LOCUS       sequence-248980-       11695 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ptet-controlled expression of S. pyogenes Cas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11695)
  AUTHORS   Tapscott T, Guarnieri MT, Henard CA
  TITLE     Development of a CRISPR/Cas9 System for Methylococcus capsulatus In 
            Vivo Gene Editing.
  JOURNAL   Appl Environ Microbiol. 2019 May 16;85(11). pii: AEM.00340-19. doi: 
            10.1128/AEM.00340-19. Print 2019 Jun 1.
  PUBMED    30926729
REFERENCE   2  (bases 1 to 11695)
  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: "Appl 
            Environ Microbiol."; date: "2019-05-16"; volume: "85(11). pii: 
            AEM.00340-19. doi"; pages: " 10.1128/AEM.00340-19. Print 2019 Jun 1"
COMMENT     Sequence Label: pCAH01Sp::Cas9
FEATURES             Location/Qualifiers
     source          1..11695
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..24,25..29,30..35)
                     /label=tetA promoter
                     /note="contains a tet operator sequence that enables 
                     repression by TetR"
     protein_bind    6..24
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     CDS             64..4170
                     /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             64..4167
                     /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"
     rep_origin      4281..4736
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4368..4387)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4578..4599
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4763..4867
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4868..4936,4937..5728)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5086..5105)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             5735..6361
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(5811..5830)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             complement(6624..7415)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     CDS             complement(7870..9018)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     CDS             complement(9290..9661)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            9694..9803
                     /note="incP origin of transfer"
     rep_origin      9956..10587
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      10896..11484
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11385..11404
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttgacactct atcattgata gagttatttt accactccct atcagtggat agagaaaagt
       61 gaaatggata agaaatactc aataggctta gatatcggca caaatagcgt cggatgggcg
      121 gtgatcactg atgaatataa ggttccgtct aaaaagttca aggttctggg aaatacagac
      181 cgccacagta tcaaaaaaaa tcttataggg gctcttttat ttgacagtgg agagacagcg
      241 gaagcgactc gtctcaaacg gacagctcgt agaaggtata cacgtcggaa gaatcgtatt
      301 tgttatctac aggagatttt ttcaaatgag atggcgaaag tagatgatag tttctttcat
      361 cgacttgaag agtctttttt ggtggaagaa gacaagaagc atgaacgtca tcctattttt
      421 ggaaatatag tagatgaagt tgcttatcat gagaaatatc caactatcta tcatctgcga
      481 aaaaaattgg tagattctac tgataaagcg gatttgcgct taatctattt ggccttagcg
      541 catatgatta agtttcgtgg tcattttttg attgagggag atttaaatcc tgataatagt
      601 gatgtggaca aactatttat ccagttggta caaacctaca atcaattatt tgaagaaaac
