LOCUS       sequence-375864-       17515 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multi-host shuttle vector that can be transferred by conjugation and
            replicate in S. meliloti, E. coli, S. cerevisiae, and P. 
            tricornutum. Contains S. meliloti pSymA origin and Nm/Kan 
            resistance..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17515)
  AUTHORS   Brumwell SL, MacLeod MR, Huang T, Cochrane RR, Meaney RS, Zamani M, 
            Matysiakiewicz O, Dan KN, Janakirama P, Edgell DR, Charles TC, Finan
            TM, Karas BJ
  TITLE     Designer Sinorhizobium meliloti strains and multi-functional vectors
            enable direct inter-kingdom DNA transfer.
  JOURNAL   PLoS One. 2019 Jun 17;14(6):e0206781. doi: 
            10.1371/journal.pone.0206781. eCollection 2019.
  PUBMED    31206509
REFERENCE   2  (bases 1 to 17515)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
            date: "2019-06-17"; volume: "14(6):e0206781. doi"; pages: " 
            10.1371/journal.pone.0206781. eCollection 2019"
COMMENT     Sequence Label: pAGE3.0
FEATURES             Location/Qualifiers
     source          1..17515
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             600..1169
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGFT
                     LREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNRR
                     LTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFTL
                     CGLDTALYDGTASDGEQALYMSMPCP"
     rep_origin      5904..6518
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     rep_origin      5995..6526
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      6594..6813
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             6904..7659
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     misc_feature    7662..7912
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     misc_feature    8074..8577
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        8578..8764
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             8765..9427
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     CDS             9617..10792
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     CDS             10792..11763
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     misc_feature    11836..12309
                     /gene="sopC"
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     misc_feature    12569..12967
                     /label=cos
                     /note="lambda cos site; allows packaging into phage lambda 
                     particles"
     protein_bind    complement(12985..13018)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     13086..13105
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(13185..13460)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSYLGETIEYSSLHACRSTLEDPRVPSSNSPYSESYYNSL
                     AVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRS"
     primer_bind     13298..13320
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     13312..13329
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     13313..13329
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     13336..13355
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        13336..13354
