LOCUS       sequence-197035-       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     146..165
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(232..334)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(726..743)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(860..1405)
                     /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(940..959)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6256..6275
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     repeat_region   6704..6739
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     repeat_region   6770..6805
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     misc_feature    complement(6806..6937)
                     /label=crRNA leader
                     /note="crRNA leader sequence for the Streptococcus pyogenes
                     CRISPR/Cas system"
     CDS             complement(6958..11064)
                     /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(6961..11064)
                     /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        11360..11438
                     /label=tracrRNA
                     /note="trans-activating CRISPR RNA for the Streptococcus 
                     pyogenes CRISPR/Cas9 system"
     promoter        complement(join(11756..11761,11762..11778,11779..11784))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     11873..11890
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     11874..11890
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11969..11985)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11969..11985)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11982..12004)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11993..12009
                     /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(12017..12023,12024..12041,12042..12047))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12062..12083
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gtttgcgtat tggaattccg gatgagcatt catcaggcgg gcaagaatgt gaataaaggc
       61 cggataaaac ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa tatccagctg
      121 aacggtctgg ttataggtac attgagcaac tgactgaaat gcctcaaaat gttctttacg
      181 atgccattgg gatatatcaa cggtggtata tccagtgatt tttttctcca ttttagcttc
      241 cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt
      301 atggtgaaag ttggaacctc ttacgtgccg atcaacgtct cattttcgcc aaaagttggc
      361 ccagggcttc ccggtatcaa cagggacacc aggatttatt tattctgcga agtgatcttc
      421 cgtcacaggt atttattcgg cgcaaagtgc gtcgggtgat gctgccaact tactgattta
      481 gtgtatgatg gtgtttttga ggtgctccag tggcttctgt ttctatcagc tgtccctcct
      541 gttcagctac tgacggggtg gtgcgtaacg gcaaaagcac cgccggacat cagcgctagc
      601 ggagtgtata ctggcttact atgttggcac tgatgagggt gtcagtgaag tgcttcatgt
      661 ggcaggagaa aaaaggctgc accggtgcgt cagcagaata tgtgatacag gatatattcc
