LOCUS       sequence-237466-       13375 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  High efficiency Streptomyces genome editing by CRISPR/Cas9 system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13375)
  AUTHORS   Huang H, Zheng G, Jiang W, Hu H, Lu Y
  TITLE     One-step high-efficiency CRISPR/Cas9-mediated genome editing in 
            Streptomyces.
  JOURNAL   Acta Biochim Biophys Sin (Shanghai). 2015 Apr;47(4):231-43. doi: 
            10.1093/abbs/gmv007. Epub 2015 Mar 3.
  PUBMED    25739462
REFERENCE   2  (bases 1 to 13375)
  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"; doi: 
            "10.1093/abbs/gmv007"; journalName: "Acta Biochim Biophys Sin 
            (Shanghai)"; date: "2015-04"; volume: "47"; issue: "4"; pages: 
            "231-43"
COMMENT     Sequence Label: pKCcas9dO
FEATURES             Location/Qualifiers
     source          1..13375
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(547..1989)
                     /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            3156..3265
                     /note="incP origin of transfer"
     CDS             3298..3669
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     primer_bind     4355..4377
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4369..4386
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4370..4386
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_RNA        complement(6428..6503)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(6524..6558)
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     CDS             6770..10876
                     /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             6770..10873
                     /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"
     primer_bind     complement(10891..10907)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10891..10907)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10904..10926)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10915..10931
                     /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(10939..10945,10946..10963,10964..10969))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10984..11005
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11122..11139)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11293..11881)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11373..11392)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             12085..12885
                     /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
                     LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
                     KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
                     LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
ORIGIN
        1 tgtgaggagg ctggtccttt accctgatgc ccgagggcat cagggtaaag gaccagcctc
       61 gccaactagg agcgctacca aggcgaacac aacccaggac ttggtagaac ccttcgccgg
      121 agaagttcag actcatcggc atgagcaccc ctcgccgcgc actcggcacc ggtccgtcga
      181 ccaccaccag gacgtcgttg tcgacgtcgg ccccgcggct cctgcccgcc gaacgcgtcg
      241 tcgtcgacgg cctggtgctc atcgacgagc acccggagcc aggtgaaaag cgccggcgga
      301 cgctcggact gggcgcggga ttccagcagt aacccaggtc cgccggccac ctcacggcag
