LOCUS       sequence-248978-       11695 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ptet-controlled expression of S. pyogenes Cas9D10A variant.
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::Cas9D10A
FEATURES             Location/Qualifiers
     source          1..11695
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(212..217,218..235,236..240,241..246)
                     /label=tetA promoter
                     /note="contains a tet operator sequence that enables 
                     repression by TetR"
     protein_bind    217..235
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     CDS             join(275..301,302..304,305..4381)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     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      4492..4947
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4579..4598)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4789..4810
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4974..5078
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5079..5147,5148..5939)
                     /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(5297..5316)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             5946..6572
                     /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(6022..6041)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             complement(6835..7626)
                     /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(8081..9229)
                     /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(9501..9872)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            9905..10014
                     /note="incP origin of transfer"
     rep_origin      10167..10798
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      11107..11695
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11596..11615
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
      121 gggccttttt acggttcctg gccttttgct ggccttttgc tcacatgacc cgacaccatc
      181 gaatggccag atgattaatt cctaattttt gttgacactc tatcattgat agagttattt
      241 taccactccc tatcagtgga tagagaaaag tgaaatggat aagaaatact caataggctt
      301 agccatcggc acaaatagcg tcggatgggc ggtgatcact gatgaatata aggttccgtc
      361 taaaaagttc aaggttctgg gaaatacaga ccgccacagt atcaaaaaaa atcttatagg
      421 ggctctttta tttgacagtg gagagacagc ggaagcgact cgtctcaaac ggacagctcg
      481 tagaaggtat acacgtcgga agaatcgtat ttgttatcta caggagattt tttcaaatga
      541 gatggcgaaa gtagatgata gtttctttca tcgacttgaa gagtcttttt tggtggaaga
      601 agacaagaag catgaacgtc atcctatttt tggaaatata gtagatgaag ttgcttatca
      661 tgagaaatat ccaactatct atcatctgcg aaaaaaattg gtagattcta ctgataaagc
      721 ggatttgcgc ttaatctatt tggccttagc gcatatgatt aagtttcgtg gtcatttttt
      781 gattgaggga gatttaaatc ctgataatag tgatgtggac aaactattta tccagttggt
      841 acaaacctac aatcaattat ttgaagaaaa ccctattaac gcaagtggag tagatgctaa
      901 agcgattctt tctgcacgat tgagtaaatc aagacgatta gaaaatctca ttgctcagct
      961 ccccggtgag aagaaaaatg gcttatttgg gaatctcatt gctttgtcat tgggtttgac
     1021 ccctaatttt aaatcaaatt ttgatttggc agaagatgct aaattacagc tttcaaaaga
     1081 tacttacgat gatgatttag ataatttatt ggcgcaaatt ggagatcaat atgctgattt
