LOCUS       sequence-202268-       13517 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Clostridium difficile CRISPR-cas9 mutagenesis plasmid Tn916 oriT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 13517)
  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 13517)
  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: pKM126
FEATURES             Location/Qualifiers
     source          1..13517
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     109..125
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     109..125
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(2163..2786)
                     /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     2691..2710
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     protein_bind    2893..2911
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     CDS             2984..7090
                     /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             2984..7087
                     /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(7100..7117)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7100..7116)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7109..7131)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_RNA        12390..12465
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      12663..13251
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     13152..13171
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     13405..13422
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ttctgcagat tacctaataa tttatctaca ttccctttag gggccccctg caggataaaa
       61 aaattgtaga taaattttat aaaatagttt tatctacaat ttttttatca ggaaacagct
      121 atgaccgcgg ccgcgacgtg atttttaatg ggtaatttaa gtgtgaaatt tggaagtgta
      181 gaatttaaaa atccagtaat aatggcgtct ggaacttttg gatttggaaa agaatataat
      241 gagatttatg atatacaaaa acttggtggt ataagtagca agggtcttac tttaaataaa
      301 aaacctggta acaatggtat gagagttcat gagacaccat ccggtatgat gaatagtgtt
      361 gggcttgaaa accctggagt tcaaggattt atagactatg aattaccttt ttttagtaaa
      421 ctagacttag tcagaatagc taatgttgga ggtgggacat tagaagacta cttattaggg
      481 gttcaaatgt taaatgacaa gcctatagac attattgaat taaatatatc ttgcccaaat
      541 gtcaaagctg gtggaatggc ttttggtata aaaaatgaag tggcaagaga agttgttcgt
      601 gaggtaagaa acataacgaa actgccttta gtaataaaat tatcacccaa tgcagaagat
      661 attgtaggaa tggcaaaagt ttgtgaagaa gaaggtgctg atggagtttc attagttaat
      721 acttttaaag caatggcaat tgatataaaa aatagaagac cagtatttga aaatgtatat
      781 gctggattat ctggaccagc tataaaacct atagctttga gaatggtaca tgaagtgtgt
      841 aaaaatgtaa atatacctgt aatgggaatg ggcggaataa ctaaagccac agatgctata
      901 gagtttataa tggcaggagc tacttgtatc caagttggaa ctgcaaattt tatgaatcct
      961 agaataggta ttgaaataat agatgggatt aatgaattta tggatagaga aggaataaaa
     1021 agtttagacg aaataagagg aattatataa ggtagaataa atagtatttt aaagataaaa
     1081 aatatagatt aaataaacaa taattttata acattaacat ggaggaaaaa taattaaaaa
     1141 cttaattatt tatagtgtta cttaaaaaat gtaaattttt tagagtaaaa atttgtaata