      361 gcagaccctc ggctttcgcg ccgggaccgc catgagaccg ccacccggat gtccggggtg
      421 gcggtctcat ggcggtccct cagcggccct acgcgaccgc tgtgtcgacg cggaggcagt
      481 ctccggggtg ctggccccag ggcyggakkw ccggggyggc ggtctcatgg cggtccctca
      541 gcggccctac gcgaccgctg tgtcgacgcg gaggcagtct ccggggtgct ggccccaggg
      601 ctggagtacc ggggcgagct tgcccttgca gcggcggcac acgggcgacg cggcgccggg
      661 cgggggagtc ttgacgtcga cggtgaccgg cttcggcttc aggcgcttcc gaaggctgat
      721 cgtcgggcgg atcgcctcct tcttcggctg acgcactcgg tcgagcgaag cggccagctc
      781 ctccgtgcgg gcctcgttgg ccttccggcg cgcctcgcgg aaagcagctt cctcctcgga
      841 catgacctcg aacctcatct ggtcagcgtc aaggtcgccc ggcgcggccg gcgcttccgg
      901 ggcgggcggg tccaggacgt ccttgcccca caccaggccc caggactcga cgagccgccg
      961 gacgcccggt aggccgtacg tctcggcgac cttgatgagg tcgaggcgac gtccggcgac
     1021 gcgggcgatg tatcggtacc agatgtaggc cgggatgacc gcgatggcga ccaggccctc
     1081 ggtgtcgtcg gtgatctcct cctcggtgcg gacgtcctgc tggatgccga gttccttgat
     1141 cagccggttc aggttctgcg accggtagtg cttgcggacc tggaagacgc cgaactcgcg
     1201 ctcgcggtac ttctcgacga acgggccggg ccgacgaagc cgctgcagct cggcggccgc
     1261 cgcgtcgccc aggtcgagcg gtcccatgcg gtcgtcgccg cgaccggcct tgaagttctg
     1321 tccggccagc tccaggccga tcttggcgac gccgcccttg gtcttgtcgc cgtccttgta
     1381 gaggtagcgg gcctgcttgc ccgcatcgcc gtcagcggcg tccgcgccgt tgagtgggcg
     1441 cacgtcggtg ccgtggccct tgccctcaca ggagcaaccg ggccggtcgc acgtctcgct
     1501 gacggtgtag ccgcccgcgg attcgacccc ggcggcccag gctccggcga gtgcgtcgcg
     1561 gaacgcggcc tgggcgtccg ggccgagcac ctcgcgggtg acccagagcg tgtgccagtg
     1621 caggtgccag ccggagcccc agccgaaggt gtcctcgaag gcccgctcgt agccgatgat
     1681 cccgaagtcg tcgcgcatcg tgcgccagcg gcggccggac gagccgtacg cgcccttcca
     1741 gccgtcgtgc aagaccgcga ccaggccgtg ccgcattccc ttgcggacgg tgccgaacgc
     1801 catgcgctcg aagtggcgca acgtgttcgt gccaaggtgc agcccgtacc cggcgtccgc
     1861 gagaccgtcg gcggcgagct gcacgttcga gccccgtacg gccaggatgc ggctcatgca
     1921 ccacgggcag gtgtggacgt tgttgcagcg gcacgtgttg ccccacgtcg cctcgcccgg
     1981 cttccacatc agctcggccg tcccggcagt gagccgggtc ccgcagccct tgaacgcctc
     2041 gttcagcgac accgtctggt gccggtcccg ccgggcgaac cgctcgtcgc gcgggtcctc
     2101 ccgcccggct gtcgcggcac cctcgtttgg ggtagaaccc gttccagtta cagcgctctg
     2161 acctgcagtg gacggagatt ttcccttact actaaagccc gcgtccggat tacccgctgt
     2221 agtcgtgctt gctacgctgc gtgactggtc cgcaatgaga cgctttgcgc gctttcggca
     2281 ggcgtccgag cagtagattt tggggcgctt cccggggatg tggacgatcg gggtgccgca
     2341 gtggcacttc ggtccggcgg ggcgcggtgg tgtcgacgcg ctgttctctc gtacgctcgt
     2401 cacagagcaa acgtcctcac tcggcatgct gcgccggttc gggggcggcg agcccgggag
     2461 gccaatcccg ggctcgtgcc atttctgggt cctgttgatc ctggcattgg tgtggccgtt
     2521 cattgcccct gctcgctcct gacgcgccga tagacgtccg atacgcccgg tgctggtggg
     2581 atttgatagg tcggaagaag ccccgcccgg gcctgggcgg ggcttcctgt gcgtcaggac
     2641 ctcctcgtcg tgagcctctt cggcctatgg acggagtgac ctcgtgatcc gttacagccg
     2701 cgcgcgctcg cgtagagcgg tctcatcagt tccacgaacg gtcctcttcg cagatcaggg
     2761 cgttggggcg gagtctcacc aaggactacg tctgctggcg atttccgtta caccccgggc
     2821 ggtggccggc gcacacgcgc gcccgcgttg ggcagtgcag aaagtgcaga aacctaggcg
     2881 ctgatggtcc aggtccacgg ttcgtcgtcg gcggcggcgc gggcggcggc gtcggccagg
