LOCUS       Exported               14031 bp DNA     circular SYN 03-DEC-2024
DEFINITION  Single-plasmid V. cholerae INTEGRATE system, encodes all proteins, 
            crRNA, and donor DNA. Entry vector encodes non-targeting crRNA, with
            BsaI sites for spacer cloning. pBBR1 backbone..
ACCESSION   .
VERSION     .
KEYWORDS    pSL1142
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14031)
  AUTHORS   Vo PLH, Ronda C, Klompe SE, Chen EE, Acree C, Wang HH, Sternberg SH
  TITLE     CRISPR RNA-guided integrases for high-efficiency, multiplexed 
            bacterial genome engineering.
  JOURNAL   Nat Biotechnol. 2020 Nov 23. pii: 10.1038/s41587-020-00745-y. doi: 
            10.1038/s41587-020-00745-y.
  PUBMED    33230293
REFERENCE   2  (bases 1 to 14031)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 14031)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 14031)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2020 Nov 23. pii: 10.1038/s41587-020-00745-y. doi: 
            10.1038/s41587-020-00745-y."
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..14031
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(122..893)
                     /direction=LEFT
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     misc_feature    998..1049
                     /label=oriT
     CDS             1117..2115
                     /codon_start=1
                     /product="MobV family relaxase"
                     /label=Mob
                     /translation="MAAYAIMRCKKLAKMGNVAASLKHAYRERETPNADASRTPENEHW
                     AASSTDEAMGRLRELLPEKRRKDAVLAVEYVMTASPEWWKSASQEQQAAFFEKAHKWLA
                     DKYGADRIVTASIHRDETSPHMTAFVVPLTQDGRLSAKEFIGNKAQMTRDQTTFAAAVA
                     DLGLQRGIEGSKARHTRIQAFYEALERPPVGHVTISPQAVEPRAYAPQGLAEKLGISKR
                     VETPEAVADRLTKAVRQGYEPALQAAAGAREMRKKADQAQETARDLRERLKPVLDALGP
                     LNRDMQAKAAAIIKAVGEKLLTEQREVQRQKQAQRQQERGRAHFPEKCHLG"
     CDS             2373..3164
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        3639..3673
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological
                     Parts BBa_J23119)"
     CDS             3816..5000
                     /codon_start=1
                     /label=TniQ
                     /note="TniQ family protein [Vibrio cholerae]"
                     /translation="MFLQRPKPYSDESLESFFIRVANKNGYGDVHRFLEATKRFLQDID
                     HNGYQTFPTDITRINPYSAKNSSSARTASFLKLAQLTFNEPPELLGLAINRTNMKYSPS
                     TSAVVRGAEVFPRSLLRTHSIPCCPLCLRENGYASYLWHFQGYEYCHSHNVPLITTCSC
                     GKEFDYRVSGLKGICCKCKEPITLTSRENGHEAACTVSNWLAGHESKPLPNLPKSYRWG
                     LVHWWMGIKDSEFDHFSFVQFFSNWPRSFHSIIEDEVEFNLEHAVVSTSELRLKDLLGR
                     LFFGSIRLPERNLQHNIILGELLCYLENRLWQDKGLIANLKMNALEATVMLNCSLDQIA
                     SMVEQRILKPNRKSKPNSPLDVTDYLFHFGDIFCLWLAEFQSDEFNRSFYVSRW"
     CDS             5001..6923
                     /codon_start=1
                     /label=Cas8
                     /note="Cas8 from Vibrio vulnificus, type I-F 
                     CRISPR-associated protein Csy2"
                     /translation="MQTLKELIASNPDDLTTELKRAFRPLTPHIAIDGNELDALTILVN
