LOCUS       sequence-129198-       10841 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of Cas9 gene and inducible expression of the
            plasmid curing system for CRISPR mediated genome editing of E. 
            coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10841)
  AUTHORS   Li Y, Lin Z, Huang C, Zhang Y, Wang Z, Tang YJ, Chen T, Zhao X
  TITLE     Metabolic engineering of Escherichia coli using CRISPR-Cas9 
            meditated genome editing.
  JOURNAL   Metab Eng. 2015 Sep;31:13-21. doi: 10.1016/j.ymben.2015.06.006. Epub
            2015 Jun 30.
  PUBMED    26141150
REFERENCE   2  (bases 1 to 10841)
  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: "Metab 
            Eng."; date: "2015-09"; volume: "31:13-21. doi"; pages: " 
            10.1016/j.ymben.2015.06.006. Epub 2015 Jun 30"
COMMENT     Sequence Label: pCas9cur
FEATURES             Location/Qualifiers
     source          1..10841
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     134..153
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(220..322)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(714..731)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(848..1393)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(928..947)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(1505..1510,1511..1527,1528..1533)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     misc_RNA        complement(1851..1929)
                     /label=tracrRNA
                     /note="trans-activating CRISPR RNA for the Streptococcus 
                     pyogenes CRISPR/Cas9 system"
     CDS             2225..6331
                     /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             2225..6328
                     /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"
     misc_feature    6352..6483
                     /label=crRNA leader
                     /note="crRNA leader sequence for the Streptococcus pyogenes
                     CRISPR/Cas system"
     repeat_region   6484..6519
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     CDS             complement(6729..7607)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        7634..7918
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     7841..7860
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     repeat_region   8065..8100
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     primer_bind     complement(8529..8548)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(join(10401..10841,1..219))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 gaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt
       61 gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa cggtctggtt
      121 ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat gccattggga
      181 tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct tagctcctga
      241 aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt
      301 ggaacctctt acgtgccgat caacgtctca ttttcgccaa aagttggccc agggcttccc
      361 ggtatcaaca gggacaccag gatttattta ttctgcgaag tgatcttccg tcacaggtat
      421 ttattcggcg caaagtgcgt cgggtgatgc tgccaactta ctgatttagt gtatgatggt
      481 gtttttgagg tgctccagtg gcttctgttt ctatcagctg tccctcctgt tcagctactg
