LOCUS       sequence-452520-       10970 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Release plasmid with Cas9 working cassette, apramycin resistance, 
            and pSG5 replicon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10970)
  AUTHORS   Xie F, Zhao H, Liu J, Yang X, Neuber M, Agrawal AA, Kaur A, Herrmann
            J, Kalinina OV, Wei X, Muller R, Fu C
  TITLE     Autologous DNA mobilization and multiplication expedite natural 
            products discovery from bacteria.
  JOURNAL   Science. 2024 Dec 13;386(6727):eabq7333. doi: 
            10.1126/science.abq7333. Epub 2024 Dec 13.
  PUBMED    39666857
REFERENCE   2  (bases 1 to 10970)
  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: "Science."; 
            date: "2024-12-13"; volume: "386(6727):eabq7333. doi"; pages: " 
            10.1126/science.abq7333. Epub 2024 Dec 13"
COMMENT     Sequence Label: pRel
FEATURES             Location/Qualifiers
     source          1..10970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             423..1865
                     /codon_start=1
                     /gene="rep"
                     /product="replication initiator protein from the 
                     Streptomyces ghanaensis plasmid pSG5"
                     /label=pSG5 Rep
                     /translation="MWKPGEATWGNTCRCNNVHTCPWCMSRILAVRGSNVQLAADGLAD
                     AGYGLHLGTNTLRHFERMAFGTVRKGMRHGLVAVLHDGWKGAYGSSGRRWRTMRDDFGI
                     IGYERAFEDTFGWGSGWHLHWHTLWVTREVLGPDAQAAFRDALAGAWAAGVESAGGYTV
                     SETCDRPGCSCEGKGHGTDVRPLNGADAADGDAGKQARYLYKDGDKTKGGVAKIGLELA
                     GQNFKAGRGDDRMGPLDLGDAAAAELQRLRRPGPFVEKYREREFGVFQVRKHYRSQNLN
                     RLIKELGIQQDVRTEEEITDDTEGLVAIAVIPAYIWYRYIARVAGRRLDLIKVAETYGL
                     PGVRRLVESWGLVWGKDVLDPPAPEAPAAPGDLDADQMRFEVMSEEEAAFREARRKANE
                     ARTEELAASLDRVRQPKKEAIRPTISLRKRLKPKPVTVDVKTPPPGAASPVCRRCKGKL
                     APVLQPWGQHPGDCLRVDTAVA"
     oriT            2527..2636
                     /note="incP origin of transfer"
     CDS             2952..3752
                     /codon_start=1
                     /gene="aac(3)IV"
                     /product="aminoglycoside 3-N-acetyltransferase type IV     
                               "
                     /label=ApmR
                     /note="confers resistance to apramycin (Paget and Davies, 
                     1996)"
                     /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
                     LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDSFWRLPNV
                     KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
                     LAELLAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     rep_origin      4054..4641
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4542..4561
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4795..4812
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4929..4950
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4965..4970,4971..4988,4989..4995)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     misc_RNA        5203..5278
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        5497..5601
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5602..5670,5671..6462)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5820..5839)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             6728..10834
