LOCUS       sequence-474332-       16328 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transmissible episome that contains the NAT cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16328)
  TITLE     Spheroplasted cells: a game changer for DNA delivery to diatoms
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 16328)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pPtGE31
FEATURES             Location/Qualifiers
     source          1..16328
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      6..620
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     rep_origin      97..628
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      696..915
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             1006..1761
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     misc_feature    1764..2014
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     misc_feature    2176..2679
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        2680..2866
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             2867..3529
                     /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             3719..4894
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     CDS             4894..5865
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     misc_feature    5938..6411
                     /gene="sopC"
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     misc_feature    6671..7069
                     /label=cos
                     /note="lambda cos site; allows packaging into phage lambda 
                     particles"
     protein_bind    complement(7087..7120)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     7188..7207
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(7287..7562)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSYLGETIEYSSLHACRSTLEDPRVPSSNSPYSESYYNSL
                     AVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRS"
     primer_bind     7400..7422
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7414..7431
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7415..7431
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7438..7457
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7438..7456
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    7459..7515
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(7558..7574)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7558..7574)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7571..7593)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7582..7598
                     /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(7606..7612,7613..7630,7631..7636))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7651..7672
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(7892..8551)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     8466..8485
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(8552..8654)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             10107..10676
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGFT
                     LREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNRR
                     LTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFTL
                     CGLDTALYDGTASDGEQALYMSMPCP"
     promoter        11373..11588
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             11589..12392
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLATGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     oriT            13149..13258
                     /note="incP origin of transfer"
     CDS             13291..13662
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
ORIGIN
        1 tcgagctggt tgccctcgcc gctgggctgg cggccgtcta tggccctgca aacgcgccag
       61 aaacgccgtc gaagccgtgt gcgagacacc gcggccggcc gccggcgttg tggatacctc
      121 gcggaaaact tggccctcac tgacagatga ggggcggacg ttgacacttg aggggccgac
      181 tcacccggcg cggcgttgac agatgagggg caggctcgat ttcggccggc gacgtggagc
      241 tggccagcct cgcaaatcgg cgaaaacgcc tgattttacg cgagtttccc acagatgatg
      301 tggacaagcc tggggataag tgccctgcgg tattgacact tgaggggcgc gactactgac
      361 agatgagggg cgcgatcctt gacacttgag gggcagagtg ctgacagatg aggggcgcac
      421 ctattgacat ttgaggggct gtccacaggc agaaaatcca gcatttgcaa gggtttccgc
      481 ccgtttttcg gccaccgcta acctgtcttt taacctgctt ttaaaccaat atttataaac
      541 cttgttttta accagggctg cgccctgtgc gcgtgaccgc gcacgccgaa ggggggtgcc
      601 cccccttctc gaaccctccc ggtcgagtga gcgaggaagc accagggaac agcacttata
      661 tattctgctt acacacgatg cctgaaaaaa cttcccttgg ggttatccac ttatccacgg
      721 ggatattttt ataattattt tttttatagt ttttagatct tcttttttag agcgccttgt
      781 aggcctttat ccatgctggt tctagagaag gtgttgtgac aaattgccct ttcagtgtga
      841 caaatcaccc tcaaatgaca gtcctgtctg tgacaaattg cccttaaccc tgtgacaaat
      901 tgccctcaga agaagctgtt ttttcacaaa gttatccctg cttattgact cttttttatt
      961 tagtgtgaca atctaaaaac ttgtcacact tcacatggat ctgtcatggc ggaaacagcg
     1021 gttatcaatc acaagaaacg taaaaatagc ccgcgaatcg tccagtcaaa cgacctcact
     1081 gaggcggcat atagtctctc ccgggatcaa aaacgtatgc tgtatctgtt cgttgaccag
     1141 atcagaaaat ctgatggcac cctacaggaa catgacggta tctgcgagat ccatgttgct
     1201 aaatatgctg aaatattcgg attgacctct gcggaagcca gtaaggatat acggcaggca
     1261 ttgaagagtt tcgcggggaa ggaagtggtt ttttatcgcc ctgaagagga tgccggcgat
     1321 gaaaaaggct atgaatcttt tccttggttt atcaaacgtg cgcacagtcc atccagaggg
     1381 ctttacagtg tacatatcaa cccatatctc attcccttct ttatcgggtt acagaaccgg
     1441 tttacgcagt ttcggcttag tgaaacaaaa gaaatcacca atccgtatgc catgcgttta
     1501 tacgaatccc tgtgtcagta tcgtaagccg gatggctcag gcatcgtctc tctgaaaatc
     1561 gactggatca tagagcgtta ccagctgcct caaagttacc agcgtatgcc tgacttccgc
     1621 cgccgcttcc tgcaggtctg tgttaatgag atcaacagca gaactccaat gcgcctctca
     1681 tacattgaga aaaagaaagg ccgccagacg actcatatcg tattttcctt ccgcgatatc
     1741 acttccatga cgacaggata gtctgagggt tatctgtcac agatttgagg gtggttcgtc
     1801 acatttgttc tgacctactg agggtaattt gtcacagttt tgctgtttcc ttcagcctgc
