LOCUS       sequence-375342-       18420 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 pSymB origin and Spec resistance..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 18420)
  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 18420)
  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: pBGE1.0
FEATURES             Location/Qualifiers
     source          1..18420
                     /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      6066..6680
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     rep_origin      6157..6688
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      6756..6975
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             7066..7821
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     misc_feature    7824..8074
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     misc_feature    8236..8739
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        8740..8926
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             8927..9589
                     /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             9779..10954
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     CDS             10954..11925
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     misc_feature    11998..12471
                     /gene="sopC"
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     misc_feature    12731..13129
                     /label=cos
                     /note="lambda cos site; allows packaging into phage lambda 
                     particles"
     protein_bind    complement(13147..13180)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     13248..13267
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(13347..13622)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSYLGETIEYSSLHACRSTLEDPRVPSSNSPYSESYYNSL
                     AVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRS"
     primer_bind     13460..13482
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     13474..13491
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     13475..13491
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     13498..13517
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        13498..13516
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    13519..13575
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(13618..13634)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13618..13634)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(13631..13653)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    13642..13658
                     /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(13666..13672,13673..13690,13691..13696))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13711..13732
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(13952..14611)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     14526..14545
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(14612..14714)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             16453..17244
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     oriT            17785..17894
                     /note="incP origin of transfer"
     CDS             17927..18298
                     /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 gaagccttat ccttctctta tccgacctgc
     1561 ggggctcgat cctgtcacaa tccggtccgc tgtcaatagt ctactaaaat tatgatctac
     1621 taagggtacc gcgtctgttg caacgtctgg gggagatttg cgcggccgct cggatgcggc
     1681 gcttcgccaa cgcaagtgtc ggccggatcg gcccggctgg actgccggcg agttgtccgc
     1741 ggacaaccaa tgaacagccg gacgacaaaa ctgcagcgcc cctgcagcac aaccgagctt
     1801 tttgttaagc gtttgtaaaa tggcaacgaa tcggcgcata gtataactgc gcagatcagc
     1861 tcccattggt gaattttgcg cagtttttgg agagtaggaa tgttgcagca gagaatccag
     1921 agtcttgagg agcaggagca tttgccgtcg ctcttgtctg ccgacgctca ggaactcgca
     1981 caccagttgc aggagcatca aaagaagatc ttttctccgt cggcgcaaaa gacgatgcgg
     2041 ctcttcagcc cggcagaggc agcggccttc atcggaatcg gcgaaggcta tctgcgccag
     2101 atcgcctcag aaggccatgg tccgcagccg ctgtcaaacg gccgccgcat gtatgcgatg
     2161 gaggacatcg accgtatccg ccgggttctc gacgaaggca gcaagaccgg caaatatatt
     2221 ccgcatcgtc gaccgggcga aaagctgcaa gtcgtctccg tcatgaattt caaaggtggt
     2281 tcgggcaaga cgacgacatc cgctcacctt gcgcagtatc tcgccttgcg cggctatcgg
     2341 gtgctggcga tcgatctcga tccgcaggcc tcgctctccg ccctcttcgg tcatcagccg
     2401 gaactggacg tcggcgaagg ggaaacactc tatggcgcca tacgctacga ggccccccgg
     2461 ccgatcgcgg aggtggtgcg atcgacctat accgccaatc ttcacctcat tccgggcaat
     2521 ctcgagctca tggaattcga gcacgagact ccgaaggcga tgatcgcggg ctccaccgag
     2581 accatgttct tcgcgcggat cggcgaggtg ctttccgaaa tcgagagctt ctacgacatc
     2641 gtcgtcatcg actgcccgcc gcagctcggc tttctgacca tgtcggcgct ttgcgccgcg
     2701 acctccgtgc tcatcaccgt gcacccgcag atgctcgacg tcatgtcgat gtcccagttc
     2761 ttgtcgatga cgagcgagct catggccgtt gtcgagaatg ccggcgggcg caccagctat
     2821 gactggatgc gctatctggt cacgcgcttc gagcccaacg acggaccgca gagccaaatg
     2881 accggtttca tgcgcgcgat cttcggcaac cgcatgctgc agaacgcgat ggtgaaatcg
     2941 accgcaattt ccgacgccgg cgtcaccaag cagacgctct atgaagtgga gcgctcgcaa
     3001 ttcatacgcg gtacctacga ccgggcgatg gattcgctct cgctcgtcaa tgcggagatc
     3061 gaagagatga tccgcagggt ctgggggagg aagtaaccga tggcgcgcaa gaacctgatc
     3121 ggcatttccg agagccccct cgcccccgaa gacggggagc gcccggccgt cggtcgtccg
     3181 atcgcgggcc ttgcgcccgc ccagcgctcc actggcctcg tcggcggcat caccagatcc
     3241 ctctcgaaca tcacccagaa ggtcgagcga gcggaggaat tggagcgcca gcttgccgat
     3301 gggcacgcaa tcgtcgaact ggacccggag ctgatcgatt cctctttcat catcgatagg
     3361 ctgggtgtcg cccccgacgt tcaggccatg ctggttcagc agattcggga ccacggacaa
     3421 caggttccga tcctcgtgcg gccgcatccg gcggagagcg gccgatatca ggtcgcctac
     3481 ggccacaggc ggcttgcggc attgcgcgag atcggcatca aggtcaaggc agtcatccgg
     3541 aacttgaacg acgagcaact cgtcatttcc caggggcagg aaaacaacgc ccgcaccgac
     3601 ctctccttca tagagcgagc gttgttcgca acgcgtcttg aagatcgcgg cttctcgcgg
     3661 gatacgatca tgtcggccct cggagtcgac aaggcggccc tctccaagat gatcgccgtc
     3721 gtccgccgat tgccgttgga catcatcgaa gccatcggtg ccgcgccggg tttcggcagg
