LOCUS       sequence-181033-       12171 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genome engineering of virulent phages.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12171)
  AUTHORS   Lemay ML, Tremblay DM, Moineau S
  TITLE     Genome Engineering of Virulent Lactococcal Phages Using CRISPR-Cas9.
  JOURNAL   ACS Synth Biol. 2017 Mar 30. doi: 10.1021/acssynbio.6b00388.
  PUBMED    28324650
REFERENCE   2  (bases 1 to 12171)
  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: "ACS Synth 
            Biol. 2017 Mar 30. doi: 10.1021/acssynbio.6b00388."
COMMENT     Sequence Label: pL2Cas9
FEATURES             Location/Qualifiers
     source          1..12171
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     162..181
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(248..350)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(742..759)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(876..1421)
                     /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(956..975)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6272..6291
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     repeat_region   6720..6755
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     repeat_region   6786..6821
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     misc_feature    complement(6822..6953)
                     /label=crRNA leader
                     /note="crRNA leader sequence for the Streptococcus pyogenes
                     CRISPR/Cas system"
     CDS             complement(6974..11080)
                     /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             complement(6977..11080)
                     /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_RNA        11376..11454
                     /label=tracrRNA
                     /note="trans-activating CRISPR RNA for the Streptococcus 
                     pyogenes CRISPR/Cas9 system"
     promoter        complement(join(11772..11777,11778..11794,11795..11800))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     11889..11906
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     11890..11906
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11985..12001)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11985..12001)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11998..12020)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12009..12025
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(12033..12039,12040..12057,12058..12063))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12078..12099
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cgcgcgggga gaggcggttt gcgtattgga attccggatg agcattcatc aggcgggcaa
       61 gaatgtgaat aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg
      121 ccgtaatatc cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct
      181 caaaatgttc tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt
      241 tctccatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta
