LOCUS       sequence-461406-       13309 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  optimized ahTet inducible dCas9 for insertion into C. difficile 
            genome.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13309)
  AUTHORS   Chua M, Erickson D, Collins J
  TITLE     Plasmid sequences and availability of a two-plasmid system for 
            CRISPRi knockdown of Clostridioides difficile genes without 
            antibiotic selection.
  JOURNAL   Microbiol Resour Announc. 2025 Apr 28:e0011625. doi: 
            10.1128/mra.00116-25.
  PUBMED    40293262
REFERENCE   2  (bases 1 to 13309)
  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: "Microbiol 
            Resour Announc. 2025 Apr 28:e0011625. doi: 10.1128/mra.00116-25."
COMMENT     Sequence Label: pJAK184.tetR.PT5-3.dCas9
FEATURES             Location/Qualifiers
     source          1..13309
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      91..679
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     580..599
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     833..850
                     /label=L4440
                     /note="L4440 vector, forward primer"
     oriT            1000..1109
                     /note="incP origin of transfer"
     CDS             1142..1513
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     primer_bind     1710..1726
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1710..1726
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3561..3578)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3561..3577)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3570..3592)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(7643..8269)
                     /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     8174..8193
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     protein_bind    8389..8407
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    8414..8432
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     CDS             join(8461..8487,8488..8490,8491..10977,10978..10980,
                     10981..12567)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
ORIGIN
        1 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
       61 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
      121 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
      181 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
      241 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
      301 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
      361 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
      421 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
      481 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
      541 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
      601 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
      661 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
      721 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
      781 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
      841 agtgagcgag gaagcggaag agcgcccaat acgcagggcc ccctgcttcg gggtcattat