      661 cctattaacg caagtggagt agatgctaaa gcgattcttt ctgcacgatt gagtaaatca
      721 agacgattag aaaatctcat tgctcagctc cccggtgaga agaaaaatgg cttatttggg
      781 aatctcattg ctttgtcatt gggtttgacc cctaatttta aatcaaattt tgatttggca
      841 gaagatgcta aattacagct ttcaaaagat acttacgatg atgatttaga taatttattg
      901 gcgcaaattg gagatcaata tgctgatttg tttttggcag ctaagaattt atcagatgct
      961 attttacttt cagatatcct aagagtaaat actgaaataa ctaaggctcc cctatcagct
     1021 tcaatgatta aacgctacga tgaacatcat caagacttga ctcttttaaa agctttagtt
     1081 cgacaacaac ttccagaaaa gtataaagaa atcttttttg atcaatcaaa aaacggatat
     1141 gcaggttata ttgatggggg agctagccaa gaagaatttt ataaatttat caaaccaatt
     1201 ttagaaaaaa tggatggtac tgaggaatta ttggtgaaac taaatcgtga agatttgctg
     1261 cgcaagcaac ggacctttga caacggctct attccccatc aaattcactt gggtgagctg
     1321 catgctattt tgagaagaca agaagacttt tatccatttt taaaagacaa tcgtgagaag
     1381 attgaaaaaa tcttgacttt tcgaattcct tattatgttg gtccattggc gcgtggcaat
     1441 agtcgttttg catggatgac tcggaagtct gaagaaacaa ttaccccatg gaattttgaa
     1501 gaagttgtcg ataaaggtgc ttcagctcaa tcatttattg aacgcatgac aaactttgat
     1561 aaaaatcttc caaatgaaaa agtactacca aaacatagtt tgctttatga gtattttacg
     1621 gtttataacg aattgacaaa ggtcaaatat gttactgaag gaatgcgaaa accagcattt
     1681 ctttcaggtg aacagaagaa agccattgtt gatttactct tcaaaacaaa tcgaaaagta
     1741 accgttaagc aattaaaaga agattatttc aaaaaaatag aatgttttga tagtgttgaa
     1801 atttcaggag ttgaagatag atttaatgct tcattaggta cctaccatga tttgctaaaa
     1861 attattaaag ataaagattt tttggataat gaagaaaatg aagatatctt agaggatatt
     1921 gttttaacat tgaccttatt tgaagatagg gagatgattg aggaaagact taaaacatat
     1981 gctcacctct ttgatgataa ggtgatgaaa cagcttaaac gtcgccgtta tactggttgg
     2041 ggacgtttgt ctcgaaaatt gattaatggt attagggata agcaatctgg caaaacaata
     2101 ttagattttt tgaaatcaga tggttttgcc aatcgcaatt ttatgcagct gatccatgat
     2161 gatagtttga catttaaaga agacattcaa aaagcacaag tgtctggaca aggcgatagt
     2221 ttacatgaac atattgcaaa tttagctggt agccctgcta ttaaaaaagg tattttacag
     2281 actgtaaaag ttgttgatga attggtcaaa gtaatggggc ggcataagcc agaaaatatc
     2341 gttattgaaa tggcacgtga aaatcagaca actcaaaagg gccagaaaaa ttcgcgagag
     2401 cgtatgaaac gaatcgaaga aggtatcaaa gaattaggaa gtcagattct taaagagcat
     2461 cctgttgaaa atactcaatt gcaaaatgaa aagctctatc tctattatct ccaaaatgga
     2521 agagacatgt atgtggacca agaattagat attaatcgtt taagtgatta tgatgtcgat
     2581 cacattgttc cacaaagttt ccttaaagac gattcaatag acaataaggt cttaacgcgt
     2641 tctgataaaa atcgtggtaa atcggataac gttccaagtg aagaagtagt caaaaagatg
     2701 aaaaactatt ggagacaact tctaaacgcc aagttaatca ctcaacgtaa gtttgataat
     2761 ttaacgaaag ctgaacgtgg aggtttgagt gaacttgata aagctggttt tatcaaacgc
     2821 caattggttg aaactcgcca aatcactaag catgtggcac aaattttgga tagtcgcatg
     2881 aatactaaat acgatgaaaa tgataaactt attcgagagg ttaaagtgat taccttaaaa
     2941 tctaaattag tttctgactt ccgaaaagat ttccaattct ataaagtacg tgagattaac
     3001 aattaccatc atgcccatga tgcgtatcta aatgccgtcg ttggaactgc tttgattaag
     3061 aaatatccaa aacttgaatc ggagtttgtc tatggtgatt ataaagttta tgatgttcgt
     3121 aaaatgattg ctaagtctga gcaagaaata ggcaaagcaa ccgcaaaata tttcttttac
     3181 tctaatatca tgaacttctt caaaacagaa attacacttg caaatggaga gattcgcaaa
     3241 cgccctctaa tcgaaactaa tggggaaact ggagaaattg tctgggataa agggcgagat
     3301 tttgccacag tgcgcaaagt attgtccatg ccccaagtca atattgtcaa gaaaacagaa