     1201 taacatataa cttaaaaata aaaagcagtt agccttataa atgaaggtta gctgtttttt
     1261 ttgttaaaaa tatatttaca aaataagaga tagtagcagt acaatataag tataaggtaa
     1321 aagtattata tattagggag gggtaagtat gaaaaatcaa ttaaatgaaa atcataaaaa
     1381 ggttgtattt tttggagtag gtgctgtagg agctactttt gcagaacaat tttttaactc
     1441 taaatatgat tttaaaattc tttgtgataa tgaaagaaag aaaagatatt tagaagaagg
     1501 atttataata aatggaaaaa ggtatgattt taattatgta actaaagatg agtataaaca
     1561 agaggctgat tttataatta taggtctaaa atataataat ttaaaagaaa atataaaaga
     1621 attagatgga ttagttggaa aaaatacagt tataatgtct ctgctaaatg gagttgatag
     1681 cgaagagata ataggagaaa gatttggaat tggaaaaatg gtatattcat atgttaccaa
     1741 tatagatgca aagaaaatca ataataatat tatacacact actaatggga taattgtatt
     1801 tggtaacaaa gataatagtg aagatagaaa aactaatata ataactgaag tctttgatga
     1861 tgtaaatata gaatatattt tatcaaaaga tattcaacga gatatgtggt ggaagtacat
     1921 ggttaatatt ggtgtaaatc aaacttcagc tatacttggt gcaccttatg gagtttttca
     1981 gagttctgag catttaagag aattagcaaa atctgcaatg agggaagttg ttgctatagc
     2041 acaagcaaaa gacatatctc ttacagaaga tgatgttgaa aattcattac atagaatact
     2101 agaacattca aaagaaggaa gaacatcaat gcttctcgag gcctgcagac atgcgcgatc
     2161 gcagacccac tttcacattt aagttgtttt tctaatccgc atatgatcaa ttcaaggccg
     2221 aataagaagg ctggctctgc accttggtga tcaaataatt cgatagcttg tcgtaataat
     2281 ggcggcatac tatcagtagt aggtgtttcc ctttcttctt tagcgacttg atgctcttga
     2341 tcttccaata cgcaacctaa agtaaaatgc cccacagcgc tgagtgcata taatgcattc
     2401 tctagtgaaa aaccttgttg gcataaaaag gctaattgat tttcgagagt ttcatactgt
     2461 ttttctgtag gccgtgtacc taaatgtact tttgctccat cgcgatgact tagtaaagca
     2521 catctaaaac ttttagcgtt attacgtaaa aaatcttgcc agctttcccc ttctaaaggg
     2581 caaaagtgag tatggtgcct atctaacatc tcaatggcta aggcgtcgag caaagcccgc
     2641 ttatttttta catgccaata caatgtaggc tgctctacac ctagcttctg ggcgagttta
     2701 cgggttgtta aaccttcgat tccgacctca ttaagcagct ctaatgcgct gttaatcact
     2761 ttacttttat ctaatctaga catcattaat tcctcctttt tgttgacatt atatcattga
     2821 tagagttatt tgtcaaacta gttttttatt tcgatgccct ggacttcatg aaaaactaaa
     2881 aaaaatattg acactctatc attgatagag tataattaaa ataagcttga tcgtagcgtt
     2941 aacagatctg agctcaataa tactaggagg tttttttaat taaatggata aaaaatatag
     3001 tataggatta gatataggaa caaatagtgt aggatgggca gtaataactg atgaatataa
     3061 agtaccaagt aaaaaattta aagtattagg aaatacagat agacatagta taaaaaaaaa
     3121 tttaataggt gcattacttt ttgatagtgg tgaaacagca gaagcaacaa gacttaaaag
     3181 aacagcaaga agaagatata ctagaagaaa aaatagaata tgttatttac aagaaatatt
     3241 tagtaatgaa atggcaaaag tagatgatag tttttttcat agattagaag aaagtttttt
     3301 agtagaagaa gataaaaaac acgaaagaca tccaatattt ggaaatatag tagatgaagt
     3361 agcatatcat gaaaaatatc caacaatata tcatttaaga aaaaaacttg tagatagtac
     3421 agataaagca gatttaagac ttatatatct tgcattagca catatgataa aatttagagg
     3481 acattttctt atagaaggtg atttaaatcc agataatagt gatgtagata aactttttat
     3541 acaattagta caaacatata atcaattatt tgaagaaaat ccaataaatg caagtggtgt
     3601 agatgcaaaa gcaatattaa gtgcaagatt aagtaaaagt agaagattag aaaatcttat