     2941 gcgcgggcga gaccggctac ggcgggcttg atgcgccggt tgcgggcgac cttgagcagc
     3001 tagtatgcag gtcgacggat cttttccgct gcataaccct gcttcggggt cattatagcg
     3061 attttttcgg tatatccatc ctttttcgca cgatatacag gattttgcca aagggttcgt
     3121 gtagactttc cttggtgtat ccaacggcgt cagccgggca ggataggtga agtaggccca
     3181 cccgcgagcg ggtgttcctt cttcactgtc ccttattcgc acctggcggt gctcaacggg
     3241 aatcctgctc tgcgaggctg gccggctacc gccggcgtaa cagatgaggg caagcggatg
     3301 gctgatgaaa ccaagccaac caggaagggc agcccaccta tcaaggtgta ctgccttcca
     3361 gacgaacgaa gagcgattga ggaaaaggcg gcggcggccg gcatgagcct gtcggcctac
     3421 ctgctggccg tcggccaggg ctacaaaatc acgggcgtcg tggactatga gcacgtccgc
     3481 gagctggccc gcatcaatgg cgacctgggc cgcctgggcg gcctgctgaa actctggctc
     3541 accgacgacc cgcgcacggc gcggttcggt gatgccacga tcctcgccct gctggcgaag
     3601 atcgaagaga agcaggacga gcttggcaag gtcatgatgg gcgtggtccg cccgagggca
     3661 gagccatgac ttttttagcc gctaaaacgg ccggggggtg cgcgtgattg ccaagcacgt
     3721 ccccatgcgc tccatcaaga agagcgactt cgcggagctg gtgaagtaca tcaccgacga
     3781 gcaaggcaag accgatcccc ggggacctgc atagatctac gtctgtcgag aagtttctga
     3841 tygmmaagyw cgwcmscrts cgctccatca agaagagcga cttcgcggag ctggtgaagt
     3901 acatcaccga cgagcaaggc aagaccgatc cccggggacc tgcatagatc tacgtctgtc
     3961 gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct ctcggagggc
     4021 gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat
     4081 agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc atcggccgcg
     4141 ctcccgattc cggaagtgct tgacattggg gaatttatgc ggtgtgaaat accgcacaga
     4201 tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg
     4261 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     4321 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     4381 gccagtgcca agcttcgacc gccgctcgag ccgggcgccc aaagctgagc gcgtgaggga
     4441 ccacgaggca gggaggggaa cacatggcca cgcaggactc cgaagtcgca ctggtgacgg
     4501 gtgcgaccag cggaatcggg ctggagatcg cccgcaggct cggtaaggag gggttgcgcg
     4561 tgttcgtctg cgcgcgcggc gaagaaggac tgcggacgac gctgaaggag ttgcgggagg
     4621 cgggcgtgga ggcggacggg cggacctgcg acgtccgctc ggtcccggag atcgaggctc
     4681 tcgtggcggc ggtggtcgag cgttacggtc cggtcgacgt gctggtgaac aacgcgggcc
     4741 ggcccggcgg cggcgccacg gccgaactcg ccgacgaact ctggctcgat gtcgtggaga
     4801 ccaacctcac cggcgtgttc cgggtgacca aacaggtcct caaggcgggc ggcatgctcg
     4861 aacggggcac gggccgaatc gtcaacatcg cctcgaccgg cggaaagcag ggtgtggtgc
     4921 acgccgcgcc ctactccgcc tcgaagcacg gcgtggtcgg cttcaccaag gcgctcggtc
     4981 tcgaactggc gaggaccggc atcacggtga acgccgtctg ccccggattc gtcgagacgc
     5041 cgatggccgc gtccgtgcgc gagcactact cggacatctg ggaggtgtcg accgaggagg
     5101 ccttcgaccg gatcaccgcg cgcgtgccga tcggccggta cgtgcagccg tccgaggtgg
     5161 cggagatggt ggcgtacctg atcgggcccg gtgcggccgc gttcacggac cggtgctgcg
     5221 cccccgtcgr grtycwycgk cysstrsgkg crgccgtccg aggtggcgga gatggtggcg
     5281 tacctgatcg ggcccggtgc ggccgcgttc acggaccggt gctgcgcccc cgtcgagatt
     5341 ctccgtctcc tgggggcgac aataacaggc ctactcaaca gatttcaatg tgcttgccgt
     5401 cacaagcgat ctcctattga tcaaaaatta atcagtgcag cgagctgcac tgattaattt
     5461 ttgctggatt ttaccgagag gcagtggtcg aggcttccga atcaatagaa tctggcattc