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             6728..10831
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
ORIGIN
        1 tttgctctgt gacgagcgta cgagagaaca gcgcgtcgac accaccgcgc cccgccggac
       61 cgaagtgcca ctgcggcacc ccgatcgtcc acatccccgg gaagcgcccc aaaatctact
      121 gctcggacgc ctgccgaaag cgcgcaaagc gtctcattgc ggaccagtca cgcagcgtag
      181 caagcacgac tacagcgggt aatccggacg cgggctttag tagtaaggga aaatctccgt
      241 ccactgcagg tcagagcgct gtaactggaa cgggttctac cccaaacgag ggtgccgcga
      301 cagccgggcg ggaggacccg cgcgacgagc ggttcgcccg gcgggaccgg caccagacgg
      361 tgtcgctgaa cgaggcgttc aagggctgcg ggacccggct cactgccggg acggccgagc
      421 tgatgtggaa gccgggcgag gcgacgtggg gcaacacgtg ccgctgcaac aacgtccaca
      481 cctgcccgtg gtgcatgagc cgcatcctgg ccgtacgggg ctcgaacgtg cagctcgccg
      541 ccgacggtct cgcggacgcc gggtacgggc tgcaccttgg cacgaacacg ttgcgccact
      601 tcgagcgcat ggcgttcggc accgtccgca agggaatgcg gcacggcctg gtcgcggtct
      661 tgcacgacgg ctggaagggc gcgtacggct cgtccggccg ccgctggcgc acgatgcgcg
      721 acgacttcgg gatcatcggc tacgagcggg ccttcgagga caccttcggc tggggctccg
      781 gctggcacct gcactggcac acgctctggg tcacccgcga ggtgctcggc ccggacgccc
      841 aggccgcgtt ccgcgacgca ctcgccggag cctgggccgc cggggtcgaa tccgcgggcg
      901 gctacaccgt cagcgagacg tgcgaccggc ccggttgctc ctgtgagggc aagggccacg
      961 gcaccgacgt gcgcccactc aacggcgcgg acgccgctga cggcgatgcg ggcaagcagg
     1021 cccgctacct ctacaaggac ggcgacaaga ccaagggcgg cgtcgccaag atcggcctgg
     1081 agctggccgg acagaacttc aaggccggtc gcggcgacga ccgcatggga ccgctcgacc
     1141 tgggcgacgc ggcggccgcc gagctgcagc ggcttcgtcg gcccggcccg ttcgtcgaga
     1201 agtaccgcga gcgcgagttc ggcgtcttcc aggtccgcaa gcactaccgg tcgcagaacc
     1261 tgaaccggct gatcaaggaa ctcggcatcc agcaggacgt ccgcaccgag gaggagatca
     1321 ccgacgacac cgagggcctg gtcgccatcg cggtcatccc ggcctacatc tggtaccgat
     1381 acatcgcccg cgtcgccgga cgtcgcctcg acctcatcaa ggtcgccgag acgtacggcc
     1441 taccgggcgt ccggcggctc gtcgagtcct ggggcctggt gtggggcaag gacgtcctgg
     1501 acccgcccgc cccggaagcg ccggccgcgc cgggcgacct tgacgctgac cagatgaggt
     1561 tcgaggtcat gtccgaggag gaagctgctt tccgcgaggc gcgccggaag gccaacgagg
     1621 cccgcacgga ggagctggcc gcttcgctcg accgagtgcg tcagccgaag aaggaggcga
     1681 tccgcccgac gatcagcctt cggaagcgcc tgaagccgaa gccggtcacc gtcgacgtca
     1741 agactccccc gcccggcgcc gcgtcgcccg tgtgccgccg ctgcaagggc aagctcgccc
     1801 cggtactcca gccctggggc cagcaccccg gagactgcct ccgcgtcgac acagcggtcg
     1861 cgtagggccg ctgagggacc gccatgagac cgccaccccg gacatccggg tggcggtctc
     1921 atggcggtcc cggcgcgaaa gccgagggtc tgcctgccgt gaggtggccg gcggacctgg
     1981 gttactgctg gaatcccgcg cccagtccga gcgtccgccg gcgcttttca cctggctccg
     2041 ggtgctcgtc gatgagcacc aggccgtcga cgacgacgcg tccgcctgcc tgcttttggc
     2101 cggcgggccg ggccgcggtt gtccgccggg tcggaccggt cccttggtcg accgatgcct
     2161 cggccacgac gatgctgggc ccgaagcggt acgcgacccg tgccgggccg cagcggatcg
     2221 cggcagcagg agctccccgc ccgccacggc tggctggatg gtcgcctcac gcgcccgcag
     2281 gggtgacggc gtgggtgcgg cggtcagccc cgtgatcggg cgtcagacgg ccgctgaggg
     2341 accgcgacca cccctccctt cccgcctgcc gtctttcacc gcccggccgg tccgcagacg
     2401 ccgtacggcc aggccacgga ggattacggg cagatggcgc ccacccggca cccgggctgg
     2461 ggcaccctgg acagcacgaa ggtccgcccc actcaccggg gcgggccttc gcgttgagct