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    13357..13413
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(13456..13472)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13456..13472)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(13469..13491)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    13480..13496
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(13504..13510,13511..13528,13529..13534))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13549..13570
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(13790..14449)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     14364..14383
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(14450..14552)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             complement(15587..16381)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(15698..15717)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     16308..16327
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     oriT            16880..16989
                     /note="incP origin of transfer"
     CDS             17022..17393
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
ORIGIN
        1 atcgatacta gcttgattgg gatatctcgc tcgtgcttgt cgcgtgctat gtcttttagg
       61 gtactttgaa cctacgttcg tacttgtata atatgatcat cgtattatcg tttttcatcc
      121 gtccagcgca aaatgcatta gcagctagtc ctagcgtgcg gagctacctg gacaggtgca
      181 tgacggatgc gtgtccttca gtgactttct aattaacagt aacttcttta cttatgtttc
      241 agtttgtaag aagcgggatt cgctcgtcgg ttgacatctg attggactgc gtcggcacat
      301 gaaaactaca ttgtgaaatc tgctaaaact ccgggtatct ctgacacaaa acgattcggc
      361 ttcgcaattt caacattacg gtcaaggcta acgtatcttt ctcggtcaac ttcagattat
      421 gccgattaaa ttgtcgtagc tttcaaggcg ttttgagtac tgcggcagtt gttgaacctg
      481 caaggagaag atctcgacaa cagaataaag cgaaaaatgg gtctcatgca ctaacactca
      541 ggcctccctc ataatctctg tttgagttta ccaacaacac atatatacat ttcgacaaaa
      601 tgaccactct tgacgacacg gcttaccggt accgcaccag tgtcccgggg gacgccgagg
      661 ccatcgaggc actggatggg tccttcacca ccgacaccgt cttccgcgtc accgccaccg
      721 gggacggctt caccctgcgg gaggtgccgg tggacccgcc cctgaccaag gtgttccccg
      781 acgacgaatc ggacgacgaa tcggacgacg gggaggacgg cgacccggac tcccggacgt
      841 tcgtcgcgta cggggacgac ggcgacctgg cgggcttcgt ggtcgtctcg tactccggct
      901 ggaaccgccg gctgaccgtc gaggacatcg aggtcgcccc ggagcaccgg gggcacgggg
      961 tcgggcgcgc gttgatgggg ctcgcgacgg agttcgcccg cgagcggggc gccgggcacc
     1021 tctggctgga ggtcaccaac gtcaacgcac cggcgatcca cgcgtaccgg cggatggggt
     1081 tcaccctctg cggcctggac accgccctgt acgacggcac cgcctcggac ggcgagcagg
     1141 cgctctacat gagcatgccc tgcccctgat tttgttacat ttactgactt caaggagtcg
     1201 aggaatcgat actgccgtcg tttccaggat ccgaggtttc ataaactctg ttaacgttat
     1261 agaaacagac ttacctctcc tacgccattc acgtaatatt cgcaatatgc tattcttcct
     1321 ctgaagacca ggtttatgtg ctgcctgaaa ctatttcaat aagtcagctg cacttgcaca
     1381 gggtttcaca aggaaagcgt gtcttttttt ccaacgtagg cgtcgctttc gtctgactct
     1441 tactcttaca ttcacagcca atacttacaa ttagtaaaaa acctgtgctc gagagtgaaa
     1501 actccgtaac tataacggtc ctaaggtagc gaacactccg ttcgtggcgt cgcacgatcg
     1561 cgacatttga cgacgtgtgt tttgcctggg ctacgtatcg ggaatgtttg cgaaaccccc
     1621 gaaattgtac cgggatgtag gcacatcgct ggaaggtagc caggggggtc cggcgctgat
     1681 tacgaagtgc ggcgaccgac gaatttggag cacgcccgac gctcaggggt ggatgccgct
