LOCUS       sequence-190162-       10994 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Streptomyces expression of codon-optimized Cas9 and custom gRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10994)
  AUTHORS   Cobb RE, Wang Y, Zhao H
  TITLE     High-Efficiency Multiplex Genome Editing of Streptomyces Species 
            Using an Engineered CRISPR/Cas System.
  JOURNAL   ACS Synth Biol. 2014 Dec 8.
  PUBMED    25458909
REFERENCE   2  (bases 1 to 10994)
  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: "ACS Synth 
            Biol. 2014 Dec 8."
COMMENT     Sequence Label: pCRISPomyces-2
FEATURES             Location/Qualifiers
     source          1..10994
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(153..1595)
                     /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            2762..2871
                     /note="incP origin of transfer"
     promoter        3093..3394
                     /label=rpsL promoter
                     /note="promoter for the Xylanimonas cellulosilytica 
                     ribosomal protein S12 gene"
     CDS             3395..7501
                     /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             3395..7498
                     /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"
     promoter        7701..7985
                     /label=gapdh promoter
                     /note="glyceraldehyde-3-phosphate dehydrogenase promoter 
                     from Eggerthella lenta"
     CDS             complement(7994..8290)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSG"
     primer_bind     8140..8162
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8154..8171
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8155..8171
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8181..8200
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8181..8199
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8247..8267)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(8247..8265)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8286..8302)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8286..8302)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8299..8321)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8310..8326
                     /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(8334..8340,8341..8358,8359..8364))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8379..8400
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_RNA        8443..8518
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      8540..8574
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     rep_origin      complement(8616..9204)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8696..8715)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             9408..10208
                     /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 cggacgctcg gactgggcgc gggattccag cagtaaccca ggtccgccgg ccacctcacg
       61 gcaggcagac cctcggcttt cgcgccggga ccgccatgag accgccaccc ggatgtccgg
      121 ggtggcggtc tcatggcggt ccctcagcgg ccctacgcga ccgctgtgtc gacgcggagg
      181 cagtctccgg ggtgctggcc ccagggctgg agtaccgggg cgagcttgcc cttgcagcgg
      241 cggcacacgg gcgacgcggc gccgggcggg ggagtcttga cgtcgacggt gaccggcttc
      301 ggcttcaggc gcttccgaag gctgatcgtc gggcggatcg cctccttctt cggctgacgc