      721 gcttcctcgc tcactgactc gctacgctcg gtcgttcgac tgcggcgagc ggaaatggct
      781 tacgaacggg gcggagattt cctggaagat gccaggaaga tacttaacag ggaagtgaga
      841 gggccgcggc aaagccgttt ttccataggc tccgcccccc tgacaagcat cacgaaatct
      901 gacgctcaaa tcagtggtgg cgaaacccga caggactata aagataccag gcgtttcccc
      961 ctggcggctc cctcgtgcgc tctcctgttc ctgcctttcg gtttaccggt gtcattccgc
     1021 tgttatggcc gcgtttgtct cattccacgc ctgacactca gttccgggta ggcagttcgc
     1081 tccaagctgg actgtatgca cgaacccccc gttcagtccg accgctgcgc cttatccggt
     1141 aactatcgtc ttgagtccaa cccggaaaga catgcaaaag caccactggc agcagccact
     1201 ggtaattgat ttagaggagt tagtcttgaa gtcatgcgcc ggttaaggct aaactgaaag
     1261 gacaagtttt ggtgactgcg ctcctccaag ccagttacct cggttcaaag agttggtagc
     1321 tcagagaacc ttcgaaaaac cgccctgcaa ggcggttttt tcgttttcag agcaagagat
     1381 tacgcgcaga ccaaaacgat ctcaagaaga tcatcttatt aatcagataa aatatttcta
     1441 gactcgagag ctctcgagtc tagaatcgat acgattttga agtggcaaca gataaaaaaa
     1501 agcagtttaa aattgttgct gaacttttaa aacaagcaaa tacaatcatt gtcgcaacag
     1561 atagcgacag agaaggcgaa aacattgcct ggtcgatcat tcataaagca aatgcctttt
     1621 ctaaagataa aacgtataaa agactatgga tcaatagttt agaaaaagat gtgatccgta
     1681 gcggttttca aaatttgcaa ccaggaatga attactatcc cttttatcaa gaagcgcaca
     1741 aaaagaaaaa cgaaatgata caccaatcag tgcaaaaaaa gatataatgg gagataagac
     1801 ggttcgtgtt cgtgctgact tgcaccatat cataaaaatc gaaacagcaa agaatggcgg
     1861 aaacgtaaaa gaagttatgg aaataagact tagaagcaaa cttaagagtg tgttgatagt
     1921 gcagtatctt aaaattttgt ataataggaa ttgaagttaa attagatgct aaaaatttgt
     1981 aattaagaag gagtgattac atgaacaaaa atataaaata ttctcaaaac tttttaacga
     2041 gtgaaaaagt actcaaccaa ataataaaac aattgaattt aaaagaaacc gataccgttt
     2101 acgaaattgg aacaggtaaa gggcatttaa cgacgaaact ggctaaaata agtaaacagg
     2161 taacgtctat tgaattagac agtcatctat tcaacttatc gtcagaaaaa ttaaaactga
     2221 atactcgtgt cactttaatt caccaagata ttctacagtt tcaattccct aacaaacaga
     2281 ggtataaaat tgttgggagt attccttacc atttaagcac acaaattatt aaaaaagtgg
     2341 tttttgaaag ccatgcgtct gacatctatc tgattgttga agaaggattc tacaagcgta
     2401 ccttggatat tcaccgaaca ctagggttgc tcttgcacac tcaagtctcg attcagcaat
     2461 tgcttaagct gccagcggaa tgctttcatc ctaaaccaaa agtaaacagt gtcttaataa
     2521 aacttacccg ccataccaca gatgttccag ataaatattg gaagctatat acgtactttg
     2581 tttcaaaatg ggtcaatcga gaatatcgtc aactgtttac taaaaatcag tttcatcaag
     2641 caatgaaaca cgccaaagta aacaatttaa gtaccgttac ttatgagcaa gtattgtcta
     2701 tttttaatag ttatctatta tttaacggga ggaaataatt ctatgagtcg cttttgtaaa
     2761 tttggaaagt tacacgttac taaagggaat gtagataaat tattaggtat actactgaca
     2821 gcttccaagg agctaaagag gtccctagcg cttagaatcg ctttaggaaa cacgatccag
     2881 tccaataatc gtcgataaaa acttttgaaa aaggttggtg aaattaccta cttttggaat
     2941 aatcacaaat cacaagtgat taatcacaaa tcacaagtga ttaatcactt gtttattaag