      901 agcgattttt tcggtatatc catccttttt cgcacgatat acaggatttt gccaaagggt
      961 tcgtgtagac tttccttggt gtatccaacg gcgtcagccg ggcaggatag gtgaagtagg
     1021 cccacccgcg agcgggtgtt ccttcttcac tgtcccttat tcgcacctgg cggtgctcaa
     1081 cgggaatcct gctctgcgag gctggccggc taccgccggc gtaacagatg agggcaagcg
     1141 gatggctgat gaaaccaagc caaccaggaa gggcagccca cctatcaagg tgtactgcct
     1201 tccagacgaa cgaagagcga ttgaggaaaa ggcggcggcg gccggcatga gcctgtcggc
     1261 ctacctgctg gccgtcggcc agggctacaa aatcacgggc gtcgtggact atgagcacgt
     1321 ccgcgagctg gcccgcatca atggcgacct gggccgcctg ggcggcctgc tgaaactctg
     1381 gctcaccgac gacccgcgca cggcgcggtt cggtgatgcc acgatcctcg ccctgctggc
     1441 gaagatcgaa gagaagcagg acgagcttgg caaggtcatg atgggcgtgg tccgcccgag
     1501 ggcagagcca tgactttttt agccgctaaa acggccgggg ggtgcgcgtg attgccaagc
     1561 acgtccccat gcgctccatc aagaagagcg acttcgcgga gctggtgaag tacatcaccg
     1621 acgagcaagg caagaccgat cgggccccct gcaggataaa aaaattgtag ataaatttta
     1681 taaaatagtt ttatctacaa tttttttatc aggaaacagc tatgaccgcg gccgcgctgt
     1741 atatagtcat aacccttata tttcattaat taaagttaaa ttttgtatat tcaaaaagct
     1801 cttgatattt acggcagagc caccatatag agatgcctca gaccctaatg atgaaatttc
     1861 tatatttatt cctttgttaa aagagctaac taacatatct tttgtaatct gtaatatctc
     1921 agggatatca ctaattattt ggctatttaa atatattatt tcaggagcat aagttgtaat
     1981 agcattattt atagctattg ttaagtaact acaaaactca tgaataactt ttttggcatt
     2041 ctggttatct tcataatata gttgctttac tatatcagaa tctattttag gaatattttc
     2101 taaagaagaa agctgttcaa ataccttttt ttctgaacaa tactgttcta aacagccacg
     2161 atttccacaa ggacatagtt taccattagg catgatgata gtgtgaccaa tttctccact
     2221 cataccattt ctgccgctat ataatttatt atttattata atgcctgaac caaatccact
     2281 gtgaatactg agactaagta gtgagttatg tacagttgaa aaagtattct cagctaaagc
     2341 tgttaagttt gcttcatttt ctatatgaat aggaaagtca tactttttgc taaggatgct
     2401 atataagtca atctcattaa gattataata tggagtaaac aatactttat tttcacaagt
     2461 aatcccatgt atagccaaag taagaccaat taccttataa ggagtgtcta ttttggatat
     2521 gttataacta tttataattt catctatcaa ttgtataaca ttgtctttac ttacttgtat
     2581 atctgttaat ttcttagaat ttataatagt tccatctaag taagatagag aagaaaatat
     2641 atagtcgtat ccaatgtcca tacttaaaga aatccctgca catttattaa atactaataa
     2701 tatgggtttt cttccgccac tatgagtact atttccaatt cctatttcat gaactagaga
     2761 ttcatcaata agttttttgg taatagcgga tatagttgct ttattcaatc ctatagtaga
     2821 agctatactt gccctagaaa taggaccatt ttttataatt tgttcaagta ccaatctttc
     2881 attcatttct cgaatagtgt atttatcagt aaccaatttg atattcctcc ttaaaataat
     2941 attgtaatac tttttacaca aaaataaaag gttattttgc attgacaaag ataattaaat
     3001 attttattat tagttcataa gttagtttaa tatactaaca aaaataaagc aagtaaaata
     3061 tacctaaaat ataaaaaaat taggatagga aaacgatagt tatgaagtgg cgcagtaaag
     3121 gagaaaattt tatggtaagg aaaacgatag ttatgaagtg gcgcagtaaa ggagaaaatt
     3181 ttatggtaag tagatatgta ccagatatgg gagatttaat atgggttgat tttgatccaa
     3241 caaaaggtag tgaacaagca ggacatagac cagctgttgt attaagtcca tttatgtata
     3301 ataataaaac aggtatgtgt ttatgtgttc cttgtacaac tcaatcaaaa ggatatccat
