LOCUS       sequence-202234-       14173 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Clostridium difficile CRISPR-cas9 mutagenesis plasmid traJ oriT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14173)
  AUTHORS   McAllister KN, Bouillaut L, Kahn JN, Self WT, Sorg JA
  TITLE     Using CRISPR-Cas9-mediated genome editing to generate C. difficile 
            mutants defective in selenoproteins synthesis.
  JOURNAL   Sci Rep. 2017 Nov 7;7(1):14672. doi: 10.1038/s41598-017-15236-5.
  PUBMED    29116155
REFERENCE   2  (bases 1 to 14173)
  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: "Sci Rep."; 
            date: "2017-11-7"; volume: "7(1):14672. doi"; pages: " 
            10.1038/s41598-017-15236-5"
COMMENT     Sequence Label: pJK02
FEATURES             Location/Qualifiers
     source          1..14173
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     63..79
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     63..79
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(2117..2740)
                     /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     2645..2664
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     protein_bind    2847..2865
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     CDS             2938..7044
                     /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             2938..7041
                     /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"
     primer_bind     complement(7054..7071)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7054..7070)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7063..7085)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_RNA        12344..12419
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      12617..13205
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     13106..13125
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     13359..13376
                     /label=L4440
                     /note="L4440 vector, forward primer"
     oriT            13526..13635
                     /note="incP origin of transfer"
     CDS             13668..14039
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
ORIGIN
        1 cctgcaggat aaaaaaattg tagataaatt ttataaaata gttttatcta caattttttt
       61 atcaggaaac agctatgacc gcggccgcga cgtgattttt aatgggtaat ttaagtgtga
      121 aatttggaag tgtagaattt aaaaatccag taataatggc gtctggaact tttggatttg
      181 gaaaagaata taatgagatt tatgatatac aaaaacttgg tggtataagt agcaagggtc
      241 ttactttaaa taaaaaacct ggtaacaatg gtatgagagt tcatgagaca ccatccggta
      301 tgatgaatag tgttgggctt gaaaaccctg gagttcaagg atttatagac tatgaattac
      361 ctttttttag taaactagac ttagtcagaa tagctaatgt tggaggtggg acattagaag
      421 actacttatt aggggttcaa atgttaaatg acaagcctat agacattatt gaattaaata
      481 tatcttgccc aaatgtcaaa gctggtggaa tggcttttgg tataaaaaat gaagtggcaa
      541 gagaagttgt tcgtgaggta agaaacataa cgaaactgcc tttagtaata aaattatcac
      601 ccaatgcaga agatattgta ggaatggcaa aagtttgtga agaagaaggt gctgatggag
      661 tttcattagt taatactttt aaagcaatgg caattgatat aaaaaataga agaccagtat
      721 ttgaaaatgt atatgctgga ttatctggac cagctataaa acctatagct ttgagaatgg
      781 tacatgaagt gtgtaaaaat gtaaatatac ctgtaatggg aatgggcgga ataactaaag
      841 ccacagatgc tatagagttt ataatggcag gagctacttg tatccaagtt ggaactgcaa
