LOCUS       sequence-371208-       13746 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  carrying CRISPR/Cas9 system for genome editing in Acidithiobacillus 
            ferridurans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13746)
  AUTHORS   Chen J, Liu Y, Mahadevan R
  TITLE     Genetic Engineering of Acidithiobacillus ferridurans Using CRISPR 
            Systems To Mitigate Toxic Release in Biomining.
  JOURNAL   Environ Sci Technol. 2023 Aug 22;57(33):12315-12324. doi: 
            10.1021/acs.est.3c02492. Epub 2023 Aug 9.
  PUBMED    37556825
REFERENCE   2  (bases 1 to 13746)
  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/acs.est.3c02492"; journalName: "Environ Sci Technol"; date:
            "2023-08-22- 22"; volume: "57"; issue: "33"; pages: "12315-12324"
COMMENT     Sequence Label: pAF-HdrB3
FEATURES             Location/Qualifiers
     source          1..13746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(149..154,155..170,171..177)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     CDS             198..4304
                     /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             198..4301
                     /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"
     terminator      4328..4399
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        4415..4449
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     misc_RNA        4470..4545
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      5700..5727
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5970..6821)
                     /codon_start=1
                     /gene="repC"
                     /product="replication protein C of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepC
                     /translation="MVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
                     GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
                     TADAMVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
                     YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
                     TGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA
                     AG"
     CDS             complement(6808..7647)
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein A of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepA
                     /translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
                     LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
                     GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
                     EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
                     WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
     CDS             complement(8158..9129)
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein B of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepB
                     /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
