LOCUS       sequence-262065-       14529 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gene knockout/in for actinomycetes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14529)
  AUTHORS   Tong Y, Charusanti P, Zhang L, Weber T, Lee SY
  TITLE     CRISPR-Cas9 Based Engineering of Actinomycetal Genomes.
  JOURNAL   ACS Synth Biol. 2015 Sep 18;4(9):1020-9. doi: 
            10.1021/acssynbio.5b00038. Epub 2015 Apr 7.
  PUBMED    25806970
REFERENCE   2  (bases 1 to 14529)
  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: 
            "10.1021/acssynbio.5b00038"; journalName: "ACS Synth Biol"; date: 
            "2015-09-18- 18"; volume: "4"; issue: "9"; pages: "1020-9"
COMMENT     Sequence Label: pCRISPR-Cas9-ScaligD
FEATURES             Location/Qualifiers
     source          1..14529
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            466..575
                     /note="incP origin of transfer"
     CDS             891..1691
                     /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      1993..2580
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2481..2500
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2734..2751
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2868..2889
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2904..2909,2910..2927,2928..2934)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2942..2958
                     /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)."
     primer_bind     2947..2969
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2966..2982
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2966..2982
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     misc_RNA        3120..3195
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      complement(3365..3399)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             3567..7673
                     /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             3567..7670
                     /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"
     mobile_element  7877..8644
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     terminator      complement(8644..8682)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             12891..14333
                     /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"
ORIGIN
        1 ggccgtcgac gacgacgcgt ccgcctgcct gcttttggcc ggcgggccgg gccgcggttg
       61 tccgccgggt cggaccggtc ccttggtcga ccgatgcctc ggccacgacg atgctgggcc
      121 cgaagcggta cgcgacccgt gccgggccgc agcggatcgc ggcagcagga gctccccgcc
      181 cgccacggct ggctggatgg tcgcctcacg cgcccgcagg ggtgacggcg tgggtgcggc
      241 ggtcagcccc gtgatcgggc gtcagacggc cgctgaggga ccgcgaccac ccctcccttc
      301 ccgcctgccg tctttcaccg cccggccggt ccgcagacgc cgtacggcca ggccacggag
      361 gattacgggc agatggcgcc cacccggcac ccgggctggg gcaccctgga cagcacgaag
      421 gtccgcccca ctcaccgggg cgggccttcg cgttgagcta gagttcccgc cagcctcgca
