacterial cell genome compared to a level of likelihood of excising from a corresponding host bacterial cell genome containing a prophage that has not been modified, a prophage that cannot excise from a host bacterial cell genome, or a prophage that has a decreased likelihood of replication in a host bacterial cell compared to a level of likelihood of replicating in a corresponding host bacterial cell containing a prophage that has not been modified. For example, grounding a prophage in a host bacterial cell genome can include genetic modifications to an attL DNA sequence of the prophage, an attR DNA sequence of the prophage, a nucleic acid encoding a phage recombinase polypeptide, and any combinations thereof. In some cases, genetic modifications can include a genetic modification to an attL DNA sequence of the prophage. In some cases, genetic modifications can include a genetic modification to an attR DNA sequence of the prophage. In some cases, genetic modifications can include a genetic modification to a nucleic acid encoding a recombinase polypeptide in the prophage. In some cases, genetic modifications can include a genetic modification to an attL DNA sequence of the prophage and a genetic modification to an attR DNA sequence of the prophage. In some cases, genetic modifications can include a genetic modification to an attL DNA sequence of the prophage and a genetic modification to a nucleic acid encoding a recombinase polypeptide in the prophage. In some cases, genetic modifications can include a genetic modification to an attR DNA sequence of the prophage and a genetic modification Attorney Docket No. PIV-00053 WO

[0058] to a nucleic acid encoding a recombinase polypeptide in the prophage. In some cases, genetic modifications can include a genetic modification to an attL DNA sequence of the prophage, a genetic modification to an attR DNA sequence of the prophage, and a genetic modification to a nucleic acid encoding a recombinase polypeptide in the prophage.

[0059] Genetic modifications can include a complete deletion, a partial deletion, or an inactivation of a DNA sequence or a nucleic acid in the prophage or host bacterial cell genome. A complete deletion includes removing the entire target DNA sequence or nucleic acid from a prophage or host bacterial cell genome. A complete deletion can also include deletion of DNA sequences or nucleic acid 3' or 5' of the target sequence from a prophage or host bacterial cell genome. A partial deletion includes removing part of a target DNA sequence or nucleic acid from a prophage or host bacterial cell genome. A partial deletion can also include deletion of DNA sequences or nucleic acid 3' or 5' of the target sequence from a prophage or host bacterial cell genome. An inactivation of a DNA sequence or a nucleic acid includes modifications (e.g., point mutations, deletion, substitution, insertion) that prevents the target DNA sequence or nucleic acid from performing its function.

[0060] For example, genetic modifications can include a complete deletion, a partial deletion, or an inactivation of an attL DNA sequence of the prophage, an attR DNA sequence of the prophage, a nucleic acid encoding a phage recombinase polypeptide, and any combinations thereof. In some cases, genetic modifications can include a complete deletion, a partial deletion, or an inactivation of an attL DNA sequence of the prophage. In some cases, genetic modifications can include a complete deletion, a partial deletion, or an inactivation of an attR DNA sequence of the prophage. In some cases, genetic modifications can include a complete deletion, a partial deletion, or an inactivation of a nucleic acid encoding a recombinase polypeptide in the prophage. In some cases, genetic modifications can include a complete deletion, a partial deletion, or an inactivation of an attL DNA sequence of the prophage and a complete deletion, a partial deletion, or an inactivation of an attR DNA sequence of the prophage. In some cases, genetic modifications can include a complete deletion, a partial deletion, or an inactivation of an attL DNA sequence of the prophage and a complete deletion, a partial deletion, or an inactivation of to a nucleic acid encoding a recombinase polypeptide in the prophage. In some cases, genetic modifications can include a complete deletion, a partial deletion, or an inactivation of an attR DNA sequence of the prophage and a complete deletion, a partial deletion, or an inactivation of a nucleic acid encoding a recombinase polypeptide in the prophage. In some cases, genetic Attorney Docket No. PIV-00053 WO

[0061] modifications can include a complete deletion, a partial deletion, or an inactivation of an attL DNA sequence of the prophage, a complete deletion, a partial deletion, or an inactivation of an attR DNA sequence of the prophage, and a complete deletion, a partial deletion, or an inactivation of a nucleic acid encoding a recombinase polypeptide in the prophage.

[0062] Any appropriate attL DNA sequence can be modified. Exemplary attL DNA sequences are provided in Table 1.

[0063] Any appropriate attR DNA sequence can be modified. Exemplary attR DNA sequences are provided in Table 1.

[0064] Any appropriate nucleic acid encoding a phage recombinase polypeptide can be modified. Phage recombinase polypeptides can include a XerC2 polypeptide or homolog thereof, a XerC3 polypeptide or homolog thereof, or a XerD polypeptide or homolog thereof. Exemplary nucleic acids encoding a recombinase and recombinase polypeptides are provided in Table 1.

[0065] Table 1. Exemplary DNA sequences and polypeptides.

[0066] SEQ ID Description Sequence from prophage encoded by Kosakonia oryzendophytico strain 5812

[0067] NO:

[0068] 1 attL DNA GGGCGG nCGCGGTCGGGAAACGTTGTGAAAATGTGCACTGCCGTrCACTAAGCAAGCAACCGG sequence CGGAAAAGTCATTGACTCACCAGGCGCTGACCGTATAATTCCAGGCGTTTCCCCTTGCAGGGAGC ATCATCCACGAAATGCGCCCTTAGCTCAGCTGGATAGAGCAACGGCC7TCTAAGCCGTAGGTCAC AGGTTCGAACCCTGTAGGGCGTACCATTTAAAATCAAATGCTTACGCAAGTCTCAATCCAGCCTGA ITTCCTCCtTGTGTCGTATrrGTGTCATGGTrGCCAAAAATGGCATCAATmCCGCGCGTGCTCAC TCAGGTGATTAGGCGCGAGG

[0069] 2 attR DNA GCATAGATGCACTTGAAAGAACCGTAAAAGCATGTGTTGTCATGTAATGACACAAACCCGGCGCG sequence AGCGCGGGAAAAACAGTGATATTACAATGCCATAAATAACGCTCTACGTTCTTCTAAGCCGTAGG TCACAGGTTCGAACCCTGTAGGGCGTACCATTTCCCAGTAAACAGCCATCACCAGTGCCCGTTrTT GCATCAACCCTGAGAAGACGATTTAAGCTTCACTCCCATAATTCTGGACGCAACCGCTTTTGCTGC

[0070] TrCAAGCTTTACTTTCCCTGATTCAGACTGTGTACCAGACTCAAATGCTTTGAAGTTGAGTTCGATG TTCTCAATAGCTTTATCTTTGAAAAATTGAGGCAGCTCAGTGAACAGAGACTGAAAAATTATrTCA TACGCTAAAGCACGACACTGCAGATCATTAATCACGTCTTCTACTGGCTTCATAACTTCTCCTCAAA AAACACGA

[0071] 3 xerC2 DNA ATGACTGTCCGTAAAAATCCTGCTGGCGGCTGGATTTGCGAACTCTATCCAAACGGGGCAAAAGG sequence CAAACGCA7TAGAAAGAAA7TCGCCACCAAAGGTGAGGCA7TGGCCITtGAGCAATACACAGTAC AAAACCCGTGGCAGGAAGAAAAAGAAGACAGGCGTACGTTAAAGGATCTGATTGACGCGTGGTT TAGCGCTCACGGCATAACTCTGAAAGACGGTCTCAAACGCCAGCTCGCGATGCATCATGCTITCGA GTGTATGGGGGAACCGCTCGCGCGCGATTTTGATGCGCAAATATTTTCCCGCTACCGAGAAAAAC GGTTAAAAGGTGAGTATGCCCGTtCAAACAGGGTGAAAGAGGTATCGCCTCGCACGCn’AATCTT GAACTGGCCTACTTCCGGGCGGTATTCAATGAGCTAAACCGCCTCGGAGAATGGAAGGGTGAGA ACCCGCTGAAAAATATGCGCCCATTCCGCACAGAAGAAATGGAAATGGCCTGGCTAACTCACGAC CAAATTTCGCAACTGCTCGGAGAGTGCAAAAGGCATGACCACCCTGATTTAGAAACAGTGGTAAG AATCTGTCTCGCCACTGGCGCACGCTGGTCTGAGGCCGAGAGTCTGAGAAAAAGCCAGCTCGCGA AATACAAAATCACATACACCAACACGAAAGGCAGAAAAAACCGCTCTGTCCCAATCAGCAAAGAG CTCTATGAGTCTCTGCCTGATGATAAAAAAGGCCGGTTATTTAGTGATTGTTATGGGGCGTTCCGG TCAGCTCTGGAAAGAACAGGCATCGAACTACCGGCAGGGCAACTTACCCATGTT7TACGCCACAC

[0072]

[0073] CTTCGCCAGTCACTTTATGATGAATGGTGGTAATATTCTGGTCTTGCAGCGAGTGCTCGGCCACAC Attorney Docket No. PIV-00053 WO

[0074] CGACATAAAAATGACAATGCGATATGCGCATTTCGCGCCAGATCATTTAGAAGATGCAGTCAAAT TAAATCCTTTAAGCTTTAAAACTAACAATTAA

[0075] 4 XerC2 MTVRKNPAGGWICELYPNGAKGKRIRKKFATKGEALAFEQYTVQNPWQEEKEDRRTLKDLIDAWFSA polypeptide HGITLKDGLKRQLAMHHAFECMGEPLARDFDAQIFSRYREKRLKGEYARSNRVKEVSPRTLNLELAYFR AVFNELNRLGEWKGENPLKNMRPFRTEEMEMAWLTHDQISQLLGECKRHDHPDLETWRICLATGA RWSEAESl. RKSQI. AKYKITYTNTKGRKNRSVPISKEl. YESl. PDDKKGRLFSDCYGAFRSAI. ERTGIEI. PAG QLTHVLRHTFASHFMMNGGNILVLQRVLGHTDIKMTMRYAHFAPDHLEDAVKLNPLSFKFNN

[0076] 5 xerC3 DNA ATGACGGTCAGCAAACAGAAAAATGGGAAGTGGCTGTGCGAGCTCTATCCGCAGGGACGGGAG sequence GGGCGACGTATTCGTCGGCAGTTTCCTACCAAAGGTGAAGCCGAGGCATTTGAGCGCTGGACGA AGCAAGAGGCGCAGGAGAAGCCCTGGCTTGGCGAAAAAGAGGACCGCCGTCGGTTGAGCGATC TGATTGCGCTGTGGTTTAAGTTGCATGGGCAGTCGCTGGCGGCCGGGAAATCGCGGATGGCTAA GCTGGAGATTGTCTGCCGGGGGCTCGGCGACCCACTGGCGACGGAGCTGACCGCGAAAGTGTGG GCGCACTATCGCGATCGGCGTTTGAGTGGCGAAATCGATAATGGTTACACGCGCGATCCGGCGAA CTGGAAGGTGAAGCCGGTGACGGTGAACCGTGAGCAGCAGTATCTCAGTGCGGTGTTTAACGAG CTGAGTCGGTFGGGGGAGTGGCAGTTACCGAACCCGCTCGAGGGGGTACGGACGT1TCGCGAGA AAGAGCGCGAGATGACGTGGCTGACACCGCCGCAAATCGCCACACTGCTGGCGGCCTGTGACGC GTTCGGGAATCGCGAT1TAACGCGGGTGGTGAAGGTGTGTCTTGCCACCGGCGCGCGCTGGCGC G AAG CG G AAAACCTTCAG CG CACG CAGCTTTCG G CCAATAAAATC ACGTTTGTG AAAACCAAG GG CGGCAAGAACCGTACCGTGCCCATTCCGCAGTGGTrGTATGACGAGCTTGCGCCATTGCAGGGGC AGATGTTCCAGCCGTGCTATCCGGCGTTCAGCAAAATGCTGGCGTCCACCGATATCGCACTGGCC GAGGGGCAGAAAACCCATGTGTTGCGCCATACCTTTGCCGCGCAT7TCATGACCAATGGCGGCAA TATCCTGGTACTGCAGCGTATTCTGGGTCACGCCAATATCCGTGAAACCCTGCGTTACGCCCACTT CGCGCCCGATCATCTGGAGGAAGCGGTAACCCTGAACCCGCTGGCGCATTTTAATGGCGGCAAAA TGGCGGCAGAAGTTGCATAA

[0077] 6 XerC3 MTVSKQKNGKWI. CELYPQGREGRRIRRQFPTKGEAEAFERWTKQEAQEKPWI. GEKEDRRRLSDl. IAL polypeptide WFKLHGQSLAAGKSRMAKLEIVCRGLGDPLATELTAKVWAHYRDRRLSGEIDNGYTRDPANWKVKP VTVNREQQYLSAVFNEI5RLGEWQLPNPLEGVRTFREKEREMTWl. TPPQIATLLAACDAFGNRDl. TR WKVCLATGARWREAENLQRTQLSANKITFVKTKGGKNRTVPIPQWLYDELAPLQGQMFQPCYPAFS KMLASTDIALAEGQKTHVLRHTFAAHFMTNGGNILVLQRILGHANIRETLRYAHFAPDHLEEAVTLNPL AHFNGGKMAAEVA

[0078] 7 xerD DNA ATGTC AATC AC CCTC AG G GG CG GCGTCTG GC ACTGTCATTTCGTTACG CC GTCAG G G AAAAG AAT sequence TAGACGATCTCTTGGTACGGGGGACAAAAAGCAAGCGCAGGAATTGCACGACAAGCTGAAGGCA GAAGCGTGGCGGGTGGATCAGATAGGTGATTTGCCAACAAGGACGTTTGAAGAGTGTTGCATCC GCTGGTTGCGGGAGAAAGATGATAAGCGATCTCTGGATGATGACAGGACGAAGATAGAAT1TTT CCTGCAACATTTCTCAGGACGGGATATTTCGACCATTACAGCCGAACAGGTTCATGATTCTGTGTC GAAAATGATT. AACCGTAAGCATTTGC. AGGTCTGGGA. ATCGCGGCGAGATTCCGCATTGCGGAAG GGTCAAGAGCCACCAGCATATGTGGCAAAGCCGGTTAGCCAGGCAACTAAAAGCCAGCATCTTTC TTTCATGCGATCCCTGCTTCGAGCGGCGGCCAATGACTGGGGGTGGATAAAGGCGGCCCCGGTA ATTAAAACAAAGAAACCGGTCAGTAAGCGCATTCGCTGGCTGACTCGCGATGAGGCCGAAAGGC TGATTGAGTGTATGCCGGAAAGCATAAAGCCGGTGGTGATTTTTGCACTGGCTACCGGCCTGCGC CGCTCCAACATCATTGATCTGGAGTGGCAGCAGGTCGATATGCAGAGAAAGGTTGCATGGGTAA ACCCGGAGAACGCAAAAGCGGGCAAGGCTATCGGCGTCGCTCTGAATGATACCGCATGCAGGGT TCTGAGGGATCAGATAGGTAAAAGCTCACGGTGGGTATTTGTGCATACAAAGGCAAAGCACCGG CCAGACGGCACGCTGACACCAGCAGTCAGGAAAATGCGTGTTGATGATAATCAGGCATGGAACA ITGGCCTGAAGAAAGCAGGCATCGAAGACTTCCGTmCATGACCFCCGGCACACCTGGGCGAGT TGGCTGATTCAGTCCGGCGTTCCGCTGTCCGTATTGCAGGAAATGGGAGGCTGGGAGTCAATAGA GATGGTGAGACGTIATGCGCACCTCGCGCCTAATCACCTGAGTGAGCACGCGCGGAAAATTGATG CCATTTTTGGCAACCATGACACAAATACGACACAAGGAGGAAATCAGGCTGGATTGAGACTTGCG TAA

[0079] 8 XerD MSITLRGGVWHCHFVTPSGKRIRRSLGTGDKKQAQELHDKLKAEAWRVDQIGDLPTRTFEECCIRWL polypeptide REKDDKRSLDDDRTKIEFFLQHFSGRDISTITAEQVHDSVSKMINRKHLQVWESRRDSALRKGQEPPAY VAKPVSQATKSQHLSFMRSLLRAAANDWGWIKAAPVIKTKKPVSKRIRWLTRDEAERLIECMPESIKP WIFALATGLRRSNIIDLEWQQVDMQRKVAWVNPENAKAGKAIGVALNDTACRVLRDQIGKSSRWV FVHTKAKHRPDGTLTPAVRKMRVDDNQAWNIGLKKAGIEDFRFHDLRHTWASWLIQSGVPLSVLQE

[0080]

[0081] MGGWESIEMVRRYAHLAPNHLSEHARKIDAIFGNHDTNTTQGGNQAGLRLA

[0082] A modified prophage can have a DNA sequence that has at ieast 80% sequence identity (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, Attorney Docket No. PIV-00053 WO

[0083] 99.95%, or 99.99% sequence identity) to SEQ ID NOs 9-11. For exampie, a modified

[0084] prophage can have a DNA sequence that has at least 80% sequence identity (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.95%, or 99.99% sequence identity) to any of the exemplary modified prophage genome sequences put forth in Table 2 or any of the modified prophage genome sequences disclosed herein.

[0085] Table 2. Exemplary Modified Prophage Genome Sequences.

[0086] SEQ. ID Included Prophage Modified prophage sequence

[0087] NO: in Modification

[0088] Bacterial

[0089] Strain

[0090] o 5812- xerD

[0091] 7767 (recombinase)

[0092] deletion

[0093] CTTCAGCAATTTATGATCAATAAAATACGCGTGCGCGGTGCGCGATTTAATCTGTGTGCGCTCTGCTTCCCGCGTCAGGCC

[0094]

[0095] Attorney Docket No. PIV-00053 WO

[0096]

[0097] Attorney Docket No. PIV-00053 WO

[0098]

[0099] Attorney Docket No. PIV-00053 WO

[0100]

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[0102]

[0103] Attorney Docket No. PIV-00053 WO

[0104] AATCACGTAAGGAAATCGCCAGAGAAGGCTACGAAATCGACGGACAGCGCTACAGCATCAAATTCAACGTTGTTCCGAA

[0105] TCAACTTGCCGATATCAAAATGACGTTAAACGCCTGCGACATGAAGTACAGCAAGCCAAATGTCTACGGCAAGTCAACGT

[0106]

[0107] Attorney Docket No. PIV-00053 WO

[0108]

[0109] Attorney Docket No. PIV-00053 WO

[0110]

[0111] Attorney Docket No. PIV-00053 WO

[0112]

[0113] Attorney Docket No. PIV-00053 WO

[0114]

[0115] Attorney Docket No. PIV-00053 WO

[0116]

[0117] Attorney Docket No. PIV-00053 WO

[0118]

[0119] Attorney Docket No. PIV-00053 WO

[0120] CCATGGATATGGCAGAGGATGTTATTCAGGTAACTGATCCGGTATCAGGCATCGTGTTTGATATTGCCGTTTACCGCCAG

[0121] GCATTAACAGACGCTCAAATGACTGATGTTCGCCGGTACATGGGGTATCAGCTTAGTGGTACTACTATGCCAATAACTGC

[0122]

[0123] Attorney Docket No. PIV-00053 WO

[0124]

[0125] Attorney Docket No. PIV-00053 WO

[0126] A

[0127]

[0128] Attorney Docket No. PIV-00053 WO

[0129]

[0130] In some cases, the modified prophage described herein can contain fewer nucleotides (i.e., less DNA) than the corresponding unmodified prophage DNA sequence. For example, a modified prophage as described herein can include from about 51% to about 99.99% of the nucleotide sequence of the prophage (e.g., from about 55% to about 99.99%, from about 60% to about to about 99.99%, from about 65% to about 99.99%, from about 70% to about 99.99%, from about 75% to about 99.99%, from about 80% to about 99.99%, from about 85% to about 99.99%, from about 90% to about 99.99%, from about 95% to about 99.99%, from about 96% to about 99.99%, from about 97% to about 99.99%, from about 98% to about 99.99%, from about 99% to about 99.99%, from about 51% to about 99%, from about 51% to about 98%, from about 51% to about 97%, from about 51% to about 96%, from about 51% to about 95%, from about 51% to about 90%, from about 51% Attorney Docket No. PIV-00053 WO

[0131] to about 85%, from about 51% to about 80%, from about 51% to about 75%, from about 51% to about 70%, from about 51% to about 65%, from about 51% to about 60%, from about 51% to about 55%, from about 60% to about 85%, from about 65% to about 90%, or from about 70% to about 85%). In some cases, a modified prophage can include from about 51% to about 99.99% of the nucleotide sequence of the prophage.

[0132] Genetically Modified Bacteria Comprising Modified Prophage

[0133] Many different microbial species can be modified to produce a bacterium that includes a modified prophage as described herein. For example, a bacterium used in the methods and / or compositions described herein can comprise one bacterial species or a plurality of different bacterial taxa in combination. For example, a bacterium or bacterial host cell genome containing a modified prophage as described herein and to be used in any of the methods or compositions described herein can be selected from a variety of bacterial species that contain prophage. In some cases, host bacterial cells genomes that contain a modified prophage as described herein are more stable and produce more product when exposed to an environmental stress, such as nutrient stress, salt stress, osmotic stress, cold stress, heat stress, excess UV radiation, the presence of antibiotics, and DNA damage. In some cases, host bacterial cell genomes that contain a modified prophage as described herein have a decreased rate or likelihood of dying ("crashing") when fermenting over a 1 hour, 8 hour, 12 hour, 18 hour, 24 hour, 48 hour, 1 week, 2 week, or 1 month period when grown in bacterial culture medium.

[0134] A modified bacterium can have certain traits, such as desirable traits. In some cases, a modified bacterium has an increased cell growth rate in comparison to a corresponding bacterium lacking the prophage. Growth rate can be measured in any appropriate manner. For example, growth rate can be measured in a liquid culture medium, in a soil sample (e.g., a soil sample in which a modified bacterium has been deposited), or in a field (e.g., a field in which a modified bacterium has been deposited). In some cases, modified bacterial cells are more likely to contain an intact gene of interest over an extended period of time as compared to a corresponding bacterial cell lacking the genetic modification of the modified bacterial cell. In some cases, products made with any of the modified bacterial cells described remain shelf-stable for a longer period of time as compared to a correspond bacterial cell lacking the genetic modification of the modified bacterial cell. In another embodiment, products made with any of the modified bacterial cells described herein Attorney Docket No. PIV-00053 WO

[0135] remain more stable and are able to produce product longer when the modified bacterial ceils are applied to crops, furrows, seeds, or soil, in comparison to a product made with a correspond bacterial cell lacking the genetic modification of the modified bacterial cell.

[0136] In some cases, the methods and / or compositions disclosed herein may utilize a bacterial species that is a Gram-positive bacterium or a Gram-negative bacterium.

[0137] Examples of Gram-positive microbes include Paenibacillus polymixa, Paenibacillus riograndensis, Paenibacillus sp., Frankia sp., Heliobacterium sp., Heliobacterium chlorum, Heliobacillus sp., Heliophilum sp., Heliorestis sp., Clostridium acetobutylicum, Clostridium sp., Mycobacterium jlaum, Mycobacterium sp., Arthrobacter sp., Agromyces sp., Corynebacterium autitrophicum, Corynebacterium sp., Micromonspora sp., Propionibacteria sp., Streptomyces sp., and Microbacterium sp.

[0138] By way of example, the bacterium may include Proteobacteria (such as Pseudomonas, Enterobacter, Stenotrophomonas, Burkholderia, Paraburkholderia, Rhizobium, Herbaspirillum, Pantoea, Serratia, Rahnella, Azospirillum, Azorhizobium, Azotobacter, Duganella, Delftia, Bradyrhizobiun, Sinorhizobium and Halomonas), Firmicutes (such as Bacillus, Paenibacillus, Lactobacillus, Mycoplasma, and Acetabacterium), and Actinobacteria (such as Streptomyces, Rhodacoccus, Microbacterium, and Curtobacterium).

[0139] Bacteria that can be produced by the methods disclosed herein include In some examples, bacteria used according to the methods of the disclosure can be a member of one or more of the following genera: Achromobacter, Acidithiobacillus, Acidovorax, Acidovoraz, Acinetobacter, Actinoplanes, Adlercreutzia, Aerococcus, Aeromonas, Afipia, Agromyces, Ancylobacter, Arthrobacter, Atopostipes, Azospirillum, Bacillus, Bdellovibrio, Beijerinckia, Bosea, Bradyrhizobium, Brevibacillus, Brevundimonas, Burkholderia, Candidatus, Caulobacter, Cellulomonas, Cellvibrio, Chryseobacterium, Citrobacter, Clostridium, Coraliomargarita, Corynebacterium, Cupriavidus, Curtobacterium, Curvibacter, Deinococcus, Delftia, Desemzia, Devosia, Dokdonella, Dyella, Enhydrobacter, Enterobacter, Enterococcus, Erwinia, Escherichia, Escherichia, Shigella, Exiguobacterium, Ferroglobus, Filimonas, Finegoldia, Flavisolibacter, Flavobacterium, Frigoribacterium, Gluconacetobacter, Hafnia, Halobaculum, Halomonas, Haloredivivus, Halosimplex, Herbaspirillum, Hymenobacter, Klebsiella, Kocuria, Kosakonia, Lactobacillus, Leclercia, Lentzea, Luteibacter, Luteimonas, Massilia, Mesorhizobium, Methylobacterium, Microbacterium, Micrococcus, Microvirga, Mycobacterium, Neisseria, Nocardia, Oceanibaculum, Ochrobactrum, Okibacterium, Oligotropha, Oryzihumus, Oxalophagus, Paenibacillus, Panteoa, Pelomonas, Perlucidibaca, Attorney Docket No. PIV-00053 WO

[0140] Plantibacter, Poiynucleobacter, Propionibacterium, Propioniciclava, Pseudoclavibacter, Pseudomonas, Pseudonocardia, Pseudoxanthomonas, Psychrobacter, Ralstonia, Rahnella, Rheinheimera, Rhizobium, Rhodococcus, Rhodopseudomonas, Roseateles, Ruminococcus, Sebaldella, Sediminibacillus, Sediminibacterium, Serratia, Shigella, Shinella, Sinorhizobium, Sinosporangium, Sphingobacterium, Sphingomonas, Sphingopyxis, Sphingosinicella, Staphylococcus, Stenotrophomonas, Strenotrophomonas, Streptococcus, Streptomyces, Stygiolobus, Sulfurisphaera, Tatumella, Tepidimonas, Thermomonas, Thiobacillus, Variovorax, WPS-2 genera incertae sedis, Xanthomonas, and Zimmermannella.

[0141] In some cases, the bacteria can be selected from the group consisting of:

[0142] Azotobacter sp., Bradyrhizobium sp., Klebsiella sp., Kosakonia sp., and Sinorhizobium sp. In some cases, the bacteria can be selected from the group consisting of: Azotobacter sp., Bradyrhizobium sp., Klebsiella sp., and Sinorhizobium sp. In some cases, the bacteria may be selected from the group consisting of: Azotobacter vinelandii, Bradyrhizobium japonicum, Klebsiella pneumoniae, and Sinorhizobium meliloti. In some cases, the bacteria can be of the genus Azospirillum, for example, Azospirillum palustre. In some cases, the bacteria may be of the genus Enterobacter or Rahnella. In some cases, the bacteria may be of the genus Frankia, or Clostridium. Examples of bacteria of the genus Clostridium include, but are not limited to, Clostridium acetobutilicum, Clostridium pasteurianum, Clostridium beijerinckii, Clostridium perfringens, and Clostridium tetani. In some cases, the bacteria can be of the genus Kosakonia, for example, Kosakonia arachidis and Kosakonia sacchari. In some cases, the bacteria may be of the genus Paenibacillus, for example, Paenibacillus azotofixans, Paenibacillus borealis, Paenibacillus durus, Paenibacillus macerans, Paenibacillus polymyxa, Paenibacillus alvei, Paenibacillus amylolyticus, Paenibacillus campinasensis, Paenibacillus chibensis, Paenibacillus glucanolyticus, Paenibacillus illinoisensis, Paenibacillus larvae subsp. Larvae, Paenibacillus larvae subsp. Pulvifaciens, Paenibacillus lautus, Paenibacillus macerans, Paenibacillus macquariensis, Paenibacillus macquariensis, Paenibacillus pabuli, Paenibacillus peoriae, or Paenibacillus polymyxa.

[0143] In some embodiments, the methods of the disclosure may utilize a bacterial species selected from at least one of the following genera: Enterobacter, Klebsiella, Kosakonia, and Rahnella. In some cases, one or more bacterial species selected from the following genera are utilized: Enterobacter, Klebsiella, Kosakonia, and Rahnella. In some cases, one or more bacterial species individually selected from the following genera are utilized: Enterobacter, Klebsiella, Kosakonia, and Rahnella. In some cases, the bacterial species utilized can be one Attorney Docket No. PIV-00053 WO

[0144] or more of: Enterobacter sacchari, Klebsiella variicola, Kosakonia sacchari, Kosakonia arachidis and Rahnella aquatilis.

[0145] In some cases, the bacterium used in any of the methods or compositions described herein is a nitrogen-fixing bacterium. The bacterium used in methods and compositions of this disclosure may include nitrogen-fixing bacterial consortia of two or more species. In some cases, one or more bacterial species of the bacterial consortia may be capable of fixing nitrogen. In some cases, one or more species of the bacterial consortia may facilitate or enhance the ability of other bacteria to fix nitrogen. The bacteria which fix nitrogen and the bacteria which enhance the ability of other bacteria to fix nitrogen may be the same or different. In some cases, a bacterial strain may be able to fix nitrogen when in combination with a different bacterial strain, or in a bacterial consortium, but may be unable to fix nitrogen in a monoculture. Examples of bacterial genera that can be found in a nitrogenfixing bacterial consortium include, but are not limited to, Herbaspirillum, Azospirillum, Enterobacter, and Bacillus.

[0146] Any of the modified bacteria described herein can also include additional genetic modifications. For example, the additional genetic modifications can increase the nitrogenfixing capacity of the modified bacteria. In some cases, the additional genetic modifications can include modifications in genes involved in nitrogen fixation or nitrogen assimilation (e.g., n / / A, nifL, ntrB, ntrC, polynucleotide encoding glutamine synthetase, glnA, glnB, glnK, draT, amtB, polynucleotide encoding glutaminase, glnD, glnE, nifJ, nifH, nifD, nifK, nifY, nifE, nifU, nifS, nifV, nifW, nifZ, nifM, nifF, nifB, and nifQ). Exemplary modifications are disclosed in WO / 2017 / 062412, WO / 2017 / 011602, WO / 2018 / 132774, WO / 2019 / 084059, WO / 2019 / 084342, WO 2020 / 219932, W02020 / 191201, WO 2021 / 221690, WO 2021 / 231449, WO 2023 / 278804, and WO 2021 / 222567, each of which is incorporated in its entirety herein.

[0147] Any of the bacterial strains in Table 3 can be modified to contain any of the modified prophages described herein. For example, Table 3 lists the deposit information for exemplary bacterial strains deposited with National Center for Marine Algae and Microbiota (NCMA) or ATCC. Each of the deposits was made under the provisions of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purpose of Patent Procedure and the Regulations (Budapest Treaty).

[0148] Table 3: Microorganisms Deposited under the Budapest Treaty Attorney Docket No. PIV-00053 WO

[0149] Strain

[0150] Designation

[0151] Accession

[0152] Depository (multiple Taxonomy Date of Deposit Number

[0153] designations

[0154] indicated)

[0155] CI006,

[0156] NCMA Kosakonia sacchari (WT) 201701001 January 06, 2017

[0157] PBC6.1,6

[0158] NCMA Ci019,19 Rahnella aquatilis (WT) 201701003 January 06, 2017 NCMA CM029, 6-412 Kosakonia sacchari 201701002 January 06, 2017

[0159] 6-403

[0160] NCMA Kosakonia sacchari 201708004 August 11, 2017

[0161] CM037

[0162] 6-404,

[0163] NCMA CM38, Kosakonia sacchari 201708003 August 11, 2017

[0164] PBC6.38

[0165] CM094,

[0166] NCMA 6-881, Kosakonia sacchari 201708002 August 11, 2017

[0167] PBC6.94

[0168] CI137, 137,

[0169] NCMA Klebsiella variicola (WT) 201708001 August 11, 2017

[0170] PB137

[0171] NCMA 137-1034 Klebsiella variicola 201712001 December 20, 2017 NCMA 137-1036 Klebsiella variicola 201712002 December 20, 2017 ATCC 6-2425 Kosakonia sacchari PTA-126575 December 23, 2019 ATCC 6-2634 Kosakonia sacchari PTA-126576 December 23, 2019 ATCC 137-1968 Klebsiella variicola PTA-126577 December 23, 2019 ATCC 137-2219 Klebsiella variicola PTA-126578 December 23, 2019 ATCC 137-2237 Klebsiella variicola PTA-126579 December 23, 2019 ATCC 137-2285 Klebsiella variicola PTA-126580 December 23, 2019 ATCC 41 Paenibacillus polymyxa (WT) PTA-126581 December 23, 2019 ATCC Paraburkholderia tropica PTA-126582 December 23, 2019

[0172] 8

[0173] (WT)

[0174] ATCC Herbaspirillum aquaticum PTA-126583 December 23, 2019

[0175] 3069

[0176] (WT)

[0177] ATCC 137-2253 Klebsiella variicola PTA- 126740 March 25, 2020 ATCC 137-3896 Klebsiella variicola PTA- 126741 March 25, 2020 ATCC 137-3890 Klebsiella variicola PTA- 126749 March 25, 2020 ATCC 6-5687 Kosakonia sacchari PTA- 126743 March 25, 2020 ATCC 5812 Kosakonia oryzendophytica PTA- 127355 August 8, 2022

[0178]

[0179] ATCC 1661-5402 Kosakonia arachidis PTA- 127097 July 20, 2021

[0180] Methods of Grounding a Prophage

[0181] As described herein, prophages can be grounded by modifying the prophage. In

[0182] some cases, methods of grounding a prophage include genetically modifying a prophage in a bacterium to completely delete, partially delete, or inactivate a nucleic acid encoding a recombinase, an attL DNA sequence, an attR DNA sequence, or any combination thereof

[0183] from the prophage sequence of interest. In some cases, methods of grounding a prophage Attorney Docket No. PIV-00053 WO

[0184] include genetically modifying a prophage in a host bacterial cell genome to obtain any of the modified prophages described herein.

[0185] In some cases, prophages can be grounded using the following general method. A bacterium can be previously identified (e.g., in the scientific literature ora previously performed analysis) or identified as comprising a genome having a prophage. For example, a bacterium can be identified as comprising a genome having a prophage using any appropriate bioinformatic analysis as described herein. For example, a bacterium can be identified as comprising a genome having a prophage using whole genome sequencing followed by read coverage and bioinformatic analyses, or any sequencing method that generates genomic data for the location in the genome suspected of having a prophage. Whole genome sequencing data can be used to identify the 5' and 3' edges of the prophage as well as putative recombinases. Prophage edges and nucleic acids encoding putative recombinases can be genetically modified (e.g., deleted) yielding a prophage that is grounded. Attachment sites are at the edges of the prophage. Thus, deleting the areas at the 5' or 3' edges of the prophage will also delete the attL and attR sites. In some cases, the deletion bridges the host genome and the prophage genome. In some cases, multiple prophages are identifed in the bacterial genome and each prophage is grounded.

[0186] Any appropriate whole genome sequencing method can be used. For example, whole genome sequencing can use the method described in Example 3.