     1861 atggattttc tcatactttt tgaactgtaa tttttaagga agccaaattt gagggcagtt
     1921 tgtcacagtt gatttccttc tctttccctt cgtcatgtga cctgatatcg ggggttagtt
     1981 cgtcatcatt gatgagggtt gattatcaca gtttattact ctgaattggc tatccgcgtg
     2041 tgtacctcta cctggagttt ttcccacggt ggatatttct tcttgcgctg agcgtaagag
     2101 ctatctgaca gaacagttct tctttgcttc ctcgccagtt cgctcgctat gctcggttac
     2161 acggctgcgg cgagcatcac gtgctataaa aataattata atttaaattt tttaatataa
     2221 atatataaat taaaaataga aagtaaaaaa agaaattaaa gaaaaaatag tttttgtttt
     2281 ccgaagatgt aaaagactct agggggatcg ccaacaaata ctacctttta ccttgctctt
     2341 cctgctctca ggtattaatg ccgaattgtt tcatcttgtc tgtgtagaag accacacacg
     2401 aaaatcctgt gattttacat tttacttatc gttaatcgaa tgtatatcta tttaatctgc
     2461 ttttcttgtc taataaatat atatgtaaag tacgcttttt gttgaaattt tttaaacctt
     2521 tgtttatttt tttttcttca ttccgtaact cttctacctt ctttatttac tttctaaaat
     2581 ccaaatacaa aacataaaaa taaataaaca cagagtaaat tcccaaatta ttccatcatt
     2641 aaaagatacg aggcgcgtgt aagttacagg caagcgatcc tagtacactc tatatttttt
     2701 tatgcctcgg taatgatttt catttttttt ttccacctag cggatgactc tttttttttc
     2761 ttagcgattg gcattatcac ataatgaatt atacattata taaagtaatg tgatttcttc
     2821 gaagaatata ctaaaaaatg agcaggcaag ataaacgaag gcaaagatga cagagcagaa
     2881 agccctagta aagcgtatta caaatgaaac caagattcag attgcgatct ctttaaaggg
     2941 tggtccccta gcgatagagc actcgatctt cccagaaaaa gaggcagaag cagtagcaga
     3001 acaggccaca caatcgcaag tgattaacgt ccacacaggt atagggtttc tggaccatat
     3061 gatacatgct ctggccaagc attccggctg gtcgctaatc gttgagtgca ttggtgactt
     3121 acacatagac gaccatcaca ccactgaaga ctgcgggatt gctctcggtc aagcttttaa
     3181 agaggcccta ggggccgtgc gtggagtaaa aaggtttgga tcaggatttg cgcctttgga
     3241 tgaggcactt tccagagcgg tggtagatct ttcgaacagg ccgtacgcag ttgtcgaact
     3301 tggtttgcaa agggagaaag taggagatct ctcttgcgag atgatcccgc attttcttga
     3361 aagctttgca gaggctagca gaattaccct ccacgttgat tgtctgcgag gcaagaatga
     3421 tcatcaccgt agtgagagtg cgttcaaggc tcttgcggtt gccataagag aagccacctc
     3481 gcccaatggt accaacgatg ttccctccac caaaggtgtt cttatgtagt tttacacagg
     3541 agtctggact tgacgctagt gataataagt gactgaggta tgtgctcttc ttatctcctt
     3601 ttgtagtgtt gctcttattt taaacaactt tgcggttttt tgatgacttt gcgattttgt
     3661 tgttgctttg cagtaaattg caagatttaa taaaaaaacg caaagcaatg attaaaggat
     3721 gttcagaatg aaactcatgg aaacacttaa ccagtgcata aacgctggtc atgaaatgac
     3781 gaaggctatc gccattgcac agtttaatga tgacagcccg gaagcgagga aaataacccg
     3841 gcgctggaga ataggtgaag cagcggattt agttggggtt tcttctcagg ctatcagaga
     3901 tgccgagaaa gcagggcgac taccgcaccc ggatatggaa attcgaggac gggttgagca
     3961 acgtgttggt tatacaattg aacaaattaa tcatatgcgt gatgtgtttg gtacgcgatt
     4021 gcgacgtgct gaagacgtat ttccaccggt gatcggggtt gctgcccata aaggtggcgt
     4081 ttacaaaacc tcagtttctg ttcatcttgc tcaggatctg gctctgaagg ggctacgtgt
     4141 tttgctcgtg gaaggtaacg acccccaggg aacagcctca atgtatcacg gatgggtacc
     4201 agatcttcat attcatgcag aagacactct cctgcctttc tatcttgggg aaaaggacga