     3361 ttgaagttgt tttatcaggt caagaaagag atggagtagc attagctgat caagtaaaaa
     3421 gtatagcatg gagagcaaga ggagcaacta aaaaaggaac agttgcacca gaagaattac
     3481 aattaattaa agcaaaaatt aatgtattaa ttggataaat caacaactct cctcgaggcc
     3541 tgcagacatg caagcttggc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     3601 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     3661 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc
     3721 tagcataaaa ataagaagcc tgcatttgca ggcttcttat ttttatggcg cgccgttctg
     3781 aatccttagc taatggttca acaggtaact atgacgaaga tagcaccctg gataagtctg
     3841 taatggattc taaggcattt aatgaagacg tgtatataaa atgtgctaat gaaaaagaaa
     3901 atgcgttaaa agagcctaaa atgagttcaa atggttttga aattgattgg tagtttaatt
     3961 taatatattt tttctattgg ctatctcgat acctatagaa tcttctgttc acttttgttt
     4021 ttgaaatata aaaaggggct ttttagcccc ttttttttaa aactccggag gagtttcttc
     4081 attcttgata ctatacgtaa ctatttcgat ttgacttcat tgtcaattaa gctagtaaaa
     4141 tcaatggtta aaaaacaaaa aacttgcatt tttctaccta gtaatttata attttaagtg
     4201 tcgagtttaa aagtataatt taccaggaaa ggagcaagtt ttttaataag gaaaaatttt
     4261 tccttttaaa attctatttc gttatatgac taattataat caaaaaaatg aaaataaaca
     4321 agaggtaaaa actgctttag agaaatgtac tgataaaaaa agaaaaaatc ctagatttac
     4381 gtcatacata gcacctttaa ctactaagaa aaatattgaa aggacttcca cttgtggaga
     4441 ttatttgttt atgttgagtg atgcagactt agaacatttt aaattacata aaggtaattt
     4501 ttgcggtaat agattttgtc caatgtgtag ttggcgactt gcttgtaagg atagtttaga
     4561 aatatctatt cttatggagc atttaagaaa agaagaaaat aaagagttta tatttttaac
     4621 tcttacaact ccaaatgtaa aaagttatga tcttaattat tctattaaac aatataataa
     4681 atcttttaaa aaattaatgg agcgtaagga agttaaggat ataactaaag gttatataag
     4741 aaaattagaa gtaacttacc aaaaggaaaa atacataaca aaggatttat ggaaaataaa
     4801 aaaagattat tatcaaaaaa aaggacttga aattggtgat ttagaaccta attttgatac
     4861 ttataatcct cattttcatg tagttattgc agttaataaa agttatttta cagataaaaa
     4921 ttattatata aatcgagaaa gatggttgga attatggaag tttgctacta aggatgattc
     4981 tataactcaa gttgatgtta gaaaagcaaa aattaatgat tataaagagg tttacgaact
     5041 tgcgaaatat tcagctaaag acactgatta tttaatatcg aggccagtat ttgaaatttt
     5101 ttataaagca ttaaaaggca agcaggtatt agtttttagt ggatttttta aagatgcaca
     5161 caaattgtac aagcaaggaa aacttgatgt ttataaaaag aaagatgaaa ttaaatatgt
     5221 ctatatagtt tattataatt ggtgcaaaaa acaatatgaa aaaactagaa taagggaact
     5281 tacggaagat gaaaaagaag aattaaatca agatttaata gatggaatag aaatagatta
     5341 aagtgtaact atactttata tatatatgat taaaaaaata aaaaacaaca gcctattagg
     5401 ttgttgtttt ttattttctt tattaatttt tttaattttt agtttttagt tcttttttaa
     5461 aataagtttc agcctctttt tcaatatttt ttaaagaagg agtatttgca tgaattgcct
     5521 tttttctaac agacttagga aatattttaa cagtatcttc ttgcgccggt gattttggaa
     5581 cttcataact tactaattta taattattat tttctttttt aattgtaaca gttgcaaaag
     5641 aagctgaacc tgttccttca actagtttat catcttcaat ataatattct tgacctatat
     5701 agtataaata tatttttatt atatttttac ttttttctga atctattatt ttataatcat
     5761 aaaaagtttt accaccaaaa gaaggttgta ctccttctgg tccaacatat ttttttacta
     5821 tattatctaa ataatttttg ggaactggtg ttgtaatttg attaatcgaa caaccagtta
