LOCUS       sequence-375867-       18477 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 Tet resistance..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 18477)
  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 18477)
  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: pAGE2.0
FEATURES             Location/Qualifiers
     source          1..18477
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            703..812
                     /note="incP origin of transfer"
     CDS             845..1216
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     CDS             1938..2507
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGFT
                     LREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNRR
                     LTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFTL
                     CGLDTALYDGTASDGEQALYMSMPCP"
     rep_origin      7242..7856
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     rep_origin      7333..7864
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      7932..8151
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             8242..8997
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     misc_feature    9000..9250
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     misc_feature    9412..9915
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        9916..10102
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             10103..10765
                     /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             10955..12130
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     CDS             12130..13101
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     misc_feature    13174..13647
                     /gene="sopC"
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     misc_feature    13907..14305
                     /label=cos
                     /note="lambda cos site; allows packaging into phage lambda 
                     particles"
     protein_bind    complement(14323..14356)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     14424..14443
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(14523..14798)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSYLGETIEYSSLHACRSTLEDPRVPSSNSPYSESYYNSL
                     AVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRS"
     primer_bind     14636..14658
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     14650..14667
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     14651..14667
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     14674..14693
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        14674..14692
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    14695..14751
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(14794..14810)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14794..14810)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(14807..14829)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    14818..14834
                     /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(14842..14848,14849..14866,14867..14872))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    14887..14908
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(15128..15787)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     15702..15721