      901 attttatgaa tcctagaata ggtattgaaa taatagatgg gattaatgaa tttatggata
      961 gagaaggaat aaaaagttta gacgaaataa gaggaattat ataaggtaga ataaatagta
     1021 ttttaaagat aaaaaatata gattaaataa acaataattt tataacatta acatggagga
     1081 aaaataatta aaaacttaat tatttatagt gttacttaaa aaatgtaaat tttttagagt
     1141 aaaaatttgt aatataacat ataacttaaa aataaaaagc agttagcctt ataaatgaag
     1201 gttagctgtt ttttttgtta aaaatatatt tacaaaataa gagatagtag cagtacaata
     1261 taagtataag gtaaaagtat tatatattag ggaggggtaa gtatgaaaaa tcaattaaat
     1321 gaaaatcata aaaaggttgt attttttgga gtaggtgctg taggagctac ttttgcagaa
     1381 caatttttta actctaaata tgattttaaa attctttgtg ataatgaaag aaagaaaaga
     1441 tatttagaag aaggatttat aataaatgga aaaaggtatg attttaatta tgtaactaaa
     1501 gatgagtata aacaagaggc tgattttata attataggtc taaaatataa taatttaaaa
     1561 gaaaatataa aagaattaga tggattagtt ggaaaaaata cagttataat gtctctgcta
     1621 aatggagttg atagcgaaga gataatagga gaaagatttg gaattggaaa aatggtatat
     1681 tcatatgtta ccaatataga tgcaaagaaa atcaataata atattataca cactactaat
     1741 gggataattg tatttggtaa caaagataat agtgaagata gaaaaactaa tataataact
     1801 gaagtctttg atgatgtaaa tatagaatat attttatcaa aagatattca acgagatatg
     1861 tggtggaagt acatggttaa tattggtgta aatcaaactt cagctatact tggtgcacct
     1921 tatggagttt ttcagagttc tgagcattta agagaattag caaaatctgc aatgagggaa
     1981 gttgttgcta tagcacaagc aaaagacata tctcttacag aagatgatgt tgaaaattca
     2041 ttacatagaa tactagaaca ttcaaaagaa ggaagaacat caatgcttct cgaggcctgc
     2101 agacatgcgc gatcgcagac ccactttcac atttaagttg tttttctaat ccgcatatga
     2161 tcaattcaag gccgaataag aaggctggct ctgcaccttg gtgatcaaat aattcgatag
     2221 cttgtcgtaa taatggcggc atactatcag tagtaggtgt ttccctttct tctttagcga
     2281 cttgatgctc ttgatcttcc aatacgcaac ctaaagtaaa atgccccaca gcgctgagtg
     2341 catataatgc attctctagt gaaaaacctt gttggcataa aaaggctaat tgattttcga
     2401 gagtttcata ctgtttttct gtaggccgtg tacctaaatg tacttttgct ccatcgcgat
     2461 gacttagtaa agcacatcta aaacttttag cgttattacg taaaaaatct tgccagcttt
     2521 ccccttctaa agggcaaaag tgagtatggt gcctatctaa catctcaatg gctaaggcgt
     2581 cgagcaaagc ccgcttattt tttacatgcc aatacaatgt aggctgctct acacctagct
     2641 tctgggcgag tttacgggtt gttaaacctt cgattccgac ctcattaagc agctctaatg
     2701 cgctgttaat cactttactt ttatctaatc tagacatcat taattcctcc tttttgttga
     2761 cattatatca ttgatagagt tatttgtcaa actagttttt tatttcgatg ccctggactt
     2821 catgaaaaac taaaaaaaat attgacactc tatcattgat agagtataat taaaataagc
     2881 ttgatcgtag cgttaacaga tctgagctca ataatactag gaggtttttt taattaaatg
     2941 gataaaaaat atagtatagg attagatata ggaacaaata gtgtaggatg ggcagtaata
     3001 actgatgaat ataaagtacc aagtaaaaaa tttaaagtat taggaaatac agatagacat
     3061 agtataaaaa aaaatttaat aggtgcatta ctttttgata gtggtgaaac agcagaagca
     3121 acaagactta aaagaacagc aagaagaaga tatactagaa gaaaaaatag aatatgttat
     3181 ttacaagaaa tatttagtaa tgaaatggca aaagtagatg atagtttttt tcatagatta
     3241 gaagaaagtt ttttagtaga agaagataaa aaacacgaaa gacatccaat atttggaaat
     3301 atagtagatg aagtagcata tcatgaaaaa tatccaacaa tatatcattt aagaaaaaaa
     3361 cttgtagata gtacagataa agcagattta agacttatat atcttgcatt agcacatatg
     3421 ataaaattta gaggacattt tcttatagaa ggtgatttaa atccagataa tagtgatgta
     3481 gataaacttt ttatacaatt agtacaaaca tataatcaat tatttgaaga aaatccaata