      361 actcggtcga gcgaagcggc cagctcctcc gtgcgggcct cgttggcctt ccggcgcgcc
      421 tcgcggaaag cagcttcctc ctcggacatg acctcgaacc tcatctggtc agcgtcaagg
      481 tcgcccggcg cggccggcgc ttccggggcg ggcgggtcca ggacgtcctt gccccacacc
      541 aggccccagg actcgacgag ccgccggacg cccggtaggc cgtacgtctc ggcgaccttg
      601 atgaggtcga ggcgacgtcc ggcgacgcgg gcgatgtatc ggtaccagat gtaggccggg
      661 atgaccgcga tggcgaccag gccctcggtg tcgtcggtga tctcctcctc ggtgcggacg
      721 tcctgctgga tgccgagttc cttgatcagc cggttcaggt tctgcgaccg gtagtgcttg
      781 cggacctgga acacgccgaa ctcgcgctcg cggtacttct cgacgaacgg gccgggccga
      841 cgaagccgct gcagctcggc ggccgccgcg tcgcccaggt cgagcggtcc catgcggtcg
      901 tcgccgcgac cggccttgaa gttctgtccg gccagctcca ggccgatctt ggcgacgccg
      961 cccttggtct tgtcgccgtc cttgtagagg tagcgggcct gcttgcccgc atcgccgtca
     1021 gcggcgtccg cgccgttgag tgggcgcacg tcggtgccgt ggcccttgcc ctcacaggag
     1081 caaccgggcc ggtcgcacgt ctcgctgacg gtgtagccgc ccgcggattc gaccccggcg
     1141 gcccaggctc cggcgagtgc gtcgcggaac gcggcctggg cgtccgggcc gagcacctcg
     1201 cgggtgaccc agagcgtgtg ccagtgcagg tgccagccgg agccccagcc gaaggtgtcc
     1261 tcgaaggccc gctcgtagcc gatgatcccg aagtcgtcgc gcatcgtgcg ccagcggcgg
     1321 ccggacgagc cgtacgcgcc cttccagccg tcgtgcaaga ccgcgaccag gccgtgccgc
     1381 attcccttgc ggacggtgcc gaacgccatg cgctcgaagt ggcgcaacgt gttcgtgcca
     1441 aggtgcagcc cgtacccggc gtccgcgaga ccgtcggcgg cgagctgcac gttcgagccc
     1501 cgtacggcca ggatgcggct catgcaccac gggcaggtgt ggacgttgtt gcagcggcac
     1561 gtgttgcccc acgtcgcctc gcccggcttc cacatcagct cggccgtccc ggcagtgagc
     1621 cgggtcccgc agcccttgaa cgcctcgttc agcgacaccg tctggtgccg gtcccgccgg
     1681 gcgaaccgct cgtcgcgcgg gtcctcccgc ccggctgtcg cggcaccctc gtttggggta
     1741 gaacccgttc cagttacagc gctctgacct gcagtggacg gagattttcc cttactacta
     1801 aagcccgcgt ccggattacc cgctgtagtc gtgcttgcta cgctgcgtga ctggtccgca
     1861 atgagacgct ttgcgcgctt tcggcaggcg tccgagcagt agattttggg gcgcttcccg
     1921 gggatgtgga cgatcggggt gccgcagtgg cacttcggtc cggcggggcg cggtggtgtc
     1981 gacgcgctgt tctctcgtac gctcgtcaca gagcaaacgt cctcactcgg catgctgcgc
     2041 cggttcgggg gcggcgagcc cgggaggcca atcccgggct cgtgccattt ctgggtcctg
     2101 ttgatcctgg cattggtgtg gccgttcatt gcccctgctc gctcctgacg cgccgataga
     2161 cgtccgatac gcccggtgct ggtgggattt gataggtcgg aagaagcccc gcccgggcct
     2221 gggcggggct tcctgtgcgt caggacctcc tcgtcgtgag cctcttcggc ctatggacgg
     2281 agtgacctcg tgatccgtta cagccgcgcg cgctcgcgta gagcggtctc atcagttcca
     2341 cgaacggtcc tcttcgcaga tcagggcgtt ggggcggagt ctcaccaagg actacgtctg
     2401 ctggcgattt ccgttacacc ccgggcggtg gccggcgcac acgcgcgccc gcgttgggca
     2461 gtgcagaaag tgcagaaacc taggcgctga tggtccaggt ccacggttcg tcgtcggcgg
     2521 cggcgcgggc ggcggcgtcg gccagggcgc gggcgagacc ggctacggcg ggcttgatgc
     2581 gccggttgcg ggcgaccttg agcagctagt atgcaggtcg acggatcttt tccgctgcat
     2641 aaccctgctt cggggtcatt atagcgattt tttcggtata tccatccttt ttcgcacgat
     2701 atacaggatt ttgccaaagg gttcgtgtag actttccttg gtgtatccaa cggcgtcagc
     2761 cgggcaggat aggtgaagta ggcccacccg cgagcgggtg ttccttcttc actgtccctt
     2821 attcgcacct ggcggtgctc aacgggaatc ctgctctgcg aggctggccg gctaccgccg
     2881 gcgtaacaga tgagggcaag cggatggctg atgaaaccaa gccaaccagg aagggcagcc