      301 gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgccgatca acgtctcatt
      361 ttcgccaaaa gttggcccag ggcttcccgg tatcaacagg gacaccagga tttatttatt
      421 ctgcgaagtg atcttccgtc acaggtattt attcggcgca aagtgcgtcg ggtgatgctg
      481 ccaacttact gatttagtgt atgatggtgt ttttgaggtg ctccagtggc ttctgtttct
      541 atcagctgtc cctcctgttc agctactgac ggggtggtgc gtaacggcaa aagcaccgcc
      601 ggacatcagc gctagcggag tgtatactgg cttactatgt tggcactgat gagggtgtca
      661 gtgaagtgct tcatgtggca ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg
      721 atacaggata tattccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg
      781 gcgagcggaa atggcttacg aacggggcgg agatttcctg gaagatgcca ggaagatact
      841 taacagggaa gtgagagggc cgcggcaaag ccgtttttcc ataggctccg cccccctgac
      901 aagcatcacg aaatctgacg ctcaaatcag tggtggcgaa acccgacagg actataaaga
      961 taccaggcgt ttccccctgg cggctccctc gtgcgctctc ctgttcctgc ctttcggttt
     1021 accggtgtca ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga cactcagttc
     1081 cgggtaggca gttcgctcca agctggactg tatgcacgaa ccccccgttc agtccgaccg
     1141 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gaaagacatg caaaagcacc
     1201 actggcagca gccactggta attgatttag aggagttagt cttgaagtca tgcgccggtt
     1261 aaggctaaac tgaaaggaca agttttggtg actgcgctcc tccaagccag ttacctcggt
     1321 tcaaagagtt ggtagctcag agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt
     1381 tttcagagca agagattacg cgcagaccaa aacgatctca agaagatcat cttattaatc
     1441 agataaaata tttctagact cgagagctct cgagtctaga atcgatacga ttttgaagtg
     1501 gcaacagata aaaaaaagca gtttaaaatt gttgctgaac ttttaaaaca agcaaataca
     1561 atcattgtcg caacagatag cgacagagaa ggcgaaaaca ttgcctggtc gatcattcat
     1621 aaagcaaatg ccttttctaa agataaaacg tataaaagac tatggatcaa tagtttagaa
     1681 aaagatgtga tccgtagcgg ttttcaaaat ttgcaaccag gaatgaatta ctatcccttt
     1741 tatcaagaag cgcacaaaaa gaaaaacgaa atgatacacc aatcagtgca aaaaaagata
     1801 taatgggaga taagacggtt cgtgttcgtg ctgacttgca ccatatcata aaaatcgaaa
     1861 cagcaaagaa tggcggaaac gtaaaagaag ttatggaaat aagacttaga agcaaactta
     1921 agagtgtgtt gatagtgcag tatcttaaaa ttttgtataa taggaattga agttaaatta
     1981 gatgctaaaa atttgtaatt aagaaggagt gattacatga acaaaaatat aaaatattct
     2041 caaaactttt taacgagtga aaaagtactc aaccaaataa taaaacaatt gaatttaaaa
     2101 gaaaccgata ccgtttacga aattggaaca ggtaaagggc atttaacgac gaaactggct
     2161 aaaataagta aacaggtaac gtctattgaa ttagacagtc atctattcaa cttatcgtca
     2221 gaaaaattaa aactgaatac tcgtgtcact ttaattcacc aagatattct acagtttcaa
     2281 ttccctaaca aacagaggta taaaattgtt gggagtattc cttaccattt aagcacacaa
     2341 attattaaaa aagtggtttt tgaaagccat gcgtctgaca tctatctgat tgttgaagaa
     2401 ggattctaca agcgtacctt ggatattcac cgaacactag ggttgctctt gcacactcaa
     2461 gtctcgattc agcaattgct taagctgcca gcggaatgct ttcatcctaa accaaaagta
     2521 aacagtgtct taataaaact tacccgccat accacagatg ttccagataa atattggaag
     2581 ctatatacgt actttgtttc aaaatgggtc aatcgagaat atcgtcaact gtttactaaa