     3541 aatgcaagtg gtgtagatgc aaaagcaata ttaagtgcaa gattaagtaa aagtagaaga
     3601 ttagaaaatc ttatagcaca attacctgga gaaaaaaaaa atggattatt tggtaatctt
     3661 atagctttaa gtttaggatt aacaccaaat tttaaatcaa attttgatct tgcagaagat
     3721 gcaaaacttc aattaagtaa agatacatat gatgatgatt tagataattt acttgcacaa
     3781 ataggtgatc aatatgcaga tttatttctt gcagcaaaaa atctttctga tgcaatactt
     3841 ttaagtgata tacttagagt aaatacagaa ataacaaaag caccattatc tgcaagtatg
     3901 ataaaaagat atgatgaaca tcatcaagat ttaacattac ttaaagcatt agtaagacaa
     3961 caattaccag aaaaatataa agaaattttt tttgatcaaa gtaaaaatgg atatgcagga
     4021 tatatagatg gtggtgcaag tcaagaagaa ttttataaat ttattaaacc aatattagaa
     4081 aaaatggatg gaacagaaga attattagta aaacttaata gagaagattt acttagaaaa
     4141 caaagaacat ttgataatgg aagtatacca catcaaatac atttaggtga attacatgca
     4201 atattaagaa gacaagaaga tttttatcca tttcttaaag ataatagaga aaaaattgaa
     4261 aaaatactta catttagaat accatattat gtaggaccat tagcaagagg aaatagtaga
     4321 tttgcttgga tgacaagaaa aagtgaagaa acaataacac catggaattt tgaagaagta
     4381 gtagataaag gagcatctgc acaaagtttt atagaaagaa tgacaaattt tgataaaaat
     4441 cttccaaatg aaaaagtatt accaaaacat agtttattat atgaatattt tacagtatat
     4501 aatgaattaa caaaagtaaa atatgtaaca gaaggtatga gaaaaccagc atttttaagt
     4561 ggtgaacaaa aaaaagctat agtagattta ttatttaaaa caaatagaaa agtaacagta
     4621 aaacaattaa aagaagatta ttttaaaaaa attgaatgtt ttgatagtgt agaaatatct
     4681 ggtgtagaag atagatttaa tgcaagttta ggaacatatc atgatttact taaaataata
     4741 aaggataaag attttttaga taatgaagaa aatgaagata ttttagaaga tatagtatta
     4801 acattaactt tatttgaaga tagagaaatg atagaagaaa gattaaaaac ttatgctcat
     4861 ttatttgatg ataaagtaat gaaacaactt aaaagaagaa gatatacagg ttggggaaga
     4921 ttaagtagaa aacttataaa tggaataaga gataaacaaa gtggaaaaac aatattagat
     4981 tttcttaaaa gtgatggatt tgcaaataga aattttatgc aattaataca tgatgatagt
     5041 ttaacattta aagaagatat tcaaaaagca caagtaagtg gacaaggtga tagtttacat
     5101 gaacatatag caaatttagc aggatctcca gcaataaaaa aaggtatatt acaaacagta
     5161 aaagtagttg atgaattagt aaaagtaatg ggtagacata aaccagaaaa tatagttata
     5221 gaaatggcaa gagaaaatca aacaacacaa aaaggacaaa aaaatagtag agaaagaatg
     5281 aaaagaatag aagaaggtat aaaagaatta ggaagtcaaa tacttaaaga acatccagta
     5341 gaaaatacac aacttcaaaa tgaaaaactt tatttatatt atttacaaaa tggaagagat
     5401 atgtatgtag atcaagaatt agatataaat agattaagtg attatgatgt agatcatata
     5461 gtaccacaaa gttttcttaa agatgattct atagataata aagtattaac aagatctgat
     5521 aaaaatagag gtaaaagtga taatgtacca agtgaagaag ttgtaaaaaa aatgaaaaat
     5581 tattggagac aattacttaa tgcaaaactt ataacacaaa gaaaatttga taatcttaca
     5641 aaagcagaaa gaggtggatt atctgaatta gataaagctg gatttataaa aagacaatta
     5701 gtagaaacta gacaaataac aaaacatgta gcacaaatat tagatagtag aatgaataca
     5761 aaatatgatg aaaatgataa gcttataaga gaagtaaaag ttataacatt aaaaagtaaa
     5821 ttagtaagtg attttagaaa agattttcaa ttttataaag taagagaaat aaataattat
     5881 catcatgcac atgatgcata tcttaatgca gtagtaggaa cagcattaat aaaaaaatat
     5941 cctaaattag aatctgaatt tgtatatgga gattataaag tatatgatgt aagaaaaatg
     6001 atagcaaaaa gtgaacaaga aataggaaaa gcaacagcaa agtacttttt ttattctaat
     6061 ataatgaatt tttttaagac agaaataact ttagcaaatg gtgaaataag aaaaagacct