      541 acggggtggt gcgtaacggc aaaagcaccg ccggacatca gcgctagcgg agtgtatact
      601 ggcttactat gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa
      661 aaggctgcac cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc
      721 actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc
      781 ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa
      841 agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc
      901 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc
      961 tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc
     1021 gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac
     1081 tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt
     1141 gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt
     1201 agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg
     1261 tgactgcgct cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt
     1321 cgaaaaaccg ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc
     1381 aaaacgatct caagaagatc atcttattaa tcagataaaa tatttctaga tttcagtgca
     1441 atttatctct tcaaatgtag cacctgaagt cagccccata cgatataagt tgtaattctc
     1501 atgtttgaca gcttatcatc gataagcttt aatgcggtag tttatcacag ttaaattgct
     1561 aacgcagtca ggcaccgtgt atgaaatcta acaatgcgct catcgtcatc ctcggcaccg
     1621 tcaccctgga tgctgtaggc ataggcttgg ttatgccggt actgccgggc ctcttgcggg
     1681 attacgaaat catcctgtgg agcttagtag gtttagcaag atggcagcgc ctaaatgtag
     1741 aatgataaaa ggattaagag attaatttcc ctaaaaatga taaaacaagc gttttgaaag
     1801 cgcttgtttt tttggtttgc agtcagagta gaatagaagt atcaaaaaaa gcaccgactc
     1861 ggtgccactt tttcaagttg ataacggact agccttattt taacttgcta tgctgttttg
     1921 aatggttcca acaagattat tttataactt ttataacaaa taatcaagga gaaattcaaa
     1981 gaaatttatc agccataaaa caatacttaa tactatagaa tgataacaaa ataaactact
     2041 ttttaaaaga attttgtgtt ataatctatt tattattaag tattgggtaa tattttttga
     2101 agagatattt tgaaaaagaa aaattaaagc atattaaact aatttcggag gtcattaaaa
     2161 ctattattga aatcatcaaa ctcattatgg atttaattta aactttttat tttaggaggc
     2221 aaaaatggat aagaaatact caataggctt agatatcggc acaaatagcg tcggatgggc
     2281 ggtgatcact gatgaatata aggttccgtc taaaaagttc aaggttctgg gaaatacaga
     2341 ccgccacagt atcaaaaaaa atcttatagg ggctctttta tttgacagtg gagagacagc
     2401 ggaagcgact cgtctcaaac ggacagctcg tagaaggtat acacgtcgga agaatcgtat
     2461 ttgttatcta caggagattt tttcaaatga gatggcgaaa gtagatgata gtttctttca
     2521 tcgacttgaa gagtcttttt tggtggaaga agacaagaag catgaacgtc atcctatttt
     2581 tggaaatata gtagatgaag ttgcttatca tgagaaatat ccaactatct atcatctgcg
     2641 aaaaaaattg gtagattcta ctgataaagc ggatttgcgc ttaatctatt tggccttagc
     2701 gcatatgatt aagtttcgtg gtcatttttt gattgaggga gatttaaatc ctgataatag
     2761 tgatgtggac aaactattta tccagttggt acaaacctac aatcaattat ttgaagaaaa
     2821 ccctattaac gcaagtggag tagatgctaa agcgattctt tctgcacgat tgagtaaatc
     2881 aagacgatta gaaaatctca ttgctcagct ccccggtgag aagaaaaatg gcttatttgg
     2941 gaatctcatt gctttgtcat tgggtttgac ccctaatttt aaatcaaatt ttgatttggc
     3001 agaagatgct aaattacagc tttcaaaaga tacttacgat gatgatttag ataatttatt
     3061 ggcgcaaatt ggagatcaat atgctgattt gtttttggca gctaagaatt tatcagatgc
     3121 tattttactt tcagatatcc taagagtaaa tactgaaata actaaggctc ccctatcagc