     5521 ctgatacggc tgggtcctcg gaagggtcct cgcgccgggc gagcgggtgc ggccaccaca
     5581 tccaccggcc gacatcgaga gtgagcgcgg tgaccaggac cgatcgcacg atcatggtgt
     5641 cgagcaggac gcccaccgcg accgtgaatc cgagctcggc gatgaagacc agcggcagcg
     5701 tggccagcgc ggcgaaggtc ccggccagca ccagcccggc cgcggtgatc acgccaccgg
     5761 tcgaggtgag gcccttgagc gcgccctgcc gggtgccgag ccggtcgctc tcctccttga
     5821 tccgggtgac gaggaagatg ttgtagtcga cgcccagggc gaccaggaag acgaagaccc
     5881 acagggggaa ggccgaatcg gctcccgcga agtcgaagac gtggttgaag aagagggtgg
     5941 cgaggccgag cgcggtgaag aaggacagca ccacgctcgc gatcagcagc agcggcgcca
     6001 ccagtgctcg cagcagcagc gcgaggatgc agaacaccac ggcgaggatc accggaatga
     6061 tcagaccgcg gtcgcggctg gacgcctccc gcatgtcgtg cacgacggcg ctgctgccgc
     6121 ccacccgtgc ctgcgcaccg tccagccggg cgagtgtctc gcgggccgcg gtgaccgaac
     6181 gcatggccgc cggcgagtcc gccccggcgc cgagcgtcgc ctcgacatag gcgaggccgt
     6241 ccttcacctg tggtggtgcg acctcgatga cgccgggcac ggtctccaga gctgcgtgca
     6301 cctggtccgc cgaggcgccc ttggcgatga cgaccatggg ttcaccggag ccggccggga
     6361 agtgccgggt caggatctcc tcgcccacca ccgactccgg cttggtggaa ttcagatctc
     6421 aaaaaaagca ccgactcggt gccacttttt caagttgata acggactagc cttattttaa
     6481 cttgctattt ctagctctaa aacgaactcc ggtccctggt aatactagta ttatacctag
     6541 gactgagcta gctgtcaagg atccgcggcc gcgcgcgatc tagagcaacc gagcgttctg
     6601 aacaaatcca gatggagttc tgaggtcatt actggaccgg atcggggatc tgggctgagg
     6661 gagccgacgg cacgcggcgg ctcacggcgt ggcacgcgga acgtccgggc ttgcacctca
     6721 cgtcacgtga ggaggcagcg tggacggcgt cagagaaggg agcggacata tggacaagaa
     6781 gtactccatc ggcctggaca tcggcaccaa ctccgtcggc tgggccgtca tcacggacga
     6841 gtacaaggtc ccgtcgaaga agttcaaggt gctgggcaac acggaccgcc acagcatcaa
     6901 gaagaacctc atcggcgccc tcctgttcga cagcggcgag acggccgagg cgacccgcct
     6961 gaagcgcacg gcgcggcggc ggtacacgcg ccgcaagaac cgcatctgct acctccagga
     7021 gatcttctcc aacgagatgg cgaaggtcga cgactcgttc ttccaccggc tggaggagtc
     7081 cttcctggtc gaggaggaca agaagcacga gcggcacccc atcttcggca acatcgtgga
     7141 cgaggtggcc taccacgaga agtacccgac catctaccac ctccggaaga agctggtgga
     7201 ctcgaccgac aaggccgacc tgcgcctgat ctacctggcc ctcgcccaca tgatcaagtt
     7261 ccgcggccac ttcctcatcg agggcgacct gaacccggac aactcggacg tcgacaagct
     7321 gttcatccag ctcgtgcaga cctacaacca gctcttcgag gagaacccga tcaacgcctc
     7381 cggcgtggac gcgaaggcca tcctgtcggc ccgcctctcg aagtcccggc ggctcgagaa
     7441 cctgatcgcc cagctgccgg gcgagaagaa gaacggcctc ttcggcaacc tgatcgcgct
     7501 ctccctgggc ctgacgccga acttcaagtc caacttcgac ctcgccgagg acgccaagct
     7561 ccagctgtcc aaggacacgt acgacgacga cctggacaac ctcctggcgc agatcggcga
     7621 ccagtacgcg gacctgttcc tcgcggcgaa gaacctctcc gacgcgatcc tgctcagcga
     7681 catcctccgc gtcaacaccg agatcaccaa ggcgcccctg tccgcgagca tgatcaagcg
     7741 ctacgacgag caccaccagg acctcaccct gctgaaggcc ctggtccgcc agcagctccc
     7801 ggagaagtac aaggagatct tcttcgacca gtccaagaac ggctacgcgg gctacatcga
     7861 cggcggcgcg tcccaggagg agttctacaa gttcatcaag cccatcctgg agaagatgga
     7921 cggcaccgag gagctgctgg tcaagctgaa ccgggaggac ctcctgcgca agcagcgcac
     7981 gttcgacaac ggctccatcc cccaccagat ccacctgggc gagctgcacg ccatcctccg
     8041 gcgccaggag gacttctacc cgttcctcaa ggacaaccgc gagaagatcg agaagatcct