     2521 agagttcccg ccagcctcgc agagcaggat tcccgttgag caccgccagg tgcgaataag
     2581 ggacagtgaa gaaggaacac ccgctcgcgg gtgggcctac ttcacctatc ctgcccggct
     2641 gacgccgttg gatacaccaa ggaaagtcta cacgaaccct ttggcaaaat cctgtatatc
     2701 gtgcgaaaaa ggatggatat accgaaaaaa tcgctataat gaccccgaag cagggttatg
     2761 cagcggaaaa tgcagctcac ggtaactgat gccgtatttg cagtaccagc gtacggccca
     2821 cagaatgatg tcacgctgaa aatgccggcc tttgaatggg ttcatgtgca gctccataag
     2881 caaaagggga tgataagttt atcaccaccg actatttgca acagtgccgt tgatcgtgct
     2941 atgatcgact gatgtcatca gcggtggagt gcaatgtcgt gcaatacgaa tggcgaaaag
     3001 ccgagctcat cggtcagctt ctcaaccttg gggttacccc cggcggtgtg ctgctggtcc
     3061 acagctcctt ccgtagcgtc cggcccctcg aagatgggcc acttggactg atcgaggccc
     3121 tgcgtgctgc gctgggtccg ggagggacgc tcgtcatgcc ctcgtggtca ggtctggacg
     3181 acgagccgtt cgatcctgcc acgtcgcccg ttacaccgga ccttggagtt gtctctgact
     3241 cattctggcg cctgccaaat gtaaagcgca gcgcccatcc atttgccttt gcggcagcgg
     3301 ggccacaggc agagcagatc atctctgatc cattgcccct gccacctcac tcgcctgcaa
     3361 gcccggtcgc ccgtgtccat gaactcgatg ggcaggtact tctcctcggc gtgggacacg
     3421 atgccaacac gacgctgcat cttgccgagt tgttggcaaa ggttccctat ggggtgccga
     3481 gacactgcac cattcttcag gatggcaagt tggtacgcgt cgattatctc gagaatgacc
     3541 actgctgtga gcgctttgcc ttggcggaca ggtggctcaa ggagaagagc cttcagaagg
     3601 aaggtccagt cggtcatgcc tttgctcggt tgatccgctc ccgcgacatt gtggcgacag
     3661 ccctgggtca actgggccga gatccgttga tcttcctgca tccgccagag gcgggatgcg
     3721 aagaatgcga tgccgctcgc cagtcgattg gctgagctca taagttccta ttccgaagtt
     3781 cctattcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     3841 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     3901 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaaaagga tctaggtgaa
     3961 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4021 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4081 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4141 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4201 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4261 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4321 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4381 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     4441 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     4501 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     4561 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     4621 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     4681 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     4741 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     4801 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     4861 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     4921 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     4981 ccggctcgta tgttgtgtgg aattgtgagt ttaaacgcgg ataacaattt cacacagtcg
     5041 acttctagtg cagctagtgt cgactagagg atccccgggt atgcggtcga tcttgacggc
     5101 tggcgagagg tgcggggagg atctgaccga cgcggtccac acgtggcacc gcgatgctgt
     5161 tgtgggcaca atcgtgccgg ttggtaggat cgacggccat gggttttagg tctagaaata
     5221 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
     5281 tttttacgta ggcgtcagct acgatgttcc ggggactgct gatccggtca gcaggtggaa
     5341 gagggactgg attccaaagt tctcaatgct gcttgctgtt cttgaatggg gggtcgttga
     5401 cgacgacatg gctcgattgg cgcgacaagt tgctgcgatt ctcaccaata aaaaacgccc