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(15788..15890)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             complement(16930..17580)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     CDS             join(17686..18477,1..408)
                     /codon_start=1
                     /gene="tetA"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
ORIGIN
        1 atgatcaccg gccctgtagc cgcccggctc ggcgaaaggc gggcactcat gctcggaatg
       61 attgccgacg gcacaggcta catcctgctt gccttcgcga cacggggatg gatggcgttc
      121 ccgatcatgg tcctgcttgc ttcgggtggc atcggaatgc cggcgctgca agcaatgttg
      181 tccaggcagg tggatgagga acgtcagggg cagctgcaag gctcactggc ggcgctcacc
      241 agcctgacct cgatcgtcgg acccctcctc ttcacggcga tctatgcggc ttctataaca
      301 acgtggaacg ggtgggcatg gattgcaggc gctgccctct acttgctctg cctgccggcg
      361 ctgcgtcgcg ggctttggag cggcgcaggg caacgagccg atcgctgatc gtggaaacga
      421 taggcctatg ccatgcgggt caaggcgact tccggcaagc tatacgcgcc ctaggagtgc
      481 ggttggaacg ttggcccagc cagatactcc cgatcacgag caggacgccg atgatttgaa
      541 gcgcactcag cgtctgatct taattaaatc ttccgctgca taaccctgct tcggggtcat
      601 tatagcgatt ttttcggtat atccatcctt tttcgcacga tatacaggat tttgccaaag
      661 ggttcgtgta gactttcctt ggtgtatcca acggcgtcag ccgggcagga taggtgaagt
      721 aggcccaccc gcgagcgggt gttccttctt cactgtccct tattcgcacc tggcggtgct
      781 caacgggaat cctgctctgc gaggctggcc ggctaccgcc ggcgtaacag atgagggcaa
      841 gcggatggct gatgaaacca agccaaccag gaagggcagc ccacctatca aggtgtactg
      901 ccttccagac gaacgaagag cgattgagga aaaggcggcg gcggccggca tgagcctgtc
      961 ggcctacctg ctggccgtcg gccagggcta caaaatcacg ggcgtcgtgg actatgagca
     1021 cgtccgcgag ctggcccgca tcaatggcga cctgggccgc ctgggcggcc tgctgaaact
     1081 ctggctcacc gacgacccgc gcacggcgcg gttcggtgat gccacgatcc tcgccctgct
     1141 ggcgaagatc gaagagaagc aggacgagct tggcaaggtc atgatgggcg tggtccgccc
     1201 gagggcagag ccatgacttt tttagccgct aaaacggccg gggggtgcgc gtgattgcca
     1261 agcacgtccc catgcgctcc atcaagaaga ggcacttcga gctgtaagta catcaccgac
     1321 gagcaaggca agacgatcat cgatactagc ttgattggga tatctcgctc gtgcttgtcg
     1381 cgtgctatgt cttttagggt actttgaacc tacgttcgta cttgtataat atgatcatcg
     1441 tattatcgtt tttcatccgt ccagcgcaaa atgcattagc agctagtcct agcgtgcgga
     1501 gctacctgga caggtgcatg acggatgcgt gtccttcagt gactttctaa ttaacagtaa
     1561 cttctttact tatgtttcag tttgtaagaa gcgggattcg ctcgtcggtt gacatctgat
     1621 tggactgcgt cggcacatga aaactacatt gtgaaatctg ctaaaactcc gggtatctct
     1681 gacacaaaac gattcggctt cgcaatttca acattacggt caaggctaac gtatctttct
     1741 cggtcaactt cagattatgc cgattaaatt gtcgtagctt tcaaggcgtt ttgagtactg
     1801 cggcagttgt tgaacctgca aggagaagat ctcgacaaca gaataaagcg aaaaatgggt
     1861 ctcatgcact aacactcagg cctccctcat aatctctgtt tgagtttacc aacaacacat
     1921 atatacattt cgacaaaatg accactcttg acgacacggc ttaccggtac cgcaccagtg
     1981 tcccggggga cgccgaggcc atcgaggcac tggatgggtc cttcaccacc gacaccgtct
     2041 tccgcgtcac cgccaccggg gacggcttca ccctgcggga ggtgccggtg gacccgcccc
     2101 tgaccaaggt gttccccgac gacgaatcgg acgacgaatc ggacgacggg gaggacggcg
     2161 acccggactc ccggacgttc gtcgcgtacg gggacgacgg cgacctggcg ggcttcgtgg
     2221 tcgtctcgta ctccggctgg aaccgccggc tgaccgtcga ggacatcgag gtcgccccgg
     2281 agcaccgggg gcacggggtc gggcgcgcgt tgatggggct cgcgacggag ttcgcccgcg
     2341 agcggggcgc cgggcacctc tggctggagg tcaccaacgt caacgcaccg gcgatccacg