[0187] Any appropriate read coverage analysis tool can be used to determine read coverage, such as using a SAMtool mpileup function. For example, read coverage can be determined as described in Example 4.

[0188] Any appropriate bioinformatic analysis can be used to identify the 5' and 3' edges of the prophage, such as PhiSPy or VirSorter2. For example, the 5' and 3' edges of a prophage can be identified as described in Example 4.

[0189] Any appropriate bioinformatic analysis can be used to identify putative phage recombinases. For example, gene function can be predicted using any appropriate annotation program, such as DRAMM. For example, putative phage recombinases can be identified as described in Example 4. In some cases, one recombinase in a target prophage can be identified. In some cases, multiple recombinases in a target prophage can be identified. In some cases, multiple recombinases in multiple prophages within a host bacterial cell genome can be identified. Attorney Docket No. PIV-00053 WO

[0190] The process of generalized grounding via edge trimming (via removal of the 5'or 3' edges of the prophage genome overlapping with the host genome resulting in the deletion of attL and attR) and phage recombinase deletion is showcased here in a strain of Kosakonia oryzendophytica strain 5812 (FIG, 3). This process involves minimal deletion of DNA sequences in comparison to a complete prophage deletion (i.e., a deletion of the entire prophage genome from the host bacterial genome).

[0191] In some cases, modifying the prophage includes modifying an attL DNA sequence of the prophage. For example, an attL DNA sequence of the prophage can be completely deleted, partially deleted, or inactivated such that the prophage is inhibited from excising from the host bacterial cell genome. Any suitable genetic modification that results in the inactivation of an attL DNA sequence of the prophage can be used. For example, an attL DNA sequence of the prophage can be completely deleted, partially deleted, or inactivated such that the prophage is inhibited from excising from the host bacterial cell genome. For example, the complete attL DNA sequence of the prophage can be deleted from the host bacterial cell genome. In some cases, a part of an attL DNA sequence of the prophage can be deleted from the host bacterial cell genome to generate a partial deletion. For example, a partial deletion of an attL DNA sequence can remove the active portion of the attL DNA sequence (e.g., the DNA sequence that recognized recombinase to excise the prophage). In some cases, modifying the prophage includes modifying a single nucleotide (e.g., a point mutation) in an attL DNA sequence of the prophage causing the attL DNA sequence of the prophage to be inactivated.

[0192] In some cases, modifying the prophage includes modifying an attR DNA sequence of the prophage. For example, an attR DNA sequence of the prophage can be completely deleted, partially deleted, or inactivated such that the prophage is inhibited from excising from the host bacterial cell genome. Any suitable genetic modification that results in the inactivation of an attR DNA sequence of the prophage can be used. For example, an attR DNA sequence of the prophage can be completely deleted, partially deleted, or inactivated such that the prophage is inhibited from excising from the host bacterial cell genome. For example, the complete attR DNA sequence of the prophage can be deleted from the host bacterial cell genome. In some cases, a part of an attR DNA sequence of the prophage can be deleted from the host bacterial cell genome to generate a partial deletion. For example, a partial deletion of an attR DNA sequence can remove the active portion of the attR DNA sequence (e.g., the DNA sequence that recognized recombinase to excise the prophage). In Attorney Docket No. PIV-00053 WO

[0193] some cases, modifying the prophage includes modifying a single nucleotide (e.g., a point mutation) in an attR DNA sequence of the prophage causing the attR DNA sequence of the prophage to be inactivated.

[0194] In some cases, modifying the prophage includes modifying a nucleic acid sequence encoding a recombinase polypeptide of the prophage. Any suitable genetic modification that results in the inactivation of a nucleic acid sequence encoding a phage recombinase polypeptide can be used. For example, a nucleic acid sequence encoding a recombinase polypeptide of the prophage can be completely deleted, partially deleted, or inactivated such that the prophage is inhibited from excising from the host bacterial cell genome. For example, the complete nucleic acid sequence encoding a phage recombinase polypeptide can be deleted from the host bacterial cell genome. In some cases, a part of a nucleic acid sequence encoding a phage recombinase polypeptide can be deleted form the host bacterial cell genome to generate a partial deletion. For example, a partial deletion of a nucleic acid sequence encoding a phage recombinase polypeptide can remove the active portion of the recombinase sequence (e.g., the DNA sequence that recognized the attL and / or attR DNA sequences to excise the prophage). In some cases, modifying the prophage includes modifying a single nucleotide (e.g., a point mutation) in the nucleic acid sequence encoding a phage recombinase polypeptide causing the recombinase polypeptide to be inactivated.

[0195] In some cases, modifying the prophage includes modifying one or more nucleic acid sequences encoding one or more phage recombinase polypeptides. The one or more nucleic acid sequences (e.g., two, three, four, five, six or more nucleic acid sequences) encoding one or more phage recombinase polypeptides (e.g., two, three, four, five, six or more phage recombinase polypeptides) can be from a single prophage or can be from multiple prophages identified or previously identified as being in the host bacterial cell genome. In some cases, modifying the prophage includes modifying one or more nucleic acid sequences encoding one or more recombinase polypeptides within a target prophage. In some cases, modifying the prophage includes modifying one or more nucleic acid sequences encoding one or more recombinase polypeptides within multiple prophages in the host bacterial cell genome. In some cases, modifying the prophage includes modifying all of the nucleic acid sequences encoding phage recombinase polypeptides. In some cases, modifying the prophage includes deleting (e.g., completely deleting or partially deleting) one or more nucleic acid sequences encoding one or more phage recombinase polypeptides. In some cases, modifying the prophage includes deleting (e.g., completely deleting or partially Attorney Docket No. PIV-00053 WO

[0196] deleting) one or more nucleic acid sequences encoding one or more recombinase polypeptides within a target prophage. In some cases, modifying the prophage includes deleting (e.g., completely deleting or partially deleting) one or more nucleic acid sequences encoding one or more recombinase polypeptides within multiple prophage in the host bacterial cell genome. In some cases, modifying the prophage includes deleting (e.g., completely deleting or partially deleting) all of the nucleic acid sequences encoding phage recombinase polypeptides. In some cases, modifying the prophage includes inactivating one or more nucleic acid sequences encoding one or more phage recombinase polypeptides. In some cases, modifying the prophage includes inactivating one or more nucleic acid sequences encoding one or more recombinase polypeptides within a target prophage. In some cases, modifying the prophage includes inactivating one or more nucleic acid sequences encoding one or more recombinase polypeptides within multiple prophages in the host bacterial cell genome. In some cases, modifying the prophage includes inactivating all of the nucleic acid sequences encoding recombinase polypeptides. In some cases, identifying and modifying multiple nucleic acids encoding a recombinase polypeptide can use the method set forth in Examples 1-10. As such, in some cases, the bacterial cell genome contains a a prophage genome that does not encode an active phage recombinase polypeptide. In some cases, the bacterial cell genome contains multiple prophage genomes and does not include a nucleic acid sequence encoding an active recombinase polypetide of the prophage.

[0197] In some cases, modifying the genetically modified bacteria results in a competitive fitness advantage compared to strains in which prophages were completely removed ('knocked out'). Genetically modified bacteria can be more successful (e.g., have faster growth rates, or reach higher yields) when under environmental stress as compared to corresponding genetically modified bacteria with complete prophage deletion. In addition, genetically modified bacteria as described herein can be better suited to large-scale commercial production. In addition, genetically modified bacteria can be more competitive and more successful when administered to an outdoor environment, such as administration in crops or in soil in which crops are gown, as a soil microbe to improve soil or allow better nitrogen availability to plants.

[0198] As shown herein in FIG. 4 and Example 10, genetically modified bacteria that included at least part of a prophage showed an increase in competitive fitness compared to genetically modified bacteria that lacked the entire genome of the corresponding prophage. Attorney Docket No. PIV-00053 WO

[0199] Furthermore, the genetic modifications that ground the prophage did not have a measurable impact on product performance (in this case, as measured by fixed nitrogen secretion (FIG.

[0200] 4)).

[0201] In some cases, a minimal amount of DNA modification (e.g., deletion) is performed to leave as much potential beneficial DNA in the bacterial cell as possible. For example, in some cases, less than 15% of the host bacterial cell genome is deleted (e.g., from about 0.01% to about 10%, inclusive of the prophage). In some cases, less than 15%, 20%, 25% or 30% of the prophage genome is deleted (e.g., from about 0.01% to about 30% of the prophage genome). In some cases, methods disclosed herein can include determining potential deleteriously impact of modifications on microbial performance. In some embodiments, the microbial performance is the delivery of atmospheric nitrogen-derived nitrogen to a crop. In some cases, a genetically modified bacterium did not exhibit a decrease in production of a commercially-relevant product (e.g., ammonium as assessed by a nitrogen-fixation assay). For example, a genetically modified bacterium producing a commercially-relevant product, such as fixed-nitrogen, produced the commercially-relevant product at levels substantially similar to a corresponding bacterium lacking the genetic modifications of the genetically modified bacterium. As shown in Example 11 and in FIG. 5, the impact of the prophage grounding procedure on product performance was measured, using the grounded strain of Kosakonia oryzendophytica strain 5812. The product performance for this strain, that is, the ability to fix nitrogen and secrete ammonium, was not inhibited or decreased by the genetic modifications that were performed to disable the prophage excision, when compared to corresponding strains containing the prophage sequences (FIG. 5).

[0202] Grounding prophage in a host bacterial cell genome can also increase culture stability of a bacterial culture of a plurality of bacteria having a prophage as compared to a level of culture stability in a bacterial culture comprising a plurality of the corresponding bacterium lacking the prophage modification. Methods of increasing culture stability of a bacterial culture include all of the above-described methods of grounding a prophage. For example, methods of increasing culture stability of a bacterial culture include genetically modifying a prophage in a bacterium to completely delete, partially delete, or inactivate a nucleic acid encoding a recombinase, an attL DNA sequence, an attR DNA sequence, or any combination thereof from the prophage sequence of interest. In some cases, methods of increasing culture stability of a bacterial culture include genetically modifying a prophage in Attorney Docket No. PIV-00053 WO

[0203] a host bacterial cell genome to obtain any of the modified prophages described herein. In some cases, culture stability is measured by assessing prophage activation after treatment with prophage activating conditions (e.g., mitomycin C or late stationary phase growth). In some cases, increased culture stability includes a decrease in prophage activation after treatment with prophage activating conditions (e.g., mitomycin C or late stationary phase growth). In some cases, methods of increasing culture stability of a bacterial culture include determining if a host bacterial cell genome that contains a modified prophage as described herein has a decreased rate or likelihood of dying when fermenting (called culture crashing) over a 1 hour, 8 hour, 12 hour, 18 hour, 24 hour, 48 hour, 1 week, 2 week, or 1 month period when grown in bacterial culture medium. Any appropriate method can be used to determine if a host bacterial cell genome that contains a modified prophage has a decreased rate or likelihood of culture crashing.

[0204] Grounding prophage in a host bacterial cell genome can also decrease the likelihood of replication of a prophage. Methods of decreasing the likelihood of replication of a prophage include all of the above-described methods of grounding a prophage. For example, methods of decreasing the likelihood of replication of a prophage include genetically modifying a prophage in a bacterium to completely delete, partially delete, or inactivate a nucleic acid encoding a phage recombinase, an attL DNA sequence, an attR DNA sequence, or any combination thereof from the prophage sequence of interest. In some cases, methods of decreasing the likelihood of replication of a prophage include genetically modifying a prophage in a host bacterial cell genome to obtain any of the modified prophages described herein. In some cases, the likelihood of prophage replication is measured by whole genome sequencing after treatment with prophage activating conditions. In some cases, whole genome sequencing after treatment with prophage activating conditions shows no prophage activation. In some cases, methods of decreasing the likelihood of replication of a prophage include determining if a host bacterial cell genome that contains a modified prophage as described herein have a decreased rate or likelihood of dying when fermenting (called "culture crashing") over a 1 hour, 8 hour, 12 hour, 18 hour, 24 hour, 48 hour, 1 week, 2 week, or 1 month period when grown in bacterial culture medium. Any appropriate method can be used to determine if a host bacterial cell genome that contains a modified prophage has a decreased rate or likelihood of culture crashing. Attorney Docket No. PIV-00053 WO

[0205] Grounding prophage in a host bacterial cell genome can also inhibit or prevent excision of a prophage from a host bacterial cell genome. Methods of inhibiting or preventing excision of a prophage from a host bacterial cell genome include all of the abovedescribed methods of grounding a prophage. For example, methods of inhibiting or preventing excision of a prophage from a host bacterial cell genome include genetically modifying a prophage in a bacterium to completely delete, partially delete, or inactivate a nucleic acid encoding a phage recombinase, an attL DNA sequence, an attR DNA sequence, or any combination thereof from the prophage sequence of interest. In some cases, methods of inhibiting or preventing excision of a prophage from a host bacterial cell genome include genetically modifying a prophage in a host bacterial cell genome to obtain any of the modified prophages described herein. In some cases, methods of inhibiting or preventing excision of a prophage from a host bacterial cell genome include growing a bacterial culture of any of the modified bacteria described herein in a condition that activates the prophage (e.g., mitomycin C or late stationary phase growth) and determining a lack of prophage replication. In some cases, methods of inhibiting or preventing excision of a prophage from a host bacterial cell genome include determining if a host bacterial cell genome that contains a modified prophage as described herein have a decreased rate or likelihood of dying when fermenting (called "culture crashing") over a 1 hour, 8 hour, 12 hour, 18 hour, 24 hour, 48 hour, 1 week, 2 week, or 1 month period when grown in bacterial culture medium. Any appropriate method can be used to determine if a host bacterial cell genome that contains a modified prophage has a decreased rate or likelihood of culture crashing.

[0206] Grounding prophage in a host bacterial cell genome can also result in an impaired ability to lyse a host bacterial cell. Methods of impairing the ability of a prophage to lyse a host bacterial cell include all of the above-described methods of grounding a prophage. For example, methods of impairing the ability of a prophage to lyse a host bacterial cell include genetically modifying a prophage in a bacterium to completely delete, partially delete, or inactivate a nucleic acid encoding a phage recombinase, an attL DNA sequence, an attR DNA sequence, or any combination thereof from the prophage sequence of interest. In some cases, methods of impairing the ability of a prophage to lyse a host bacterial cell include genetically modifying a prophage in a host bacterial cell genome to obtain any of the modified prophages described herein. In some cases, methods of impairing the ability of a prophage to lyse a host bacterial cell include determining if a host bacterial cell genome that contains a modified prophage as described herein have a decreased rate or likelihood of Attorney Docket No. PIV-00053 WO

[0207] dying when fermenting (caiied "culture crashing") over a 1 hour, 8 hour, 12 hour, 18 hour, 24 hour, 48 hour, 1 week, 2 week, or 1 month period when grown in bacterial culture medium. Any appropriate method can be used to determine if a host bacterial cell genome that contains a modified prophage has a decreased rate or likelihood of culture crashing.

[0208] Methods of Increasing an Amount of Atmospheric Derived Nitrogen in a Plant in a Field Also provided herein are methods of increasing an amount of atmospheric derived nitrogen in a plant in a field, the method comprising contacting a soil, a plant, or a plant seed with a plurality of the modified bacteria containing modified prophages as described herein. In some cases, the plurality of modified bacteria are coated onto the plant seed. In some cases, the plurality of modified bacteria are applied into furrows in which seeds of the plant are planted. Examples of such methods are disclosed in WO / 2017 / 062412, WO / 2017 / 011602, WO / 2018 / 132774, WO / 2019 / 084059, WO / 2019 / 084342, WO 2020 / 219932, W02020 / 191201, WO 2021 / 221690, WO 2021 / 231449, WO 2023 / 278804, and WO 2021 / 222567, each of which is incorporated in its entirety herein.

[0209] The compositions comprising the modified bacteria described herein may be coated onto the surface of a seed. Examples of compositions may include seed coatings for commercially important agricultural crops, for example, sorghum, canola, tomato, strawberry, barley, rice, maize, and wheat. Examples of compositions can also include seed coatings for corn, soybean, canola, sorghum, potato, rice, vegetables, cereals, and oilseeds. Seeds as provided herein can be genetically modified organisms (GMO), non-GMO, organic, or conventional. In some examples, compositions may be sprayed on the plant aerial parts, or applied to the roots by inserting into furrows in which the plant seeds are planted, watering to the soil, or dipping the roots in a suspension of the composition. In some examples, compositions may be dehydrated in a suitable manner that maintains cell viability and the ability to artificially inoculate and colonize host plants. The modified bacteria may be present in compositions at a concentration of between about 105, 106, 107, 108, to 1010, 1011, or 1012CFU / ml. In some examples, compositions may be supplemented with trace metal ions, such as molybdenum ions, iron ions, manganese ions, or combinations of these ions. Some examples of compositions may also be formulated with a carrier, such as beta-glucan, carboxymethyl cellulose (CMC), bacterial extracellular polymeric substance (EPS), sugar, animal milk, or other suitable carriers. In some examples, peat or planting materials can be used as a carrier, or biopolymers in which a composition is entrapped in the biopolymer can Attorney Docket No. PIV-00053 WO

[0210] be used as a carrier. The compositions comprising the modified bacteria described herein can improve plant traits, such as promoting plant growth, maintaining high chlorophyll content in leaves, increasing fruit or seed numbers, and increasing fruit or seed unit weight.

[0211] Compositions comprising a seed coated with one or more modified bacteria described herein are also contemplated. The seed coating can be formed by mixing the bacterial population with a porous, chemically inert granular carrier. Alternatively, the compositions may be inserted directly into the furrows into which the seed is planted or sprayed onto the plant leaves or applied by dipping the roots into a suspension of the composition. An effective amount of the composition can be used to populate the sub-soil region adjacent to the roots of the plant with viable bacterial growth or populate the leaves of the plant with viable bacterial growth. In general, an effective amount is an amount sufficient to result in plants with improved traits (e.g., a desired level of nitrogen fixation). Plant Species

[0212] Any of the methods described herein can be used to increase an amount of atmospheric derived nitrogen in any plants that have economic, social and / or environmental value, such as food crops, fiber crops, oil crops, plants in the forestry or pulp and paper industries, feedstock for biofuel production and / or ornamental plants. Non-limiting examples of crop plants include maize, rice, wheat, barley, sorghum, millet, oats, rye triticale, buckwheat, sweet corn, sugar cane, onions, tomatoes, strawberries, and asparagus. For example, plants can be in the genus Hordeum, Oryza, Zea, and Triticeae. In some cases, any of the methods described herein can be used to increase an amount of atmospheric derived nitrogen in cereal plants. Non-limiting examples of cereal plants include corn plants, canola plants, sorghum plants, wheat plants, and sunflower plants.

[0213] In some cases, any of the methods described herein can be used to increase an amount of atmospheric derived nitrogen in plants that may be useful or interesting for agriculture, horticulture, biomass for the production of biofuel molecules and other chemicals, and / or forestry. Some non-limiting examples of these plants include pineapple, bamboo, banana, coconut, lily, grass peas and grass; and dicotyledonous plants, such as, for example, peas, alfalfa, tomatillo, melon, chickpea, chicory, clover, kale, lentil, soybean, tobacco, potato, sweet potato, radish, cabbage, rape, apple trees, grape, cotton, sunflower, thale cress, canola, citrus (including orange, mandarin, kumquat, lemon, lime, grapefruit, tangerine, tangelo, citron, and pomelo), pepper, bean, lettuce, Panicum virgatum (switch), Sorghum bicolor (sorghum, sudan), Miscanthus giganteus (miscanthus), Saccharum sp. Attorney Docket No. PIV-00053 WO

[0214] (energycane), Populus balsamifera (poplar), Zea mays (corn), Glycine max (soybean), Brassica napus (canola), Triticum aestivum (wheat), Gossypium hirsutum (cotton), Oryza sativa (rice), Helianthus annuus (sunflower), Medicago sativa (alfalfa), Beta vulgaris (sugar beet), Pennisetum glaucum (pearl millet), Panicum spp. Sorghum spp., Miscanthus spp., Saccharum spp., Erianthus spp., Populus spp., Secale cereale (rye), Salix spp. (willow), Eucalyptus spp. (eucalyptus), Triticosecale spp. (triticum- 25 wheat X rye), Carthamus tinctorius (safflower), Jatropha curcas (Jatropha), Ricinus communis (castor), Elaeis guineensis (oil palm), Phoenix dactylifera (date palm), Archontophoenix cunninghamiana (king palm), Syagrus romanzoffiana (queen palm), Linum usitatissimum (flax), Brassica juncea, Manihot esculenta (cassaya), Lycopersicon esculentum (tomato), Lactuca saliva (lettuce), Musa paradisiaca (banana), Solanum tuberosum (potato), Brassica oleracea (broccoli, cauliflower, brussel sprouts), Camellia sinensis (tea), Fragaria ananassa (strawberry), Theobroma cacao (cocoa), Coffea arabica (coffee), Vitis vinifera (grape), Ananas comosus (pineapple), Capsicum annum (hot & sweet pepper), Allium cepa (onion), Cucumis melo (melon), Cucumis sativus (cucumber), Cucurbita maxima (squash), Cucurbita moschata (squash), Spinacea oleracea (spinach), Citrullus lanatus (watermelon), Abelmoschus esculentus (okra), Solanum melongena (eggplant), Papaver somniferum (opium poppy), Papaver orientale, Taxus baccata, Taxus brevifolia, Artemisia annua, Cannabis saliva, Camptotheca acuminate, Catharanthus roseus, Vinca rosea, Cinchona officinalis, Coichicum autumnale, Veratrum californica, Digitalis lanata, Digitalis purpurea, Dioscorea spp., Andrographis paniculata, Atropa belladonna, Datura stomonium, Berberis spp., Cephalotaxus spp., Ephedra sinica, Ephedra spp., Erythroxylum coca, Galanthus wornorii, Scopolia spp., Lycopodium serratum (Huperzia serrata), Lycopodium spp., Rauwolfia serpentina, Rauwolfia spp., Sanguinaria canadensis, Hyoscyamus spp., Calendula officinalis, Chrysanthemum parthenium, Coleus forskohlii, Tanacetum parthenium, Parthenium argentatum (guayule), Hevea spp. (rubber), Mentha spicata (mint), Mentha piperita (mint), Bixa orellana, Alstroemeria spp., Rosa spp. (rose), Dianthus caryophyllus (carnation), Petunia spp. (petunia), Poinsettia pulcherrima (poinsettia), Nicotiana tabacum (tobacco), Lupinus albus (lupin), Uniola paniculata (oats), Hordeum vulgare (barley), and Lolium spp. (rye).

[0215] In some examples, a monocotyledonous plant may be used. Monocotyledonous plants belong to the orders of the Alismatales, Arales, Arecales, Bromeliales, Commelinales, Cyclanthales, Cyperales, Eriocaulales, Hydrocharitales, Juncales, Lilliales, Najadales, Orchidales, Pandanales, Poales, Restionales, Triuridales, Typhales, and Zingiberales. For Attorney Docket No. PIV-00053 WO

[0216] example, the methods described herein can be performed on plant(s) belonging to the class of Gymnospermae are Cycadales, Ginkgoales, Gnetales, and Pinales. In some examples, the monocotyledonous plant can be selected from the group consisting of a maize, rice, wheat, barley, and sugarcane.

[0217] In some examples, a dicotyledonous plant may be used, including those belonging to the orders of the Aristochiales, Asterales, Batales, Campanulales, Capparales, Caryophyllales, Casuarinales, Celastrales, Cornales, Diapensales, Dilleniales, Dipsacales, Ebenales, Ericales, Eucomiales, Euphorbiales, Fabales, Fagales, Gentianales, Geraniales, Haloragales, Hamamelidales, Middles, Juglandales, Lamiales, Laurales, Lecythidales, Leitneriales, Magniolales, Malvales, Myricales, Myrtales, Nymphaeales, Papeverales, Piperales, Plantaginales, Plumb aginales, Podostemales, Polemoniales, Polygalales, Polygonales, Primulales, Proteales, Rafflesiales, Ranunculales, Rhamnales, Rosales, Rubiales, Salicales, Santales, Sapindales, Sarraceniaceae, Scrophulariales, Theales, Trochodendrales, Umbellales, Urticales, and Violates.

[0218] Other non-limiting examples of suitable plants include mosses, lichens, and algae. In some cases, the methods described herein are suitable for any of a variety of transgenic plants, non-transgenic plants, and hybrid plants thereof.

[0219] Definitions

[0220] The terms "polynucleotide," "nucleotide sequence," "nucleic acid" and "oligonucleotide" are used interchangeably. They refer to a polymeric form of nucleotides of any length, either deoxyribonucleotides or ribonucleotides, or analogs thereof.

[0221] Polynucleotides can have any three-dimensional structure, and can perform any function, known or unknown. The following are non-limiting examples of polynucleotides: coding or non-coding regions of a gene or gene fragment, loci (locus) defined from linkage analysis, exons, introns, messenger RNA (mRNA), transfer RNA (tRNA), ribosomal RNA (rRNA), short interfering RNA (siRNA), short-hairpin RNA (shRNA), micro-RNA (miRNA), ribozymes, cDNA, recombinant polynucleotides, branched polynucleotides, plasmids, vectors, isolated DNAof any sequence, isolated RNA of any sequence, nucleic acid probes, and primers. A polynucleotide can comprise one or more modified nucleotides, such as methylated nucleotides and nucleotide analogs. If present, modifications to the nucleotide structure can be imparted before or after assembly of the polymer. The sequence of nucleotides can be Attorney Docket No. PIV-00053 WO

[0222] interrupted by non-nucleotide components. A polynucleotide can be further modified after polymerization, such as by conjugation with a labeling component.

[0223] The terms "polypeptide," "peptide" and "protein" are used interchangeably herein to refer to polymers of amino acids of any length. The polymer can be linear or branched, it can comprise modified amino acids, and it can be interrupted by non-amino acids. The terms also encompass an amino acid polymer that has been modified; for example, disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation, such as conjugation with a labeling component.

[0224] As used herein the term "amino acid" includes natural (e.g., alpha-amino acids) and unnatural or synthetic amino acids, including both the D or L optical isomers, amino acid analogs, and peptidomimetics. Amino acids can be positively charged or negatively charged. Amino acids can be not positively charged, or not negatively charged. Amino acids can contain or lack sulfur.

[0225] As used herein, the term "about" is used synonymously with the term "approximately." Illustratively, the use of the term "about" with regard to an amount indicates that values slightly outside the cited values, e.g., plus or minus 0.1% to 10%. As used herein the terms "microorganism" or "microbe" should be taken broadly. These terms, used interchangeably, include but are not limited to, the two prokaryotic domains, Bacteria and Archaea. The term may also encompass eukaryotic fungi.

[0226] As used herein, in the context of non-intergeneric microorganisms, the term "gene- edited" is used synonymously with the term "engineered." An engineered microorganism contains in its genome at least one genetic modification. In some embodiments, the bacteria of the present disclosure have been modified such that they are not naturally occurring bacteria.

[0227] As used herein, when the disclosure discusses a particular microbial deposit by accession number, it is understood that the disclosure also contemplates a microbial strain having all of the identifying characteristics of said deposited microbe and / or a mutant thereof.

[0228] The term "microbial consortia" or "microbial consortium" refers to a subset of a microbial community of individual microbial species, or strains of a species, which can be described as carrying out a common function, or can be described as participating in, or leading to, or correlating with, a recognizable parameter, such as a phenotypic trait of interest. Attorney Docket No. PIV-00053 WO

[0229] The term "microbial community" means a group of microbes comprising two or more species or strains. Unlike microbial consortia, a microbial community does not have to be carrying out a common function, or does not have to be participating in, or leading to, or correlating with, a recognizable parameter, such as a phenotypic trait of interest.

[0230] As used herein, "isolate," "isolated," "isolated microbe," and like terms, are intended to mean that the one or more microorganisms has been separated from at least one of the materials with which it is associated in a particular environment (for example soil, water, plant tissue, etc.). Thus, an "isolated microbe" does not exist in its naturally-occurring environment; rather, it is through the various techniques described herein that the microbe has been removed from its natural setting and placed into a non-naturally occurring state of existence. Thus, the isolated strain or isolated microbe may exist as, for example, a biologically pure culture, or as spores (or other forms of the strain). In some embodiments, the isolated microbe may be in association with an acceptable carrier, which may be an agriculturally acceptable carrier.

[0231] In some embodiments, the isolated microbes exist as "isolated and biologically pure cultures." It will be appreciated by one of skill in the art that an isolated and biologically pure culture of a particular microbe, denotes that said culture is substantially free of other living organisms and contains only the individual microbe in question. The culture can contain varying concentrations of said microbe. The present disclosure notes that isolated and biologically pure microbes often "necessarily differ from less pure or impure materials." In general, the term "genetic modification" refers to any change introduced into a polynucleotide sequence relative to a reference polynucleotide, such as a reference genome or portion thereof, or reference gene or portion thereof. A genetic modification may be referred to as a "mutation," and a sequence or organism comprising a genetic modification may be referred to as a "genetic variant," "mutant," or "engineered."

[0232] Genetic modifications introduced into microbes can be classified as transgenic, cisgenic, intragenomic, intrageneric, intergeneric, synthetic, evolved, rearranged, orSNPs.

[0233] 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 disclosure pertains. Methods and materials are described herein for use in the present disclosure; other, suitable methods and materials known in the art can also be used. The materials, methods, and examples are illustrative only and not intended to be limiting. Attorney Docket No. PIV-00053 WO

[0234] The invention will be further described in the following examples, which do not limit the scope of the invention described in the claims,

[0235] EXAMPLES

[0236] Example 1: Design of strains and plasmids for bacterial genome modification

[0237] The following strains (Table 4) and plasmids (Table 5) were prepared to determine the effect of the prophage grounding on host cell growth. The ammonium secretion mutations disclosed in Table 4 were described, for example, in WO2019084342A1, assigned to Pivot Bio, which is incorporated by reference herein.

[0238] Table 4: Host Bacterial Strains

[0239] Strains Species Genotype Referred to as...

[0240] ST18 Escherichia coli S17 Xpir bhemA Mating strain 5812 Kosakonia WT wildtype oryzendophytica

[0241] 5812- Kosakonia Phage_A3prime-2 3' prophage 7663 oryzendophytica deletion; attR deletion 5812- Kosakonia kxerD Prophage 7767 oryzendophytica recombinase deletion 5812- Kosakonia AnifL_PompX_y2-nifA; glnD- Grounded (5 + 8686 oryzendophytica AUTase (Ammonium Secretion mCherryV2 Mutations), bxerD, tsxerC2,

[0242] Ev<erC3, pB61A Plil-mCherry- ParABCDE_V2

[0243] 5812- Kosakonia AnifL_PompX_v2-nifA; glnD- Grounded CD + GFP 8687 oryzendophytica AUTase (Ammonium Secretion

[0244] Mutations), HorerD, tsxerC2,

[0245] &xerC3, pB61A Plil-GFP- ParABCD

[0246] 5812- Kosakonia AnifL_PompX_v2-nifA; glnD- (D Knock out (KO) + 8683 oryzendophytica AUTase (Ammonium Secretion mCherryV2 Mutations), Phage_A3prime- 8, pB61A_Piil-mCherry- ParABCDE_V2

[0247] 5812- Kosakonia AnifL_PompX_v2-nifA; glnD- CD Knock out (KO) + 8684 oryzendophytica AUTase (Ammonium Secretion GFP Mutations), Phage_A3prime-

[0248]

[0249] 8, Pb61A_Plil-GFP-ParABCD Attorney Docket No. PIV-00053 WO

[0250] 5812- Kosakonia AnifL_PompX_v2-nifA; glnD- Nitrogen Secretion 7003 oryzendophytica AUTase (Ammonium Secretion Mutant Mutations)

[0251] 5812- Kosakonia AnifL_PompX_v2-nifA; glnD- Nitrogen Secretion 8252 oryzendophytica AUTase (Ammonium Secretion Mutant with Mutations), IXxerD, EsxerC2, Grounded < D

[0252]

[0253] &xerC3

[0254] Table 5: Plasmids

[0255] Plasmid Name Plasmid Features

[0256] pB61A Plil-mCherry-ParABCDE. V2 Kanr, Rk2 origin, oriT

[0257] pB61A Plil-GFP-ParABCDE Kanr, Rk2 origin, oriT

[0258]

[0259] Example 2: Bacterial gene modification methods

[0260] E. coli strains carrying conjugation plasmid pB61A plasmids were grown aerobically at 37°C in super optimal broth (SOB) supplemented with 5-aminolevulinic acid (50 pg / mL) and kanamycin (50 pg / mL). K. oryzendophytica strains were grown aerobically at 30°C in SOB and K. oryzendophytica strains harboring conjugation plasmid pB61A were grown at 30°C in SOB supplemented with kanamycin (50 pg / mL).

[0261] Genome modifications were made in strains by two-step allelic exchange (Hmelo et al., 2015). Briefly, integrative plasmids were constructed using the Gibson assembly (Gibson et al., 2009) recombinant DNA technique (New England BioLabs). Gene deletions were carried out using two-step allelic exchange of the native open reading frame (ORF) with the truncated ORF. The deletion constructs were sequenced by Sequetech (Mountain View, California).

[0262] Plasmids harboring fluorescent proteins were mated into K. oryzendophytica using an E. coli ST18 strain harboring the conjugation plasmid pB61A.

[0263] Example 3: DNA sequencing procedure

[0264] For initial whole genome sequencing (WGS), 1 ml cultures of stationary culture were pelleted and frozen prior to genomic DNA (gDNA) extraction. For prophage triggering experiments, 100 pL of cultures were aliquoted into tubes and frozen until they were used for gDNA extraction. gDNA extractions were performed using the QIAgen Blood and Tissue kit (QIAGEN GmbH, Hilden, DE), using the bacterial methods specified by the kit. The gDNA Attorney Docket No. PIV-00053 WO

[0265] was synthesized into Illumina-capable sequencing libraries using the Illumina DNA Prep kit. Libraries were pooled and sequenced on an Illumina MiSeq, NextSeq 1000 sequencer.

[0266] Example 4: Bioinformatics for prophage region prediction

[0267] Genome sequences were assembled using Unicycler Hybrid Assembly (Wick et al., 2017) and utilized for downstream analyses. Prophages and integrase genes were predicted from assembled genomes using PhiSpy (Akhter et al., 2012) and VirSorter2 (Guo et al., 2021). The genes within predicted prophage regions were annotated using DRAM (Shaffer et al., 2020). To detect prophage activation, sequencing reads were mapped to the assembled genome using Bowtie2 (Langmead et al., 2009). Read depth was then determined in predicted prophage regions using the SAMtools mpileup function (Li, 2011). The resulting files were examined for increases in coverage within predicted prophage regions.

[0268] Example 5: Induction of prophage using mitomycin

[0269] Bacterial strains were triggered for prophage activation by treatment with mitomycin (2 pg / mL) in either stationary phase (after at least 12 hours of growth in liquid) or during exponential growth (after a 20 pL back dilution of cells from a stationary culture into 1 mL of fresh media and growth for 4-6 hours). After mitomycin addition, cultures were returned to the incubator for overnight incubation (at least 12 hours of growth). Whole genome sequencing was performed. Samples were analyzed for the presence of activated prophages by determining if there was increased sequence read coverage in whole genome sequencing reads over predicted prophage regions.