     6121 ttaatagaaa ctaatggtga aacaggtgaa atagtatggg ataaaggtag agattttgca
     6181 acagtaagaa aagtattaag tatgccacaa gtaaatatag taaaaaaaac agaagtacaa
     6241 actggaggat ttagtaaaga aagtatactt ccaaaaagaa attcagataa acttattgca
     6301 agaaaaaaag attgggatcc aaaaaaatat ggtggatttg atagtccaac agtagcatat
     6361 agtgtattag tagtagcaaa agtagaaaaa ggtaaatcta aaaaacttaa aagtgtaaaa
     6421 gaattacttg gaataacaat aatggaaaga agtagttttg aaaaaaatcc tattgatttt
     6481 ttagaagcaa aaggatataa agaagttaaa aaagatttaa taataaaact tccaaaatat
     6541 tctttatttg aattagaaaa tggaagaaaa agaatgttag caagtgctgg tgaattacaa
     6601 aaaggtaatg aattagcatt accatctaaa tatgtaaatt ttctttattt agcaagtcat
     6661 tatgaaaaac ttaaaggatc tcctgaagat aatgaacaaa aacaattatt tgtagaacaa
     6721 cataaacatt atttagatga aataattgaa caaataagtg aattttctaa aagagtaata
     6781 ttagcagacg caaatttaga taaagtatta tctgcttata ataaacatag agataaacca
     6841 ataagagaac aagcagaaaa tataatacat ctttttacat taacaaattt aggtgcacca
     6901 gcagcattta aatattttga tacaacaata gatagaaaaa gatatacaag tacaaaagaa
     6961 gtattagacg caacattaat acatcaaagt ataacaggat tatatgaaac aagaatagat
     7021 ttatctcaat taggtggtga ttaaaagctt ggcactggcc gtcgttttac aacgtcgtga
     7081 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     7141 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     7201 tggcgaatgg cgctagcata aaaataagaa gcctgcattt gcaggcttct tatttttatg
     7261 gcgcgcccgc ccttaagtct aaaaattagg ggagatgtaa ggatttggga aaaatagaag
     7321 atgttataat cataaatatg gtattcgtag gcttaaagtc aaaaaggagg tgaaatataa
     7381 atagattttt agctaaatta agtaagaaat aggaggagat ttattgaaca aaaaattaga
     7441 aaaaccattt gtatataaga gagagtacga tttgactgga tatgatgttg aaattttaca
     7501 aaaatatgag ttagaacaag caatatatgt ttatgttggg agtagttgtg catataacat
     7561 gagagctaga agtagtaaat ggagatacca tataagaaca aataataagt ctatatgttg
     7621 taacattaaa aattttatac ataacttgga attgttttat aaaatggaat taaagttgtc
     7681 agataatatt attaatgata agctatacta tagcaatata gcagagtttg aagaatttga
     7741 aacactagaa aaagctagag aggtagaaag tactattata agtcaatatc aatttttaga
     7801 ttctataaat cacatgttaa aacaaaaaat aattttattg agtaataagg atagtgtgtt
     7861 aaacataact aaaaatggaa atacaaatta tttgaaagta aaaaataaat acatagaaaa
     7921 acataagaac aagccaataa tgagatacca tatcaactgt caattcaata cagatggaag
     7981 tgtcaaaagt attacacagg agtttgaacc aatattggaa ttaaacaaaa aaaataccct
     8041 aagccgacca agcagagtat ttttaaaata atattttaag ataacaacaa aatgagataa
     8101 tactactaga caatgacaac tcaactacca attgagttta tggagctacc aactccaata
     8161 tcggtctaac tgattaagta tctgtagtta tataataata ttgctatcaa ttttagcatc
     8221 ttaacaatat tattatacat actaagctaa aattattcaa tagttgtaaa agttgattag
     8281 tcaataagta tatatttaat gtagtgttat ctcttaaaaa aactagataa ggagataata
     8341 aatatatgga acaattagat tcaaaatata agttgaaaaa atttctaatg gcagtattta
     8401 gagatggtat aggacaagga aataatctta ttgataatga atatgttaga gtatttcaaa
     8461 ataataaaag taatagtaaa caattagaac tcggagaaga atttaaagaa tatagtaaaa
     8521 caactttttt taaaaatata gatgatatag tagaatttac cttcgcaaaa aatatttatt
     8581 atgaaaatac attttttaac ctatgtacta ctgatggaaa agcaggaacc aatgaaaact
     8641 taataaatag atatgcatta ggatttgatt ttgacaaaaa agaattagga caaggtttta
     8701 attataaaga tataattaat ttatttacta agataggatt acattatcat atcctagttg