     3361 gtacagacag gcggattctc caaggagtca attttaccaa aaagaaattc ggacaagctt
     3421 attgctcgta aaaaagactg ggatccaaaa aaatatggtg gttttgatag tccaacggta
     3481 gcttattcag tcctagtggt tgctaaggtg gaaaaaggga aatcgaagaa gttaaaatcc
     3541 gttaaagagt tactagggat cacaattatg gaaagaagtt cctttgaaaa aaatccgatt
     3601 gactttttag aagctaaagg atataaggaa gttaaaaaag acttaatcat taaactacct
     3661 aaatatagtc tttttgagtt agaaaacggt cgtaaacgga tgctggctag tgccggagaa
     3721 ttacaaaaag gaaatgagct ggctctgcca agcaaatatg tgaatttttt atatttagct
     3781 agtcattatg aaaagttgaa gggtagtcca gaagataacg aacaaaaaca attgtttgtg
     3841 gagcagcata agcattattt agatgagatt attgagcaaa tcagtgaatt ttctaagcgt
     3901 gttattttag cagatgccaa tttagataaa gttcttagtg catataacaa acatagagac
     3961 aaaccaatac gtgaacaagc agaaaatatt attcatttat ttacgttgac gaatcttgga
     4021 gctcccgctg cttttaaata ttttgataca acaattgatc gtaaacgata tacgtctaca
     4081 aaagaagttt tagatgccac tcttatccat caatccatca ctggtcttta tgaaacacgc
     4141 attgatttga gtcagctagg aggtgactga gggcccggat ccggtaccga attcaagctt
     4201 gacctgtgaa gtgaaaaatg gcgcacattg tgcgacattt tttttgtctg ccgtttaccg
     4261 ctactgcgtc acggatctcc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4321 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     4381 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4441 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     4501 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4561 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     4621 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     4681 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttcagg
     4741 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     4801 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     4861 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     4921 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     4981 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     5041 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     5101 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     5161 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     5221 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     5281 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     5341 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     5401 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     5461 tctagcttcc cggcaacaat tgatagactg gatggaggcg gataaagttg caggaccact
     5521 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     5581 tggctctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     5641 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     5701 aggtgcctca ctgattaagc attggtagga attaatgatg tctcgtttag ataaaagtaa
     5761 agtgattaac agcgcattag agctgcttaa tgaggtcgga atcgaaggtt taacaacccg
     5821 taaactcgcc cagaagctag gtgtagagca gcctacattg tattggcatg taaaaaataa
     5881 gcgggctttg ctcgacgcct tagccattga gatgttagat aggcaccata ctcacttttg
     5941 ccctttagaa ggggaaagct ggcaagattt tttacgtaat aacgctaaaa gttttagatg
     6001 tgctttacta agtcatcgcg atggagcaaa agtacattta ggtacacggc ctacagaaaa
     6061 acagtatgaa actctcgaaa atcaattagc ctttttatgc caacaaggtt tttcactaga
     6121 gaatgcatta tatgcactca gcgcagtggg gcattttact ttaggttgcg tattggaaga
     6181 tcaagagcat caagtcgcta aagaagaaag ggaaacacct actactgata gtatgccgcc