     2641 aatcagtttc atcaagcaat gaaacacgcc aaagtaaaca atttaagtac cgttacttat
     2701 gagcaagtat tgtctatttt taatagttat ctattattta acgggaggaa ataattctat
     2761 gagtcgcttt tgtaaatttg gaaagttaca cgttactaaa gggaatgtag ataaattatt
     2821 aggtatacta ctgacagctt ccaaggagct aaagaggtcc ctagcgctta gaatcgcttt
     2881 aggaaacacg atccagtcca ataatcgtcg ataaaaactt ttgaaaaagg ttggtgaaat
     2941 tacctacttt tggaataatc acaaatcaca agtgattaat cacaaatcac aagtgattaa
     3001 tcacttgttt attaagatat taaaagctat aatttaaata aagcgtgaat tttattacac
     3061 aaaaagaggg gggagaaact tggaactagc atttagagaa agcttaaaaa agatgagagg
     3121 taccaaatca aaagaaaaat tctcccaaga attagaaatg agtagatcaa attattcacg
     3181 aatagaatca ggaaaatcag atccaaccat aaaaacacta gaacaaattg caaagttaac
     3241 taactcaacg ctagtagtgg atttaatccc aaatgagcca acagaaccag aaccagaaac
     3301 agaatcagaa caagtaacat tggatttaga aatggaagaa gaaaaaagca atgacttcgt
     3361 gtgaataatg cacgaaatcg ttgcttattt tttttaaaag cggtatacta gatataacga
     3421 aacaacgaac tgaatagaaa cgaaaaaaga gccatgacac atttataaaa tgtttgacga
     3481 cattttataa atgcatagcc cgataagatt gccaaaccaa cgcttatcag ttagtcagat
     3541 gaactcttcc ctcgtaagaa gttatttaat taactttgtt tgaagacggt atataaccgt
     3601 actatcatta tatagggaaa tcagagagtt ttcaagtatc taagctactg aatttaagaa
     3661 ttgttaagca atcaatcgga aatcgtttga ttgctttttt tgtattcatt tatagaaggt
     3721 ggagtttgta tgaatcatga tgaatgtaaa acttatataa aaaatagttt attggagata
     3781 agaaaattag caaatatcta tacactagaa acgtttaaga aagagttaga aaagagaaat
     3841 atctacttag aaacaaaatc agataagtat ttttcttcgg agggggaaga ttatatatat
     3901 aagttaatag aaaataacaa aataatttat tcgattagtg gaaaaaaatt gacttataaa
     3961 ggaaaaaaat ctttttcaaa acatgcaata ttgaaacagt tgaatgaaaa agcaaaccaa
     4021 gttaattaaa caacctattt tataggattt ataggaaagg agaacagctg aatgaatatc
     4081 ccttttgttg tagaaactgt gcttcatgac ggcttgttaa agtacaaatt taaaaatagt
     4141 aaaattcgct caatcactac caagccaggt aaaagcaaag gggctatttt tgcgtatcgc
     4201 tcaaaatcaa gcatgattgg cggtcgtggt gttgttctga cttccgagga agcgattcaa
     4261 gaaaatcaag atacatttac acattggaca cccaacgttt atcgttatgg aacgtatgca
     4321 gacgaaaacc gttcatacac gaaaggacat tctgaaaaca atttaagaca aatcaatacc
     4381 ttctttattg attttgatat tcacacggca aaagaaacta tttcagcaag cgatatttta
     4441 acaaccgcta ttgatttagg ttttatgcct actatgatta tcaaatctga taaaggttat
     4501 caagcatatt ttgttttaga aacgccagtc tatgtgactt caaaatcaga atttaaatct
     4561 gtcaaagcag ccaaaataat ttcgcaaaat atccgagaat attttggaaa gtctttgcca
     4621 gttgatctaa cgtgtaatca ttttggtatt gctcgcatac caagaacgga caatgtagaa
     4681 ttttttgatc ctaattaccg ttattctttc aaagaatggc aagattggtc tttcaaacaa
     4741 acagataata agggctttac tcgttcaagt ctaacggttt taagcggtac agaaggcaaa
     4801 aaacaagtag atgaaccctg gtttaatctc ttattgcacg aaacgaaatt ttcaggagaa
     4861 aagggtttaa tagggcgtaa taacgtcatg tttaccctct ctttagccta ctttagttca
     4921 ggctattcaa tcgaaacgtg cgaatataat atgtttgagt ttaataatcg attagatcaa