                     LTDKTDDQKDLLDRAKCKQKLRDEKWWASCINCVNYRQSHNPKFPDIRSEGVIRTQALG
                     ELPSFLLSSSKIPPYHWSYSHDSKYVNKSAFLTNEFCWDGEISCLGELLKDADHPLWNT
                     LKKLGCSQKTCKAMAKQLADITLTTINVTLAPNYLTQISLPDSDTSYISLSPVASLSMQ
                     SHFHQRLQDENRHSAITRFSRTTNMGVTAMTCGGAFRMLKSGAKFSSPPHHRLNSKRSW
                     LTSEHVQSLKQYQRLNKSLIPENSRIALRRKYKIELQNMVRSWFAMQDHTLDSNILIQH
                     LNHDLSYLGATKRFAYDPAMTKLFTELLKRELSNSINNGEQHTNGSFLVLPNIRVCGAT
                     ALSSPVTVGIPSLTAFFGFVHAFERNINRTTSSFRVESFAICVHQLHVEKRGLTAEFVE
                     KGDGTISAPATRDDWQCDVVFSLILNTNFAQHIDQDTLVTSLPKRLARGSAKIAIDDFK
                     HINSFSTLETAIESLPIEAGRWLSLYAQSNNNLSDLLAAMTEDHQLMASCVGYHLLEEP
                     KDKPNSLRGYKHAIAECIIGLINSITFSSETDPNTIFWSLKNYQNYLVVQPRSINDETT
                     DKSSL"
     CDS             6895..7953
                     /codon_start=1
                     /label=Cas7
                     /note="type I-F CRISPR-associated protein Cas7 [Vibrio 
                     cholerae]"
                     /translation="MKLPTNLAYERSIDPSDVCFFVVWPDDRKTPLTYNSRTLLGQMEA
                     ASLAYDVSGQPIKSATAEALAQGNPHQVDFCHVPYGASHIECSFSVSFSSELRQPYKCN
                     SSKVKQTLVQLVELYETKIGWTELATRYLMNICNGKWLWKNTRKAYCWNIVLTPWPWNG
                     EKVGFEDIRTNYTSRQDFKNNKNWSAIVEMIKTAFSSTDGLAIFEVRATLHLPTNAMVR
                     PSQVFTEKESGSKSKSKTQNSRVFQSTTIDGERSPILGAFKTGAAIATIDDWYPEATEP
                     LRVGRFGVHREDVTCYRHPSTGKDFFSILQQAEHYIEVLSANKTPAQETINDMHFLMAN
                     LIKGGMFQHKGD"
     CDS             7956..8555
                     /codon_start=1
                     /label=Cas6
                     /note="type I-F CRISPR-associated endoribonuclease 
                     Cas6/Csy4 [Vibrio cholerae]"
                     /translation="MKWYYKTITFLPELCNNESLAAKCLRVLHGFNYQYETRNIGVSFP
                     LWCDATVGKKISFVSKNKIELDLLLKQHYFVQMEQLQYFHISNTVLVPEDCTYVSFRRC
                     QSIDKLTAAGLARKIRRLEKRALSRGEQFDPSSFAQKEHTAIAHYHSLGESSKQTNRNF
                     RLNIRMLSEQPREGNSIFSSYGLSNSENSFQPVPLI"
     CDS             8577..9269
                     /codon_start=1
                     /label=TnsA
                     /note="TnsA endonuclease N-terminal domain-containing 
                     protein [Vibrio]"
                     /translation="MATSLPTPSAITTSALEYAFHTPARNLTKSRGKNIHRYVSVKMSK
                     RITVESTLECDACYHFDFEPSIVRFCAQPIRFLYYLNGQSHSYVPDFLVQFDTNEFVLY
                     EVKSAYAKNKPDFDVEWEAKVKAATELGLELELVEESDIRDTVVLNNLKRMHRYASKDE
                     LNNVHNSLLKIIKYNGAQSARCLGEQLGLKGRTVLPILCDLLSRCLLDTRLDKPLSLES
                     RFELASYG"
     CDS             9262..11073
                     /codon_start=1
                     /label=TnsB
                     /translation="MAKKGFSSFHRKAVSSQDTLESIELVSSANCLESVTYQDISAFPE
                     TIAVEINFRLSILRFLARKCETIVAKSIEPHRVELQQNYSRKIPSAITIYRWWLAFRKS
                     DYNPISLAPNIKDRGNRETKVSTVVDSIMEQAVERVISGRKVNVSSAYKRVRRKVRQYN
                     LTHGTKYTYPKYESVRKRVKKKTPFELLAAGKGERVAKREFRRMGKKILTSSVLERVEI
                     DHTVVDLFAVHEEYRIPLGRPWLTQLVDCYSKAVIGFYLGFEPPSYVSVSLALKNAIQR
                     KDDLISSYESIENEWLCYGIPDLLVTDNGKEFLSKAFDQACESLLINVHQNKVETPDNK