     8761 atagtggaaa tggattccat gtttatgtgc taattaataa aactaataac attaagttag
     8821 tatcagaagt tacaaataca ttaataaata aattgggtgc agataaacaa gcaaatttat
     8881 ctactcaagt attaagagta ccttatacat ataatattaa aaatactact aaacaagtaa
     8941 aaataataca ccaagacaaa aatatatata gatatgacat agaaaagtta gctaaaaaat
     9001 attgcaaaga tgtaaaaaca gtaggtaata ctaatacaaa atatatatta gatagtaagc
     9061 taccaaattg tatagtagat attttaaaaa atggtagtaa agatggacat aaaaacctag
     9121 atttgcaaaa aatagttgtg actttaagat tgaggaataa aagtttaagt caagtaatat
     9181 ccgttgctag agaatggaac tatatatcac aaaatagtct ttcaaatagt gagctagaat
     9241 atcaagtcaa gtatatgtat gagaaactta aaacggttaa ttttggttgt actggttgtg
     9301 agtttaatag tgattgttgg aataaaatag aatcagattt tatatatagt gatgaagata
     9361 ctttgttcaa tatgccacat aagcactcaa aggatttgaa atataagaat aggaaagggg
     9421 ttaaaataat gactggtaat caattgttta tctataatgt gttacttaac aataaagata
     9481 gagaattaaa catagacgat ataatggagc tgataaccta taaacgtaag aagaaagtta
     9541 aaaacattgt tatgagtgaa aagacattaa gagaaacatt aaaagaactt caacataatg
     9601 attatattac aaaaacaaaa ggtgttacaa agctaggaat aaaagataca tacaatgtaa
     9661 aagaagttag atgtaatata gataaacaat atactattag ttactttgtt accatggcag
     9721 taatttgggg aataatttca actgaagaat taagattata tactcacatg agatataagc
     9781 aagatttatt ggtcaaagat gataaaataa aaggaaatat attaagaatt aatcaagagg
     9841 aattagcaaa agatttagga gtaacacagc aaagaatttc aaatatgata gaatctttat
     9901 tagatactaa aattttagat gtatgggaaa ctaaaataaa tgatagagga tttatgtact
     9961 atacatatag attaaacaag tagatttttg ataggattag aattgatttt ctagtcctat
    10021 ttttatgcaa aaaaactaat taataaaaat ttcttttggt aaaataattg tacgagaatt
    10081 gcaaaaaaaa aatggcatca aagtattgaa attaagccgt tttaaaaatt tcttttggta
    10141 aaataattct acatatatat gtagtatata tatatatgtt ttttagagaa tgtataacta
    10201 gaatatagag ctagaatata gagaatgtat aactagaata tagagctaga atatagagaa
    10261 tgtataacta gaatatagag ctagaatata gagaatgtat aactagaata tagagctaga
    10321 atatagagaa tgtataacta gaatatagag ctagaatata gagaatgtat aactagaata
    10381 tagagctaga atatagagaa tgtataacta gaatatagag ctagaatcct aattagtagg
    10441 tgctttttta aaacaagtta aaaatcaaaa atagtattag taagcattgg aaatgctaga
    10501 ttctaaaata gaaaagtaaa aaattggtgc actatctaaa cttatctata tcgctttttc
    10561 cgtcgtttgg ttctctagtt acgatacagg ggatatgctt atattgagtt atagtactaa
    10621 tcagtgctta atatagttaa taaaattata gttaccatag tttagtaact atgatgtatg
    10681 ttagttagaa acttgcattt cggccggcca gtgggcaagt tgaaaaattc acaaaaatgt
    10741 ggtataatat ctttgttcat tagagcgata aacttgaatt tgagagggaa cttagatggt
    10801 atttgaaaaa attgataaaa atagttggaa cagaaaagag tattttgacc actactttgc
    10861 aagtgtacct tgtacctaca gcatgaccgt taaagtggat atcacacaaa taaaggaaaa
    10921 gggaatgaaa ctatatcctg caatgcttta ttatattgca atgattgtaa accgccattc
    10981 agagtttagg acggcaatca atcaagatgg tgaattgggg atatatgatg agatgatacc
    11041 aagctataca atatttcaca atgatactga aacattttcc agcctttgga ctgagtgtaa
    11101 gtctgacttt aaatcatttt tagcagatta tgaaagtgat acgcaacggt atggaaacaa
    11161 tcatagaatg gaaggaaagc caaatgctcc ggaaaacatt tttaatgtat ctatgatacc
    11221 gtggtcaacc ttcgatggct ttaatctgaa tttgcagaaa ggatatgatt atttgattcc
    11281 tatttttact atggggaaat attataaaga agataacaaa attatacttc ctttggcaat
    11341 tcaagttcat cacgcagtat gtgacggatt tcacatttgc cgttttgtaa acgaattgca
