LOCUS       sequence-434957-       11061 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  an ermE*p-cas9-driven expression, as an alternative plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11061)
  AUTHORS   Ye S, Enghiad B, Zhao H, Takano E
  TITLE     Fine-tuning the regulation of Cas9 expression levels for efficient 
            CRISPR-Cas9 mediated recombination in Streptomyces.
  JOURNAL   J Ind Microbiol Biotechnol. 2020 May;47(4-5):413-423. doi: 
            10.1007/s10295-020-02277-5. Epub 2020 May 4.
  PUBMED    32367443
REFERENCE   2  (bases 1 to 11061)
  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: "J Ind 
            Microbiol Biotechnol."; date: "2020-05"; volume: "47(4-5):413-423. 
            doi"; pages: " 10.1007/s10295-020-02277-5. Epub 2020 May 4"
COMMENT     Sequence Label: pCM4.4
FEATURES             Location/Qualifiers
     source          1..11061
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             172..4278
                     /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             172..4275
                     /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        4478..4762
                     /label=gapdh promoter
                     /note="glyceraldehyde-3-phosphate dehydrogenase promoter 
                     from Eggerthella lenta"
     CDS             complement(4771..5067)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSG"
     primer_bind     4917..4939
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4931..4948
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4932..4948
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4958..4977
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4958..4976
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5024..5044)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5024..5042)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5063..5079)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5063..5079)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5076..5098)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5087..5103
                     /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(5111..5117,5118..5135,5136..5141))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5156..5177
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_RNA        5220..5295
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      5317..5351
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     rep_origin      complement(5393..5981)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5473..5492)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             6185..6985
                     /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"
     CDS             complement(7924..9366)
                     /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            10406..10515
                     /note="incP origin of transfer"
     CDS             10548..10919
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
ORIGIN
        1 gctagcaatt aaaggctcct tttggagcct ttttttacgc ggtcgatctt gacggctggc
       61 gagaggtgcg gggaggatct gaccgacgcg gtccacacgt ggcaccgcga tgctgttgtg
      121 ggcacaatcg tgccggttgg taggatccag cggagcaacg gaggtacgga catggacaag
      181 aagtacagca tcggcctgga catcggcacc aacagcgtgg gctgggcggt catcaccgac