     3181 ttcaatgatt aaacgctacg atgaacatca tcaagacttg actcttttaa aagctttagt
     3241 tcgacaacaa cttccagaaa agtataaaga aatctttttt gatcaatcaa aaaacggata
     3301 tgcaggttat attgatgggg gagctagcca agaagaattt tataaattta tcaaaccaat
     3361 tttagaaaaa atggatggta ctgaggaatt attggtgaaa ctaaatcgtg aagatttgct
     3421 gcgcaagcaa cggacctttg acaacggctc tattccccat caaattcact tgggtgagct
     3481 gcatgctatt ttgagaagac aagaagactt ttatccattt ttaaaagaca atcgtgagaa
     3541 gattgaaaaa atcttgactt ttcgaattcc ttattatgtt ggtccattgg cgcgtggcaa
     3601 tagtcgtttt gcatggatga ctcggaagtc tgaagaaaca attaccccat ggaattttga
     3661 agaagttgtc gataaaggtg cttcagctca atcatttatt gaacgcatga caaactttga
     3721 taaaaatctt ccaaatgaaa aagtactacc aaaacatagt ttgctttatg agtattttac
     3781 ggtttataac gaattgacaa aggtcaaata tgttactgaa ggaatgcgaa aaccagcatt
     3841 tctttcaggt gaacagaaga aagccattgt tgatttactc ttcaaaacaa atcgaaaagt
     3901 aaccgttaag caattaaaag aagattattt caaaaaaata gaatgttttg atagtgttga
     3961 aatttcagga gttgaagata gatttaatgc ttcattaggt acctaccatg atttgctaaa
     4021 aattattaaa gataaagatt ttttggataa tgaagaaaat gaagatatct tagaggatat
     4081 tgttttaaca ttgaccttat ttgaagatag ggagatgatt gaggaaagac ttaaaacata
     4141 tgctcacctc tttgatgata aggtgatgaa acagcttaaa cgtcgccgtt atactggttg
     4201 gggacgtttg tctcgaaaat tgattaatgg tattagggat aagcaatctg gcaaaacaat
     4261 attagatttt ttgaaatcag atggttttgc caatcgcaat tttatgcagc tgatccatga
     4321 tgatagtttg acatttaaag aagacattca aaaagcacaa gtgtctggac aaggcgatag
     4381 tttacatgaa catattgcaa atttagctgg tagccctgct attaaaaaag gtattttaca
     4441 gactgtaaaa gttgttgatg aattggtcaa agtaatgggg cggcataagc cagaaaatat
     4501 cgttattgaa atggcacgtg aaaatcagac aactcaaaag ggccagaaaa attcgcgaga
     4561 gcgtatgaaa cgaatcgaag aaggtatcaa agaattagga agtcagattc ttaaagagca
     4621 tcctgttgaa aatactcaat tgcaaaatga aaagctctat ctctattatc tccaaaatgg
     4681 aagagacatg tatgtggacc aagaattaga tattaatcgt ttaagtgatt atgatgtcga
     4741 tcacattgtt ccacaaagtt tccttaaaga cgattcaata gacaataagg tcttaacgcg
     4801 ttctgataaa aatcgtggta aatcggataa cgttccaagt gaagaagtag tcaaaaagat
     4861 gaaaaactat tggagacaac ttctaaacgc caagttaatc actcaacgta agtttgataa
     4921 tttaacgaaa gctgaacgtg gaggtttgag tgaacttgat aaagctggtt ttatcaaacg
     4981 ccaattggtt gaaactcgcc aaatcactaa gcatgtggca caaattttgg atagtcgcat
     5041 gaatactaaa tacgatgaaa atgataaact tattcgagag gttaaagtga ttaccttaaa
     5101 atctaaatta gtttctgact tccgaaaaga tttccaattc tataaagtac gtgagattaa
     5161 caattaccat catgcccatg atgcgtatct aaatgccgtc gttggaactg ctttgattaa
     5221 gaaatatcca aaacttgaat cggagtttgt ctatggtgat tataaagttt atgatgttcg
     5281 taaaatgatt gctaagtctg agcaagaaat aggcaaagca accgcaaaat atttctttta
     5341 ctctaatatc atgaacttct tcaaaacaga aattacactt gcaaatggag agattcgcaa
     5401 acgccctcta atcgaaacta atggggaaac tggagaaatt gtctgggata aagggcgaga
     5461 ttttgccaca gtgcgcaaag tattgtccat gccccaagtc aatattgtca agaaaacaga
     5521 agtacagaca ggcggattct ccaaggagtc aattttacca aaaagaaatt cggacaagct
     5581 tattgctcgt aaaaaagact gggatccaaa aaaatatggt ggttttgata gtccaacggt
     5641 agcttattca gtcctagtgg ttgctaaggt ggaaaaaggg aaatcgaaga agttaaaatc