     3001 atattaaaag ctataattta aataaagcgt gaattttatt acacaaaaag aggggggaga
     3061 aacttggaac tagcatttag agaaagctta aaaaagatga gaggtaccaa atcaaaagaa
     3121 aaattctccc aagaattaga aatgagtaga tcaaattatt cacgaataga atcaggaaaa
     3181 tcagatccaa ccataaaaac actagaacaa attgcaaagt taactaactc aacgctagta
     3241 gtggatttaa tcccaaatga gccaacagaa ccagaaccag aaacagaatc agaacaagta
     3301 acattggatt tagaaatgga agaagaaaaa agcaatgact tcgtgtgaat aatgcacgaa
     3361 atcgttgctt atttttttta aaagcggtat actagatata acgaaacaac gaactgaata
     3421 gaaacgaaaa aagagccatg acacatttat aaaatgtttg acgacatttt ataaatgcat
     3481 agcccgataa gattgccaaa ccaacgctta tcagttagtc agatgaactc ttccctcgta
     3541 agaagttatt taattaactt tgtttgaaga cggtatataa ccgtactatc attatatagg
     3601 gaaatcagag agttttcaag tatctaagct actgaattta agaattgtta agcaatcaat
     3661 cggaaatcgt ttgattgctt tttttgtatt catttataga aggtggagtt tgtatgaatc
     3721 atgatgaatg taaaacttat ataaaaaata gtttattgga gataagaaaa ttagcaaata
     3781 tctatacact agaaacgttt aagaaagagt tagaaaagag aaatatctac ttagaaacaa
     3841 aatcagataa gtatttttct tcggaggggg aagattatat atataagtta atagaaaata
     3901 acaaaataat ttattcgatt agtggaaaaa aattgactta taaaggaaaa aaatcttttt
     3961 caaaacatgc aatattgaaa cagttgaatg aaaaagcaaa ccaagttaat taaacaacct
     4021 attttatagg atttatagga aaggagaaca gctgaatgaa tatccctttt gttgtagaaa
     4081 ctgtgcttca tgacggcttg ttaaagtaca aatttaaaaa tagtaaaatt cgctcaatca
     4141 ctaccaagcc aggtaaaagc aaaggggcta tttttgcgta tcgctcaaaa tcaagcatga
     4201 ttggcggtcg tggtgttgtt ctgacttccg aggaagcgat tcaagaaaat caagatacat
     4261 ttacacattg gacacccaac gtttatcgtt atggaacgta tgcagacgaa aaccgttcat
     4321 acacgaaagg acattctgaa aacaatttaa gacaaatcaa taccttcttt attgattttg
     4381 atattcacac ggcaaaagaa actatttcag caagcgatat tttaacaacc gctattgatt
     4441 taggttttat gcctactatg attatcaaat ctgataaagg ttatcaagca tattttgttt
     4501 tagaaacgcc agtctatgtg acttcaaaat cagaatttaa atctgtcaaa gcagccaaaa
     4561 taatttcgca aaatatccga gaatattttg gaaagtcttt gccagttgat ctaacgtgta
     4621 atcattttgg tattgctcgc ataccaagaa cggacaatgt agaatttttt gatcctaatt
     4681 accgttattc tttcaaagaa tggcaagatt ggtctttcaa acaaacagat aataagggct
     4741 ttactcgttc aagtctaacg gttttaagcg gtacagaagg caaaaaacaa gtagatgaac
     4801 cctggtttaa tctcttattg cacgaaacga aattttcagg agaaaagggt ttaatagggc
     4861 gtaataacgt catgtttacc ctctctttag cctactttag ttcaggctat tcaatcgaaa
     4921 cgtgcgaata taatatgttt gagtttaata atcgattaga tcaaccctta gaagaaaaag
     4981 aagtaatcaa aattgttaga agtgcctatt cagaaaacta tcaaggggct aatagggaat
     5041 acattaccat tctttgcaaa gcttgggtat caagtgattt aaccagtaaa gatttatttg
     5101 tccgtcaagg gtggtttaaa ttcaagaaaa aaagaagcga acgtcaacgt gttcatttgt
     5161 cagaatggaa agaagattta atggcttata ttagcgaaaa aagcgatgta tacaagcctt
     5221 atttagtgac gaccaaaaaa gagattagag aagtgctagg cattcctgaa cggacattag