                     PHVERNYGTINTSLLDDLPGKSFSQYLQREGYDSVGEATLTLNEIREIYLIWLVDIYHK
                     KPNQRGTNCPNVAWKKGCQEWEPEEFSGSKDELDFKFAIVDYKQLTKVGITVYKELSYS
                     NDRLAEYRGKKGNHKVQFKYNPECMAVIWVLDEDMNEYFTVNAIDYEYASRVSLWQHKY
                     NMKYQAELNSAEYDEDKEIDAEIKIEEIADRSIVKTNKIRARRRGARHQENSARAKSIS
                     NANPASIQKHEDEIVSADNDDWDIDYV"
     CDS             10655..10666
                     /codon_start=1
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
                     /translation="IEGR"
     CDS             11085..12077
                     /codon_start=1
                     /label=TnsC
                     /note="Tn6677 Vibrio cholerae transposon TnsC (VchTnsC) 
                     [Vibrio cholerae]"
                     /translation="MSETREARISRAKRAFVSTPSVRKILSYMDRCRDLSDLESEPTCM
                     MVYGASGVGKTTVIKKYLNQNRRESEAGGDIIPVLHIELPDNAKPVDAARELLVEMGDP
                     LALYETDLARLTKRLTELIPAVGVKLIIIDEFQHLVEERSNRVLTQVGNWLKMILNKTK
                     CPIVIFGMPYSKVVLQANSQLHGRFSIQVELRPFSYQGGRGVFKTFLEYLDKALPFEKQ
                     AGLANESLQKKLYAFSQGNMRSLRNLIYQASIEAIDNQHETITEEDFVFASKLTSGDKP
                     NSWKNPFEEGVEVTEDMLRPPPKDIGWEDYLRHSTPRVSKPGRNKNFFE"
     terminator      12115..12162
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    12183..12307
                     /label=VchINT R-end
     CDS             12317..12973
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     misc_feature    complement(13013..13157)
                     /label=VchINT L-end
     CDS             complement(join(13493..14031,1..121))
                     /codon_start=1
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
ORIGIN
        1 ccgccgtggg tttcctcgca ataagcccgg cccacgcctc atgcgctttg cgttccgttt
       61 gcacccagtg accgggcttg ttcttggctt gaatgccgat ttctctggac tgcgtggcca
      121 tgcttatctc catgcggtag gggtgccgca cggttgcggc accatgcgca atcagctgca
      181 acttttcggc agcgcgacaa caattatgcg ttgcgtaaaa gtggcagtca attacagatt
      241 ttctttaacc tacgcaatga gctattgcgg ggggtgccgc aatgagctgt tgcgtacccc
      301 ccttttttaa gttgttgatt tttaagtctt tcgcatttcg ccctatatct agttctttgg
      361 tgcccaaaga agggcacccc tgcggggttc ccccacgcct tcggcgcggc tccccctccg
      421 gcaaaaagtg gcccctccgg ggcttgttga tcgactgcgc ggccttcggc cttgcccaag
      481 gtggcgctgc ccccttggaa cccccgcact cgccgccgtg aggctcgggg ggcaggcggg
      541 cgggcttcgc ccttcgactg cccccactcg cataggcttg ggtcgttcca ggcgcgtcaa
      601 ggccaagccg ctgcgcggtc gctgcgcgag ccttgacccg ccttccactt ggtgtccaac
      661 cggcaagcga agcgcgcagg ccgcaggccg gaggcttttc cccagagaaa attaaaaaaa
      721 ttgatggggc aaggccgcag gccgcgcagt tggagccggt gggtatgtgg tcgaaggctg
      781 ggtagccggt gggcaatccc tgtggtcaag ctcgtgggca ggcgcagcct gtccatcagc
      841 ttgtccagca gggttgtcca cgggccgagc gaagcgagcc agccggtggc cgctcgcggc
      901 catcgtccac atatccacgg gctggcaagg gagcgcagcg accgcgcagg gcgaagcccg
      961 gagagcaagc ccgtagggcg ccgcagccgc cgtaggcggt cacgactttg cgaagcaaag
     1021 tctagtgagt atactcaagc attgagtggc ccgccggagg caccgccttg cgctgccccc
     1081 gtcgagccgg ttggacacca aaagggaggg gcaggcatgg cggcatacgc gatcatgcga