      241 gagtacaagg tcccctccaa gaagttcaag gtcctgggca acaccgaccg gcactcgatc
      301 aagaagaacc tgatcggcgc cctgctcttc gacagcggcg aaaccgccga ggcgacccgc
      361 ctgaagcgga ccgcccgtcg ccgctacacc cggcgcaaga accgcatctg ctacctgcag
      421 gagatcttct ccaacgagat ggccaaggtc gacgactcgt tcttccaccg gctcgaggag
      481 agcttcctgg tggaggagga caagaagcac gagcgccacc cgatcttcgg caacatcgtc
      541 gacgaggtgg cctaccacga gaagtacccc accatctacc acctccgcaa gaagctggtc
      601 gactcgaccg acaaggcgga cctgcggctc atctacctgg ccctcgcgca catgatcaag
      661 ttccgcggcc acttcctcat cgagggcgac ctgaacccgg acaactccga cgtcgacaag
      721 ctcttcatcc agctggtgca gacctacaac cagctgttcg aggagaaccc catcaacgcc
      781 agcggcgtcg acgccaaggc gatcctctcc gcgcgcctga gcaagtcccg gcgcctggag
      841 aacctcatcg cccagctgcc gggcgagaag aagaacggcc tcttcggcaa cctgatcgcg
      901 ctgtcgctcg gcctgacccc caacttcaag agcaacttcg acctggccga ggacgcgaag
      961 ctccagctgt ccaaggacac ctacgacgac gacctggaca acctgctcgc ccagatcggc
     1021 gaccagtacg cggacctctt cctggccgcg aagaacctct cggacgccat cctgctcagc
     1081 gacatcctgc gggtcaacac cgagatcacc aaggccccgc tgtcggcgag catgatcaag
     1141 cggtacgacg agcaccacca ggacctgacc ctgctcaagg ccctcgtgcg ccagcagctg
     1201 cccgagaagt acaaggagat cttcttcgac cagtccaaga acggctacgc cggctacatc
     1261 gacggcggcg cgtcgcagga ggagttctac aagttcatca agccgatcct ggagaagatg
     1321 gacggcaccg aggagctgct cgtcaagctg aaccgcgagg acctgctccg caagcagcgg
     1381 accttcgaca acggctccat cccgcaccag atccacctgg gcgagctcca cgccatcctc
     1441 cggcgccagg aggacttcta ccccttcctg aaggacaacc gcgagaagat cgagaagatc
     1501 ctgaccttcc ggatcccgta ctacgtcggc cccctggccc gcggcaactc ccggttcgcg
     1561 tggatgaccc ggaagtcgga ggaaaccatc accccgtgga acttcgagga ggtcgtggac
     1621 aagggcgcct ccgcgcagtc gttcatcgag cgcatgacca acttcgacaa gaacctcccg
     1681 aacgagaagg tcctgcccaa gcacagcctg ctctacgagt acttcaccgt gtacaacgag
     1741 ctgaccaagg tcaagtacgt gaccgagggc atgcggaagc cggccttcct gtccggcgag
     1801 cagaagaagg cgatcgtcga cctgctcttc aagaccaacc gcaaggtcac cgtgaagcag
     1861 ctgaaggagg actacttcaa gaagatcgag tgcttcgact ccgtcgagat ctcgggcgtg
     1921 gaggaccgct tcaacgcctc cctgggcacc taccacgacc tgctcaagat catcaaggac
     1981 aaggacttcc tcgacaacga ggagaacgag gacatcctgg aggacatcgt cctcaccctg
     2041 accctcttcg aggaccgcga gatgatcgag gagcggctca agacctacgc ccacctgttc
     2101 gacgacaagg tgatgaagca gctgaagcgg cgccggtaca ccggctgggg ccgcctctcc
     2161 cggaagctga tcaacggcat ccgggacaag cagagcggca agaccatcct ggacttcctc
     2221 aagtccgacg gcttcgccaa ccgcaacttc atgcagctca tccacgacga ctcgctgacc
     2281 ttcaaggagg acatccagaa ggcccaggtg tccggccagg gcgacagcct ccacgagcac
     2341 atcgccaacc tggcgggctc cccggcgatc aagaagggca tcctccagac cgtcaaggtc
     2401 gtggacgagc tggtcaaggt gatgggccgc cacaagcccg agaacatcgt gatcgagatg
     2461 gcccgggaga accagaccac ccagaagggc cagaagaact cccgcgagcg gatgaagcgc
     2521 atcgaggagg gcatcaagga gctcggctcg cagatcctga aggagcaccc ggtcgagaac
     2581 acccagctcc agaacgagaa gctgtacctc tactacctgc agaacggccg cgacatgtac
     2641 gtggaccagg agctcgacat caaccggctg agcgactacg acgtcgacca catcgtgccg
     2701 cagtccttcc tgaaggacga ctcgatcgac aacaaggtcc tgacccgctc cgacaagaac
     2761 cggggcaagt ccgacaacgt gccctcggag gaggtcgtga agaagatgaa gaactactgg
     2821 cgccagctgc tcaacgccaa gctcatcacc cagcgcaagt tcgacaacct gaccaaggcc
     2881 gagcggggcg gcctgtcgga gctcgacaag gcgggcttca tcaagcgcca gctcgtcgaa