[0270] Example 6: Identification of an active prophage in a bacterial host ceil

[0271] While growing the K. oryzendophytica strain 5812, bacteriophage plaques were observed. DNA sequencing showed an increase in sequence read coverage of the bacterial genome in the area that encoded the prophage P-5812-01, confirming that the P-5812-01 phage was activated during growth. Prophage P-5812-01 was identified as being located downstream of the folD gene, a bifunctional methylenetetrahydrofolate dehydrogenase also referred to as the ads gene (exemplary sequence is described by Accession No. Attorney Docket No. PIV-00053 WO

[0272] MGMEDFLC_02997) and upstream of the hypothetical protein encoded by the gene having Accession No. MGMEDFLC_03078.

[0273] The bacterial strain was tested for prophage activation using mitomycin, following the method described above in Example 5. Sequencing read coverage analysis was performed on two replicates of the bacterial strain K. oryzendophytica strain 5812 and showed the induction of an active prophage after induction by mitomycin (FIG. 6A). FIG. 6B and FIG. 6C show the complete deletion of the recombinase (strain 5812-7767) and the complete deletion of the 3' attR region (strain 5812-7663). An increased sequencing read coverage was observed for both strain replicates, signifying consistent replication of the prophage after exposure to mitomycin, and positively identified its location in the genome of K. oryzendophytica strain 5812. Subsequently, two additional prophages were identified by increased bioinformatics.

[0274] Example 7 Designing a grounded prophage.

[0275] Using the methods described in Example 6, an active prophage (P-5812-01) was identified in K. oryzendophytica strain 5812. The phage DNA sequence was identified (SEQ ID NO: 12) following the method described above in Example 3 and Example 4. The regions corresponding to the attl, attR, and the phage recombinase were identified. FIG. 3 is a general map of the phage sequence identifying the areas that could be deactivated or deleted to generate a grounded prophage.

[0276] Example 8: Bacterial genetic modification to disable prophage activation

[0277] Phage grounding: Once identified in Kosakonia oryzendophytica strain 5812, grounding of the prophage (P-5812-01) was completed with the construction of a recombinase deletion (strain 5812-7767, deletion of xerD gene encoding XerD recombinase), and a 5' deletion (strain 5812-7663). Gene editing was performed as described above in Example 2. The resulting "grounded" or modified prophage sequence is shown in SEQ ID NO: 12. The resulting strains are shown in Table 6, where "+" indicates that the strain contains an active genetic element, and indicates that the indicated genetic element was deleted.

[0278] Table 6. Engineered Strains

[0279] Strain Recombinase (XerD) attR

[0280]

[0281] Attorney Docket No. PIV-00053 WO

[0282] 5812

[0283] 5812-7657

[0284] 5812-7767

[0285] 5812-7663

[0286]

[0287] In Kosakonia oryzendophytica strain 5812, an additional two prophages were identified via increased bioinformatics and were identified as prophage 2 and prophage 3. The two putative recombinases from those prophages (XerC2 from P-5812-02 and XerC3 from P-5812-03) were also deleted. The resulting modified prophage sequence of P-5812-02 is shown in SEQ ID NO: 13 and was located downstream of the mug gene (Accession No. MGMEDFLC_03290) and upstream of the hypothetical protein encoded by the gene having Accession No. MGMEDFLC_03335. The resulting modified prophage sequence of P-5812-03 is shown in SEQ ID NO: 14 and was located downstream of the hypothetical protein encoded by the gene having Accession No. MGMEDFLC_03659 and upstream of the rplS gene (Accession No. MGMEDFLC_03688). The resulting recombinase deletion strain was tested for prophage activation and showed a drastic decrease in read coverage in comparison to the WT prophage (FIG. 3C). This confirms the successful grounding of the prophage by the process described herein.

[0288] Trimming the 5' end of prophage P-5812-01 would have resulted in a recombinase deletion and also deletion of the attL DNA sequence, because the recombinase gene is adjacent to the 5' end of the prophage. Trimming the 3' edge of the prophage was accomplished via a kb deletion overlapping the 3' end of the prophage and the beginning of the bacterial host sequence, as determined by increased read coverage of the prophage region. The resulting 3' trimmed grounded prophage strain was tested for prophage activation and showed a drastic decrease in read coverage in comparison to the WT prophage (FIGs. 6A-6C). This confirms the successful grounding of the prophage by the process described herein.

[0289] Prophage P-5812-01 (SEQ ID NO: 12) TCTAAGCCGTAGGTCACAGGTTCGAACCCTGTAGGGCGTACCATTTAAAATCAAATGCTTACGCAAG TCTCAATCCAGCCTGATTTCCTCCTTGTGTCGTATTTGTGTCATGGTTGCCAAAAATGGCATCAATTTT CCGCGCGTGCTCACTCAGGTGATTAGGCGCGAGGTGCGCATAACGTCTCACCATCTCTATTGACTCCC AGCCTCCCATTTCCTGCAATACGGACAGCGGAACGCCGGACTGAATCAGCCAACTCGCCCAGGTGTG CCGGAGGTCATGAAAACGGAAGTCTTCGATGCCTGCTTTCTTCAGGCCAATGTTCCATGCCTGATTAT CATCAACACGCATTTTCCTGACTGCTGGTGTCAGCGTGCCGTCTGGCCGGTGCTTTGCCTTTGTATGC Attorney Docket No. PIV-00053 WO

[0290] ACAAATACCCACCGTGAGCTTTTACCTATCTGATCCCTCAGAACCCTGCATGCGGTATCATTCAGAGC GACGCCGATAGCCTTGCCCGCTTTTGCGTTCTCCGGGTTTACCCATGCAACCTTTCTCTGCATATCGAC CTGCTGCCACTCCAGATCAATGATGTTGGAGCGGCGCAGGCCGGTAGCCAGTGCAAAAATCACCACC GGCTTTATGCTTTCCGGCATACACTCAATCAGCCTTTCGGCCTCATCGCGAGTCAGCCAGCGAATGCG CTTACTGACCGGTTTCTTTGTTTTAATTACCGGGGCCGCCTTTATCCACCCCCAGTCATTGGCCGCCGC TCGAAGCAGGGATCGCATGAAAGAAAGATGCTGGCTTTTAGTTGCCTGGCTAACCGGCTTTGCCACA TATGCTGGTGGCTCTTGACCCTTCCGCAATGCGGAATCTCGCCGCGATTCCCAGACCTGCAAATGCTT ACGGTTAATCATTTTCGACACAGAATCATGAACCTGTTCGGCTGTAATGGTCGAAATATCCCGTCCTG AGAAATGTTGCAGGAAAAATTCTATCTTCGTCCTGTCATCATCCAGAGATCGCTTATCATCTTTCTCCC GCAACCAGCGGATGCAACACTCTTCAAACGTCCTTGTTGGCAAATCACCTATCTGATCCACCCGCCAC GCTTCTGCCTTCAGCTTGTCGTGCAATTCCTGCGCTTGCTTTTTGTCCCCCGTACCAAGAGATCGTCTA ATTCTTTTCCCTGACGGCGTAACGAAATGACAGTGCCAGACGCCGCCCCTGAGGGTGATTGACATAA AATTTCTCCTTTATGTTCACCCGCGCTCGCCGTAACAGGATCGCGCCGGGCGTGTAAATACGCAATAC AGGCCGCATCGGTCGTGCGGTATTTGTTCCCGATCTTCTGCCCGGCCAACTGACCAGAATCAATCAGT CTGTAAATAGTGCGCGGAGACACGATCAGGAATTCCGCCGCCTGCTGCGCGGTGATTGGCTTTTCTG AAACCATTGGTTTACTCCAAGCAATAAAAAGACCGCTCATGGCGGCCGCATTAAACCAGTTTCTTGTG CATCATCGGGCGCTCAGTCGTGGCCCACATCAGATCCTTCTCTGCATGCGATGTGAAGTCACCAGAG TCCCATTCCACGCGATACCACGTAGGCCTGCCTTCCTGTTCAATAAATCCCTCCACCACACCCCGAATC CCTCCGCTTACATGCTTCACAATTGCACCAACCGGGAACTTAGCCATGAATGCCTCGCATGATGAATA AGCCAAAAAGAAGCCGCCCGCAGGCGGCAAACAGGGAATTAACATTGGTCTTATCGGTGCTCAGCA CCCAACAGGCGACTCAGTGAATCGCCTGTAAGCTGCCCTAATCGTCTTCGTCATCATCCCATTCATCCC AGTCTTCATCATCCTGAGAAAACGATAGCAGGGGGTTTGTGGCCGCCAGCATTTCACTGGCAGCGCC ACGGCGCTGAAGCCTGCGCAGTGCTTCATAAAGCTCGAACGCTTCGGTGCGTTCATCGCCGATATCG AGAGAGCATGCAACCCGGTGCGCGGCTGTGACGATGGTTTCTAACTGGTTGCGAATATCCTGCATAG TTTCCATATCACATCCCCCTTTGCTTACGCTTAATCTCGATAATTTGCTGGCAATCCGCGCATGTCTGG CAGCCGGGCACCGCAACCCGCCGCGCTTCTGGGATATCCTCCCCGCATTCCTCACAGTATTCCGCTGA TACCGCGTTACGGTTGATACGGTGCGCCTGAATGGCATTGTTGATGTTCTGCTCTACAAGCTCGTTGG CTTGGTCGATTATTTCTGCGCTCATGCTCTGATCTCTCCCGGCAAAGGCTTCATTCCATCTGTTAATGA CCGCTCAATGTGGCTTAGAACCGTGCGCAGTCTTTTAACCTCTrCATCGCTGGCTAGTCGCCGGGTTG CTTCCCATAGCGTGCGAGTTAGCACATAAATTGGAACATCGCGCTCAAGCCCTAATGCCTGAGCAAG GCCTCGCTGCGCATCATTAAGGCTTTCCTCAGCCTCTTCACGACGCTTACGGCGGCGGGCAAGCTCCT CTTCTCCTTCCTGGCGAATCTGTTTCATCCGTTGTTCTTGAGCGTTGATCTCTTGTTCAAGCCGGTCTrT CGACCTGTAGGCGCGACATACCAGATCGGCAATTTCCGCCCCGTGTTTCTTCAGCAATTTATGATCAA TAAAATACGCGTGCGCGGTGCGCGATTTAATCTGTGTGCGCTCTGCTTCCCGCGTCAGGCCATCCATC AGCAGTTTCATCCAGGCATCGCGCGGTAAGTTATCAAGCTGTCGAATGGTTGGGCCTTTCAGCGTGT GCCAGCCTGATTCTTTACGGATCATCAGGCCGCAGCCTTCCGGGATGTCGCCTTTCTTCAACAATCCT TCTGGTACCGCAAACACAACACCGCCAGCGTATTTGAAATACTTGGTGTACTTCCCGGCGGTAACGTC TGCCCGAAAGTCACTAACGCTAACTTTCACCTCGTATACGACCGGGCAAAACTTGCTGTAGCTGTGA GCCATTGCGTAAACATCAGGGCGACATGTCCCTGCTGGCCCGAGTTGCATATCCTCCCAGCAAATGC GTGCGGTATTGCGCCGCAGGTGCTCGGCGAGGTCGTGAGCAAGCTCGTTATGGCCCCATTTCTTTTC GATCATTGAGTTGCTCCTCTGAAAGTAAGAAATGTACCCATCGCTTTATCGACCAGCTTTGAGTTGTG GTATTTGGCGATAGACCAGGTGATAGCGAATAAAATCCACCGGAAATGGCTGGTGTAAGTCTTAAA GGTCAGGCCGTCACAGATATCCCACCTATCGAATCTTTCAGGCCAGTCAGCTTCATAAACCGCGTTAT AGGCTTCATAATCGTTACTGAATCCAGCACTGCACAATTCACGAACGATTTCCCGCACCCTCGCTTTAT CGCTATCTGGCGTTTCGTCATCGTCATCCCAGTCTTCGTCGTCTTCTTCATCCTCTTCAGAATCCTCAAG GTAATCGCTCAGGGATTCTTTCAGGCTCAGGCAGAAAGCATCGTGGTCGTATTCCTTCGCCAGCAGT TCACGCGCAGAGCAACCCGCGCCAGCTTCCAGTTTTTCAGACCAATAGTCGGTATTAATCCCGCCTTC CCAGGCCCCGAAAAAGTTGAACATGTCCGCAATGCGGGAAAACGTCCACGTTCCCATGTCGCCGGTA ACGGTCAGGTAACCCGGCCAGGTGATCACATCGTAGTAATAGCAGGAGGTGCCAGGCTGCTGCATG CGCAGATGGCGATAAAGCCCATCATCACGGATAATTTCCAGATGATGGAATGCAGTATCCAGAAGA Attorney Docket No. PIV-00053 WO

[0291] AAGCGACTTGATGTGTCAAATTGACGGCGATTCACGATTCCACTCCATAGCGCCCATTCATGCGCCCA ATGCTGCTAACGAAAGCCACAAGGCTGACACCCATAGGCTCTATTTTTTTGTGGTGCTTTTTGAGGAT CGGCGGCACCGCGTCATTCCATTTCGGCTTAGGCCGGGCTTTCATGGCCTGGCGGATTTCGTCGACG CATTTACGGCCCTGCGCGCGGATGGCGTTTTCTTTCTCTGCTGGTGTCATGCTGCCTCCCGGCGGGCG ATGAGTTTTGCCCCGAAAGTCATTAACTCGTCCCGATCCACAGTTGCGAAGTGGCAGTGTGTACGCG GGTACGGATGCCAGATAATGAGCATCGAACCTTTGTTATTACCGCTGACTGGCTTTCCGGTTACCGG ATTGATGAATGCCAGCCGGCCGCCAGTGATGAAGCGAACCTCACTTGCGGTCTGGATCGCCTCTTTA AACCAGCCACAGACGTATCAGATGGAACCAGCATTACGCAGCCAATGCAGCCGCAGCGGTTTTGATC AGCTGCTTTGAGGACGAATGGCGTTATGTCGCTATACGGAGGATTGAGCCACACATAGCCGTATGCA TAGGCCATTGCTTCAGCCCACGAAGTGGAGAGTGTGTTTTCTTCCTCTGTAATGAACAGACGACACA GACGGTTTTCTTCAGTGGCCGCCGCATCTAACTGGAAGATAAATTCCGCATTCAGCGCTGCGAAGAT TTCCGGCGGTGTTCGCCAGCTATCACGATGCTCTGGTGGGGTAGTGCTTCCGCCGTAGTCGCTCATG CTTCACCGCCTTTGACAAAGATrATCCAGTGCGTTTTGTCGGCTTTGCCGGTGCGCTGCCAGATGGCT GGCTTCTCGTCAGTCAGCGCCAGAATCTGGCTCACCGGGATTTGCGTTTCGTTCCATTTGAAGATGAG CACGCCGTGTGGCCACAGCACGCGGAATGCCTCTTTGAAACCTGCACGCAGGTCATCGCGCCAGGTG TCTTTGTTCAGGCGTCCGTATTTTTTACCCATCCAGGCGTTTTCGCCGACCCGCTCAAGGTGTGGAGG GTCAAACACAACAACAGGGAAAGATGAGTCACCAAAAGGCAGCGCGCGGAAGTCGGCGATCACAT CCGGGCTGATAACGAGACTGCGACCGTCGCAAAGCGTATGCTGCTCGGCGCGGATATCGCTGAAGA CTGCGCGTTCGTCCTGCTTATCGAGCCAGAACATACGGCTTCCGCAGCACATATCGAGAATGCTGAA ATCACTCATGACTCACGCTCCGGGTCGAATTCAGGCCAGTTATTGCGCTCGTAGTTGGCTTGTACCCG CCGCGCCTCAACTTCCTCAATGCTGCGCCCGGTGATTTCAGCAACCTCTGTGTTGTTGTGTCGCCAGA GCAGGGCAAGCTCTTCAGGTGTCCAGCCGGACATAATTTTTCCTCCTGTTACCCGTGCCAGTGAGTTG CGTTTACTTCGGACGGAGTGAATAGAGCGGCCAGTGGCCACTGCGATTTCTTCCTCGGTGAAGCGAC CAAAAAGAAACAACTCCGCTTTAGTCCACGGTCTGCCGATCATTCGTGAGGGGAGGTTGGCGCCAAT CCTTGACGCCTGCCTGGTGATTGCCGCTTCGGAGCGTTCGAGCTTCGCGGCAATCTCTTTCAGTGGCA TTTTTTCGCAGACATCATGCAGGAAGAGATTTTCAAAGGGCTGCCAGCGTTCACACCGCATAGCTACC TCCATTGTTCGCCGAACGTAAATCCTATCTCCGCGAGCGATTCATCCATCTTTTCGATGAACTCAGGC ACCATTTCGTCGAAGTCAGCCATAAATTTCGGATCACGTTCGACGACTACGTGATGAATGCCTTCTCG CTTCATGCGCGGGTCATAGTTGGCAAAGAACCAGGCGTTCTTTCCGGTAACCCACATGCTGTACTGA ACCTGCGCCATATAGGCCGATTTGATAGCCTCAAATCCACCCAGGCGGAACTTCATGAAGTCGCGGG AAGTGAACGGGCATTTCAGCTCAAGGCCGAAATCGTTACTGCACAGCCCATCAGGTGAGCATGCAGT GCGCATGGTTTCATCGCGAAAGAGGATCGGCGACTCTGTAACGTTAACGCCGGTGGTGAATTCGAA GAGCGTGCGCGCGTCCTCTTCATACTGTTTTCCCCATGCTAGCGCCTTGGCGTTAACCTCTGGCGCTA CGCCGGTGCAAACTTCAGCGAGCAGGGTGTGGAAGTAGGACATCTTCATGTCCGTCCATTTCGTACC GGATCGAGGTTTGGAAATGACGTTGTGCACTTCAGACGCAGTAATGACGCCAAGGCGCAGCTTGTG CCACGCCTCATCGCCTTGTTGGATGGTTGCTACGTCAATACCGGTACGCGCGAGGATGATTTCTGGT GTCATGCTGCTGCCTTCTGCCTTAGGAAACCGAGAGCCTTAACGCCTTCCGGTTCTGTCAGGTCGGCG GGCTGAGAAATAGTGCGTTTGAAAATGCGCGAGCAGAGAGGGAGAAGATCTGCATCCCATGTCTTA TCCATCGAGACCAGCAGGTCGTTAATCTCTTTTTGCGTGGTTTCGCTAAGCGGCGTTATATCGCGCTC TGACTGACGTTCTGCTGCGAAGTTGATACCTTCTTCGCCCTCGGTGTTAACGTGGTCGATAGCGGCAT CCAGGCGCTCGCGGCGAGGCCAGTATTTGGCAGCCTGCTTAACCACCGTCTTGAGGATCATTTGCTC CTCATCGGTAACCCACGGGCATTnTrACTGTTGTCAGATTTATATTTCTTCCATGCTTCTGACCGGTC ACGGATGGAGTAGATGGCATCAATGCGCATCGTATGGGTTAGGTAATCGCCATCGTCAGTTTTTACC GTTACATACGCGCCTACGATCTCCCCGCGTTGCTCTTCGGTATCAAAGTCGTTGTAGATGTGGATCGG TGGCTTATCGAGCCCTTCGCGGCGGAACTGGTCGTTTCTGCGAACAATTGCCGACTGACACCACTTG ATTGCTCCAGACTGCTGTGCAATGTGCATCAGGCCCATGTAACTGATATCAAGGCAAATGGCCCCTTT GCGCGGAACCAGGTAAGCCAGCTTCTGCGCAGGGTTTAGCGAAATGCCGATAGCCGCAACGTTGAC GATAGCATTCTGCGTGCTGGTCTGGTTCTGGAATGCAACTTTTGCCAGGTAATCGTTATTCTGAAATA GCTGGATGGCGAACTGGCTTTCCTTCGCCCACACCATCCGCTCGTCAGTGGCCGCCTTAATGAAAAG CGGCTCTTGTTGTTTGACAAAATCAATAAGGGTTAAGCTCATAATCCCTCCTTAAAACGGGCAGCCGG Attorney Docket No. PIV-00053 WO

[0292] TGCGGTGTTCCCATTCGTAATCAGCCTGGGCGTAAGCAACAGCGGTTACGAAGTCGTTGTAGGCTTC GGTGGCGTTATCATGGCTGAGTCCGGCATAAGGTTTTGTTTCAACCGGGATAGAGAGAAACACCATT TCGGCAGGCGTTTTAGGGAAGATGGCGATAATTTCrTTCGCCCGGTCATCAACCCATTTTTCACGCTC AATTTCCGCTTGGTGTTGGAAAGCGCGTCGCGCCTCGATACGGTCGTAAGTCAGGTAGGCGTTCATG GTCACTCCTGAAATTTGGTTGTGCGGATCCGGCCCGCGTTTAGCCAGGCAGATCGGTTGAAAAGGG AAGTGGTTACTTATTGGTCAAAGCAGATGCAGGGACTTCATCAGCCCGAACAATTCGCTCTACCGGG AAGCACTCACCGGCAATGCGCTGCTCGACAGCCGCGTCCTCGCATAGCGCATACGTTTCATAAACGC CGATAACGACATCCTGAAATTCACCGTTTGTCATGCCAACGGTCAGGACTAACGCGAATAACGTTCCC ATCAATGAAGCTCCCTCCCGGTTGATTGCCGGAACTGATCGGCGGCCTGGTTCCACATCGCCTCGTTA CCGAGGTATTCGGCGATAGCCGCTTTGCTTTGAGCGGCGCGAGCCTGGTTAATGTCGACCGTTTCTG TTAGCTCCACGATGTGCCTGAATCCGGCGAGGATCATTTGCTGGCGGTTTAACGTGAAACCCTGACG GGGTTTATCGACTTGGGTAAGTCGCCATTCGTAGCCGTTAGTCAGCTTGTGAACTGAGTACTGAATG CCTTTGTGTGTGACGATTTGCTGCATGGTTTCCTCTTCGCCTTATCGCCGGCCAGCGGAGCTATTAAC CTTTGCGCATAAAAAAAGGCGCTGGATGGGCGCCTGTGTACTGGATAACGAAATGCCCTCGTAAGG GCGCTTGGGTATCCAATAAAAAACCCGCCGGAGCGGGTTAGTCGTTTGTCACTGTGTAGCCTTCGTT TTCAAGCCAACTAACAACCTCAGACTCATCCATGACGTCAAGTAGTGCGCTTGCGGTGTACTCAGAA ACGATCTCTTCAGGCTTTACGGAATCAACAAGTTCAATGCCGTCAATGTCGATTTCCATGTGCCGGTT ATATCCGTCGCCGTATGGGTTAATTGAGTTGATTTGTTTTACTTTTAACGTTGCATCAATTGCCATCGC CTTACCCTCTGTAGTTACCCGCTGATGCGGTTATGTCTCATCGACGCATGGAAGAGCTTTCCAGTCGA TATCGTTCTTTGCCGAATCCCACAGTCCTATATTTTTGTACTTTTCTGAGCATGGAATGCAGATGTTGT AAGGCCTGCCAACCATGTTTAGAGATATTCTCTGCATCTTTTCAGATGCCAGGTAGTACCCACAAAAT TGACACTTGTGCATACGTGTTCTCCATTTAGTTACCCGCTGATGCGGGAGAAATGCTTTGGCGATTGG ATGGCAGGCGCTGACTTCTTCCTGCTTTCTGGATGGCACTAAACCAGACCTTGTACGGCATATGCCGC GTATACCATCTATCACTGCGCATAGCGGCTTACGTTGCGATCTCGGTGCGCATCAGCCTGCGCATTCA TCCAATCCCAAAACATTCCCTGTATTGGTCAGCGCCAACTTCCTGCCAGTGTTGCCCGTTCTCACGCCG TTCTCGCTCTCGCGCGGGGATACTCTCTCACCGACCGGATCGCACCCGGTGATACAGCACGTTTTCGT GTAAGGGTCTTAACAGGTCATTGACGCTGTAAATCTGCATGTTGTTAAAAAGCAGGCTGACTCATGT CTGCCGCGGCTTAACTTCCGGTGGCCGCATCGCTGTGTTGTCGCGATGGGTTAAAGATACAGATAAA ACTGTATTTCCGTCAACAGACAAAACTGTAATTTGCGTTGTAATTTACACATGTATCTGTAATAGAAG GTTATTTATTTTTGTTGAGGCGAAAAAAACCGGCAAATGCCGGTTTATTTGGATGGGGTAGGATGGT TAGCGCTTTCTGCGGTAAATTCGGTGCTCAATCATCACGCCGATGATGGTTAATGCTTGGTGATCGCT ATTGATAATCGGGTAATCATTATTGAGGGGGACAAGTTCGAAATGCTGTCTACCGTTATTGTCCGTAT AAGTTGGACGATATTTTTTGAATGTCGCCTGATCTCCGCCGTTTTTGGCAACAACAAATTCACCAGGA ACCGGCTCTATCTCTGGATCAACAATAATCACATCGCCAGCTTTAAAGTCAGGCTCCATCGAGTCGCC TTCAATACGCAAGGCAAAGCTGTGTTCTGATAAATCAAGATCAGTAAGAATGTATTCAAGACTGCCA TCAAACGCCTCTATCGGGCTTTTTTCTGCAAGTGCTCCTGCTTGCACATAGCTTATCAACGGAACTCTC CTGCTGTTCACnCTGCCAACGGCATAAATGCGCCGCCATTCATCAGCCAGTCTGCATCGCATCGCAA TGCTTTAGCTATGCCGATTATATTACGCGGCTTAAGCGTTTTCCCATCCTCAATGCTTTGCCAGGATTG CTGACGAATACCAGCCTTTTCTGCTGCTTCTGTTTGCGTCAGGCCAAGTTCAATCCTTTTCTGTTTTAC GCGATCTGCAAGGCTCATAAATTCCTCTCTTCATATGCCCTGATGGTCACAGTAAAAACTGTATTTGA CAAACAGAAATAGCTGTCAGAGAATACAGATAAAACTGTGGAGGGCATATGGAAACAATTTCTCATC GCCTCAAACAAAAACGTGAAGAGATGAACCTTTCTCAATCCAAGCTGGCCGCTTTGGTTGGCATGAG TCAGCAATCTCTTCAGGCCATCGAGGCTGGTGCTACGAAGCGCCCACGTTmTGGTTGAGTTGGCTC GCGAGCTTAACTGCGAACCGGAATGGCTTCTTTTCGGAGACGCCAAAAATAAATCATCTGCCGCCTG ACCGGCGGCAACAAACCTAACACCAGAGGAAGTATCGCAAATGGACACCTTAACAACACGCAACAA ACTGGAGGCCCGGAGGATAGAGAGCTGGTTACACAGCCAGATAGCAGAACTGGGTACGACAAAAA TCGCTGAAGTGGCTGGCGTTAACAAATCGACGGTGAGTCGCTGGCGGGAAAACCTGCTTCCGAACA TGTCGCTTTTGCTGGCCATCCTGATTTCAAACCGGGATGAGACGAAGGGTGATTTCGAGGCATGACA CCATGAACCACATCGAATTTATCGAAAAGAACGTGCGAGAGGAACTTATCCGCCAGGGCTTCGCGGT ATCGGTGGCTCAGGGGGGGGCATGGCAGGCAGTAGATATGTACAAGCGAATGTCACAGGCCAGCA Attorney Docket No. PIV-00053 WO

[0293] AGAAAGGGGGAATGTTCGATGACGTTATGCGCCATGCAAAGCTCTGGGCTGAGAAACAAACCAGTT CAACTGAGCGAAGCGAGAAGAAGCGCAGCAAGCGCGGAAATCAGCAGGGACTGTTTTAAAAAGGC GAAAGCCGCGGTGCGCGAACACCAACGGCTCTCAGTGCGAAATGAACGGATCAATTCACAGGAATA ATTATGAGTTCACTCTACCAGCTTTACAAGCAGAAAGACAAAAACGGCACCGGAACCGCCGTCAACA AGACTTATATCGTGCCGCTGAGCGAGTTGTATGTCGAGCCAGGCTTTAACGTGCGTGAGATTGACCA GGCGCATGTCGATGAATTCCGCGATGCGTTCATTGCCGGTGAATTCGTGCCACCGCTGGCCGTTCAG GTGACTGAGCAGGGCGTAAAAATCATCGACGGTCACCACCGCTATTACGGTGCTCTGGCAGCTACTG CCGCTGGTACCGAAGTTCCACGCATTGAGTGCAAAGATTTCGTCGGATCAGAAGCTGACCGCATCGC ATTCATGATTACCAGCAGCCAGGGCAAAGCGTTATCTCCGCTGGAACGCGCCGCAGCTTACCAGCGC TTGGTTAATCAGGGTCGCACTCCGGCGGAAATCGCGAAGATGGTTAAGCGTTCTGTAGCCGACGTTG ATCACCACCTGCAACTGCTGTCCTGCGGCGATGAGCTGATCGACATGGTGAAGGCTGGTGAAGTTTC AGCGTCCACTGCTGTCGCTCTCTCACGCGAGCATGGTGCTCAGGCTGCTTCCGTGGCAACCCGCCAG ATGGACAAAGCCAAAGCCGCTGGCAAAACCAAACTTTCCCGCAGCGCTGCCATCCCACAGTTATCCC CGGCGCGCGCCCGTCGTCTTGCTGAGCTTCTGGCTGATGCAGAAATCGAAGATAACCGCCTCACCGT GGCCTCTACGGCCATGGAAGAGGTTATGGCAATCATCGGTGAGCAAAAATCTTTGATGCGCGAGAG CGGCTGGGAGGAAGCGTGAACACACCTAACGAACACAAAAATATTACCCGGGCCAGATTCGCCAAG ACGGTGAGTCCTGTAGCGATCGCCAGGCTGCGCGCCATCCTGGAAGAACTCAAACGCAAGGAGGCT GGCCGTGGGTAACGTATCTAACTTAGCCGAAGCCAGAGAGGCCAGAAGGCTCCATCAACCGCGTAC AGAAGGCGGTAAGGGGTTTGCCTTGCTGCACCGTAAAATCATGGATGTCCCGTTCTACAAGGACGC GGAGGCGTCACATTTGTGGGTGCATCTCATCCTTAAAGCCAAGCACGCTCCTGAGTCAGTTCTCACTG ATATCGGCGAAATGCTGGTTAAACGGGGACAGTTGCTCAGTGGTCGAAACGCCCTGGCATTTGAAA CAGGCCTGAAAGCAGATCGCGTTCAGTACCTGCTCAGAAAGTTCCAGAAGCTGGGGATGGTGAGCT GGGTTTCACACGGTAAATTCTCTGTTTTTACCATCGTGAAATATGACGATTATCAGTCAAATTCTGTAC CAGCAGATTACCAGCAGATTACCACCGCAAAGCCAGATGTGGCGCAGCCTGTAGAGAAGAGTGTAC CAGCAGATTACCAGCAAATTACCACAGATAAAGAAGTTATTAATATCTCTCTTACTAACGTAAGAGAG AGTGCATCAGCGACAGAAAATCAGGACAAGAAAAAATCGTCTCTCAGCTGTGAGCAGGTGGTTGAG GTTTATCACCGTGTACTGCCTGAAGCCCAGGGCATACGAGTACTGACTGACAAGCGTCGCAATCTGA TTCGCAGCTTCTGGAAAAAAGCCGGAGCAGCGAACCGCCAGCTCGGCGGCGCAGGGTTCACCCTGG CAGACTGGGAAGCGTACCTGAACTACATCGCCACCAACTGCCGATGGATGCTGGAGAACCGCCCGG ACAACCGCACCGGCCGCACATGGCGCCGCAAGTCGCTTGAATACTTCCTGAACGTTGACGTTTACGT GAAGACGCGCGAGGGGGCCTGTGATGATCTCTGAAATCATGACCGTACCTCACAACCTCGAAGCAG AGCAGAGCGTTATCGGCGGCCTCCTGCTGGACGACGACAGCAGCGAGCGAGTGCAGAAGGTGCTTT CCATCCTGAAGCCCGAATCGTTCTACCTGCGTGCGCATCAGGTGCTGTTTGCTGAGATGCGCGAGAT GTTCCGCGACAACAAACCGGTAGACGGGCTGACGCTGTTTGATGTTCTGGAAAGTAAGGGGCTTAC GGAACAAATCGGCGGATTTGCTTACCTGGCTGAAATCGCTAAGAACACGCCAAGCGCGGCAAACAT CGTTGCCTATGCCATGTCAGTGCGTGAAGCGGCTATGGAACGCTACGGCATCCAGCGCATGACCGAA GCAACCGAATTGCTTTACGCCCGCAATGGCATGAGCGCTACGCAGAAGTACGAGGCAATTCAGGCG ATTTTCACCCAGCTTGCAGATCACTCCAAAACCGGAAGCCGCCGTGGACTCCGCGCATTCGGTGACG TTATGGATGACTGGGTTTCCGATCTGGAGAAACGTTTTGACCCGTCAGGTGAGCAGCGCGGGCTGA GTTCAGGCATCCCGTCACTGGATCGCCTTCTGGCACCAAAAGGCCTGGTCAAAGGTTCCCTGTTCGTC ATCGGCGCACGGCCAAAAATGGGTAAAACCACGCTTTACGGACAGATGGCTCTGAACTGCGCTATCC GCGAGAAAAAGCCTGCGCTGCTGTTCAGCCTGGAAATGCCCGGTGATCAGATCGTTGAAAAACTGG TTGGTCAGAAATCTGGTATCAACCCCAGCATnTCTACATGCCCGCCACCGATGACGCTGATGACACC TACCAGGGAGACTATGACAGCGACTTTAAACGAGCTATTGAGACAGCAAATCGCCTTCGTGAAATCG ACATGCTCTACATCGACGACACACCTGGCATGTCGCTGGCGCAGATCGTCACTGAATGCCGCCGTAT CAAGCGCGAGCGCGGAGTGGTCGGAATGGTGCTGGTCGACTACCTGACGCTCATGACCGCTGAGAA GGCCGACCGTAATGACCTTGCTTTCGGGATGATCACCAAAGGGCTAAAGAACCTGGCGAAAGAGCT TGGCTGCGTGGTTGTGCTGCTTACCCAGCTCAACAGGAAGCTGGAGGACAGAAGCAATAAGCGTCC GCTTCCCAGCGATTCCCGCGACACAGGGCAGATTGAGCAGGATTGCGACTACTGGGTTGGTGTTCAC CGCGAAGGTGCTTTTGACGACTCGGTTCCTGCCGGAGAAACAGAGCTGATCCTGCGTCTAAACCGAC Attorney Docket No. PIV-00053 WO