     3781 agacgatgga tggaacttgc cgatctcatc gaccgcgatg accggcgtcc gaaggcattg
     3841 catttcatcg agacggccgc attcaaggcg ctggagagcg atcagcgttt cgaaaagctg
     3901 tttgccttgc tgagcctagc caggaagccc tcaaaggcgg aaacctggat tgctccggac
     3961 aggagccgcc ccgtgcgtat tcgcgaaacg gatgcggaaa cgacgctggc cttcagcaag
     4021 aaggcggcac cgggctttgc agaattcgtg aggcagaggc tcgaagcgct ctacctcgaa
     4081 tatcaacggg aaacaggaga ctaaacagag caaaagaaaa aggcttccga acgactagcg
     4141 ctgcggaaac ccttctctcg tgtagcaact agagaatccc acttccgcga atcagtgtca
     4201 agcatttttg acgtcgtttc ggcgaacgga tttcttttgc cttggaaaag gtgaagaaaa
     4261 tggaacgtgg aagtgtgacg acgccctttg ggcggcgggc gatgacgctt ggcatgctcg
     4321 caagtcagat cgccgccggt aacgtcccgc cggaccggtc ggtggacaag tggaaattgt
     4381 tccgttctct ctgtgaagcg aagacggtga tcggcgtgtc cgatcgttcg cttgcggtgt
     4441 tgaacgccct tttgagcttt tatccaggag ccgagctctc gggcgaaaat ggcctcgtcg
     4501 ttttcccctc caatgcgcag ctctcgctcc gtacccacgg aatggcgggc acgacgctcc
     4561 gccggcatct tgcatgcctc gtggaatcgg gcctgatcct gcggcgcgac agcccgaacg
     4621 gtaagcgtta cgcccgaagg agccgtaatg gagcggtggg cgaagccttc ggattcgacc
     4681 tctccccgct cgtcgcccgg gccgatgagt tcaaagccgc cgctgccgaa cttgccgcag
     4741 ccaaacagta tcatcgtctg atgcgcgaaa gactgagcct ctgccggcgc gacatcgcca
     4801 agctgctcga gctccttcgt gactctgacc tgcccaccga ctgcgagcgg cagcaacgga
     4861 gatatgagga attgtctgcg cctctctcgc gcaaggcacc ggtcagcgac gtcgagaccc
     4921 ttcatgacgc tatggcggaa cttcgtacgg atctatccaa tcaattggaa aaactgctta
     4981 attccaaaaa aatgggcgcc aatgatcgcc ataatgggca gcacatacag agttcacatc
     5041 cagactctca atttgaatct gaacctggct tgggaaaagg gtcggggaaa ttttcggccg
     5101 gcgcccaaag ctgcaacgcg gaccctgtga caccgacggc gaagctctcg gaggtttcga
     5161 cccgtatcga ttcgcagcag ccggtcagct cggtgtcccc atctctgccg atctcccgcc
     5221 tcgcccggga aattcctctg cccgtcctcc tatccgcttg cccgcagatc gccgattacg
     5281 gaccacaggg ccgtataacg aactggaagg atctgctgac cgctgcagtc gtcgtcagaa
     5341 ccatgctgaa cgtgagcacc gatgcctatg acgatgcatg cacggttctc ggccaggaaa
     5401 atgctgcgac cgtcatcgct tgcatcctgg agagagccga ttgcataagt tcgccaggcg
     5461 gctatcttcg ggttcttacc gcgagagcgc ggcgccaggc tttctcggtt gctccgatgg
     5521 tcgctgcact gaggcgcgcg cgaggggcaa gcatgccgct tgcgagttga tgtctccgct
     5581 gaacagtctt tgtaagacag gaatctcggg cgcactgcag gagttgtcgg tcgcaaggca
     5641 cgtcaggctt gcgacgctga ccgctcgcgg ttgccttgac gctccagata ggggaaactc
     5701 gcttcggctg cggattcgca cttgctttga attgccgcct tggtgtctgt ttcttaacaa
     5761 ttttgcttct atcgtaacgt attggcaaaa aaggcggtaa gccggcaatg cggaagttaa
     5821 atcgccatta agttccgtca cgcggacggc cccagttcga ggcttggtga ccgcaacaac
     5881 agttatcaac cgcctcccct attctgcaat tgagtggaaa tctcgtgcgg tcggccacgc
     5941 aatccggggc aaagccgccg acgatatgca gatgccgatc cctgcagtcg tttccgcagt
     6001 tgacagctgt caacaatacg atcttgcccc agttacgcta gggataacag ggtaatatag
     6061 tcgagctggt tgccctcgcc gctgggctgg cggccgtcta tggccctgca aacgcgccag
     6121 aaacgccgtc gaagccgtgt gcgagacacc gcggccggcc gccggcgttg tggatacctc
     6181 gcggaaaact tggccctcac tgacagatga ggggcggacg ttgacacttg aggggccgac