     6241 attattacga caagctatcg aattatttga tcaccaaggt gcagagccag ccttcttatt
     6301 cggccttgaa ttgatcatat gcggattaga aaaacaactt aaatgtgaaa gtgggtctta
     6361 aaagcagcat aacctttttc cgtgatggta acttcactag tttaaaagga tctaggtgaa
     6421 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     6481 gtcagacccc gtagaaaatt gtcgggaaga tgcgtgatct gatccttcaa ctcagcaaaa
     6541 gttcgattta ttcaacaaag ccgccgtccc gtcaagtcag cgtaatgctc tgccagtgtt
     6601 acaaccaatt aaccaattct gatttatttg ccgactacct tggtgatctc gcctttcacg
     6661 tagtggacaa attcttccaa ctgatctgcg cgcgaggcca agcgatcttc ttcttgtcca
     6721 agataagcct gtctagcttc aagtatgacg ggctgatact gggccggcag gcgctccatt
     6781 gcccagtcgg cagcgacatc cttcggcgcg attttgccgg ttactgcgct gtaccaaatg
     6841 cgggacaacg taagcactac atttcgctca tcgccagccc agtcgggcgg cgagttccat
     6901 agcgttaagg tttcatttag cgcctcaaat agatcctgtt caggaaccgg atcaaagagt
     6961 tcctccgccg ctggacctac caaggcaacg ctatgttctc ttgcttttgt cagcaagata
     7021 gccagatcaa tgtcgatcgt ggctggctcg aagatacctg caagaatgtc attgcgctgc
     7081 cattctccaa attgcagttc gcgcttagct ggataacgcc acggaatgat gtcgtcgtgc
     7141 acaacaatgg tgacttctac agcgcggaga atctcgctct ctccagggga agccgaagtt
     7201 tccaaaaggt cgttgatcaa agctcgccgc gttgtttcat caagccttac ggtcaccgta
     7261 accagcaaat caatatcact gtgtggcttc aggccgccat ccactgcgga gccgtacaaa
     7321 tgtacggcca gcaacgtcgg ttcgagatgg cgctcgatga cgccaactac ctctgatagt
     7381 tgagtcgata cttcggcgat caccgcttcc ctcatgatgt ttaactttgt tttagggcga
     7441 ctgccctgct gcgtaacatc gttgctgctc cataacatca aacatcgacc cacggcgtaa
     7501 cgcgcttgct gcttggatgc ccgaggcata gactgtaccc caaaaaaaca gtcataacaa
     7561 gccattaaca ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgatga
     7621 tatattttta tcttgtgcaa tgtaacatca gagattttga gacacaacgt ggctttcgac
     7681 ttccggcaag ctatacgcgc cctagaattg tcaattttaa tcctctgttt atcggcagtt
     7741 cgtagagcgc gccgtgcgtc ccgagcgata ctgagcgaag caagtgcgtc gagcagtgcc
     7801 cgcttgttcc tgaaatgcca gtaaagcgct ggctgctgaa cccccagccg gaactgaccc
     7861 cacaaggccc tagcgtttgc aatgcaccag gtcatcattg acccaggcgt gttccaccag
     7921 gccgctgcct cgcaactctt cgcaggcttc gccgacctgc tcgcgccact tcttcacgcg
     7981 ggtggaatcc gatccgcaca tgaggcggaa ggtttccagc ttgagcgggt acggctcccg
     8041 gtgcgagctg aaatagtcga acatccgtcg ggccgtcggc gacagcttgc ggtacttctc
     8101 ccatatgaat ttcgtgtagt ggtcgccagc aaacagcacg acgatttcct cgtcgatcag
     8161 gacctggcaa cgggacgttt tcttgccacg gtccaggacg cggaagcggt gcagcagcga
     8221 caccgattcc aggtgcccaa cgcggtcgga cgtgaagccc atcgccgtcg cctgtaggcg
     8281 cgacaggcat tcctcggcct tcgtgtaata ccggccattg atcgaccagc ccaggtcctg
     8341 gcaaagctcg tagaacgtga aggtgatcgg ctcgccgata ggggtgcgct tcgcgtactc
     8401 caacacctgc tgccacacca gttcgtcatc gtcggcccgc agctcgacgc cggtgtaggt
     8461 gatcttcacg tccttgttga cgtggaaaat gaccttgttt tgcagcgcct cgcgcgggat
     8521 tttcttgttg cgcgtggtga acagggcaga gcgggccgtg tcgtttggca tcgctcgcat
     8581 cgtgtccggc cacggcgcaa tatcgaacaa ggaaagctgc atttccttga tctgctgctt
     8641 cgtgtgtttc agcaacgcgg cctgcttggc ctcgctgacc tgttttgcca ggtcctcgcc
     8701 ggcggttttt cgcttcttgg tcgtcatagt tcctcgcgtg tcgatggtca tcgacttcgc
     8761 caaacctgcc gcctcctgtt cgagacgacg cgaacgctcc acggcggccg atggcgcggg
     8821 cagggcaggg ggagccagtt gcacgctgtc gcgctcgatc ttggccgtag cttgctggac
     8881 catcgagccg acggactgga aggtttcgcg gggcgcacgc atgacggtgc ggcttgcgat