     3661 agcacaatta cctggagaaa aaaaaaatgg attatttggt aatcttatag ctttaagttt
     3721 aggattaaca ccaaatttta aatcaaattt tgatcttgca gaagatgcaa aacttcaatt
     3781 aagtaaagat acatatgatg atgatttaga taatttactt gcacaaatag gtgatcaata
     3841 tgcagattta tttcttgcag caaaaaatct ttctgatgca atacttttaa gtgatatact
     3901 tagagtaaat acagaaataa caaaagcacc attatctgca agtatgataa aaagatatga
     3961 tgaacatcat caagatttaa cattacttaa agcattagta agacaacaat taccagaaaa
     4021 atataaagaa attttttttg atcaaagtaa aaatggatat gcaggatata tagatggtgg
     4081 tgcaagtcaa gaagaatttt ataaatttat taaaccaata ttagaaaaaa tggatggaac
     4141 agaagaatta ttagtaaaac ttaatagaga agatttactt agaaaacaaa gaacatttga
     4201 taatggaagt ataccacatc aaatacattt aggtgaatta catgcaatat taagaagaca
     4261 agaagatttt tatccatttc ttaaagataa tagagaaaaa attgaaaaaa tacttacatt
     4321 tagaatacca tattatgtag gaccattagc aagaggaaat agtagatttg cttggatgac
     4381 aagaaaaagt gaagaaacaa taacaccatg gaattttgaa gaagtagtag ataaaggagc
     4441 atctgcacaa agttttatag aaagaatgac aaattttgat aaaaatcttc caaatgaaaa
     4501 agtattacca aaacatagtt tattatatga atattttaca gtatataatg aattaacaaa
     4561 agtaaaatat gtaacagaag gtatgagaaa accagcattt ttaagtggtg aacaaaaaaa
     4621 agctatagta gatttattat ttaaaacaaa tagaaaagta acagtaaaac aattaaaaga
     4681 agattatttt aaaaaaattg aatgttttga tagtgtagaa atatctggtg tagaagatag
     4741 atttaatgca agtttaggaa catatcatga tttacttaaa ataataaagg ataaagattt
     4801 tttagataat gaagaaaatg aagatatttt agaagatata gtattaacat taactttatt
     4861 tgaagataga gaaatgatag aagaaagatt aaaaacttat gctcatttat ttgatgataa
     4921 agtaatgaaa caacttaaaa gaagaagata tacaggttgg ggaagattaa gtagaaaact
     4981 tataaatgga ataagagata aacaaagtgg aaaaacaata ttagattttc ttaaaagtga
     5041 tggatttgca aatagaaatt ttatgcaatt aatacatgat gatagtttaa catttaaaga
     5101 agatattcaa aaagcacaag taagtggaca aggtgatagt ttacatgaac atatagcaaa
     5161 tttagcagga tctccagcaa taaaaaaagg tatattacaa acagtaaaag tagttgatga
     5221 attagtaaaa gtaatgggta gacataaacc agaaaatata gttatagaaa tggcaagaga
     5281 aaatcaaaca acacaaaaag gacaaaaaaa tagtagagaa agaatgaaaa gaatagaaga
     5341 aggtataaaa gaattaggaa gtcaaatact taaagaacat ccagtagaaa atacacaact
     5401 tcaaaatgaa aaactttatt tatattattt acaaaatgga agagatatgt atgtagatca
     5461 agaattagat ataaatagat taagtgatta tgatgtagat catatagtac cacaaagttt
     5521 tcttaaagat gattctatag ataataaagt attaacaaga tctgataaaa atagaggtaa
     5581 aagtgataat gtaccaagtg aagaagttgt aaaaaaaatg aaaaattatt ggagacaatt
     5641 acttaatgca aaacttataa cacaaagaaa atttgataat cttacaaaag cagaaagagg
     5701 tggattatct gaattagata aagctggatt tataaaaaga caattagtag aaactagaca
     5761 aataacaaaa catgtagcac aaatattaga tagtagaatg aatacaaaat atgatgaaaa
     5821 tgataagctt ataagagaag taaaagttat aacattaaaa agtaaattag taagtgattt
     5881 tagaaaagat tttcaatttt ataaagtaag agaaataaat aattatcatc atgcacatga
     5941 tgcatatctt aatgcagtag taggaacagc attaataaaa aaatatccta aattagaatc
     6001 tgaatttgta tatggagatt ataaagtata tgatgtaaga aaaatgatag caaaaagtga