     6241 tcacccggcg cggcgttgac agatgagggg caggctcgat ttcggccggc gacgtggagc
     6301 tggccagcct cgcaaatcgg cgaaaacgcc tgattttacg cgagtttccc acagatgatg
     6361 tggacaagcc tggggataag tgccctgcgg tattgacact tgaggggcgc gactactgac
     6421 agatgagggg cgcgatcctt gacacttgag gggcagagtg ctgacagatg aggggcgcac
     6481 ctattgacat ttgaggggct gtccacaggc agaaaatcca gcatttgcaa gggtttccgc
     6541 ccgtttttcg gccaccgcta acctgtcttt taacctgctt ttaaaccaat atttataaac
     6601 cttgttttta accagggctg cgccctgtgc gcgtgaccgc gcacgccgaa ggggggtgcc
     6661 cccccttctc gaaccctccc ggtcgagtga gcgaggaagc accagggaac agcacttata
     6721 tattctgctt acacacgatg cctgaaaaaa cttcccttgg ggttatccac ttatccacgg
     6781 ggatattttt ataattattt tttttatagt ttttagatct tcttttttag agcgccttgt
     6841 aggcctttat ccatgctggt tctagagaag gtgttgtgac aaattgccct ttcagtgtga
     6901 caaatcaccc tcaaatgaca gtcctgtctg tgacaaattg cccttaaccc tgtgacaaat
     6961 tgccctcaga agaagctgtt ttttcacaaa gttatccctg cttattgact cttttttatt
     7021 tagtgtgaca atctaaaaac ttgtcacact tcacatggat ctgtcatggc ggaaacagcg
     7081 gttatcaatc acaagaaacg taaaaatagc ccgcgaatcg tccagtcaaa cgacctcact
     7141 gaggcggcat atagtctctc ccgggatcaa aaacgtatgc tgtatctgtt cgttgaccag
     7201 atcagaaaat ctgatggcac cctacaggaa catgacggta tctgcgagat ccatgttgct
     7261 aaatatgctg aaatattcgg attgacctct gcggaagcca gtaaggatat acggcaggca
     7321 ttgaagagtt tcgcggggaa ggaagtggtt ttttatcgcc ctgaagagga tgccggcgat
     7381 gaaaaaggct atgaatcttt tccttggttt atcaaacgtg cgcacagtcc atccagaggg
     7441 ctttacagtg tacatatcaa cccatatctc attcccttct ttatcgggtt acagaaccgg
     7501 tttacgcagt ttcggcttag tgaaacaaaa gaaatcacca atccgtatgc catgcgttta
     7561 tacgaatccc tgtgtcagta tcgtaagccg gatggctcag gcatcgtctc tctgaaaatc
     7621 gactggatca tagagcgtta ccagctgcct caaagttacc agcgtatgcc tgacttccgc
     7681 cgccgcttcc tgcaggtctg tgttaatgag atcaacagca gaactccaat gcgcctctca
     7741 tacattgaga aaaagaaagg ccgccagacg actcatatcg tattttcctt ccgcgatatc
     7801 acttccatga cgacaggata gtctgagggt tatctgtcac agatttgagg gtggttcgtc
     7861 acatttgttc tgacctactg agggtaattt gtcacagttt tgctgtttcc ttcagcctgc
     7921 atggattttc tcatactttt tgaactgtaa tttttaagga agccaaattt gagggcagtt
     7981 tgtcacagtt gatttccttc tctttccctt cgtcatgtga cctgatatcg ggggttagtt
     8041 cgtcatcatt gatgagggtt gattatcaca gtttattact ctgaattggc tatccgcgtg
     8101 tgtacctcta cctggagttt ttcccacggt ggatatttct tcttgcgctg agcgtaagag
     8161 ctatctgaca gaacagttct tctttgcttc ctcgccagtt cgctcgctat gctcggttac
     8221 acggctgcgg cgagcatcac gtgctataaa aataattata atttaaattt tttaatataa
     8281 atatataaat taaaaataga aagtaaaaaa agaaattaaa gaaaaaatag tttttgtttt
     8341 ccgaagatgt aaaagactct agggggatcg ccaacaaata ctacctttta ccttgctctt
     8401 cctgctctca ggtattaatg ccgaattgtt tcatcttgtc tgtgtagaag accacacacg
     8461 aaaatcctgt gattttacat tttacttatc gttaatcgaa tgtatatcta tttaatctgc
     8521 ttttcttgtc taataaatat atatgtaaag tacgcttttt gttgaaattt tttaaacctt
     8581 tgtttatttt tttttcttca ttccgtaact cttctacctt ctttatttac tttctaaaat
     8641 ccaaatacaa aacataaaaa taaataaaca cagagtaaat tcccaaatta ttccatcatt