     8101 cacgttccgg atcccgtact acgtcggccc cctcgcccgc ggcaactccc gcttcgcgtg
     8161 gatgacgcgc aagtcggagg agacgatcac cccgtggaac ttcgaggagg tggtggacaa
     8221 gggcgcctcc gcgcagtcgt tcatcgagcg gatgaccaac ttcgacaaga acctgccgaa
     8281 cgagaaggtg ctgcccaagc acagcctcct gtacgagtac ttcacggtct acaacgagct
     8341 caccaaggtg aagtacgtga cggagggcat gcggaagccg gcgttcctgt ccggcgagca
     8401 gaagaaggcc atcgtggacc tgctgttcaa gacgaaccgg aaggtcaccg tcaagcagct
     8461 gaaggaggac tacttcaaga agatcgagtg cttcgactcg gtcgagatca gcggcgtgga
     8521 ggaccgcttc aacgcgtcgc tgggcacgta ccacgacctc ctgaagatca tcaaggacaa
     8581 ggacttcctc gacaacgagg agaacgagga catcctggag gacatcgtgc tgaccctgac
     8641 gctcttcgag gaccgggaga tgatcgagga gcgcctcaag acctacgcgc acctgttcga
     8701 cgacaaggtc atgaagcagc tcaagcggcg gcggtacacc ggctggggcc ggctctcccg
     8761 caagctcatc aacggcatcc gggacaagca gtcgggcaag accatcctcg acttcctgaa
     8821 gtccgacggc ttcgccaacc gcaacttcat gcagctgatc cacgacgaca gcctgacgtt
     8881 caaggaggac atccagaagg cgcaggtgtc gggccagggc gactccctgc acgagcacat
     8941 cgcgaacctg gccggcagcc cggccatcaa gaagggcatc ctgcagacgg tcaaggtggt
     9001 cgacgagctg gtgaaggtga tgggccgcca caagcccgag aacatcgtca tcgagatggc
     9061 ccgggagaac cagaccacgc agaagggcca gaagaacagc cgcgagcgca tgaagcggat
     9121 cgaggagggc atcaaggagc tcggctcgca gatcctcaag gagcaccccg tcgagaacac
     9181 ccagctccag aacgagaagc tctacctcta ctacctgcag aacggccggg acatgtacgt
     9241 ggaccaggag ctcgacatca accggctgtc ggactacgac gtcgaccaca tcgtcccgca
     9301 gtcgttcctg aaggacgaca gcatcgacaa caaggtcctc acccgctcgg acaagaaccg
     9361 gggcaagtcc gacaacgtcc cgtcggagga ggtcgtgaag aagatgaaga actactggcg
     9421 gcagctcctc aacgccaagc tcatcacgca gcgcaagttc gacaacctga cgaaggcgga
     9481 gcggggcggc ctcagcgagc tcgacaaggc gggcttcatc aagcggcagc tggtcgagac
     9541 gcggcagatc acgaagcacg tggcccagat cctggactcg cgcatgaaca ccaagtacga
     9601 cgagaacgac aagctgatcc gcgaggtcaa ggtcatcacc ctcaagtcca agctggtgag
     9661 cgacttccgc aaggacttcc agttctacaa ggtgcgggag atcaacaact accaccacgc
     9721 ccacgacgcg tacctgaacg cggtggtggg cacggcgctg atcaagaagt acccgaagct
     9781 cgagagcgag ttcgtctacg gcgactacaa ggtgtacgac gtgcggaaga tgatcgcgaa
     9841 gagcgagcag gagatcggca aggccacggc gaagtacttc ttctacagca acatcatgaa
     9901 cttcttcaag accgagatca ccctggcgaa cggcgagatc cgcaagcgcc cgctcatcga
     9961 gaccaacggc gagacgggcg agatcgtgtg ggacaagggc cgcgacttcg cgaccgtccg
    10021 gaaggtgctg tccatgcccc aggtcaacat cgtcaagaag acggaggtcc agaccggcgg
    10081 cttcagcaag gagtccatcc tgcccaagcg gaacagcgac aagctgatcg cccggaagaa
    10141 ggactgggac cccaagaagt acggcggctt cgactccccc accgtcgcct actcggtcct
    10201 cgtggtcgcc aaggtggaga agggcaagag caagaagctg aagtccgtga aggagctgct
    10261 gggcatcacc atcatggagc ggtcgtcgtt cgagaagaac cccatcgact tcctggaggc
    10321 caagggctac aaggaggtga agaaggacct catcatcaag ctccccaagt actccctgtt
    10381 cgagctggag aacggccgga agcgcatgct cgccagcgcg ggcgagctgc agaagggcaa
    10441 cgagctggcg ctgccgagca agtacgtcaa cttcctgtac ctcgcctcgc actacgagaa
    10501 gctgaagggc tcgccggagg acaacgagca gaagcagctg ttcgtggagc agcacaagca
    10561 ctacctcgac gagatcatcg agcagatctc cgagttctcc aagcgcgtca tcctggccga
    10621 cgcgaacctg gacaaggtcc tgtccgcgta caacaagcac cgcgacaagc cgatccggga