     5461 ggcggcagaa caaatccaga tggagttcac tagttgcgcg gaacccctat ttgtttattt
     5521 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5581 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     5641 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     5701 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     5761 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     5821 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     5881 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     5941 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     6001 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     6061 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     6121 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     6181 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     6241 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     6301 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     6361 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     6421 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     6481 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6541 atcctttttg ataatctcat gaccaaaatg ctagctgagg tcattactgg accggatcgg
     6601 ggatctgggc tgagggagcc gacggcacgc ggcggctcac ggcgtggcac gcggaacgtc
     6661 cgggcttgca cctcacgtca cgtgaggagg cagcgtggac ggcgtcagag aagggagcgg
     6721 acaattgatg gacaagaagt actccatcgg cctggacatc ggcaccaact ccgtcggctg
     6781 ggccgtcatc accgacgagt acaaggtccc gtccaagaag ttcaaggtcc tgggcaacac
     6841 cgaccgccac tccatcaaga agaacctgat cggcgccctg ctgttcgact ccggcgagac
     6901 cgccgaggcc acccgcctga agcgcaccgc ccgccgccgc tacacccgcc gcaagaaccg
     6961 catctgctac ctgcaggaga tcttctccaa cgagatggcc aaggtcgacg actccttctt
     7021 ccaccgcctg gaggagtcct tcctggtcga ggaggacaag aagcacgagc gccacccgat
     7081 cttcggcaac atcgtcgacg aggtcgccta ccacgagaag tacccgacca tctaccacct
     7141 gcgcaagaag ctggtcgact ccaccgacaa ggccgacctg cgcctgatct acctggccct
     7201 ggcccacatg atcaagttcc gcggccactt cctgatcgag ggcgacctga acccggacaa
     7261 ctccgacgtc gacaagctgt tcatccagct ggtccagacc tacaaccagc tgttcgagga
     7321 gaacccgatc aacgcctccg gcgtcgacgc caaggccatc ctgtccgccc gcctgtccaa
     7381 gtcccgccgc ctggagaacc tgatcgccca gctgccgggc gagaagaaga acggcctgtt
     7441 cggcaacctg atcgccctgt ccctgggcct gaccccgaac ttcaagtcca acttcgacct
     7501 ggccgaggac gccaagctgc agctgtccaa ggacacctac gacgacgacc tggacaacct
     7561 gctggcccag atcggcgacc agtacgccga cctgttcctg gccgccaaga acctgtccga
     7621 cgccatcctg ctgtccgaca tcctgcgcgt caacaccgag atcaccaagg ccccgctgtc
     7681 cgcctccatg atcaagcgct acgacgagca ccaccaggac ctgaccctgc tgaaggccct
     7741 ggtccgccag cagctgccgg agaagtacaa ggagatcttc ttcgaccagt ccaagaacgg
     7801 ctacgccggc tacatcgacg gcggcgcctc ccaggaggag ttctacaagt tcatcaagcc
     7861 gatcctggag aagatggacg gcaccgagga gctgctggtc aagctgaacc gcgaggacct
     7921 gctgcgcaag cagcgcacct tcgacaacgg ctccatcccg caccagatcc acctgggcga
     7981 gctgcacgcc atcctgcgcc gccaggagga cttctacccg ttcctgaagg acaaccgcga
     8041 gaagatcgag aagatcctga ccttccgcat cccgtactac gtcggcccgc tggcccgcgg
     8101 caactcccgc ttcgcctgga tgacccgcaa gtccgaggag accatcaccc cgtggaactt
     8161 cgaggaggtc gtcgacaagg gcgcctccgc ccagtccttc atcgagcgca tgaccaactt
     8221 cgacaagaac ctgccgaacg agaaggtcct gccgaagcac tccctgctgt acgagtactt