     8701 aaaagatacg aggcgcgtgt aagttacagg caagcgatcc tagtacactc tatatttttt
     8761 tatgcctcgg taatgatttt catttttttt ttccacctag cggatgactc tttttttttc
     8821 ttagcgattg gcattatcac ataatgaatt atacattata taaagtaatg tgatttcttc
     8881 gaagaatata ctaaaaaatg agcaggcaag ataaacgaag gcaaagatga cagagcagaa
     8941 agccctagta aagcgtatta caaatgaaac caagattcag attgcgatct ctttaaaggg
     9001 tggtccccta gcgatagagc actcgatctt cccagaaaaa gaggcagaag cagtagcaga
     9061 acaggccaca caatcgcaag tgattaacgt ccacacaggt atagggtttc tggaccatat
     9121 gatacatgct ctggccaagc attccggctg gtcgctaatc gttgagtgca ttggtgactt
     9181 acacatagac gaccatcaca ccactgaaga ctgcgggatt gctctcggtc aagcttttaa
     9241 agaggcccta ggggccgtgc gtggagtaaa aaggtttgga tcaggatttg cgcctttgga
     9301 tgaggcactt tccagagcgg tggtagatct ttcgaacagg ccgtacgcag ttgtcgaact
     9361 tggtttgcaa agggagaaag taggagatct ctcttgcgag atgatcccgc attttcttga
     9421 aagctttgca gaggctagca gaattaccct ccacgttgat tgtctgcgag gcaagaatga
     9481 tcatcaccgt agtgagagtg cgttcaaggc tcttgcggtt gccataagag aagccacctc
     9541 gcccaatggt accaacgatg ttccctccac caaaggtgtt cttatgtagt tttacacagg
     9601 agtctggact tgacgctagt gataataagt gactgaggta tgtgctcttc ttatctcctt
     9661 ttgtagtgtt gctcttattt taaacaactt tgcggttttt tgatgacttt gcgattttgt
     9721 tgttgctttg cagtaaattg caagatttaa taaaaaaacg caaagcaatg attaaaggat
     9781 gttcagaatg aaactcatgg aaacacttaa ccagtgcata aacgctggtc atgaaatgac
     9841 gaaggctatc gccattgcac agtttaatga tgacagcccg gaagcgagga aaataacccg
     9901 gcgctggaga ataggtgaag cagcggattt agttggggtt tcttctcagg ctatcagaga
     9961 tgccgagaaa gcagggcgac taccgcaccc ggatatggaa attcgaggac gggttgagca
    10021 acgtgttggt tatacaattg aacaaattaa tcatatgcgt gatgtgtttg gtacgcgatt
    10081 gcgacgtgct gaagacgtat ttccaccggt gatcggggtt gctgcccata aaggtggcgt
    10141 ttacaaaacc tcagtttctg ttcatcttgc tcaggatctg gctctgaagg ggctacgtgt
    10201 tttgctcgtg gaaggtaacg acccccaggg aacagcctca atgtatcacg gatgggtacc
    10261 agatcttcat attcatgcag aagacactct cctgcctttc tatcttgggg aaaaggacga
    10321 tgtcacttat gcaataaagc ccacttgctg gccggggctt gacattattc cttcctgtct
    10381 ggctctgcac cgtattgaaa ctgagttaat gggcaaattt gatgaaggta aactgcccac
    10441 cgatccacac ctgatgctcc gactggccat tgaaactgtt gctcatgact atgatgtcat
    10501 agttattgac agcgcgccta acctgggtat cggcacgatt aatgtcgtat gtgctgctga
    10561 tgtgctgatt gttcccacgc ctgctgagtt gtttgactac acctccgcac tgcagttttt
    10621 cgatatgctt cgtgatctgc tcaagaacgt tgatcttaaa gggttcgagc ctgatgtacg
    10681 tattttgctt accaaataca gcaatagcaa tggctctcag tccccgtgga tggaggagca
    10741 aattcgggat gcctggggaa gcatggttct aaaaaatgtt gtacgtgaaa cggatgaagt
    10801 tggtaaaggt cagatccgga tgagaactgt ttttgaacag gccattgatc aacgctcttc
    10861 aactggtgcc tggagaaatg ctctttctat ttgggaacct gtctgcaatg aaattttcga
    10921 tcgtctgatt aaaccacgct gggagattag ataatgaagc gtgcgcctgt tattccaaaa
    10981 catacgctca atactcaacc ggttgaagat acttcgttat cgacaccagc tgccccgatg
    11041 gtggattcgt taattgcgcg cgtaggagta atggctcgcg gtaatgccat tactttgcct