     1741 ggcgattccg cttgagtggg atcgctcggc acccgcgggt cggcatatgc ccgtgccgat
     1801 tgtgtcccca cggtgttggt caggcggccg gtatcacgac accccccgcc catcgaagcc
     1861 attcgctctc agcaacgcca tgagcatcgg gccaagcgag aatttccctt gccgcgcctt
     1921 ggcggtcaga tcgcgcagat agccgccagc agagttgata tgccctgccc tctctagaat
     1981 gcaggccacg gcgactccgg cgttttccgg tcccatgacc tcgcaggcct cctgataggc
     2041 cgaggggtaa acgccaagca tagatcgtac cgtgactgca gcgatcatga ggtcgcgcca
     2101 acttctcact gcaccgtcag gaccgtagtt catgatttcc gggcaggctt tcagtaccat
     2161 tcccagcgga aaggcccttt ttgccttgtt ggccggttca atggcttggc cgtctaaccg
     2221 ctcattccga gagcaacgtt caagttcaat aatagattcg ggttttgatt cctgtttgtg
     2281 ccgctcagtt tgatcgggat tgccgctcat ctcgtgtggt tttatgtgct tttgcaactg
     2341 gttagccacc tgttcgcgca gcgtttcgaa tgccactacc aaaggggtga gatcagcgat
     2401 atcgggcgtc ctcggtatcg cctggacgag gcttcggaaa tcttcctgca tagacgtcca
     2461 gtggcccggc acttctacga gcaccacgtc tatcaacttg gcgatgtctc tgcggcagag
     2521 cgtcagtcgc tcgcgtagcc gctgcagctc catccgatca gcctcggctt cggcagcgag
     2581 tcgctctatt tcgccggagc gggccagcag tggtgcgagc gaaaagccga acgctgtgcc
     2641 gatcgctcca tcgcgagtct tgcgtgcata gcgcttgccg ttcggactgt ccttgcgagc
     2701 gaccagccct gcatcgacga gggcggccag atgccttctg agcgtagcct cggccatgcc
     2761 atgagctctc agtgacagct gaacgttcga gggaaagacg accagactgg tctccccgct
     2821 tagcgtgttc tccggataga agctcaacag cgcgttcaga acggctagtg tccgatcgct
     2881 gatgccaaga cggcgtttgg cagcacaaag cgagcgaaac aatttccact tgtcgaccga
     2941 cttgccaggt tcgatgtcgc gggagatgaa ctggcctgcc aacagggcaa gcgtcatcga
     3001 ccgccgccca aagggcgtcg tcacacttcc aatctccatt tccttcacct tcgataaggc
     3061 aaaagaaagc agctcaccaa aaacggtgcc aaggactctt gactgtgatt cgtggaaatg
     3121 cgattctcga tcttgctaca aatctgagga aggcttccac ggcttgttcg ttgggggcct
     3181 ttttcttttg cgggttagtc tccttgtttc gaactctgct tttgcatgaa ctggtcgtgc
     3241 aggctttcga tctgacctgc aatgaaatcg acaaagcctg acggcacaga cttgttgaat
     3301 gtcagcttcg cagccgactt gccgcgttgg atctggccaa gcacgaggcc atgagagatc
     3361 agatcgctcg tggaagacgc tgcagccctc acatcaacct tggtaagggc ttcaaaaacc
     3421 gcagtgaagc gtagatcgct ggaagaggct tgaatctgct cttgctccag aaccaactgc
     3481 gccaccttcg cgggctcccg gtgagccgcc cagcgcactt cgaactcgat ccatctacgc
     3541 cgtccgattt caggagccgg accaatctgc atcaggatct cgcgcggcaa tgctgccgct
     3601 atccggatca tttcggagat atgcgacctg ccggtgctca gggacttgca gatcacgtcg
     3661 cggcgatagc cggcagcctc ctgcgattgc gcaaaaaggc atcgctcgat gaaggacagg
     3721 tccttgcggt tgctgttctc ttccccttgg agaacgatcg catcttcgtc gctaagttcc
     3781 cgggcgattg ccttgaactt gatgccgagg agtttggcag cggcaagccg gcgccggcca
     3841 aatacgatct gaaacggcct tgcggcccct cgaaccgggc ggaccaagcc aggagtactc
     3901 tgtccatgct cgcgcatcga ctcgagcaaa tccgcaatac cggcttcgct gtagccggaa
     3961 tcaaagcgat cccggatggc cgattccatg atctcgtctg gatcgatctc gacgatctgg
     4021 ccaccatcct tgagaaggcg ttcggcaagt tcgctgcgtt cctgcatctg acggacgccg
     4081 gctgcaactt tgagaagatg tggcgatgtc gaccgctgca tcccgccctt ctctgcagaa
     4141 ttttccagct gtccgaggac attggcaaaa aggcccttgg aatccttgcg gctcattctc
     4201 tcccccatgc cttccagaag aggccctcga gttcagcatt ggcagcattg atcgactcga
     4261 tggcgcggtc gtaggttttc ggcgccgaaa atcgggacct ctgaacctcg taaaggctct
     4321 gcttccacgt cagagcatcg gccaccgcgg tcgacttgat gacggagttg aggagcagat