      481 gagcaggatt cccgttgagc accgccaggt gcgaataagg gacagtgaag aaggaacacc
      541 cgctcgcggg tgggcctact tcacctatcc tgcccggctg acgccgttgg atacaccaag
      601 gaaagtctac acgaaccctt tggcaaaatc ctgtatatcg tgcgaaaaag gatggatata
      661 ccgaaaaaat cgctataatg accccgaagc agggttatgc agcggaaaat gcagctcacg
      721 gtaactgatg ccgtatttgc agtaccagcg tacggcccac agaatgatgt cacgctgaaa
      781 atgccggcct ttgaatgggt tcatgtgcag ctccataagc aaaaggggat gataagttta
      841 tcaccaccga ctatttgcaa cagtgccgtt gatcgtgcta tgatcgactg atgtcatcag
      901 cggtggagtg caatgtcgtg caatacgaat ggcgaaaagc cgagctcatc ggtcagcttc
      961 tcaaccttgg ggttaccccc ggcggtgtgc tgctggtcca cagctccttc cgtagcgtcc
     1021 ggcccctcga agatgggcca cttggactga tcgaggccct gcgtgctgcg ctgggtccgg
     1081 gagggacgct cgtcatgccc tcgtggtcag gtctggacga cgagccgttc gatcctgcca
     1141 cgtcgcccgt tacaccggac cttggagttg tctctgactc attctggcgc ctgccaaatg
     1201 taaagcgcag cgcccatcca tttgcctttg cggcagcggg gccacaggca gagcagatca
     1261 tctctgatcc attgcccctg ccacctcact cgcctgcaag cccggtcgcc cgtgtccatg
     1321 aactcgatgg gcaggtactt ctcctcggcg tgggacacga tgccaacacg acgctgcatc
     1381 ttgccgagtt gttggcaaag gttccctatg gggtgccgag acactgcacc attcttcagg
     1441 atggcaagtt ggtacgcgtc gattatctcg agaatgacca ctgctgtgag cgctttgcct
     1501 tggcggacag gtggctcaag gagaagagcc ttcagaagga aggtccagtc ggtcatgcct
     1561 ttgctcggtt gatccgctcc cgcgacattg tggcgacagc cctgggtcaa ctgggccgag
     1621 atccgttgat cttcctgcat ccgccagagg cgggatgcga agaatgcgat gccgctcgcc
     1681 agtcgattgg ctgagctcat aagttcctat tccgaagttc ctattcgaaa tgaccgacca
     1741 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     1801 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     1861 gctggagttc ttcgcccacc ccaaaaggat ctaggtgaag atcctttttg ataatctcat
     1921 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     1981 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2041 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     2101 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     2161 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     2221 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     2281 gttaccggat aaggcgcagc ggtcgggctg aacggggggt cgtgcacaca gcccagcttg
     2341 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     2401 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     2461 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     2521 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     2581 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     2641 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     2701 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     2761 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     2821 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     2881 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     2941 attgtgagcg gataacaatt tcacacagga aacagctatg acatgattac gaattgtacg