     4981 cccttagaag aaaaagaagt aatcaaaatt gttagaagtg cctattcaga aaactatcaa
     5041 ggggctaata gggaatacat taccattctt tgcaaagctt gggtatcaag tgatttaacc
     5101 agtaaagatt tatttgtccg tcaagggtgg tttaaattca agaaaaaaag aagcgaacgt
     5161 caacgtgttc atttgtcaga atggaaagaa gatttaatgg cttatattag cgaaaaaagc
     5221 gatgtataca agccttattt agtgacgacc aaaaaagaga ttagagaagt gctaggcatt
     5281 cctgaacgga cattagataa attgctgaag gtactgaagg cgaatcagga aattttcttt
     5341 aagattaaac caggaagaaa tggtggcatt caacttgcta gtgttaaatc attgttgcta
     5401 tcgatcatta aagtaaaaaa agaagaaaaa gaaagctata taaaggcgct gacaaattct
     5461 tttgacttag agcatacatt cattcaagag actttaaaca agctagcaga acgccctaaa
     5521 acggacacac aactcgattt gtttagctat gatacaggct gaaaataaaa cccgcactat
     5581 gccattacat ttatatctat gatacgtgtt tgttttttct ttgctgttta gcgaatgatt
     5641 agcagaaata tacagagtaa gattttaatt aattattagg gggagaagga gagagtagcc
     5701 cgaaaacttt tagttggctt ggactgaacg aagtgaggga aaggctacta aaacgtcgag
     5761 gggcagtgag agcgaagcga acacttgatt ttttaatttt ctatctttta taggtcatta
     5821 gagtatactt atttgtccta taaactattt agcagcataa tagatttatt gaataggtca
     5881 tttaagttga gcatattaga ggaggaaaat cttggagaaa tatttgaaga acccgattac
     5941 atggattgga ttagttcttg tggttacgtg gtttttaact aaaagtagtg aatttttgat
     6001 ttttggtgtg tgtgtcttgt tgttagtatt tgctagtcaa agtgattaaa tagaattcat
     6061 tcgccattca ggctgcgcgc tctcccttat gcgactcctg cattaggaag cagcccagta
     6121 gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc
     6181 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
     6241 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
     6301 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atccacagga
     6361 cgggtgtggt cgccatgatc gcgtagtcga tagtggctcc aagtagcgaa gcgagcagga
     6421 ctgggcggcg gccaaagcgg tcggacagtg ctccgagaac gggtgcgcat agaaattgca
     6481 tcaacgcata tagcgctagc agcacgccat agtgactggc gatgctgtcg gaatggacga
     6541 tcacactact cttcttttgc ctattataac atgaaaacgc tttaacagaa agaataggaa
     6601 ggtatccgac tgctggtatt aaccctcttt ctcaagttat catcggcaat gttgaatgga
     6661 gtccattcaa aacagcatag ctctaaaaca tggaactcaa caagtctcag tgtgctgaag
     6721 ttttgggacc attcaaaaca gcatagctct aaaacgagac ctttgagctt ccgagactgg
     6781 tctcagtttt gggaccattc aaaacagcat agctctaaaa cctcgtagac tatttttgtc
     6841 taaaaaattt cgtaatcgca ctatttgtct cagctagact tcagtcttga aaagcccctg
     6901 tattactgca tttattaaga gtattatacc atatttttag ttattaagaa ataatcttca
     6961 tctaaaatat acttcagtca cctcctagct gactcaaatc aatgcgtgtt tcataaagac
     7021 cagtgatgga ttgatggata agagtggcat ctaaaacttc ttttgtagac gtatatcgtt
     7081 tacgatcaat tgttgtatca aaatatttaa aagcagcggg agctccaaga ttcgtcaacg
     7141 taaataaatg aataatattt tctgcttgtt cacgtattgg tttgtctcta tgtttgttat
     7201 atgcactaag aactttatct aaattggcat ctgctaaaat aacacgctta gaaaattcac
     7261 tgatttgctc aataatctca tctaaataat gcttatgctg ctccacaaac aattgttttt
     7321 gttcgttatc ttctggacta cccttcaact tttcataatg actagctaaa tataaaaaat