[0294] ATGGCAAAACCGGAACCGTCTACTGCAACCAAATCAACGGCGCGATTTATGACTGCGATCAGGATGC GGCCCGCATGACCGCGCGCAGCCGTGAAGAAAAGCCGGCAAACAAAAAAGGGGGCTTCTGATGAC AGGCAGAGAGGCAATTGAGTGGTACCTGAAAGAATACGGCTATTTCACATGCGAGCTGGTTGCAGA GGCTACCGGAGTTAAGAAATCAGCAATCCAGGGAGCGTCTCACAAAATGAGGCTGGCTGGTGAAAT CGCACTGGATCGCCGCGTATGGCGCACCTGTTTCTATGTCCCTGTTGAGGTTGACGACGACGAAGGT AAGCCGGTCGATCGTTCAGGGGTAAACACCATTTTTGATGAATGCCGCCATAACTGGCAGGGCTATC GGATTCATAAAATTTGGGGCTCGGGAGCGAGAGTATGAAAAAGCATACCAACAATAAAATATCTCAT GAAGAGCTTTTGAATGTCCTGCATTACGAACCTGAGAAAGGGGTATTTACCTGGAAAGTCGTAAAGG GAACAAATCAATCAAGGGTTGGGACTGTGGCGGGTAACTTACTTAAAACAGGTTATATCGTCATTTG TTACGACTATCAGGACTACTTAGCCCATCGGTTGGCATGGTTCTACATGACCGGCTCTTGGCCCGTCA ACCATATAGATCACATCAATCAGGTCAGAAATGACAATCGCTTCGCAAATCTGCGTGAAGCAACACA GTCGCAAAACCTTGCGAATGTCGCTAAAAACCCACGAAACACCTCAGGTTTCAAAGGGGTCGCGCTG GATTTTAACGGTAAATGGCGCGGCTATGTGACTAAAAACTATAAACAGCACTGTAAGTCTGGCTTCC AGACTCCAGAAGAAGCCTCTGCATGGGTGAAGTCAAAAAGATTGGCATTGAATGGTGAATTTGCATC TGATGAGGGGAGTGCACGGTCATGAGCAAACCAACATACGAAGAGTTAGAGGCGCAACTGCATGC AGTGATGGCGGAGCTTAGCGCAGTTGATGCAGTTCACAACGAGGCAGTATTTATCACTGATGAAGAT TACGACAAATGCCCTAAACGTGTGCAGGACATTATCCGCCACCTTGCGGTAATGCAGATTCCAGCAT ATCAGGATTTTGTCGCAACCCTGCGTGCTGAAGCAATGGCTGATGGTATTGAGCAGTGGGCTGATG GTATGGCTGATATTAGCGAATATGACCACATCTACGCATGTGACGCGCGCAATTATGCAGAGAATCT CCGCCAGTCCGTGCAGGTGAAGGGGGTGCAATCGTGAGTACAGGAATCGAACTGATGCAACACGCC CTCGGCATCAACGAGCGCCAGCGCCAACCATATCGCAATTACTTCCTCGCAGATGGTGATCACCCTG ATAGCACAGAGTGGAAAAAACTGGTTGAGCAGGGATTTGCAAAGTCGTCCCCTGCGCCAGCGTGGT CATGCGGTGATGTTGTTTATCAGGTGACAGATGCAGGCCAGGCGGTAGCCATTTCCGCGCTACCTGA GCCGAAGAAGCCAACCCGCTATGACGAATACCTGCATTCCGAAGTTTGCGAATCGTTTGGCGAATGG CTCGGCATCCAGTTGCCGGAATATGAGTGCCGCACAGTTGACCGATTCAAGGGGAAATACGAGTAC CGCATGTTCCGGCGCTCTCGCATCTACTGGGATAACTACGTCGAGGTGAAGGGTGACTGGTGCACCA CTAAAAAAGCAGCCAAGGCCAGCTACAAAGCTGCGCTGCGGGCGCTGAAAGGCAACGCAGGAGAA GGCCATGAGTAGACATGATTTCAGGCGGCCGATAGCGGGGCATATTTTAAGCATTCGTGAAGCTCTG GCAAAAGTACCAGCAGATAAGCGTCTTCACGTTATTGCTGAGGCGCTGTACGACATGAATCCGACAG GAGACGATGAGGTTCTTCAGGTTTCCTGCGGTTGCTACGAGTGGGATATCAGCATGGACTACCGTAG CGCTGACGTCCGTTACGCAGATAAAACCAAAACAGGCGGTGCAGCATGACTAACAACGACGAGCTA TGGATGAAGCTGGAAGCTGCCGAAAGTGACGAACTGGCGCTGAATGGGAATGAAGTGTCTAACCA GTTACGCCATCTGGCTGATAACGAAATCGACAGCGATTCGTTTGCTGTGGTTAGCGAAAATTATAAC GGCAGCGAAGTTGAGTTCGAACGACCGATAACTGATCTGGCAATTGATGCCGCTGAAGTTATTGATG CCCTGCTGGCAGAGCGTGACGCCGACAAGAAGCAACTCGACCGACTTGTCGCAGATCGCATGAATTT GCTTAAAACGATGAGCGTTTATGTCCATGCCTATGAAGAGGCCAAATCACACATAGCCGAACTGGAA GCGCGCACAGTGAGCGTGAAGTTGCCGCGCTATGACCTAGACATGAGTGACTGTGATTCATGCGGG CAGGATTGCTGCGCTGACATGTCCGAAGAACCAGAGGGTGATTATGTGCTGCTGACTGACGTCATCG AAAAGCTGGCCGACGCTGACATCAAACTGCAAGTGGGGGAGTAGGTATGTCGAAACTTACTAAATA TCAATTTGAAACACTGAAAGCTATCGATAACGGTCGCGTAATGCTTCGAGGTGCTTATGACCGCTTCT GGTGGCAGGATGATGACAATCTTTGTAGCGCCGTGGCTCACCGTTTACGATCGAAAGGTCTGATTAA AACCGTCTACTTAAATTCTGTTCGTGACGTCGTGAAAATTACTGACGCTGGCCGCTCAGCATTGAATC AGGAGGCCGTATGACCACAACACAAACGATCAGCCGGGAGCGTCTGGAAGACCTTAAACATCGAAT TTCAATGTGGGGACACGGAGCAGGCTACACGCCGGAAGAAGTTTATGCGCTCATTGAAAACGCTCT GACGGCAATGGACAGCGAGCCGGTGGCGTACTCTCTATTTTTCAGAAACCTGGATGGTGTCCCGGAA ACCAAGGTTAACACGGCAACCACTTTCAACACGAGGATTGAAGCCGAGAATTATGCGCCTGGCGGC AGATACGTAACTCAGGAAGACGGGCGAATCACATTCGTGCGTGATCCGGCTCrTAACCCTGTTGTCG TTCCGCTCTACGCCGCACCGCCATCTCCGGTATTACCTGATGGTCTGCGACTGGCATTGAGCAATGCT GGTATTGCAGCGCCAGAATCAGATGAAATGCTGGCTGCAACAAGTGAAAAATACGTTCAGTTGCTTG TCGATTGGGTAAAAGACAGGAGGCCGTTTACTCCAGCGCCGGTGGCTTATCCTGAACGGCTACCGTG Attorney Docket No. PIV-00053 WO

[0295] CCCGGTACTCCTTGAGCCTGGCATGCGCATCGGTAAGGGCGTGAAAACCTCAGTCCTGCTAAAGGCT TTGCAACGCCGCGCCGCATTCCATGCGGAGCTGGATGCTATGACGCCGGAACAGCGCGCTGAGCAT TACGCCGGGGTAAATGAAATGCGTGAGATGCTTGGCGTTCCGCCGATGCCAGCGCCAGCACCAGCG CCGGTGGCTGTGCCTGATGTTGTTGGTAAATTGCTCAATATTCTTGACCGGCACACCGTGGAGTTAG GATGTGGTGAAAAACCAATCACTCTGGCAGATTTAGTAATCTCACGGGATGGTCGTGAAGAACTGG ACGCCTGCCGCGCCGCCATGCAAGCCGAACCTGTAACGGCGGCTACGGTGCCGGATGGGTGGAAGC TGGTTCCGGTTGAGCCGACAGAAAGGATGGTGATTGATGGCTTTGAGTCTGAGCCTGATGAAACATT CAGTGAGCCGGAAGTATGGGAAGCATATCAGTCCATGAGCGGTTGTCGACAGGCAGCGCATCGTGC CCGCCTGTGTTATGCGGCAATGCTAGCAGCAGCCCCGGCAGCGCCTGAGCAGGAGGTGTGAGATGG ACGCATTTAAAGACATGAGCATTACAGATTGGCTGTTTTTCTCAACCATGGTACTCGTGTGGCTTCAC ATGGCCTACAAGGCATATCACTGGTTCGGCACTTTCTTGCTTCGCCGTGGCTGGCGGTGGTGGAACA GGAAAGACGATAAAGCGCTGGCGATGGATTCTTTCTATGAAGCGTTCAATCTTGCCAGCCTTGAGCC AGGACATACGCTGACTGCAAAAACCGAAAGTGGACTAGTAATACAGGTTTATCGGACAAAGGCGGT GCGCGATGCCTAACCCATTCGACGCCTTGATGTTCGTCCTGTTGGTGGCTGGCGCTCTCAATGAGCTT GTGTGGCTGCCATGGTGAGTTAAACGTCAAGCAACATTGATAATCAATAATCAGTAAGCCATAATTA AGGTGCATTCGGATTGAGCCCCGATTGCACCTTTGCGCATTTAAGGGGACTTAAATGCGACCACAAT CTGAATTCTTCACCTTGTCACAGATGCTTCGCGGCACCTGCGATTTTCTTCATTCTGCGGTTTCCGTTA AGGAGGCCGTATGAACCTGCCAGCAGACGGCATCAAACTCCATCGTGGCAACTTCGCCGCCATAGG CCAGCAGATTCAGCCGTTGCTGGATGCAGGCCAGTGCTTTCGCCTTCAGGTTAAGCCATGGCGCGAG AAGCGTAGCCTCTCTCAGAACGCGCTCAGCCATATGTGGTACACGGAAATAAGCGAATACCTCATTG CGCGCGGCAAGACCTTCGCAACCCCTGAGTGGGTTAAAGACGCGATGAAGCACACCTATCTTGGCTA CGAAAGCAAAGACCGCGTTGATGTTGTTACTGGCGAAGTTACCACCGTTCAGACCCTACGACACACC TCCGATCTGGAAACGGGCGAGATGTACATCTTCCTCTGCAAAGTCGAAGGCTGGGCGATGAACATC GGTTGCCACCTGACAATCCCGCAAAGCTGTGAATACCAGCTTCTGCGCGAAAAGCAGGAGGCATGA TGCATAGCCCACTCGCAAAAGTCATAGAACGCGGCATATTCCGCATTCCTGCGCGCCGGAAACGTAA GCCTGAGCTTAAACCCTCTGAAATCCCAACCCTCAAGGGCTACACCGCCCGTCTTGTCGATCATAAAT GGCTGCGCCTTGCAGCGAGGAAAAATCATGTCTGATTTACGGAAGGCCGCGCGTGGGCGCGAATGT CAGGTGCGAATCCCCGGCGTCTGTAATGGCAATCCTGAAACTTCTGTGCTGGCCCATATCCGTCTGG CTGGCTTATGCGGCACAGGAATCAAGCCCCCTGATCTGATCGCCACCATCGCATGCAGCAGTTGTCA CGACGAAATCGACCGCCGCACCCATTTAGTTGATGCGGAGTATGCAAAAGAGTGCGCGCTGGAAGG AATGGCTCGCACGCAGGTTATCTGGCTTAAAGAGGGACTCGTAAAATCATGAACGAATATCGGATCA GTCTCCCGTGGCCGCCAAGCAACAACCGCTACTACCGGCATAACAGGGGGCGCACCCATATCAGTGC TGAAGGTAATGCATATCGTAATGCTGTAGCACAGATCATCACGGCGGCGTTACTGGACATCTGCACG AAATCACCTCTGCGTGTGCGCATCGAATGTCACATGCCAGACCGGCGTCGCCGCGACCTGGACAATC TTCAAAAGGCCGCTTTCGACGCGCTGACCAAGGCCGGATTTTGGGCTGATGACGTTCAGGTTGTCGA TTATCGGGTAGTGAAGATGCCTGTCGTCAAAGGTGGCCGGTTGCAACTGACCATCACTGAGCTGGA GGTCGCATGACTACCGAAACAGAAAAAGAACTCAGCAACATTGTTGCGTTCCCAACTAAAGAAGATG AGCCGCGCAATCTGACCAATTTTTTGTATGAACAGTCTGAACGGCCTTTCTGTCACCACCCCATGGTT CGGGTTAATGAAAAAGACCGTGAGGTTCGTTGCAGGACATGCGGTGCAGCCGTCGATGCTTTCGAC TGGATGCTTTCGGTAGCCAAAAAAGAGACGCAACTCGCTGACAACGTGAAGGCATTAAGGCGGGAG GAAAAGGAGCGGCGCGCGAATATCGAAAAGCTAATTCAAATTGAGCGCAACGCAAAAGCCAGAATT CGGAGGGTGCGACGTGACCAGGGCACAGATTGATCGTTACCAGGCCGAAAGCGTAGCACGGGCCA AATTGCCCGTCATATGGCGCCACAGCAAGCCAGTTAAAACCACACAGCAGAAACAGACCACAGGGG CAGCCGCATGAACGAACAATATCTGCAATACGTCCGTGAAGAAATCGCGCTGGCTACGGCTGATTTT AGCGGTCGCACTAAGGGGCAATTAGTCGCTCTGGTTGAGCAACTCCAGTACACGAATGAACGCTACC CGCGCAAACGTCAATTCGTCGTTGATGAAGTGACAGGTAAGAAAATATTGCTGCGCAATCCGCCGGT ACCGGGCAAGCAGTCACACGCGAAAGGTACCTCTATCCCGCAGGTAATGCCCGTGGAGTTTTCAACG GCAAGCTGGCGGCGCGCTATTGGCAAACTGGAAGACGCGGAAAGCGCTTGGGTGAAATGGAACTA CCTGCACGATACTGATTTCAGTTTACAGACCGCCATTGTCCAGCATGGGTGGCTCCAGTTCTGCGAAA CAATTAAAGGGCAGCGCATAGCAGGGAAGACCAAAGAAAAGCTCAAGGCGCTTATCTGGCTGGCCG Attorney Docket No. PIV-00053 WO

[0296] CTCAGGATGTAAAAAAGCAGGTCGCCAAGCGCGATGTGTACCTGCAAACCGAACTGGCGCAATTCA TGGGCGTCACTGATAAGAACTGGGGGAACAACTATCGCGATTACTGGCGGGCTATGCGCGGCGTGT TCTTCTCTCTGGATCGTGAGTCGCTAATTTCGATAGTCAGATCACGTTCGCAACAAAAAGCAGCTTTT TCGCATCCAACTATTGCAAAAGTAGATAAAGTGAGTCATATTTGAGTCTACTTTGATATGCTGCCTAA ATTGTAAACGGCGGCAAAGACTAAAAGCCTCGGCATCCCGCCGGGGCTTTGTCATTTTTGGGTCAGA AGCACAGCGGTTGTGCGTTCGGCTGTTAACCGAATGGTCGAAGGTTCGAATCCTTCCTGTCCCGCCA AATTAGCCGGTCTAGTTCAGTGGCAGAACGGCAGCCTTGTAAGCTGCGCGCCAGAGGTTCGATTCCT TTGCCCGGCACCAGAACCCAACCAGCGGGTACCTTCGGCCACGATGCCGACATTGCCACACCCTCTA TTCCCGCCATGTGCGGGTTTTTTATTTCAGGTTCCGGCCAATCAATTCCAGATGACTTTGTCGTTAATG CAGGCCGAGAACCTGACCCTTTTTTAACCACCGAGCGGTGGAGACTACGCAATGGCAAACACGGAA ATCATTCCGGCTTTGCTTCTCGCGCTCATCATGACCGTCCTCGGATCATTTGCAAAGTTCCAGACAAG GCCAGTAAGTGAAAAGGTGAGCCTGTCGAGTCATATGGTCGTGTCGATCTTTGCCGGAATGATGAT GGCTCTCTATGGGCTGGATAAACAATGGTCGCTTTATATCCTCGGAATGGGTTGCGGCGCTGCCGGT TGGCAGGGGGCTGCAATTCTCAAAAAGTTGCCATGGTTCAGCCAAAGCGACAATGGTGATGATTAT GGCAAGAACTGAAATCAGGATCGGGCGATACAGCATCGATCTAATCCCGGTTGTTTTTCTGGTTCTG ACAGCGGTTTTATGCAACCAGCTAAACAGCTCAGAGAACAAAAGCGGCGACGCAATCAAATTGCTCA GCGACGTTAATAGGCAGCGAACAGCGGCAGAAATGCGGGCCTTAAATGCCGAGCGTGAACTGGCA AAAAGCTCAATGCAAAACGCGCCAAACGTTGTAGTGATAACTCCGGACGGAAAGCCAGTGCATAGC GCAGACCGCAAAGAGGAATACAGCGCGGTTCAGCGAGAAGTTTCTTTCTGAGGTGAATCATGACGA AAGACCAGTTCATGCGAGCGGCTGGAATATCATCGGCGCTGGCTGATAAATGGTATCCGCACATCAT CAAAACAATGGATGAGTTCGGCATAAACACGCCGCTGCGGGAGGCCCATTTCCTCGCTCAGATTGGA ACAGAATCGAACGGCTGGCGATCAGTGGTCGAATCGCTGAATTACTCAGTGGCTGGCCTGGCAATCT TCGGTGATCGCCTCACAGCCGCCCAGCGCGAAAAGTTAGGGCGTAAAGTGGGTGAACTTGCTCTTTC TCCTGCCCGGCAGGCAGATATTGCCAATATCGTTTACGGCGGTCGCTATGGCAACACTCAACCTGGT GATGGATGGAAGTATCGCGGGCGGGGGCTGAAGCAAATCACGTTTCGCGCCAACTATGATGCCTGC GGGAAAGCTCTTGGCATCGACCTCGTTAAAGACCCTGATTTACTCCTTCTTGACCAGTACGCCGCGCG CTCTGCCGGTTGGTTCTGGAAGGATAACAACTGCAATTCATTTGCAGACGAAGATGACATTGTCGGG CTGACAAAACGTATCAACGGCGGGTCCAACGGTCTGGATGATCGCAAATCCCGCTACATCGTCGCTA AACGCGTCATGGTGGCATAAATGAAATTCATCGATAACCTCCCGCAATTGTGGAAAATGTGGTCTGT TCGCATTCTTGCCGTGCTGGCACTGGTTGCTTCTGTGTGGGAGCAAATCCCGGATGACGTTAAGGCG ATGATTCCGGCTCAGTATCTCGGATACATCGTTGCTGGCGTTTCGGTGTGTGGCATTATCGCCCGCGC AATTAAGCAGTTCGACAACAATGATCAGCCAAAATGAACATTCCAGCCGTGTTTCAGCCTGGCTGGA AAGGGTGCGGAATTGCGTTGCTGTTGGGCGCTTTGCTTGGCGCGTGGGCTACATGGTGGGTGACTG ACAATAAGTTCACCGCCGACATCGCCACGCTGAAACAGCAGCACGCTGAAGACCTCAAGGCGATCA GCGACGATGCCAGCGCTCAGCGTGACAAAAACGCGGCGCTTGAGCATGCATTCTCTAATGCCGTAGC CAAAAGCGATCAGAAGCACACGCAGGAGTTAAACAATGCGCTCACTGAAAACAAAGCTCTTCGCGA TGCTGTTACCTCTGGCAATCGTAAGTTGCTCCTCGCAAAAGCCGATCTTGCAACCTGTAACGTGTCAA AAAGTGCAACTTCAGGCACCGCCGGCGTGGGCGATGGAACCCAAATCGAATTCAGTCCAGATTTTG GACGAAACATTTACGATATCAGATCCGGCTCAATCAGTGATCAAGCAAAACTAGACCAGCTACAGGA TTACATCCGAGCGGGTCAGGATGCTGGAGTAATCGCCAGATAAAGAATAACCCCGACAAGGTAAAG ATAGTTCTCTCGTAACGGAGGTGATCGCCTGAACCTGGGCCTATCTTGGCAGCCGGAAAGACGGAA GTTGCTTAGCACCGCTGAATAGCAGAGCGAAAGCTGCGACAAGAATTAACGAAGCTCAGTCAGAAA TGGCCGGGCTmTTATTGCTCCAAACCAGCGCACTCGCGTGCGTATCTCAACAAGAGCTTTCCGTAG TGTGAGTCTGAGACAGGGCGGTGGATTTCATCGTTCCGCTCTTGGCTGCCCATGTCTACGCGAGCAG GCTCATGCCACAGAAAGGTAAATACGATGAATAATCCGTCAATTATTCCGGCATTTGATTrTCGCGAA ATGGTGACGCTCTCAAGTGGTGAGTTAGTTACCACCTCGGTAATGGTCGCCAATTATTTCGGCAAGA AGCATAAAAACGTrCTCCAGGCCATACGCAACCTTCAATGCTCAGCAGATTTCAAGCGGCTTAATTTT AAGCCCACTGATTTCATTGATAAAAACGGCGATGTTCAGCCGATGGTGAATATCACTAGGGACGGGT GCATGTTCCTGATTATGGGATTCACCGGACATGCAGCCTCTGCGATTAAAGAGTCCTATATCGCCGCA TTTAACTGGATGGCAGAACAGTTATCGCGCCGCATGGCAATCGGTGAAGAGCTACAGCATCGATTTG Attorney Docket No. PIV-00053 WO

[0297] CCATCAAAGAAACGCGGTCGAAACTGAAAGGCACGATCGGAAGTCGCCTGATGAATGAGCGCAAAA AGGAAAAGCATGTTCTTGCGGTTGAAGAGCAGCGAATTCACGAACTGGCTAATCCTGATATGGTGG CCATCATGCAGGGTAAAGCAGCATGACCAAATACCTCATAGCCGCTCTCGTAGCGGCTTTTTCATTTT CAGCAGCAGCCAAATCAACATGCCCCGGTAAATTCCTGAATGACAATCCTCCACAGGTGCAGTACGA CCAGGAGATTTGCTTCACTCTGTTCTCTGTGCTCTACAGCGACAAGCTGAAGGATCCGGTTATCAGCG CGGAGCATTTGACGGCGGCAGAAGTTACGGCGGCCGGACAGATGAAGCGCAAAGACAGCTTTCAC GCCGAGCAGCTTGTGCCAAAACAGTTTGAGAGCACGCCAAAGGACTATGCGAAGTCTGGCTATGAC CAGGGCCACATGACACCGAACGGCGACGCACCAGACGTTAAATCGCAGTTTGAAACGnCAGCCTGT CAAACATGACACCGCAGTTACCAGACCTCAACCGAAAGGCCTGGCGCCTGATGGAAGAAGCCGTGC GCAAACAGGTGCTGGCTGAAGGCGATGCATGGATAGAAACAGGCATGATTCCAGGCACAAAGACA ATCGGCAATGGCGTAAACGTGCCGAGCGTTATCTGGAAAGCGGTCAAAGCCAAATCAGGTGAGGCT GTGTATGTCGGCGACAACACTACGGGCACCGTAACCAAGCTCACCGTGGCAGATTTCAAAGCTAAGT ACGGGATAAACCCGTTTAACGAGTAAGACATGGCTACAACAAAGCTTCACGTAAAAATCATCCCTCC ATCGGTAGAGCAGATGCAAAAAATTGCCGATGACTATCTGTCTCGGTACAAGGATGTGGATTTGAGC CAAAGTGAAATTGATGATGTGAAAGATAAAGTTGTTAACGACATCATGACTTTATTCACCTTTGAAGC GACAGCCGCAAATTAATTTTCATTACAGAAGCCACTAATCCAGCGGCTTCGATAATGGCTACTTGCTC GATCATACAGAACATACCCCGGATAGACCGGCCAGTTGAAAAGCGGCATCGTCACCGCCTTCCGGG GTATCAATTGACGACAACTTAGACGAGGTTGATATGACAGACAAACAAAACAGATCGGTTCTTGAAG ATCAGCATGTAAAAGCGCTGATGGGCAAAATCAAGGGTGATCTGGAAGAAATATTAAAAGACACCC ATGGCATTGATAAGGCCAAAACGCACGATTCGTTGCTGAAATATACCAGTCAGTTTCTGGAACCATA CAAGCCAAAACCAAGAGTCGGTGACGTGCTTGATATGGACAGGCGGATCAGGACGATGCTGTCAGA AGGTGGGCACGCACAACCAGCCAGTGATTGGTTATGTACTGAAATATCAGTGAAAGAGGTTTCTGG CGAAGCCGCTACGATGAAAGTAAACGTTGAACTGATAATCAAAACAGAGCCGGCAAGCGAAGTATT CAGAAGATTATTTCAGTAAAACTCCGCTATGCAGTGAGCAGGATGTATGTGCCCACAGGCGGACTAA AGAGCCAGCAGAGGAATATGCTGAAATGGCAAAAACAAAATGGCCTAAGCTGCCGCGCTTCTTTGTT CCTCTGTTCCACTCAGCCAACATTTATCTGTGCCGAAACAAAGAAGAGTGGGATGCCGCTTGCACTCA TCTTGGCTTGGATGCTGGCGGTAATGAAATGCTTGCAGGTGCCACACAGTCATATTGCAACACCGAG ACAGGCGAAAACCTTTACCTGATTGGCGTATTCAATGGTGACTTAGCGACGCTGGTTCATGAATGCG CACACGTTGCATTCTATTGCTGCCGCGATGTAGGTGTAACAACTAATCCGTGCGACGCGAACGAAAC CTACTGCTATTTACTCGACAGGATGTTCAGTCAATTCCTGCCATTCATACAGGAAACCAAAAATGAAA ATTAGAGGATTCCGATGAGCGCTGTCATTGAACTTGGCAGGGAAAAGAAATTCCCCATTACCCAAGA GTTATTCGATCGCCTGAAAGATGTCGTACATGAATATGATGGTGATATCGGATTATGCGAAGCGCTC GGCACCCTGGAGTTGCTTAAGGCAGTnTAATTAGCGAGAGCACTGAAAATGTCTCTCAGGAAAGAA TAAGGTAACAACATGGCAAAACCGGACTGGGAGGCCATTGAGTCGGCTTACCGGGCTGGCTCATTG TCTATCCGCGCAATAGCTGATAAGCATGGTGTAAGTGATACCGCTATCCGTAAGCGAGCAACACAGA ACGGATGGCAGCGAGACCTCACGGATCAGGTTCAGAAAGCAACACGGCAGAAGCTTGTTCGCAAAG AGGTTCGCAACGATGGTTCGCGCGAACAGGTGCGAACTGATGAAGAGATTGTTAACGAAGCGGCTG ATGAGGCGGCAGCGGTAGTCCTTGAGCATCGTGCCGATTTGGCGCGCTGGCGTCGTATCGCTGGCA AGCTTGGCGACTTCCTGGATGACGTGGAGTTTACTGAGGAAAACCATGCCTCTCTGGCTCGTTCGCT CGTTGCAGGTGTAGACGCTCAGATTAAAGTCATCAAAGCCGAGCGTGAAGCCTACAACATCGATAAC GGCGACAAGAACACCGATACGGACAGTATCTCTGACCTGATGGATTCATTGTCTCAGGGGGCTTAAT GAAACCTGAGCATATCAAATTGCTTTCAGACAAACTTTGGCGTCTCAACAATCTCTACTGGATCACCG ACAAAGAAGGCAAGACTGTCCGGTTTAAAATGACCCCGGAACAGCTTGAATACTTTGAAGGCATGC ATACCCGCAACATAATCCTGAAAGTGCGGCAACTCGGATTCACGACCGAGGTTTGCATTATCCAGCTT GACGCCGCGCTGTTTGAGTCTGCCAAGTGCGCCCTGATCGCTCATACACTGAATGACGCAAAGCGCC TGTTTCGAGAGAAGGTGAAATACGCCTATGACCGGCTACCTGCCGAAATCAAAGCGGCGAATCCGG CGAGCAATGACGCATCCGGCGAACTGGTATTCAAAAAGGGTGGCTCTATATATGTGAGCACCTCATT CCGTGGCGGCACATTGCGTTACCTGCATGTGTCTGAGTTCGGGAAGATTTGTGCAAAGTACCCTGAG AAAGCGCGCGAAATCGTAACTGGTGCGTTTGAGGCGGTATCAACTGACTGCTTTGCAACGATCGAG AGCACAGCCGAAGGCCGTGCCGGGTATTTTTTCGATTATTGTCAGGCTGCTGAAAAGGCAGCATTGC Attorney Docket No. PIV-00053 WO

[0298] AGGGGAAACCACTATCCCCGCTCGACTGGAAGTTTTTCTTCTTCTCATGGTATGGCAATCCCGACTAC GCAATCGACCCGGTAGAGCCACTACCGCAGCGCCTGACCGATTATTTCAACGGCATTGAGCAAAAGC TCGGCATCACGATAAACGAGCGCCAGCGCGCATGGTATTACGCCAAAGAGAAGACCCTTGGCGACG ATATGAAGCGCGAATATCCGTCGACACCGGAAGAAGCATTCCAGCAATCGGTCGAGGGCGCTTATT ACGCTAAACAGTTCCACTGGCTTTATGCCAACGGCCGCATTGGTCAGATCCCCGATAACTCACACCTC CCGGTTCATACATATTGGGATTTAGGCGTGAGCGACTCAACGGCTATCTGGTTCGTTCGCGAGGTTG GCAACGAGTTTCACGTCATTGACTACTACGAAAACAGCGGCGAAGGCCTGCGGCACTATATGAAAAC CCTCAGAGATAAGGGGTACACATATGCTGAGCACTGGGCTCCCCACGATATTGAAAACCGTGAGTTA GGTAGCGATGCGAAATCACGTAAGGAAATCGCCAGAGAAGGCTACGAAATCGACGGACAGCGCTA CAGCATCAAATTCAACGTTGTTCCGAAAGAGTCGGTAGATACCGGCATAGAGGCCGTGCGCGAAAT CCTGCCGATGTGCGTCTTCGATGAAGCCAGATGTGAAGAAGGCATTAACGCTCTCGAAGGTTACCGC AAGGAGTGGGATGACAAGCGTGGTTGCTGGAAAGACAAACCCCTTCATGACTGGACATCGCACGGA TCTGACGGATTCCGTTACTTCGCCGTGGTCAAAAACAACAAACGGACGTTCAAAACGAAAACGCGTC ACTTTTCTGCATAACTGGAATTAACTATGACCGATTCTGTAAGAGTGCGATCAGCGAAAGTCGAGGC TATTGCAGAATGCTGGCCTATGATTACGGCGCTGCTGGGTGGCACGGAAGAGATGCGTGAGGCAGG AAAATTATACCTCCCGCAATGGCCAAACGAAGATAACAATTTCTACCAACGCCGCCTGGCGGTGTCG ACGCTGCTACCGGCATTCGCCCGCACAGTTGAAGTGCTGAGCGGAAAACCATTCTCCCGCCCGGTTA CATGGGGCGACGATGTGCCAAAAAGCATTGCCGATCTTTTTGGCGACATTGATATGCAGGGCACAAA TCTCCATTCTTTCCTCGCCAGCGTGCTTGAGGAAGCCATGGGATTCGGTATCTGCATTGTGCTGTCTG ATTACCCTCCGACTAGCGGTGAAATGACAGTTGCTGAGACAAAGGCATCTGGCGTTCGTCCATATTG GGTAAAAATCTGCGCTAACAGCTTGCTTGACTGGCGATCAACGCGTATTAACGGGCGTGAGGTGCTT ACTCATCTGCGTTTTGTTGAATGCGTCACTGAGCCGGATCCAGCAGACGAGTTCGGCGAGGTTCACA TTGAGCAGGTTCGTGTGCTTGATGCTGGACAGTGGCGAATTTACCGCAAGAAGAAAGATGAAACGA ACGGCGAAATCAAGTGGGCGCTTCATGATGAAGGCACTACCAGCCTGAATTATGTCCCTGCGGTGCC TGTTTATGGCAAAAAGCTCGGCTTCATGCGGTCAAAGCCACCGCTGTCTGAGATTGCCTATCTCAACG TTGAGCACTGGCAGTCAAAGAGCGATCAACAAACCATTCTTCACGTTGCCCGTGTTCCTGTGCTTTTT GCCAAGGGCTTCAGTGATGATGATGAAATCACTATTGGCTCATCAACCGCGGTTACTACCAACAACG ACAAGGCTGAACTTCAGTACGTCGAACATAGCGGAAAAGCGATTGAGTCCGGCAGCAGAGACCTTG AACGTCTTGAAGACCTGATGCGGCAGATTGGTGCCGAGTTACTCGTCATCAAGCCTGGCCGTGTCAC GGTAGCGCAAACCCGGCAGGAAGACGAGGCGGGGACATGCGCTCTGCAACGCATTGTGCAGGACT TCGAAGACGCAGCCGACCAGATGCTTCAGATCACCGCAGAATGGCTTGGAGAGAAGTCTGGTGGCT CGCTCACCATCTATAACGACTTCGGCGCATCTTCGCTTGCCGAGGCCACAGCGCAACTCCTGCTCGAT GCGAATATCGCTAACGTCATCTCTAACGAGACGCTGTTTAGCGAACTGCAACGGCGCGGGCTGGTGA AAGACAATATCAAATGGAATGAGGAGGTTGTGCGCATTGCTTCGCAGCCTCCACGCTCTGAAAGCAC CAAAACAAACCTCAACGCGTAAGCGTTAACCACGAACACGGGCCCGGCTAACCACCGGGCTTTTTTT ATGGCTGAAATCTGCGGATGCAGAACAGCGAACCGGGCCGGATGGCTCACAGCAAGGTTGGATGA CCAGATATGAAACTGAAAATCGATGAGAACGGCCACGTAGTAGTCAGCGATGGCAAGCCTGTGTAC ATCAACGATGAAGGCAAAGAAATCGCCTTTGACGTTGCCGGAACCGTTGCAACCATCTCCCGCCTTA ACGGTGAGGCAAAGTCGCACCGCGAGCGCGCGGAGGCCGCTGAAAGCTCTCTGAAAGCTTTTGACG GCATTCAGGATCCGAAGGCTGCATTGCAGGCACTCGAAACCATCAAAAACCTCGACGCCAAAAAACT GGTGGATGCCGGTGAAGTCGACAAAGTCCGCTCAGAAGCCATTAAAGCCATCGAAGAGAAATATGC GCCGATCGTTAAGGAACGCGATrCACTTCAGGCGGCTCTGGTGAATGAGAAGGTAGGTGGCAGCTT CCTGCGCTCTCCTTTCATCACCGAGAAAATGGCTATTCCGGCTGACATGGTGCAGGCGCGCTTTGGT GAATCCTTCAAGCTCGAAGGCGATCAGGTTGTGGCTTATGACAAGCAAGGCAACAAGCTTTTCAGCC GCAGCAACCCTGGCGAAGTCGCCAGCTTTGATGAAGCGCTGGAAATCCTCATCGATAGCTACCCATA CAAAGACCATATCCTGAAAGGTTCGAATGCTTCTGGTTCGGGTTCTAAGGGCGGCGGTTTTGGTGGT GGCAAGAGAACAATCACCCGAGCTCAGTTCGATAGCATGTCACAAACCGACCAGGCAGCAGCGGCC ATGTCCGCAAAAAAAGGCGAAGTGCTCATCGTGGATTAACGAATGCCGCAATGAATGATCATGCCA GGTATTACTCCTGGCCCTGAAATTGGAATACCCCGGATGGGGCCCGCGAATGAGCTGGATGGCTCG ACGCAAAGAAAAATTCACTCATTTAAACAAATCTGGAGATGTGCCTCATGGCAAACACTTTAACGGG Attorney Docket No. PIV-00053 WO