     4381 cctcgccgaa cacctgtctc atcagcgcgg acatattcgc ttgcggatga tcgttcggct
     4441 cgaactttgt gatcaggaac ttggcgaagt cccattgagc ggcggtgccc acctcattca
     4501 ggatctgcat gtacgacccg gcaagctcca aaaactttgc ggtcgagtcc acatccagca
     4561 tatgtgccgg aaccgggatg atgactccag tcgcggcggt cagcgacgaa agtgtcagga
     4621 agttcagcga cggagccgag tccataagga cgaagtcgta gtcatcggcg acctgctcca
     4681 gagcttgaga aacgcgaagc aagaaaccgg ttcctgctga ccgccgcatc tcgagggcga
     4741 cggccgtctc gaactcggtc agttcgaggc cggcacagat gacgtcgaaa ccaacgatgt
     4801 gggtcttgtg gatgatggat ttcgtcggga cgggatcgtc gaaacgaatg gctgagaaaa
     4861 gtgtatcgcc gccgcggtaa tcgaattcgg gcaggctacc atggagcgcg gtgagagatg
     4921 cttgcgggtc gagatcgacc gccagcacgc gatagccctt gagtgccaga tagtgaccga
     4981 gatggattgt ggtagaggtc ttgctggagc cgcccttgaa gttggtgacc gcaacgatct
     5041 ggcattgctc attgcctgtc cggcgggggt tcatccattt cttccggccg tttgccgcaa
     5101 gagccatacg tagctcaaga acctgctcga gcgtgtagaa gcgccggccg ttggcgattt
     5161 cgggctcagg gccctgttcc ttggcggcaa cctggcggat ataggcctcg gttacgccca
     5221 gcagggtggc aacttcggta gaggtgaact tgcgcattgt gcgacgcgcg ttcgggggga
     5281 actgcgccat gctcaagttg ttcagtgcca tttcgagctt actggaaaaa ttctccatga
     5341 aagcgaggct gcttaggtgt cgcgtcatcg aaagaatccc tgtgatcgtt tccgtcactt
     5401 aagcagtgaa gtggttaacc gcttgctaat cgccacgtaa ctactcgaaa acacgcaaat
     5461 tcgaaaaatt tccgcaacta agacgtactc cgattcggct atggccatca atgcgcgctg
     5521 tggatgacgc acagagccgg ggggaaggcg ccagtgcatg ccggttctcg accctccccg
     5581 cctctttttt ggacgaagat gtgcgcagac gtcgctgggc ggagacgcaa tagtggttca
     5641 ctgcggaaat ggttggccgt tgctttcaat ggagatctgc cgaactcttc gcagagttcc
     5701 ccgcggggaa ctctcgaagc gacgacgctg gcgcgcagta ccgactgctc gccaaggcga
     5761 gacatatgtg ccgtattggg aacagctcgg catctcggtg ctcgcctgcg ctccggcctg
     5821 cctcaatctc gttggccatg accgagcaac gactgcaccg gatggttcag ttacgctagg
     5881 gataacaggg taatatagtc gagctggttg ccctcgccgc tgggctggcg gccgtctatg
     5941 gccctgcaaa cgcgccagaa acgccgtcga agccgtgtgc gagacaccgc ggccggccgc
     6001 cggcgttgtg gatacctcgc ggaaaacttg gccctcactg acagatgagg ggcggacgtt
     6061 gacacttgag gggccgactc acccggcgcg gcgttgacag atgaggggca ggctcgattt
     6121 cggccggcga cgtggagctg gccagcctcg caaatcggcg aaaacgcctg attttacgcg
     6181 agtttcccac agatgatgtg gacaagcctg gggataagtg ccctgcggta ttgacacttg
     6241 aggggcgcga ctactgacag atgaggggcg cgatccttga cacttgaggg gcagagtgct
     6301 gacagatgag gggcgcacct attgacattt gaggggctgt ccacaggcag aaaatccagc
     6361 atttgcaagg gtttccgccc gtttttcggc caccgctaac ctgtctttta acctgctttt
     6421 aaaccaatat ttataaacct tgtttttaac cagggctgcg ccctgtgcgc gtgaccgcgc
     6481 acgccgaagg ggggtgcccc cccttctcga accctcccgg tcgagtgagc gaggaagcac
     6541 cagggaacag cacttatata ttctgcttac acacgatgcc tgaaaaaact tcccttgggg
     6601 ttatccactt atccacgggg atatttttat aattattttt tttatagttt ttagatcttc
     6661 ttttttagag cgccttgtag gcctttatcc atgctggttc tagagaaggt gttgtgacaa
     6721 attgcccttt cagtgtgaca aatcaccctc aaatgacagt cctgtctgtg acaaattgcc
     6781 cttaaccctg tgacaaattg ccctcagaag aagctgtttt ttcacaaagt tatccctgct
     6841 tattgactct tttttattta gtgtgacaat ctaaaaactt gtcacacttc acatggatct
     6901 gtcatggcgg aaacagcggt tatcaatcac aagaaacgta aaaatagccc gcgaatcgtc