     1141 gtttttggca gctaagaatt tatcagatgc tattttactt tcagatatcc taagagtaaa
     1201 tactgaaata actaaggctc ccctatcagc ttcaatgatt aaacgctacg atgaacatca
     1261 tcaagacttg actcttttaa aagctttagt tcgacaacaa cttccagaaa agtataaaga
     1321 aatctttttt gatcaatcaa aaaacggata tgcaggttat attgatgggg gagctagcca
     1381 agaagaattt tataaattta tcaaaccaat tttagaaaaa atggatggta ctgaggaatt
     1441 attggtgaaa ctaaatcgtg aagatttgct gcgcaagcaa cggacctttg acaacggctc
     1501 tattccccat caaattcact tgggtgagct gcatgctatt ttgagaagac aagaagactt
     1561 ttatccattt ttaaaagaca atcgtgagaa gattgaaaaa atcttgactt ttcgaattcc
     1621 ttattatgtt ggtccattgg cgcgtggcaa tagtcgtttt gcatggatga ctcggaagtc
     1681 tgaagaaaca attaccccat ggaattttga agaagttgtc gataaaggtg cttcagctca
     1741 atcatttatt gaacgcatga caaactttga taaaaatctt ccaaatgaaa aagtactacc
     1801 aaaacatagt ttgctttatg agtattttac ggtttataac gaattgacaa aggtcaaata
     1861 tgttactgaa ggaatgcgaa aaccagcatt tctttcaggt gaacagaaga aagccattgt
     1921 tgatttactc ttcaaaacaa atcgaaaagt aaccgttaag caattaaaag aagattattt
     1981 caaaaaaata gaatgttttg atagtgttga aatttcagga gttgaagata gatttaatgc
     2041 ttcattaggt acctaccatg atttgctaaa aattattaaa gataaagatt ttttggataa
     2101 tgaagaaaat gaagatatct tagaggatat tgttttaaca ttgaccttat ttgaagatag
     2161 ggagatgatt gaggaaagac ttaaaacata tgctcacctc tttgatgata aggtgatgaa
     2221 acagcttaaa cgtcgccgtt atactggttg gggacgtttg tctcgaaaat tgattaatgg
     2281 tattagggat aagcaatctg gcaaaacaat attagatttt ttgaaatcag atggttttgc
     2341 caatcgcaat tttatgcagc tgatccatga tgatagtttg acatttaaag aagacattca
     2401 aaaagcacaa gtgtctggac aaggcgatag tttacatgaa catattgcaa atttagctgg
     2461 tagccctgct attaaaaaag gtattttaca gactgtaaaa gttgttgatg aattggtcaa
     2521 agtaatgggg cggcataagc cagaaaatat cgttattgaa atggcacgtg aaaatcagac
     2581 aactcaaaag ggccagaaaa attcgcgaga gcgtatgaaa cgaatcgaag aaggtatcaa
     2641 agaattagga agtcagattc ttaaagagca tcctgttgaa aatactcaat tgcaaaatga
     2701 aaagctctat ctctattatc tccaaaatgg aagagacatg tatgtggacc aagaattaga
     2761 tattaatcgt ttaagtgatt atgatgtcga tcacattgtt ccacaaagtt tccttaaaga
     2821 cgattcaata gacaataagg tcttaacgcg ttctgataaa aatcgtggta aatcggataa
     2881 cgttccaagt gaagaagtag tcaaaaagat gaaaaactat tggagacaac ttctaaacgc
     2941 caagttaatc actcaacgta agtttgataa tttaacgaaa gctgaacgtg gaggtttgag
     3001 tgaacttgat aaagctggtt ttatcaaacg ccaattggtt gaaactcgcc aaatcactaa
     3061 gcatgtggca caaattttgg atagtcgcat gaatactaaa tacgatgaaa atgataaact
     3121 tattcgagag gttaaagtga ttaccttaaa atctaaatta gtttctgact tccgaaaaga
     3181 tttccaattc tataaagtac gtgagattaa caattaccat catgcccatg atgcgtatct
     3241 aaatgccgtc gttggaactg ctttgattaa gaaatatcca aaacttgaat cggagtttgt
     3301 ctatggtgat tataaagttt atgatgttcg taaaatgatt gctaagtctg agcaagaaat
     3361 aggcaaagca accgcaaaat atttctttta ctctaatatc atgaacttct tcaaaacaga
     3421 aattacactt gcaaatggag agattcgcaa acgccctcta atcgaaacta atggggaaac
     3481 tggagaaatt gtctgggata aagggcgaga ttttgccaca gtgcgcaaag tattgtccat
     3541 gccccaagtc aatattgtca agaaaacaga agtacagaca ggcggattct ccaaggagtc
     3601 aattttacca aaaagaaatt cggacaagct tattgctcgt aaaaaagact gggatccaaa
     3661 aaaatatggt ggttttgata gtccaacggt agcttattca gtcctagtgg ttgctaaggt