     5701 cgttaaagag ttactaggga tcacaattat ggaaagaagt tcctttgaaa aaaatccgat
     5761 tgacttttta gaagctaaag gatataagga agttaaaaaa gacttaatca ttaaactacc
     5821 taaatatagt ctttttgagt tagaaaacgg tcgtaaacgg atgctggcta gtgccggaga
     5881 attacaaaaa ggaaatgagc tggctctgcc aagcaaatat gtgaattttt tatatttagc
     5941 tagtcattat gaaaagttga agggtagtcc agaagataac gaacaaaaac aattgtttgt
     6001 ggagcagcat aagcattatt tagatgagat tattgagcaa atcagtgaat tttctaagcg
     6061 tgttatttta gcagatgcca atttagataa agttcttagt gcatataaca aacatagaga
     6121 caaaccaata cgtgaacaag cagaaaatat tattcattta tttacgttga cgaatcttgg
     6181 agctcccgct gcttttaaat attttgatac aacaattgat cgtaaacgat atacgtctac
     6241 aaaagaagtt ttagatgcca ctcttatcca tcaatccatc actggtcttt atgaaacacg
     6301 cattgatttg agtcagctag gaggtgactg aagtatattt tagatgaaga ttatttctta
     6361 ataactaaaa atatggtata atactcttaa taaatgcagt aatacagggg cttttcaaga
     6421 ctgaagtcta gctgagacaa atagtgcgat tacgaaattt tttagacaaa aatagtctac
     6481 gaggttttag agctatgctg ttttgaatgg tcccaaaact gcagcgcaat agttggcgaa
     6541 gtaatcgcaa catccgcatt aaaatctagc gagggcttta ctaagctccg caaaaaaccc
     6601 cgcccctgac agggcggggt tttttcgcgc ccgatccttg gagcccttgc cctcccgcac
     6661 gatgatcgtg ccgtgatcga aatccagatc cttgacccgc agttgcaaac cctcactgat
     6721 ccgcatgctt atgacaactt gacggctaca tcattcactt tttcttcaca accggcacgg
     6781 aactcgctcg ggctggcccc ggtgcatttt ttaaataccc gcgagaaata gagttgatcg
     6841 tcaaaaccaa cattgcgacc gacggtggcg ataggcatcc gggtggtgct caaaagcagc
     6901 ttcgcctggc tgatacgttg gtcctcgcgc cagcttaaga cgctaatccc taactgctgg
     6961 cggaaaagat gtgacagacg cgacggcgac aagcaaacat gctgtgcgac gctggcgata
     7021 tcaaaattgc tgtctgccag gtgatcgctg atgtactgac aagcctcgcg tacccgatta
     7081 tccatcggtg gatggagcga ctcgttaatc gcttccatgc gccgcagtaa caattgctca
     7141 agcagattta tcgccagcag ctccgaatag cgcccttccc cttgcccggc gttaatgatt
     7201 tgcccaaaca ggtcgctgaa atgcggctgg tgcgcttcat ccgggcgaaa gaaccccgta
     7261 ttggcaaata ttgacggcca gttaagccat tcatgccagt aggcgcgcgg acgaaagtaa
     7321 acccactggt gataccattc gcgagcctcc ggatgacgac cgtagtgatg aatctctcct
     7381 ggcgggaaca gcaaaatatc acccggtcgg caaacaaatt ctcgtccctg atttttcacc
     7441 accccctgac cgcgaatggt gagattgaga atataacctt tcattcccag cggtcggtcg
     7501 ataaaaaaat cgagataacc gttggcctca atcggcgtta aacccgccac cagatgggca
     7561 ttaaacgagt atcccggcag caggggatca ttttgcgctt cagccatact tttcatactc
     7621 ccgccattca gagaagaaac caattgtcca tattgcatca gacattgccg tcactgcgtc
     7681 ttttactggc tcttctcgct aaccaaaccg gtaaccccgc ttattaaaag cattctgtaa
     7741 caaagcggga ccaaagccat gacaaaaacg cgtaacaaaa gtgtctataa tcacggcaga
     7801 aaagtccaca ttgattattt gcacggcgtc acactttgct atgccatagc atttttatcc
     7861 ataagattag cggatcctac ctgacgcttt ttatcgcaac tctctactgt ttctccatgt
     7921 aggtcgttcg ctccaagcgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg
     7981 ttatcaactt gaaaaagtgg caccagaccc cgttgatgat accgctgcct tactgggtgc
     8041 attagccagt ctgaatgacc tcgagtttta gagctatgct gttttgaatg gtcccaaaac
     8101 ttcagcacac tgagacttgt tgagttccat gttttagagc tatgctgttt tgaatggact
     8161 ccattcaaca ttgccgatga taacttgaga aagagggtta ataccagcag tcggatacct
     8221 tcctattctt tctgttaaag cgttttcatg ttataatagg caaaagaaga gtagtgtgat