     5881 tacttaaagg aattataact ataaaaatat ataggattat ctttttaaat ttcattattg
     5941 gcctcctttt tattaaattt atgttaccat aaaaaggaca taacgggaat atgtagaata
     6001 tttttaatgt agacaaaatt ttacataaat ataaagaaag gaagtgtttg tttaaatttt
     6061 atagcaaact atcaaaaatt agggggataa aaatttatga aaaaaaggtt ttcgatgtta
     6121 tttttatgtt taactttaat agtttgtggt ttatttacaa attcggccgg ccagtgggca
     6181 agttgaaaaa ttcacaaaaa tgtggtataa tatctttgtt cattagagcg ataaacttga
     6241 atttgagagg gaacttagat ggtatttgaa aaaattgata aaaatagttg gaacagaaaa
     6301 gagtattttg accactactt tgcaagtgta ccttgtacct acagcatgac cgttaaagtg
     6361 gatatcacac aaataaagga aaagggaatg aaactatatc ctgcaatgct ttattatatt
     6421 gcaatgattg taaaccgcca ttcagagttt aggacggcaa tcaatcaaga tggtgaattg
     6481 gggatatatg atgagatgat accaagctat acaatatttc acaatgatac tgaaacattt
     6541 tccagccttt ggactgagtg taagtctgac tttaaatcat ttttagcaga ttatgaaagt
     6601 gatacgcaac ggtatggaaa caatcataga atggaaggaa agccaaatgc tccggaaaac
     6661 atttttaatg tatctatgat accgtggtca accttcgatg gctttaatct gaatttgcag
     6721 aaaggatatg attatttgat tcctattttt actatgggga aatattataa agaagataac
     6781 aaaattatac ttcctttggc aattcaagtt catcacgcag tatgtgacgg atttcacatt
     6841 tgccgttttg taaacgaatt gcaggaattg ataaatagtt aacttcaggt ttgtctgtaa
     6901 ctaaaaacaa gtatttaagc aaaaacatcg tagaaatacg gtgttttttg ttaccctagg
     6961 ggagaagtaa aagttccagt attagggaca ggacttaatc aaagtattat agtggtaaat
     7021 gatgatacat attttaaact aagagataag ggtcaacaag tatattttta tggtgcaaag
     7081 gtatttcaag aagaaaatag ccaagtaatg ttcaacaata ttgaaaagaa tttaagttta
     7141 aaaaatggat atattcagaa tggatataaa gctcaaggtg agagtcaatg gatgaagttt
     7201 gcatatgtag tgcttgtgtt tttattctta gtctttgcaa ttgtagctgg aagtataata
     7261 tatatgaaaa tatatagtga tgcatatgaa gataaagaga aatacaagat tttattaaaa
     7321 ataggtacta cagaaaaaga tataaataga gctatactta aagaagtagc tatattttat
     7381 actttgccaa tgttaagtgc aactatatct agttattttg cattgagatt agcaggagat
     7441 ttattgatga acgatttatc tggtatatat atactaagtt tagtgatatg tttgataata
     7501 ttcgtaatat atggagcagt atctgttaat aagtttaaga gtgtagtgta taaaaattag
     7561 gctaaaacta ttaatcttat catatccgca aaaaaccccg cttcggcggg gttttttcgc
     7621 aattcggtcg acagatcgta ccttaagacc cactttcaca tttaagttgt ttttctaatc
     7681 cgcatatgat caattcaagg ccgaataaga aggctggctc tgcaccttgg tgatcaaata
     7741 attcgatagc ttgtcgtaat aatggcggca tactatcagt agtaggtgtt tccctttctt
     7801 ctttagcgac ttgatgctct tgatcttcca atacgcaacc taaagtaaaa tgccccacag
     7861 cgctgagtgc atataatgca ttctctagtg aaaaaccttg ttggcataaa aaggctaatt
     7921 gattttcgag agtttcatac tgtttttctg taggccgtgt acctaaatgt acttttgctc
     7981 catcgcgatg acttagtaaa gcacatctaa aacttttagc gttattacgt aaaaaatctt
     8041 gccagctttc cccttctaaa gggcaaaagt gagtatggtg cctatctaac atctcaatgg
     8101 ctaaggcgtc gagcaaagcc cgcttatttt ttacatgcca atacaatgta ggctgctcta
     8161 cacctagctt ctgggcgagt ttacgggttg ttaaaccttc gattccgacc tcattaagca
     8221 gctctaatgc gctgttaatc actttacttt tatctaatct agacatcatt aattcctcct
     8281 ttttgttgac attatatcat tgatagagtt atttgtcaaa ctagtttttt atttcgatgc