     1141 tgcaagaagc tggcgaaaat gggcaacgtg gcggccagtc tcaagcacgc ctaccgcgag
     1201 cgcgagacgc ccaacgctga cgccagcagg acgccagaga acgagcactg ggcggccagc
     1261 agcaccgatg aagcgatggg ccgactgcgc gagttgctgc cagagaagcg gcgcaaggac
     1321 gctgtgttgg cggtcgagta cgtcatgacg gccagcccgg aatggtggaa gtcggccagc
     1381 caagaacagc aggcggcgtt cttcgagaag gcgcacaagt ggctggcgga caagtacggg
     1441 gcggatcgca tcgtgacggc cagcatccac cgtgacgaaa ccagcccgca catgaccgcg
     1501 ttcgtggtgc cgctgacgca ggacggcagg ctgtcggcca aggagttcat cggcaacaaa
     1561 gcgcagatga cccgcgacca gaccacgttt gcggccgctg tggccgatct agggctgcaa
     1621 cggggcatcg agggcagcaa ggcacgtcac acgcgcattc aggcgttcta cgaggccctg
     1681 gagcggccac cagtgggcca cgtcaccatc agcccgcaag cggtcgagcc acgcgcctat
     1741 gcaccgcagg gattggccga aaagctggga atctcaaagc gcgttgagac gccggaagcc
     1801 gtggccgacc ggctgacaaa agcggttcgg caggggtatg agcctgccct acaggccgcc
     1861 gcaggagcgc gtgagatgcg caagaaggcc gatcaagccc aagagacggc ccgtgacctt
     1921 cgggagcgcc tgaagcccgt tctggacgcc ctggggccgt tgaatcggga tatgcaggcc
     1981 aaggccgccg cgatcatcaa ggccgtgggc gaaaagctgc tgacggaaca gcgggaagtc
     2041 cagcgccaga aacaggccca gcgccagcag gaacgcgggc gcgcacattt ccccgaaaag
     2101 tgccacctgg gatgaatgtc agctactggg ctatctggac aagggaaaac gcaagcgcaa
     2161 agagaaagca ggtagcttgc agtgggctta catggcgata gctagactgg gcggttttat
     2221 ggacagcaag cgaaccggaa ttgccagctg gggcgccctc tggtaaggtt gggaagccct
     2281 gcaaagtaaa ctggatggct ttcttgccgc caaggatctg atggcgcagg ggatcaagat
     2341 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     2401 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     2461 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     2521 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     2581 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     2641 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     2701 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     2761 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     2821 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     2881 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     2941 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     3001 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     3061 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     3121 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     3181 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     3241 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     3301 ccagcgcggg gatctcatgc tggagttctt cgcccacccc catgggcaaa tattatacgc
     3361 aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct
     3421 tccatgtcgg cagaatgctt aatgaattac aacagttttt atgcatgcgc ccaatacgca
     3481 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     3541 actggtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     3601 gaccgagcgc agcgagtcag tgagcgagga cacaagcttt gacagctagc tcagtcctag
     3661 gtataatact agcgtcgacg tggagatata ccatgggtga actgccgagt aggtagctga
     3721 taacgagacc tctgtcttgt cagctagggt ggtctcgtga actgccgagt aggtagctga
     3781 taacggatcc gaattcgagc gaaggagata tacatatgtt tttgcaaaga cctaaacctt
     3841 acagcgatga aagtttagaa agtttcttta tccgagtggc taacaaaaat ggctacggtg