     2401 cgtaccggcg gatggggttc accctctgcg gcctggacac cgccctgtac gacggcaccg
     2461 cctcggacgg cgagcaggcg ctctacatga gcatgccctg cccctgattt tgttacattt
     2521 actgacttca aggagtcgag gaatcgatac tgccgtcgtt tccaggatcc gaggtttcat
     2581 aaactctgtt aacgttatag aaacagactt acctctccta cgccattcac gtaatattcg
     2641 caatatgcta ttcttcctct gaagaccagg tttatgtgct gcctgaaact atttcaataa
     2701 gtcagctgca cttgcacagg gtttcacaag gaaagcgtgt ctttttttcc aacgtaggcg
     2761 tcgctttcgt ctgactctta ctcttacatt cacagccaat acttacaatt agtaaaaaac
     2821 ctgtgctcga gagtgaaaac tccgtaacta taacggtcct aaggtagcga acactccgtt
     2881 cgtggcgtcg cacgatcgcg acatttgacg acgtgtgttt tgcctgggct acgtatcggg
     2941 aatgtttgcg aaacccccga aattgtaccg ggatgtaggc acatcgctgg aaggtagcca
     3001 ggggggtccg gcgctgatta cgaagtgcgg cgaccgacga atttggagca cgcccgacgc
     3061 tcaggggtgg atgccgctgg cgattccgct tgagtgggat cgctcggcac ccgcgggtcg
     3121 gcatatgccc gtgccgattg tgtccccacg gtgttggtca ggcggccggt atcacgacac
     3181 cccccgccca tcgaagccat tcgctctcag caacgccatg agcatcgggc caagcgagaa
     3241 tttcccttgc cgcgccttgg cggtcagatc gcgcagatag ccgccagcag agttgatatg
     3301 ccctgccctc tctagaatgc aggccacggc gactccggcg ttttccggtc ccatgacctc
     3361 gcaggcctcc tgataggccg aggggtaaac gccaagcata gatcgtaccg tgactgcagc
     3421 gatcatgagg tcgcgccaac ttctcactgc accgtcagga ccgtagttca tgatttccgg
     3481 gcaggctttc agtaccattc ccagcggaaa ggcccttttt gccttgttgg ccggttcaat
     3541 ggcttggccg tctaaccgct cattccgaga gcaacgttca agttcaataa tagattcggg
     3601 ttttgattcc tgtttgtgcc gctcagtttg atcgggattg ccgctcatct cgtgtggttt
     3661 tatgtgcttt tgcaactggt tagccacctg ttcgcgcagc gtttcgaatg ccactaccaa
     3721 aggggtgaga tcagcgatat cgggcgtcct cggtatcgcc tggacgaggc ttcggaaatc
     3781 ttcctgcata gacgtccagt ggcccggcac ttctacgagc accacgtcta tcaacttggc
     3841 gatgtctctg cggcagagcg tcagtcgctc gcgtagccgc tgcagctcca tccgatcagc
     3901 ctcggcttcg gcagcgagtc gctctatttc gccggagcgg gccagcagtg gtgcgagcga
     3961 aaagccgaac gctgtgccga tcgctccatc gcgagtcttg cgtgcatagc gcttgccgtt
     4021 cggactgtcc ttgcgagcga ccagccctgc atcgacgagg gcggccagat gccttctgag
     4081 cgtagcctcg gccatgccat gagctctcag tgacagctga acgttcgagg gaaagacgac
     4141 cagactggtc tccccgctta gcgtgttctc cggatagaag ctcaacagcg cgttcagaac
     4201 ggctagtgtc cgatcgctga tgccaagacg gcgtttggca gcacaaagcg agcgaaacaa
     4261 tttccacttg tcgaccgact tgccaggttc gatgtcgcgg gagatgaact ggcctgccaa
     4321 cagggcaagc gtcatcgacc gccgcccaaa gggcgtcgtc acacttccaa tctccatttc
     4381 cttcaccttc gataaggcaa aagaaagcag ctcaccaaaa acggtgccaa ggactcttga
     4441 ctgtgattcg tggaaatgcg attctcgatc ttgctacaaa tctgaggaag gcttccacgg
     4501 cttgttcgtt gggggccttt ttcttttgcg ggttagtctc cttgtttcga actctgcttt
     4561 tgcatgaact ggtcgtgcag gctttcgatc tgacctgcaa tgaaatcgac aaagcctgac
     4621 ggcacagact tgttgaatgt cagcttcgca gccgacttgc cgcgttggat ctggccaagc
     4681 acgaggccat gagagatcag atcgctcgtg gaagacgctg cagccctcac atcaaccttg
     4741 gtaagggctt caaaaaccgc agtgaagcgt agatcgctgg aagaggcttg aatctgctct
     4801 tgctccagaa ccaactgcgc caccttcgcg ggctcccggt gagccgccca gcgcacttcg
     4861 aactcgatcc atctacgccg tccgatttca ggagccggac caatctgcat caggatctcg
     4921 cgcggcaatg ctgccgctat ccggatcatt tcggagatat gcgacctgcc ggtgctcagg
     4981 gacttgcaga tcacgtcgcg gcgatagccg gcagcctcct gcgattgcgc aaaaaggcat
     5041 cgctcgatga aggacaggtc cttgcggttg ctgttctctt ccccttggag aacgatcgca