     8341 cctggacttc atgcttagaa ctcaaccatt ctacagttta ttcttgacac tctatcattg
     8401 atagagtata ataactctat cattgataga gtgagctcct gcagtaaagg agaaaatttt
     8461 atggataaaa aatatagtat aggattagca ataggaacaa atagtgtagg atgggcagta
     8521 ataactgatg aatataaagt accaagtaaa aaatttaaag tattaggaaa tacagataga
     8581 catagtataa aaaaaaattt aataggtgca ttactttttg atagtggtga aacagcagaa
     8641 gcaacaagac ttaaaagaac agcaagaaga agatatacta gaagaaaaaa tagaatatgt
     8701 tatttacaag aaatatttag taatgaaatg gcaaaagtag atgatagttt ttttcataga
     8761 ttagaagaaa gttttttagt agaagaagat aaaaaacacg aaagacatcc aatatttgga
     8821 aatatagtag atgaagtagc atatcatgaa aaatatccaa caatatatca tttaagaaaa
     8881 aaacttgtag atagtacaga taaagcagat ttaagactta tatatcttgc attagcacat
     8941 atgataaaat ttagaggaca ttttcttata gaaggtgatt taaatccaga taatagtgat
     9001 gtagataaac tttttataca attagtacaa acatataatc aattatttga agaaaatcca
     9061 ataaatgcaa gtggtgtaga tgcaaaagca atattaagtg caagattaag taaaagtaga
     9121 agattagaaa atcttatagc acaattacct ggagaaaaga aaaatggatt atttggtaat
     9181 cttatagctt taagtttagg attaacacca aattttaaat caaattttga tcttgcagaa
     9241 gatgcaaaac ttcaattaag taaagataca tatgatgatg atttagataa tttacttgca
     9301 caaataggtg atcaatatgc agatttattt cttgcagcaa aaaatctttc tgatgcaata
     9361 cttttaagtg atatacttag agtaaataca gaaataacaa aagcaccatt atctgcaagt
     9421 atgataaaaa gatatgatga acatcatcaa gatttaacat tacttaaagc attagtaaga
     9481 caacaattac cagaaaaata taaagaaatt ttttttgatc aaagtaaaaa tggatatgca
     9541 ggatatatag atggtggtgc aagtcaagaa gaattttata aatttattaa accaatatta
     9601 gaaaaaatgg atggaacaga agaattatta gtaaaactta atagagaaga tttacttaga
     9661 aaacaaagaa catttgataa tggaagtata ccacatcaaa tacatttagg tgaattacat
     9721 gcaatattaa gaagacaaga agatttttat ccatttctta aagataatag agaaaaaatt
     9781 gaaaaaatac ttacatttag aataccatat tatgtaggac cattagcaag aggaaatagt
     9841 agatttgctt ggatgacaag aaaaagtgaa gaaacaataa caccatggaa ttttgaagaa
     9901 gtagtagata aaggagcatc tgcacaaagt tttatagaaa gaatgacaaa ttttgataaa
     9961 aatcttccaa atgaaaaagt attaccaaaa catagtttat tatatgaata ttttacagta
    10021 tataatgaat taacaaaagt aaaatatgta acagaaggta tgagaaaacc agcattttta
    10081 agtggtgaac aaaaaaaagc tatagtagat ttattattta aaacaaatag aaaagtaaca
    10141 gtaaaacaat taaaagaaga ttattttaaa aaaattgaat gttttgatag tgtagaaata
    10201 tctggtgtag aagatagatt taatgcaagt ttaggaacat atcatgattt acttaaaata
    10261 ataaaggata aagatttttt agataatgaa gaaaatgaag atattttaga agatatagta
    10321 ttaacattaa ctttatttga agatagagaa atgatagaag aaagattaaa aacttatgct
    10381 catttatttg atgataaagt aatgaaacaa cttaaaagaa gaagatatac aggttgggga
    10441 agattaagta gaaaacttat aaatggaata agagataaac aaagtggaaa aacaatatta
    10501 gattttctta aaagtgatgg atttgcaaat agaaatttta tgcaattaat acatgatgat
    10561 agtttaacat ttaaagaaga tattcaaaaa gcacaagtaa gtggacaagg tgatagttta
    10621 catgaacata tagcaaattt agcaggatct ccagcaataa aaaaaggtat attacaaaca
    10681 gtaaaagtag ttgatgaatt agtaaaagta atgggtagac ataaaccaga aaatatagtt
    10741 atagaaatgg caagagaaaa tcaaacaaca caaaaaggac aaaaaaatag tagagaaaga
    10801 atgaaaagaa tagaagaagg tataaaagaa ttaggaagtc aaatacttaa agaacatcca