     3001 cggtcgatct tgacggctgg cgagaggtgc ggggaggatc tgaccgacgc ggtccacacg
     3061 tggcaccgcg atgctgttgt gggcacaatc gtgccggttg gtaggatcga cggccatggg
     3121 ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg
     3181 gcaccgagtc ggtgcttttt ttacgtaggc gtcagctacg atgttccggg gactgctgat
     3241 ccggtcagca ggtggaagag ggactggatt ccaaagttct caatgctgct tgctgttctt
     3301 gaatgggggg tcgttgacga cgacatggct cgattggcgc gacaagttgc tgcgattctc
     3361 accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat ggagttctga
     3421 ggtcattact ggaccggatc ggggatctgg gctgagggag ccgacggcac gcggcggctc
     3481 acggcgtggc acgcggaacg tccgggcttg cacctcacgt cacgtgagga ggcagcgtgg
     3541 acggcgtcag agaagggagc ggacatatgg acaagaagta ctccatcggc ctcgacatcg
     3601 gcaccaactc cgtgggctgg gcggtcatca ccgacgagta caaggtcccc tccaagaagt
     3661 tcaaggtcct gggcaacacc gaccggcact cgatcaagaa gaacctgatc ggcgccctgc
     3721 tcttcgacag cggcgagacc gccgaggcga cccgcctgaa gcggaccgcg cgtcgccgct
     3781 acacccggcg caagaaccgc atctgctacc tgcaggaaat cttctccaac gagatggcca
     3841 aggtggacga ctcgttcttc caccgcctgg aggagagctt cctggtggag gaggacaaga
     3901 agcacgagcg ccacccgatc ttcggcaaca tcgtggacga ggtggcctac cacgagaagt
     3961 accccaccat ctaccacctc cgcaagaagc tggtggactc gaccgacaag gcggacctgc
     4021 ggctcatcta cctggccctc gcgcacatga tcaagttccg cggccacttc ctcatcgagg
     4081 gcgacctgaa cccggacaac tccgacgtgg acaagctctt catccagctg gtgcagacct
     4141 acaaccagct gttcgaggag aaccccatca acgccagcgg cgtggacgcc aaggcgatcc
     4201 tctccgcgcg cctgagcaag tcccggcgcc tggagaacct catcgcccag ctgccgggcg
     4261 agaagaagaa cggcctcttc ggcaacctga tcgcgctgtc gctcggcctg acccccaact
     4321 tcaagagcaa cttcgacctg gccgaggacg cgaagctcca gctgtccaag gacacctacg
     4381 acgacgacct ggacaacctg ctcgcccaga tcggcgacca gtacgcggac ctcttcctgg
     4441 ccgcgaagaa cctctcggac gccatcctgc tcagcgacat cctgcgggtc aacaccgaga
     4501 tcaccaaggc cccgctgtcg gcgagcatga tcaagcggta cgacgagcac caccaggacc
     4561 tgaccctgct caaggccctc gtgcgccagc agctgcccga gaagtacaag gaaatcttct
     4621 tcgaccagtc caagaacggc tacgccggct acatcgacgg cggcgcgtcg caggaggagt
     4681 tctacaagtt catcaagccg atcctggaga agatggacgg caccgaggag ctgctcgtca
     4741 agctgaaccg cgaggacctg ctccgcaagc agcggacctt cgacaacggc tccatcccgc
     4801 accagatcca cctgggcgag ctccacgcca tcctccggcg ccaggaggac ttctacccct
     4861 tcctgaagga caaccgcgag aagatcgaga agatcctgac cttccgcatc ccgtactacg
     4921 tcggccccct ggcccgcggc aactcccggt tcgcgtggat gacccggaag tcggaggaga
     4981 ccatcacccc gtggaacttc gaggaggtcg tggacaaggg cgcgtccgcg cagtcgttca
     5041 tcgagcgcat gaccaacttc gacaagaacc tcccgaacga gaaggtcctg cccaagcact
     5101 ccctgctcta cgagtacttc accgtgtaca acgagctgac caaggtcaag tacgtgaccg
     5161 agggcatgcg gaagccggcc ttcctgtcgg gcgagcagaa gaaggcgatc gtggacctgc
     5221 tcttcaagac caaccgcaag gtcaccgtga agcagctgaa ggaggactac ttcaagaaga