     3901 atgtccatcg cttcctagaa gccactaaac gattccttca agacattgac cataatggct
     3961 atcaaacctt tccgactgat ataactcgga taaacccata ctcagctaaa aacagttcca
     4021 gcgcacgaac tgcgtcattc ctgaaacttg cacaattgac atttaatgaa ccgccagagc
     4081 tacttgggtt ggcaattaac agaacaaaca tgaaatactc gccgtcaact agcgcggttg
     4141 ttagaggtgc agaagtcttt cctcgcagtt tactacggac gcactccatc ccctgctgtc
     4201 ctttgtgtct gcgagaaaat ggctacgcct cctacctttg gcactttcag gggtacgaat
     4261 actgccacag ccataacgta cctttaatta ccacttgtag ctgtggtaag gagtttgact
     4321 accgagtatc tgggttaaag ggcatttgct gcaaatgcaa ggagcctatc accttaacca
     4381 gcagggagaa cggtcatgag gcagcgtgta ctgtttcaaa ctggcttgct ggccatgaat
     4441 ctaaacctct gccaaatctt cctaaaagct accgatgggg tttagttcat tggtggatgg
     4501 gtattaaaga tagcgagttc gatcactttt cgttcgttca atttttctca aactggccaa
     4561 ggtcattcca ctcgataatc gaagatgaag tagagttcaa ccttgagcat gctgttgtca
     4621 gcacgtctga attacgacta aaagatcttc ttggtcgatt gtttttcggt tcaattcggt
     4681 tacctgagcg gaatcttcaa cacaatatca tccttggtga gcttctctgc tatttagaaa
     4741 atcgtttatg gcaagacaag ggattaatcg ccaacctcaa aatgaacgcg ttagaggcga
     4801 ctgtaatgtt aaattgtagc ctcgatcaga ttgcatcaat ggttgaacaa cgcatcttga
     4861 agccaaatcg aaaaagcaag cccaacagcc ctcttgatgt taccgattat ctatttcatt
     4921 tcggcgatat tttctgtctt tggttagctg agttccaaag cgatgagttt aaccgttcgt
     4981 tttatgtgtc gaggtggtaa atgcaaactc tgaaagaact aatcgcatcc aatcctgacg
     5041 acttaacaac tgaacttaag agagcatttc gtccactcac accgcatatt gcaattgatg
     5101 gtaatgaact tgacgcactg acgatattag tcaatttaac cgataagact gatgatcaga
     5161 aagacctgct cgatcgagcc aaatgcaagc aaaaacttcg agatgaaaaa tggtgggcta
     5221 gctgcataaa ttgcgttaat tacagacaaa gccataaccc aaaattcccg gatatacgtt
     5281 ctgaaggcgt gattcgaacc caagccctgg gtgaattacc gagtttccta ctctcgtcct
     5341 caaaaatccc accataccat tggtcatata gccatgattc aaaatacgtc aacaaaagcg
     5401 cattcctcac caatgagttt tgttgggatg gtgagatctc atgtttgggt gagcttctta
     5461 aagatgcaga tcacccactt tggaatactt tgaaaaagtt aggttgttct caaaaaactt
     5521 gcaaagcaat ggcaaaacaa ctagctgata ttactctcac gactatcaat gtcacactcg
     5581 caccaaatta cctgactcaa atatctctcc ccgatagtga tacatcctac atttcactct
     5641 ctcctgttgc atcgctatcg atgcaaagcc actttcatca gaggctccaa gatgaaaatc
     5701 ggcatagcgc gataacgcgg tttagccgaa ctaccaacat gggggtcact gcgatgacat
     5761 gtggcggcgc atttagaatg ttaaagtctg gcgctaagtt ttccagcccc cctcatcacc
     5821 gattaaacag taaacgaagt tggttgacgt cagagcatgt tcagtcatta aaacagtacc
     5881 agcgcctcaa caaaagcctc atacctgaaa actctcggat tgcactccgt agaaaataca
     5941 aaatcgagct tcaaaatatg gtcagatctt ggtttgcaat gcaagaccat acccttgatt
     6001 cgaacatact tatccaacat ttgaatcatg acctatctta cttaggagcc acaaaacgtt
     6061 ttgcatacga tccagcgatg accaagctct ttactgagct tttgaaacga gagttatcaa
     6121 attcaatcaa taatggtgag caacacacta atggatcgtt tttagtccta ccgaatatca
     6181 gggtttgtgg cgcaacagct ttaagctccc cggtaacggt ggggattcca tcacttacag
     6241 ctttctttgg cttcgttcac gcatttgaac ggaatataaa tcgcaccacc tcatcgtttc
     6301 gtgttgaatc ctttgcgata tgcgtccatc aactacatgt cgagaagcga ggtttgacag
     6361 cagagtttgt ggaaaaaggc gacgggacta tatccgctcc cgcgacccgg gatgactggc