[0299] TCTTATCCCTACCATTTACGAAGCACTGGACGTTGTTTCCCGCGAGCAAATTGGCTTTATTCCTGCGGT ATCACGCAATTCCAGCGCTGAACGTGCAGCTTTAAACGAAACAATCATGATCCCTATTGCCCCGCAG GCAGCGCTGGCAGATAACACCCCTGGCGTGACTGCTCCGAATACCGGGGATCAAACTATCGGTAAC GTCACCATGACCATCAGCAAATCAAAGCACTATCCGATTCGCTGGAATGGTGAAGAACAACGCGGCA TGAACAACGCAGGAACCTATAACGGCCTGTTGCTTAATCAGTTTTCACAAGCGTTTCGCACCCTTTCA AACCAAATTGAAGTCGACCTGTTCAATACCGCGTATCTTGGTGCGTCTCGCGCTTATGGAACCGCCG GTACTGCGCCGTTTGGTGTGGCAGGCGATCTTTCTGATATCGCTCAGGTGCGTAAGATTCTTGATGAT AACGGTGCGCCGCAATCTGATTTGAATCTTGTTTrGGGTTCAAGTGCCATTGCTAACCTGCGAGGCA AACAAAACGTTCTGTTCAAAGTTAACGAATCAGGCACCGAAGATTTGCTTCGCCGCGGCGTGATTGG TTCTCTTGAAGGTCAGATGATCCGAAACTCCAACGCCGTGCAATCAGTGGCAAAAGGTACCGGTTCT GGATATACCACCGATACTGCTGGTTATGCAGTTGGTGCTACTTCCATCACTCTGATCGCTGGTACAGG AACCATTCTTGCTGGCGGGACGGTAACCTTCGCCGGCGATTCGAATAAATATGTCGTAGCCGGAGGA ATTACCGCTCCTGGAGTTATCACCCTGGCCGCTCCTGGGCTTATGCAGGCAATCCCGGCATCGGCTAC AGCAGTCACCCTTGGCGCGTCTGCGACGCCTAATATCTCTTTCAGTTCCAGCGCTATCCAACTGATCA CCCGGTCACCGCAAATGCCTATCGGCCCGGACGGGCGCGCCATGGATATGGCAGAGGATGTTATTC AGGTAACTGATCCGGTATCAGGCATCGTGTTTGATATTGCCGTTTACCGCCAGTACATGCAGTTGGTT TACCACGTACGACTCGCATGGGGTTGCCAGGCCATTAAATCAAACCACATCGCCATGTTGCTTGGTTA ATAAAATTAAGGGAGGCAATGACTGTCTCCCTTAATTGAGGAAAAAATATGGTTATCGAAACTATCC GAGTTACGTCTGAAGTTACTGAAGAAAATCCCCTTGGATATGTCGTGATTAATAAATCAGACCTCACA GAAGAACACGTAGTTTTTGAAGACTGTTCTGATGCGAGCGCGGGCAAGAAAAAGGGCAAAACCAAA ACCGTGGACGCTACTTCAGAGGGCTCCTGAAATGGCATTAACAGACGCTCAAATGACTGATGTTCGC CGGTACATGGGGTATCAGCTTAGTGGTACTACTATGCCAATAACTGCAAATCAGGATCTGGTTTACG TAAAATTTGGCATGGTAACAATGTCGCTTTTTCAACGTCTGACATCCCTCAGCCCAAGTGAAGAGAGC GTTCTGATTAATAACTATCTGAACAAAATCAGCATACTTGAAACCGCCATTACAGATGCCAGTGACAA CCTTGATACAGACCAGGCAGCAATCTGGATCCACAATAAAAACGAAGTGAGTGACCGCATGAGGCT TTATAACCTCTGGAGACGTCAGATGTGCGCATTCCTTGGTGCTGAGCCTGGCCCATCTCTTCGTAAGG GCGGTTGCCAGATCGTCAGGGGGTGATGAGTGGACGGCGATAAAATTCAACTGAAGATGTACAGG GGTTACGCCAAGGCCGCGCGAGTGGTTGGCACCTCCTACACGCATTTTCGCCCATCTTCATCATCAAA CCCGATGGATGCAGCCAATCAACTTGCCGATATCAAAATGACGTTAAACGCCTGCGACATGAAGTAC AGCAAGCCAAATGTCTACGGCAAGTCAACGTGGTACGGCATCTTTGACGGCACTCTGGCGCAGCCGT TTGACTACTTCGAAGGCGTAGAAGGCATCTTCTTTGTTGCGGCAATGCAGCAACTGCTTCCGATCCTG ATGGTGAGCTGTAACCGCACGGTGAATGTGCTTCGCCCGTCAATGGAAACCGGCCCCGGCGCTATC GGATACGGCGGCGACACCGAGGGGAATGAAGTGCCGCTAATGACTGGCTGGCCTGCCTCAGT

[0300] Prophage P-5812-02 (SEQ ID NO: 13) TCACATCCAGAATTGACGCTGTCCGGTCAGATTTGGGTGCGGTGGTGCGGGGTCAATTTTTCCCGGA GTAACAATATAGCGCTCTATCGACTCCATCGTCACAAAGGTGCAACTGCAATTAATATTTGTGCACTG GTGATAACGTTCTTTTGTGTTCTCGCTGAGATAGCGGCTAGTACGTGCATGCGCCGCATGCTTACATT TTGGACAATGGAACATAGTTAACCCCTATTCACTTAATGTGAATTAATGATACTTCATCATTCACCATT TGAGAATAAATGTTTACACACTAACCAAATAAGGATGTTAAATGACTCTTTTGAAACTCTCCATTTTTA GCTTATGTCTTTCCCTrTCTTACTCTGCTTTTGCTCAAACAAGCGAAAAATTGACCTTAGAAAAAAACT TAAAAGTATGGCAACCGACTGAAATATCCAATAAAAACAACACAATCACTGTTGTATTACCCGGCAA AAACCTGACGCCGGAAGCGTATGAATCCATAGTTACTGGGGGCATTTGCATGCCTATCTGGACGAAA GACGCACCTAACAAGTACCTGGAAAAGATCAAAGAATTAAATATCGTTAATCAATATAAAGTGCTCG GGTTTACAGTTGAAAGCCCTGCATCTACCTGCGCTGAAATGGGTAAGCTAATGGATAAGCCAGGCAA AGCATATCTTGCATCAAAAACCCGTATGTACCTTCCAGCCAAATAATCGAGACAACCTATAACCCGCA TATTCGCGGGTTATTTTTCAATATTCACCATCATCCGATTCATCGTCAGCACCGGCCTCATACTCAACA TCTGATAGTTTCACCTCAAGCTCTAACGCTGTTGTATAGCCGCGATTGCTCAGGGAATGGGTCACTTT AGTAATGATCCACGCCTGCTCATCAATGATGGTTTTAAAGCCCGAGACTTTCACCGGCGTTTCGGGGT Attorney Docket No. PIV-00053 WO

[0301] ACAGCTCGGCGCGCCCGACCGCAAGATTTATTGAGAACTCGGCAACGCCGCGCTGTAGTTTGTCCCA TTTGGCTGTCGCAGCCCGTACCGCCTGGGCTTTTGTAGCGAACACGGTTGTCAGCGCAAAAACGTTA TCCGCCTCACCGGCCATGTATTCACCTTCCCGCGCCTCGGGGACTTTGGCCTCCTTCTITTTTGCCTTT GCTTTGGGATGCTGCAACGCGCGCAGGTGCTGCTCTTTAGGCTTACGCTTTACCTTCACCTTTTTAGG CTTCGGCTCTTTGGTATGGAGCCATTGCGCCGTCACGCCGGTATAAGCGCCCCGGTCAGCAATCGCG AACTGATGGCGGTCGCCATCGCTGCGGGTGATCGTCACCTGGGGAATCGGCTTGCCACTGGCGTTA ACTGCATTACCCGGTCGCAGGAAAAGTAACTTACCGGCTTTGATGGATACCTCGCCGCCGTTGCGGT CAGCAAGCCGTGTAAGAAATTTGGCGTCTGACTCCTGCGCCTGGTCGATATGCGGCAGCGGGATTTT TGCCAGCTCCGGCGCAACAGCGGCGGTTAATTTGTTCCGTGCCGCAATGGCCTCAACGATTGCCCCC AGCGTTTTATCGTGCCAGGACTCTTCCCGGCGCGAATTCAGCGACCCGCGAAAATCAGCCGAGCGGG CGCGGATGGTCACGGTATCCGGCGCGCCCCGGTGCTCAACCTCATCAACGGTGAAACTGCCCTTATT CACCAGCGCCGAATCTTTCCACCCAAGCCAGAGCGTCAGCACGGCACCGCGCACCGGAAGGGCGAC AAGCCCGTCGGCGTCGTCGAGCTCGATATCAAGCTGATCGGCTTCAAATCCCCGGTTGTCTGTCATG GTGAGGCTTATCAGTCTGGTGCTGATATTGCCGGTAATATCCTTGCTTTCGAGTTTCAGCATAAAGTC GGGAACAAGGGAACTCCCCGCCTTGCTATAGAGTATTTCGGACATCAGAACCCCAGCGTTGATGTTA ATTTTGTTGCCGCGTCTTTCGCGTTACCGACGAGGGTTTCGGCCTGTTTACTCAGATCGCCATACACC GCCGCGAGTGATTCATCTACCCGCGTTAACGTCAGCGTAAAGCTGATTTTTCGCGGTGAACCATCGG AAAAAAACTCGGTGCCGGTATCACTGATGCTGTTGATAACAAACATGCCGTAAATCATCCCGGAGCC GGAAAGTAACGGCCATGCCTTACCATCCTCAGCCATCAGCCGGAGCGCGGTTAAGGACAGATGGCC GCCTGTCAGTTCGGGATAAAGCTCACCGCCAAGTGTGATTTTCTCCTCGCCCACGCCGAGAAACTGA AAAGCGTCACGTTTACCCACGCGGGCATTAGACGGCCAGCGATATTCCGCGTCACGCTGCATGTTCT GAAACGGCAGTGTCTGACGCATAAAAACGAACAACCCCAAAGCCAGCATCATTTTTCTGTTCCCCTTA CCCGTCAGTTCTCATGTCAGCGCGTTCGCGGGCGCGTTTTTCCCTGTCGGCACGCTCCACGGCATCCC GTAACTGATTCCCGAGCTCACCACCGGGTGCGCCACCGTTTTGCAGGGTGATGTTGTAATTGCTCTG ACGGTTATCAACATAAGATTTACCGGCACCGGGCGCGGTCACCGGCTGATAGGACGGATAGCCGCC ATACGAAGAGGTGAGAGGGTAATATGACCCCGGCGGCGAGGCATTGCCTGGCTGTTCCGACCGGG

[0302] AmCGCCTGGCTGGCTTTTTCGCCTTTCTCGTCAAGACCGTCAGACTCTTTGTTAATCACGCCGAGTT TTTCCAGCACCCATTCAATACCGGATCGCAGTTTGTTGAACGCCTTAAGCGGCAACATCAGCGCATCA GCCAGCGCCTGACCAAACGCGATACCGGCATCCTTGCAGGAATTAAGCTGCTCCTGTGAAGATTTGA CCGGCTCAAGCAGGTTTTTAAACCAGTCCCACGCCGCCTTTAATTTTTCGCCCAGCCAGTCAAACATC GGTTTAAGCGGTGAGAACATTTCACCGACCGGCGCAAAGGCCGCCGTGAGTCCCTCAATCACACCGC TGAAAAAGGCGCTGATAGGCTCCCAGTATTTGTGGATAAGCAGGGCACCGGCGACGATGGCGACGC CAATCGCGATAATCGGCCACGTCAGCGCACCGAGCACCGTCATGATCGCGCCGCCCACGACGCTGAA GACCGTCCCGAGCACACCGGCAATCGCAATAATCGCGTTGATGCCCGAAATAACGGGAAACGCAAC CAGGCCGATAGCGCCGATGACGGCAATCACGGCAGTTGCCACCCCGGCCAGCACGCCGAGGGTTTG CGCCAGCCCCTGATTTTTCTGGATCCAGCCGTCAAGTTGCAGTACGTAGCCGGTCGCTGTCTGCACCA GTTTGCGAAGGGACGATTCCTGCTGGTCAAACAGGTCAGTACCGACCGCCTCGTAAGCCGACTGGA ACTCTTTGAAGTCGCCGCCGAGGTTGTCCTGCATGACTTTGACAAGCTCCTCGGTCTTCCCGTCAGAT GCTTTTAACGCCGCCGTCAGTTTATCGAGTCTGCCAGATGACGCGCCCTGTAACAGGGCGTTAGCTG ATTTAAGCGCCTCTTCGCCAAAGATGGTTTTCAGGTATTCGCCCTGCTGCGCATTACCCAGCTTATGCT TCGCGAAACTGGCGTTTATCTCCTTCAGGATGGTAAAGACAGGCCGCATATTGCCTTTGCTGTCTGAC GTTTTAACGCCGAGCTCTTTAATCGCGCGGAACGCrrCACCGGTTGGAGCCTGGAGGCGGGTTACTA CCGCAGCGCTCCCCGTACCCGCCATCGACCCGGTTATATTGTTGTCATGAAGAACGCCGGTCATTGC GGCAGCTTCTTCGAGGCTTACCCCAGCCGCGCGGGCAACCGGGGCAAGATATGTCATGGCATCACTC AACCCCTGAAAATCAGCCGCCGACTTGTTCATCGTCGCCGACAGGACATCGCCAATATGCCCGACTTT ATCGTTTGATAATTGAAAGGCGCTTTTCGTACCCAGCAGTAACTTCGCGTTCTCCTCCATCGTTTGCCG GTTTGCCAGCGCCATATTCAGCGTGACGGGCGTTGCGGCCTGGATGGCGGCTGCATCGCCTCCCGCT TTAGCGATGATGATTTGCGCCCCGGCAGCATCATCCGCCGAGGCGGCTGTGTTGTCGCCGAGCTGGC GCGCCTGTTTGCGCAGTGCCGTCATTTCTGCGGAGTCTTTCGCTACCCCCAGCGTTGCCTGTAGTTCT GAGTTCTTTTGCGCAAAGTCAAAACCCGGCTTTAACAACGCCGCGCCCGCCAGCGTGGCGGTTGTCG Attorney Docket No. PIV-00053 WO

[0303] CCATTCCCACGCCAGCGGCACCCACCGCCGCCGAGTTCCCGGCCAGCTCCTTTCCGGTCTGGTAACGC TGTTTTACCCGGCTCAGTTTTTCCTGTTGCGCGCTGACTTTTCCCAGCGCCGCGCGTTGCCGGTCAAG CTGGGATGTCGTCTCGCTGATGGTGTTTTTCAGGCGGCGCTCATCAGCCGCCAGCGTGCGGGTATTA ATCCCGGCCTGACTCAGCGCCTGTCGCTGGCGTTGCACCGCCTCGCGCAGACTGTTGTGTTTGAGCT GGAGCGCGGTGGCGCTTTTACGGGCGGCCTCCATTGCGGCCGCCTGTGCGCGTGTCGGCTGCTCTGT GTTTTTAAACTGCACCGCCAGCGCGGCGGCTTCCTGTTTTGCCTTTTCCAGTGCCTGACTGGTCACGG CAAGCTGTGCGCTGGTCTTGCGAAAACCCTCTATTCTGGATGCCTGGCTGTTGAGCTCACGCAACGA GGTCTGAGTCGCCCGGATACCTCCGGCCAGCTCTTTGCTGGCCTTGTCGATGGATTTTAACGGGCGC GTCGCCTGGTCAACCGCTTTGAGCAATACCTGTAATTTCACATCACTCATTCGTGTTTCCGCTTCGCTG GAGCGCTTTTTCGCGCCATGTGATGAGCTCGGTCAGGCTCATGGGAAAGAGCTCTGATGGCGGCCA GTGGAAAATCACGGCGATATCCGCCATCAGGTCATCAACCGAGAGATTTTTCGGGAAGGTTACTGTG CCGAACTCGGTGACAAAAAACCGACCACCTGACCGGCCAGCGCAACCAGATCGGGCAGCTCGAGCG CGGCGACTTCCTGCTCGGTCAGGGAGGGGGCTGTCATGCGCGGCAGCACCTTAATCAGCGCATCCA CCTCGGAATTGGCGACCGCCGCCAGGCTGACGCCGCGAAGCGTACCGGCGTTAGGCTTCATCAGCG TAATGGTGTTCACCACCTGATCGCCGCGTTTAATCGGATTTTCCAGGGTCACTACGTTTTCTTTGCTCA TGGTTTTCTCGTTTCAGTTCGTTATCAGGTCACCCGGCAGGCGGTGCTCGCCGGGGAAAAATTACAG GCCAATATTGCGGCGGTGCTGTTCCAGTCGGTCGGTGCCGTTCACTTTCTCAATCATGTTGATGGTGT CGATTTCGACCAGCTCCTTACCGTCCATCGTCAGTTTGAAATAGGTACACACGACCGAAATTTTGGCT TCGGTGTCTTCGCCCATTTTGCTTTCGCCGGTGTCGATTTCTTTCTGGCGGCCACGCATGACCACTTCG ACGGCGACCGTTTCGCCGGTGTCGTCGCGCTGGTAGGAACCGGCAAAACGCACCGGCACCACGTCG ACGCCGGTCGCGCCGTACAGCTCCCAGATAACCGAATCGGGGAAGCCACCGAGCGACCATTCCATC GACAGGGCATCATCATCAAGACCGAGGTCGACCGGCGCACTGCCGCTCATCCCCGCCCCGCGATAGT TCTCCAGCTTGCGGGTCAGCTTCGGTAACGTGATGGACTTCGCCACCCCCTGATAGCTGTAGCCATTC AGGAAGACGTTCATATGTTTCAGTTTGCGCGGCATTGCCATGTGTCAGGCTCCTTAGTTGCTGTTAAC CGCTGTCACCAGATTGGCGAGGTACTTGTCGGTGATGCGCTGGCGTAATGTCAGGTTTTCCAGCGGG GGAACCGGCGTATAGTCGTAGTCGATATAGAGTTTTCCGGCCTTAAGGGTTTCCTTGTCGTTGGCTTC CTCGTCAAACCAGCAGGTCGCATCGACGATATAGCCGnCGTTTTCAGCTCGCGGAATTTGGCATTAA TCCCGTCAACGATGTCGCGGATAAGCGTCGCGGTGATCGGCTTGTCGACCGCCCACATATGCGCCTC CGCCATGGTGTCAGCGATAACCTGTGCGGTGCGGGTGTAGTTCTCAAACAGGAATAACGGATCGTC GGAACAGGTGCGGTTTCCCCAGAAGCGAAAGCCATCTTTGCGAATCAGCGTCGTGACCCCGGCTTG GTTCAGCAGGTCGGCATCGGTGCCGGGCTCCTGCAAATCCCAGAATACCGACGCGCTGATACCGGT GACGCCATTCACGCCGACGTTGGACAGCGTTTTATGCCAGCCGACAGTCTCGTCGATGTAAGCGCGC AGACCCAGCGCACGCGCCGTGGCGTAAGCGGTTCTGGTGGCGTTCGTCGTGGTGTCCCAGGTGAGG AAATCCGGCCAGATAACCATCAGTTCGCGCTGGCTGAAGTTGTCGCGGTATTTGATGGCCTCGGAAA TCGTTTTACAGCCCCACGCGCTGACATACCCGAAAGCGCGCAGTTTCTGACAGACCGGCGCAAGCGC GGTCGCCACTTCGAGCGTGTCGTAACCCGGCACACCGAGAATGCGCGGCTTGACGCCGGTGACTGC CTCGGCAGTCAGCAGGGCTTTCAGCCCGGTATATTTGCCGCTGGCGTCCGTGGTGCCGATGATATTG GAAACCGTCTGTGCGAATGCGGCCTCCTCATCCTCGCCGGTGCCCTCTTCGACACGCACGACAACGG TGACCGGTTTAGCCTGGTCGGCGATGGCCTGAAGCGCGGCGCGCAGGGTGCCTTTnTACCGGCTTT CGCAATGGCGCTCTGTACGCTGGTAATCAGCACCGGCTCATTAAGGGGGAAGGTTGCTGCGTCGGC GTCGCTGGCGGTACAGACCATACCGATAATCGCCGTCGAGACCGTGGAAATAACGCGCGTGCCGTC GTTAATCTCGACGACCTGCACGCCGTGGTGAAAATCACTCATCCGTTTAACTCCGTGGTGTTGGGGT GAGTGATATTTTCCAGGGTGCCGGGGTCAGCGGCTATTTATTGGGGTTGGGGGGAAGATACAACAA CTCTAGTTTAATACATTATTCGTTAAACAGCCATCGACCCGCTTTCGCTGAGGCGATAAGCATGCTCA CTGTCAGCGCTTCCGGCTCATGGTGATCTGTTCTCAGATACCGGGGCAAAATCGCATTAGGCAGAAA TCGAATCCCCTCATTAGGGAAATAGCGGTTGAAGTTAAACGCAGACATATCGACATCCAGTGCAGCG GCATACTCTTCAAGAAAATCGCTGGCATCTTCCGGTAACACGCTTTGTCTGCCCGTGCTTAGCGAGGC ATCTTCCGCCATCGGCCAGAAGTGTTTTTTTATCAGTTCATGAACCTGCGCGGCCTTGTCTGTCATTGC TGCACCCCTGATCCGACAATGAGTTTATAACGATGAATAACATTCTGAGTAATCATAGCCAGATCATA GGCCGTTAAAATCGCCCCCACCCCGGGTAACCATCTGCCTATATACGCACCAATACTTGTCGTATAAG Attorney Docket No. PIV-00053 WO

[0304] CCCACTCCCCCCGCAACATTTTCGCCCAGGTAATAGTCCGGTGTGGTTTCTTAAATCGCTGGCGAATC AACTGGCGCGAAGCGAGAGAAAGCGGGCTTGTGCCTTTCGTATnTTTCCGGCGGCTGCACCTATTT TCCCTCTCACCGGGATAACGGGAAGACTCGCAATCATGGAGGAAACTGCCAGCAAATCGATCACATC GCTGAACTGATTTTTAAACTCATCCAGAATAAGCCAGTAAAGAAGCTCGTCTTTATCCACGCTCATGC CATCAAAGAAGTATTTTCCGTTTAATTGTTCGGTCGTATCCATAGCGTTTCCCTGTTCAGGCTGACGAT ATAAATCCTAATACTGCCCTTTTGTATCGTCCAGAACATACGCAACAGCTTAATCTTAAAAGTTGCAGT GTCATCAGCCCGGATGCAACAGTCTGAATTACCTGCCGCCCGGCAACTCAGGTTGTTAAACATCGTG GGCTGTCGGGCTGTCCGTCACTCTTCAGCGTAATCCGAAATCAAGACCGGTGGCGGAGCGTCAACCC AGACAGGCAGACCGTCCGTGCCTCTTCCGCATACTTTCCCCTCCGGGGGCTGGCCTGTGAATTCAAG AAAAACAGACTCCGCAACTTCAACGCCTGATTCAGGCCAGTTTCCTGATTTAACGTACTTTTCTTTAAG GTTAAAAGGATAAAAAAAGCCCGTTTCGCTAAATAAATAATTCATATTAATAACCCTCCGCCCACCAG TAGAAGCCCGTTGCTACCGGTACTGCAACGGCAGAATTACCGGATATGACAGAGGCGATGTTCGCC ATCCCAAATCCCGTAAGGGTGGGAACAATAAATTGTAAGACTGACTGGACTGTTACCCCCGTGGACG GGCTTACCACGCTATAGCCCATATTCAGTCGTTTAGTCGGGAAGGGGATCGGAAAGGTAACATTCGT TACAGCCCCCGGTGCAGCATTCGCAACGAAACCAAACTGTTTAATCATTCCGTTGGGTAATTCGTAAT AGCCGTTATCACCTGATATATAAAATTTATACGAAGACATATCCGGGATTTGTCCCGCCGCTGTACCG ACATTCTTTTTCGCTGCGCTTCCCAGACCGAGGTATTCGAGGAGAGCAGCAACGCTTTTCCCCGACAG ATCAGTCAGCGTTTTGTCCAGCGGTTGTTTTTTTGTCAGCTCACCGGATACCATCGCCGCAATTGCCG CCTGTACAAAAGCGGTAGTTGCCAGTTGAGCGTCATTTGACGTCTGTTCCGCCGTCGGGGCTTTCGG CGTGCCGGTGAAAACCGGGCTGGCAAGCGGTGCATACTGTTTATGGGGATTGCTGTCGGCAACATG CTGTTTCAGCAGATTATCAGCATACGCTTTCACCTCAATGACCGCGTTATCGACATACTGCCGTGTCG CCAGCACGACCGACGGGTCGATTTTCAGGGTAACGGCAGCGGTCGAGGAGACAATCAGCACCATGC GTATGGTCTGCGTGCGCCCGCTCCCCTCCTGCAACTGCGGTTTGTAGGTTTCCGGGCAGTTCGCCACG GCAATTAACACGCCGTTATCGTCATAAATCCCGATTTCACGGATCCAGAATCCGCCCTCGTTTTCGGG GATGATTTGCTCGGCAATAATCTGACTGGCGTTATTCGGGTCAACGCTCAGCATATTCAGCGGCGCA ATGCGCTTCTGATTAACCAGCTTTGTCTGTGCCGCGTCCGGTGTCGGCAGGGTGCCGTTGCCGTCCCC GACGGCCATTTGCGTGAGATTAAGTTTAGTGCCGAGCGCGGCGGCGTTCGCCAGCAGTGCCGCGCC CTGATTGGTCAGAATTGCAAAAAATTTCGCGGTCATGCGTTAACTCTCAGGTTATCAATAAACAGAAT GGCCGAGGCCGGGAAGTAATCACCGCCAACAGCGATTTGCTCCGGCGTGTAGGGGTAAACCGTCAG CGAATTGCCGTCATAGCTTCCGGCCTCAACATGGACATTGCCGGACGTGCTCAGGCTGATAGCGAGG CCGGTTAAGTGGCGACTGGCCGGTTTGGCATCATCAATAAGCCGCTCCAGCTCCTGATACATTTCATC GGTGATCCCGCCGTCCAGCACACCGACAACAAGCCGGAAGGTTCCAGGCTCCTCGTTGAGCTGCCAC CACTCCTTAATCTCAATCAGAAAGCCGAGCGGCTCGACCACGCGACGTAATGCGCTGATGGTGCCTT TGTGCTGATGGACGTAAAACGAGGCGGCAATAACGTTGCGTTTGGTGGCTTCCGGCCAGGCTGAAT CCCACCGGTCTACTGACAGCGCCCAGGCAAGATAAGGCAGCAGGTTAACCGGGCAGGTTTTCGGAT CCCACAGGGTACGCAGCGGCACCGGCACGCGGATGATTTCCGCTGCGGCGGCTGCGGCGGCGACTT CCAGCGCCGAGGAGCCGACGGGCAACAGGCGGTCATCACTCATCGGCGCCCCCGGCGGTGAGGGT GTAATTGGTGCAATATGAGGCCTGGTGTTTACCGATCACAATGTCAGCAAGCGGCGCGGTCAGCTCG ACACGCTGCACCCCTTCAACATGCAGGGCGGCATAAATGGCCGACTGGCGGACATCACGCCCGAGC CGGTGCTGCGCGGTGATATACGCCTCCAGTTTTGTCTTTGCGGCGGTGATGATGGGCTCCGCTTCGG GGCCAGGGTAAAAATACAGCGTGGCGGCAACCTGATACGGCACAATCTCAGCGGCCTGTACCGTGA CGCGATCCCCCACCGGGCGCACGTCTTCGGCGTTAAGGGCTTTCTCCACCACGGCCAGCAGGTCAGC ACCGGCGGAGCCGTCCCCCTCGCGGGCTAACACGGTGATTGTCACACAGGCCGGTGTCGGGCTGAC CACAGAGATATCGGCGACACGCCCGTCAGCACTGCGCCCGTGGTACTCGTAAGCGCCGACCGGCCC CGCCACGCTTAACCCTTCAAACGCCTGTTGCGCGCGTGAACGTAAATCCGCGTCGGATTCCAGCACC GCCGCCACAGGAGGCACGGCGGTATCATCCGCCGGGGTGATGGTCAGGCGCGCGGTGTTGCTGTTG GCGGCCATCACGTCAAGGTCATTACCGGCAGCACTCGCCAGCATGACGGCGCGGGCAGCTTCGTTA ACCCGCTGGCGCCAGATAACTTCGCGGTAGGCGTTTTCCTCAAGGAATTTTGTCAGCGGTTCGGACT CAAGCGAGATCGTACGGGCAACGGCGTCCTGTTGCTCCTCCGGGAAGAGGGAAACCAGCGTCGCTT TACGCTCGGCGAGGATACTTTCGTAATCCAGTGCCTCGACCACATCCGGGGCGGGGAGCTGGCTCA Attorney Docket No. PIV-00053 WO

[0305] AATCAACAATCGGCATGGTATCAACTCACGGGAAGGGTTAACGAAAGGGTTTCGCCGGTGCTGGCT ATCTCGCCGGTCAGGCTGACCACCATGCGGCCATCAAACTGACGCTCCGTTGTCACCGTGCTTAGCG TGATACGCGGCTCCCACnCAGCACCGCAAAATAACAGGCGACTTTGATTTGCAGCTCCAGCGCCGG GGTCTGCGGCTGGTCAATCATGGAGGACAACAGCGAGCCATATTCCCGGCGCATCACGCGCGAGCC GACCGGCGTGCGCAGAATGTCGGCCAGGCTCTGGCTGATGTGCTCCGCATCGGTGAGTGTGCGGCC ATCGCTGCGGCTCATGCCAAGATAACGGGCGGTCATAATGGCGCTCCTGTCGTGCCGCCGCTGTCGC CGGGGTGTTGGTGGGTGTGGAGGACTTTGCCATTAGACGACAGTGCGCCGCCGCTGTGCTCGATAT TGCCGCGCATCGTGCCGCCGCTCTGTACCTCCAGCGAACCGGTGATCAGCTTGTTGGTGCAGACCAC TTCCGGGGTATCGAGCGTGATGCGGGAGGAGGCTTTTACCATGACCACCGGCACCGTGACGGTAAC GGATTCCGATGCCGTCACGTCAGCGGTTTTAATCCCGCTGACCGTCAGCGCACTGGTTTGCGGCTCAT ATTCAAAAACCGCGCCGTCCGGGAAGGAAATATGCCAGGCATCCGCCGAGGCAGACGGGGCGGGG TTCTCATCGGAAAAAATACCCGGCAGCACAAAGGCGGTATCAAGCTCACCACCGACGGCCAGAATTA ACACCTGCTCGCCAACGGAGGGAGCCCACCACGAACGTGAGCGCCCGGCGCGTTGAGTGAGCCACT GGAGCCAGTCGGTAACAATGCCGCCGGTCTGCACACGGCAGCGCCCCGCACCGGTGTCAGTTTCGA CCACGACGCCGGTGCGTACCATATTGCGGATAGCGCGGGCAAGTTCGTTAATGTTTGCTAGTGTGTT CATACAACAGAGAATGCCGCCAAGGAAGGATAGCGGCAATAAAGGCGGGATGTACTTCTGTTAGCA CAACTTGTCTTAGTGCCTCTCTCTTATTATTTTATATACTGCGCTCAGGAGGTGATAAAGATTGTAATA TTGACCTTTTAGCAACTTATCCATATCATTTTTATTAAAGCGAGCTCGTTGTAAAGGATCTATATAATT ACCATCCCACTCGATGAATTGATGAACTGTATGCAGAGAAGCTATTGAGAGCGGTTCTTTATTTTTCT CACTGTAAGAAAGCATCACTTCGGCAGACTCCGCATAATAGTCAGAAGGGAAATTTTTAATAATCTTT CGAGTTTTCCACCGCCAGAATTGTTTCTCTTCCATATAGATTTCTTTGTGGGAGGACATCATAATTTTC TTCCCAAGCCTCTCAGAAATTTTCAAGAACCTTGCGGCGGTTATCCAAACCATTCGATCACATGGGGG ATAATCCTCACCTTGTTTAGGCTCAAGCATTTCATATGCTCTCTTCAGGTACTCTTTAGCATCTTTGGTG TAGTCTTCTGAAAGATGACGACGAGCATTATAGATAAGGCTTGTCATCGTTATAGAGGATGCAAGTA GCGCAAGATAAGATACTGCACCCTTAGCTTCATTAAAAAAAATGGAATTAGATAAAATAATAGCCAA TACGATCATGATAATAATGGAAAAATAAAACATTAAATTTACAGAGAAGAGAAATCGAAAACAACCG TACACCATCTTAATTAAACCAGTCACCACTGACACCACCGTCCCAACATTGAAATAACGCTTAATTCCC ATTGAAATAACCGCCTATAATTCAGAGAGATGATGAAGAACTAAATCATGAATAGTCTTAGCATCAC TGGAATCAAAAGCAAGCAATATTCGCTTAGAATAGTTGACTTCCTTACTGTTTTTACTGGGCCTGTCC TTTAATCCGAGCTGATGCACCCGCGCGATACGCTGCACTTTGCCGGTAAATTCCACCACTGCCGCACT GTCATTACCACTGGCTTTTAGGTAACGGTTCGTGCGCAGCTTTGCGAACATTTCCCGTTTAATCCGGC CTTITTTCCCGCGTATCAGCTGTGTTTTTCGCTTCGCGTAGGGCGCTCCGCTCGGGTCTTTCTGCGATT TGATGCGTTGTTGCTGGCGCTGGCGCAGTTTCTTCGCAATCTCGACAGTCAGACGGCGACGGCCTGC GGGAGACAGTGCCGCCAGCAACCCCGCGAGCTTGTCCTCAAATGGCTTAAGCTCATTCATGCCACTG ACTCACCAGATCATTACCGGCATACATCGCCAGCGGTCGGGTTACCGGCTCCGGCGGCGGTGGCTCC GGGGCAGTGGTTACATGCAGTGCGGCATCAACCTCTTTAACAATCGTTCGCTCGGTCAGTAACAGGC TGATACTGATATCGACGCTGCTGTCATTGTTGATGTCGGCATAAAAGGTAAAGCCTTTTTTCTGCCCC TCGCCGGTTGTCATGATGTCCGGCTGGTACTCACGCAGCCAGGCCATCACAGGCACGAGGATCAAAT CAATATCGCCGGTGTAGTCGGTCACCACCACGTTGAGGGTGTAGCGTnTTCAAATGACAGTGACGC GGCAAGCGTTGCGCCAATGCTCCCGCTGTCGATAAACAGTCGCAGCATTTCGGGGTTGGTGCGCAG CTCCGGCACCGCATTACTGAGGGCTTTGCGCAGGCTGTCGGGTTTCAGCATCGTTATCGTCCTGGCAT TTTTTGACGGCTTCAACCTGTAACGCGCAGCTTTCTAACGCACGCTCAAGGTTCCTGATATCGGCGCT CAAATCGCCGTTGGTGTGTGGATCACTTCCCGGCATCGGGCAAAGGCTGACTTTCGGGCAGCCGTTG TAAACAATCACCGGCACTGACGCAGGTGGGGCGCTTGTGCAACCGGCGCACAGCATCAGGTAAATC AGTGCTGTACCAGCGGCGAAAGGCTTCGTTTTCATTGAGCAACCTCGTTATGGTTTGTTCCCGCCTGG CTTCCCGCTCACCGGCGGCATTCAGCTTATGGCGCAAATCCACCTGCGCCCGCTCGTTGTTATCCGCC ACGGTACGGGCAACAATGAGCTGGTTTTTCAGCATGCCGATTGTTGTnTTTGTTCACTGGCGACACG GTTAGCCTTTTCCAGTGAGCGGGTAAGATTGCTGTTTrCATGACGTAACCAGAGCACCCCGGCGCAC GCCAGCACGACAGCGCCCAGAATCAACATGATTTTTGACACAGGCCGCCCTCCTCAATGCGCTTTCTG TACGATGAGCGCACCGCCCAGAAACCAGCAAGACAGAGCAAATAAAACAGCGCCGTAAAAATCGCC Attorney Docket No. PIV-00053 WO