     5281 tcgagtgctt cgactccgtc gagatcagcg gcgtggagga ccgcttcaac gcctccctgg
     5341 gcacctacca cgacctgctc aagatcatca aggacaagga cttcctcgac aacgaggaga
     5401 acgaggacat cctggaggac atcgtcctca ccctgaccct cttcgaggac cgcgagatga
     5461 tcgaggagcg gctcaagacc tacgcccacc tgttcgacga caaggtgatg aagcagctga
     5521 agcgtcgccg ctacaccggc tggggccgcc tctcccggaa gctgatcaac ggcatccggg
     5581 acaagcagag cggcaagacc atcctggact tcctcaagtc cgacggcttc gccaaccgca
     5641 acttcatgca gctcatccac gacgacagcc tgaccttcaa ggaggacatc cagaaggccc
     5701 aggtctcggg ccagggcgac agcctccacg agcacatcgc caacctggcg ggctccccgg
     5761 cgatcaagaa gggcatcctc cagaccgtca aggtcgtgga cgagctggtc aaggtgatgg
     5821 gccgccacaa gcccgagaac atcgtgatcg agatggcccg ggagaaccag accacccaga
     5881 agggccagaa gaactcgcgc gagcggatga agcggatcga ggagggcatc aaggagctcg
     5941 gcagccagat cctgaaggag cacccggtcg agaacaccca gctgcagaac gagaagctgt
     6001 acctctacta cctgcagaac ggccgcgaca tgtacgtgga ccaggagctc gacatcaacc
     6061 ggctgtccga ctacgacgtg gaccacatcg tgccgcagtc cttcctgaag gacgactcga
     6121 tcgacaacaa ggtcctgacc cgctcggaca agaaccgggg caagtccgac aacgtgccct
     6181 cggaggaggt cgtgaagaag atgaagaact actggcgcca gctgctcaac gccaagctca
     6241 tcacccagcg caagttcgac aacctgacca aggccgagcg gggcggcctg agcgagctcg
     6301 acaaggcggg cttcatcaag cgccagctgg tcgagacccg gcagatcacc aagcacgtgg
     6361 cccagatcct ggactcccgg atgaacacca agtacgacga gaacgacaag ctgatccgcg
     6421 aggtcaaggt gatcaccctc aagagcaagc tggtctccga cttccgcaag gacttccagt
     6481 tctacaaggt ccgggagatc aacaactacc accacgccca cgacgcgtac ctgaacgccg
     6541 tcgtgggcac cgcgctgatc aagaagtacc cgaagctgga gtccgagttc gtctacggcg
     6601 actacaaggt ctacgacgtg cgcaagatga tcgccaagag cgagcaggag atcggcaagg
     6661 ccaccgcgaa gtacttcttc tactccaaca tcatgaactt cttcaagacc gagatcaccc
     6721 tggccaacgg cgagatccgc aagcggcccc tgatcgagac caacggcgag accggcgaga
     6781 tcgtctggga caagggccgc gacttcgcca ccgtccggaa ggtgctgtcg atgccgcagg
     6841 tcaacatcgt gaagaagacc gaggtgcaga ccggcggctt cagcaaggag tccatcctcc
     6901 ccaagcgcaa cagcgacaag ctgatcgccc ggaagaagga ctgggacccg aagaagtacg
     6961 gcggcttcga cagccccacc gtcgcctact ccgtgctggt cgtggcgaag gtcgagaagg
     7021 gcaagagcaa gaagctgaag tccgtgaagg agctgctcgg catcaccatc atggagcgct
     7081 cctcgttcga gaagaacccg atcgacttcc tggaggccaa gggctacaag gaggtcaaga
     7141 aggacctcat catcaagctg cccaagtaca gcctgttcga gctggagaac ggccgcaagc
     7201 ggatgctcgc ctccgcgggc gagctgcaga agggcaacga gctggccctc ccgtcgaagt
     7261 acgtcaactt cctgtacctc gcgtcccact acgagaagct gaagggctcg cccgaggaca
     7321 acgagcagaa gcagctcttc gtggagcagc acaagcacta cctggacgag atcatcgagc
     7381 agatcagcga gttcagcaag cgcgtcatcc tggccgacgc gaacctcgac aaggtgctgt
     7441 ccgcctacaa caagcaccgc gacaagccga tccgggagca ggcggagaac atcatccacc
     7501 tgttcaccct caccaacctg ggcgcccccg ccgcgttcaa gtacttcgac accaccatcg
     7561 accgcaagcg gtacacctcc accaaggagg tcctcgacgc gaccctgatc caccagagca