    11101 gtatgtggtc gggatgtgaa gtttactctt gaagtgctcc ggggtgatag tgttgagaag
    11161 acctctcggg tatggtcagg taatgaacgt gaccaggagc tgcttactga ggacgcactg
    11221 gatgatctca tcccttcttt tctactgact ggtcaacaga caccggcgtt cggtcgaaga
    11281 gtatctggtg tcatagaaat tgccgatggg agtcgccgtc gtaaagctgc tgcacttacc
    11341 gaaagtgatt atcgtgttct ggttggcgag ctggatgatg agcagatggc tgcattatcc
    11401 agattgggta acgattatcg cccaacaagt gcttatgaac gtggtcagcg ttatgcaagc
    11461 cgattgcaga atgaatttgc tggaaatatt tctgcgctgg ctgatgcgga aaatatttca
    11521 cgtaagatta ttacccgctg tatcaacacc gccaaattgc ctaaatcagt tgttgctctt
    11581 ttttctcacc ccggtgaact atctgcccgg tcaggtgatg cacttcaaaa agcctttaca
    11641 gataaagagg aattacttaa gcagcaggca tctaaccttc atgagcagaa aaaagctggg
    11701 gtgatatttg aagctgaaga agttatcact cttttaactt ctgtgcttaa aacgtcatct
    11761 gcatcaagaa ctagtttaag ctcacgacat cagtttgctc ctggagcgac agtattgtat
    11821 aagggcgata aaatggtgct taacctggac aggtctcgtg ttccaactga gtgtatagag
    11881 aaaattgagg ccattcttaa ggaacttgaa aagccagcac cctgatgcga cctcgtttta
    11941 gtctacgttt atctgtcttt acttaatgtc ctttgttaca ggccagaaag cataactggc
    12001 ctgaatattc tctctgggcc cactgttcca cttgtatcgt cggtctgata atcagactgg
    12061 gaccacggtc ccactcgtat cgtcggtctg attattagtc tgggaccacg gtcccactcg
    12121 tatcgtcggt ctgattatta gtctgggacc acggtcccac tcgtatcgtc ggtctgataa
    12181 tcagactggg accacggtcc cactcgtatc gtcggtctga ttattagtct gggaccatgg
    12241 tcccactcgt atcgtcggtc tgattattag tctgggacca cggtcccact cgtatcgtcg
    12301 gtctgattat tagtctggaa ccacggtccc actcgtatcg tcggtctgat tattagtctg
    12361 ggaccacggt cccactcgta tcgtcggtct gattattagt ctgggaccac gatcccactc
    12421 gtgttgtcgg tctgattatc ggtctgggac cacggtccca cttgtattgt cgatcagact
    12481 atcagcgtga gactacgatt ccatcaatgc ctgtcaaggg caagtattga catgtcgtcg
    12541 taacctgtag aacggagtaa cctcggtgtg cggttgtatg cctgctgtgg attgctgctg
    12601 tgtcctgctt atccacaaca ttttgcgcac ggttatgtgg acaaaatacc tggttaccca
    12661 ggccgtgccg gcacgttaac cgggctgcat ccgatgcaag tgtgtcgctg tcgacgagct
    12721 cgcgagctcg gacatgaggt tgccccgtat tcagtgtcgc tgatttgtat tgtctgaagt
    12781 tgtttttacg ttaagttgat gcagatcaat taatacgata cctgcgtcat aattgattat
    12841 ttgacgtggt ttgatggcct ccacgcacgt tgtgatatgt agatgataat cattatcact
    12901 ttacgggtcc tttccggtga tccgacaggt tacggggcgg cgacctcgcg ggttttcgct
    12961 atttatgaaa attttccggt ttaaggcgtt tccgttcttc ttcgtcataa cttaatgttt
    13021 ttatttaaaa taccctctga aaagaaagga aacgacaggt gctgaaagcg agctttttgg
    13081 cctctgtcgt ttcctttctc tgtttttgtc cgtggaatga acaatggaag tccgagctca
    13141 tcgctaataa cttcgtatag catacattat acgaagttat attcgatgcg gccgcaaggg
    13201 gttcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
    13261 gattgtactg agagtgcacc atatgcggtg tgaaatacca cacagatgcg taaggagaaa
    13321 ataccgcatc aggcgccatt cgccattcag ctgcgcaact gttgggaagg gcgatcggtg
    13381 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
    13441 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattgtaa
    13501 tacgactcac tatagggcga attcgagctc ggtacccggg gatcctctag agtcgacctg