[0306] CCGCCAGAGAGCAGACAAGGAAAGGTGAACCCCAGAATTAAAAGCGACCAGAATTTCCGCCAGGCT GGCGGTTGATGGAAAAAAATAAGGCATGCAATGAGCGCCGCATAAAAACCGCCTTTGCTCCCTTCAC GCCCATGTATCAGCCAGTACTCAGAAGCCGCCGCGCCCGTGAGGCTGGCAACGAAACACACCAGGC AGCAAAATAACGCCCATATCACAATGAAACTGACAGCAACACTTTGCGGGTGAATCAACCCGCACAG AAGCAACAGCGAGATAAACGCTTTTAAGGCCGGTAAAATTAAAGGCTTTTTCATTGGGTTACTCCTTT TTTGCAGTATGCCCGTTCCCGCGCGCGGCGGTTTTCGAGCCCGGTATTTTTAACGCCGTTGATGTAGA CCCAGCGCGACAGTTGATCGCATGCCTGTCCCCATTTTTTCTGGTTCAGGAAATAAACCAGCGTGGAC TTACACGCCGCGCCGGTGCCGACGTTAAAGGCGAAACTGACCACGGCGTCGTAAACCTGCGGCGGC ATGTCGACCGGCGCGCAGACCGCGAGACGGCGCTCAACCGTCAGCACGTCAGAAACCAGATTTTGC GCCGCTTCCCGTTCCGTAATATCCCGTTTCGGCGTGACCCCGGCAGTGTGGCCGATGCCTGATGTCCA TACGCCCGCGCTGCACTGGTAGGGGCGCAGACGGCACCCCTCAAGGTCTGCAATCAGGGCAAGACC GTCCTGCGAGGTATGGAGTAAACGAAAATCGGGCACCAGTGTCGCCAGCGCCAGCACGGCGGCCAC ACTGCAACGTTTAACGATrGATGACATTGCTCATTGCCTCCCCGGAAAGACTGCCCTGTCTGACCTGG AGCGCTAACAGGGCATAGCTTTTGCGCCGGTAATACCAGTTAACGCCGACCGTCAGCACCACGCCCA GCGCGCCAAAATACGCGGCGAAATCCTGCGGGGTCATCGCACCGAAGAAGGTGAGCGCAACGCTTA TCCAGTACGCCAGCGACGCGGTGATTTTTTCCATGCTCAGTCCCATAAATTCACCGTTTCCCGTGCCG CTGCGGTCTGTATCTCGGGCAGCTCCACCGCTGCGCCGTGGGGCAGTACCGCGCCCAGCTCAGCCAG CCCCGGATTAGCCGCGAGCACCGTTTCGAATACCCCCTCCGTGCGCCCGTAATACCGGGCGCAGATA ACATCGAGGGTGTCGCCCTGGAGCGCATACACCTTCATCAGATTTGCCCCACGATGCAGCGGGGTTT GTCCTGAATGCGCGCCACTGACCAGCGCATATCCCGCCACAGCTCATCAATCGTGGTATCAATGCTGT CGGCTTTTTTGTCGCCCTTGGCGCTCGCATCCACGCCGCGATAACGTTCGTACAGCGTGGCGGTTGCC ATCGCACACACCGCGCGCTCGTAATGAAAAACCCGCGTGCTTTCGCCGTCGATCTCATCCGACGGCA CATCGGCCAGGCGCGCAAAACCCGCCTTAATCTGGCTGTCGCGGTACTCGTAAAGCTCCGCGTTCGT CTCGGCAATGCCGGTTTTAATGGCGTTACGCAGCCGTGCCGGGGCGATGGTCTGCTCAAGGCGCATC AGTTCGCGCACGCGCTTCGGGTCGATGTCGGGAAAGAAAAACGTATTTTTAATGACCGGCTCGTCGC CCGCTGGCTGCGGAATGACAACCGTGTTTCCCGGTTCAGCCGCCTCGTTTTTGTTAATAATCACTGTC GTCATGACTACCTCTGAAAGGGTGGGCGGTGGACGCCGGTCGCTGATAAGGCGAAACACCCTCATT AACCGGCGTGCCGCCCTGGCGCGGGGGCGCGTTCTTTTAACCGGCGACTTTGCGGGGGCGACCCCG CCCGCGTTTCACCGGTGTGACTGTTTTTTTCTGCTGTGCGGCTTTCGCGGGCTTTTCGGGCGCGGGCG TCGCGGCAACAGGTTTCGGATTCAGCTCGCGCGTAAGGCGCTCAATGTCTTTTCTTACCCCGGCGTTG CGGTCGAGTTGTATCGCGCGTTGCAGGTGCGTCATAGCATCAGCGGGCTGACCGGCATCACGCAGC GTCAGCCCGGTGACTTTATGCAGACGGGCGCGCACCTCATCGGGCATGTCGGCTTTCGCTGTCATGC TGATGACATCGAGAAGGATCGCCGCATCGACCGGCTGACCGGCATCACGGGCGCGCAGGGCGGCA AGCGCCACCTCTTCGGCCAGCATGTAAGGCGCGGTGCGGTGATGGTTTTCCGGCATCGACAGGTTGT AGCGCAGTGCATAGCGGGCAATCTCCAGCGCGCCGGGAATGTCACCGGCATCGAGACGCCACAGCA TCACCGTCATCAGAATGGCGTCCTGCGTGCCAGTGCCGTTCTCCAGCACGCCGGTTACCCAGGGCAG ATAAAACGGCAACAGTTCGCGCTTTTTCTCTGCCTTGCGCTCTTTCGAATGGATCTCTrTTAACGTCCT GCGGTCTGCGGCCAGCTTTACAAGCATCTGCTCATAAGGGGTGGCATGGCGCAGCGGGTTATCTTCC CGCTGCGCTGCCTTCATGGCCGAGACCCGCATCGCGTGACGCTGTGCGGGGCTCGCCATCGGTTACG CCCCCTCGCCGGTGCTGGCCGGGGTGGCAGTGGAGGCGGTTGCCAGTGCTTTTATGGCCTCAGCCA GTTCCGCCGCGAACATTTTCGCGACATCCGCTTCCGGGGTGTCACCTTCGGCCTGTTCAAGGATCTCG ATGTnTCAATCAGGCATCCGGCTTCGTAGTCTTCAATCACAAAATCGACTTTCACCTGCTCGTAGTTT TCAACCTGGTCGAGCTTCGGATTTTCGATGATGTGGCGGCGGTGTCCGTCTTCGTAGAGGTAAATCG AAATGTTATCCAGCGTGGTGATAAAAATGCTGTTTGCCGGGAAGAACGGCGCGCGCACGGCCTGTA ACTGGCCGATGGTTTTCTGGCTGATAATCAGCTCACCGGCCAGTTGCTCACTGTTGGGCTGGAATTTA TTGATCATCGGGAAATATTTGTCGGTCAGAATACGGCGACCACAAATCACCACCATTTCCGGGTTTTC GCGGTGAACCTCCTCGATCAGCGACTCGTGCGCATCCATGACCAGCGCGTCGAGGTTTTTGTAAACG CCGTTTTTGCCGACTGTGATnTCTCGGAAATCACTTTGCCGCTTTCGTCGACAACCTTGCTCATGACG CGCTCTGGTGCGTCGTTGCGGTATTTCTGCAACCAGCCGATTGCGACATCCTGCAACAGCGGATTTG CCTTGCGGTCAGAGGTTGCCGCACGGCTCACACCGTTAAAGCCGATGGTGATGTAATCCAGCGCCTG Attorney Docket No. PIV-00053 WO

[0307] ACGCTTGATGATCGCGTTACGGATACGAATCTGAAAATCCTGAAAACGCGCCCACAAATCGAGTTTG TTATAGCTCAGGTGATAATCGAAGTTCACCGGGTGGCAGAAATAGCGGTAAGCGTCCATTTTCGAAA AATCGGCAGTCTTACGCTCGTCACCATTTGCCGTGTCGGCAGTGCTGGCAATCGTGCCGTTAACGTC GATGCCGACTTTTTCCTCGGTCAGCTCGCGCACCACGACCATATTAATCTGCTGGAGGAAGGAAGAA GACTGCTGAATTTTGTCAAACAGCGTCTGAGTTACCGACGGCTCAACGCTGAATTTTTTACTCAGGTC TTCGACGCTTACGCCGTTCAGCTCTGCAAGGCGGCTCATGTACGCATTAAATTTAAAACGGGTTTCTT TACGCATTGTCTCTGTATTCCTGTTCTTAAAAAGGGAGGCCGGTTAGCAGTCGGTCAGGGTTGCACC CGCGCCATCGCCGCCGGTGCTCAGTTTCCGGCGCGGCTGTGTCTGGCTGGGGGTTTTGTCCAGGGTG GCGGTGATCGTGCTGAACTGCTCGCTGTTTTTGGCGGCCTCGTCGGTCACGCTTTGTTTAAGCGCGG CGAGCTCCGCTTCCAGCGAGGTAAAACGCGCACCGGCGCTGTCTTCGCTGGTCTGCACGCGCTCGGC GATAGTCGTCACCGCTTCGTGCACGTCGTTAAAACGGGCGTCATCGCTGGTCTGTTTGCGGCTGAAA ATCGCCTTTACGGAATCGCTCAGCCTGTTCAGCAGGGTTTCGGGCTGGTCTTCGAATTCCAGCTCGGC GAGGGTGGCAACAGAGAAAACATTCTCCGGGCTGGATTTGAAACGCTGAAGCGGGTTGTGTTTCAC AGTGCGGCAGAATTCCAGATATTCAGTACCGAGGCTTGCCGGGTCATCAGTGACCGCCAGGCCAACC AGATAACACTTGCCGCTGTTGGCAAAGTTCGGCTGAATTTCCATTGAGGTGTAAACCTTCTGGCCTTT CGCCACCATATCGACCAGCGTGTCGAGCGGGGCAATTTTGCCGTACAGCGCCAGCTTGCCATTCAGA GCGGAATCATCTTCGATTTTCTCCGCTTTCAGTTCGACCACATCGCCATAACGGGCGAAAGCGCCATC CGGCAACAGGCCGCGCAGGTGTTCGAGGTTGATACGACAGCCATAGACACGCGGGTCAAATGAGTC GGCCATTTCCTGAATATCCGACGCGCTGATAATGCGCCCGTCGCAGGTGTCGCCCTCGACGCCGATA CGGAAGAATTTTGATACTTTTTTTGCCATCGTCAGGAGTCCTGATTGTGTGAGAAAGAATCACGTTTT GTCAGGGGGTAGTTTCCCGGCTCGTCCGTTGGTTCGCCATCAGTCACGGATGGCTTGCCCGCGACAC ATCAGCACCTTAGCGAATCGCCGCTCCCGCTTAAGTAGCCTTGCCGTGTACCAACAACGGCGAGGCA GGCATGACCATCACCACCGACACGACACTCTTAAACGACCCGCGACGACAGGCGGCGCTCCTGTACT GGCAGGGGTTTTCCGTGCCGCAAATCGCGGAAATGTTGCAGACCAAACGCCCCACGGTTCAGAGCT GGAAACAGCGCGACGGATGGGACGACACTGCGCCCTTAAACCGCGTCGAAAATACGTTAGAGGCGC GGCTCATCCAGCTTTACGCCAAGCCGGACTTGACCGCACACGACTTCAAAGTCGCTGATTTTCTGGCG CGCCAGATGGAACGCTTTGCCCGCATTAACCGCTATGGCCAGACGGGCAATGAGGCGGATCTCAATC CGAACGTGGCCAACCGCAATAAAGGGGATCGCAAAAAGCCGAAAAAGAATTTTTTCAGCGAGGAG GCGGTCGAAAAGCTGGAGGAAATmCTTCGACCAGTCTTTCGAATATCAGCTCGGATGGCATAAAG CGGGGCTTGAGCACCGCATCAGGCACATCCTCAAATCCCGCCAGATTGGCGCAACGTTTTATTTCGCC CGCGAGGCGCTGTTACGCGCCCTGAAAACCGGCCATAACCAGATATTTTTATCGGCGTCCAAAACAC AGGCGTATGTGTTCCGCAAATACATCATCGCCTTTGCGCGCCTGGTCGATGTCGACCTGACCGGCGA CCCGATTGTCATCGGCAATAACGGCGCGGAGCTGTTATTTCTCGGCACCAACAGCAACACCGCGCAG AGCCATAATGGCGACCTGTATGTCGACGAAATTTTCTGGATCCCCAACTTCCAGAAACTGCGCAAAG TGGCGTCGGGCATGGCGTCGCAAAGCCATCTGCGCACGACCTATTTTTCGACCCCTTCCTCGCTCGG GCATGGTGCCTATCCGTTCTGGTCTGGCGACCTGTTTAACAAGGGGCGCGCCAGCGCCAGCGAACG GGTCAGCATTGATATCAGCCATGCGGCATTAGCGGGGGGCGTGGCGTGTCCTGATGGTCAGTGGCG GCAGATTGTCACGATTGAGGATGCGCTCGCCGGGGGCTGCACACTGTTTGATATCGACGCGCTCCGC ATGGAGAACAGCGCAGACGACTTCCGCAACCTGTTTATGTGCGAGTTTGTTGACGACAAAGCGTCGG TATTCCCGTTTGAAGAACTGCAACGATGCATGGTCGACAGCATGGAAGAGTGGGAGGACTTCGCCC CGTTCGCCGACCGGCCATTTGGTCACCGCGTGGTGTGGATTGGTTACGACCCGTCGCACCAGGGCGA CAGCGCCGGGTGCGTGGTTATCGCGCCGCCGGTGGTrGCCGGGGGCAAATTCCGCATTCTGGAGCG CCACCAGTGGAAGGGGATGGACTTCGCCACACAGGCCGAGTCTATCCGCGCACTCACGCAAAAATA CAACGTGGAATACATCGGCATCGACTCAACCGGGCTCGGTCAGGGCGTGTTTCAGCTTGTGCGCTCG TTTTACCCGGCTGCGCGTGACATCCGCTACACGCCGGAAATGAAAACCGCAATGGTGCTCAAGGCCA AAGACACCATCACGCGCGGTTGCCTGGAATACGACGTGAGCGCGACCGACATCACGCAGTCGTTTAT GTCCATCCGTAAAACCATGACCAGCAGCGGGCGCAGCGCCACCTATGAGGCCAGCCGCACCGAGGA AGCCAGCCACGCGGATCTCGCCTGGGCAACCATGCACGTACTGATTAACGAACCGCTGACCGCCGCC ACCGGCCAGCAATCCTCTTCCATTCTGGACTTTAACTGATGAGCAAAAACAAAAAACGAAAATTCACC CAGGCGGCGCGCCAGCATACCGCCGCACCAGCGCAGAGTATGGAGGCGTTTACCTTCGGCGAGCCG Attorney Docket No. PIV-00053 WO

[0308] GTGCCGGTGCTCGATAAACGCGAAATCCTGGATTATGTGGAGTGCATCGATAACGGCCAGTGGTAC GAGCCGCCGATCAGTTTTTCCGGACTGGCAAAAAGCATGCGCGCCGCCGTTCACCACAGCTCGCCGA TGTATGTTAAGCGCAATATTCTGGTATCGACGTTTATCCCGCATGCCCGCCTGTCACGGCAGGATTTC AGCCGGTTCGCCCTGGATTACATCGTTTTCGGCAACGCGTTTATGGAGACGCGGTTAAGCCTGTCCG GCAGACCGGTTAAATACGAAACCTCACCGGCGAAATATACCCGGCGCGGCGTGGAGGACGACACCT ACTGGTTTATTCAGAACTTCACGCAGCCGCACCAGTTTGCGCCCGGGTCGGTTTTCCACCTGCTGGAG CCGGACATCAATCAGGAGCTTTACGGGATGCCGGAATATCTCAGCGCGCTTAATTCGGCCTGGCTGA ACGAGTCGGCGACGCTGTTTCGACGCAAGTATTACCAGAACGGGGCGCATGCGGGTTACATCATGT ACGTAACCGACGCGGCGCAAAGCAGCACGGATGTTGAAGCACTGCGCAAGGCGATGCGGGATTCG AAAGGGCTCGGCAATTTTAAAAACCTGTTTTTCTACGCGCCGAACGGAAAAGCCGACGGCATTAAGA TCGTGCCTCTGTCGGAAGTCGCCACCAAAGACGACTTTTTTAACATCAAAAAAGTCAGCGCCGGTGA TCTGCTCGACGCGCACCGCGTGCCGTTCCAGTTGATGGGAGCCAAACCCGAGACAGCGGGCTCTATT GGTGACGTTGAGAAGGTGGCAAAGGTGTTTGTGCGAAACGAACTGAAGCCTTTGCAGTCGCGCTTT CTGGAGCTCAACGAGTGGGCGGGCGACGAGATCATACGGTTTGAAAAATACAGCCTTGAGGACGAC AACGCGTAAACCATCACAGCCGCCTGCGGGCGGCTTTTTCATGCCCCCGCCCTGACAGCCTCAGAGA CGCCACACGCCGCGCAACACTCCTGACATGCTCACCCATGCCTTACGCCGAGACAACGCGACAGCGG CGCGCTCAGGCGCGAAAAAAAAATAAATACCCGCCCCAGCGCGCAATGCTTTCCCCGCCACGCCTGC CCGCTTTTCGGATCGGTTTTAATGCAGITGCACGTTAAAACCAAAGCCCATCAGTTTTGATGGGCTAG CGTAAAAACGTCCAACAGAAGTGAATGCAGATTCATGCATATAAACATGCGTGGCCGTAGGATAAA AACTATGCAGCCCTGCCTAAATTCACATTAGGAATTTTTAAGGCGTCAGAAAAAATCATTACCTCTTC ACCGACCCGTTTACTTTGAGCCGTATAACTCAGTGAGTATTCTGTCTGTCTAAATTGTGAATATATTTG TTTTATCTCACTACTGTTATCATAAGAGACAAGCCAATTTTTAATTCCTGAGTTTCTTAAAGAATCCAT GACATTGACATGGTCTTCATGTTCGTAATAATTCCTATATAGATCTTGCCCCTTAACATAATAAGGCG GATCCAAATACACCAACAACTTATCATCAGACATCTTGAAATTATTAATATTTAAAAGCAGCTCAGCA GCATCAAGATTAGTAACCTTAATTCTATTTTTATATCCGGATATTTTATTAACTCTAGAAATTAGATCG CTTTTATTAAAGCGAACATCCATTTTCCATTCACCCAATTGCGACTTGCCACCAATAACACCGGCTTTT AAAATTCCAGACCTATTAGTTCTGTTTAAGAAAAAAGCTGCAAAACCTAATTGCAATGAGGGTATTG GATTGAATGAATTTATTATTTCTCTTTGTTCATACCACGTATCCATAGTGATATCGCATTTTTCAATCAT TGAACAAAGTGCATCGGTTTCATTTATAACAGAGTGCCAAAAACTAAACACAGCCGGATCGGCATCA TTGATGAGGATATTTCTTACATATTCATTAACAACCAAATCTAAGGCAATAGCAGCCCCGCCAGCATA GGGTTCAACATAACAACCATCAGTGATGAGGTTTTCCTCAAGCAGAGATTTTATATAATAAGACAACT TTCCCTTACCCCCGGGGTATCTCAGCGGCGTATAAAACCTCATACAATACAACCTCTTATAAAAACTTA TCTATCAATAATGTTAGCCTAACACAATCCCTGTGAATTTTCCATTAGATAACATATAGTTATGATGAG ACACATTCCCAAATAGCCTCAATCAAAGGCTGAAAATTATCCCATTCTGTATTTACAAATrCTTTCGTT GGGATCAAATTn'CATTATGAACATACTGTTGTAAAGAGGAGCTAGAATTTGTTGTTTGTn'CGAGTA AGTTTGTATGGCAGTTTTCTTCGGCCCATTAATTAGTTTTTTCTCAAATAAATCCGTGGAAACCATTAC AACCTTATCATGTAAACCCGGTGTTTTTTGCGGATTCTTTCGATCATGATAAATAAGATTATTACTATC AATATATACATTCAAAGAAAGTTCAATGAAAACCCTTAGCATTACTGCCAATGAATnTGATGCTCAT CATGAGTCATACGTTTCAGTTCTGAAAAAATACGATGACATCTTTTATGCTCTTTGCCGAACTTCAGTG ATAGCCCTGTAGGTATCATATGGTTCCGGTTAACGCTTGCCGGGTTTATCCCTTTTTTTGCCTTGTTTT TATCATCATCATCGGTGCCAAAATAATCCACGCCACCGGGATTATTTTCACCACCACCCACAGGCGTT CCTTGGCCACCTTTAGTATCTTCACCGCCCTTTTGACCACCCTGCGCGTTACCGTCTCCATCAACGGAA TTTTTAGGCGGCTCTAATAAACGCCATTCTTTTTCTAAAGCCGGAGCAAAGGATGTTAATTGTAACTC TGAAATAAAGTTACTACGATCTAATTTGAAACGAATACGGTTAACAGTAAACTCCGTTCTTCCACTCT CGTCAGTTAAAATCATCACATCTAATACTTTTTTAAATTCGCTAATGAAACGTTTTAACGGCTGTGAGC AAAAAAGGACACCATCGACACCATTAAGATTCAAACTTTTCCTAAAATTCGGGTCGCCAACCAAGCG ATCTAGGTTAGTTATTCGAACTCTATTTTTTTCTTTAATAATATTGTCAAATTCATCTGGGAAAAGATC AATAAATGAATAAAGCTGGTTAGCAAAGGACTCAGTACCGTrTCTAGCCATAAATCTTAGCTGTTCAG GTGTTGTCCATCCGACACGACCTGCTCCTTCATTTTGACCAGTATGTTTTAAATTAATCCAATGATTAT ACCCCTCATCATCATACACTACGCAATCAACACTTAATATATTGAATTTCCTTGCCTTTTTAATCTTCTC Attorney Docket No. PIV-00053 WO

[0309] GAATGTTGATTCTACTGTCTTATTTTGCGCTAGTGATGGGTTTTCAATTAACTTCAGAGCTGTAATTCT TCTATTACCTTCCTTAACAATAAACCCTTTTTCCTCATCTTCATTTTCATTTTCAACAACTATCATCCGTT CAGAAGGGTCTATTCCGTTATTCGCAATGTCTCTTGCTAATTTTATTAACTTTGCCCCCTGGCTCTCAA CCATAATTTTTATTGCTTCTCGTTGTCCATCCGCATTTTCACTAAAACGAGAGTTTTGCACATCTAGCAT TAAGTCATTTATATTGATGCTTTTATATTGATACATTCTAACGTCCTTATTGCTGTGAAGTTTTTACGCT ATTGAGAACAATGTTAAGTGATTATCTACCGAAGTGGTGATCATCTCAGTTTACGTAAAAATCGTATA GTAACAAGCACAAACAACCACGAATTACAGCATAAATGAATGTACCGTTGTTATCCCAGATATAAATT ATTCATCGCGATACGAGCTTATTTAATATACAAAGCTAACTATAAGCAATGCTAATTAATTACTATACT GCCCTTCATTTTTTAATGATTTCAATTGAAGAATCCATAGAAAAAACTTTTATTTTCAATAAATTATCCC GTGCAATTTCAGAAATTAAATCGAGTGCTATTTCTCGATCCCGTTCATGGCACGTTCCTTCGGTAGTC AGCCGTGCGATCAACTCAACTCGCTCAAGTATGACCAGTTCTTTTAATTCGTTTCTCACAAAACTTCCC TCACATGAGATACTGTATAAACATACAGTATAATGATTGTTTGCACAAAGTGAAATAAAAAAATCAA GCATAACCCTTTGTATCTTTGAGATATAAATTTGAACGTCTTCTCAGTTTTCTTTCACCATTCCAGACAA AACGGCAACACAGGCAAGGATTTCCTTAGCCTTTGCAAAAAATGGTGGGGCGGTAGAAAAAACCTC TCCTGCTTCGGATCCCCGACACCATTTTCCGTTAAAACAGCTTTTGCCATGCGCCATCAAGTGCAGAG CTTCGCCACGGCTTATAGTGATGCCGGTTAAAAGCTGTATTTCGTTAATCGTCCTAGATATCGCGCCG TCCTGCGCAGGCGTTCCGTGGATGAAATTACGGCGCTCGGTCGGTTTTTTTTCCCTTAACCGGTTAGT TAGCTGCCGTCGCTGATATCGATTTAATGGTTTTGATAAATCCAGCTCCGGCGGATCGCTTTCGCTTC CCGTACAGTTATTGACAGAACTCCGAGAGGGCGCGGTCGCGCCCTTAAGGTCAACGGCCAAATCAA CGGCACGCTTCGGAACAATTTTCCATTGCGTGAGGCGGGTTAAAATTGGTGAGCCAGTACCGACAGC AGCATCGTATACGCCACGAATGCAGACGGTTTCCTCGCCGTACTGATTAAACTCGGATCGTGCTTCGT AGAGCGTGCGAACCTGCAAATCATCGCGACGGACAAACGCGCCGCCCTGGGCGTTAACGTAACCCG CCCAATCACCGGCATCAGCCGCATCATGCACCGCAGCAAATTCAACGCTAAGACCGTGCGCGGTTTC GGTGTCGGCCATGCGGCGTAATTCGCGGTAAACCGTCACCGGCGCACCGCCGATAAACTGGAATTG CCGGATATGCCAGCGCGCCGCCCAGGCAGAAACGGCGGGCGCGGTATCTTTCAACAGCTCGCCGCT TTCGTCGTCTGTTTCGCCATCGAGCGCATAGCCATCGATATTCTTTGAGATGTATTTTGCGATATAGCC CGTTGCACTCCCCrnTCCGGGTCGATCGCTTCAGCGTGGAATCTGGCCTTCCTGGCCTTATAGCTGCT GAGCTCGTTGCCATCCTCCTGCCACGCATAATCCCGGATGACAGTGCGCACACGCTCGGCATGTTCC GGCAGCATAAACATCAGCATATGCCAGTGCGGGGTAGCATCGTGGTGAGGTTCTGCAACGCGGATC CCAAAGATGCGGATATCTTCCCGGTGGAGCTTGGCGCGAATGTGCGCCCAGAGTGAGGTTAAATAG ATTTGCGTGTCGGACGGGTTTGCCCCGTTCCATTTGTGATTACGGTAGCCCGCTTTAGTrGTGGCATG ATATTTCGACGGTGCGGTCAGCGTATAAAATTCACCGACATAACCAAGCTCATTACAGATATTTTCGA ACCCACGAATGCGGGTCATCAGCTCGCAGCGGCGAATGGCCGGGTTAGCGACCGAGCCGTCGTATT TTTCAATCAGGCTGATGCGGTTGCCGTCCTCGTCTTCAAGCTCCAGCCCTTTAAGAAATTCACGCGTG CGCCTCTTTTGCTCCCGCCACTCGGCTACACATGTTTTGCTGGCGTAGGCATGCTTTTTCTTGCTGACG TTACCAACAGCGATTTGCAGATGCTCACGCCATGCGGCAGCAATACGACGCAACCGCCCGCGCCACC ATGTTTCATTGAACATGCGCATAACTGCCGGGGCGATATCCTGCTCACTAGCGTATmTTTGnACG CGCTCCCAATGGGGCGGGGTCACGTTAAATTGCAGGGCAATCAGCCCGGCGCGCATATACCACGAA TGCAGTGTTTTCAGTTCACCGGCGCTGGCATCGTCTATATCTGCAAGCTCCGCGCGGATGAAATTAGC GATATCAGCGGCCAGCAGTTCAATATCTGCCTTTGCCATATCAGGGAGCCGGTTATAACGGGCGACC ATATTCACGAGGCGTGAGGCGATGTATTGCAGCATTTCCGTATCGAAATGACCGGAAAAAACAGCA GCCGAAACATCGCTTTTAACGCCAGTGGCCTGGTATTTTTTTGTGATCAGCTCAAGGCGCGGCAATG CTTTTTTGCAAAAGCTGATTAAAAAAGCATTGGCTCGCTGGCTGTCGTGATGTTGCTCCAGCGCGGC GGCGGTTCGGTATACGTCGTAGCGTACACATTCCGGCTGGAGAGAAAGCGCTTTGCGCGCATGCAG CAACGCCGCAATCATACGATCACGGCGGTGCTGTTCCGAGTAGGTCAGATATGGGCTGGCTATTGCT GAGCGAGGAGCATTCCAAGCGTAAGCATAAGAAATTGCCACTTAAACGCCCCGGTAATGTCTGGATT TGAGCTCTGTTATTTGCTGGCAGGTCACACAAAGCGACACGCCATCAATTGCCCTGCGGCGCGCCTC CGGGATTGGTGAGTCGCACTCTTCGCAGGTAAAACGTGAAGGTGTAAAAACGCGGCTGCGCGCGTT ATTGATATGGCGCTCACGCTCTTCCTGCTCGCGTTGCTGGATGTGATCCATTGCGTCGGCCATTAGTG CAGCTCCTGTGATTCGTTTTCGTAGCGAACAGCCTCGCGGCGCAGAAGCTCGGCGGCTTCGACTCCG Attorney Docket No. PIV-00053 WO

[0310] TTCAGCCCTTCTCTTGCGATATGAATAGCCAGAGCCTCAAGACGAATGGAAACAGCAAGAGCACGAT CACGACGCTCCTCGTTTTTAGCTTTATTCAGCATGCTGACCAGTACATCGTTATCAGCTTCAAAATTGC GGGTTTCGATATTTTGCATAACGCTTTCTCCTGAATTTAGGCAAAAGAATGCCCGGCGGGTTGACGC CTTTTAATTACGGTTTGTGTTAATTCGGCATGGTTAGCCGTTTGGGAAATAAGCTCACGACCGCGCGA AAGTGATTCATTGCTGAAATAAGCGCTTTTTTCTCGTCAGTAGTCAGCTCACTTAATTTGAGCTCGTGC CGAGCTGCCGGTATTTTTGCCAGAAAGAAAATCGCTGCCAGCGCGCGCTCGTTGTCTTCATGCTGCG GGTCGCGTGTATCTCGCATATTTGCGACGAATTGCTCGACCTCTTTCCAGCCATCACCCCAGAACCGC CCGCGAATCTCAGCGATATGATTCAGGCCAGCCATACGGGCGCCAGCTTTCAGCGGAACAGTTGCAG AAACAGCTTCGATAGCCATGAGCCCTCCTGTTTTTTGGTAGAAAGACCAGCCAGCAAGTCAGCCTGA GAAAAACACGGATGCCAGCGCGTACCGTCTTTACCTGCGATCCAGCCGTGGCCGTAATGCATGGCTG GGCTTTGCTTAACAAGTAGCGATGCGAAAGAAGGTTCGTTATTCAACATACTCACCTCACATCAGCCC AAAAGATGCGCCGAGGCCGCTAACGGTATCGACGACACTGGACATTGCCGGGTTAGCCTGGAGGCG TGCTTGTAGTGCCAGAGCTGTCAACGAAAGCATGCGTATACCAGAATTCACACTCTCGACCATACTGT GCTTCCTTGCTGGGGTAAGACGTTCAGTGGAAATAGCACTGTCAGCCAGATCGCCAAGCTCAGCCAT TGCGCGCATGACATAGATTTGTAATTTCTCTTTTGCCAGCTCGTTGACAGGTACACAGGGTAAGCAGT GAAGCATTGCCAGAAAGCCGTCTACGAGCGCCGAGTCTTCAGTAAGATCGGTAAGTAGCCAAATCTC CGTCGGTGTGAGGTGGTGAGGCTGTTCGGGGTTGAGCTTGTTGCGCAGAGTCTGGACATTCATCCCC GCACGCTCAGCCAGCTTCGCCATATTGTGACGCTGTGCGAAAACCCGGCACGCTTCGTCATAGTGGG GATGTTTGGAAACCTGAAAATCAAACATGCTGCATCCTTACAATTCACATAAAGTGAATTAAGCGCCT ATGACAAGCTGAAAACGGGAATGCCCCAACGCCTTACGCAACTGTTCTTCTTTCCAGCGTGCGTAGT AGATGCGAATAGGGCCACCGGCTTTCTTGCAGCCTTTACGGATGGTGCGCGGTTCGATTGGTACACA AGGGTTGTCGCCGGTTGTCCAGCGGTAAGCGGTTCGCTCAGAAACACCCTCAAGCTCTGCGAACTGT TGCAGGGTAACGATAGGTGCAGGCACTTTGATGATTGCGATTTCAGAAGCCATGTTGCATGATTCCC CTTTTGCTAAACATTGCAATTAATAGCCATCTGTTTGCCAACATTCGCCATTAATTGCCTAGGTATAGG CTTAACATAACTCCCAAAATGGAGTTTGTAAATAGGTTAAAGCTACATGAGAATTGAAGGTCTTGGG TTAAACAACGAAGAAGTGTTAGATAGGATTTGCGAGGCTTATGGTTTTTCGCAGAAAATTCAATTAG CTAGACATTTCGAAATTGCATCCAGCTCTCTTGCTAACAGGTACAGCCGCGATTCTATTTCTTATGATT TTATTGTGCATTGCGCCCTGGAAACTGGTGCAAATCTCGCGTGGCTGCTCACTGGCAAGGGATCACC AGCAACCGGAAACGCGAAAAACGATACCCAAATCGTGGAAAAATTCACATTAAGTGAAGAATCATT AGTTGGTGATGGTGATTTGAGCATTGCCGGTAAGTTCTrTAGCAAACCGCTAACAAACCCTATAGCT GTCTACTCTGACGGAAAACTTCATTTTATCGAACGAGATGCCTCTCTTTCTGACGGGGAATGGCTTGT CGATATTGAAGGCGCTATTAGCATTCGAGAACTAACGAAGCTGCCCGGCAGAAAGCTACATGTTGCT GGCGGTAAGGTTCCCTTTGAATGTGGATTTGACGACATTAAAGCGTTAGGTCGCGTGATGGGTGTAT ATAGCGAGGTTAATTAATGACTGTCCGTAAAAATCCTGCTGGCGGCTGGATTTGCGAACTCTATCCA AACGGGGCAAAAGGCAAACGCATTAGAAAGAAATTCGCCACCAAAGGTGAGGCATTGGCCTTTGAG CAATACACAGTACAAAACCCGTGGCAGGAAGAAAAAGAAGACAGGCGTACGTTAAAGGATCTGATT GACGCGTGGTTTAGCGCTCACGGCATAACTCTGAAAGACGGTCTCAAACGCCAGCTCGCGATGCATC ATGCTTTCGAGTGTATGGGGGAACCGCTCGCGCGCGATTTTGATGCGCAAATATTTTCCCGCTACCG AGAAAAACGGTTAAAAGGTGAGTATGCCCGTTCAAACAGGGTGAAAGAGGTATCGCCTCGCACGCT TAATCTTGAACTGGCCTACTTCCGGGCGGTATTCAATGAGCTAAACCGCCTCGGAGAATGGAAGGGT GAGAACCCGCTGAAAAATATGCGCCCATTCCGCACAGAAGAAATGGAAATGGCCTGGCTAACTCAC GACCAAATTTCGCAACTGCTCGGAGAGTGCAAAAGGCATGACCACCCTGATTTAGAAACAGTGGTAA GAATCTGTCTCGCCACTGGCGCACGCTGGTCTGAGGCCGAGAGTCTGAGAAAAAGCCAGCTCGCGA AATACAAAATCACATACACCAACACGAAAGGCAGAAAAAACCGCTCTGTCCCAATCAGCAAAGAGCT CTATGAGTCTCTGCCTGATGATAAAAAAGGCCGGTTATTTAGTGATTGTTATGGGGCGTTCCGGTCA GCTCTGGAAAGAACAGGCATCGAACTACCGGCAGGGCAACTTACCCATGTTTTACGCCACACCTTCG CCAGTCACTTTATGATGAATGGTGGTAATATTCTGGTCTTGCAGCGAGTGCTCGGCCACACCGACAT AAAAATGACAATGCGATATGCGCATTTCGCGCCAGATCATTTAGAAGATGCAGTCAAATTAAATCCTT TAAGCTTTAAAACTAACAATTAAGAAAGGAGATTGAAAATGTCGACAAAAAAATACGAGCACATTAA GGAGTATTCTTTTCATGGAGAGTTTrGCCAATGTCCAGATGATAGCAAAGGAAGATTCTCCGCCAAG Attorney Docket No. PIV-00053 WO