    10681 gcaggccgag aacatcatcc acctgttcac gctgaccaac ctcggcgccc ccgcggcctt
    10741 caagtacttc gacaccacga tcgaccggaa gcggtacacg agcacgaagg aggtgctcga
    10801 cgccaccctg atccaccaga gcatcacggg cctgtacgag acccgcatcg acctctccca
    10861 gctgggcggc gactgagaat tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta
    10921 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
    10981 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
    11041 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
    11101 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
    11161 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
    11221 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
    11281 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
    11341 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
    11401 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
    11461 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
    11521 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
    11581 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
    11641 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
    11701 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
    11761 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
    11821 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
    11881 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
    11941 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt tggttcatgt
    12001 gcagctccat cagcaaaagg ggatgataag tttatcacca ccgactattt gcaacagtgc
    12061 cgttgatcgt gctatgatcg actgatgtca tcagcggtgg agtgcaatgt cgtgcaatac
    12121 gaatggcgaa aagccgagct catcggtcag cttctcaacc ttggggttac ccccggcggt
    12181 gtgctgctgg tccacagctc cttccgtagc gtccggcccc tcgaagatgg gccacttgga
    12241 ctgatcgagg ccctgcgtgc tgcgctgggt ccgggaggga cgctcgtcat gccctcgtgg
    12301 tcaggtctgg acgacgagcc gttcgatcct gccacgtcgc ccgttacacc ggaccttgga
    12361 gttgtctctg acacattctg gcgcctgcca aatgtaaagc gcagcgccca tccatttgcc
    12421 tttgcggcag cggggccaca ggcagagcag atcatctctg atccattgcc cctgccacct
    12481 cactcgcctg caagcccggt cgcccgtgtc catgaactcg atgggcaggt acttctcctc
    12541 ggcgtgggac acgatgccaa cacgacgctg catcttgccg agttgatggc aaaggttccc
    12601 tatggggtgc cgagacactg caccattctt caggatggca agttggtacg cgtcgattat
    12661 ctcgagaatg accactgctg tgagcgcttt gccttggcgg acaggtggct caaggagaag
    12721 agccttcaga aggaaggtcc agtcggtcat gcctttgctc ggttgatccg ctcccgcgac
    12781 attgtggcga cagccctggg tcaactgggc cgagatccgt tgatcttcct gcatccgcca
    12841 gaggcgggat gcgaagaatg cgatgccgct cgccagtcga ttggctgagc tcatgagcgg
    12901 agaacgagat gacgttggag gggcaaggtc gcgctgattg ctggggcaac acgtggagcg
    12961 gatcggggat tgtctttctt cagctcgctg atgatatgct gacgctcaat gccgtttggc
    13021 ctccgactaa cgaaaatccc gcatttggac ggctgatccg attggcacgg cggacggcga
    13081 atggcggagc agacgctcgt ccgggggcaa tgagatatga aaaagcctga actcaccgcg
    13141 acgtatcgat gtcgaggttc ctcaggggag ccaccccaga gaagccctcg gagctgagcg
    13201 gagctatttc caaagccatg ccagctagag acagtgcaca ccgccaagca tctgcaaaac
    13261 cctcgcctgg agggaaagtg caatgtacgt acgctggttt ccctccagaa agggattctg
    13321 cggcttctca actttggaag aagaggcgga acgaactgct gtttcagcct actca
//