     5281 ataaattgct gaaggtactg aaggcgaatc aggaaatttt ctttaagatt aaaccaggaa
     5341 gaaatggtgg cattcaactt gctagtgtta aatcattgtt gctatcgatc attaaagtaa
     5401 aaaaagaaga aaaagaaagc tatataaagg cgctgacaaa ttcttttgac ttagagcata
     5461 cattcattca agagacttta aacaagctag cagaacgccc taaaacggac acacaactcg
     5521 atttgtttag ctatgataca ggctgaaaat aaaacccgca ctatgccatt acatttatat
     5581 ctatgatacg tgtttgtttt ttctttgctg tttagcgaat gattagcaga aatatacaga
     5641 gtaagatttt aattaattat tagggggaga aggagagagt agcccgaaaa cttttagttg
     5701 gcttggactg aacgaagtga gggaaaggct actaaaacgt cgaggggcag tgagagcgaa
     5761 gcgaacactt gattttttaa ttttctatct tttataggtc attagagtat acttatttgt
     5821 cctataaact atttagcagc ataatagatt tattgaatag gtcatttaag ttgagcatat
     5881 tagaggagga aaatcttgga gaaatatttg aagaacccga ttacatggat tggattagtt
     5941 cttgtggtta cgtggttttt aactaaaagt agtgaatttt tgatttttgg tgtgtgtgtc
     6001 ttgttgttag tatttgctag tcaaagtgat taaatagaat tcattcgcca ttcaggctgc
     6061 gcgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt
     6121 gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc
     6181 cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc
     6241 ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc
     6301 ggtgatgccg gccacgatgc gtccggcgta gaggatccac aggacgggtg tggtcgccat
     6361 gatcgcgtag tcgatagtgg ctccaagtag cgaagcgagc aggactgggc ggcggccaaa
     6421 gcggtcggac agtgctccga gaacgggtgc gcatagaaat tgcatcaacg catatagcgc
     6481 tagcagcacg ccatagtgac tggcgatgct gtcggaatgg acgatcacac tactcttctt
     6541 ttgcctatta taacatgaaa acgctttaac agaaagaata ggaaggtatc cgactgctgg
     6601 tattaaccct ctttctcaag ttatcatcgg caatgttgaa tggagtccat tcaaaacagc
     6661 atagctctaa aacatggaac tcaacaagtc tcagtgtgct gaagttttgg gaccattcaa
     6721 aacagcatag ctctaaaacg agacctttga gcttccgaga ctggtctcag ttttgggacc
     6781 attcaaaaca gcatagctct aaaacctcgt agactatttt tgtctaaaaa atttcgtaat
     6841 cgcactattt gtctcagcta gacttcagtc ttgaaaagcc cctgtattac tgcatttatt
     6901 aagagtatta taccatattt ttagttatta agaaataatc ttcatctaaa atatacttca
     6961 gtcacctcct agctgactca aatcaatgcg tgtttcataa agaccagtga tggattgatg
     7021 gataagagtg gcatctaaaa cttcttttgt agacgtatat cgtttacgat caattgttgt
     7081 atcaaaatat ttaaaagcag cgggagctcc aagattcgtc aacgtaaata aatgaataat
     7141 attttctgct tgttcacgta ttggtttgtc tctatgtttg ttatatgcac taagaacttt
     7201 atctaaattg gcatctgcta aaataacacg cttagaaaat tcactgattt gctcaataat
     7261 ctcatctaaa taatgcttat gctgctccac aaacaattgt ttttgttcgt tatcttctgg
     7321 actacccttc aacttttcat aatgactagc taaatataaa aaattcacat atttgcttgg
     7381 cagagccagc tcatttcctt tttgtaattc tccggcacta gccagcatcc gtttacgacc
     7441 gttttctaac tcaaaaagac tatatttagg tagtttaatg attaagtctt ttttaacttc
     7501 cttatatcct ttagcttcta aaaagtcaat cggatttttt tcaaaggaac ttctttccat
     7561 aattgtgatc cctagtaact ctttaacgga ttttaacttc ttcgatttcc ctttttccac