     4261 tgtcacttat gcaataaagc ccacttgctg gccggggctt gacattattc cttcctgtct
     4321 ggctctgcac cgtattgaaa ctgagttaat gggcaaattt gatgaaggta aactgcccac
     4381 cgatccacac ctgatgctcc gactggccat tgaaactgtt gctcatgact atgatgtcat
     4441 agttattgac agcgcgccta acctgggtat cggcacgatt aatgtcgtat gtgctgctga
     4501 tgtgctgatt gttcccacgc ctgctgagtt gtttgactac acctccgcac tgcagttttt
     4561 cgatatgctt cgtgatctgc tcaagaacgt tgatcttaaa gggttcgagc ctgatgtacg
     4621 tattttgctt accaaataca gcaatagcaa tggctctcag tccccgtgga tggaggagca
     4681 aattcgggat gcctggggaa gcatggttct aaaaaatgtt gtacgtgaaa cggatgaagt
     4741 tggtaaaggt cagatccgga tgagaactgt ttttgaacag gccattgatc aacgctcttc
     4801 aactggtgcc tggagaaatg ctctttctat ttgggaacct gtctgcaatg aaattttcga
     4861 tcgtctgatt aaaccacgct gggagattag ataatgaagc gtgcgcctgt tattccaaaa
     4921 catacgctca atactcaacc ggttgaagat acttcgttat cgacaccagc tgccccgatg
     4981 gtggattcgt taattgcgcg cgtaggagta atggctcgcg gtaatgccat tactttgcct
     5041 gtatgtggtc gggatgtgaa gtttactctt gaagtgctcc ggggtgatag tgttgagaag
     5101 acctctcggg tatggtcagg taatgaacgt gaccaggagc tgcttactga ggacgcactg
     5161 gatgatctca tcccttcttt tctactgact ggtcaacaga caccggcgtt cggtcgaaga
     5221 gtatctggtg tcatagaaat tgccgatggg agtcgccgtc gtaaagctgc tgcacttacc
     5281 gaaagtgatt atcgtgttct ggttggcgag ctggatgatg agcagatggc tgcattatcc
     5341 agattgggta acgattatcg cccaacaagt gcttatgaac gtggtcagcg ttatgcaagc
     5401 cgattgcaga atgaatttgc tggaaatatt tctgcgctgg ctgatgcgga aaatatttca
     5461 cgtaagatta ttacccgctg tatcaacacc gccaaattgc ctaaatcagt tgttgctctt
     5521 ttttctcacc ccggtgaact atctgcccgg tcaggtgatg cacttcaaaa agcctttaca
     5581 gataaagagg aattacttaa gcagcaggca tctaaccttc atgagcagaa aaaagctggg
     5641 gtgatatttg aagctgaaga agttatcact cttttaactt ctgtgcttaa aacgtcatct
     5701 gcatcaagaa ctagtttaag ctcacgacat cagtttgctc ctggagcgac agtattgtat
     5761 aagggcgata aaatggtgct taacctggac aggtctcgtg ttccaactga gtgtatagag
     5821 aaaattgagg ccattcttaa ggaacttgaa aagccagcac cctgatgcga cctcgtttta
     5881 gtctacgttt atctgtcttt acttaatgtc ctttgttaca ggccagaaag cataactggc
     5941 ctgaatattc tctctgggcc cactgttcca cttgtatcgt cggtctgata atcagactgg
     6001 gaccacggtc ccactcgtat cgtcggtctg attattagtc tgggaccacg gtcccactcg
     6061 tatcgtcggt ctgattatta gtctgggacc acggtcccac tcgtatcgtc ggtctgataa
     6121 tcagactggg accacggtcc cactcgtatc gtcggtctga ttattagtct gggaccatgg
     6181 tcccactcgt atcgtcggtc tgattattag tctgggacca cggtcccact cgtatcgtcg
     6241 gtctgattat tagtctggaa ccacggtccc actcgtatcg tcggtctgat tattagtctg
     6301 ggaccacggt cccactcgta tcgtcggtct gattattagt ctgggaccac gatcccactc
     6361 gtgttgtcgg tctgattatc ggtctgggac cacggtccca cttgtattgt cgatcagact
     6421 atcagcgtga gactacgatt ccatcaatgc ctgtcaaggg caagtattga catgtcgtcg
     6481 taacctgtag aacggagtaa cctcggtgtg cggttgtatg cctgctgtgg attgctgctg
     6541 tgtcctgctt atccacaaca ttttgcgcac ggttatgtgg acaaaatacc tggttaccca
     6601 ggccgtgccg gcacgttaac cgggctgcat ccgatgcaag tgtgtcgctg tcgacgagct