     8941 ggtttcggca tcctcggcgg aaaaccccgc gtcgatcagt tcttgcctgt atgccttccg
     9001 gtcaaacgtc cgattcattc accctccttg cgggattgcc ccgactcacg ccggggcaat
     9061 gtgcccttat tcctgatttg acccgcctgg tgccttggtg tccagataat ccaccttatc
     9121 ggcaatgaag tcggtcccgt agaccgtctg gccgtccttc tcgtacttgg tattccgaat
     9181 cttgccctgc acgaatacca gctccgcgaa gtcgctcttc ttgatggagc gcatggggac
     9241 gtgcttggca atcacgcgca ccccccggcc gttttagcgg ctaaaaaagt catggctctg
     9301 ccctcgggcg gaccacgccc atcatgacct tgccaagctc gtcctgcttc tcttcgatct
     9361 tcgccagcag ggcgaggatc gtggcatcac cgaaccgcgc cgtgcgcggg tcgtcggtga
     9421 gccagagttt cagcaggccg cccaggcggc ccaggtcgcc attgatgcgg gccagctcgc
     9481 ggacgtgctc atagtccacg acgcccgtga ttttgtagcc ctggccgacg gccagcaggt
     9541 aggcctacag gctcatgccg gccgccgccg ccttttcctc aatcgctctt cgttcgtctg
     9601 gaaggcagta caccttgata ggtgggctgc ccttcctggt tggcttggtt tcatcagcca
     9661 tccgcttgcc ctcatctgtt acgccggcgg tagccggcca gcctcgcaga gcaggattcc
     9721 cgttgagcac cgccaggtgc gaataaggga cagtgaagaa ggaacacccg ctcgcgggtg
     9781 ggcctacttc acctatcctg cccggctgac gccgttggat acaccaagga aagtctacac
     9841 gaaccctttg gcaaaatcct gtatatcgtg cgaaaaagga tggatatacc gaaaaaatcg
     9901 ctataatgac cccgaagcag ggttatgcag cggaaaagat ccgtcgaccc tttccgacgc
     9961 tcaccgggct ggttgccctc gccgctgggc tggcggccgt ctatggccct gcaaacgcgc
    10021 cagaaacgcc gtcgaagccg tgtgcgagac accgcggccg ccggcgttgt ggatacctcg
    10081 cggaaaactt ggccctcact gacagatgag gggcggacgt tgacacttga ggggccgact
    10141 cacccggcgc ggcgttgaca gatgaggggc aggctcgatt tcggccggcg acgtggagct
    10201 ggccagcctc gcaaatcggc gaaaacgcct gattttacgc gagtttccca cagatgatgt
    10261 ggacaagcct ggggataagt gccctgcggt attgacactt gaggggcgcg actactgaca
    10321 gatgaggggc gcgatccttg acacttgagg ggcagagtgc tgacagatga ggggcgcacc
    10381 tattgacatt tgaggggctg tccacaggca gaaaatccag catttgcaag ggtttccgcc
    10441 cgtttttcgg ccaccgctaa cctgtctttt aacctgcttt taaaccaata tttataaacc
    10501 ttgtttttaa ccagggctgc gccctgtgcg cgtgaccgcg cacgccgaag gggggtgccc
    10561 ccccttctcg aaccctcccg gcccgctaac gcgggcctcc catcccccca ggggctgcgc
    10621 ccctcggccg cgaacggcct caccccaaaa atggcagcca agctgacccg ctagggacgt
    10681 gaagtcgatt ccttcgatgg ttagcaatca aagactcaga gtgctgtctg gagcgtgaat
    10741 ctaacggtac gtatctcgat tgctcggtcg ctattcgcac tctgcgaaag ttcgtaccgc
    10801 tcattcacta ggttgcgaat catgaccaaa atcccttaac gtgagttttc gttccactga
    10861 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
    10921 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
    10981 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
    11041 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
    11101 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
    11161 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
    11221 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
    11281 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
    11341 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
    11401 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
    11461 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
    11521 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
    11581 cgtattaccg cctttgagtg agctgggcct ttttacggtt cctggccttt tgctggcctt
    11641 ttgctcacat gacccgacac catcgaatgg ccagatgatt aattcctaat ttttg
//