     6421 agtgtgatgt cgtatttagc cttattttga acaccaactt tgctcaacat attgaccaag
     6481 atacgttagt tacatcacta ccaaagcgat tggctcgggg ttcagcaaaa attgcgattg
     6541 atgactttaa acatatcaac tcattctcga cattagaaac agcgatcgaa tctctgccaa
     6601 tagaagctgg taggtggtta tcactttacg cacagtcaaa caataatcta agtgatctat
     6661 tagcagccat gacagaggac catcagctca tggcaagctg cgtcggttac cacttgttag
     6721 aagagcccaa agataaacca aactccctca gaggttacaa acacgctatc gccgagtgca
     6781 tcattggact cattaactca atcaccttta gctcagagac tgatcccaac acaatctttt
     6841 ggtcgctaaa gaactatcaa aactacctag tggtacagcc aaggagtatc aacgatgaaa
     6901 ctaccgacaa atctagccta tgagcgctct atcgacccat cagatgtctg tttttttgtc
     6961 gtctggcccg atgatagaaa aacaccttta acctacaatt ctcgtactct gctcgggcaa
     7021 atggaagcgg catcattagc ctatgatgtc tcaggtcaac caataaaaag tgccaccgct
     7081 gaggcgttag ctcaagggaa ccctcatcaa gttgatttct gccacgttcc atacggtgcg
     7141 agtcatattg aatgcagttt ctccgtctcg ttttcttctg aactacgtca accatataag
     7201 tgtaactcaa gcaaagttaa acaaacgcta gtgcaattag tcgagctcta cgaaacgaaa
     7261 atcggctgga ctgagctagc aacccgatat ttgatgaaca tttgcaacgg taaatggctg
     7321 tggaaaaata cccgtaaagc ctattgctgg aacattgtac ttacaccttg gccttggaac
     7381 ggggaaaagg ttggatttga agatattcgt actaactaca cctcacggca agactttaaa
     7441 aataataaaa attggtctgc tatagttgaa atgatcaaaa ccgcattttc tagtactgat
     7501 gggctggcga tatttgaagt cagggccacc ttgcacttgc caacgaatgc tatggtgcgg
     7561 ccaagccaag ttttcacaga aaaagaaagt ggcagtaaaa gtaaatctaa aactcaaaac
     7621 agtcgagttt ttcagagtac aactattgat ggtgaacgat cgccaatact aggggccttt
     7681 aaaacgggag cagctattgc aaccattgac gactggtatc ctgaagccac tgagccacta
     7741 agggtcggac ggtttggggt tcatcgcgaa gatgtcactt gctaccgtca tccgtctacc
     7801 ggaaaagatt ttttctcgat attacaacaa gcagagcact atattgaagt gttgagcgcc
     7861 aacaaaactc ccgctcaaga aactatcaac gacatgcact ttttaatggc taacctgatt
     7921 aagggtggga tgttccagca taaaggagac tgactgtgaa atggtattat aagacaatca
     7981 cctttctgcc agagttgtgc aacaacgagt cactggctgc aaagtgtctc cgcgttctgc
     8041 atggatttaa ctatcagtat gagacacgaa atatcggcgt ttcatttccg ctttggtgtg
     8101 atgcaacggt tggaaaaaag atttcatttg tcagcaagaa caagatagaa ctcgacttac
     8161 tacttaaaca acactatttc gtccaaatgg aacaacttca atactttcat atatccaaca
     8221 ctgttctcgt cccagaagat tgtacatacg tttcctttag acgctgtcaa tctatagata
     8281 agctcacagc agcagggctg gcaaggaaaa tcagacgcct ggagaaacgt gctctatcta
     8341 gaggcgagca atttgaccca tcatcttttg ctcaaaaaga gcatactgca atagcgcact
     8401 accactcact tggggagtcc agcaaacaga cgaaccgcaa ctttcgactc aatatcagga
     8461 tgctctcgga gcaaccccgt gaggggaact cgatttttag tagctatggc ttatcaaatt
     8521 cagaaaactc gtttcagcct gtacccttaa tctgaccttt aataaggaga tataccatgg
     8581 cgacaagttt acctacgccc tcagcaatta cgacttcggc gttagagtat gcattccata
     8641 ctcccgctcg caatctaacg aaatctcgcg gaaaaaacat tcatcgttat gtcagtgtaa
     8701 agatgagtaa gaggattacg gtagaatcta ctctagagtg tgatgcctgc tatcactttg
     8761 attttgagcc aagtattgtt cgcttttgcg ctcaaccgat tcgattttta tattatctca
     8821 atggtcagtc tcactcctat gttcctgact ttctagttca atttgatacc aacgagtttg
     8881 ttctatatga agtaaagtca gcttatgcta agaacaaacc tgattttgat gttgaatggg