[0311] ATAGAATACTCATCATATCATGGTTTAATTTTAGACTATTGTATTTCTGACAGTGATGGACCCGATAA GTGTGAAAGGCTCTATGGTTATTTGAACACTGGCGAGCAATGTACGTTAATTGGACCCTTTGACTTTT CACAAGGTGGCTTTCATCTTGGAAAAGGATTTACCAGAACTGGTCGGCATGGATTCGCTATCATTTTA TTCAATGGCTTTTATGATGAAAATCATAAAATAGAATATTGTGATCTATCTCTACATGGGTTGCAGGA ATTCATTCATCCTCAAGGTTTTATCACCCAACTAAAACACAAAAATGCCCCAATTTTTTCTGCGTCCGC AGAAGATTGGAAGATTGAACTCATTAATAATGCTACATTTAGCGTAGTCGGCGACGGGTTGTTAAAT ATTATATATTGTCAAGATGCAGATGCGCTCGCGAAGCTATCCGATGAATTTTTAAAAATAAAAGAGTT ACACCCATCAGCTCGTTTTTCAATTAGAAAAGATCTAACTTTCTATTTTAGATTTAAAAATCATAACTCT AAAAATATAAAAGAACACATGCTTAATATTTGGAAGATATCAGGGCTAATTTCCATATTAACCGACAA ACCTACCTTACCAGATGAATTGTATATAAAATTTGAAGGTAGTAATACCAGAACACCATGCTTATTAA CAACTAGATTTGAACAAAGAACAATAGATCTTGCACTGAAAAAATTCGATCATCGTTCTTTACCAATA AACTGGAAAAGCATAGATATCGAAAAAGCTTTCGAGAAATGGTTTGAGATTTCGGACAGATATATTC CTTTAACTATAACCTATCAGTATGAAACAGGCTACAGGACATTACACCAAGCACATTCTGATATTATC CTTTTCGCAACTCAAATCGAAGCAATAAACTCCACGCTAGGCGGAGAGAAAAGGGAGAAGTATATT AAACCGATTGATGAATATGCATCAACCTTTCTCAGGCAAAAAATGAACAATTTTTTTATTAGATTTAAT AATAACTCTTTAGGGGCAAATATTGCAACGCTTAGAAATGAATTAGCACATGTTGATCGCGATAAAG AATTGATGACACAAATGACACTAGATGAATACATCAGGATTGGTATGTGTCTAAGGTTAATCATCAC CTCTCACTTGCTGTCAAATTTGGGCATCGAAAAAGAGCATATACAACGATACCAAAACCGAGTTGCC CAAGATTGA

[0312] Prophage P-5812-03 (SEQ ID NO: 14) TTATGCAACTTCTGCCGCCATTTTGCCGCCATTAAAATGCGCCAGCGGGTTCAGGGTTACCGCTTCCT CCAGATGATCGGGCGCGAAGTGGGCGTAACGCAGGGTTTCACGGATATTGGCGTGACCCAGAATAC GCTGCAGTACCAGGATATTGCCGCCATTGGTCATGAAATGCGCGGCAAAGGTATGGCGCAACACAT GGGTTTTCTGCCCCTCGGCCAGTGCGATATCGGTGGACGCCAGCATTTTGCTGAACGCCGGATAGCA CGGCTGGAACATCTGCCCCTGCAATGGCGCAAGCTCGTCATACAACCACTGCGGAATGGGCACGGT ACGGTTCTTGCCGCCCTTGGTTTTCACAAACGTGATTTTATTGGCCGAAAGCTGCGTGCGCTGAAGGT TTTCCGCTTCGCGCCAGCGCGCGCCGGTGGCAAGACACACCTTCACCACCCGCGTTAAATCGCGATT CCCGAACGCGTCACAGGCCGCCAGCAGTGTGGCGATTTGCGGCGGTGTCAGCCACGTCATCTCGCG CTCTTTCTCGCGAAACGTCCGTACCCCCTCGAGCGGGTTCGGTAACTGCCACTCCCCCAACCGACTCA GCTCGTTAAACACCGCACTGAGATACTGCTGCTCACGGTTCACCGTCACCGGCTTCACCTTCCAGTTC GCCGGATCGCGCGTGTAACCATTATCGATTTCGCCACTCAAACGCCGATCGCGATAGTGCGCCCACA CTTTCGCGGTCAGCTCCGTCGCCAGTGGGTCGCCGAGCCCCCGGCAGACAATCTCCAGCTTAGCCAT CCGCGATTTCCCGGCCGCCAGCGACTGCCCATGCAACTTAAACCACAGCGCAATCAGATCGCTCAAC CGACGGCGGTCCTCTTnTCGCCAAGCCAGGGCTrCTCCTGCGCCTCTTGCTTCGTCCAGCGCTCAAA TGCCTCGGCTTCACCTTTGGTAGGAAACTGCCGACGAATACGTCGCCCCTCCCGTCCCTGCGGATAG AGCTCGCACAGCCACTTCCCATTTTTCTGTTTGCTGACCGTCATGGTTACCCTAAACGTGTTTTTCCAG CGTAAAAATCACCACGCCAAACGGCTCAATCTCTGTCACGTTGCACTCAAAATCCGCCGAATGATTGG TAAGACGCACCTTGCCGCCCGGCAAGCGCACCACATCAAACACATCCAGCACATCATCAATGCGGAT AAACCAGCGACCGTTACCGATTTTTTGCGCGGAAGTGTCCACCAGCCAGCCCGCATTCAGCCCTTCAA TAAAATTCAGGCTGTCGACGGACGACGGTGCCAGGCTACGATCCAGCAACCACATGCCCGCCTCTTT CAGTTCGCCGGACTCATGGCGATATTTCGGGATACTCAGCGCCGCCTCGTGTTGCACAGCCGACGTG GACGCGGCGGGATACATCTCGCCTTTGCCGGTCGCCAGCCAGTTGAGCGACACGCCGGTATCCAGC GCGCACGTCACCACCACATCGCCGGGGAAAAACTCACGCCGCACCCAGGTGCTGATGGTTCCGGAA GAGATATCCAGCAAATCACCGAGATCTTTTTGCATCTGAAACCCGTATGCATCGAGGATTCGGCGCA GCACCGGCCGCCCGCCGGTCGATAAAATCTCGTTATACAACGCTTTGCCTTTCGGTGCGGGTTTATCG GTTAATTTTGCATTTGCAAACTCACCTTCCACCAGCCAGTGCAGATCGGCCCCGGTATCGAGCGCACA TTTGACCACTGCTTTATAGGGCACGCTCTCGCGCTGGATCCAGCTACTGATATTGTTCGCCGGCACAT CCAGCGCTTCGGCGAGCGCTTTCTGGCTGGAGACGGAATAGGACGCGGCAATGCGCTCAATCACAT Attorney Docket No. PIV-00053 WO

[0313] CCTGAACACTCATCTTGTTTTCACTCATGCATACCAACTTCGAAAAAGTGATTTACACAATCGCATTTG CGATTTATATTGCTGTGCATAGACCAGAATGCACACCAGTGCAGTACATTTCAAACCACAGGAGATA ATGCGATATGTCAGATGCAAAATCAACGCCGTCTCATCCGTTATCCGACTCTCAAATGCAATCCAGAA ATTTCACCCATGGCCAGATGGATGAGCTGATGTCCGCCCTGCTGCCAGCGTTGCAAACGCTGATCCG CTCGGCGATGTCCGACGCGATGACGGTGAAAGATTTCGCCGCCATGCGCGGTATCAGCGAACGTCT GGTCTGGCAATGGCTTGACGAAGGCATATTGCTCAAAGCCCCGACACGCGACTATTCCGTGGCGCAA CATGCCGGGAAACGCAGCCGTACGTTGATCAATGTGAAAGCCTGGCGCGACAAATTAACGCAGCAG GCCATCGACTGCCGCTATATCGATCACCGCGCCGGCACATCCTTTAACTGAGTTTGCTTTTGCAAGCT CAGAAGAAAACAGGAGAGAGTATGGCTATCACAGCAGACCGCATCAGCGTACCGCTGAGCGCCGG AGAACGGCTTAACGGGCTGAACCATATCGCCGAACTCCGCACCAGTCACTGGGGCGATAGCTGGCC CGAAGTGGCGCGTTTTATCGAGGATTTGTGCGATAAGCGCGACGCGCAGTACGAGGAAAACAGCCG TGCACTGGCGGCGATCTTCTTTCTCGCCCGCGTGCCCGCGGCCCGCCAGGCTCTCGGCCCACACGCA CTGACGCTTGAAGAGAAGAGAGCCGTGATCAGTGCAATGAACCATTTCCGCGCGGTCGTCAGCCTGT TCCCGAAACGCCTGACCCTGCCGCGTTAACCCCCAATCCGTAAGCCATGGCGTCCACCCGCCAGGCC CCCTTTTGCCCGCAATCAGGAGAAACCCAATGCACCACACCCTAAGGAGCGTTACCGATGGCCGATG AAATGGATCTCGTACAACAGCGTGAGCTGGAAGAGCGCGAACGGTACATCCGCCAGGCACGCAACC GCCTGCTGTTACCGTCGCGTCTGACCTGCGAAAACTGCGATGCGCCTATCCCCGAAGCCCGCCGTAT GGCGCTGCCCGGCGTGACCTGCTGCGTAACCTGCCAGCACCGCGCCGAACGGCAAGATAAACACTA CAGGAGGCGCTAATTGGCGATCTCGTGGGCTTTTCCGTGGAACGCGCCGCGTCCGGCGATTGCCAG CCCCTACCTCACGCATGCCGAACTCCACCGTCGCAATCAGCTGATTGCGGCGTTGTTGCAAGCCCGTC ATGCGCTGGCGCTGCAACCGGAGTGTGTACGCCTGCCAATCATGCGCACGGCGGATACGCTGGAGA AGCACCACGGCGAGGCGCGCGCCAACGCCTGGCTGCGCCACGTTGTGGCGCGTACGCTGCCCCGCC TCAACGCGGTCACACAGCGTTACCAACTCACCTGCATTCACCAGCGCGTATCGAAAGCGGTATTTAAT GGCCACTTCGATACACCGGTTCAGCGCGACCTCGCGAGCCGGCTGGTGAATTTACTCGCCCGCTACA ACCGCCTACCGGACATGACGAAAAAAGCGATCGACCGGCTGTCGGCGGACATCGCCAATTTTATTCG CGGCGAACTGGCAGATATCGATAACGCCGACGACGCCGAGCTGAAAACCCTGCATCGTTGGTATAT GCGCGCGGGGATCATCGCCCTGCAATTCAACGTCACACCGCCGCACTGGCAACGAGTCACCCGCAAA CTGGCCAACGCCAGCGACCTCGCCCCGGCCGTGATCCGCCTGTTCAGCGAACAGTGGTGGCGCGGG CATTTACGCCGCATAGCTGCACAGTGGCGGGAACATCTCCAGATCGCACTGGGCAACGTCAGCAAG AAAAAACACCCCTACGCCAGCCAGGATTGCGTCAGTGCCTGGCGCGAGCAAAAACGCCGTAACCGT GAGTTTCTCAAAGGCATGGAACTCGAGGACGAAGAAGGTAACCGCATCAGTCTGATCGATAAACAC GATGGTTCGGTAGCCAACCCGGCGATCCGCCGCAGCGAGCTGATGACGCGCATTCGCGGGTTCGAG ACCATCTGCCAGAACCTGGGTTACGTGGGCGAGTTTTACACCCTGACCGCCCCTTCCGCCTGGCACG CCACCACCCGCACCGGCTACCGCAATCGGAAGTGGAACGGTGCCAGTCCGGCGCAGACCCAACGTT ACTTCACCGCGCTGTGGGCGCGTATTCGCGCCAAGCTGCACCGCCATAATCTGCGCGTGTTCGGCAT TCGCGTGGCGGAGCCGCATCACGATGGCACCCCGCACTGGCATCTGTTGCTGTTTATGCAGCCTCAG GATGTTGCTCGCGTACGCGATATTACGAGCGAGTACGCAAAACTTGAAGACAATCATGAGCTTAATA GTGAAAAAGCGAGGAAGGCGCGCTTTGACGCGCAGGCGATAGATTCACAGAAGGGCAGCGCAACC GGTTATGTGGCGAAGTACATCGCTAAGAATATCGACGGTTATGCGCTGGAAGGTGAAACCGATCAC GAGAGCGGTTCACCGTTGCAGGAAACCGCTTGCGCAGTGGCGGCATGGGCCTCGCGCTGGCGTATC CGCCAGTTCCAGTTTATCGGCGGCGCACCGGTCACGGTGTACCGCGAACTGCGGCGCATGGCCGAC AGCGAAACCGCACGCGGGTTAAGCGTGGAGTTCGCGCAGGTGCATGACGCCGCCGACGCCGGTGA CTGGGCTGGCTATGTGAATGCGCAGGGCGGGCCGTTTGTACGCCGCGACGACCTGCAAGTGCGCAC ATGGTACGAGGAGGAGAACATGTTTAATCAGTATGGCGAAACGATTGTAGCCATTCGCGGCCTGTA CGACACCACCGTCGGCGCGGGTTCGCCGGTGCTCACGCGGCTGAAGAAATGGAAGATTGTGCCGAA GCGTACGGATCTTGACCTTCAGGGCGCGTCTGCGCCCTCTTGGAGTTCTGTCAATAACTGTACGCCCC GGCGGGAATGGCAGGCGGGCTGTTGCCAGTCTCCTGACAAAATTGCGGCGCAATTCACAAAAAACA GAGGAGAGAAACCGTTATCAGAAAACAACAATACTTTTTCTAATCATCAACATAGGTATCAAGATGA AATATTTTTCGCTTCCCATTACTATTAGGCTTATGATACTGTACATCCATACAGTAAAAGCACATGAGG GGATTCATGGTGTCTGAACGTTTCGAAAAATCGCAGTTTAATTGGGCGTGCGTCAAACTGATTACTG Attorney Docket No. PIV-00053 WO

[0314] ATCTGGCGCTACTTCATCGCTGTACCAGGGAAGAGATGGAACTCGCCATGACCATGGTGTCGAACAT CGCACACAGCGAAACCCCAGGGGTAGCAGATAAGGAGCTTTTTTACCCCGCCGAATAACGCTCTGCC GCCTTCCTCAGGCAGAGGCATTTACAGCATTGCGCGCCAGCCGCCACCGGTCTGGCGCGCGCTTTTG TTGTTCTCCCCTTTCTGCATCATTACGGCTTTCTCACGTTGTGCCAACGCTTAACCAGCCCTGACAGAT GGCGTCTGTCTGCCCCACCCACGACACTTGAAGCTGGAAACACTCCCTCCAGACCGCAAGAGATCCG AACACGATGAATGCAATTACACAACAGGGCGACACCCTCGACATCCTTTGTCTGCGCTATTACGGCC GCACGGAAGGCGTGGTCGAAACCGTACTTCTGGCCAACCCTGGACTGGCGGAACTCGGGGTGATCC TGCCCCATGGCACCCACGTTGAACTGCCGGAAGTGGCGGTCGCCACGGTGAACGAAACGGTGAACC TATGGGCATGACCATCGAAAAAGTCACCTCATCGCTGGCGTATTGGATCAGCGTCGCGCTGACGTTT TTTGGCGCGATGACGCCACAAGACTTCGCCGCCTGGTTTGGCGCGCTCGGCGTTGCCGTTACCGTGG CGGTGAACTGGTATTACCGCCGAAAGAGCTATCTGTTTCTCAAATCCTGCGCCACGAGCCAGGAGGT GATCAATGGGCTTACCCGTTAAACGCTGTAGCGCAGCGGTCGTGCTGGCGCTGGCGGCACTGATAC CGGACTTCCGCTTACTCCATACCTCGCAGCAAGGGCTGGCGCTGAITGCCGATCTCGAAGGTTGTCG GCTGCGCCCTTATCAGTGCAGCGCTGGCGTCTGGACGTCCGGTATTGGCCATACCGCCGGCGTCACG CCGAAACGCGATATCACGGAAAAAGAGGCGGCGGTCAACCTGGTCGCCGATGTCCTGCACGTTGAG CGTCGACTGGCGGAGTGCGTGACGGTGGCGATGCCGCCGCAGGTCTACGACGCGGTGATCAGCTTT GCGTTTAACGTCGGTACCGGTGCCGCCTGCCGCTCAACGCTGGCGTTTTTCCTCAATAAAAAACAGT GGCAACAGGCCTGCGATCAGCTGCCACGCTGGGTGTTTATCGACGGTGTGCGCAATACCGGGCTGG AAAATCGCCGCCAGCGTGAACGCGCCTTATGTCTCCAGGGGACGTTATGAAAACCTGGCTAATTGCC CTGCTGCTGGGCGTCTGCGGCCTGCTGTGGCTGTTGCAGCAAAACCACACGCTGCGCACGTCGCTGG CGCAGTCCCGCCAGCTAACCCAGGCGCAAAACAGCGCTATCGCACAACTGAACGCGCAGCTTACCGC CTCACGCGAACTGGCGGACAGCAACGAGCAGGCGCAGGTGGCGCTGCGCCAGCAACTGGATTCCGC CAGTGCGCAGGCGGTGCAGCGCGAACAGGCTATTACGAGGTTAATCAATGAAAACGATGCGTTTCG CCGCTGGTACAGCGCTGAGCTGCCTGATGCTGTGCGCCGGGTGCACCAGCGCCCCGCCTGCGCCTCT GCCGGTCACTGTTTACAACCGTTGCCCGCAGGTCAGTCTGTGCCCGATGCCAGCAAGTAATCCCCAA ACCAACGGCGACTTAAGCGCCGATATTCGACGGCTTGAGCGCGCGCTGGAACAGTGCGCGTTACAG GTCGAAACCGTTAAACACTGCCAGGATGAGATAAATGCTAAAACCCGATACGCTGCGCAAAGCCCTG ATGGACGCCGTCCCGGCCCTGCGCACTAACCCTGCCATGCTGCGACTGTGGGTGGATAAAGGCAGA AATATTGCCACCCTCTCCCACTCACTGTCGTTTGAAAAACAGTTCAGTCTGAATGTCGCCGTCAGTGG TTTTTCCGGCGATATCGACACGATCTTCGTGCCGGTGATGCTGTGGCTACGCGACAACCAGCCCGAT ATCCTCACCGTGGAAGCCGGCCAGCAAGCCTGTTTTAGCTGGACGCTGGTTACCAATAGCGACAGTA CGCAGGATCTGGCCCTCACGCTGCAACTGACGGAACGCACACAGGTGAAGGAAGTGGATGGCGCG CTGTATGCCGAAACGCTGCCGGAGCCGTTGCCGCCGGCGTTGGTGACCCGCCCGAAAGAGCTGTAT GTTCACGGCGAGCTGCTCAGCAGCTGGTCCTCCTGATAACCCATTGCTCCCTAGCGTTGTGCCAAAAC CCGGACAGCGTTGTCCGTTTTCCAAAGCCGGGGGCGCATAGCATCATTTCGCGTATGAATACACAAC TCTCGCTCCAGGAAATGGCCCGCGCGCTGCGCAATATGATCCGCACCGGGATTATCGTCGAAACCGA CCTTAACACCGGGCGTTGTCGTGTGCAGACAGGCGGCCTTGTTACTCAGTGGCTGCAGTGGTTAACC CACCGCGCCGGTCAATCACGTACGTGGTGGGCGCCTTCCGTCGGCGAGCAGGTGTTAGTGCTGGCG GTCGGTGGCGAACTCGACACCGCCTTTGTTCTGCCCGGGATCTATTCCGACGATCGCCCGGCACCGT CCACTTCGGCGGATGCCTGGCATGTTGCCTTCCCGGATGGCGCGGTAGTTGAGTACGAACCCCAAAC CAGCGCCCTGAAAGTCAGCGGCATCAAAACCGCCGAGATCAGCGCCTCACAATCCATTACCGCCAGC GTGCCGGAGGTGTTAGTGAAGGCATCCACCCGCGTCACGCTGGATACCCCGGAAGTGGTCTGCACC AATAAGCTCATCACCGGCACGCTGGAGGTAAAAAGCGGCGGCACGATGCGCGGCAATATCGAGCAC AGCGGCGGTTCGCTGTCGTCCAACGGCAAAGTGCTGCATACCCATCAACACCCTGGCGACAGCGGC GGCACAACAGGAGCACCTCTATGACAGTTCGTTACAGCGGATTAAACCGCACCACTGGCGCGAGCCT GACCGACACTGCGCATATCCGCCAGAGCATCAGCGACATCCTGCGTACGCCGATTGGCTCGCGGGTG ATGCGCCGCGATrACGGCTCGCAATTGTTTGAGATGATCGACCAGCCGCAAACCCCGGCGCTGGCAC TGCAAATTCAGGCGGCGTGCTATATGGCGCTGCTGAAATGGGAGCCGCGCATCCGCGTCAGCGCGG TGACCGTTGAGCGCCAGTTCGACGGCAAAATGATTGTCAACCTGACCGGCCAGCTTGCCAGCACCGG TGAATCCCTTTCGTTAACCCTTCCTGTGAGTTAAAACCATGCCGATTATCGATCTGAGCCAGCTTCCTG Attorney Docket No. PIV-00053 WO

[0315] CGCCGGACGTGGTCGAGGAGCTGAATTACGAAACTATCCTCGATGCGCGTAAAGCAACGCTGATTTC CCTGTTTCCAACGGATGAGCAAGAGGCAATCGCCCGTACGCTGGCGCTGGAATCTGAGCCGTTGACC AAGTTTCTTGAAGAGAACGCCTACCGCGAAGTGATGTGGCGCCAGCGCGTCAACGAAGCCGCACGC GCGGTGATGCTGGCCTACGCAGCCGGGAATGACCTCGATGTGGTTGCCGCCAACAGCAACACCGCG CGTCTGACCATCACCCCGGCGGATGACAGCACCCTTCCGCCGACCGCCGCCGTGATGGAGTCCGATA AGGATCTGCGTCTGCGCGCGCAGCAGGCGTTTGAAGGATTAAGCGTCGCCGGGCCGGTGGGCGCG TATGAATACCACGGGCGCAGCGCCGACGGACGGGTGGCGGATATCTCCGTGGTCAGTCCGAATCCG GCGTATATCACCGTGTCGGTGCTCTCCCGCGAAGGCGATGGCAGCGCCAGCGACGAACTGGTCGCC ATCGTCGATAAAGCCCTAAACGCGGAAGATGTGCGCCCGGTGGGTGATCGCGTGACGGTGCAAAGC GCACAGATTGTACCGTACCAGATTAACGCCACGCTCTATTTCTACCCGGGACCGGAATCCGAACCGA TTCGCCAGGCCGCCGAAGCGCAACTGAAAGCCTATATCAATGCCCAGCGCCGACTTGGCCGCGACAT TCGCCAGTCGGCGATTTATGCCGCCCTGCATGTTGAAGGCGTGCAGCGTGTCGAGCTGAGCGCCCCG CAAAGCGATCTGGTGCTCGACAAACATCAGGCGTCGTATTGCACCGCGTGGAGCATTCAGGCCGGA GGTACCGATGAATGATGACCGCCTGTTACCGGTTGGCTCCTCAGGGCTTGAGGTAGCGGCAACCAC AGCGGCGGCGCAAATCGAACGTGTGCCGGTGCCGCTGCGTACCTTGTGGGACCCGCAAAAATGCCC GGCGGAGCTGCTGCCTTACCTGGCGTGGGCGCTGTCGGTCGATCGCTGGGATTACACCTGGCCAGA AGCCACCAAACGCAACGTGATTGCCGCCTCTTTTTATGTTCATCAGCACAAGGGCACACTCAGCGCCC TGCACCGTGTTGTCGAACCGCTCGGTTTTCTGATTGAGCTGCGCGAGTGGTGGCAGGACAACGCCGA GCCTGGCACCTTTAAGCTGGTGATTGGCGTGCAGGATCACGGCATCACCGAGGCGATGTACGAGGA GCTGGAGCGGTTGATTGATGACGCCAAACCCGCGAGCCGCCATCTCACCAGCCTGAATATTAGCCTG AGTACGCAGGGTGAGTTTTATGTCGGTGCGGGCTGTTACCTGGGTGAAGAGCTGACGGTTTACCCCT ACGCGCCGGAAGAGATTGTTGTTGGCGGTGAGAGCTTCCCGGCTTCCGCTGTTCATTTGATTGATGA CCTGAGAGTTAACCCATGACCACCAAATATTTTGCCATTCTGACCAACCTGGGGGCGGCGAAACTGG CGAACGCTACCGCGCTTGGCCGTCAGCTCAAACTGACGCAAATGGCGGTCGGCGATGCTAACGGCG CATTACCCACGCCGGATGCCGCGCAAACCGCATTGATTAACCAGAAACGTATTGCGCCACTGAATAC GCTGACTCTCGATCCCGGTAATGCCAGTCAGATTATTGCCGAACAGGTGATTCCGGAAACCGAGGGC GGTTTCTGGATCCGCGAAATCGGTCTGTATGATGATGAGGGCTCGCTGGTTGCCGTCGCTAACTGCC CGGAGACCTATAAACCGCAGATGCAGGAAGGCAGCGGTCGCACGCAAATTGTGCGCATGCTATTGG CGGTATCGTCGCCGTCGTCGGTGACGCTGACCATTGACCCGGCGGTGGTGCTGGCCACGCGTAACTA TGTGGATGATAAAGTCATTGAGGTGAAAGCCTATGCAGACAGCCTGCTGGCGAATCACCTGGCTGCT GCCAACCCGCATCCGCAATACGTGAAAGTAGCGGATATCGCCTACTACACGCCGATTGGCGTGCCGG TGCCTTACCCTGCGGCAATGCCGCCTGACGGCTGGCTGTTATGCAATGGCGCGGCATTTGATAAAGC CACCTGGCCTGGGCTGGCAGCGGTATACCCGTCTGGCGTGTTGCCTGATCTGCGCGGCGAGTTTATT CGCGGCTGGGATAATGGGCGCGGAGTGGATGCGGGGCGTGTGCTTGGTCAGTGGCAGGGTGATGC GTTGCAGCAGCATACGCATGATTACTACACCATTTATCCGGGAGGAGATACTGACAGGGGATCGGC ATCCTCCATCTTCTCCATTGATAACGCCACCTGGTCAACCACCAATGGTGCAGGCGGCGGGGCCCGC GTTGCAGCAGAAACGCGCCCGCGCAACATTGCCTTTAACTACATTGTCCGGGCGGCATAGCGCCCGT ATTGCCGTTCTGTTAAACACAGCGCCATCTGGCGCTnTnTTATCCCTCGTCGTTGTGCCAGCCTGTA ATAAACCCCGTTAAATGGTGTTCGTCTGTGGCTGAACGGAAAATAACGCCTGTCTTAACCGCGATGTT TCATTCATCCACAAACAACCAGAGAGTTAACGCATGTCCGCAAAATATTTTGCCATTCTGACCAGTCA GGGTGCAGCGAAGCTGGCGAACGCAACGGCGCTAGGCACAAAACTTACTCTTACGCAGATGGCGAT CGGCGATGCCAACGGCGTGTTGCCGACCCCCGATGCTTCGCAAACGAAACTGATTAACCAGAAACGT ATTGCGCCGCTGAACCGGCTTTCTGTTGACCCCAGCAATGCCAACCAGATTATTGCCGAGCAGGTGA TTCCTGAAACCGAAGGGGGTTACTGGATCCGCGAAATCGGCCTGTATGATGACGACGGCGTGCTGA TTGCCATTGCCAACTGCCCGGAAACCTATAAGCCGCAGATGCAGGAAGGCAGCGGGCGTACGCAGA CGATTCGCATGGTGGTGGTGGTCTCGTCAACGGCGGCGGTCACCCTCAAAATTGACCCGTCGGTGGT GCTGGCCACACGCCAGTACGCTGACGACCTGATGAATAATCATATTAAGGCCAGTAACCCGCACACT CAGTATGCGCCGCTTGCCAGCCCGGTATTCACCGGCGCGCCGACCGTGCCCAATACTCCTGTCAGCA ACTACGGGCAGCAAATCGCCAACACCAAATATGTGCGTGATGTTATCGCCGATTATGACGGCATTCT TCCTGTCGGCGCGCCAGTCGCCTGGCCGCAGGCAACACCGCCGGGCGGTTGGTTCAAGTGCAACGG Attorney Docket No. PIV-00053 WO

[0316] TGCCACGTTCGATAAAGTGCAATTCCCAAGGCTGGCTGCAGCCTACCCTTCTGGCGTACTGCCGGAT CTACGTGGGGAATTTCTTCGTGGCTGGGATGATGGACGTGGGGTAGATGCAGGAAGAGCGTTATTA ACTGCACAAGGAGACGCTATCCGCAACCTGGTGGGGTCGATTCCTTGTGTTGCCCCCTACGGATTCG AGCAACATGTCAGCGGGATTTTATACGGAATGGGAATGCCACAAGGCGGGGGATCGTCAGAAACCC TGGCTAACGGATATGGAATTGGTGTTAACGCCTCACTCTCCGTCCCTACCGCGGCAGAAAACCGACC GCGCAACATTGCATTTAATTACATCGTGAGGGCTCAATAATGTCGCAAGCAACGCTGAACCAGAACC TGATTGCCACCGCTGCCGGGGAGATCACTGTCTATAACTTTGATGGTGAGACACGCGAATACTACTC TACGTCGGTAGAGTACCTGGCGATTGGCGTGGGTATTCCCGGCAATTCGACCGTTGAACCACCGCTG GCGGCGAAAAACGGTTTCGCGGTGTGCTGCGCGACCGATGGCCGCAGTTGGCAATACGTTGCCGAT CATCGCGGCGAAACCGTTTATAATCTCGAAAGCGGTGAAAAAACGTCTATTACCACGCTGGGTGATT ACCCGGCGAACGTGACCACGCTTGCCCCGGCAACGCCGTACGATAAGTGGAACGGCAGCACCTGGG TCACCGACAGCGCCGCACAGCATGCCGCGCAAATAGCCAGCGCAGAGCAGAAAAAAACGCAGTTGC TGAATGAAGCGAAAAACACCATCAGCCTGTGGCAGACGGCCCTGCAACTGGGCCGCATTAGCGATG CTAATAAAGCGCGGCTGATCGCCTGGATGGATTATATTGATGCCGTGCAGGCAATGGATAGCGCGC AGGCACCGGATATCAGCTGGCCTGTGCAACCGGCATAATACGCAACGGGCTTCGGCCCGTTTTTTAT GCCGGGTTGTACCAGACCGCAGCCAACCCCGACCGATAGCCCCCTCCCACATCTGCACCGGACAATG TCGTTTACTTATCCGATAACCGAGAGTGAACGCATGACTGCGAAATATTATGCCATTCTGACCAATCA GGGAGCAGCGAAGCTGGCGGCCGCCACGGCGCAAGGCACCCGGCTTAACATTACCCGCATGGCAGT TGGCGACGGTAATGGCACGCTGCCCTCACCGAACGCCACGCAAACCAGCCTGGTGAACCAGAAACG CATCGGTGCACTGAATGTGCTGATGGTGGATAGCATCAACAGTAACCAGATTATCGCCGAGCAAATT ATTCCGGAAAATGAAGGCGGCTTCTGGATCCGCGAAATGGGTCTCTACGACGATGCTGGGGTACTG GTGGCCGTGGCGAACTGCCCGGAAACCTATAAACCGCAACTGGATGAGGGCAGTGGCCGCACGCAA ACCATCCGCATGGTGCTGACGATTTCGAATACGTCGGCCATCACCTTAAAAGTGGATGCCACGGTGG CGCTCGCCAGTCGTCAGTACGTGGACGACCACCTTAGCGATCATATTAAAGCCTCCAACCCACACAG CCAGTATTTGCAAATTTCCCAGTCACTGAAAGAGATAGCCGATTCGGGTCCCGCAGCCGTCGCTGCG GCGGTTAAAAACCTGGGTTTAAAAGAAGCGGCTAATGGCGTGAGTGTAGTGGGACAATCCCGCAAT GCAAGAATGTACATCAACGCAGTGTCAGCATCAGCAATCTTCACTGCAGATGAACTTGTTGTGGCAA CAGTATTGGGAGGAAAAAATTATCGCCTCGCAAACATCAAAAAAACGATTGATCTGACCAGCAATGG TGCAGGTGGAATGGATGCTGGAGCGGCACCAGTTAGTGGTTTTGTTGCTTTGTATGCAATATACAAC CCAACCACAGACACTTCGGCGCTACTTTCTGTAAACGCAACATCTGTTGTTGCGCCCGAAGTCTATGG TGGTACAAACATGCCTGCCGGGTACACTGCATCCGCCCTCGTTGCCGTTCTACCTGTGGTTTCGTCTC AGTITTCAATATGTGGTGTACAGGGCCGTTCCGTGGTCATACCGCCAACAACAACATTAAGTGGTTCA GTAGCGCCTAACACAGCTATTTCGCAAACGATGGCTGCCGTTCCATTGAATACGGTTGCTGTAGATTT ATCGGTTTCAATCGGGCTGGTTAATGTGGATACGACGGGGGTmTGTTATCGGAAGTTCAAGCAAC TTACTCGGTTCTCGTCGGTTTTCTGTTGGTTCATCATTAACAGGGGGGACAGTTAGCCAGAGCAGTAA TACAACTGTGCCCGTAATCGCGAACACACGTACATTTTATTGGCAGGTTTCAGCCGCAAGCCTCACAT ATAACGTTGGGATCCAAGGATATCAAATATGACTAATAACAGTACCTTCTATGCGAAATTAGGAGAA AACGAGAAGATTCACTACATATCTCTTGGGGAACAACTTGATCCCACTATGTTTCCTGTAGATTACAG CAGCGAGTTATATCGTGAGTTTTACTCGCGAATGCCTGCCTGGGCCCAGAGGGACATCCCTGCTCCA TCTGAAAAAGAAGCCGCCGCATAACGCGGCTTTTTTTCATCAGAGAGGAACAGGAAGCTGAATTACC GATGTCTACATCTGTTGTTTCAGCCCAGCCCCAACGCCGATAAATAGCGCCCGACCCCGCCGCCCTGG AGAATAGCACTCACCCCAAAACCACGGAGTTAAACGGATGAGTGACTATCATCATGGCGTCCAGGTC GTCGAAATCAACGACGGCACACGCGTCATTTCCACTGTCTCAACGGCCATTGTCGGCATGGTATGTA CCGCCAGCGATGCAGACGCTGAGATGTTCCCCCTTAATGAACCAGTGCTGATCACCAGCGTGCAGAG CGCCATTGCGAAAGCGGGCGTCAAAGGTACGCTCTCCTCTTCTCTGCAAGCCATTGCTGACCAGGCG AAACCGGTCATCGTTGTGGTGCGCGTTGCGGAAGGCACGGGCGACACCGCGCAGGCGCAGACCATT TCCAATATCATCGGCACCACCGATGCAAACGGTAAATACACTGGCCTGAAAGCACTGCTGACCGCTG AAGCGGTAACCGGCGTGAAACCGCGTATTCTTGGCGTTCCGGGTTATGACACCCAGGAAGTCGCCAC CGCACTGGCCTCTGTCTGCCAGAAACTGCGCGCCTTCGGCTATGTCAGTGCCTGGGGCTGCAAAACC CTTTCCGATGCAATGAAATATCGCGAAAACTTCAGCCAGCGTGAGTTGATGGTGATCTGGCCAGATT Attorney Docket No. PIV-00053 WO