    13561 caggcatgca agcttgagta ttctatagtc tcacctaaat agcttggcgt aatcatggtc
    13621 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
    13681 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
    13741 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
    13801 ccaacgcgaa ccccttgcgg ccgcccgggc cgtcgaccaa ttctcatgtt tgacagctta
    13861 tcatcgaatt tctgccattc atccgcttat tatcacttat tcaggcgtag caaccaggcg
    13921 tttaagggca ccaataactg ccttaaaaaa attacgcccc gccctgccac tcatcgcagt
    13981 actgttgtaa ttcattaagc attctgccga catggaagcc atcacaaacg gcatgatgaa
    14041 cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt ataatatttg cccatggtga
    14101 aaacgggggc gaagaagttg tccatattgg ccacgtttaa atcaaaactg gtgaaactca
    14161 cccagggatt ggctgagacg aaaaacatat tctcaataaa ccctttaggg aaataggcca
    14221 ggttttcacc gtaacacgcc acatcttgcg aatatatgtg tagaaactgc cggaaatcgt
    14281 cgtggtattc actccagagc gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac
    14341 aagggtgaac actatcccat atcaccagct caccgtcttt cattgccata cgaaattccg
    14401 gatgagcatt catcaggcgg gcaagaatgt gaataaaggc cggataaaac ttgtgcttat
    14461 ttttctttac ggtctttaaa aaggccgtaa tatccagctg aacggtctgg ttataggtac
    14521 attgagcaac tgactgaaat gcctcaaaat gttctttacg atgccattgg gatatatcaa
    14581 cggtggtata tccagtgatt tttttctcca ttttagcttc cttagctcct gaaaatctcg
    14641 ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc
    14701 ttacgtgccg atcaacgtct cattttcgcc aaaagttggc ccagggcttc ccggtatcaa
    14761 cagggacacc aggatttatt tattctgcga agtgatcttc cgtcacaggt atttattcgc
    14821 gataagctca tggagcggcg taaccgtcgc acaggaagga cagagaaagc gcggatctgg
    14881 gaagtgacgg acagaacggt caggacctgg attggggagg cggttgccgc cgctgctgct
    14941 gacggtgtga cgttctctgt tccggtcaca ccacatacgt tccgccattc ctatgcgatg
    15001 cacatgctgt atgccggtat accgctgaaa gttctgcaaa gcctgatggg acataagtcc
    15061 atcagttcaa cggaagtcta cacgaaggtt tttgcgctgg atgtggctgc ccggcaccgg
    15121 gtgcagtttg cgatgccgga gtctgatgcg gttgcgatgc tgaaacaatt atcctgagaa
    15181 taaatgcctt ggcctttata tggaaatgtg gaactgagtg gatatgctgt ttttgtctgt
    15241 taaacagaga agctggctgt tatccactga gaagcgaacg aaacagtcgg gaaaatctcc
    15301 cattatcgta gagatccgca ttattaatct caggagcctg tgtagcgttt ataggaagta
    15361 gtgttctgtc atgatgcctg caagcggtaa cgaaaacgat ttgaatatgc cttcaggaac
    15421 aatagaaatc ttcgtgcggt gttacgttga agtggagcgg attatgtcag caatggacag
    15481 aacaacctaa tgaacacaga accatgatgt ggtctgtcct tttacagcca gtagtgctcg
    15541 ccgcagtcga gcgacagggc gaagcccgtt taaacccggg gatccggtga ttgattgagc
    15601 aagctttatg cttgtaaacc gttttgtgaa aaaattttta aaataaaaaa ggggacctct
    15661 agggtcccca attaattagt aatataatct attaaaggtc attcaaaagg tcatccaccg
    15721 gatcaattcc cctgctcgcg caggctgggt gccaagctct cgggtaacat caaggcccga
    15781 tccttggagc ccttgccctc ccgcacgatg atcgtgccgt gatcgaaatc cagatccttg
    15841 acccgcagtt gcaaaccctc actgatccgc atgcccgttc catacagaag ctgggcgaac
    15901 aaacgatgct cgccttccag aaaaccgagg atgcgaacca cttcatccgg ggtcagcacc
    15961 accggcaagc gccgcgacgg ccgaggtctt ccgatctcct gaagccaggg cagatccgtg