     6961 cagtcaaacg acctcactga ggcggcatat agtctctccc gggatcaaaa acgtatgctg
     7021 tatctgttcg ttgaccagat cagaaaatct gatggcaccc tacaggaaca tgacggtatc
     7081 tgcgagatcc atgttgctaa atatgctgaa atattcggat tgacctctgc ggaagccagt
     7141 aaggatatac ggcaggcatt gaagagtttc gcggggaagg aagtggtttt ttatcgccct
     7201 gaagaggatg ccggcgatga aaaaggctat gaatcttttc cttggtttat caaacgtgcg
     7261 cacagtccat ccagagggct ttacagtgta catatcaacc catatctcat tcccttcttt
     7321 atcgggttac agaaccggtt tacgcagttt cggcttagtg aaacaaaaga aatcaccaat
     7381 ccgtatgcca tgcgtttata cgaatccctg tgtcagtatc gtaagccgga tggctcaggc
     7441 atcgtctctc tgaaaatcga ctggatcata gagcgttacc agctgcctca aagttaccag
     7501 cgtatgcctg acttccgccg ccgcttcctg caggtctgtg ttaatgagat caacagcaga
     7561 actccaatgc gcctctcata cattgagaaa aagaaaggcc gccagacgac tcatatcgta
     7621 ttttccttcc gcgatatcac ttccatgacg acaggatagt ctgagggtta tctgtcacag
     7681 atttgagggt ggttcgtcac atttgttctg acctactgag ggtaatttgt cacagttttg
     7741 ctgtttcctt cagcctgcat ggattttctc atactttttg aactgtaatt tttaaggaag
     7801 ccaaatttga gggcagtttg tcacagttga tttccttctc tttcccttcg tcatgtgacc
     7861 tgatatcggg ggttagttcg tcatcattga tgagggttga ttatcacagt ttattactct
     7921 gaattggcta tccgcgtgtg tacctctacc tggagttttt cccacggtgg atatttcttc
     7981 ttgcgctgag cgtaagagct atctgacaga acagttcttc tttgcttcct cgccagttcg
     8041 ctcgctatgc tcggttacac ggctgcggcg agcatcacgt gctataaaaa taattataat
     8101 ttaaattttt taatataaat atataaatta aaaatagaaa gtaaaaaaag aaattaaaga
     8161 aaaaatagtt tttgttttcc gaagatgtaa aagactctag ggggatcgcc aacaaatact
     8221 accttttacc ttgctcttcc tgctctcagg tattaatgcc gaattgtttc atcttgtctg
     8281 tgtagaagac cacacacgaa aatcctgtga ttttacattt tacttatcgt taatcgaatg
     8341 tatatctatt taatctgctt ttcttgtcta ataaatatat atgtaaagta cgctttttgt
     8401 tgaaattttt taaacctttg tttatttttt tttcttcatt ccgtaactct tctaccttct
     8461 ttatttactt tctaaaatcc aaatacaaaa cataaaaata aataaacaca gagtaaattc
     8521 ccaaattatt ccatcattaa aagatacgag gcgcgtgtaa gttacaggca agcgatccta
     8581 gtacactcta tattttttta tgcctcggta atgattttca tttttttttt ccacctagcg
     8641 gatgactctt tttttttctt agcgattggc attatcacat aatgaattat acattatata
     8701 aagtaatgtg atttcttcga agaatatact aaaaaatgag caggcaagat aaacgaaggc
     8761 aaagatgaca gagcagaaag ccctagtaaa gcgtattaca aatgaaacca agattcagat
     8821 tgcgatctct ttaaagggtg gtcccctagc gatagagcac tcgatcttcc cagaaaaaga
     8881 ggcagaagca gtagcagaac aggccacaca atcgcaagtg attaacgtcc acacaggtat
     8941 agggtttctg gaccatatga tacatgctct ggccaagcat tccggctggt cgctaatcgt
     9001 tgagtgcatt ggtgacttac acatagacga ccatcacacc actgaagact gcgggattgc
     9061 tctcggtcaa gcttttaaag aggccctagg ggccgtgcgt ggagtaaaaa ggtttggatc
     9121 aggatttgcg cctttggatg aggcactttc cagagcggtg gtagatcttt cgaacaggcc
     9181 gtacgcagtt gtcgaacttg gtttgcaaag ggagaaagta ggagatctct cttgcgagat
     9241 gatcccgcat tttcttgaaa gctttgcaga ggctagcaga attaccctcc acgttgattg
     9301 tctgcgaggc aagaatgatc atcaccgtag tgagagtgcg ttcaaggctc ttgcggttgc
     9361 cataagagaa gccacctcgc ccaatggtac caacgatgtt ccctccacca aaggtgttct
     9421 tatgtagttt tacacaggag tctggacttg acgctagtga taataagtga ctgaggtatg
     9481 tgctcttctt atctcctttt gtagtgttgc tcttatttta aacaactttg cggttttttg
     9541 atgactttgc gattttgttg ttgctttgca gtaaattgca agatttaata aaaaaacgca