     7621 cttagcaacc actaggactg aataagctac cgttggacta tcaaaaccac catatttttt
     7681 tggatcccag tcttttttac gagcaataag cttgtccgaa tttctttttg gtaaaattga
     7741 ctccttggag aatccgcctg tctgtacttc tgttttcttg acaatattga cttggggcat
     7801 ggacaatact ttgcgcactg tggcaaaatc tcgcccttta tcccagacaa tttctccagt
     7861 ttccccatta gtttcgatta gagggcgttt gcgaatctct ccatttgcaa gtgtaatttc
     7921 tgttttgaag aagttcatga tattagagta aaagaaatat tttgcggttg ctttgcctat
     7981 ttcttgctca gacttagcaa tcattttacg aacatcataa actttataat caccatagac
     8041 aaactccgat tcaagttttg gatatttctt aatcaaagca gttccaacga cggcatttag
     8101 atacgcatca tgggcatgat ggtaattgtt aatctcacgt actttataga attggaaatc
     8161 ttttcggaag tcagaaacta atttagattt taaggtaatc actttaacct ctcgaataag
     8221 tttatcattt tcatcgtatt tagtattcat gcgactatcc aaaatttgtg ccacatgctt
     8281 agtgatttgg cgagtttcaa ccaattggcg tttgataaaa ccagctttat caagttcact
     8341 caaacctcca cgttcagctt tcgttaaatt atcaaactta cgttgagtga ttaacttggc
     8401 gtttagaagt tgtctccaat agtttttcat ctttttgact acttcttcac ttggaacgtt
     8461 atccgattta ccacgatttt tatcagaacg cgttaagacc ttattgtcta ttgaatcgtc
     8521 tttaaggaaa ctttgtggaa caatgtgatc gacatcataa tcacttaaac gattaatatc
     8581 taattcttgg tccacataca tgtctcttcc attttggaga taatagagat agagcttttc
     8641 attttgcaat tgagtatttt caacaggatg ctctttaaga atctgacttc ctaattcttt
     8701 gataccttct tcgattcgtt tcatacgctc tcgcgaattt ttctggccct tttgagttgt
     8761 ctgattttca cgtgccattt caataacgat attttctggc ttatgccgcc ccattacttt
     8821 gaccaattca tcaacaactt ttacagtctg taaaatacct tttttaatag cagggctacc
     8881 agctaaattt gcaatatgtt catgtaaact atcgccttgt ccagacactt gtgctttttg
     8941 aatgtcttct ttaaatgtca aactatcatc atggatcagc tgcataaaat tgcgattggc
     9001 aaaaccatct gatttcaaaa aatctaatat tgttttgcca gattgcttat ccctaatacc
     9061 attaatcaat tttcgagaca aacgtcccca accagtataa cggcgacgtt taagctgttt
     9121 catcacctta tcatcaaaga ggtgagcata tgttttaagt ctttcctcaa tcatctccct
     9181 atcttcaaat aaggtcaatg ttaaaacaat atcctctaag atatcttcat tttcttcatt
     9241 atccaaaaaa tctttatctt taataatttt tagcaaatca tggtaggtac ctaatgaagc
     9301 attaaatcta tcttcaactc ctgaaatttc aacactatca aaacattcta tttttttgaa
     9361 ataatcttct tttaattgct taacggttac ttttcgattt gttttgaaga gtaaatcaac
     9421 aatggctttc ttctgttcac ctgaaagaaa tgctggtttt cgcattcctt cagtaacata
     9481 tttgaccttt gtcaattcgt tataaaccgt aaaatactca taaagcaaac tatgttttgg
     9541 tagtactttt tcatttggaa gatttttatc aaagtttgtc atgcgttcaa taaatgattg
     9601 agctgaagca cctttatcga caacttcttc aaaattccat ggggtaattg tttcttcaga
     9661 cttccgagtc atccatgcaa aacgactatt gccacgcgcc aatggaccaa cataataagg
     9721 aattcgaaaa gtcaagattt tttcaatctt ctcacgattg tcttttaaaa atggataaaa
     9781 gtcttcttgt cttctcaaaa tagcatgcag ctcacccaag tgaatttgat ggggaataga
     9841 gccgttgtca aaggtccgtt gcttgcgcag caaatcttca cgatttagtt tcaccaataa