    11401 ggaattgata aatagttaac ttcaggtttg tctgtaacta aaaacaagta tttaagcaaa
    11461 aacatcgtag aaatacggtg ttttttgtta ccctaagttt cataaaaata agaagcctgc
    11521 aaatgcaggc ttcttatttt tatggtttaa acggttttag ctgggatatc ggcaaaacaa
    11581 gcaaatacta ctatagaagc agttaaaagg gctatttttt ataatctgga gcatagacaa
    11641 gaagttagag aagcaattaa aaaagaaaat ataagaaaaa ttcttattat tgggacatca
    11701 aaaaaaatgg ttaatcaaat tgtagagaga ttgtcaattg gaagggtctg taagtttata
    11761 aatatacaag atataagtac taaatatgaa atagaaatag caaaacaagc tagaaaagaa
    11821 ggaaatcatg taattcctgt tccagccgtt gagataaaaa gtatggcaag tggtctcagt
    11881 ataaattctt tgaagagact ttttagaaaa ggaaataata ggaatatggt agttttagaa
    11941 aaaacaatta taagaccaac ttttagttat ataggaaagt tttatattag tgctgatgta
    12001 attaaacaga ttattgaata tgaagtatac aactttggaa gcattgatag aataaataaa
    12061 atcaatatag agaattgcaa taattttatg aatatttttg taagtataaa tatcaatgat
    12121 ttagaaacta taaaaaaaat tgaagacata caacagacaa taaaaaagtc tattgagaaa
    12181 atgactttag ttaatgtaca aaaaatagat attaacattc ataaaattaa aaaaataaac
    12241 ttcctccaat gttaaaaaat aaactgagaa aatgatatac taatttgcaa taaagtgtgc
    12301 tataattaaa ctgtaaaggt accgaaaagt gatgcattgt tgggttttag agctagaaat
    12361 agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct
    12421 ttttttctat ggagaaatct agatcagcat gatgtctgac tagacgcgta agctctgcaa
    12481 ctatttttag atggttgcag agcttatttt tattttttat taaactcctt tttgataatc
    12541 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
    12601 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
    12661 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
    12721 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
    12781 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
    12841 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
    12901 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
    12961 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
    13021 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
    13081 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
    13141 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
    13201 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
    13261 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
    13321 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
    13381 cggaagagcg cccaatacgc agggccccct gcttcggggt cattatagcg attttttcgg
    13441 tatatccatc ctttttcgca cgatatacag gattttgcca aagggttcgt gtagactttc
    13501 cttggtgtat ccaacggcgt cagccgggca ggataggtga agtaggccca cccgcgagcg
    13561 ggtgttcctt cttcactgtc ccttattcgc acctggcggt gctcaacggg aatcctgctc
    13621 tgcgaggctg gccggctacc gccggcgtaa cagatgaggg caagcggatg gctgatgaaa
    13681 ccaagccaac caggaagggc agcccaccta tcaaggtgta ctgccttcca gacgaacgaa
    13741 gagcgattga ggaaaaggcg gcggcggccg gcatgagcct gtcggcctac ctgctggccg
    13801 tcggccaggg ctacaaaatc acgggcgtcg tggactatga gcacgtccgc gagctggccc
    13861 gcatcaatgg cgacctgggc cgcctgggcg gcctgctgaa actctggctc accgacgacc
    13921 cgcgcacggc gcggttcggt gatgccacga tcctcgccct gctggcgaag atcgaagaga
    13981 agcaggacga gcttggcaag gtcatgatgg gcgtggtccg cccgagggca gagccatgac
    14041 ttttttagcc gctaaaacgg ccggggggtg cgcgtgattg ccaagcacgt ccccatgcgc
    14101 tccatcaaga agagcgactt cgcggagctg gtgaagtaca tcaccgacga gcaaggcaag
    14161 accgatcggg ccc
//