     9601 aagcaatgat taaaggatgt tcagaatgaa actcatggaa acacttaacc agtgcataaa
     9661 cgctggtcat gaaatgacga aggctatcgc cattgcacag tttaatgatg acagcccgga
     9721 agcgaggaaa ataacccggc gctggagaat aggtgaagca gcggatttag ttggggtttc
     9781 ttctcaggct atcagagatg ccgagaaagc agggcgacta ccgcacccgg atatggaaat
     9841 tcgaggacgg gttgagcaac gtgttggtta tacaattgaa caaattaatc atatgcgtga
     9901 tgtgtttggt acgcgattgc gacgtgctga agacgtattt ccaccggtga tcggggttgc
     9961 tgcccataaa ggtggcgttt acaaaacctc agtttctgtt catcttgctc aggatctggc
    10021 tctgaagggg ctacgtgttt tgctcgtgga aggtaacgac ccccagggaa cagcctcaat
    10081 gtatcacgga tgggtaccag atcttcatat tcatgcagaa gacactctcc tgcctttcta
    10141 tcttggggaa aaggacgatg tcacttatgc aataaagccc acttgctggc cggggcttga
    10201 cattattcct tcctgtctgg ctctgcaccg tattgaaact gagttaatgg gcaaatttga
    10261 tgaaggtaaa ctgcccaccg atccacacct gatgctccga ctggccattg aaactgttgc
    10321 tcatgactat gatgtcatag ttattgacag cgcgcctaac ctgggtatcg gcacgattaa
    10381 tgtcgtatgt gctgctgatg tgctgattgt tcccacgcct gctgagttgt ttgactacac
    10441 ctccgcactg cagtttttcg atatgcttcg tgatctgctc aagaacgttg atcttaaagg
    10501 gttcgagcct gatgtacgta ttttgcttac caaatacagc aatagcaatg gctctcagtc
    10561 cccgtggatg gaggagcaaa ttcgggatgc ctggggaagc atggttctaa aaaatgttgt
    10621 acgtgaaacg gatgaagttg gtaaaggtca gatccggatg agaactgttt ttgaacaggc
    10681 cattgatcaa cgctcttcaa ctggtgcctg gagaaatgct ctttctattt gggaacctgt
    10741 ctgcaatgaa attttcgatc gtctgattaa accacgctgg gagattagat aatgaagcgt
    10801 gcgcctgtta ttccaaaaca tacgctcaat actcaaccgg ttgaagatac ttcgttatcg
    10861 acaccagctg ccccgatggt ggattcgtta attgcgcgcg taggagtaat ggctcgcggt
    10921 aatgccatta ctttgcctgt atgtggtcgg gatgtgaagt ttactcttga agtgctccgg
    10981 ggtgatagtg ttgagaagac ctctcgggta tggtcaggta atgaacgtga ccaggagctg
    11041 cttactgagg acgcactgga tgatctcatc ccttcttttc tactgactgg tcaacagaca
    11101 ccggcgttcg gtcgaagagt atctggtgtc atagaaattg ccgatgggag tcgccgtcgt
    11161 aaagctgctg cacttaccga aagtgattat cgtgttctgg ttggcgagct ggatgatgag
    11221 cagatggctg cattatccag attgggtaac gattatcgcc caacaagtgc ttatgaacgt
    11281 ggtcagcgtt atgcaagccg attgcagaat gaatttgctg gaaatatttc tgcgctggct
    11341 gatgcggaaa atatttcacg taagattatt acccgctgta tcaacaccgc caaattgcct
    11401 aaatcagttg ttgctctttt ttctcacccc ggtgaactat ctgcccggtc aggtgatgca
    11461 cttcaaaaag cctttacaga taaagaggaa ttacttaagc agcaggcatc taaccttcat
    11521 gagcagaaaa aagctggggt gatatttgaa gctgaagaag ttatcactct tttaacttct
    11581 gtgcttaaaa cgtcatctgc atcaagaact agtttaagct cacgacatca gtttgctcct
    11641 ggagcgacag tattgtataa gggcgataaa atggtgctta acctggacag gtctcgtgtt
    11701 ccaactgagt gtatagagaa aattgaggcc attcttaagg aacttgaaaa gccagcaccc
    11761 tgatgcgacc tcgttttagt ctacgtttat ctgtctttac ttaatgtcct ttgttacagg
    11821 ccagaaagca taactggcct gaatattctc tctgggccca ctgttccact tgtatcgtcg
    11881 gtctgataat cagactggga ccacggtccc actcgtatcg tcggtctgat tattagtctg
    11941 ggaccacggt cccactcgta tcgtcggtct gattattagt ctgggaccac ggtcccactc
    12001 gtatcgtcgg tctgataatc agactgggac cacggtccca ctcgtatcgt cggtctgatt
    12061 attagtctgg gaccatggtc ccactcgtat cgtcggtctg attattagtc tgggaccacg
    12121 gtcccactcg tatcgtcggt ctgattatta gtctggaacc acggtcccac tcgtatcgtc
    12181 ggtctgatta ttagtctggg accacggtcc cactcgtatc gtcggtctga ttattagtct