     7381 tcacatattt gcttggcaga gccagctcat ttcctttttg taattctccg gcactagcca
     7441 gcatccgttt acgaccgttt tctaactcaa aaagactata tttaggtagt ttaatgatta
     7501 agtctttttt aacttcctta tatcctttag cttctaaaaa gtcaatcgga tttttttcaa
     7561 aggaacttct ttccataatt gtgatcccta gtaactcttt aacggatttt aacttcttcg
     7621 atttcccttt ttccacctta gcaaccacta ggactgaata agctaccgtt ggactatcaa
     7681 aaccaccata tttttttgga tcccagtctt ttttacgagc aataagcttg tccgaatttc
     7741 tttttggtaa aattgactcc ttggagaatc cgcctgtctg tacttctgtt ttcttgacaa
     7801 tattgacttg gggcatggac aatactttgc gcactgtggc aaaatctcgc cctttatccc
     7861 agacaatttc tccagtttcc ccattagttt cgattagagg gcgtttgcga atctctccat
     7921 ttgcaagtgt aatttctgtt ttgaagaagt tcatgatatt agagtaaaag aaatattttg
     7981 cggttgcttt gcctatttct tgctcagact tagcaatcat tttacgaaca tcataaactt
     8041 tataatcacc atagacaaac tccgattcaa gttttggata tttcttaatc aaagcagttc
     8101 caacgacggc atttagatac gcatcatggg catgatggta attgttaatc tcacgtactt
     8161 tatagaattg gaaatctttt cggaagtcag aaactaattt agattttaag gtaatcactt
     8221 taacctctcg aataagttta tcattttcat cgtatttagt attcatgcga ctatccaaaa
     8281 tttgtgccac atgcttagtg atttggcgag tttcaaccaa ttggcgtttg ataaaaccag
     8341 ctttatcaag ttcactcaaa cctccacgtt cagctttcgt taaattatca aacttacgtt
     8401 gagtgattaa cttggcgttt agaagttgtc tccaatagtt tttcatcttt ttgactactt
     8461 cttcacttgg aacgttatcc gatttaccac gatttttatc agaacgcgtt aagaccttat
     8521 tgtctattga atcgtcttta aggaaacttt gtggaacaat gtgatcgaca tcataatcac
     8581 ttaaacgatt aatatctaat tcttggtcca catacatgtc tcttccattt tggagataat
     8641 agagatagag cttttcattt tgcaattgag tattttcaac aggatgctct ttaagaatct
     8701 gacttcctaa ttctttgata ccttcttcga ttcgtttcat acgctctcgc gaatttttct
     8761 ggcccttttg agttgtctga ttttcacgtg ccatttcaat aacgatattt tctggcttat
     8821 gccgccccat tactttgacc aattcatcaa caacttttac agtctgtaaa ataccttttt
     8881 taatagcagg gctaccagct aaatttgcaa tatgttcatg taaactatcg ccttgtccag
     8941 acacttgtgc tttttgaatg tcttctttaa atgtcaaact atcatcatgg atcagctgca
     9001 taaaattgcg attggcaaaa ccatctgatt tcaaaaaatc taatattgtt ttgccagatt
     9061 gcttatccct aataccatta atcaattttc gagacaaacg tccccaacca gtataacggc
     9121 gacgtttaag ctgtttcatc accttatcat caaagaggtg agcatatgtt ttaagtcttt
     9181 cctcaatcat ctccctatct tcaaataagg tcaatgttaa aacaatatcc tctaagatat
     9241 cttcattttc ttcattatcc aaaaaatctt tatctttaat aatttttagc aaatcatggt
     9301 aggtacctaa tgaagcatta aatctatctt caactcctga aatttcaaca ctatcaaaac
     9361 attctatttt tttgaaataa tcttctttta attgcttaac ggttactttt cgatttgttt
     9421 tgaagagtaa atcaacaatg gctttcttct gttcacctga aagaaatgct ggttttcgca
     9481 ttccttcagt aacatatttg acctttgtca attcgttata aaccgtaaaa tactcataaa
     9541 gcaaactatg ttttggtagt actttttcat ttggaagatt tttatcaaag tttgtcatgc
     9601 gttcaataaa tgattgagct gaagcacctt tatcgacaac ttcttcaaaa ttccatgggg
     9661 taattgtttc ttcagacttc cgagtcatcc atgcaaaacg actattgcca cgcgccaatg
     9721 gaccaacata ataaggaatt cgaaaagtca agattttttc aatcttctca cgattgtctt