     8941 aggcgaaagt aaaagcagca actgaactag ggctagaatt ggagcttgtt gaagagagtg
     9001 atattaggga tacggttgta ttaaataatc ttaagcgcat gcatcgttat gcttcgaaag
     9061 atgagctgaa taacgtacat aactctctct taaaaataat aaagtacaat ggcgcccaat
     9121 ctgcaagatg cttgggagaa cagttgggtt taaaaggccg aactgtttta ccaattttgt
     9181 gcgatttgct gtcaaggtgt ttactcgata cacgtttgga taagcctcta tctcttgaat
     9241 ctcgatttga gttggccagt tatggctaag aaagggttct caagtttcca tagaaaagca
     9301 gtctcttccc aagatacgct tgaatcgata gagcttgtct ctagcgctaa ttgtctagaa
     9361 agtgttacgt atcaagatat atcagcattt cccgaaacaa ttgcggtaga gattaatttc
     9421 cgattaagca ttcttcgctt tttagcgcga aagtgcgaaa ccattgtggc caaatcaatt
     9481 gaaccacatc gtgtagagct acagcaaaac tatagtagaa aaatacccag tgcaataacg
     9541 atatatcgat ggtggcttgc ttttcgaaaa tcagactaca accccattag cttagcacct
     9601 aatatcaagg atagaggtaa tagagagaca aaagtgtcaa cagttgttga ttctattatg
     9661 gaacaggcag ttgaaagagt tatatctgga cgaaaagtca atgttagctc tgcatataaa
     9721 cgtgttcgac gaaaagttcg tcaatacaat ctaactcatg gaacgaaata cacgtatcct
     9781 aagtacgaat ctgtaagaaa gcgagtaaaa aagaaaaccc catttgagtt attagccgca
     9841 gggaaagggg agagagtagc taagagagag tttcgccgaa tgggaaaaaa gatcctcacg
     9901 tctagcgtgc tagagagggt tgaaatagat cacactgtcg ttgacctttt tgcagtacac
     9961 gaagagtatc gaatcccatt gggccgacct tggcttactc aattggttga ttgttacagt
    10021 aaagctgtaa tcggttttta tttaggtttc gagcctccta gctatgtgtc ggtttccctt
    10081 gcacttaaga atgcaataca acgcaaagat gacttaatct cctcgtatga atcgatcgag
    10141 aatgaatggc tatgttatgg catcccagac ctactcgtaa ctgataatgg taaagagttt
    10201 ttgtcgaaag catttgatca agcatgtgaa tcactattga tcaatgtgca tcaaaataaa
    10261 gttgagacgc ccgacaacaa acctcatgtt gaacgtaact acgggactat taatacttct
    10321 ctgttagacg atttacctgg gaaatccttc agccagtacc ttcaaagaga agggtacgac
    10381 tctgtgggag aagctaccct tacactcaat gagattagag aaatttactt aatttggttg
    10441 gtggatattt atcataaaaa acccaatcag agaggcacta attgtcctaa tgttgcgtgg
    10501 aaaaagggtt gtcaagaatg ggaaccagag gagttctctg gttctaaaga cgaattagac
    10561 tttaaatttg ctattgttga ttacaaacaa cttactaaag tagggataac tgtctacaaa
    10621 gaactgagtt atagcaatga ccgtttagct gaatatagag ggaagaaagg aaaccataaa
    10681 gttcagttca agtataaccc tgagtgtatg gcagttattt gggtgttgga tgaggatatg
    10741 aatgagtact ttacagttaa tgcgattgac tacgaatatg caagtagagt atcactttgg
    10801 caacataaat ataacatgaa atatcaagca gaactaaatt cagcagaata tgatgaggac
    10861 aaggaaattg atgcagaaat aaaaattgaa gaaatcgcag atcgttcaat tgttaagact
    10921 aacaaaatca gagctcggag gcgtggcgct aggcatcaag agaatagcgc aagggctaag
    10981 tcaatcagta atgcgaaccc ggcctcgata caaaaacatg aagatgaaat cgttagtgca
    11041 gataatgacg attgggatat tgattatgtc tgagaaatcg tcaaatgagt gaaacgcgtg
    11101 aggctcgtat atcaagagct aaaagggcat ttgtatccac accgagcgtt aggaaaatct
    11161 tgagttacat ggatagatgt agagatctat cagacctaga gtctgagcct acatgcatga
    11221 tggtctatgg tgcttcgggt gtaggtaaaa cgaccgtcat caagaaatac ttaaatcaaa
    11281 acagaagaga gtccgaagcc gggggcgata taataccggt tttgcatatt gagttgccag
    11341 acaatgcgaa gccagtagat gcagcaaggg aattgctggt tgaaatgggt gacccgctag
    11401 cactttatga aactgactta gctagattga cgaaaagact gactgaatta atccctgcgg