    12241 gggaccacga tcccactcgt gttgtcggtc tgattatcgg tctgggacca cggtcccact
    12301 tgtattgtcg atcagactat cagcgtgaga ctacgattcc atcaatgcct gtcaagggca
    12361 agtattgaca tgtcgtcgta acctgtagaa cggagtaacc tcggtgtgcg gttgtatgcc
    12421 tgctgtggat tgctgctgtg tcctgcttat ccacaacatt ttgcgcacgg ttatgtggac
    12481 aaaatacctg gttacccagg ccgtgccggc acgttaaccg ggctgcatcc gatgcaagtg
    12541 tgtcgctgtc gacgagctcg cgagctcgga catgaggttg ccccgtattc agtgtcgctg
    12601 atttgtattg tctgaagttg tttttacgtt aagttgatgc agatcaatta atacgatacc
    12661 tgcgtcataa ttgattattt gacgtggttt gatggcctcc acgcacgttg tgatatgtag
    12721 atgataatca ttatcacttt acgggtcctt tccggtgatc cgacaggtta cggggcggcg
    12781 acctcgcggg ttttcgctat ttatgaaaat tttccggttt aaggcgtttc cgttcttctt
    12841 cgtcataact taatgttttt atttaaaata ccctctgaaa agaaaggaaa cgacaggtgc
    12901 tgaaagcgag ctttttggcc tctgtcgttt cctttctctg tttttgtccg tggaatgaac
    12961 aatggaagtc cgagctcatc gctaataact tcgtatagca tacattatac gaagttatat
    13021 tcgatgcggc cgcaaggggt tcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
    13081 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccaca
    13141 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcagct gcgcaactgt
    13201 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
    13261 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
    13321 acggccagtg aattgtaata cgactcacta tagggcgaat tcgagctcgg tacccgggga
    13381 tcctctagag tcgacctgca ggcatgcaag cttgagtatt ctatagtctc acctaaatag
    13441 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
    13501 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
    13561 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
    13621 gctgcattaa tgaatcggcc aacgcgaacc ccttgcggcc gcccgggccg tcgaccaatt
    13681 ctcatgtttg acagcttatc atcgaatttc tgccattcat ccgcttatta tcacttattc
    13741 aggcgtagca accaggcgtt taagggcacc aataactgcc ttaaaaaaat tacgccccgc
    13801 cctgccactc atcgcagtac tgttgtaatt cattaagcat tctgccgaca tggaagccat
    13861 cacaaacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat
    13921 aatatttgcc catggtgaaa acgggggcga agaagttgtc catattggcc acgtttaaat
    13981 caaaactggt gaaactcacc cagggattgg ctgagacgaa aaacatattc tcaataaacc
    14041 ctttagggaa ataggccagg ttttcaccgt aacacgccac atcttgcgaa tatatgtgta
    14101 gaaactgccg gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt tcagtttgct
    14161 catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca ccgtctttca
    14221 ttgccatacg aaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg
    14281 gataaaactt gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa
    14341 cggtctggtt ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat
    14401 gccattggga tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct
    14461 tagctcctga aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat
    14521 ggtgaaagtt ggaacctctt acgtgccgat caacgtctca ttttcgccaa aagttggccc
    14581 agggcttccc ggtatcaaca gggacaccag gatttattta ttctgcgaag tgatcttccg
    14641 tcacaggtat ttattcgcga taagctcatg gagcggcgta accgtcgcac aggaaggaca
    14701 gagaaagcgc ggatctggga agtgacggac agaacggtca ggacctggat tggggaggcg
    14761 gttgccgccg ctgctgctga cggtgtgacg ttctctgttc cggtcacacc acatacgttc
    14821 cgccattcct atgcgatgca catgctgtat gccggtatac cgctgaaagt tctgcaaagc