     5101 tcttcgtcgc taagttcccg ggcgattgcc ttgaacttga tgccgaggag tttggcagcg
     5161 gcaagccggc gccggccaaa tacgatctga aacggccttg cggcccctcg aaccgggcgg
     5221 accaagccag gagtactctg tccatgctcg cgcatcgact cgagcaaatc cgcaataccg
     5281 gcttcgctgt agccggaatc aaagcgatcc cggatggccg attccatgat ctcgtctgga
     5341 tcgatctcga cgatctggcc accatccttg agaaggcgtt cggcaagttc gctgcgttcc
     5401 tgcatctgac ggacgccggc tgcaactttg agaagatgtg gcgatgtcga ccgctgcatc
     5461 ccgcccttct ctgcagaatt ttccagctgt ccgaggacat tggcaaaaag gcccttggaa
     5521 tccttgcggc tcattctctc ccccatgcct tccagaagag gccctcgagt tcagcattgg
     5581 cagcattgat cgactcgatg gcgcggtcgt aggttttcgg cgccgaaaat cgggacctct
     5641 gaacctcgta aaggctctgc ttccacgtca gagcatcggc caccgcggtc gacttgatga
     5701 cggagttgag gagcagatcc tcgccgaaca cctgtctcat cagcgcggac atattcgctt
     5761 gcggatgatc gttcggctcg aactttgtga tcaggaactt ggcgaagtcc cattgagcgg
     5821 cggtgcccac ctcattcagg atctgcatgt acgacccggc aagctccaaa aactttgcgg
     5881 tcgagtccac atccagcata tgtgccggaa ccgggatgat gactccagtc gcggcggtca
     5941 gcgacgaaag tgtcaggaag ttcagcgacg gagccgagtc cataaggacg aagtcgtagt
     6001 catcggcgac ctgctccaga gcttgagaaa cgcgaagcaa gaaaccggtt cctgctgacc
     6061 gccgcatctc gagggcgacg gccgtctcga actcggtcag ttcgaggccg gcacagatga
     6121 cgtcgaaacc aacgatgtgg gtcttgtgga tgatggattt cgtcgggacg ggatcgtcga
     6181 aacgaatggc tgagaaaagt gtatcgccgc cgcggtaatc gaattcgggc aggctaccat
     6241 ggagcgcggt gagagatgct tgcgggtcga gatcgaccgc cagcacgcga tagcccttga
     6301 gtgccagata gtgaccgaga tggattgtgg tagaggtctt gctggagccg cccttgaagt
     6361 tggtgaccgc aacgatctgg cattgctcat tgcctgtccg gcgggggttc atccatttct
     6421 tccggccgtt tgccgcaaga gccatacgta gctcaagaac ctgctcgagc gtgtagaagc
     6481 gccggccgtt ggcgatttcg ggctcagggc cctgttcctt ggcggcaacc tggcggatat
     6541 aggcctcggt tacgcccagc agggtggcaa cttcggtaga ggtgaacttg cgcattgtgc
     6601 gacgcgcgtt cggggggaac tgcgccatgc tcaagttgtt cagtgccatt tcgagcttac
     6661 tggaaaaatt ctccatgaaa gcgaggctgc ttaggtgtcg cgtcatcgaa agaatccctg
     6721 tgatcgtttc cgtcacttaa gcagtgaagt ggttaaccgc ttgctaatcg ccacgtaact
     6781 actcgaaaac acgcaaattc gaaaaatttc cgcaactaag acgtactccg attcggctat
     6841 ggccatcaat gcgcgctgtg gatgacgcac agagccgggg ggaaggcgcc agtgcatgcc
     6901 ggttctcgac cctccccgcc tcttttttgg acgaagatgt gcgcagacgt cgctgggcgg
     6961 agacgcaata gtggttcact gcggaaatgg ttggccgttg ctttcaatgg agatctgccg
     7021 aactcttcgc agagttcccc gcggggaact ctcgaagcga cgacgctggc gcgcagtacc
     7081 gactgctcgc caaggcgaga catatgtgcc gtattgggaa cagctcggca tctcggtgct
     7141 cgcctgcgct ccggcctgcc tcaatctcgt tggccatgac cgagcaacga ctgcaccgga
     7201 tggttcagtt acgctaggga taacagggta atatagtcga gctggttgcc ctcgccgctg
     7261 ggctggcggc cgtctatggc cctgcaaacg cgccagaaac gccgtcgaag ccgtgtgcga
     7321 gacaccgcgg ccggccgccg gcgttgtgga tacctcgcgg aaaacttggc cctcactgac
     7381 agatgagggg cggacgttga cacttgaggg gccgactcac ccggcgcggc gttgacagat
     7441 gaggggcagg ctcgatttcg gccggcgacg tggagctggc cagcctcgca aatcggcgaa
     7501 aacgcctgat tttacgcgag tttcccacag atgatgtgga caagcctggg gataagtgcc
     7561 ctgcggtatt gacacttgag gggcgcgact actgacagat gaggggcgcg atccttgaca
     7621 cttgaggggc agagtgctga cagatgaggg gcgcacctat tgacatttga ggggctgtcc
     7681 acaggcagaa aatccagcat ttgcaagggt ttccgcccgt ttttcggcca ccgctaacct