     8281 caccgtctac aacgagctga ccaaggtcaa gtacgtcacc gagggcatgc gcaagccggc
     8341 cttcctgtcc ggcgagcaga agaaggccat cgtcgacctg ctgttcaaga ccaaccgcaa
     8401 ggtcaccgtc aagcagctga aggaggacta cttcaagaag atcgagtgct tcgactccgt
     8461 cgagatctcc ggcgtcgagg accgcttcaa cgcctccctg ggcacctacc acgacctgct
     8521 gaagatcatc aaggacaagg acttcctgga caacgaggag aacgaggaca tcctggagga
     8581 catcgtcctg accctgaccc tgttcgagga ccgcgagatg atcgaggagc gcctgaagac
     8641 ctacgcccac ctgttcgacg acaaggtcat gaagcagctg aagcgccgcc gctacaccgg
     8701 ctggggccgc ctgtcccgca agctgatcaa cggcatccgc gacaagcagt ccggcaagac
     8761 catcctggac ttcctgaagt ccgacggctt cgccaaccgc aacttcatgc agctgatcca
     8821 cgacgactcc ctgaccttca aggaggacat ccagaaggcc caggtctccg gccagggcga
     8881 ctccctgcac gagcacatcg ccaacctggc cggctccccg gccatcaaga agggcatcct
     8941 gcagaccgtc aaggtcgtcg acgagctggt caaggtcatg ggccgccaca agccggagaa
     9001 catcgtcatc gagatggccc gcgagaacca gaccacccag aagggccaga agaactcccg
     9061 cgagcgcatg aagcgcatcg aggagggcat caaggagctg ggctcccaga tcctgaagga
     9121 gcacccggtc gagaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa
     9181 cggccgcgac atgtacgtcg accaggagct ggacatcaac cgcctgtccg actacgacgt
     9241 cgaccacatc gtcccgcagt ccttcctgaa ggacgactcc atcgacaaca aggtcctgac
     9301 ccgctccgac aagaaccgcg gcaagtccga caacgtcccg tccgaggagg tcgtcaagaa
     9361 gatgaagaac tactggcgcc agctgctgaa cgccaagctg atcacccagc gcaagttcga
     9421 caacctgacc aaggccgagc gcggcggcct gtccgagctg gacaaggccg gcttcatcaa
     9481 gcgccagctg gtcgagaccc gccagatcac caagcacgtc gcccagatcc tggactcccg
     9541 catgaacacc aagtacgacg agaacgacaa gctgatccgc gaggtcaagg tcatcaccct
     9601 gaagtccaag ctggtctccg acttccgcaa ggacttccag ttctacaagg tccgcgagat
     9661 caacaactac caccacgccc acgacgccta cctgaacgcc gtcgtcggca ccgccctgat
     9721 caagaagtac ccgaagctgg agtccgagtt cgtctacggc gactacaagg tctacgacgt
     9781 ccgcaagatg atcgccaagt ccgagcagga gatcggcaag gccaccgcca agtacttctt
     9841 ctactccaac atcatgaact tcttcaagac cgagatcacc ctggccaacg gcgagatccg
     9901 caagcgcccg ctgatcgaga ccaacggcga gaccggcgag atcgtctggg acaagggccg
     9961 cgacttcgcc accgtccgca aggtcctgtc catgccgcag gtcaacatcg tcaagaagac
    10021 cgaggtccag accggcggct tctccaagga gtccatcctg ccgaagcgca actccgacaa
    10081 gctgatcgcc cgcaagaagg actgggaccc gaagaagtac ggcggcttcg actccccgac
    10141 cgtcgcctac tccgtcctgg tcgtcgccaa ggtcgagaag ggcaagtcca agaagctgaa
    10201 gtccgtcaag gagctgctgg gcatcaccat catggagcgc tcctccttcg agaagaaccc
    10261 gatcgacttc ctggaggcca agggctacaa ggaggtcaag aaggacctga tcatcaagct
    10321 gccgaagtac tccctgttcg agctggagaa cggccgcaag cgcatgctgg cctccgccgg
    10381 cgagctgcag aagggcaacg agctggccct gccgtccaag tacgtcaact tcctgtacct
    10441 ggcctcccac tacgagaagc tgaagggctc cccggaggac aacgagcaga agcagctgtt
    10501 cgtcgagcag cacaagcact acctggacga gatcatcgag cagatctccg agttctccaa
    10561 gcgcgtcatc ctggccgacg ccaacctgga caaggtcctg tccgcctaca acaagcaccg
    10621 cgacaagccg atccgcgagc aggccgagaa catcatccac ctgttcaccc tgaccaacct
    10681 gggcgccccg gccgccttca agtacttcga caccaccatc gaccgcaagc gctacacctc
    10741 caccaaggag gtcctggacg ccaccctgat ccaccagtcc atcaccggcc tgtacgagac
    10801 ccgcatcgac ctgtcccagc tgggcggcga ctaaggatcg gacgaacaat aaggggagcg
    10861 ggaaaccgct cccctttttt attgataaca aaaaagcttc tcgtcgaagg cactagaagg
    10921 cctatatgcg gggatcgacc gcgcagcatg ccgagtgagg acgtttaaac
//