                     QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
                     EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
     oriT            10368..10455
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     rep_origin      10797..11191
                     /label=RSF1010 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     RSF1010; requires the RSF1010 RepA/B/C proteins for 
                     replication (Scholz et al., 1989)"
     promoter        complement(join(13686..13691,13692..13708,13709..13714))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 aatatcgtcg gagataaata taataattcc acggactata gactatacta gtatactccg
       61 tctactgtac gatacacttc cgctcaggtc cttgtccttt aacgaggatt gttacgtacg
      121 ctaatggcgt caaaacaaag actctagatt gacaattaat catcggctcg tataatgttt
      181 caccttatca tcaagatatg gataagaaat actcaatagg cttagatatc ggcacaaata
      241 gcgtcggatg ggcggtgatc actgatgaat ataaggttcc gtctaaaaag ttcaaggttc
      301 tgggaaatac agaccgccac agtatcaaaa aaaatcttat aggggctctt ttatttgaca
      361 gtggagagac agcggaagcg actcgtctca aacggacagc tcgtagaagg tatacacgtc
      421 ggaagaatcg tatttgttat ctacaggaga ttttttcaaa tgagatggcg aaagtagatg
      481 atagtttctt tcatcgactt gaagagtctt ttttggtgga agaagacaag aagcatgaac
      541 gtcatcctat ttttggaaat atagtagatg aagttgctta tcatgagaaa tatccaacta
      601 tctatcatct gcgaaaaaaa ttggtagatt ctactgataa agcggatttg cgcttaatct
      661 atttggcctt agcgcatatg attaagtttc gtggtcattt tttgattgag ggagatttaa
      721 atcctgataa tagtgatgtg gacaaactat ttatccagtt ggtacaaacc tacaatcaat
      781 tatttgaaga aaaccctatt aacgcaagtg gagtagatgc taaagcgatt ctttctgcac
      841 gattgagtaa atcaagacga ttagaaaatc tcattgctca gctccccggt gagaagaaaa
      901 atggcttatt tgggaatctc attgctttgt cattgggttt gacccctaat tttaaatcaa
      961 attttgattt ggcagaagat gctaaattac agctttcaaa agatacttac gatgatgatt
     1021 tagataattt attggcgcaa attggagatc aatatgctga tttgtttttg gcagctaaga
     1081 atttatcaga tgctatttta ctttcagata tcctaagagt aaatactgaa ataactaagg
     1141 ctcccctatc agcttcaatg attaaacgct acgatgaaca tcatcaagac ttgactcttt
     1201 taaaagcttt agttcgacaa caacttccag aaaagtataa agaaatcttt tttgatcaat
     1261 caaaaaacgg atatgcaggt tatattgatg ggggagctag ccaagaagaa ttttataaat
     1321 ttatcaaacc aattttagaa aaaatggatg gtactgagga attattggtg aaactaaatc
     1381 gtgaagattt gctgcgcaag caacggacct ttgacaacgg ctctattccc catcaaattc
     1441 acttgggtga gctgcatgct attttgagaa gacaagaaga cttttatcca tttttaaaag
     1501 acaatcgtga gaagattgaa aaaatcttga cttttcgaat tccttattat gttggtccat
     1561 tggcgcgtgg caatagtcgt tttgcatgga tgactcggaa gtctgaagaa acaattaccc
     1621 catggaattt tgaagaagtt gtcgataaag gtgcttcagc tcaatcattt attgaacgca
     1681 tgacaaactt tgataaaaat cttccaaatg aaaaagtact accaaaacat agtttgcttt
     1741 atgagtattt tacggtttat aacgaattga caaaggtcaa atatgttact gaaggaatgc
     1801 gaaaaccagc atttctttca ggtgaacaga agaaagccat tgttgattta ctcttcaaaa
     1861 caaatcgaaa agtaaccgtt aagcaattaa aagaagatta tttcaaaaaa atagaatgtt