     7741 gtcttttaac ctgcttttaa accaatattt ataaaccttg tttttaacca gggctgcgcc
     7801 ctgtgcgcgt gaccgcgcac gccgaagggg ggtgcccccc cttctcgaac cctcccggtc
     7861 gagtgagcga ggaagcacca gggaacagca cttatatatt ctgcttacac acgatgcctg
     7921 aaaaaacttc ccttggggtt atccacttat ccacggggat atttttataa ttattttttt
     7981 tatagttttt agatcttctt ttttagagcg ccttgtaggc ctttatccat gctggttcta
     8041 gagaaggtgt tgtgacaaat tgccctttca gtgtgacaaa tcaccctcaa atgacagtcc
     8101 tgtctgtgac aaattgccct taaccctgtg acaaattgcc ctcagaagaa gctgtttttt
     8161 cacaaagtta tccctgctta ttgactcttt tttatttagt gtgacaatct aaaaacttgt
     8221 cacacttcac atggatctgt catggcggaa acagcggtta tcaatcacaa gaaacgtaaa
     8281 aatagcccgc gaatcgtcca gtcaaacgac ctcactgagg cggcatatag tctctcccgg
     8341 gatcaaaaac gtatgctgta tctgttcgtt gaccagatca gaaaatctga tggcacccta
     8401 caggaacatg acggtatctg cgagatccat gttgctaaat atgctgaaat attcggattg
     8461 acctctgcgg aagccagtaa ggatatacgg caggcattga agagtttcgc ggggaaggaa
     8521 gtggtttttt atcgccctga agaggatgcc ggcgatgaaa aaggctatga atcttttcct
     8581 tggtttatca aacgtgcgca cagtccatcc agagggcttt acagtgtaca tatcaaccca
     8641 tatctcattc ccttctttat cgggttacag aaccggttta cgcagtttcg gcttagtgaa
     8701 acaaaagaaa tcaccaatcc gtatgccatg cgtttatacg aatccctgtg tcagtatcgt
     8761 aagccggatg gctcaggcat cgtctctctg aaaatcgact ggatcataga gcgttaccag
     8821 ctgcctcaaa gttaccagcg tatgcctgac ttccgccgcc gcttcctgca ggtctgtgtt
     8881 aatgagatca acagcagaac tccaatgcgc ctctcataca ttgagaaaaa gaaaggccgc
     8941 cagacgactc atatcgtatt ttccttccgc gatatcactt ccatgacgac aggatagtct
     9001 gagggttatc tgtcacagat ttgagggtgg ttcgtcacat ttgttctgac ctactgaggg
     9061 taatttgtca cagttttgct gtttccttca gcctgcatgg attttctcat actttttgaa
     9121 ctgtaatttt taaggaagcc aaatttgagg gcagtttgtc acagttgatt tccttctctt
     9181 tcccttcgtc atgtgacctg atatcggggg ttagttcgtc atcattgatg agggttgatt
     9241 atcacagttt attactctga attggctatc cgcgtgtgta cctctacctg gagtttttcc
     9301 cacggtggat atttcttctt gcgctgagcg taagagctat ctgacagaac agttcttctt
     9361 tgcttcctcg ccagttcgct cgctatgctc ggttacacgg ctgcggcgag catcacgtgc
     9421 tataaaaata attataattt aaatttttta atataaatat ataaattaaa aatagaaagt
     9481 aaaaaaagaa attaaagaaa aaatagtttt tgttttccga agatgtaaaa gactctaggg
     9541 ggatcgccaa caaatactac cttttacctt gctcttcctg ctctcaggta ttaatgccga
     9601 attgtttcat cttgtctgtg tagaagacca cacacgaaaa tcctgtgatt ttacatttta
     9661 cttatcgtta atcgaatgta tatctattta atctgctttt cttgtctaat aaatatatat
     9721 gtaaagtacg ctttttgttg aaatttttta aacctttgtt tatttttttt tcttcattcc
     9781 gtaactcttc taccttcttt atttactttc taaaatccaa atacaaaaca taaaaataaa
     9841 taaacacaga gtaaattccc aaattattcc atcattaaaa gatacgaggc gcgtgtaagt
     9901 tacaggcaag cgatcctagt acactctata tttttttatg cctcggtaat gattttcatt
     9961 ttttttttcc acctagcgga tgactctttt tttttcttag cgattggcat tatcacataa
    10021 tgaattatac attatataaa gtaatgtgat ttcttcgaag aatatactaa aaaatgagca
    10081 ggcaagataa acgaaggcaa agatgacaga gcagaaagcc ctagtaaagc gtattacaaa
    10141 tgaaaccaag attcagattg cgatctcttt aaagggtggt cccctagcga tagagcactc
    10201 gatcttccca gaaaaagagg cagaagcagt agcagaacag gccacacaat cgcaagtgat
    10261 taacgtccac acaggtatag ggtttctgga ccatatgata catgctctgg ccaagcattc
    10321 cggctggtcg ctaatcgttg agtgcattgg tgacttacac atagacgacc atcacaccac
    10381 tgaagactgc gggattgctc tcggtcaagc ttttaaagag gccctagggg ccgtgcgtgg