     3721 ggaaaaaggg aaatcgaaga agttaaaatc cgttaaagag ttactaggga tcacaattat
     3781 ggaaagaagt tcctttgaaa aaaatccgat tgacttttta gaagctaaag gatataagga
     3841 agttaaaaaa gacttaatca ttaaactacc taaatatagt ctttttgagt tagaaaacgg
     3901 tcgtaaacgg atgctggcta gtgccggaga attacaaaaa ggaaatgagc tggctctgcc
     3961 aagcaaatat gtgaattttt tatatttagc tagtcattat gaaaagttga agggtagtcc
     4021 agaagataac gaacaaaaac aattgtttgt ggagcagcat aagcattatt tagatgagat
     4081 tattgagcaa atcagtgaat tttctaagcg tgttatttta gcagatgcca atttagataa
     4141 agttcttagt gcatataaca aacatagaga caaaccaata cgtgaacaag cagaaaatat
     4201 tattcattta tttacgttga cgaatcttgg agctcccgct gcttttaaat attttgatac
     4261 aacaattgat cgtaaacgat atacgtctac aaaagaagtt ttagatgcca ctcttatcca
     4321 tcaatccatc actggtcttt atgaaacacg cattgatttg agtcagctag gaggtgactg
     4381 agggcccgga tccggtaccg aattcaagct tgacctgtga agtgaaaaat ggcgcacatt
     4441 gtgcgacatt ttttttgtct gccgtttacc gctactgcgt cacggatctc cacgcgccct
     4501 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     4561 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     4621 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     4681 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     4741 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     4801 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     4861 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     4921 ttaacaaaat attaacgctt acaatttcag gtggcacttt tcggggaaat gtgcgcggaa
     4981 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     5041 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     5101 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     5161 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     5221 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     5281 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     5341 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     5401 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     5461 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     5521 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5581 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     5641 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttgatagact
     5701 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     5761 ttattgctga taaatctgga gccggtgagc gtggctctcg cggtatcatt gcagcactgg
     5821 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     5881 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtagg
     5941 aattaatgat gtctcgttta gataaaagta aagtgattaa cagcgcatta gagctgctta
     6001 atgaggtcgg aatcgaaggt ttaacaaccc gtaaactcgc ccagaagcta ggtgtagagc
     6061 agcctacatt gtattggcat gtaaaaaata agcgggcttt gctcgacgcc ttagccattg
     6121 agatgttaga taggcaccat actcactttt gccctttaga aggggaaagc tggcaagatt
     6181 ttttacgtaa taacgctaaa agttttagat gtgctttact aagtcatcgc gatggagcaa
     6241 aagtacattt aggtacacgg cctacagaaa aacagtatga aactctcgaa aatcaattag
     6301 cctttttatg ccaacaaggt ttttcactag agaatgcatt atatgcactc agcgcagtgg
     6361 ggcattttac tttaggttgc gtattggaag atcaagagca tcaagtcgct aaagaagaaa
     6421 gggaaacacc tactactgat agtatgccgc cattattacg acaagctatc gaattatttg