     1921 ttgatagtgt tgaaatttca ggagttgaag atagatttaa tgcttcatta ggtacctacc
     1981 atgatttgct aaaaattatt aaagataaag attttttgga taatgaagaa aatgaagata
     2041 tcttagagga tattgtttta acattgacct tatttgaaga tagggagatg attgaggaaa
     2101 gacttaaaac atatgctcac ctctttgatg ataaggtgat gaaacagctt aaacgtcgcc
     2161 gttatactgg ttggggacgt ttgtctcgaa aattgattaa tggtattagg gataagcaat
     2221 ctggcaaaac aatattagat tttttgaaat cagatggttt tgccaatcgc aattttatgc
     2281 agctgatcca tgatgatagt ttgacattta aagaagacat tcaaaaagca caagtgtctg
     2341 gacaaggcga tagtttacat gaacatattg caaatttagc tggtagccct gctattaaaa
     2401 aaggtatttt acagactgta aaagttgttg atgaattggt caaagtaatg gggcggcata
     2461 agccagaaaa tatcgttatt gaaatggcac gtgaaaatca gacaactcaa aagggccaga
     2521 aaaattcgcg agagcgtatg aaacgaatcg aagaaggtat caaagaatta ggaagtcaga
     2581 ttcttaaaga gcatcctgtt gaaaatactc aattgcaaaa tgaaaagctc tatctctatt
     2641 atctccaaaa tggaagagac atgtatgtgg accaagaatt agatattaat cgtttaagtg
     2701 attatgatgt cgatcacatt gttccacaaa gtttccttaa agacgattca atagacaata
     2761 aggtcttaac gcgttctgat aaaaatcgtg gtaaatcgga taacgttcca agtgaagaag
     2821 tagtcaaaaa gatgaaaaac tattggagac aacttctaaa cgccaagtta atcactcaac
     2881 gtaagtttga taatttaacg aaagctgaac gtggaggttt gagtgaactt gataaagctg
     2941 gttttatcaa acgccaattg gttgaaactc gccaaatcac taagcatgtg gcacaaattt
     3001 tggatagtcg catgaatact aaatacgatg aaaatgataa acttattcga gaggttaaag
     3061 tgattacctt aaaatctaaa ttagtttctg acttccgaaa agatttccaa ttctataaag
     3121 tacgtgagat taacaattac catcatgccc atgatgcgta tctaaatgcc gtcgttggaa
     3181 ctgctttgat taagaaatat ccaaaacttg aatcggagtt tgtctatggt gattataaag
     3241 tttatgatgt tcgtaaaatg attgctaagt ctgagcaaga aataggcaaa gcaaccgcaa
     3301 aatatttctt ttactctaat atcatgaact tcttcaaaac agaaattaca cttgcaaatg
     3361 gagagattcg caaacgccct ctaatcgaaa ctaatgggga aactggagaa attgtctggg
     3421 ataaagggcg agattttgcc acagtgcgca aagtattgtc catgccccaa gtcaatattg
     3481 tcaagaaaac agaagtacag acaggcggat tctccaagga gtcaatttta ccaaaaagaa
     3541 attcggacaa gcttattgct cgtaaaaaag actgggatcc aaaaaaatat ggtggttttg
     3601 atagtccaac ggtagcttat tcagtcctag tggttgctaa ggtggaaaaa gggaaatcga
     3661 agaagttaaa atccgttaaa gagttactag ggatcacaat tatggaaaga agttcctttg
     3721 aaaaaaatcc gattgacttt ttagaagcta aaggatataa ggaagttaaa aaagacttaa
     3781 tcattaaact acctaaatat agtctttttg agttagaaaa cggtcgtaaa cggatgctgg
     3841 ctagtgccgg agaattacaa aaaggaaatg agctggctct gccaagcaaa tatgtgaatt
     3901 ttttatattt agctagtcat tatgaaaagt tgaagggtag tccagaagat aacgaacaaa
     3961 aacaattgtt tgtggagcag cataagcatt atttagatga gattattgag caaatcagtg
     4021 aattttctaa gcgtgttatt ttagcagatg ccaatttaga taaagttctt agtgcatata
     4081 acaaacatag agacaaacca atacgtgaac aagcagaaaa tattattcat ttatttacgt
     4141 tgacgaatct tggagctccc gctgctttta aatattttga tacaacaatt gatcgtaaac
     4201 gatatacgtc tacaaaagaa gttttagatg ccactcttat ccatcaatcc atcactggtc