    10441 agtaaaaagg tttggatcag gatttgcgcc tttggatgag gcactttcca gagcggtggt
    10501 agatctttcg aacaggccgt acgcagttgt cgaacttggt ttgcaaaggg agaaagtagg
    10561 agatctctct tgcgagatga tcccgcattt tcttgaaagc tttgcagagg ctagcagaat
    10621 taccctccac gttgattgtc tgcgaggcaa gaatgatcat caccgtagtg agagtgcgtt
    10681 caaggctctt gcggttgcca taagagaagc cacctcgccc aatggtacca acgatgttcc
    10741 ctccaccaaa ggtgttctta tgtagtttta cacaggagtc tggacttgac gctagtgata
    10801 ataagtgact gaggtatgtg ctcttcttat ctccttttgt agtgttgctc ttattttaaa
    10861 caactttgcg gttttttgat gactttgcga ttttgttgtt gctttgcagt aaattgcaag
    10921 atttaataaa aaaacgcaaa gcaatgatta aaggatgttc agaatgaaac tcatggaaac
    10981 acttaaccag tgcataaacg ctggtcatga aatgacgaag gctatcgcca ttgcacagtt
    11041 taatgatgac agcccggaag cgaggaaaat aacccggcgc tggagaatag gtgaagcagc
    11101 ggatttagtt ggggtttctt ctcaggctat cagagatgcc gagaaagcag ggcgactacc
    11161 gcacccggat atggaaattc gaggacgggt tgagcaacgt gttggttata caattgaaca
    11221 aattaatcat atgcgtgatg tgtttggtac gcgattgcga cgtgctgaag acgtatttcc
    11281 accggtgatc ggggttgctg cccataaagg tggcgtttac aaaacctcag tttctgttca
    11341 tcttgctcag gatctggctc tgaaggggct acgtgttttg ctcgtggaag gtaacgaccc
    11401 ccagggaaca gcctcaatgt atcacggatg ggtaccagat cttcatattc atgcagaaga
    11461 cactctcctg cctttctatc ttggggaaaa ggacgatgtc acttatgcaa taaagcccac
    11521 ttgctggccg gggcttgaca ttattccttc ctgtctggct ctgcaccgta ttgaaactga
    11581 gttaatgggc aaatttgatg aaggtaaact gcccaccgat ccacacctga tgctccgact
    11641 ggccattgaa actgttgctc atgactatga tgtcatagtt attgacagcg cgcctaacct
    11701 gggtatcggc acgattaatg tcgtatgtgc tgctgatgtg ctgattgttc ccacgcctgc
    11761 tgagttgttt gactacacct ccgcactgca gtttttcgat atgcttcgtg atctgctcaa
    11821 gaacgttgat cttaaagggt tcgagcctga tgtacgtatt ttgcttacca aatacagcaa
    11881 tagcaatggc tctcagtccc cgtggatgga ggagcaaatt cgggatgcct ggggaagcat
    11941 ggttctaaaa aatgttgtac gtgaaacgga tgaagttggt aaaggtcaga tccggatgag
    12001 aactgttttt gaacaggcca ttgatcaacg ctcttcaact ggtgcctgga gaaatgctct
    12061 ttctatttgg gaacctgtct gcaatgaaat tttcgatcgt ctgattaaac cacgctggga
    12121 gattagataa tgaagcgtgc gcctgttatt ccaaaacata cgctcaatac tcaaccggtt
    12181 gaagatactt cgttatcgac accagctgcc ccgatggtgg attcgttaat tgcgcgcgta
    12241 ggagtaatgg ctcgcggtaa tgccattact ttgcctgtat gtggtcggga tgtgaagttt
    12301 actcttgaag tgctccgggg tgatagtgtt gagaagacct ctcgggtatg gtcaggtaat
    12361 gaacgtgacc aggagctgct tactgaggac gcactggatg atctcatccc ttcttttcta
    12421 ctgactggtc aacagacacc ggcgttcggt cgaagagtat ctggtgtcat agaaattgcc
    12481 gatgggagtc gccgtcgtaa agctgctgca cttaccgaaa gtgattatcg tgttctggtt
    12541 ggcgagctgg atgatgagca gatggctgca ttatccagat tgggtaacga ttatcgccca
    12601 acaagtgctt atgaacgtgg tcagcgttat gcaagccgat tgcagaatga atttgctgga
    12661 aatatttctg cgctggctga tgcggaaaat atttcacgta agattattac ccgctgtatc
    12721 aacaccgcca aattgcctaa atcagttgtt gctctttttt ctcaccccgg tgaactatct
    12781 gcccggtcag gtgatgcact tcaaaaagcc tttacagata aagaggaatt acttaagcag
    12841 caggcatcta accttcatga gcagaaaaaa gctggggtga tatttgaagc tgaagaagtt
    12901 atcactcttt taacttctgt gcttaaaacg tcatctgcat caagaactag tttaagctca
    12961 cgacatcagt ttgctcctgg agcgacagta ttgtataagg gcgataaaat ggtgcttaac
    13021 ctggacaggt ctcgtgttcc aactgagtgt atagagaaaa ttgaggccat tcttaaggaa