     8281 cgtccattcc gacagcatcg ccagtcacta tggcgtgctg ctagcgctat atgcgttgat
     8341 gcaatttcta tgcgcacccg ttctcggagc actgtccgac cgctttggcc gccgcccagt
     8401 cctgctcgct tcgctacttg gagccactat cgactacgcg atcatggcga ccacacccgt
     8461 cctgtggatc ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt
     8521 tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct
     8581 catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg
     8641 cgccatctcc ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc tcaacctact
     8701 actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgaccga tgcccttgag
     8761 agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact
     8821 tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tctgggtcat
     8881 tttcggcgag gaccgctttc gctggagcgc gacgatgatc ggcctgtcgc ttgcggtatt
     8941 cggaatcttg cacgccctcg ctcaagcctt cgtcactggt cccgccacca aacgtttcgg
     9001 cgagaagcag gccattatcg ccggcatggc ggccgacgcg ctgggctacg tcttgctggc
     9061 gttcgcgacg cgaggctgga tggccttccc cattatgatt cttctcgctt ccggcggcat
     9121 cgggatgccc gcgttgcagg ccatgctgtc caggcaggta gatgacgacc atcagggaca
     9181 gcttcaagga tcgctcgcgg ctcttaccag cctaacttcg atcattggac cgctgatcgt
     9241 cacggcgatt tatgccgcct cggcgagcac atggaacggg ttggcatgga ttgtaggcgc
     9301 cgccctatac cttgtctgcc tccccgcgtt gcgtcgcggt gcatggagcc gggccacctc
     9361 gacctgaatg gaagccggcg gcacctcgct aacggattca ccactccaag aattggagcc
     9421 aatcaattct tgcggagaac tgtgaatgcg caaaccaacc cttggcagaa catatccatc
     9481 gcgtccgcca tctccagcag ccgcacgcgg cgcatctcgg gcagcgttgg gtcctggcca
     9541 cgggtgcgca tgatcgtgct cctgtcgttg aggacccggc taggctggcg gggttgcctt
     9601 actggttagc agaatgaatc accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa
     9661 acgtctgcga cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg
     9721 gaaacgcgga agtcccctac gtgctgctga agttgcccgc aacagagagt ggaaccaacc
     9781 ggtgatacca cgatactatg actgagagtc aacgccatga gcggcctcat ttcttattct
     9841 gagttacaac agtccgcacc gctgtccggt agctccttcc ggtgggcgcg gggcatgact
     9901 atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca
     9961 gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgccc
    10021 tgcaccatta tgttccggat ctgcatcgca ggatgctgct ggctaccctg tggaacacct
    10081 acatctgtat taacgaagcg ctaaccgttt ttatcaggct ctgggaggca gaataaatga
    10141 tcatatcgtc aattattacc tccacgggga gagcctgagc aaactggcct caggcatttg
    10201 agaagcacac ggtcacactg cttccggtag tcaataaacc ggtaaaccag caatagacat
    10261 aagcggctat ttaacgaccc tgccctgaac cgacgaccgg gtcgaatttg ctttcgaatt
    10321 tctgccattc atccgcttat tatcacttat tcaggcgtag caccaggcgt ttaagggcac
    10381 caataactgc cttaaaaaaa ttacgccccg ccctgccact catcgcagta ctgttgtaat
    10441 tcattaagca ttctgccgac atggaagcca tcacagacgg catgatgaac ctgaatcgcc
    10501 agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatggtgaa aacgggggcg
    10561 aagaagttgt ccatattggc cacgtttaaa tcaaaactgg tgaaactcac ccagggattg
    10621 gctgagacga aaaacatatt ctcaataaac cctttaggga aataggccag gttttcaccg
    10681 taacacgcca catcttgcga atatatgtgt agaaactgcc ggaaatcgtc gtggtattca
    10741 ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca
    10801 ctatcccata tcaccagctc accgtctttc attgccatac g
//