    11461 tcggcgtgaa gctgattatt atcgatgagt tccaacattt ggtggaagaa aggtcaaatc
    11521 gggttcttac ccaagtaggt aattggctaa aaatgatact taacaaaacg aaatgtccaa
    11581 ttgttatatt tggtatgcca tactcaaaag ttgtactgca agcaaactcg caacttcacg
    11641 ggcgattttc cattcaggtt gaactgcgcc cctttagcta ccagggaggt agaggtgtat
    11701 ttaaaacttt tttggaatac cttgataaag ccctaccttt tgaaaaacag gctggcttag
    11761 ccaacgaaag tttgcagaaa aaattgtatg cattctctca gggaaacatg cgttcgttga
    11821 gaaaccttat ttatcaagca tctatcgaag caattgataa tcagcatgag acgataaccg
    11881 aagaagattt cgtttttgca tcgaagttga catcgggcga taaacccaac tcatggaaaa
    11941 atccttttga ggagggtgtt gaggtaacag aagatatgtt acgaccgcca ccaaaagata
    12001 ttggttggga ggactatttg agacattcaa ccccgagagt gagtaaacca ggtagaaata
    12061 aaaacttttt cgaataacct aggctgctgc gctgctgcca ccgctgagca ataactagca
    12121 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaacc tcaggcattt
    12181 gttgttgata caaccataaa atgataatta cacccataaa ttgataatta tcacacccat
    12241 aaattgatat tgcctcttca tggtctaaac ttcagtaagt ttacgacatt ttcctcgagg
    12301 tcatttccgg ggatccatgg agaaaaaaat cactggatat accaccgttg atatatccca
    12361 atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca
    12421 gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt
    12481 ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg agttccgtat
    12541 ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt
    12601 ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca
    12661 gtttctacac atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc
    12721 taaagggttt attgagaata tgtttttcgt ctcagccaat ccctgggtga gtttcaccag
    12781 ttttgattta aacgtggcca atatggacaa cttcttcgcc cccgttttca ctatgggcaa
    12841 atattatacg caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt
    12901 ttgtgatggc ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg
    12961 gcagggcggg gcgtaatttt tttaaggcag ttattggtgc ccttctagag tccttactgc
    13021 agtagttttg ctgaaatact cgattcacaa aaatatcaac ttatggttgt tttgtgagat
    13081 atcaatatat ggttgttttg tggttaagtt gctgattata aataattatt aaatatcact
    13141 ttatggttgc atcaacaggt accgagctcg aattcactgg ccgtcgtttt acaacgtcgt
    13201 gactgggaaa accctggcgt tacccaactt aatcgccttg cggccagttt gctcaggctc
    13261 tccccgtgga ggtaataatt gacgatatga tcatttattc tgcctcccag agcctgataa
    13321 aaacggtgaa tccgttagcg aggtgccgcc ggcttccatt caggtcgagg tggcccggct
    13381 ccatgcaccg cgacgcaacg cggggaggca gacaaggtat agggcggcga ggcggctaca
    13441 gccgatagtc tggaacagcg cacttacggg ttgctgcgca acccaagtgc taccggcgcg
    13501 gcagcgtgac ccgtgtcggc ggctccaacg gctcgccatc gtccagaaaa cacggctcat
    13561 cgggcatcgg caggcgctgc tgcccgcgcc gttcccattc ctccgtttcg gtcaaggctg
    13621 gcaggtctgg ttccatgccc ggaatgccgg gctggctggg cggctcctcg ccggggccgg
    13681 tcggtagttg ctgctcgccc ggatacaggg tcgggatgcg gcgcaggtcg ccatgcccca
    13741 acagcgattc gtcctggtcg tcgtgatcaa ccaccacggc ggcactgaac accgacaggc
    13801 gcaactggtc gcggggctgg ccccacgcca cgcggtcatt gaccacgtag gccgacacgg
    13861 tgccggggcc gttgagcttc acgacggaga tccagcgctc ggccaccaag tccttgactg
    13921 cgtattggac cgtccgcaaa gaacgtccga tgagcttgga aagtgtcttc tggctgacca
    13981 ccacggcgtt ctggtggccc atctgcgcca cgaggtgatg cagcagcatt g
//