     7621 tcaccggcct gtacgagacc cgcatcgacc tgtcccagct cggcggcgac tgaaagcttt
     7681 ctagaggatc cccgggtacc gagctcgaat tccccagatc taaagttttg tcgtctttcc
     7741 agacgttagt aaatgaattt tctgtatgag gttttgctaa acaactttca acagtttcag
     7801 cggagtgaga atagaaagga acaactaaag gaattgcgaa taataatttt ttcacgttga
     7861 aaatctccaa aaaaaaggta atgactccaa cttattgata gtgttttatg ttcagataat
     7921 gcccgatgac tttgtcatgc agctccaccg attttgagaa cgacagcgac ttccgtccca
     7981 gccgtgccag gtgctgcctc agattcaggt tatgccgctc aattcgctgc gtatatcgct
     8041 tgctgattac gtgcagcttt cccttcaggc gggattcata cagcggccag ccatccgtca
     8101 tccatatcac cacgtcaaag ggtgacagca ggctcataag acgccccagc gtcgccatag
     8161 tgcgttcacc gaatacgtgc gcaacaaccg tcttccggag actgtcatac gcgtaaaaca
     8221 gccagcgctg gcgcgattta gccccgacat agccccactg ttcgtccatt tccgcgcaga
     8281 cgatgacgtc actgcccggc tgtatgcgcg aggttaccga ctgcggcctg agttttttaa
     8341 gtgacgtaaa atcgtgttga ggccaacgcc cataatgcgg gctgttgccc ggcatccaac
     8401 gccattcatg gccatatcaa tgattttctg gtgcgtaccg ggttgagaag cggtgtaagt
     8461 gaactgcagt tgccatgttt tacggcagtg agagcagaga tagcgctgat gtccggcggt
     8521 gcttttgccg ttacgcacca ccccgtcagt agctgaacag gagggacagc tgatagaaac
     8581 agaagccact ggagcacctc aaaaacacca tcatacacta aatcagtaag ttggcagcat
     8641 cacccaaaaa aaaaggctcc aaaaggagcc tttaattgta tcggtttatc agcttgcttt
     8701 cgaggtgaat ttcttaaaca gcttgatacc gatagttgcg ccgacaatga caacaaccat
     8761 cgcccacgca taaccgatat attcggtcgc tgaggcttgc agggagtcaa aggccgcttt
     8821 tgcgggatct cgtcgaaggc actagaaggg cggtcgatct tgacggctgg cgagaggtgc
     8881 ggggaggatc tgaccgacgc ggtccacacg tggcaccgcg atgctgttgt gggcacaatc
     8941 gtgccggttg gtaggatcga cggcccggcg tgcaccgggc gttcggcggt gaaccggggc
     9001 agccggatat cgaggtccgg ctgagcaacc tggacaaggt gctctatcca gcgaccggca
     9061 ccaccaaggg cgaggtcatc gagtactacg ccgaaatcgc cccggcgatg ctgccgcata
     9121 tcgcgggccg gccgatcacc cggaaacggt ggccgaacgg tgtcgccgaa tcgtcgttct
     9181 tcgagaagaa cctcggcgcg ggtacaccgt cgtggctacc gcgccgtgcc caggaacatt
     9241 ccgaccgcac cgcgcactat ccggtgatct cgtcgcaggc cggcctggtc tggctgggtc
     9301 agcaggccgc cctggagatc cacgtaccgc aatggcgctt cgacggcgat gcgcgcggac
     9361 ccgcgacgcg gctggtgttc gatctcgatc ccggccccgg cgcgggactg cccgaatgcg
     9421 cgcgggtggc gctcggggtg cgggatatgg tcgccgaaat cgggatgcgc gcgttcccgc
     9481 tgaccagcgg tagcaaaggt atccacctgt acgtcccgct ggaccgggtg ctgagccccg
     9541 gcggggcgtc cacggtggcc aaacaggtcg ccgcgaatct ggagaaactc cttcccgacc
     9601 tggtcaccgc cacgatggcg aagagtgtgc gggccgggaa ggtgttcctg gactggagtc
     9661 agaacaaccc gtccaagacg accatcgcac cgtattcgct gcgcggccgc gagcagccga
     9721 acgtcgccgc accacgccac tgggcggagc tcgaggacgc ccgtgaactg cggcagctgc
     9781 ggttcgacga agttctggag cgttatcggt ccgagggtga tctgctggcc ggcctggata
     9841 cacccctgaa cgacgcgttg acgaaatacc gatcgatgcg tgacccggcg cgtacaccgg