     4261 tttatgaaac acgcattgat ttgagtcagc taggaggtga ctaactcgag taaggatctc
     4321 caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     4381 tttgtcggtg aacgctctct actcccgggg atccttgaca gctagctcag tcctaggtat
     4441 aatactagtg acaacaacag cacgcaacag ttttagagct agaaatagca agttaaaata
     4501 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt ttgaattctg
     4561 taggctagtc cgggcgcgcc gaatgtagca tcagcaaaca ctgttcaggc tgaagcagga
     4621 ccggatcggc gcccgcagcg acaaccgcat ccagcaagcg gcgggtcgaa tagagctcca
     4681 catggcgcga aagaatggct acgcggggca ccgcaaccat gcctaggcta catcctcatg
     4741 gaaactgagc aggggcaact cgccgtgccc gatggatcgc tcgatgaaac ctgcccgccc
     4801 cggcgaccgc tgctccagcg ctgtccttgc cagggcgggc gcagcgctga tcgcggcaaa
     4861 gagtacctcc agttcccagg gatcgacccg cagcgtgcca aagggattat cccgcgttgc
     4921 cggacgccag aagagattcg gcaaccatcc tttgttgcgc actgcggggg ccagataaaa
     4981 cgcgcgacaa cgtgcggcta cctccgccga cgcgggaatc gaaatcacat cctggttttc
     5041 cataataccc ctttcaagtc gttgctgatg cgggccgcca ccccaatcga caaaagcctc
     5101 ccctccggac atgtttttcg ggtatgctta ggcacgcccg ccacgacagg cgatggcttt
     5161 gcccgtttaa acaggaggat gacagtggac ccgaaggccc tcaatgcccg ttgtgtagaa
     5221 ctttttcaga gcccggatgt gcgcctgcgg atgtggaatg cgcggatgtt ctggcaggtt
     5281 ggcgatcaga tgaatgtcgc gccgaccgca ctcaccgatc ccaaggtgga tacctgcgaa
     5341 ctggaagtca tgctcagcgc cgccgcactg accgattccc agtgtgccgc tgaactggac
     5401 aaacgcgagc ccggccgcgc cgccttcatt cagcgccagg tccgggaagg catgcgaccg
     5461 cttctgcgtc ctgcgcaata gtcaaggtcg caggcattgc gtagtagttg caggcccccg
     5521 caacatgggt ggtcgccagc gtagcggccc cctgtgcggc gacgttgacc ggcaacgtac
     5581 agcgaattcc gcaggcattg cggtaccgta gttgcgggcc ccaatagcgg ccccccgtcg
     5641 acctgcaaaa gcttgcggcc ggtcgtgact gggaaaaccc tggcgactag cttggcactg
     5701 gctcaccttc gggtgggcct ttctgcgttt atacctagga agcttacttg ttaacttcag
     5761 gacctgcagg agcagaagag catacatctg gaagcaaagc caggaaagcg gcctatggag
     5821 ctgtgcggca gcgctcagta ggcaattttt caaaatattg ttaagccttt tctgagcatg
     5881 gtatttttca tggtattacc aattagcagg aaaataagcc attgaatata aaagataaaa
     5941 atgtcttgtt tacaatagag tggggggggt cagcctgccg ccttgggccg ggtgatgtcg
     6001 tacttgcccg ccgcgaactc ggttaccgtc cagcccagcg cgaccagctc cggcaacgcc
     6061 tcgcgcaccc gctggcggcg cttgcgcatg gtcgaaccac tggcctctga cggccagaca
     6121 tagccgcaca aggtatctat ggaagccttg ccggttttgc cggggtcgat ccagccacac
     6181 agccgctggt gcagcaggcg ggcggtttcg ctgtccagcg cccgcacctc gtccatgctg
     6241 atgcgcacat gctggccgcc acccatgacg gcctgcgcga tcaaggggtt cagggccacg
     6301 tacaggcgcc cgtccgcctc gtcgctggcg tactccgaca gcagccgaaa cccctgccgc
     6361 ttgcggccat tctgggcgat gatggatacc ttccaaaggc gctcgatgca gtcctgtatg
     6421 tgcttgagcg ccccaccact atcgacctct gccccgattt cctttgccag cgcccgatag
     6481 ctacctttga ccaccatggc atcagcggtg acggcctccc acttgggttc caggaacagc
     6541 cggagctgcc gtccgccttc ggtcttgggt tccgggccaa gcactaggcc attaggccca
     6601 gccatggcca ccagcccttg caggatgcgc agatcatcag cgcccagcgg ctccgggccg