     2941 acccggcaga tcaccaagca cgtggcccag atcctggaca gccggatgaa caccaagtac
     3001 gacgagaacg acaagctgat ccgcgaggtc aaggtgatca ccctcaagag caagctggtg
     3061 tccgacttcc gcaaggactt ccagttctac aaggtccggg agatcaacaa ctaccaccac
     3121 gcccacgacg cgtacctgaa cgccgtcgtg ggcaccgcgc tgatcaagaa gtacccgaag
     3181 ctggagtccg agttcgtcta cggcgactac aaggtctacg acgtgcgcaa gatgatcgcc
     3241 aagtcggagc aggagatcgg caaggccacc gcgaagtact tcttctacag caacatcatg
     3301 aacttcttca agaccgagat caccctggcc aacggcgaga tccgcaagcg gcccctgatc
     3361 gaaaccaacg gcgaaaccgg cgagatcgtc tgggacaagg gccgcgactt cgccaccgtc
     3421 cggaaggtgc tgtccatgcc gcaggtcaac atcgtcaaga aaaccgaggt gcagaccggc
     3481 ggcttcagca aggagtccat cctccccaag cgcaactcgg acaagctgat cgcccggaag
     3541 aaggactggg acccgaagaa gtacggcggc ttcgacagcc ccaccgtcgc ctactccgtg
     3601 ctggtcgtgg cgaaggtcga gaagggcaag agcaagaagc tgaagtccgt gaaggagctg
     3661 ctcggcatca ccatcatgga gcgctcctcg ttcgagaaga acccgatcga cttcctggag
     3721 gccaagggct acaaggaggt caagaaggac ctcatcatca agctgcccaa gtactcgctg
     3781 ttcgagctcg agaacggccg caagcggatg ctcgccagcg cgggcgagct gcagaagggc
     3841 aacgagctgg ccctcccgtc caagtacgtc aacttcctgt acctcgcgtc ccactacgag
     3901 aagctgaagg gctcgcccga ggacaacgag cagaagcagc tcttcgtgga gcagcacaag
     3961 cactacctgg acgagatcat cgagcagatc tcggagttca gcaagcgggt catcctggcc
     4021 gacgcgaacc tcgacaaggt gctgtccgcc tacaacaagc accgcgacaa gccgatccgg
     4081 gagcaggcgg agaacatcat ccacctgttc accctcacca acctgggcgc ccccgccgcg
     4141 ttcaagtact tcgacaccac catcgaccgc aagcggtaca ccagcaccaa ggaggtcctc
     4201 gacgcgaccc tgatccacca gtccatcacc ggcctgtacg aaacccgcat cgacctctcc
     4261 cagctcggcg gcgactgaga attcagatct acgcgttccc cgcaaaagcg gcctttgact
     4321 ccctgcaagc ctcagcgacc gaatatatcg gttatgcgtg ggcgatggtt gttgtcattg
     4381 tcggcgcaac tatcggtatc aagctgttta agaaattcac ctcgaaagca agctgataaa
     4441 ccgatacaat taaaggctcc ttttggagcc tttttttgct gctccttcgg tcggacgtgc
     4501 gtctacgggc accttaccgc agccgtcggc tgtgcgacac ggacggatcg ggcgaactgg
     4561 ccgatgctgg gagaagcgcg ctgctgtacg gcgcgcaccg ggtgcggagc ccctcggcga
     4621 gcggtgtgaa acttctgtga atggcctgtt cggttgcttt ttttatacgg ctgccagata
     4681 aggcttgcag catctgggcg gctaccgcta tgatcggggc gttcctgcaa ttcttagtgc
     4741 gagtatctga aaggggatac gcaagtcttc tcagccgcta cagggcgcgt cccattcgcc
     4801 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     4861 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     4921 gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac tatagggcga
     4981 attgggtacc gggccccccc tcgaggtcct ccagcttttg ttccctttag tgagggttaa
     5041 ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5101 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5161 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccaccgg tgaagacatg
     5221 ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg
     5281 gcaccgagtc ggtgcttttt actccatctg gatttgttca gaacgctcgg ttgccgccgg