     9901 ttcctcagta ccatccattt tttctaaaat tggtttgata aatttataaa attcttcttg
     9961 gctagctccc ccatcaatat aacctgcata tccgtttttt gattgatcaa aaaagatttc
    10021 tttatacttt tctggaagtt gttgtcgaac taaagctttt aaaagagtca agtcttgatg
    10081 atgttcatcg tagcgtttaa tcattgaagc tgatagggga gccttagtta tttcagtatt
    10141 tactcttagg atatctgaaa gtaaaatagc atctgataaa ttcttagctg ccaaaaacaa
    10201 atcagcatat tgatctccaa tttgcgccaa taaattatct aaatcatcat cgtaagtatc
    10261 ttttgaaagc tgtaatttag catcttctgc caaatcaaaa tttgatttaa aattaggggt
    10321 caaacccaat gacaaagcaa tgagattccc aaataagcca tttttcttct caccggggag
    10381 ctgagcaatg agattttcta atcgtcttga tttactcaat cgtgcagaaa gaatcgcttt
    10441 agcatctact ccacttgcgt taatagggtt ttcttcaaat aattgattgt aggtttgtac
    10501 caactggata aatagtttgt ccacatcact attatcagga tttaaatctc cctcaatcaa
    10561 aaaatgacca cgaaacttaa tcatatgcgc taaggccaaa tagattaagc gcaaatccgc
    10621 tttatcagta gaatctacca atttttttcg cagatgatag atagttggat atttctcatg
    10681 ataagcaact tcatctacta tatttccaaa aataggatga cgttcatgct tcttgtcttc
    10741 ttccaccaaa aaagactctt caagtcgatg aaagaaacta tcatctactt tcgccatctc
    10801 atttgaaaaa atctcctgta gataacaaat acgattcttc cgacgtgtat accttctacg
    10861 agctgtccgt ttgagacgag tcgcttccgc tgtctctcca ctgtcaaata aaagagcccc
    10921 tataagattt tttttgatac tgtggcggtc tgtatttccc agaaccttga actttttaga
    10981 cggaacctta tattcatcag tgatcaccgc ccatccgacg ctatttgtgc cgatatctaa
    11041 gcctattgag tatttcttat ccatttttgc ctcctaaaat aaaaagttta aattaaatcc
    11101 ataatgagtt tgatgatttc aataatagtt ttaatgacct ccgaaattag tttaatatgc
    11161 tttaattttt ctttttcaaa atatctcttc aaaaaatatt acccaatact taataataaa
    11221 tagattataa cacaaaattc ttttaaaaag tagtttattt tgttatcatt ctatagtatt
    11281 aagtattgtt ttatggctga taaatttctt tgaatttctc cttgattatt tgttataaaa
    11341 gttataaaat aatcttgttg gaaccattca aaacagcata gcaagttaaa ataaggctag
    11401 tccgttatca acttgaaaaa gtggcaccga gtcggtgctt tttttgatac ttctattcta
    11461 ctctgactgc aaaccaaaaa aacaagcgct ttcaaaacgc ttgttttatc atttttaggg
    11521 aaattaatct cttaatcctt ttatcattct acatttaggc gctgccatct tgctaaacct
    11581 actaagctcc acaggatgat ttcgtaatcc cgcaagaggc ccggcagtac cggcataacc
    11641 aagcctatgc ctacagcatc cagggtgacg gtgccgagga tgacgatgag cgcattgtta
    11701 gatttcatac acggtgcctg actgcgttag caatttaact gtgataaact accgcattaa
    11761 agcttatcga tgataagctg tcaaacatga gaattacaac ttatatcgta tggggctgac
    11821 ttcaggtgct acatttgaag agataaattg cactgaaatc cagtcacgac gttgtaaaac
    11881 gacggccagt gaattcccgg ggatccgtcg acctgcagcc aagctttcgc gagctcgaga
    11941 tctagatatc gatgaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc
    12001 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
    12061 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
    12121 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc g
//