     6061 acaagaaata ggaaaagcaa cagcaaagta ctttttttat tctaatataa tgaatttttt
     6121 taagacagaa ataactttag caaatggtga aataagaaaa agacctttaa tagaaactaa
     6181 tggtgaaaca ggtgaaatag tatgggataa aggtagagat tttgcaacag taagaaaagt
     6241 attaagtatg ccacaagtaa atatagtaaa aaaaacagaa gtacaaactg gaggatttag
     6301 taaagaaagt atacttccaa aaagaaattc agataaactt attgcaagaa aaaaagattg
     6361 ggatccaaaa aaatatggtg gatttgatag tccaacagta gcatatagtg tattagtagt
     6421 agcaaaagta gaaaaaggta aatctaaaaa acttaaaagt gtaaaagaat tacttggaat
     6481 aacaataatg gaaagaagta gttttgaaaa aaatcctatt gattttttag aagcaaaagg
     6541 atataaagaa gttaaaaaag atttaataat aaaacttcca aaatattctt tatttgaatt
     6601 agaaaatgga agaaaaagaa tgttagcaag tgctggtgaa ttacaaaaag gtaatgaatt
     6661 agcattacca tctaaatatg taaattttct ttatttagca agtcattatg aaaaacttaa
     6721 aggatctcct gaagataatg aacaaaaaca attatttgta gaacaacata aacattattt
     6781 agatgaaata attgaacaaa taagtgaatt ttctaaaaga gtaatattag cagacgcaaa
     6841 tttagataaa gtattatctg cttataataa acatagagat aaaccaataa gagaacaagc
     6901 agaaaatata atacatcttt ttacattaac aaatttaggt gcaccagcag catttaaata
     6961 ttttgataca acaatagata gaaaaagata tacaagtaca aaagaagtat tagacgcaac
     7021 attaatacat caaagtataa caggattata tgaaacaaga atagatttat ctcaattagg
     7081 tggtgattaa aagcttggca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     7141 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     7201 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgct
     7261 agcataaaaa taagaagcct gcatttgcag gcttcttatt tttatggcgc gcccgccctt
     7321 aagtctaaaa attaggggag atgtaaggat ttgggaaaaa tagaagatgt tataatcata
     7381 aatatggtat tcgtaggctt aaagtcaaaa aggaggtgaa atataaatag atttttagct
     7441 aaattaagta agaaatagga ggagatttat tgaacaaaaa attagaaaaa ccatttgtat
     7501 ataagagaga gtacgatttg actggatatg atgttgaaat tttacaaaaa tatgagttag
     7561 aacaagcaat atatgtttat gttgggagta gttgtgcata taacatgaga gctagaagta
     7621 gtaaatggag ataccatata agaacaaata ataagtctat atgttgtaac attaaaaatt
     7681 ttatacataa cttggaattg ttttataaaa tggaattaaa gttgtcagat aatattatta
     7741 atgataagct atactatagc aatatagcag agtttgaaga atttgaaaca ctagaaaaag
     7801 ctagagaggt agaaagtact attataagtc aatatcaatt tttagattct ataaatcaca
     7861 tgttaaaaca aaaaataatt ttattgagta ataaggatag tgtgttaaac ataactaaaa
     7921 atggaaatac aaattatttg aaagtaaaaa ataaatacat agaaaaacat aagaacaagc
     7981 caataatgag ataccatatc aactgtcaat tcaatacaga tggaagtgtc aaaagtatta
     8041 cacaggagtt tgaaccaata ttggaattaa acaaaaaaaa taccctaagc cgaccaagca
     8101 gagtattttt aaaataatat tttaagataa caacaaaatg agataatact actagacaat
     8161 gacaactcaa ctaccaattg agtttatgga gctaccaact ccaatatcgg tctaactgat
     8221 taagtatctg tagttatata ataatattgc tatcaatttt agcatcttaa caatattatt
     8281 atacatacta agctaaaatt attcaatagt tgtaaaagtt gattagtcaa taagtatata
     8341 tttaatgtag tgttatctct taaaaaaact agataaggag ataataaata tatggaacaa
     8401 ttagattcaa aatataagtt gaaaaaattt ctaatggcag tatttagaga tggtatagga
     8461 caaggaaata atcttattga taatgaatat gttagagtat ttcaaaataa taaaagtaat