     9781 ttaaaaatgg ataaaagtct tcttgtcttc tcaaaatagc atgcagctca cccaagtgaa
     9841 tttgatgggg aatagagccg ttgtcaaagg tccgttgctt gcgcagcaaa tcttcacgat
     9901 ttagtttcac caataattcc tcagtaccat ccattttttc taaaattggt ttgataaatt
     9961 tataaaattc ttcttggcta gctcccccat caatataacc tgcatatccg ttttttgatt
    10021 gatcaaaaaa gatttcttta tacttttctg gaagttgttg tcgaactaaa gcttttaaaa
    10081 gagtcaagtc ttgatgatgt tcatcgtagc gtttaatcat tgaagctgat aggggagcct
    10141 tagttatttc agtatttact cttaggatat ctgaaagtaa aatagcatct gataaattct
    10201 tagctgccaa aaacaaatca gcatattgat ctccaatttg cgccaataaa ttatctaaat
    10261 catcatcgta agtatctttt gaaagctgta atttagcatc ttctgccaaa tcaaaatttg
    10321 atttaaaatt aggggtcaaa cccaatgaca aagcaatgag attcccaaat aagccatttt
    10381 tcttctcacc ggggagctga gcaatgagat tttctaatcg tcttgattta ctcaatcgtg
    10441 cagaaagaat cgctttagca tctactccac ttgcgttaat agggttttct tcaaataatt
    10501 gattgtaggt ttgtaccaac tggataaata gtttgtccac atcactatta tcaggattta
    10561 aatctccctc aatcaaaaaa tgaccacgaa acttaatcat atgcgctaag gccaaataga
    10621 ttaagcgcaa atccgcttta tcagtagaat ctaccaattt ttttcgcaga tgatagatag
    10681 ttggatattt ctcatgataa gcaacttcat ctactatatt tccaaaaata ggatgacgtt
    10741 catgcttctt gtcttcttcc accaaaaaag actcttcaag tcgatgaaag aaactatcat
    10801 ctactttcgc catctcattt gaaaaaatct cctgtagata acaaatacga ttcttccgac
    10861 gtgtatacct tctacgagct gtccgtttga gacgagtcgc ttccgctgtc tctccactgt
    10921 caaataaaag agcccctata agattttttt tgatactgtg gcggtctgta tttcccagaa
    10981 ccttgaactt tttagacgga accttatatt catcagtgat caccgcccat ccgacgctat
    11041 ttgtgccgat atctaagcct attgagtatt tcttatccat ttttgcctcc taaaataaaa
    11101 agtttaaatt aaatccataa tgagtttgat gatttcaata atagttttaa tgacctccga
    11161 aattagttta atatgcttta atttttcttt ttcaaaatat ctcttcaaaa aatattaccc
    11221 aatacttaat aataaataga ttataacaca aaattctttt aaaaagtagt ttattttgtt
    11281 atcattctat agtattaagt attgttttat ggctgataaa tttctttgaa tttctccttg
    11341 attatttgtt ataaaagtta taaaataatc ttgttggaac cattcaaaac agcatagcaa
    11401 gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt
    11461 tgatacttct attctactct gactgcaaac caaaaaaaca agcgctttca aaacgcttgt
    11521 tttatcattt ttagggaaat taatctctta atccttttat cattctacat ttaggcgctg
    11581 ccatcttgct aaacctacta agctccacag gatgatttcg taatcccgca agaggcccgg
    11641 cagtaccggc ataaccaagc ctatgcctac agcatccagg gtgacggtgc cgaggatgac
    11701 gatgagcgca ttgttagatt tcatacacgg tgcctgactg cgttagcaat ttaactgtga
    11761 taaactaccg cattaaagct tatcgatgat aagctgtcaa acatgagaat tacaacttat
    11821 atcgtatggg gctgacttca ggtgctacat ttgaagagat aaattgcact gaaatccagt
    11881 cacgacgttg taaaacgacg gccagtgaat tcccggggat ccgtcgacct gcagccaagc
    11941 tttcgcgagc tcgagatcta gatatcgatg aattcgtaat catggtcata gctgtttcct
    12001 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
    12061 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
    12121 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca a
//