    10861 gtagaaaata cacaacttca aaatgaaaaa ctttatttat attatttaca aaatggaaga
    10921 gatatgtatg tagatcaaga attagatata aatagattaa gtgattatga tgtagatgca
    10981 atagtaccac aaagttttct taaagatgat tctatagata ataaagtatt aacaagatct
    11041 gataaaaata gaggtaaaag tgataatgta ccaagtgaag aagttgtaaa aaaaatgaaa
    11101 aattattgga gacaattact taatgcaaaa cttataacac aaagaaaatt tgataatctt
    11161 acaaaagcag aaagaggtgg attatctgaa ttagataaag ctggatttat aaaaagacaa
    11221 ttagtagaaa ctagacaaat aacaaaacat gtagcacaaa tattagatag tagaatgaat
    11281 acaaaatatg atgaaaatga taagcttata agagaagtaa aagttataac attaaaaagt
    11341 aaattagtaa gtgattttag aaaagatttt caattttata aagtaagaga aataaataat
    11401 tatcatcatg cacatgatgc atatcttaat gcagtagtag gaacagcatt aataaaaaaa
    11461 tatcctaaat tagaatctga atttgtatat ggagattata aagtatatga tgtaagaaaa
    11521 atgatagcaa aaagtgaaca agaaatagga aaagcaacag caaagtactt tttttattct
    11581 aatataatga atttttttaa gacagaaata actttagcaa atggtgaaat aagaaaaaga
    11641 cctttaatag aaactaatgg tgaaacaggt gaaatagtat gggataaagg tagagatttt
    11701 gcaacagtaa gaaaagtatt aagtatgcca caagtaaata tagtaaaaaa aacagaagta
    11761 caaactggag gatttagtaa agaaagtata cttccaaaaa gaaattcaga taaacttatt
    11821 gcaagaaaaa aagattggga tccaaaaaaa tatggtggat ttgatagtcc aacagtagca
    11881 tatagtgtat tagtagtagc aaaagtagaa aaaggtaaat ctaaaaaact taaaagtgta
    11941 aaagaattac ttggaataac aataatggaa agaagtagtt ttgaaaaaaa tcctattgat
    12001 tttttagaag caaaaggata taaagaagtt aaaaaagatt taataataaa acttccaaaa
    12061 tattctttat ttgaattaga aaatggaaga aaaagaatgt tagcaagtgc tggtgaatta
    12121 caaaaaggta atgaattagc attaccatct aaatatgtaa attttcttta tttagcaagt
    12181 cattatgaaa aacttaaagg atctcctgaa gataatgaac aaaaacaatt atttgtagaa
    12241 caacataaac attatttaga tgaaataatt gaacaaataa gtgaattttc taaaagagta
    12301 atattagcag acgcaaattt agataaagta ttatctgctt ataataaaca tagagataaa
    12361 ccaataagag aacaagcaga aaatataata catcttttta cattaacaaa tttaggtgca
    12421 ccagcagcat ttaaatattt tgatacaaca atagatagaa aaagatatac aagtacaaaa
    12481 gaagtattag acgcaacatt aatacatcaa agtataacag gattatatga aacaagaata
    12541 gatttatctc aattaggtgg tgattaactc gagcaaaaac atcgtagaaa tacgaatctc
    12601 gtatgccgtc ttctgcttgc tcgagaaaaa aaaaccgcgc ccctgacagg gcgcggtttt
    12661 tttttcgtaa tttattactg aacaagaggg caatcgatat attaataaag agaaaggaga
    12721 ttcttgtaat aagagtctcc ttgttgatta actaaataat ttgggtaata aatttatata
    12781 aaatatgtta tgatattaat aattcatggc atagtgaaat attaggataa tggagaatat
    12841 acaaatgtat agaattttat tagtagagga cgatattgat ttatcaaaag agattgcgtt
    12901 agcacttgaa aaatggggat ttaaagtaga attaattgat gattttgagg tagtattaga
    12961 tgaatttatg gataaaaaac cagatgtagt acttttagat gtaaatttac cattatataa
    13021 tggattttac tggtgcgaaa aaataagagc tatttcaaat gtaccattaa tatttttatc
    13081 atcaagagat tctgatatgg atttaataat gggtataaat aatggagcag atgattatat
    13141 taccaaacca ttttctatag acatattggt taccaaaata aatggtataa taagaagagt
    13201 ctataattat agtgattcta atagtatttt gtattgtgaa gatttgatgt ttgatgtggg
    13261 gaaagggatt ataaaacata agtataagga taaaagtata gagttgact
//