    13081 cttgaaaagc cagcaccctg atgcgacctc gttttagtct acgtttatct gtctttactt
    13141 aatgtccttt gttacaggcc agaaagcata actggcctga atattctctc tgggcccact
    13201 gttccacttg tatcgtcggt ctgataatca gactgggacc acggtcccac tcgtatcgtc
    13261 ggtctgatta ttagtctggg accacggtcc cactcgtatc gtcggtctga ttattagtct
    13321 gggaccacgg tcccactcgt atcgtcggtc tgataatcag actgggacca cggtcccact
    13381 cgtatcgtcg gtctgattat tagtctggga ccatggtccc actcgtatcg tcggtctgat
    13441 tattagtctg ggaccacggt cccactcgta tcgtcggtct gattattagt ctggaaccac
    13501 ggtcccactc gtatcgtcgg tctgattatt agtctgggac cacggtccca ctcgtatcgt
    13561 cggtctgatt attagtctgg gaccacgatc ccactcgtgt tgtcggtctg attatcggtc
    13621 tgggaccacg gtcccacttg tattgtcgat cagactatca gcgtgagact acgattccat
    13681 caatgcctgt caagggcaag tattgacatg tcgtcgtaac ctgtagaacg gagtaacctc
    13741 ggtgtgcggt tgtatgcctg ctgtggattg ctgctgtgtc ctgcttatcc acaacatttt
    13801 gcgcacggtt atgtggacaa aatacctggt tacccaggcc gtgccggcac gttaaccggg
    13861 ctgcatccga tgcaagtgtg tcgctgtcga cgagctcgcg agctcggaca tgaggttgcc
    13921 ccgtattcag tgtcgctgat ttgtattgtc tgaagttgtt tttacgttaa gttgatgcag
    13981 atcaattaat acgatacctg cgtcataatt gattatttga cgtggtttga tggcctccac
    14041 gcacgttgtg atatgtagat gataatcatt atcactttac gggtcctttc cggtgatccg
    14101 acaggttacg gggcggcgac ctcgcgggtt ttcgctattt atgaaaattt tccggtttaa
    14161 ggcgtttccg ttcttcttcg tcataactta atgtttttat ttaaaatacc ctctgaaaag
    14221 aaaggaaacg acaggtgctg aaagcgagct ttttggcctc tgtcgtttcc tttctctgtt
    14281 tttgtccgtg gaatgaacaa tggaagtccg agctcatcgc taataacttc gtatagcata
    14341 cattatacga agttatattc gatgcggccg caaggggttc gcgtcagcgg gtgttggcgg
    14401 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
    14461 gcggtgtgaa ataccacaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
    14521 attcagctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
    14581 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
    14641 tcacgacgtt gtaaaacgac ggccagtgaa ttgtaatacg actcactata gggcgaattc
    14701 gagctcggta cccggggatc ctctagagtc gacctgcagg catgcaagct tgagtattct
    14761 atagtctcac ctaaatagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
    14821 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
    14881 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
    14941 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgaacccc ttgcggccgc
    15001 ccgggccgtc gaccaattct catgtttgac agcttatcat cgaatttctg ccattcatcc
    15061 gcttattatc acttattcag gcgtagcaac caggcgttta agggcaccaa taactgcctt
    15121 aaaaaaatta cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc
    15181 tgccgacatg gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca
    15241 ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca
    15301 tattggccac gtttaaatca aaactggtga aactcaccca gggattggct gagacgaaaa
    15361 acatattctc aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat
    15421 cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg
    15481 aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca
    15541 ccagctcacc gtctttcatt gccatacgaa attccggatg agcattcatc aggcgggcaa
    15601 gaatgtgaat aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg
    15661 ccgtaatatc cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct
    15721 caaaatgttc tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt
    15781 tctccatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta
    15841 gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgccgatca acgtctcatt
    15901 ttcgccaaaa gttggcccag ggcttcccgg tatcaacagg gacaccagga tttatttatt
    15961 ctgcgaagtg atcttccgtc acaggtattt attcgcgata agctcatgga gcggcgtaac
    16021 cgtcgcacag gaaggacaga gaaagcgcgg atctgggaag tgacggacag aacggtcagg
    16081 acctggattg gggaggcggt tgccgccgct gctgctgacg gtgtgacgtt ctctgttccg
    16141 gtcacaccac atacgttccg ccattcctat gcgatgcaca tgctgtatgc cggtataccg
    16201 ctgaaagttc tgcaaagcct gatgggacat aagtccatca gttcaacgga agtctacacg
    16261 aaggtttttg cgctggatgt ggctgcccgg caccgggtgc agtttgcgat gccggagtct
    16321 gatgcggttg cgatgctgaa acaattatcc tgagaataaa tgccttggcc tttatatgga
    16381 aatgtggaac tgagtggata tgctgttttt gtctgttaaa cagagaagct ggctgttatc
    16441 cactgagaag cgaacgaaac agtcgggaaa atctcccatt atcgtagaga tccgcattat
    16501 taatctcagg agcctgtgta gcgtttatag gaagtagtgt tctgtcatga tgcctgcaag
    16561 cggtaacgaa aacgatttga atatgccttc aggaacaata gaaatcttcg tgcggtgtta
    16621 cgttgaagtg gagcggatta tgtcagcaat ggacagaaca acctaatgaa cacagaacca
    16681 tgatgtggtc tgtcctttta cagccagtag tgctcgccgc agtcgagcga cagggcgaag
    16741 cccgtttaaa cgagcgccag aaggccgcca gagaggccga gcgcggccgt gaggcttgga
    16801 cgctagggca gggcatgaaa aagcccgtag cgggctgcta cgggcgtctg acgcggtgga
    16861 aagggggagg ggatgttgtc tacatggctc tgctgtagtg agtgggttgc gctccggcag
    16921 cggtcctgat caatcgtcac cctttctcgg tccttcaacg ttcctgacaa cgagcctcct
    16981 tttcgccaat ccatcgacaa tcaccgcgag tccctgctcg aacgctgcgt ccggaccggc
    17041 ttcgtcgaag gcgtctatcg cggcccgcaa cagcggcgag agcggagcct gttcaacggt
    17101 gccgccgcgc tcgccggcat cgctgtcgcc ggcctgctcc tcaagcacgg ccccaacagt
    17161 gaagtagctg attgtcatca gcgcattgac ggcgtccccg gccgaaaaac ccgcctcgca
    17221 gaggaagcga agctgcgcgt cggccgtttc catctgcggt gcgcccggtc gcgtgccggc
    17281 atggatgcgc gcgccatcgc ggtaggcgag cagcgcctgc ctgaagctgc gggcattccc
    17341 gatcagaaat gagcgccagt cgtcgtcggc tctcggcacc gaatgcgtat gattctccgc
    17401 cagcatggct tcggccagtg cgtcgagcag cgcccgcttg ttcctgaagt gccagtaaag
    17461 cgccggctgc tgaaccccca accgttccgc cagtttgcgt gtcgtcagac cgtctacgcc
    17521 gacctcgttc aacaggtcca gggcggcacg gatcactgta ttcggctgca actttgtcat
    17581 gcttgacact ttatcactga taaacataat atgtccacca acttatcagt gataaagaat
    17641 ccgcgcgttc aatcggacca gcggaggctg gtccggaggc cagacgtgaa acccaacata
    17701 cccctgatcg taattctgag cactgtcgcg ctcgacgctg tcggcatcgg cctgattatg
    17761 ccggtgctgc cgggcctcct gcgcgatctg gttcactcga acgacgtcac cgcccactat
    17821 ggcattctgc tggcgctgta tgcgttggtg caatttgcct gcgcacctgt gctgggcgcg
    17881 ctgtcggatc gtttcgggcg gcggccaatc ttgctcgtct cgctggccgg cgccactgtc
    17941 gactacgcca tcatggcgac agcgcctttc ctttgggttc tctatatcgg gcggatcgtg
    18001 gccggcatca ccggggcgac tggggcggta gccggcgctt atattgccga tatcactgat
    18061 ggcgatgagc gcgcgcggca cttcggcttc atgagcgcct gtttcgggtt cgggatggtc
    18121 gcgggacctg tgctcggtgg gctgatgggc ggtttctccc cccacgctcc gttcttcgcc
    18181 gcggcagcct tgaacggcct caatttcctg acgggctgtt tccttttgcc ggagtcgcac
    18241 aaaggcgaac gccggccgtt acgccgggag gctctcaacc cgctcgcttc gttccggtgg
    18301 gcccggggca tgaccgtcgt cgccgccctg atggcggtct tcttcatcat gcaacttgtc
    18361 ggacaggtgc cggccgcgct ttgggtcatt ttcggcgagg atcgctttca ctgggacgcg
    18421 accacgatcg gcatttcgct tgccgcattt ggcattctgc attcactcgc ccaggca
//