     6661 ctgaactcga tccgcttgcc gtcgccgtag tcatacgtca cgtccagctt gctgcgcttg
     6721 cgctcgcccc gcttgagggc acggaacagg ccgggggcca gacagtgcgc cgggtcgtgc
     6781 cggacgtggc tgaggctgtg cttgttctta ggcttcacca cggggcaccc ccttgctctt
     6841 gcgctgcctc tccagcacgg cgggcttgag caccccgccg tcatgccgcc tgaaccaccg
     6901 atcagcgaac ggtgcgccat agttggcctt gctcacaccg aagcggacga agaaccggcg
     6961 ctggtcgtcg tccacacccc attcctcggc ctcggcgctg gtcatgctcg acaggtagga
     7021 ctgccagcgg atgttatcga ccagtaccga gctgccccgg ctggcctgct gctggtcgcc
     7081 tgcgcccatc atggccgcgc ccttgctggc atggtgcagg aacacgatag agcacccggt
     7141 atcggcggcg atggcctcca tgcgaccgat gacctgggcc atggggccgc tggcgttttc
     7201 ttcctcgatg tggaaccggc gcagcgtgtc cagcaccatc aggcggcggc cctcggcggc
     7261 gcgcttgagg ccgtcgaacc actccggggc catgatgttg ggcaggctgc cgatcagcgg
     7321 ctggatcagc aggccgtcag ccacggcttg ccgttcctcg gcgctgaggt gcgccccaag
     7381 ggcgtgcagg cggtgatgaa tggcggtggg cgggtcttcg gcgggcaggt agatcaccgg
     7441 gccggtgggc agttcgccca cctccagcag atccggcccg cctgcaatct gtgcggccag
     7501 ttgcagggcc agcatggatt taccggcacc accgggcgac accagcgccc cgaccgtacc
     7561 ggccaccatg ttgggcaaaa cgtagtccag cggtggcggc gctgctgcga acgcctccag
     7621 aatattgata ggcttatggg tagccattga ttgcctcctt tgcaggcagt tggtggttag
     7681 gcgctggcgg ggtcactacc cccgccctgc gccgctctga gttcttccag gcactcgcgc
     7741 agcgcctcgt attcgtcgtc ggtcagccag aacttgcgct gacgcatccc tttggccttc
     7801 atgcgctcgg catatcgcgc ttggcgtaca gcgtcagggc tggccagcag gtcgccggtc
     7861 tgcttgtcct tttggtcttt catatcagtc accgagaaac ttgccggggc cgaaaggctt
     7921 gtcttcgcgg aacaaggaca aggtgcagcc gtcaaggtta aggctggcca tatcagcgac
     7981 tgaaaagcgg ccagcctcgg ccttgtttga cgtataacca aagccaccgg gcaaccaata
     8041 gcccttgtca cttttgatca ggtagaccga ccctgaagcg cttttttcgt attccataaa
     8101 acccccttct gtgcgtgagt actcatagta taacaggcgt gagtaccaac gcaagcacta
     8161 catgctgaaa tctggcccgc ccctgtccat gcctcgctgg cggggtgccg gtgcccgtgc
     8221 cagctcggcc cgcgcaagct ggacgctggg cagacccatg accttgctga cggtgcgctc
     8281 gatgtaatcc gcttcgtggc cgggcttgcg ctctgccagc gctgggctgg cctcggccat
     8341 ggccttgccg atttcctcgg cactgcggcc ccggctggcc agcttctgcg cggcgataaa
     8401 gtcgcacttg ctgaggtcat caccgaagcg cttgaccagc ccggccatct cgctgcggta
     8461 ctcgtccagc gccgtgcgcc ggtggcggct aagctgccgc tcgggcagtt cgaggctggc
     8521 cagcctgcgg gccttctcct gctgccgctg ggcctgctcg atctgctggc cagcctgctg
     8581 caccagcgcc gggccagcgg tggcggtctt gcccttggat tcacgcagca gcacccacgg
     8641 ctgataaccg gcgcgggtgg tgtgcttgtc cttgcggttg gtgaagcccg ccaagcggcc
     8701 atagtggcgg ctgtcggcgc tggccgggtc ggcgtcgtac tcgctggcca gcgtccgggc
     8761 aatctgcccc cgaagttcac cgcctgcggc gtcggccacc ttgacccatg cctgatagtt
     8821 cttcgggctg gtttccacta ccagggcagg ctcccggccc tcggctttca tgtcatccag
     8881 gtcaaactcg ctgaggtcgt ccaccagcac cagaccatgc cgctcctgct cggcgggcct
     8941 gatatacacg tcattgccct gggcattcat ccgcttgagc catggcgtgt tctggagcac