    14881 ctgatgggac ataagtccat cagttcaacg gaagtctaca cgaaggtttt tgcgctggat
    14941 gtggctgccc ggcaccgggt gcagtttgcg atgccggagt ctgatgcggt tgcgatgctg
    15001 aaacaattat cctgagaata aatgccttgg cctttatatg gaaatgtgga actgagtgga
    15061 tatgctgttt ttgtctgtta aacagagaag ctggctgtta tccactgaga agcgaacgaa
    15121 acagtcggga aaatctccca ttatcgtaga gatccgcatt attaatctca ggagcctgtg
    15181 tagcgtttat aggaagtagt gttctgtcat gatgcctgca agcggtaacg aaaacgattt
    15241 gaatatgcct tcaggaacaa tagaaatctt cgtgcggtgt tacgttgaag tggagcggat
    15301 tatgtcagca atggacagaa caacctaatg aacacagaac catgatgtgg tctgtccttt
    15361 tacagccagt agtgctcgcc gcagtcgagc gacagggcga agcccgttta aacggggtgg
    15421 gcgaagaact ccagcatgag atccccgcgc tggaggatca tccagccggc gtcccggaaa
    15481 acgattccga agcccaacct ttcatagaag gcggcggtgg aatcgaaatc tcgtgatggc
    15541 aggttgggcg tcgcttggtc ggtcatttcg aaccccagag tcccgctcag aagaactcgt
    15601 caagaaggcg atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga
    15661 ggaagcggtc agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgcta
    15721 tgtcctgata gcggtccgcc acacccagcc ggccacagtc gatgaatcca gaaaagcggc
    15781 cattttccac catgatattc ggcaagcagg catcgccatg ggtcacgacg agatcctcgc
    15841 cgtcgggcat ccgcgccttg agcctggcga acagttcggc tggcgcgagc ccctgatgct
    15901 cttcgtccag atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga
    15961 tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc
    16021 gcattgcatc agccatgatg gatactttct cggcaggagc aaggtgagat gacaggagat
    16081 cctgccccgg cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga
    16141 gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtctt
    16201 ggagttcatt cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg
    16261 ctgacagccg gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc
    16321 cgaatagcct ctccacccaa gcggccggag aacctgcgtg caatccatct tgttcaatca
    16381 tgcgaaacga tcctcatcct gtctcttgat cagatcttga tcccctgcgc catcagatcc
    16441 ttggcggcaa gaaagccatc cagtttactt tgcagggctt cccaacctta ccagagggcg
    16501 ccccagctgg caattccggt tcgcttgctg tccataaaac cgcccagtct agctatcgcc
    16561 atgtaagccc actgcaagct acctgctttc tctttgcgct tgcgttttcc cttgtccaga
    16621 tagcccagta gctgacattc atccggggtc agcaccgttt ctgcggactg gctttctacg
    16681 tgttccgctt cctttagcag cccttgcgcc ctgagtgctt gcggcagcgt gaagctttaa
    16741 ttaaatcttc cgctgcataa ccctgcttcg gggtcattat agcgattttt tcggtatatc
    16801 catccttttt cgcacgatat acaggatttt gccaaagggt tcgtgtagac tttccttggt
    16861 gtatccaacg gcgtcagccg ggcaggatag gtgaagtagg cccacccgcg agcgggtgtt
    16921 ccttcttcac tgtcccttat tcgcacctgg cggtgctcaa cgggaatcct gctctgcgag
    16981 gctggccggc taccgccggc gtaacagatg agggcaagcg gatggctgat gaaaccaagc
    17041 caaccaggaa gggcagccca cctatcaagg tgtactgcct tccagacgaa cgaagagcga
    17101 ttgaggaaaa ggcggcggcg gccggcatga gcctgtcggc ctacctgctg gccgtcggcc
    17161 agggctacaa aatcacgggc gtcgtggact atgagcacgt ccgcgagctg gcccgcatca
    17221 atggcgacct gggccgcctg ggcggcctgc tgaaactctg gctcaccgac gacccgcgca
    17281 cggcgcggtt cggtgatgcc acgatcctcg ccctgctggc gaagatcgaa gagaagcagg
    17341 acgagcttgg caaggtcatg atgggcgtgg tccgcccgag ggcagagcca tgactttttt
    17401 agccgctaaa acggccgggg ggtgcgcgtg attgccaagc acgtccccat gcgctccatc
    17461 aagaagaggc acttcgagct gtaagtacat caccgacgag caaggcaaga cgatc
//