     6661 cgcgagctcg gacatgaggt tgccccgtat tcagtgtcgc tgatttgtat tgtctgaagt
     6721 tgtttttacg ttaagttgat gcagatcaat taatacgata cctgcgtcat aattgattat
     6781 ttgacgtggt ttgatggcct ccacgcacgt tgtgatatgt agatgataat cattatcact
     6841 ttacgggtcc tttccggtga tccgacaggt tacggggcgg cgacctcgcg ggttttcgct
     6901 atttatgaaa attttccggt ttaaggcgtt tccgttcttc ttcgtcataa cttaatgttt
     6961 ttatttaaaa taccctctga aaagaaagga aacgacaggt gctgaaagcg agctttttgg
     7021 cctctgtcgt ttcctttctc tgtttttgtc cgtggaatga acaatggaag tccgagctca
     7081 tcgctaataa cttcgtatag catacattat acgaagttat attcgatgcg gccgcaaggg
     7141 gttcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     7201 gattgtactg agagtgcacc atatgcggtg tgaaatacca cacagatgcg taaggagaaa
     7261 ataccgcatc aggcgccatt cgccattcag ctgcgcaact gttgggaagg gcgatcggtg
     7321 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     7381 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattgtaa
     7441 tacgactcac tatagggcga attcgagctc ggtacccggg gatcctctag agtcgacctg
     7501 caggcatgca agcttgagta ttctatagtc tcacctaaat agcttggcgt aatcatggtc
     7561 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     7621 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     7681 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     7741 ccaacgcgaa ccccttgcgg ccgcccgggc cgtcgaccaa ttctcatgtt tgacagctta
     7801 tcatcgaatt tctgccattc atccgcttat tatcacttat tcaggcgtag caaccaggcg
     7861 tttaagggca ccaataactg ccttaaaaaa attacgcccc gccctgccac tcatcgcagt
     7921 actgttgtaa ttcattaagc attctgccga catggaagcc atcacaaacg gcatgatgaa
     7981 cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt ataatatttg cccatggtga
     8041 aaacgggggc gaagaagttg tccatattgg ccacgtttaa atcaaaactg gtgaaactca
     8101 cccagggatt ggctgagacg aaaaacatat tctcaataaa ccctttaggg aaataggcca
     8161 ggttttcacc gtaacacgcc acatcttgcg aatatatgtg tagaaactgc cggaaatcgt
     8221 cgtggtattc actccagagc gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac
     8281 aagggtgaac actatcccat atcaccagct caccgtcttt cattgccata cgaaattccg
     8341 gatgagcatt catcaggcgg gcaagaatgt gaataaaggc cggataaaac ttgtgcttat
     8401 ttttctttac ggtctttaaa aaggccgtaa tatccagctg aacggtctgg ttataggtac
     8461 attgagcaac tgactgaaat gcctcaaaat gttctttacg atgccattgg gatatatcaa
     8521 cggtggtata tccagtgatt tttttctcca ttttagcttc cttagctcct gaaaatctcg
     8581 ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc
     8641 ttacgtgccg atcaacgtct cattttcgcc aaaagttggc ccagggcttc ccggtatcaa
     8701 cagggacacc aggatttatt tattctgcga agtgatcttc cgtcacaggt atttattcgc
     8761 gataagctca tggagcggcg taaccgtcgc acaggaagga cagagaaagc gcggatctgg
     8821 gaagtgacgg acagaacggt caggacctgg attggggagg cggttgccgc cgctgctgct
     8881 gacggtgtga cgttctctgt tccggtcaca ccacatacgt tccgccattc ctatgcgatg
     8941 cacatgctgt atgccggtat accgctgaaa gttctgcaaa gcctgatggg acataagtcc
     9001 atcagttcaa cggaagtcta cacgaaggtt tttgcgctgg atgtggctgc ccggcaccgg
     9061 gtgcagtttg tgatgccgga gtctgatgcg gttgcgatgc tgaaacaatt atcctgagaa