     9901 agccggtacc gccgcattcg ccccggcccg gccccggtga ccgctatgtc gtccacgaac
     9961 accacgcccg gcggttgcac tgggatgtgc ggttggaacg cgacggggtg ctggtgtcgt
    10021 gggcggtgcc caaggggccg ccggaaagca cccggcagaa tcggctcgcc gtgcacaccg
    10081 aggaccaccc gctggaatac ctggacttcc acggcacgat cccggccggc gagtacgggg
    10141 caggggagct gtcggtctgg gataccggca cctaccgcgc cgagaaatgg cgcgacgacg
    10201 aggtgatcgt ggttttccgg ggcgagcggc tcaacggccg gtacgccatg atccggaccg
    10261 agggcgatca atggctgatg catctcatga aggaccagcc cgcgaccggg gaactgccgc
    10321 gtggactcac ccccatgctg gccaccagtg gcgaagtggc cgggctgccg gactcggagt
    10381 gggcgttcga acgtaaatgg gacggatacc ggctgctcgt cgaaatcgat gccggcgaaa
    10441 tgcggctgcg cagccgggcc ggtaacgacg tcaccgcgcg ctatccccag ttgtcggtgc
    10501 tggccgagga gctggccgac catcaggtga tactcgacgg tgagctcatc gtccgcggcc
    10561 ccgacggcgc ggtgaatatc gcgctgttga aggcgaatcc gcggcgcgcc gaattcctgg
    10621 cgttcgatct gctgttcctc gacggcactt cactgctgcg caaacgctac cgcgatcggc
    10681 ggcacgtgct cgaagcgctg gccgcgacca ccaccgaact ccgggtgcca ccgcgctatg
    10741 agggcgacgg caccgaggcc ctgcaccgca gcgaagaaga tggcgccgag ggcgtgatcg
    10801 ccaaacggct ggattcggtg tatctgcccg ggacccgcgg gcattcgtgg gtgaagcacc
    10861 ggaactggcg tacccaggag gtggtgatcg ggggtatgcg gcgcagtaag gcgcgaccgt
    10921 tcgcctcgtt gctggtcggg ataccggccg aggacggcct ggtgtatgcg ggccgggtcg
    10981 ggaccgggtt cgacgaagcg gggatgaccg aactcgcggc ccggctgcgc cggtcggaac
    11041 gtaagacgcc gccgttcacc aacgagatgt cggccgatga actccgggac gcgatctggg
    11101 tgacaccgaa gatcaaaggc actgttcgct acatggattg gaccgacggc ggacgcttct
    11161 ggcatcctgc ctggctcggc gaggtgtgac agccgtcgct cagcaggtgg aagagggact
    11221 ggattccaaa gttctcaatg ctgcttgctg ttcttgaatg gggggtcgtt gacgacgaca
    11281 tggctcgatt ggcgcgacaa gttgctgcga ttctcaccaa taaaaaacgc ccggcggcac
    11341 ctatatgcgg ggatcgaccg cgcgggtccc ggacggggaa gagcggggag ctttgccaga
    11401 gagcgacgac ttccccttgc gttggtgatt gccggtcagg gcagccatcc gccatcgtcg
    11461 cgtagggtgt cacaccccag gaatcgcgtc actgaacaca gcagccggta ggacgaccat
    11521 gactgagttg gacaccatcg caaatccgtc cgatcccgcg gtgcagcgga tcatcgatgt
    11581 caccaagccg tcgcgatcca acataaagac aacgttgatc gaggacgtcg agcccctcat
    11641 gcacagcatc gcggccgggg tggagttcat cgaggtctac ggcagcgaca gcagtccttt
    11701 tccatctgag ttgctggatc tgtgcgggcg gcagaacata ccggtccgcc tcatcgactc
    11761 ctcgatcgtc aaccagttgt tcaaggggga gcggaaggcc aagacattcg gcatcgcccg
    11821 cgtccctcgc ccggccaggt tcggcgatat cgcgagccgg cgtggggacg tcgtcgttct
    11881 cgacggggtg aagatcgtcg ggaacatcgg cgcgatagta cgcacgtcgc tcgcgctcgg
    11941 agcgtcgggg atcatcctgg tcgacagtga catcaccagc atcgcggacc ggcgtctcca
    12001 aagggccagc cgaggttacg tcttctccct tcccgtcgtt ctctccggtc gcgaggaggc
    12061 catcgccttc attcgggaca gcggtatgca gctgatgacg ctcaaggcgg atggcgacat
    12121 ttccgtgaag gaactcgggg acaatccgga tcggctggcc ttgctgttcg gcagcgaaaa
    12181 gggtgggcct tccgacctgt tcgaggaggc gtcttccgcc tcggtttcca tccccatgat