     5341 gcgtttttta tctagaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5401 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5461 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5521 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5581 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5641 gctgtgtgca cgaccccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5701 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5761 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5821 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5881 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5941 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6001 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     6061 tatcaaaaag gatcttcacc tagatccttt tggttcatgt gcagctccat cagcaaaagg
     6121 ggatgataag tttatcacca ccgactattt gcaacagtgc cgttgatcgt gctatgatcg
     6181 actgatgtca tcagcggtgg agtgcaatgt cgtgcaatac gaatggcgaa aagccgagct
     6241 catcggtcag cttctcaacc ttggggttac ccccggcggt gtgctgctgg tccacagctc
     6301 cttccgtagc gtccggcccc tcgaagatgg gccacttgga ctgatcgagg ccctgcgtgc
     6361 tgcgctgggt ccgggaggga cgctcgtcat gccctcgtgg tcaggtctgg acgacgagcc
     6421 gttcgatcct gccacgtcgc ccgttacacc ggaccttgga gttgtctctg acacattctg
     6481 gcgcctgcca aatgtaaagc gcagcgccca tccatttgcc tttgcggcag cggggccaca
     6541 ggcagagcag atcatctctg atccattgcc cctgccacct cactcgcctg caagcccggt
     6601 cgcccgtgtc catgaactcg atgggcaggt acttctcctc ggcgtgggac acgatgccaa
     6661 cacgacgctg catcttgccg agttgatggc aaaggttccc tatggggtgc cgagacactg
     6721 caccattctt caggatggca agttggtacg cgtcgattat ctcgagaatg accactgctg
     6781 tgagcgcttt gccttggcgg acaggtggct caaggagaag agccttcaga aggaaggtcc
     6841 agtcggtcat gcctttgctc ggttgatccg ctcccgcgac attgtggcga cagccctggg
     6901 tcaactgggc cgagatccgt tgatcttcct gcatccgcca gaggcgggat gcgaagaatg
     6961 cgatgccgct cgccagtcga ttggctgagc tcatgagcgg agaacgagat gacgttggag
     7021 gggcaaggtc gcgctgattg ctggggcaac acgtggagcg gatcggggat tgtctttctt
     7081 cagctcgctg atgatatgct gacgctcaat gccgtttggc ctccgactaa cgaaaatccc
     7141 gcatttggac ggctgatccg attggcacgg cggacggcga atggcggagc agacgctcgt
     7201 ccgggggcaa tgagatatga aaaagcctga actcaccgcg acgtatcgat gtcgaggttc
     7261 ctcaggggag ccaccccaga gaagccctcg gagctgagcg gagctatttc caaagccatg
     7321 ccagctagag acagtgcaca ccgccaagca tctgcaaaac cctcgcctgg agggaaagtg
     7381 caatgtacgt acgctggttt ccctccagaa agggattctg cggcttctca actttggaag
     7441 aagaggcgga acgaactgct gtttcagcct actcatgtga ggaggctggt cctttaccct
     7501 gatgcccgag ggcatcaggg taaaggacca gcctcgccaa ctaggagcgc taccaaggcg
     7561 aacacaaccc aggacttggt agaacccttc gccggagaag ttcagactca tcggcatgag
     7621 cacccctcgc cgcgcactcg gcaccggtcc gtcgaccacc accaggacgt cgttgtcgac
     7681 gtcggccccg cggctcctgc ccgccgaacg cgtcgtcgtc gacggcctgg tgctcatcga
     7741 cgagcacccg gagccaggtg aaaagcgccg gcggacgctc ggactgggcg cgggattcca
     7801 gcagtaaccc aggtccgccg gccacctcac ggcaggcaga ccctcggctt tcgcgccggg
     7861 accgccatga gaccgccacc cggatgtccg gggtggcggt ctcatggcgg tccctcagcg
     7921 gccctacgcg accgctgtgt cgacgcggag gcagtctccg gggtgctggc cccagggctg
     7981 gagtaccggg gcgagcttgc ccttgcagcg gcggcacacg ggcgacgcgg cgccgggcgg
     8041 gggagtcttg acgtcgacgg tgaccggctt cggcttcagg cgcttccgaa ggctgatcgt
     8101 cgggcggatc gcctccttct tcggctgacg cactcggtcg agcgaagcgg ccagctcctc
     8161 cgtgcgggcc tcgttggcct tccggcgcgc ctcgcggaaa gcagcttcct cctcggacat
     8221 gacctcgaac ctcatctggt cagcgtcaag gtcgcccggc gcggccggcg cttccggggc
     8281 gggcgggtcc aggacgtcct tgccccacac caggccccag gactcgacga gccgccggac
     8341 gcccggtagg ccgtacgtct cggcgacctt gatgaggtcg aggcgacgtc cggcgacgcg