     9121 taaatgcctt ggcctttata tggaaatgtg gaactgagtg gatatgctgt ttttgtctgt
     9181 taaacagaga agctggctgt tatccactga gaagcgaacg aaacagtcgg gaaaatctcc
     9241 cattatcgta gagatccgca ttattaatct caggagcctg tgtagcgttt ataggaagta
     9301 gtgttctgtc atgatgcctg caagcggtaa cgaaaacgat ttgaatatgc cttcaggaac
     9361 aatagaaatc ttcgtgcggt gttacgttga agtggagcgg attatgtcag caatggacag
     9421 aacaacctaa tgaacacaga accatgatgt ggtctgtcct tttacagcca gtagtgctcg
     9481 ccgcagtcga gcgacagggc gaagcccatc gatactagct tgattgggat atctcgctca
     9541 tgtttgtcgc gtgctatgtc tttttaggta ctttgaacct acgttcgtac ttgtataata
     9601 tgatcatcgt attatcgttt ttcatccgtc cagcgcaaaa tgcattagca gctagtccta
     9661 gcgtgcggag ctacctggac aggtgcatga cggatgcgtg tccttcagtg actttctaat
     9721 taacagtaac ttctttactt atgtttcagt ttgtaagaag cgggattcgc tcgtcgcttg
     9781 acatctgatt ggactgcgtc ggcacgtgaa aactacattg tgaaatctgc taaaactccg
     9841 ggtatctctg acacaaaacg attcggcttc gcaatttcaa cattacggtc aaggctaacg
     9901 tatctttctc ggtcaacttc agattatgcc gattaaattg tcgtagcttt caaggcgttt
     9961 tgagtactgc ggcagttgtt gaacctgcaa ggagaagatc tcgacaacag aataaagcga
    10021 aaaatgggtc tcatgcacta acactcaggc ctccctcata atctctgttt gagtttacca
    10081 acaacacata tatacatttc gacaaaatga ccactcttga cgacacggct taccggtacc
    10141 gcaccagtgt cccgggggac gccgaggcca tcgaggcact ggatgggtcc ttcaccaccg
    10201 acaccgtctt ccgcgtcacc gccaccgggg acggcttcac cctgcgggag gtgccggtgg
    10261 acccgcccct gaccaaggtg ttccccgacg acgaatcgga cgacgaatcg gacgacgggg
    10321 aggacggcga cccggactcc cggacgttcg tcgcgtacgg ggacgacggc gacctggcgg
    10381 gcttcgtggt cgtctcgtac tccggctgga accgccggct gaccgtcgag gacatcgagg
    10441 tcgccccgga gcaccggggg cacggggtcg ggcgcgcgtt gatggggctc gcgacggagt
    10501 tcgcccgcga gcggggcgcc gggcacctct ggctggaggt caccaacgtc aacgcaccgg
    10561 cgatccacgc gtaccggcgg atggggttca ccctctgcgg cctggacacc gccctgtacg
    10621 acggcaccgc ctcggacggc gagcaggcgc tctacatgag catgccctgc ccctgaccga
    10681 cgccgaccaa caccgccggt ccgacgcggc ccgacgggtc cgaggcctcg gagatctggg
    10741 cccatgcggc cgcaacaact acctcgactt tggctgggac actttcagtg aggacaagaa
    10801 gcttcagaag cgtgctatcg aactcaacca gggacgtgcg gcacaaatgg gcatccttgc
    10861 tctcatggtg cacgaacagt tgggagtctc tatccttcct taaaaattta attttcatta
    10921 gttgcagtca ctccgctttg gtttcgtaac tataacggtc ctaaggtagc gaacgttgca
    10981 ggccatgctg tccaggcagg tagatgacga ccatcaggga cagcttcaag gatcgctcgc
    11041 ggctcttacc agcctaactt cgatcattgg accgctgatc gtcacggcga tttatgccgc
    11101 ctcggcgagc acatggaacg ggttggcatg gattgtaggc gccgccctat accttgtctg
    11161 cctccccgcg ttgcgtcgcg gtgcatggag ccgggccacc tcgacctgaa tggaagccgg
    11221 cggcacctcg ctaacggatt caccactcca agaattggag ccaatcaatt cttgcggaga
    11281 actgtgaatg cgcaaaccaa cccttggcag aacatatcca tcgcgtccgc catctccagc
    11341 agccgcacgc ggcgcatcgg gggggggggg gtttcaattc atcatttttt ttttattctt
    11401 ttttttgatt tcggtttcct tgaaattttt ttgattcggt aatctccgaa cagaaggaag
    11461 aacgaaggaa ggagcacaga cttagattgg tatatatacg catatgtagt gttgaagaaa