     9001 ttcggcggct gaccattccc ggttcatcat ctggccggtg gtggcgtccc tgacgccgat
     9061 atcgaagcgc tcacagccca tggccttgag ctgtcggcct atggcctgca aagtcctgtc
     9121 gttcttcatc gggccaccaa gcgcagccag atcgagccgt cctcggttgt cagtggcgtc
     9181 aggtcgagca agagcaacga tgcgatcagc agcaccaccg taggcatcat ggaagccagc
     9241 atcacggtta gccatagctt ccagtgccac ccccgcgacg cgctccgggc gctctgcgcg
     9301 gcgctgctca cctcggcggc tacctcccgc aactctttgg ccagctccac ccatgccgcc
     9361 cctgtctggc gctgggcttt cagccactcc gccgcctgcg cctcgctggc ctgctgggtc
     9421 tggctcatga cctgccgggc ttcgtcggcc agtgtcgcca tgctctgggc cagcggttcg
     9481 atctgctccg ctaactcgtt gatgcctctg gatttcttca ctctgtcgat tgcgttcatg
     9541 gtctattgcc tcccggtatt cctgtaagtc gatgatctgg gcgttggcgg tgtcgatgtt
     9601 cagggccacg tctgcccggt cggtgcggat gccccggcct tccatctcca ccacgttcgg
     9661 ccccaggtga acaccgggca ggcgctcgat gccctgcgcc tcaagtgttc tgtggtcaat
     9721 gcgggcgtcg tggccagccc gctctaatgc ccggttggca tggtcggccc atgcctcgcg
     9781 ggtctgctca agccatgcct tgggcttgag cgcttcggtc ttctgtgccc cgcccttctc
     9841 cggggtcttg ccgttgtacc gcttgaacca ctgagcggcg ggccgctcga tgccgtcatt
     9901 gatccgctcg gagatcatca ggtggcagtg cgggttctcg ccgccaccgg catggatggc
     9961 cagcgtatac ggcaggcgct cggcaccggt caggtgctgg gcgaactcgg acgccagcgc
    10021 cttctgctgg tcgagggtca gctcgaccgg cagggcaaat tcgacctcct tgaacagccg
    10081 cccattggcg cgttcataca ggtcggcagc atcccagtag tcggcgggcc gctcgacgaa
    10141 ctccggcatg tgcccggatt cggcgtgcaa gacttcatcc atgtcgcggg catacttgcc
    10201 ttcgcgctgg atgtagtcgg ccttggccct ggccgattgg ccgcccgacc tgctgccggt
    10261 tttcgccgta aggtgataaa tcgccatgct gcctcgctgt tgcttttgct tttcggctcc
    10321 atgcaatggc cctcggagag cgcaccgccc gaagggtggc cgttaggcca gtttctcgaa
    10381 gagaaaccgg taagtgcgcc ctcccctaca aagtagggtc gggattgccg ccgctgtgcc
    10441 tccatgatag cctacgagac agcacattaa caatggggtg tcaagatggt taaggggagc
    10501 aacaaggcgg cggatcggct ggccaagctc gaagaacaac gagcgcgaat caatgccgaa
    10561 attcagcggg tgcgggcaag ggaacagcag caagagcgca agaacgaaac aaggcgcaag
    10621 gtgctggtgg gggccatgat tttggccaag gtgaacagca gcgagtggcc ggaggatcgg
    10681 ctcatggcgg caatggatgc gtaccttgaa cgcgaccacg accgcgcctt gttcggtctg
    10741 ccgccacgcc agaaggatga gccgggctga atgatcgacc gagacaggcc ctgcggggct
    10801 gcacacgcgc ccccaccctt cgggtagggg gaaaggccgc taaagcggct aaaagcgctc
    10861 cagcgtattt ctgcggggtt tggtgtgggg tttagcgggc tttgcccgcc tttccccctg
    10921 ccgcgcagcg gtggggcggt gtgtagccta gcgcagcgaa tagaccagct atccggcctc
    10981 tggccgggca tattgggcaa gggcagcagc gccccacaag ggcgctgata accgcgccta
    11041 gtggattatt cttagataat catggatgga tttttccaac accccgccag cccccgcccc
    11101 tgctgggttt gcaggtttgg gggcgtgaca gttattgcag gggttcgtga cagttattgc
    11161 aggggggcgt gacagttatt gcaggggttc gtgacagtta gtacgggagt gacgggcact
    11221 ggctggcaat gtctagcaac ggcaggcatt tcggctgagg gtaaaagaac tttccgctaa
    11281 gcgatagact gtatgtaaac acagtattgc aaggacgcgg aacatgcctc atgtggcggc
    11341 caggacggcc agccgggatc gggatactgg tcgttaccag agccaccgac ccgagcaaac