     6481 atcaccaagg tgcagagcca gccttcttat tcggccttga attgatcata tgcggattag
     6541 aaaaacaact taaatgtgaa agtgggtctt aaaagcagca taaccttttt ccgtgatggt
     6601 aacttcacta gtttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     6661 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaat tgtcgggaag
     6721 atgcgtgatc tgatccttca actcagcaaa agttcgattt attcaacaaa gccgccgtcc
     6781 cgtcaagtca gcgtaatgct ctgccagtgt tacaaccaat taaccaattc tgatttattt
     6841 gccgactacc ttggtgatct cgcctttcac gtagtggaca aattcttcca actgatctgc
     6901 gcgcgaggcc aagcgatctt cttcttgtcc aagataagcc tgtctagctt caagtatgac
     6961 gggctgatac tgggccggca ggcgctccat tgcccagtcg gcagcgacat ccttcggcgc
     7021 gattttgccg gttactgcgc tgtaccaaat gcgggacaac gtaagcacta catttcgctc
     7081 atcgccagcc cagtcgggcg gcgagttcca tagcgttaag gtttcattta gcgcctcaaa
     7141 tagatcctgt tcaggaaccg gatcaaagag ttcctccgcc gctggaccta ccaaggcaac
     7201 gctatgttct cttgcttttg tcagcaagat agccagatca atgtcgatcg tggctggctc
     7261 gaagatacct gcaagaatgt cattgcgctg ccattctcca aattgcagtt cgcgcttagc
     7321 tggataacgc cacggaatga tgtcgtcgtg cacaacaatg gtgacttcta cagcgcggag
     7381 aatctcgctc tctccagggg aagccgaagt ttccaaaagg tcgttgatca aagctcgccg
     7441 cgttgtttca tcaagcctta cggtcaccgt aaccagcaaa tcaatatcac tgtgtggctt
     7501 caggccgcca tccactgcgg agccgtacaa atgtacggcc agcaacgtcg gttcgagatg
     7561 gcgctcgatg acgccaacta cctctgatag ttgagtcgat acttcggcga tcaccgcttc
     7621 cctcatgatg tttaactttg ttttagggcg actgccctgc tgcgtaacat cgttgctgct
     7681 ccataacatc aaacatcgac ccacggcgta acgcgcttgc tgcttggatg cccgaggcat
     7741 agactgtacc ccaaaaaaac agtcataaca agccattaac accccttgta ttactgttta
     7801 tgtaagcaga cagttttatt gttcatgatg atatattttt atcttgtgca atgtaacatc
     7861 agagattttg agacacaacg tggctttcga cttccggcaa gctatacgcg ccctagaatt
     7921 gtcaatttta atcctctgtt tatcggcagt tcgtagagcg cgccgtgcgt cccgagcgat
     7981 actgagcgaa gcaagtgcgt cgagcagtgc ccgcttgttc ctgaaatgcc agtaaagcgc
     8041 tggctgctga acccccagcc ggaactgacc ccacaaggcc ctagcgtttg caatgcacca
     8101 ggtcatcatt gacccaggcg tgttccacca ggccgctgcc tcgcaactct tcgcaggctt
     8161 cgccgacctg ctcgcgccac ttcttcacgc gggtggaatc cgatccgcac atgaggcgga
     8221 aggtttccag cttgagcggg tacggctccc ggtgcgagct gaaatagtcg aacatccgtc
     8281 gggccgtcgg cgacagcttg cggtacttct cccatatgaa tttcgtgtag tggtcgccag
     8341 caaacagcac gacgatttcc tcgtcgatca ggacctggca acgggacgtt ttcttgccac
     8401 ggtccaggac gcggaagcgg tgcagcagcg acaccgattc caggtgccca acgcggtcgg
     8461 acgtgaagcc catcgccgtc gcctgtaggc gcgacaggca ttcctcggcc ttcgtgtaat
     8521 accggccatt gatcgaccag cccaggtcct ggcaaagctc gtagaacgtg aaggtgatcg
     8581 gctcgccgat aggggtgcgc ttcgcgtact ccaacacctg ctgccacacc agttcgtcat
     8641 cgtcggcccg cagctcgacg ccggtgtagg tgatcttcac gtccttgttg acgtggaaaa
     8701 tgaccttgtt ttgcagcgcc tcgcgcggga ttttcttgtt gcgcgtggtg aacagggcag
     8761 agcgggccgt gtcgtttggc atcgctcgca tcgtgtccgg ccacggcgca atatcgaaca
     8821 aggaaagctg catttccttg atctgctgct tcgtgtgttt cagcaacgcg gcctgcttgg
     8881 cctcgctgac ctgttttgcc aggtcctcgc cggcggtttt tcgcttcttg gtcgtcatag
     8941 ttcctcgcgt gtcgatggtc atcgacttcg ccaaacctgc cgcctcctgt tcgagacgac
     9001 gcgaacgctc cacggcggcc gatggcgcgg gcagggcagg gggagccagt tgcacgctgt