    12241 gagccagacc gagtctctca acgtttccgt ttccctcgga atcgcgctgc acgagaggat
    12301 cgacaggaat ctcgcggcca accgataagc gcctctgttc ctcggacgct cggttgctcg
    12361 acctcgattc gtcagtgatg atcaacagga cccagaaatg gcacgagccc gggattggcc
    12421 tcccgggctc gccgcccccg aaccggcgca gcatgccgag tgaggacgtt tgctctgtga
    12481 cgagcgtacg agagaacagc gcgtcgacac caccgcgccc cgccggaccg aagtgccact
    12541 gcggcacccc gatcgtccac atccccggga agcgccccaa aatctactgc tcggacgcct
    12601 gccgaaagcg cgcaaagcgt ctcattgcgg accagtcacg cagcgtagca agcacgacta
    12661 cagcgggtaa tccggacgcg ggctttagta gtaagggaaa atctccgtcc actgcaggtc
    12721 agagcgctgt aactggaacg ggttctaccc caaacgaggg tgccgcgaca gccgggcggg
    12781 aggacccgcg cgacgagcgg ttcgcccggc gggaccggca ccagacggtg tcgctgaacg
    12841 aggcgttcaa gggctgcggg acccggctca ctgccgggac ggccgagctg atgtggaagc
    12901 cgggcgaggc gacgtggggc aacacgtgcc gctgcaacaa cgtccacacc tgcccgtggt
    12961 gcatgagccg catcctggcc gtacggggct cgaacgtgca gctcgccgcc gacggtctcg
    13021 cggacgccgg gtacgggctg caccttggca cgaacacgtt gcgccacttc gagcgcatgg
    13081 cgttcggcac cgtccgcaag ggaatgcggc acggcctggt cgcggtcttg cacgacggct
    13141 ggaagggcgc gtacggctcg tccggccgcc gctggcgcac gatgcgcgac gacttcggga
    13201 tcatcggcta cgagcgggcc ttcgaggaca ccttcggctg gggctccggc tggcacctgc
    13261 actggcacac gctctgggtc acccgcgagg tgctcggccc ggacgcccag gccgcgttcc
    13321 gcgacgcact cgccggagcc tgggccgccg gggtcgaatc cgcgggcggc tacaccgtca
    13381 gcgagacgtg cgaccggccc ggttgctcct gtgagggcaa gggccacggc accgacgtgc
    13441 gcccactcaa cggcgcggac gccgctgacg gcgatgcggg caagcaggcc cgctacctct
    13501 acaaggacgg cgacaagacc aagggcggcg tcgccaagat cggcctggag ctggccggac
    13561 agaacttcaa ggccggtcgc ggcgacgacc gcatgggacc gctcgacctg ggcgacgcgg
    13621 cggccgccga gctgcagcgg cttcgtcggc ccggcccgtt cgtcgagaag taccgcgagc
    13681 gcgagttcgg cgtcttccag gtccgcaagc actaccggtc gcagaacctg aaccggctga
    13741 tcaaggaact cggcatccag caggacgtcc gcaccgagga ggagatcacc gacgacaccg
    13801 agggcctggt cgccatcgcg gtcatcccgg cctacatctg gtaccgatac atcgcccgcg
    13861 tcgccggacg tcgcctcgac ctcatcaagg tcgccgagac gtacggccta ccgggcgtcc
    13921 ggcggctcgt cgagtcctgg ggcctggtgt ggggcaagga cgtcctggac ccgcccgccc
    13981 cggaagcgcc ggccgcgccg ggcgaccttg acgctgacca gatgaggttc gaggtcatgt
    14041 ccgaggagga agctgctttc cgcgaggcgc gccggaaggc caacgaggcc cgcacggagg
    14101 agctggccgc ttcgctcgac cgagtgcgtc agccgaagaa ggaggcgatc cgcccgacga
    14161 tcagccttcg gaagcgcctg aagccgaagc cggtcaccgt cgacgtcaag actcccccgc
    14221 ccggcgccgc gtcgcccgtg tgccgccgct gcaagggcaa gctcgccccg gtactccagc
    14281 cctggggcca gcaccccgga gactgcctcc gcgtcgacac agcggtcgcg tagggccgct
    14341 gagggaccgc catgagaccg ccaccccgga catccgggtg gcggtctcat ggcggtcccg
    14401 gcgcgaaagc cgagggtctg cctgccgtga ggtggccggc ggacctgggt tactgctgga
    14461 atcccgcgcc cagtccgagc gtccgccggc gcttttcacc tggctccggg tgctcgtcga
    14521 tgagcacca
//