    11521 catgaaattg cccagtattc ttaacccaac tgcacagaac aaaaacctgc aggaaacgaa
    11581 gataaatcat gtcgaaagct acatataagg aacgtgctgc tactcatcct agtcctgttg
    11641 ctgccaagct atttaatatc atgcacgaaa agcaaacaaa cttgtgtgct tcattggatg
    11701 ttcgtaccac caaggaatta ctggagttag ttgaagcatt aggtcccaaa atttgtttac
    11761 taaaaacaca tgtggatatc ttgactgatt tttccatgga gggcacagtt aagccgctaa
    11821 aggcattatc cgccaagtac aattttttac tcttcgaaga cagaaaattt gctgacattg
    11881 gtaatacagt caaattgcag tactctgcgg gtgtatacag aatagcagaa tgggcagaca
    11941 ttacgaatgc acacggtgtg gtgggcccag gtattgttag cggtttgaag caggcggcgg
    12001 aagaagtaac aaaggaacct agaggccttt tgatgttagc agaattgtca tgcaagggct
    12061 ccctagctac tggagaatat actaagggta ctgttgacat tgcgaagagc gacaaagatt
    12121 ttgttatcgg ctttattgct caaagagaca tgggtggaag agatgaaggt tacgattggt
    12181 tgattatgac acccggtgtg ggtttagatg acaagggaga cgcattgggt caacagtata
    12241 gaaccgtgga tgatgtggtc tctacaggat ctgacattat tattgttgga agaggactat
    12301 ttgcaaaggg aagggatgct aaggtagagg gtgaacgtta cagaaaagca ggctgggaag
    12361 catatttgag aagatgcggc cagcaaaact aaaaaactgt attataagta aatgcatgta
    12421 tactaaactc acaaattaga gcttcaattt aattatatca gttattaccc ggccgggaat
    12481 ctcggtcgta atgattttta taatgacgaa aaaaaaaaaa ttggaaagaa aacccccccc
    12541 cccccgcagc gttgggtcct ggccacgggt gcgcatgatc gtgctcctgt cgttgaggac
    12601 ccggctaggc tggcggggtt gccttactgg ttagcagaat gaatcaccga tacgcgagcg
    12661 aacgtgaagc gactgctgct gcaaaacgtc tgcgacctga gcaacaacat gaatggtctt
    12721 cggtttccgt gtttcgtaaa gtctggaaac gcggaagtca gcgccctgca ccattatgtt
    12781 ccggatctgc atcgcaggat gctgctggct accctgtgga acacctacat ctgtattaac
    12841 gaagcgctgg cattgaccct gagtgatttt tctctggtcc cgccgcatcc ataccgccag
    12901 ttgtttaccc tcacaacgtt ccagtaaccg ggcatgttca tcatcagtaa cccgtatcgt
    12961 gagcatcctc tctcgtttca tcgagttacg ctagggataa cagggtaata tagatcttcc
    13021 gctgcataac cctgcttcgg ggtcattata gcgatttttt cggtatatcc atcctttttc
    13081 gcacgatata caggattttg ccaaagggtt cgtgtagact ttccttggtg tatccaacgg
    13141 cgtcagccgg gcaggatagg tgaagtaggc ccacccgcga gcgggtgttc cttcttcact
    13201 gtcccttatt cgcacctggc ggtgctcaac gggaatcctg ctctgcgagg ctggccggct
    13261 accgccggcg taacagatga gggcaagcgg atggctgatg aaaccaagcc aaccaggaag
    13321 ggcagcccac ctatcaaggt gtactgcctt ccagacgaac gaagagcgat tgaggaaaag
    13381 gcggcggcgg ccggcatgag cctgtcggcc tacctgctgg ccgtcggcca gggctacaaa
    13441 atcacgggcg tcgtggacta tgagcacgtc cgcgagctgg cccgcatcaa tggcgacctg
    13501 ggccgcctgg gcggcctgct gaaactctgg ctcaccgacg acccgcgcac ggcgcggttc
    13561 ggtgatgcca cgatcctcgc cctgctggcg aagatcgaag agaagcagga cgagcttggc
    13621 aaggtcatga tgggcgtggt ccgcccgagg gcagagccat gactttttta gccgctaaaa
    13681 cggccggggg gtgcgcgtga ttgccaagca cgtccccatg cgctccatca agaagaggca
    13741 cttcgagctg taagtacatc accgacgagc aaggcaagac gatccgcgcc tgttttattg
    13801 agaacgttgt tcgtgttggc ctcaatggta gcgatgcgtc attcagcgaa gttctggtgc
    13861 tgatgatgtg gttcgctttg ccactggtca atgtggtaag cccgtgtaat gtcagtaacc