    11401 ccttctctat cagatcgttg acgagtatta cccggcattc gctgcgctta tggcagagca
    11461 gggaaaggaa ttgccgggct atgtgcaacg ggaatttgaa gaatttctcc aatgcgggcg
    11521 gctggagcat ggctttctac gggttcgctg cgagtcttgc cacgccgagc acctggtcgc
    11581 tttcagctgt aagcgtcgcg gtttctgccc gagctgtggg gcgcggcgga tggccgaaag
    11641 tgccgccttg ctggttgatg aagtactgcc tgaacaaccc atgcgtcagt gggtgttgag
    11701 cttcccgttt cagctgcgtt tcctgtttgg ggtcgtttgc gggaaggggc ggaatcctac
    11761 gctaaggctt tggccagcga tattctccgg tgagattgat gtgttcccag gggataggag
    11821 aagtcgcttg atatctagta tgacgtctgt cgcacctgct tgatcgcggc cgcgatagct
    11881 agatcgcgtt gctcctcttc tccatccgcg ttccaagctg cggaaaggca cccataagcg
    11941 tacgcctggt cgagcaggcg acgcggatcg acgtccagcg cacgagagaa tgcgtccgcc
    12001 atctgtgcaa tgcgtctagg atcgagacaa aggtcgtctc tgtcagccgg atcgtagaac
    12061 atattggcgg cgccaaagcc cacttcaccg accagaccga cgggatctat caccagccag
    12121 ccgcgactgg agaacatgat gttttcatga tgcagatcgc catgtagccc acgcagttcc
    12181 gaggcattgc tcatcatttg atcggctata atcgccgcgt ggacgtagtc agtttgacaa
    12241 cctgcgtttt gatcatcgcg cgcccgctga aacaaagctg caaagcgatc ccggatcggg
    12301 agaagggcag aaggcagggg ttcctcagat gcggcataca gcttcgccat tagttccgct
    12361 gcaatttcgg tcgcctggta gtcgccgtgc tcggcaacga tgtgagagag cattcgctcc
    12421 ccggcatatt cgagcaacat cagattgttc tcacgaccga gcaaccggac tgctcccctc
    12481 ccattgcgcc ataccagata gtcggccccg cgcagttcat cagcaatgtc ttctataggt
    12541 ttcaatccct tgacgattgc aggagtcccg tctggcaatg aaactttcca aacgaggctg
    12601 gaaaaggtgt ccgcaatgag aacaggttgc gaaacgtgcc aatgagcagg aaaaacaggc
    12661 ggcatgaaca tcaaccccaa gtcagagggt ccaatcgcag atagaaggca aggcgttcgc
    12721 ggtcgggggc ttcgatcccc aatacattga ataggacagc gaaggcgcgc tctgcttcat
    12781 ctggcgctgc ccagttctct tcggcgttag caatcatgag tgccaaatcg gcatagcgat
    12841 ctgctgttcc gagccgccca aggtcgatca gacccgtgca ttgaagagtt ttagggtcca
    12901 ccatgaagtt cggcatgcag ggatcaccat ggcaaacaac catatcggtg cgctcttggt
    12961 cgagccgcac cggtagctct cgttcgacac gagccaaaag atcgagctgc ggcgtactct
    13021 tgtcctcgtc cggtaagaag tcgggattga cggcattgcg ggacaccaca tcaacggcgc
    13081 gtccgaacat tcgcgacagc ctgcgctcaa acggacattg atcaaccgat aggctgtgaa
    13141 cagcgccaag ttgctgcccc attgacggcc acgctttgag caaatccgct ccagacagat
    13201 cagccgccgg tactcccgga attgccgtta tcaccaagca tgcaccctcc tgttcctcct
    13261 gccagttgat gacctcgggg caagccacac ctcgaccttt gagccaaatg aggcggtcac
    13321 gctctccagc gagctcaccg cggcgggaag caggtgcgat tttcgcgaag gcatgcccgt
    13381 caccacgtcg aaaaacaaaa tcaccagatt ctccgcctct gacaggcaac cagtcagaat
    13441 gcgattcacc aaaaaaaata ttagttcgat tcaatggagg ttccttcagt tttctgatga
    13501 agcgcgaata tagagaaata tcccgaatgt gcagttatcc cgcaagaggc ccggcagtac
    13561 cggcataacc aagcctatgc ctacagcatc cagggtgacg gtgccgagga tgacgatgag
    13621 cgcattgtta gatttcatac acggtgcctg actgcgttag caatttaact gtgataaact
    13681 accgcattaa agcttatcga tgataagctg tcaaacatga gaattcccct ggatttcact
    13741 gatgag
//