     2941 cacctatcaa ggtgtactgc cttccagacg aacgaagagc gattgaggaa aaggcggcgg
     3001 cggccggcat gagcctgtcg gcctacctgc tggccgtcgg ccagggctac aaaatcacgg
     3061 gcgtcgtgga ctatgagcac gtccgcgagc tggccctgca ggcggaagtc aggtagacac
     3121 gacttccgct agtccttgca aggtctgctg acgtgaggcg gggcggtcgt ttttgaccgc
     3181 cctgccttcg tcatgtaggc tcgctcgctg tgcctggcgt gtcatcagac gcccaggtcc
     3241 cggtgccgtg aggcccgggc catcgagccg gtggtacgtg gctgcggtcc ccttgtgagg
     3301 gctgcgcgcc gtgtgctgtc cggcgcgcac agccttgaat ccacccgcgg gggccggccg
     3361 gtctccgtga gctcgagtag acgacggaga cgtaatggac aagaagtaca gcatcggcct
     3421 ggacatcggc accaacagcg tgggctgggc ggtcatcacc gacgagtaca aggtcccctc
     3481 caagaagttc aaggtcctgg gcaacaccga ccggcactcg atcaagaaga acctgatcgg
     3541 cgccctgctc ttcgacagcg gcgaaaccgc cgaggcgacc cgcctgaagc ggaccgcccg
     3601 tcgccgctac acccggcgca agaaccgcat ctgctacctg caggagatct tctccaacga
     3661 gatggccaag gtcgacgact cgttcttcca ccggctcgag gagagcttcc tggtggagga
     3721 ggacaagaag cacgagcgcc acccgatctt cggcaacatc gtcgacgagg tggcctacca
     3781 cgagaagtac cccaccatct accacctccg caagaagctg gtcgactcga ccgacaaggc
     3841 ggacctgcgg ctcatctacc tggccctcgc gcacatgatc aagttccgcg gccacttcct
     3901 catcgagggc gacctgaacc cggacaactc cgacgtcgac aagctcttca tccagctggt
     3961 gcagacctac aaccagctgt tcgaggagaa ccccatcaac gccagcggcg tcgacgccaa
     4021 ggcgatcctc tccgcgcgcc tgagcaagtc ccggcgcctg gagaacctca tcgcccagct
     4081 gccgggcgag aagaagaacg gcctcttcgg caacctgatc gcgctgtcgc tcggcctgac
     4141 ccccaacttc aagagcaact tcgacctggc cgaggacgcg aagctccagc tgtccaagga
     4201 cacctacgac gacgacctgg acaacctgct cgcccagatc ggcgaccagt acgcggacct
     4261 cttcctggcc gcgaagaacc tctcggacgc catcctgctc agcgacatcc tgcgggtcaa
     4321 caccgagatc accaaggccc cgctgtcggc gagcatgatc aagcggtacg acgagcacca
     4381 ccaggacctg accctgctca aggccctcgt gcgccagcag ctgcccgaga agtacaagga
     4441 gatcttcttc gaccagtcca agaacggcta cgccggctac atcgacggcg gcgcgtcgca
     4501 ggaggagttc tacaagttca tcaagccgat cctggagaag atggacggca ccgaggagct
     4561 gctcgtcaag ctgaaccgcg aggacctgct ccgcaagcag cggaccttcg acaacggctc
     4621 catcccgcac cagatccacc tgggcgagct ccacgccatc ctccggcgcc aggaggactt
     4681 ctaccccttc ctgaaggaca accgcgagaa gatcgagaag atcctgacct tccggatccc
     4741 gtactacgtc ggccccctgg cccgcggcaa ctcccggttc gcgtggatga cccggaagtc
     4801 ggaggaaacc atcaccccgt ggaacttcga ggaggtcgtg gacaagggcg cctccgcgca
     4861 gtcgttcatc gagcgcatga ccaacttcga caagaacctc ccgaacgaga aggtcctgcc
     4921 caagcacagc ctgctctacg agtacttcac cgtgtacaac gagctgacca aggtcaagta
     4981 cgtgaccgag ggcatgcgga agccggcctt cctgtccggc gagcagaaga aggcgatcgt
     5041 cgacctgctc ttcaagacca accgcaaggt caccgtgaag cagctgaagg aggactactt
     5101 caagaagatc gagtgcttcg actccgtcga gatctcgggc gtggaggacc gcttcaacgc
     5161 ctccctgggc acctaccacg acctgctcaa gatcatcaag gacaaggact tcctcgacaa
     5221 cgaggagaac gaggacatcc tggaggacat cgtcctcacc ctgaccctct tcgaggaccg
     5281 cgagatgatc gaggagcggc tcaagaccta cgcccacctg ttcgacgaca aggtgatgaa
     5341 gcagctgaag cggcgccggt acaccggctg gggccgcctc tcccggaagc tgatcaacgg
     5401 catccgggac aagcagagcg gcaagaccat cctggacttc ctcaagtccg acggcttcgc
     5461 caaccgcaac ttcatgcagc tcatccacga cgactcgctg accttcaagg aggacatcca
     5521 gaaggcccag gtgtccggcc agggcgacag cctccacgag cacatcgcca acctggcggg