     9061 cgcgctcgat cttggccgta gcttgctgga ccatcgagcc gacggactgg aaggtttcgc
     9121 ggggcgcacg catgacggtg cggcttgcga tggtttcggc atcctcggcg gaaaaccccg
     9181 cgtcgatcag ttcttgcctg tatgccttcc ggtcaaacgt ccgattcatt caccctcctt
     9241 gcgggattgc cccgactcac gccggggcaa tgtgccctta ttcctgattt gacccgcctg
     9301 gtgccttggt gtccagataa tccaccttat cggcaatgaa gtcggtcccg tagaccgtct
     9361 ggccgtcctt ctcgtacttg gtattccgaa tcttgccctg cacgaatacc agctccgcga
     9421 agtcgctctt cttgatggag cgcatgggga cgtgcttggc aatcacgcgc accccccggc
     9481 cgttttagcg gctaaaaaag tcatggctct gccctcgggc ggaccacgcc catcatgacc
     9541 ttgccaagct cgtcctgctt ctcttcgatc ttcgccagca gggcgaggat cgtggcatca
     9601 ccgaaccgcg ccgtgcgcgg gtcgtcggtg agccagagtt tcagcaggcc gcccaggcgg
     9661 cccaggtcgc cattgatgcg ggccagctcg cggacgtgct catagtccac gacgcccgtg
     9721 attttgtagc cctggccgac ggccagcagg taggcctaca ggctcatgcc ggccgccgcc
     9781 gccttttcct caatcgctct tcgttcgtct ggaaggcagt acaccttgat aggtgggctg
     9841 cccttcctgg ttggcttggt ttcatcagcc atccgcttgc cctcatctgt tacgccggcg
     9901 gtagccggcc agcctcgcag agcaggattc ccgttgagca ccgccaggtg cgaataaggg
     9961 acagtgaaga aggaacaccc gctcgcgggt gggcctactt cacctatcct gcccggctga
    10021 cgccgttgga tacaccaagg aaagtctaca cgaacccttt ggcaaaatcc tgtatatcgt
    10081 gcgaaaaagg atggatatac cgaaaaaatc gctataatga ccccgaagca gggttatgca
    10141 gcggaaaaga tccgtcgacc ctttccgacg ctcaccgggc tggttgccct cgccgctggg
    10201 ctggcggccg tctatggccc tgcaaacgcg ccagaaacgc cgtcgaagcc gtgtgcgaga
    10261 caccgcggcc gccggcgttg tggatacctc gcggaaaact tggccctcac tgacagatga
    10321 ggggcggacg ttgacacttg aggggccgac tcacccggcg cggcgttgac agatgagggg
    10381 caggctcgat ttcggccggc gacgtggagc tggccagcct cgcaaatcgg cgaaaacgcc
    10441 tgattttacg cgagtttccc acagatgatg tggacaagcc tggggataag tgccctgcgg
    10501 tattgacact tgaggggcgc gactactgac agatgagggg cgcgatcctt gacacttgag
    10561 gggcagagtg ctgacagatg aggggcgcac ctattgacat ttgaggggct gtccacaggc
    10621 agaaaatcca gcatttgcaa gggtttccgc ccgtttttcg gccaccgcta acctgtcttt
    10681 taacctgctt ttaaaccaat atttataaac cttgttttta accagggctg cgccctgtgc
    10741 gcgtgaccgc gcacgccgaa ggggggtgcc cccccttctc gaaccctccc ggcccgctaa
    10801 cgcgggcctc ccatcccccc aggggctgcg cccctcggcc gcgaacggcc tcaccccaaa
    10861 aatggcagcc aagctgaccc gctagggacg tgaagtcgat tccttcgatg gttagcaatc
    10921 aaagactcag agtgctgtct ggagcgtgaa tctaacggta cgtatctcga ttgctcggtc
    10981 gctattcgca ctctgcgaaa gttcgtaccg ctcattcact aggttgcgaa tcatgaccaa
    11041 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
    11101 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
    11161 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
    11221 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
    11281 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
    11341 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
    11401 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
    11461 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
    11521 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
    11581 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
    11641 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaa
//