    13921 tttttactga tctcagcttg agcacggtcg ctgatgagct tatccatggc cccacggtaa
    13981 cggatatgat cctctagggc gttgacaaat tctttgtcgg ttttcatggg gtagacgtca
    14041 gtgaccaggg aacgtctccc tacaaaaagt tgcgcatact ttgctccgtt gtcgacggca
    14101 ggggtgtcgg accagattgt atctgtggca acggcctcat tgcgtcaatg gacctttaaa
    14161 gccgggaaac gagacttgaa atgtttacgt agaggtgcat tatagacctc acgcgcatat
    14221 tgagtggttg caaaaatggt tttgcgaacg gtatcactgg agacccatgc caggcaagga
    14281 cggagggcat cataatcatg ttcattgcgt tggacagcat gcttgttaca cgtaagtata
    14341 cggtcgagag catcgtgacg ttgagaatta cagatggcaa cgtgtcggtg catgtattgg
    14401 ccgagattgt caaatcgtgg ctcaatatat ggcatgccag gtaagtcatg gatgtcggta
    14461 tgccattcag ccttcatgtc gatttcgttg tcgagaactg atgggtccca gtcaacgtca
    14521 gaagtaagaa caacatgggg aagggaatga agttcatgac tggtgggtgc gcgcatatcc
    14581 atgtacggaa gaccctgtcg tatgtttagt gggatgatgt aatcatccag ggtaacaata
    14641 cgttggttgc cgcccacggt acgagaccga tcttgaacac aattgtggca atgttcaagt
    14701 tgggcgctgg aatggatggt tttaccttta ccaagatgcg cgtattgatg catgataaca
    14761 acaatgggcc ctcgttgcga ttcgacaaga ccggcagccg tgacaatgtc tagattggac
    14821 aaggtatgtt catttatacc ggttatgttg gctgaacgtc cagttttgtg cagtacggta
    14881 acatcactcc ccgcaagtcc accgttggca ccgcgatcga caagtgcaga tgtggtattt
    14941 tgtactgtgt gtcgggagac ttggtactgt aacacatttg actgaagtga cttggtggta
    15001 ccatgaggac ggagggaacc cgttgatacg gtgttgcgtg acgctgcaag taccttacgt
    15061 atgtcgcctg gatccatacg tccaacccga tcagaaagat gtgctaggag ttccgtttct
    15121 ggtgcacaat catgaaaggt atccacggga gttccattgt tgtcgtccgg ggctgcgtcg
    15181 cgactgccag ttgcatgtac atgcgccatc aagggacgtg ccaatccatt actaaccggg
    15241 agaaccttag ctggggcagc caatccctga agtatagcct tagcatcatc tgagagctgg
    15301 ttccacatat ctctagggat ataaggtcgt tcacggttag tgggcgtact atgatgtcgg
    15361 ttattagtac aattctccct gcgggcatgc gcattggctt catagagtac ggaaggtgac
    15421 aagtcaatat tgtagttaac atccggatca gtgtcaaagt ctgtaggatg gtaagaaaga
    15481 tcagtagaat gaatactacg cttttccttg ggactacggg aattagagaa gttgtttcct
    15541 ttattgtaga gtgacgctga agcaagtaga agactaaggt aactctcgta gctaatagga
    15601 ttacctcctt tggctaggtc aagagtggct gtgatcttca cttgacaaag ttccggtaca
    15661 ttgtggacag cattctccaa aagactaaga cacagttgct ttgggagttg ctcagccatt
    15721 ggtacagtat tgtggtagat acaaaggtgg ttcttccaat gaaggataaa tccttctgct
    15781 gtgcctgtcc atgaggatcc atatttcgcc gtagttaggt aaccaagcgt ggtggctgaa
    15841 ctgatcttcg cacttgctga ttccgtatag tgtttgacaa ctttacaaaa cacttcttgc
    15901 gcagttcgct ctagcctgga cagagaaggg acaatcttcg tcgttaagac tcgtgcgaat
    15961 agcaaaagat cacaaaatag cacatcggca ccgaccaacg attatttcca aggaaaaaaa
    16021 gaatgcttca ctacaagaaa ttgtgtcatc cctatacaga gtcttgttac tgtgacagaa
    16081 aattgatgga agatgtggcg gattgccttt acactagcca acttgttcga ctaattgcag
    16141 cttcttctga gaggcttcac cgagtaacgc gaagaacacc ggtgtctcgt acatgctcgt
    16201 cggtgaacgc tcgtccaatg acacccccca ctttgtatca atatcccaac ttggtagtga
    16261 actggaatga tacatgcaat ttcgcgccgc atcaacagcc acgggccacc atcgacgaat
    16321 agactcgg
//