     8521 agtaaacaat tagaactcgg agaagaattt aaagaatata gtaaaacaac tttttttaaa
     8581 aatatagatg atatagtaga atttaccttc gcaaaaaata tttattatga aaatacattt
     8641 tttaacctat gtactactga tggaaaagca ggaaccaatg aaaacttaat aaatagatat
     8701 gcattaggat ttgattttga caaaaaagaa ttaggacaag gttttaatta taaagatata
     8761 attaatttat ttactaagat aggattacat tatcatatcc tagttgatag tggaaatgga
     8821 ttccatgttt atgtgctaat taataaaact aataacatta agttagtatc agaagttaca
     8881 aatacattaa taaataaatt gggtgcagat aaacaagcaa atttatctac tcaagtatta
     8941 agagtacctt atacatataa tattaaaaat actactaaac aagtaaaaat aatacaccaa
     9001 gacaaaaata tatatagata tgacatagaa aagttagcta aaaaatattg caaagatgta
     9061 aaaacagtag gtaatactaa tacaaaatat atattagata gtaagctacc aaattgtata
     9121 gtagatattt taaaaaatgg tagtaaagat ggacataaaa acctagattt gcaaaaaata
     9181 gttgtgactt taagattgag gaataaaagt ttaagtcaag taatatccgt tgctagagaa
     9241 tggaactata tatcacaaaa tagtctttca aatagtgagc tagaatatca agtcaagtat
     9301 atgtatgaga aacttaaaac ggttaatttt ggttgtactg gttgtgagtt taatagtgat
     9361 tgttggaata aaatagaatc agattttata tatagtgatg aagatacttt gttcaatatg
     9421 ccacataagc actcaaagga tttgaaatat aagaatagga aaggggttaa aataatgact
     9481 ggtaatcaat tgtttatcta taatgtgtta cttaacaata aagatagaga attaaacata
     9541 gacgatataa tggagctgat aacctataaa cgtaagaaga aagttaaaaa cattgttatg
     9601 agtgaaaaga cattaagaga aacattaaaa gaacttcaac ataatgatta tattacaaaa
     9661 acaaaaggtg ttacaaagct aggaataaaa gatacataca atgtaaaaga agttagatgt
     9721 aatatagata aacaatatac tattagttac tttgttacca tggcagtaat ttggggaata
     9781 atttcaactg aagaattaag attatatact cacatgagat ataagcaaga tttattggtc
     9841 aaagatgata aaataaaagg aaatatatta agaattaatc aagaggaatt agcaaaagat
     9901 ttaggagtaa cacagcaaag aatttcaaat atgatagaat ctttattaga tactaaaatt
     9961 ttagatgtat gggaaactaa aataaatgat agaggattta tgtactatac atatagatta
    10021 aacaagtaga tttttgatag gattagaatt gattttctag tcctattttt atgcaaaaaa
    10081 actaattaat aaaaatttct tttggtaaaa taattgtacg agaattgcaa aaaaaaaatg
    10141 gcatcaaagt attgaaatta agccgtttta aaaatttctt ttggtaaaat aattctacat
    10201 atatatgtag tatatatata tatgtttttt agagaatgta taactagaat atagagctag
    10261 aatatagaga atgtataact agaatataga gctagaatat agagaatgta taactagaat
    10321 atagagctag aatatagaga atgtataact agaatataga gctagaatat agagaatgta
    10381 taactagaat atagagctag aatatagaga atgtataact agaatataga gctagaatat
    10441 agagaatgta taactagaat atagagctag aatcctaatt agtaggtgct tttttaaaac
    10501 aagttaaaaa tcaaaaatag tattagtaag cattggaaat gctagattct aaaatagaaa
    10561 agtaaaaaat tggtgcacta tctaaactta tctatatcgc tttttccgtc gtttggttct
    10621 ctagttacga tacaggggat atgcttatat tgagttatag tactaatcag tgcttaatat
    10681 agttaataaa attatagtta ccatagttta gtaactatga tgtatgttag ttagaaactt
    10741 gcatttcggc cggccagtgg gcaagttgaa aaattcacaa aaatgtggta taatatcttt
    10801 gttcattaga gcgataaact tgaatttgag agggaactta gatggtattt gaaaaaattg
    10861 ataaaaatag ttggaacaga aaagagtatt ttgaccacta ctttgcaagt gtaccttgta
    10921 cctacagcat gaccgttaaa gtggatatca cacaaataaa ggaaaaggga atgaaactat