     5581 ctccccggcg atcaagaagg gcatcctcca gaccgtcaag gtcgtggacg agctggtcaa
     5641 ggtgatgggc cgccacaagc ccgagaacat cgtgatcgag atggcccggg agaaccagac
     5701 cacccagaag ggccagaaga actcccgcga gcggatgaag cgcatcgagg agggcatcaa
     5761 ggagctcggc tcgcagatcc tgaaggagca cccggtcgag aacacccagc tccagaacga
     5821 gaagctgtac ctctactacc tgcagaacgg ccgcgacatg tacgtggacc aggagctcga
     5881 catcaaccgg ctgagcgact acgacgtcga ccacatcgtg ccgcagtcct tcctgaagga
     5941 cgactcgatc gacaacaagg tcctgacccg ctccgacaag aaccggggca agtccgacaa
     6001 cgtgccctcg gaggaggtcg tgaagaagat gaagaactac tggcgccagc tgctcaacgc
     6061 caagctcatc acccagcgca agttcgacaa cctgaccaag gccgagcggg gcggcctgtc
     6121 ggagctcgac aaggcgggct tcatcaagcg ccagctcgtc gaaacccggc agatcaccaa
     6181 gcacgtggcc cagatcctgg acagccggat gaacaccaag tacgacgaga acgacaagct
     6241 gatccgcgag gtcaaggtga tcaccctcaa gagcaagctg gtgtccgact tccgcaagga
     6301 cttccagttc tacaaggtcc gggagatcaa caactaccac cacgcccacg acgcgtacct
     6361 gaacgccgtc gtgggcaccg cgctgatcaa gaagtacccg aagctggagt ccgagttcgt
     6421 ctacggcgac tacaaggtct acgacgtgcg caagatgatc gccaagtcgg agcaggagat
     6481 cggcaaggcc accgcgaagt acttcttcta cagcaacatc atgaacttct tcaagaccga
     6541 gatcaccctg gccaacggcg agatccgcaa gcggcccctg atcgaaacca acggcgaaac
     6601 cggcgagatc gtctgggaca agggccgcga cttcgccacc gtccggaagg tgctgtccat
     6661 gccgcaggtc aacatcgtca agaaaaccga ggtgcagacc ggcggcttca gcaaggagtc
     6721 catcctcccc aagcgcaact cggacaagct gatcgcccgg aagaaggact gggacccgaa
     6781 gaagtacggc ggcttcgaca gccccaccgt cgcctactcc gtgctggtcg tggcgaaggt
     6841 cgagaagggc aagagcaaga agctgaagtc cgtgaaggag ctgctcggca tcaccatcat
     6901 ggagcgctcc tcgttcgaga agaacccgat cgacttcctg gaggccaagg gctacaagga
     6961 ggtcaagaag gacctcatca tcaagctgcc caagtactcg ctgttcgagc tcgagaacgg
     7021 ccgcaagcgg atgctcgcca gcgcgggcga gctgcagaag ggcaacgagc tggccctccc
     7081 gtccaagtac gtcaacttcc tgtacctcgc gtcccactac gagaagctga agggctcgcc
     7141 cgaggacaac gagcagaagc agctcttcgt ggagcagcac aagcactacc tggacgagat
     7201 catcgagcag atctcggagt tcagcaagcg ggtcatcctg gccgacgcga acctcgacaa
     7261 ggtgctgtcc gcctacaaca agcaccgcga caagccgatc cgggagcagg cggagaacat
     7321 catccacctg ttcaccctca ccaacctggg cgcccccgcc gcgttcaagt acttcgacac
     7381 caccatcgac cgcaagcggt acaccagcac caaggaggtc ctcgacgcga ccctgatcca
     7441 ccagtccatc accggcctgt acgaaacccg catcgacctc tcccagctcg gcggcgactg
     7501 agaattcaga tctacgcgtt ccccgcaaaa gcggcctttg actccctgca agcctcagcg
     7561 accgaatata tcggttatgc gtgggcgatg gttgttgtca ttgtcggcgc aactatcggt
     7621 atcaagctgt ttaagaaatt cacctcgaaa gcaagctgat aaaccgatac aattaaaggc
     7681 tccttttgga gccttttttt gctgctcctt cggtcggacg tgcgtctacg ggcaccttac
     7741 cgcagccgtc ggctgtgcga cacggacgga tcgggcgaac tggccgatgc tgggagaagc
     7801 gcgctgctgt acggcgcgca ccgggtgcgg agcccctcgg cgagcggtgt gaaacttctg
     7861 tgaatggcct gttcggttgc tttttttata cggctgccag ataaggcttg cagcatctgg
     7921 gcggctaccg ctatgatcgg ggcgttcctg caattcttag tgcgagtatc tgaaagggga
     7981 tacgcaagtc ttctcagccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact
     8041 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
     8101 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
     8161 cgacggccag tgagcgcgcg taatacgact cactataggg cgaattgggt accgggcccc