    16021 cacagcacct tgccgtagaa gaacagcaag gccgccaatg cctgacgatg cgtggagacc
    16081 gaaaccttgc gctcgttcgc cagccaggac agaaatgcct cgacttcgct gctgcccaag
    16141 gttgccgggt gacgcacacc gtggaaacgg atgaaggcac gaacccagtg gacataagcc
    16201 tgttcggttc gtaagctgta atgcaagtag cgtatgcgct cacgcaactg gtccagaacc
    16261 ttgaccgaac gcagcggtgg taacggcgca gtggcggttt tcatggcttg ttatgactgt
    16321 ttttttgggg tacagtctat gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt
    16381 cgatgtttga tgttatggag cagcaacgat gttacgcagc agggcagtcg ccctaaaaca
    16441 aagttaaaca tcatgaggga agcggtgatc gccgaagtat cgactcaact atcagaggta
    16501 gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt gtacggctcc
    16561 gcagtggatg gcggcctgaa gccacacagt gatattgatt tgctggttac ggtgaccgta
    16621 aggcttgatg aaacaacgcg gcgagctttg atcaacgacc ttttggaaac ttcggcttcc
    16681 cctggagaga gcgagattct ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc
    16741 attccgtggc gttatccagc taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac
    16801 attcttgcag gtatcttcga gccagccacg atcgacattg atctggctat cttgctgaca
    16861 aaagcaagag aacatagcgt tgccttggta ggtccagcgg cggaggaact ctttgatccg
    16921 gttcctgaac aggatctatt tgaggcgcta aatgaaacct taacgctatg gaactcgccg
    16981 cccgactggg ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc
    17041 gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat ggagcgcctg
    17101 ccggcccagt atcagcccgt catacttgaa gctagacagg cttatcttgg acaagaagaa
    17161 gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg tccactacgt gaaaggcgag
    17221 atcaccaagg tagtcggcaa ataatgtcta acaattcgtt caagccgacg ccgcttcgcg
    17281 gcgcggctta actcaagcgt tagatgcact aagcacataa ttgctcacag ccaaactatc
    17341 aggtcaagtc tgcttttatt atttttaagc gtgcataata agccctacac aaattgggag
    17401 atatatcatg aaaggctggc tttttcttgt tatcgcaata gttggcgaag taatcgcaac
    17461 atccgcatta aaatctagcg agggctttac taagctgatc cggtggatga ccttttgaat
    17521 gacctttaat agattatatt actaattaat tggggaccct agaggtcccc ttttttattt
    17581 taaaaatttt ttcacaaaac ggtttacaag cataaagctt gctcaatcaa tcaccggatc
    17641 cttaattaaa tcttccgctg cataaccctg cttcggggtc attatagcga ttttttcggt
    17701 atatccatcc tttttcgcac gatatacagg attttgccaa agggttcgtg tagactttcc
    17761 ttggtgtatc caacggcgtc agccgggcag gataggtgaa gtaggcccac ccgcgagcgg
    17821 gtgttccttc ttcactgtcc cttattcgca cctggcggtg ctcaacggga atcctgctct
    17881 gcgaggctgg ccggctaccg ccggcgtaac agatgagggc aagcggatgg ctgatgaaac
    17941 caagccaacc aggaagggca gcccacctat caaggtgtac tgccttccag acgaacgaag
    18001 agcgattgag gaaaaggcgg cggcggccgg catgagcctg tcggcctacc tgctggccgt
    18061 cggccagggc tacaaaatca cgggcgtcgt ggactatgag cacgtccgcg agctggcccg
    18121 catcaatggc gacctgggcc gcctgggcgg cctgctgaaa ctctggctca ccgacgaccc
    18181 gcgcacggcg cggttcggtg atgccacgat cctcgccctg ctggcgaaga tcgaagagaa
    18241 gcaggacgag cttggcaagg tcatgatggg cgtggtccgc ccgagggcag agccatgact
    18301 tttttagccg ctaaaacggc cggggggtgc gcgtgattgc caagcacgtc cccatgcgct
    18361 ccatcaagaa gaggcacttc gagctgtaag tacatcaccg acgagcaagg caagacgatc
//