    10981 atcctgcaat gctttattat attgcaatga ttgtaaaccg ccattcagag tttaggacgg
    11041 caatcaatca agatggtgaa ttggggatat atgatgagat gataccaagc tatacaatat
    11101 ttcacaatga tactgaaaca ttttccagcc tttggactga gtgtaagtct gactttaaat
    11161 catttttagc agattatgaa agtgatacgc aacggtatgg aaacaatcat agaatggaag
    11221 gaaagccaaa tgctccggaa aacattttta atgtatctat gataccgtgg tcaaccttcg
    11281 atggctttaa tctgaatttg cagaaaggat atgattattt gattcctatt tttactatgg
    11341 ggaaatatta taaagaagat aacaaaatta tacttccttt ggcaattcaa gttcatcacg
    11401 cagtatgtga cggatttcac atttgccgtt ttgtaaacga attgcaggaa ttgataaata
    11461 gttaacttca ggtttgtctg taactaaaaa caagtattta agcaaaaaca tcgtagaaat
    11521 acggtgtttt ttgttaccct aagtttcata aaaataagaa gcctgcaaat gcaggcttct
    11581 tatttttatg gtttaaacgg ttttagctgg gatatcggca aaacaagcaa atactactat
    11641 agaagcagtt aaaagggcta ttttttataa tctggagcat agacaagaag ttagagaagc
    11701 aattaaaaaa gaaaatataa gaaaaattct tattattggg acatcaaaaa aaatggttaa
    11761 tcaaattgta gagagattgt caattggaag ggtctgtaag tttataaata tacaagatat
    11821 aagtactaaa tatgaaatag aaatagcaaa acaagctaga aaagaaggaa atcatgtaat
    11881 tcctgttcca gccgttgaga taaaaagtat ggcaagtggt ctcagtataa attctttgaa
    11941 gagacttttt agaaaaggaa ataataggaa tatggtagtt ttagaaaaaa caattataag
    12001 accaactttt agttatatag gaaagtttta tattagtgct gatgtaatta aacagattat
    12061 tgaatatgaa gtatacaact ttggaagcat tgatagaata aataaaatca atatagagaa
    12121 ttgcaataat tttatgaata tttttgtaag tataaatatc aatgatttag aaactataaa
    12181 aaaaattgaa gacatacaac agacaataaa aaagtctatt gagaaaatga ctttagttaa
    12241 tgtacaaaaa atagatatta acattcataa aattaaaaaa ataaacttcc tccaatgtta
    12301 aaaaataaac tgagaaaatg atatactaat ttgcaataaa gtgtgctata attaaactgt
    12361 aaaggtaccg aaaagtgatg cattgttggg ttttagagct agaaatagca agttaaaata
    12421 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt ttctatggag
    12481 aaatctagat cagcatgatg tctgactaga cgcgtaagct ctgcaactat ttttagatgg
    12541 ttgcagagct tatttttatt ttttattaaa ctcctttttg ataatctcat gaccaaaatc
    12601 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
    12661 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
    12721 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
    12781 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
    12841 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
    12901 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
    12961 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
    13021 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
    13081 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
    13141 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
    13201 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
    13261 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
    13321 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
    13381 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
    13441 atacgcaggg ccctaacatc ttctattttt cccaaatcct tacatctccc ctaattttta
    13501 gacttaaggg cgaattc
//