     8221 ccctcgaggt cctccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa
     8281 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     8341 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     8401 attgcgttgc gctcactgcc cgctttccac cggtgaagac atgttttaga gctagaaata
     8461 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
     8521 tttactccat ctggatttgt tcagaacgct cggttgccgc cgggcgtttt ttatctagag
     8581 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     8641 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     8701 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     8761 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     8821 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgacccc
     8881 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     8941 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     9001 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     9061 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     9121 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     9181 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     9241 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     9301 acctagatcc ttttggttca tgtgcagctc catcagcaaa aggggatgat aagtttatca
     9361 ccaccgacta tttgcaacag tgccgttgat cgtgctatga tcgactgatg tcatcagcgg
     9421 tggagtgcaa tgtcgtgcaa tacgaatggc gaaaagccga gctcatcggt cagcttctca
     9481 accttggggt tacccccggc ggtgtgctgc tggtccacag ctccttccgt agcgtccggc
     9541 ccctcgaaga tgggccactt ggactgatcg aggccctgcg tgctgcgctg ggtccgggag
     9601 ggacgctcgt catgccctcg tggtcaggtc tggacgacga gccgttcgat cctgccacgt
     9661 cgcccgttac accggacctt ggagttgtct ctgacacatt ctggcgcctg ccaaatgtaa
     9721 agcgcagcgc ccatccattt gcctttgcgg cagcggggcc acaggcagag cagatcatct
     9781 ctgatccatt gcccctgcca cctcactcgc ctgcaagccc ggtcgcccgt gtccatgaac
     9841 tcgatgggca ggtacttctc ctcggcgtgg gacacgatgc caacacgacg ctgcatcttg
     9901 ccgagttgat ggcaaaggtt ccctatgggg tgccgagaca ctgcaccatt cttcaggatg
     9961 gcaagttggt acgcgtcgat tatctcgaga atgaccactg ctgtgagcgc tttgccttgg
    10021 cggacaggtg gctcaaggag aagagccttc agaaggaagg tccagtcggt catgcctttg
    10081 ctcggttgat ccgctcccgc gacattgtgg cgacagccct gggtcaactg ggccgagatc
    10141 cgttgatctt cctgcatccg ccagaggcgg gatgcgaaga atgcgatgcc gctcgccagt
    10201 cgattggctg agctcatgag cggagaacga gatgacgttg gaggggcaag gtcgcgctga
    10261 ttgctggggc aacacgtgga gcggatcggg gattgtcttt cttcagctcg ctgatgatat
    10321 gctgacgctc aatgccgttt ggcctccgac taacgaaaat cccgcatttg gacggctgat
    10381 ccgattggca cggcggacgg cgaatggcgg agcagacgct cgtccggggg caatgagata
    10441 tgaaaaagcc tgaactcacc gcgacgtatc gatgtcgagg ttcctcaggg gagccacccc
    10501 agagaagccc tcggagctga gcggagctat ttccaaagcc atgccagcta gagacagtgc
    10561 acaccgccaa gcatctgcaa aaccctcgcc tggagggaaa gtgcaatgta cgtacgctgg
    10621 tttccctcca gaaagggatt ctgcggcttc tcaactttgg aagaagaggc ggaacgaact
    10681 gctgtttcag cctactcatg tgaggaggct ggtcctttac cctgatgccc gagggcatca
    10741 gggtaaagga ccagcctcgc caactaggag cgctaccaag gcgaacacaa cccaggactt
    10801 ggtagaaccc ttcgccggag aagttcagac tcatcggcat gagcacccct cgccgcgcac
    10861 tcggcaccgg tccgtcgacc accaccagga cgtcgttgtc gacgtcggcc ccgcggctcc
    10921 tgcccgccga acgcgtcgtc gtcgacggcc tggtgctcat cgacgagcac ccggagccag
    10981 gtgaaaagcg ccgg
//