[0317] TCCTCGCCTGGGATACCGTCGCGAACGCGACTTCCACCGCCTACGCTACTGCCCGCGCGCTGGGTCT GCGTGCTTACATCGACCAGTCCGTTGGCTGGCACAAAACCCTGTCCAACGTTGGCGTTAACGGTGTG ACCGGCATCAGTACTCCGGTGTTCTGGGATTTGCAGGAATCCGGTACCGACGCGGATCTGCTCAACG CGGCTGGCGTGACCACGCTTATCCGCAAAGACGGTTTCCGCTTCTGGGGCAACCGCACCTGCTCCGA TGATCCGCTGTTCCTGTTTGAAAACTACACCCGCACCGCGCAGGTTATCGCCGACACCATGGCCGATG CGCATATGTGGGCGGTGGACAAACCGATCACCGCAACGCTTATCCGCGACATCATCGAAGGCATTAA TGCGAAGTTCCGCGAGCTGAAAAGCAACGGGTACATCGTGGATGCCACCTGCTGGTTCGACGAAAC CGCCAACGATGCCGTCACCCTGAAAGCCGGGAAACTGTATATCGATTACGACTATACGCCGGTGCCG CCACTGGAAAACCTGACCTTACGCCAGCGCATCACCGATAAGTACCTGGTGAATCTGGTCTCCTCGGT CAACAGCAATTAAGGAACCTGATAAATGGCAATGCCGAGAAAACTGAAATACATGAACGTGTTCCTC AATGGCTACAGCTATCAGGGGATCGCCAAATCCATCACGCTGCCGAAACTGACCCGCAAGCTGGAA AACTACCGCGGCGCAGGCATGAGTGGTGTCGCGCCTGTCGATATGGGGCTGGATGACGACGCCATG TCGATGGAGTGGACGCTGGGCGGTTTCCCGGATACGGCTATCTGGGAACTGTATGGCGCAACCAGC GCAGACGCCGTCCCTGTGCGTTTCGCCGGTTCCTACCAGCGCGACGACACCGGTGAAACGGTGGCA GTCGAAGTGGTCATGCGTGGTCGCCAGAAAGAGATCGACACAGGCGAAAACAAAGTGGGCGAAGA CACCGAATCCAAAATCTCCGTCGTTTGCACTTATTTCAAACTGACGATGGATGGCAAGGAACTGGTG GAAATCGACACCGTTAACATGATCGAAAAAGTCAACGGCGTAGACCGCCTCGAGCAGCACCGCCGC AATATCGGCCTGTAATGCGCATCCCGGTCAGCCTCGCTGGCCGGTTCTTCCCCCGCTAATGGAACCAC GAGGAACTCATGAGCAACGAAACTGACAACGTCATCACCCTGGAAAACCCGATCACACGCGGCGAA CAGAGCATTACTGTTATTACCCTGATGAAACCGAACGCCGGCACGCTGCGTGGCGTGAGCCTGGCG GCAGTGGCGAATGCCGAAGTGGATGCGCTGATCAAAGTGTTGCCGCGCATTACCTCGCCGTCGCTTA CCGAGCAGGAAGTGAGCGCGCTGGACCTGGTGGATATGGTGGCGCTGGCCGGCAAGGTGGTTGGT TTTTTGTCACCGGCTTCGGCACGGTAAGCTTTCCGGCGAACCTGTCGGTTGACGATCTGATGGCGGA TATCGCGGTGATTTTTCACTGGCCGCCGTCAGAGCTCTATCCCCTGAGCCTGAGCGAACTCATCACAT GGCGCGAAAAAGCGCTGCAGCGAAGCGGAAACACACATGAGTAACAGCGTAAAAATAGAAGACTT GCTCACGGCTGTTGATCAGGCGACGCGCCCGTTTAAAACCTTACAGACCGCGAATGTGTCGCTGTCT GATCAGGTTAAAGAGACGCAGACCACCTTACGCGGCCTGTACAGTCAGGTGGCGCAAATCGACGGT TTTACCCGAACGCAAAGCGCCCTGACTGCCGTCAGCCAGGCGCTGAACGTCGCCAAAGAGAACACCC GCGCGCTGGCATCTGAACTGAAAAAAGTGGACGAGCCCTCCCGTTCGCAAATAAGCGAACTGGAGA AAGCCCGCAGCCGCGTCAGCACCCTGCAACAACAGCATGATGACCTGCGTCAGTCGGTGAAAAACC AGCGCCAGGTACTGAGCCAGGCAGGCATCAGCACCCGCGCACCGGCAAGCGCACGGCAGCGTCTGC AAAACAGCATCAACGAGCATACCGGGCAACTGACCACCCAGCGCGATGCGCTGGCGCAGGAGAACC GGCAAAACCGCATGGCGAAAATCAAAGCGGGACAGCAATCCATTCTGGGTATTGCCGGGAACCTCT CCACCGTCGGCAAAACCGGCATGTCGGTTGCTACCACCGGGGTTAACCTCGGCAAAAAACTGCTGCA ACCGGGTTACGAAGCGTCGCTGAAAGAGGCACCGGCAGCCACCCCTGCCAGTCCAGCCAACCCCGG CGCGAACGTGGCAACCGCGCAGAGCGGCGGCAATCTCGGCACCGATTTACAAGCGCTGCAAGCGGC GTATCAGTCGCTGAGCGTGGATATTTTCCGCACTCAGGAATCCTCTTTACGCCAGCTGGTACAAACCG CTACTCAGTATCTGGGCCAGCTACAACAGTGGGTACAGAACAACCAGGGACTGGTGCAAACCTTCG GCATGATCGCCACCGTGGTGGTTGGCGTCACCGGGGCCATTGGCACCGTGGCCAGCGTTATTGCGC CCGTCTTTACGGGCATCAGTACGCTTATCACCATCGCCACCACCCTTGGCAGCGTCTTTACCACCGTG TGTGGCGCTATTGTAGCGGTACTTGGCTCGCTTACCCTGCCCATCATCGGCATTATCGCCGCCATTGC CGGCATAGCGCTGGGCATTTACGCCCTCTGGGAACCGATTAGCGCCTTCTTTGGCGGCGTGGTCGAA GGTATCAAAGCCGGATTAGCCCCACTTGCCGGACTGTTCGCGCCTTTACAGCCCGTTTTCGACGCCAT TGGCAACGGCCTGACGTGGATAAAAGACCTGTTTGGCGCGCTGCTCTCCCCGATTACATTCAGTCAG GAAGCGCTAGAGAAATGCGGCAATGCCGGGCAATCCTTCGGCAAGTTTCTGGCCGATGGGCTGATC GGTTGCTTATGGCCGCTGAAATTGCTGATCGACGGCGTCGGCTGGCTGCTGGAGAAACTCGGCATC GTGAACAAGCAACCGGCGATTACGATCCCCAAACCGCAGGTGGACGCAACGGCGGTGGGGACAAC ATTCGGTTACGTCCAGCCAACCAGCACAATGCCGGGATTCAACAACTACCAGGCAGCCAGACCCGCA GGCGGTAATTCGTATGTCGACCAGAGCAAAACGGAATATAACGTCACGCTACAGGGCGATATCACC CCCGGTAGCACTAACGAGCGCTACCTGCAGGATCTGTTCCGCCAGGACGCGGCGAACAAACGCAAC Attorney Docket No. PIV-00053 WO

[0318] AGCACCCTTTCGCAATTCTATCCATCAGGAGGGTTTTAATTATGATGCTGGCGCTGGGACTTTTTGTC TTTATGCGGCAAACGCTGCCGTTTCAGAACCTGAACCGCACCTCGACGTTTACCTGGAGCGGCAACG CGCGCGTCGGTAAACGCGCGGCGTATCAATATATCGGTCCGGGCGAAGATACTCTCGAAATTACCG GCGCGCTCTACCCGGAACTGACCGGCGGCGTGCTGTCGCTGGCGGCCGTGCGTCTGATGGCTGAAC AGGGCGGACAATGGCCGCTCATCGACGGCACGGGCATGATTTACGGTGTGTATGTCATCAGCAGCG TCAAAGAGAACGGCTCCGAGTTTTATAGCGACGGCTCGCCGCGCAAAATCAGCTTCACCCTGAATTT AACCCGGGTCGACGAGTCGCTGCTCTCCGAGGCAGAAGCGCTTTACACGAAAGGCAGTAAGCTGCT CGACGATGCCACCAAAATGCTGGGGAGTCGCGGATGATTGACGCTCTCGACAACGATCTGACGCGG GGAGATGCCCCCGCGTTTAAGCTGGAGCTGAACCAAAAACCCGCCAGCAGCAGCGCCTCTGCGGCA AAAAACAACACCAGTAAAGACGAAAAAAAAGACATTACCGCCAAGCTCGCCCCGCGATTAATCGCG CTCACGGTCACGGACAACCGCGGTTTTGAAGCTGACACGCTGACACTGACGCTGGATGACAGCGAC GGCAAAATCGAGATGCCAGAGCGCAACGCGCAGGTTAACGTCTCGATTGGCCGACAGGGCTCGGTA CTGACCAATATGGGAAGTTTCATTATCGACCAGGTCACGCACCGGGGCACGCCGGATCAACTGGTGA TTACCGGGCGCAGCGTCGATTTTCGCAATGACATGAATACCCCAAAAGAGTGGTCATGGCATGACAC AACGCTTGGCGAGATTGTGAAAGATATTGCCTGGCGCAATTCGCTGAGCGCCAGCGTCTCGCCGGC GTTGGCCGGTATTACGATAGCGCATATTGACCAGTCCAAAGAAAGCGACATCGCCTTTCTCACCCGG CTGGCTATCCGCAACGGTGCTGAAATCGCCGTAAAATCGGGCGTACTGATTTTTCTTGTGCCGGGTA AATGCGTACGAGGCGGTAAGGCCATCCCAACGGTGACCATCAGCCGTAGCGATGGCGATGCTCATA CGTTTGATTTGGCCGATCGGACAGCATACGGCAGTGTCATCGCTCGCTGGCAGGATACGAAAACGCC GCAGAAACAGGTGAAACAGGTAAAACTGACGCGCAAAAGCGCCGGCACGCAGCAGGCAACAAAGG ATTACCTCGCCGGTTCACGCGAGAATGTCAGCATCCTGCAAAACATGTTCGCCTCCAAAGATGAAGC CATCCGTGCGGCAGAAGCGGAATGGCAACAACGTCAGAGAAATGCAGCGCAATTTAGCCTGACTCT CGCGCAAGGACGGGCCGACATCGCGCCGGAAACGCCTGTCCGGTTATCGGGTTTTAAGGATGTGAT CAATAAAACGCCATGGGTGATCAAAAAAGTGACTCATGGCATTGATAAAAAAGGGTTTATTACAAAA CTGGAGCTGGAAGCCGACATCCGAAACGTGGAGTACGACATGGAGGAATCTGGCGCCTGAAAACGT CAAATGCAATTGTTAACTTGCATTTGCGAGTTAATGGTTTATTATTACTGGCATAAACACGCTGGAGA AATCACAAAATGATGCATTGCCCGTTATGCCAGGAAGCGGCTCATGCGCGTTCAAGCCGCTATCTCA GTACCGAAACCAAAGAACGCTACCACCAGTGCCAGAACATCAACTGCGGCTGCACCTTCGTGACCCA CGAATCGTTGGCACGCTACATTGTCCGTCCTGGCGCGACACAAGCCGCACCGCCCCACAGTGCGCGG TAA

[0319] Example 9: Confirmation of prophage grounding

[0320] The modified strains were then tested for prophage activation by the addition of mitomycin C to cultures, followed by sequencing and read coverage analysis. Increased read coverage (as seen in the two replicates of WT 5812 strain) signified replication of a triggered prophage (FIG 6A). Replication levels were inhibited in the strains containing the prophage grounded by recombinase deletion (FIG. 6B) and 3' edge trimming to delete attR (FIG. 6C) in comparison to the wildtype strain (FIG. 6A).

[0321] Example 10: Assessing competitive fitness index of strains with grounded phage

[0322] As a control, a complete deletion or 'knock out' of the prophage P-5812-01 was made (CD KO), to which a plasmid encoding mCherryV2 (strain 5812-8679) or GFP (strain 5812-8678) was added. K, oryzendophytica strain 5812 was found to contain several phages, in addition Attorney Docket No. PIV-00053 WO

[0323] to the one described above. Three different phage recombinases were identified (three prophages were identified in 5812 each with their own recombinase). All three of these recombinase sequences were deleted from strain 5812, creating a grounded prophage (grounded CP) strain of 5812, to which either a plasmid encoding mCherryV2 (strain 5812-8686) or GFP (strain 5812-8687) was added.

[0324] Phage grounded strains (containing deletions of all three recombinases as described above) and complete knock out strains, tagged with either GFP or mCherryV2, were applied to SOB agar plates supplemented with kanamycin (50 pg / ml) and incubated at 30°C for 24 hours. After the 24-hour incubation, Falcon test tubes were each filled with 2mL SOB supplemented with kanamycin (50 pg / ml), and growth from each agar plate was scraped into a test tube. ImLfrom each test tube was transferred to a cuvette and OD590 was read against an SOB blank. Test tube cultures were then diluted to an OD590 of 0.05 with SOB. These OD590-normalized samples were combined in a pairwise manner at a ratio of 1:1 in a deep 96-well plate (600 pl of each culture with each well having a total volume of 1200 pL). These combined samples were run on the flow cytometer and plated on SOB to determine the strain ratios of timepoint zero (TO).

[0325] The 96-well plate was sealed with breathable film and placed in a 30°C incubator at 250 rotations per minute (RPM) for 24 hours. Three biological replicates of the co-culturing were performed. After the 24-hour incubation, cultures were diluted and run on the flow cytometer and plated on SOB to determine the strain ratios of timepoint one (Tl).

[0326] Flow cytometric measurements were obtained on an Invitrogen Attune NxT (ThermoFisher). The machine was calibrated to measure the number of particles in 15 pl of a 200 pl sample, using a flow rate of 25 pl / sec. Samples were subjected to 200X dilution to reach the appropriate events per sec. The yellow laser (561 nm, 50 mw) and the blue laser (488nm, 50 mw) were used to excite the fluorescently tagged strains. Counts for cells with GFP fluorophore were recorded in the BL1-H channel, while counts for mCherryV2 tagged cells were recorded in the YL2-H channel. Using the histograms from the BL1-H and YL2-H channels, gates were applied to the fluorescent intensities for each fluorophore using single strain controls, following the expected fluorescent intensity distribution determined by previous experimentation. Spectral overflow from the two fluorophores was adjusted by applying compensation. The same gates and compensation were applied to all the samples. Attorney Docket No. PIV-00053 WO

[0327] The competitive index was determined by dividing the ratio of the strains (Strain 1 / Strain 2) at the end of the experiment (Tl) by the ratio of strains (Strain 1 / Strain 2) at the beginning of the experiment (TO).

[0328] strain 1T:)

[0329] T, strain 2 CompT

[0330] retive index — - s~tz —1

[0331] raml —Tn

[0332]

[0333] strain 2T0

[0334] Ratios of mCherryV2 and GFP strains as determined by flow cytometry (FIG. 4A) and by plating for colony forming units (CFU) are shown in FIG. 4A and FIG. 4B, respectively. In FIGs. 4A-4B, if strain 1 had an increased competitive fitness compared to strain 2, the competitive index was above 1; if strain 2 had an increased competitive fitness compared to strain 1, the competitive fitness was below 2. The two different fluorophores (GFP and mCherry) did not result in different competitive indices in either background. Regardless of which strain was tagged with a GFP or mCherry fluorescent reporter, the strains with grounded prophage displayed an increased competitive fitness in comparison to the strains having a complete knock-out (KO) of the prophage sequence (FIGs. 4A and 4B), confirming that the grounding strategy described herein can result in a more competitively fit strain than a strain including complete deletion of activatable prophages.

[0335] Example 11: Evaluation of product production: nitrogen excretion in strains with grounded phage

[0336] K. oryzendophytica strain 5812 discussed above is a nitrogen-fixing bacterium. To confirm that K. oryzendophytica strain 8252 having the grounded phages through deletion of three phage recombinases (AxerD, AxerC2, AxerC3) retained the same nitrogen-fixing properties, ammonium secretion of the strains was determined.

[0337] Secretion of fixed nitrogen was measured in wildtype (WT), ammonium secretion mutants of K. oryzendophytica strain 5812 with intact phage and with grounded phage. Ammonium secretion was measured via the method provided in Bloch et al. Journal of Experimental Botany 71.15 (2020): 4591-4603 (2020). Briefly, strains were cultivated on SOB agar for 24 hours at 30°C. Single colonies were transferred to 1 ml liquid SOB, shaken at 200 RPM for 24 hours at 30°C. 50 pL were transferred to 1.15 mL of defined medium supplemented with nitrogen and incubated under the same conditions. 100 pL of this culture was then transferred into 900 pL nitrogen-free defined medium and incubated under anaerobic conditions supplemented with a gas mixture of >98.5% nitrogen, 1.2-1.5% Attorney Docket No. PIV-00053 WO

[0338] hydrogen, and <30 ppM oxygen and incubated at 1350 rpm at room temperature for 72 hours. The initial biomass was compared to the end biomass by measuring optical density (OD590) for each culture using a spectrophotometer. Cells were then separated by centrifugation and supernatant from the reactor broth was assayed for free ammonium using the Megazyme K-AMIAR Kit normalized to biomass at each time point as per the kit instructions provided by the manufacturer.

[0339] When the three identified prophage in the ammonium secretion mutant strain were grounded by deletion of the three prophage recombinases, there was no detectable difference in ammonium secretion between the ammonium secretion mutant and the ammonium secretion mutant with grounded prophage (unpaired t-test P-value > 0.05) (FIG.

[0340] 5). Accordingly, the grounding of the prophage had no deleterious effect on product production.

[0341] Example 12: Identification and grounding of a prophage in a Kosakonia arachidis Strain.

[0342] A prophage was identified in Kosakonia arachidis strain 1661-5402, a nitrogen-fixing bacterium that excretes nitrogen. K. arachidis strain 1661-5402 contains an active prophage including an unaltered attL site and predicted recombinase. (See FIG. 7A.) A culture of the strain was treated with mitomycin C (2 pg / mL) to trigger the prophage. The culture was then incubated overnight. DNA was isolated and analyzed for the presence of a prophage, revealing an increased read coverage across the region suspected of encoding the prophage (FIG. 7B). The prophage region was sequenced, and the predicted attL, attR, and recombinase sites were identified by the method described in Example 4.

[0343] To generate a grounded prophage, the attL and predicted recombinase, located adjacent to each other, were deleted from K. arachidis strain 1661-5402 to generate K. arachidis strain 1661-11364 containing a grounded prophage (FIG. 7A). The engineered K. arachidis strain 1661-11364 did not show increased read coverage over the prophage region when exposed to mitomycin C and incubated overnight (FIG. 7C).

[0344] Example 13: Determination of growth rate in Escherichia coli strains.

[0345] A non-engineered, bacterial model strain Escherichia coli (BG) and two modified strains of the parental E. coli strain (duplicate strains " I-C6", " I-C7"), were obtained commercially (E. coli Keio knockouts, Horizon Discovery, Cambridge, United Kingdom). E. coli strain BG has been reported to contain nine prophages, including a prophage designated Attorney Docket No. PIV-00053 WO

[0346] CP4-57. Strains I-C6 and I-C7 are modifed to remove the prophage recombinase region from the CP4-57 prophage. The remaining prophages are intact in these strains. Final yields of bacterial growth after an overnight incubation showed no significant difference in growth of the I-C6 and I-C7 strains compared to the BG strain after five hours of growth.

[0347] Example 14: - Identification and grounding of a prophage in other bacterial strains.

[0348] A bacterial strain of interest is identified. The sequence of the bacterial genome is determined. Using the prediction procedure described in Example 4, likely candidate prophages are identified, and the prophage recombinase sequences and attachment sites are indicated. Following the methods described in Example 2, prophage recombinase sequences and / or prophage attachment sites are deleted from the genome, but the remaining portions of the phage sequences are left intact. The resulting strains are shown in Table 7, where "+" indicates that the strain contains an active genetic element, and indicates that the indicated genetic element was deleted. A cell culture is grown to determine the effect of the deletions on the competitive fitness of the cell, as well as any production of compounds of interest, and culture stability over time. The data is compared to that of a similar cell that does not have a modification of the phage sequence(s), and also to that of a cell that has a complete deletion of the active phages. By use of this method, the modified cell has a higher level of competitive fitness, and increased culture stability, than the wild type cell.

[0349] Table 7. Strain Engineering

[0350] Strain Phage attR attL

[0351] Recombinase

[0352] A - - B - + 4- C - -F - D - - 4- E -F - - F ■F - G ■F +

[0353]

[0354] Attorney Docket No. PIV-00053 WO

[0355] OTHER EMBODIMENTS

[0356] It is to be understood that while the invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

Claims

Attorney Docket No. PIV-OOO53 WOWHAT IS CLAIMED IS:

1. A method of increasing bacterial culture stability, the method comprising:a) identifying a bacterium having a prophage;b) modifying an attL DNA sequence of the prophage, an attR DNA sequence of the prophage, and / or a nucleic acid sequence encoding a recombinase polypeptide within the prophage, thereby producing a modified prophage; andc) generating a bacterial culture using a bacterium having the modified prophage; wherein the nucleotide sequence of the modified prophage comprises from about 51% to about 99.99% of the nucleotide sequence of the prophage,wherein the modified prophage has a decreased likelihood of self-excising from the genome of the bacterium having the modified prophage compared to the likelihood of the prophage self-excising from the genome of the bacterium having the prophage, and wherein the bacterial culture generated from the bacterium having the modified prophage has increased culture stability compared to the culture stability of a bacterial culture generated from a bacterium having the prophage.

2. The method of claim 1, wherein the culture stability is measured by a decrease in prophage activation after treatment with prophage activating conditions.

3. A method of decreasing the likelihood of prophage replication, the method comprising:a) identifying a bacterium having a prophage;b) modifying an attL DNA sequence of the prophage, an attR DNA sequence of the prophage, and / or a nucleic acid sequence encoding a recombinase polypeptide within the prophage, thereby producing a modified prophage,wherein the nucleotide sequence of the modified prophage comprises from about 51% to about 99.99% of the nucleotide sequence corresponding to the prophage, andwherein the modified prophage has decreased likelihood of replication in a bacterium having the modified prophage as compared to the likelihood of replication of the prophage in the bacterium having the prophage.Attorney Docket No. PIV-OOO53 WO4. The method of claim 3, wherein the likelihood of replication is measured by whole genome sequencing after treatment with prophage activating conditions.

5. A method of preventing prophage excision in a bacterium, the method comprising modifying an attL DNA sequence of a prophage, an attR DNA sequence of a prophage, and / or a nucleic acid sequence encoding a recombinase polypeptide within a prophage, thereby producing a modified prophage,wherein the nucleotide sequence of the modified prophage comprises from about 51% to about 99.99% of the nucleotide sequence corresponding to the prophage, and wherein the modified prophage has a decreased likelihood of self-excising from the genome of a bacterium having the modified prophage compared to the likelihood of the prophage self-excising from the genome of a bacterium having the prophage.

6. The method of claim 5, further comprising:growing a bacterial culture in a condition that activates the prophage, wherein the bacterial culture comprises a plurality of bacteria having the modified prophage; and determining decreased prophage replication.

7. The method of claim 6, wherein the condition that activates the prophage comprises mitomycin C or late stationary phase growth.

8. A method of generating a modified prophage having an impaired ability to lyse a host bacterial cell, the method comprising modifying (i) an attL DNA sequence of a prophage, (ii) an attR DNA sequence of a prophage, and / or (iii) a nucleic acid sequence encoding a recombinase polypeptide of a prophage, thereby producing the modified prophage,wherein the nucleotide sequence of the modified prophage comprises from about 51% to about 99.99% of the nucleotide sequence corresponding to the prophage, and wherein the modified prophage has a decreased likelihood of self-excising from the genome of the bacterium having the modified prophage compared to the likelihood of the prophage selfexcising from the genome of the bacterium having the prophage.Attorney Docket No. PIV-OOO53 WO9. The method of any one of claims 1-8, wherein the nucleic acid sequence encoding the recombinase polypeptide, the attL DNA sequence, and the attR DNA sequence are completely deleted.

10. The method of any one of claims 1-8, wherein the nucleic acid sequence encoding the recombinase polypeptide, the attL DNA sequence, and the attR DNA sequence are partially deleted.

11. The method of any one of claims 1-8, wherein the nucleic acid sequence encoding the recombinase polypeptide is completely deleted, and wherein the attL DNA sequence and the attR DNA sequence are partially deleted.

12. The method of any one of claims 1-8, wherein the nucleic acid sequence encoding the recombinase polypeptide and the attL DNA sequence are completely deleted, and wherein the attR DNA sequence is partially deleted.

13. The method of any one of claims 1-8, wherein the nucleic acid sequence encoding the recombinase polypeptide and the attR DNA sequence are completely deleted, and wherein the attL DNA sequence is partially deleted.

14. The method of any one of claims 1-8, wherein the nucleic acid sequence encoding the recombinase polypeptide is partially deleted, and wherein the attL DNA sequence and the attR DNA sequence are completely deleted.

15. The method of any one of claims 1-8, wherein the nucleic acid sequence encoding the recombinase polypeptide and the attL DNA sequence are partially deleted, and wherein the attR DNA sequence is completely deleted.

16. The method of any one of claims 1-8, wherein the nucleic acid sequence encoding the recombinase polypeptide and the attR DNA sequence are partially deleted, and wherein the attL DNA sequence is completely deleted.Attorney Docket No. PIV-OOO53 WO17. The method of any one of claims 1-8, wherein the recombinase polypeptide encoded by the modified nucleic acid sequence, the modified attL DNA sequence and the modified attR DNA sequence are inactive.

18. The method of any one of claims 1-17, wherein the nucleic acid sequence encoding the recombinase polypeptide, if present, comprises a xerC2 gene or a portion or homolog thereof, a xerC3 gene or a portion or homolog thereof, or a xerD gene or a portion or homolog thereof.

19. The method of any one of claims 1-18, wherein the bacterium is a gram-negative bacterium.

20. The method of any one of claims 1-18, wherein the bacterium is a gram-positive bacterium.

21. The method of any one of claims 1-20, wherein the bacterium is a nitrogen-fixing bacterium.

22. The method of claim 21, wherein the nitrogen-fixing bacterium is of a genus selected from Azospirillum, Kosakonia, Klebsiella, and Paraburkholderia.

23. The method of claim 21, wherein the nitrogen-fixing bacterium is of the species Kosakonia oryzendophytica or Kosakonia arachidis.

24. The method of claim 21, wherein the nitrogen-fixing bacterium is of the species Kosakonia oryzendophytica, strain 5812 (ATCC Accession No. PTA-127355).

25. The method of claim 22, wherein the nitrogen-fixing bacterium is of the species Azospirillum palustre.Attorney Docket No. PIV-OOO53 WO26. The method of claim 21, wherein the nitrogen-fixing bacterium comprises a modification in one or more genes involved in nitrogen fixation or nitrogen assimilation.

27. The method of any one of claims 1-26, wherein the modified prophage has a DNA sequence that has at least 90% sequence identity to SEQ ID NO: 9-11.

28. The method of any one of claims 1-27, wherein the bacterium having the modified prophage has an increased competitive fitness when co-cultured with a corresponding bacterial cell that does not have the modified prophage.

29. The method of claim 28, wherein the increased competitive fitness is determined under an environmental stress condition.

30. The method of claim 29, wherein the environmental stress condition is selected from the group consisting of nutrient stress, salt stress, osmotic stress, cold stress, heat stress, excess UV radiation, the presence of antibiotics, and DNA damage.

31. The method of any one of claims 1-30, wherein the bacterium having the modified prophage has an increased cell growth rate in comparison to a corresponding bacterium that does not have the modified prophage.

32. The method of any of claims 1-31, wherein the bacterium having the modified prophage does not contain an activatible prophage.

33. The method of any one of claims 1-32, wherein all of the phage recombinase sequences that are present in the bacterial genome are inactivated or removed.

34. A genetically engineered bacterium comprising a modified prophage,wherein the modified prophage comprises at least one modification selected from a modified attL DNA sequence, a modified attR DNA sequence, a modified nucleic acid sequence encoding a recombinase polypeptide, a partially deleted attL DNA sequence, a partially deletedAttorney Docket No. PIV-OOO53 WOattR DNA sequence, a partially deleted nucleic acid sequence encoding the recombinase polypeptide, a full deletion of an attL DNA sequence, a full deletion of an attR DNA sequence, and / or a full deletion of a nucleic acid sequence encoding the recombinase polypeptide; wherein the nucleotide sequence of the modified prophage comprises from about 51% to about 99.99% of a nucleotide sequence of the prophage; andwherein the modified prophage has a decreased likelihood of self-excising from the genome of the genetically engineered bacterium comprising the modified prophage compared to the likelihood of an unmodified prophage self-excising from the genome of a bacterium having the unmodified prophage.

35. The genetically engineered bacterium of claim 34, wherein the nucleic acid sequence encoding the recombinase polypeptide, the attL DNA sequence, and the attR DNA sequence are completely deleted.

36. The genetically engineered bacterium of claim 34, wherein the nucleic acid sequence encoding the recombinase polypeptide, the attL DNA sequence, and the attR DNA sequence are partially deleted.

37. The genetically engineered bacterium of claim 34, wherein the nucleic acid sequence encoding the recombinase polypeptide is completely deleted, and the attL DNA sequence and the attR DNA sequence are partially deleted.

38. The genetically engineered bacterium of claim 34, wherein the nucleic acid sequence encoding the recombinase polypeptide and the attL DNA sequence are completely deleted, and wherein the attR DNA sequence is partially deleted.

39. The genetically engineered bacterium of claim 34, wherein the nucleic acid sequence encoding the recombinase polypeptide and the attR DNA sequence are completely deleted, and wherein the attL DNA sequence is partially deleted.Attorney Docket No. PIV-OOO53 WO40. The genetically engineered bacterium of claim 34, wherein the nucleic acid sequence encoding the recombinase polypeptide is partially deleted, and wherein the attL DNA sequence and the attR DNA sequence are completely deleted.

41. The genetically engineered bacterium of claim 34, wherein the nucleic acid sequence encoding the recombinase polypeptide and the attL DNA sequence are partially deleted, and wherein the attR DNA sequence is completely deleted.

42. The genetically engineered bacterium of claim 34, wherein the nucleic acid sequence encoding the recombinase polypeptide and the attR DNA sequence are partially deleted, and wherein the attL DNA sequence is completely deleted.

43. The genetically engineered bacterium of claim 34, wherein the recombinase polypeptide encoded by the modified nucleic acid sequence, the modified attL DNA sequence, and the modified attR DNA sequence are inactive.

44. The genetically engineered bacterium of claim 34, wherein the nucleic acid sequence encoding the recombinase polypeptide, if present, comprises a xerC2 gene or a portion or homolog thereof, a xerC3 gene or a portion or homolog thereof, or a xerD gene or a portion or homolog thereof.

45. The genetically engineered bacterium of any one of claims 34-44, wherein the bacterium is a gram-negative bacterium.

46. The genetically engineered bacterium of any one of claims 34-44, wherein the bacterium is a gram-positive bacterium.

47. The genetically engineered bacterium of any one of claims 34-46, wherein the bacterium is a nitrogen-fixing bacterium.Attorney Docket No. PIV-OOO53 WO48. The genetically engineered bacterium of claim 47, wherein the nitrogen-fixing bacterium is of a genus selected from Azospirillum, Kosakonia, Klebsiella, and Paraburkholderia.

49. The genetically engineered bacterium of claim 48, wherein the nitrogen-fixing bacterium is of the species Kosakonia oryzendophytica or Kosakonia arachidis.

50. The genetically engineered bacterium of claim 48, wherein the nitrogen-fixing bacterium is of the species Kosakonia oryzendophytica, strain 5812 (ATCC Accession No. PTA-127355).

51. The genetically engineered bacterium of claim 48, wherein the nitrogen-fixing bacterium further comprises modifications in genes involved in nitrogen fixation or nitrogen assimilation.

52. The genetically engineered bacterium of any one of claims 34-51, wherein the modified prophage has a DNA sequence that is at least 90% identical to SEQ ID NO: 9-11.

53. The genetically engineered bacterium of any one of claims 34-52, wherein the bacterium having the modified prophage has an increased competitive fitness when co-cultured with a corresponding bacterial cell that does not have the modified prophage.

54. The genetically engineered bacterium of claim 53, wherein the increased competitive fitness is determined under an environmental stress condition.

55. The genetically engineered bacterium of claim 54, wherein the environmental stress condition is selected from the group consisting of nutrient stress, salt stress, osmotic stress, cold stress, heat stress, excess UV radiation, the presence of antibiotics, and DNA damage.

56. The genetically engineered bacterium of any one of claims 34-55, wherein the genetically engineered bacterium having the modified prophage has an increased cell growth rate in comparison to a corresponding bacterium that does not have the modified prophage.Attorney Docket No. PIV-OOO53 WO57. The genetically engineered bacterium of claim 56, wherein the growth rate is measured in a liquid culture medium, in a soil sample, or in a field.

58. The genetically engineered bacterium of any of claims 34-57, wherein the genetically engineered bacterium having the modified prophage does not contain an activatible prophage.

59. The genetically engineered bacterium of any of claims 34-58, wherein all of the phage recombinase sequences that are present in the bacterial genome are inactivated or removed.

60. A modified prophage having a DNA sequence that is at least 90% identical to SEQ. ID NO: 9-11 and cannot be self-excised from a host bacterial cell.

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