     8401 ggcgatgtat cggtaccaga tgtaggccgg gatgaccgcg atggcgacca ggccctcggt
     8461 gtcgtcggtg atctcctcct cggtgcggac gtcctgctgg atgccgagtt ccttgatcag
     8521 ccggttcagg ttctgcgacc ggtagtgctt gcggacctgg aacacgccga actcgcgctc
     8581 gcggtacttc tcgacgaacg ggccgggccg acgaagccgc tgcagctcgg cggccgccgc
     8641 gtcgcccagg tcgagcggtc ccatgcggtc gtcgccgcga ccggccttga agttctgtcc
     8701 ggccagctcc aggccgatct tggcgacgcc gcccttggtc ttgtcgccgt ccttgtagag
     8761 gtagcgggcc tgcttgcccg catcgccgtc agcggcgtcc gcgccgttga gtgggcgcac
     8821 gtcggtgccg tggcccttgc cctcacagga gcaaccgggc cggtcgcacg tctcgctgac
     8881 ggtgtagccg cccgcggatt cgaccccggc ggcccaggct ccggcgagtg cgtcgcggaa
     8941 cgcggcctgg gcgtccgggc cgagcacctc gcgggtgacc cagagcgtgt gccagtgcag
     9001 gtgccagccg gagccccagc cgaaggtgtc ctcgaaggcc cgctcgtagc cgatgatccc
     9061 gaagtcgtcg cgcatcgtgc gccagcggcg gccggacgag ccgtacgcgc ccttccagcc
     9121 gtcgtgcaag accgcgacca ggccgtgccg cattcccttg cggacggtgc cgaacgccat
     9181 gcgctcgaag tggcgcaacg tgttcgtgcc aaggtgcagc ccgtacccgg cgtccgcgag
     9241 accgtcggcg gcgagctgca cgttcgagcc ccgtacggcc aggatgcggc tcatgcacca
     9301 cgggcaggtg tggacgttgt tgcagcggca cgtgttgccc cacgtcgcct cgcccggctt
     9361 ccacatcagc tcggccgtcc cggcagtgag ccgggtcccg cagcccttga acgcctcgtt
     9421 cagcgacacc gtctggtgcc ggtcccgccg ggcgaaccgc tcgtcgcgcg ggtcctcccg
     9481 cccggctgtc gcggcaccct cgtttggggt agaacccgtt ccagttacag cgctctgacc
     9541 tgcagtggac ggagattttc ccttactact aaagcccgcg tccggattac ccgctgtagt
     9601 cgtgcttgct acgctgcgtg actggtccgc aatgagacgc tttgcgcgct ttcggcaggc
     9661 gtccgagcag tagattttgg ggcgcttccc ggggatgtgg acgatcgggg tgccgcagtg
     9721 gcacttcggt ccggcggggc gcggtggtgt cgacgcgctg ttctctcgta cgctcgtcac
     9781 agagcaaacg tcctcactcg gcatgctgcg ccggttcggg ggcggcgagc ccgggaggcc
     9841 aatcccgggc tcgtgccatt tctgggtcct gttgatcctg gcattggtgt ggccgttcat
     9901 tgcccctgct cgctcctgac gcgccgatag acgtccgata cgcccggtgc tggtgggatt
     9961 tgataggtcg gaagaagccc cgcccgggcc tgggcggggc ttcctgtgcg tcaggacctc
    10021 ctcgtcgtga gcctcttcgg cctatggacg gagtgacctc gtgatccgtt acagccgcgc
    10081 gcgctcgcgt agagcggtct catcagttcc acgaacggtc ctcttcgcag atcagggcgt
    10141 tggggcggag tctcaccaag gactacgtct gctggcgatt tccgttacac cccgggcggt
    10201 ggccggcgca cacgcgcgcc cgcgttgggc agtgcagaaa gtgcagaaac ctaggtgcag
    10261 gtcgacggat cttttccgct gcataaccct gcttcggggt cattatagcg attttttcgg
    10321 tatatccatc ctttttcgca cgatatacag gattttgcca aagggttcgt gtagactttc
    10381 cttggtgtat ccaacggcgt cagccgggca ggataggtga agtaggccca cccgcgagcg
    10441 ggtgttcctt cttcactgtc ccttattcgc acctggcggt gctcaacggg aatcctgctc
    10501 tgcgaggctg gccggctacc gccggcgtaa cagatgaggg caagcggatg gctgatgaaa
    10561 ccaagccaac caggaagggc agcccaccta tcaaggtgta ctgccttcca gacgaacgaa
    10621 gagcgattga ggaaaaggcg gcggcggccg gcatgagcct gtcggcctac ctgctggccg
    10681 tcggccaggg ctacaaaatc acgggcgtcg tggactatga gcacgtccgc gagctggccc
    10741 gcatcaatgg cgacctgggc cgcctgggcg gcctgctgaa actctggctc accgacgacc
    10801 cgcgcacggc gcggttcggt gatgccacga tcctcgccct gctggcgaag atcgaagaga
    10861 agcaggacga gcttggcaag gtcatgatgg gcgtggtccg cccgagggca gagccatgac
    10921 ttttttagcc gctaaaacgg ccggggggtg cgcgtgattg ccaagcacgt ccccatgcgc
    10981 tccatcaaga agagcgactt cgcggagctg gtgaagtaca tcaccgacga gcaaggcaag
    11041 accgatcccc ggggacctgc a
//
