LOCUS       sequence-390787-        9116 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid for dCas9 in E. coli used in Chip-seq 
            experiments. dCas9 is tagged with a C-terminal 3x FLAG tag. A guide 
            RNA can be cloned using BsaI restriction sites..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9116)
  AUTHORS   Rostain W, Grebert T, Vyhovskyi D, Pizarro PT, Tshinsele-Van 
            Bellingen G, Cui L, Bikard D
  TITLE     Cas9 off-target binding to the promoter of bacterial genes leads to 
            silencing and toxicity.
  JOURNAL   Nucleic Acids Res. 2023 Mar 17:gkad170. doi: 10.1093/nar/gkad170.
  PUBMED    36929199
REFERENCE   2  (bases 1 to 9116)
  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: "Nucleic 
            Acids Res. 2023 Mar 17:gkad170. doi: 10.1093/nar/gkad170."
COMMENT     Sequence Label: pLC133
FEATURES             Location/Qualifiers
     source          1..9116
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(69..1652,1653..1655,1656..4142,4143..4145,
                     4146..4172))
                     /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"
     CDS             4374..5000
                     /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     complement(4450..4469)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      5037..5123
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        5760..5862
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             5863..6522
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(5929..5948)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      7226..7448
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             7496..8446
                     /codon_start=1
                     /gene="rep101 encoding an A56V mutation"
                     /product="temperature-sensitive version of the RepA protein
                     needed for replication with the pSC101 origin (Armstrong et
                     al., 1984)"
                     /label=Rep101(Ts)
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     protein_bind    8589..8607
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8614..8632
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_RNA        8663..8738
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      complement(8782..9026)
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     CDS             complement(join(9107..9116,1..56))
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
ORIGIN
        1 tcatccttgt aatcgatgtc atgatcttta taatcaccgt catggtcttt gtagtcaccg
       61 ctaccgccgt cacctcctag ctgactcaaa tcaatgcgtg tttcataaag accagtgatg
      121 gattgatgga taagagtggc atctaaaact tcttttgtag acgtatatcg tttacgatca
      181 attgttgtat caaaatattt aaaagcagcg ggagctccaa gattcgtcaa cgtaaataaa
      241 tgaataatat tttctgcttg ttcacgtatt ggtttgtctc tatgtttgtt atatgcacta
      301 agaactttat ctaaattggc atctgctaaa ataacacgct tagaaaattc actgatttgc
      361 tcaataatct catctaaata atgcttatgc tgctccacaa acaattgttt ttgttcgtta
      421 tcttctggac tacccttcaa cttttcataa tgactagcta aatataaaaa attcacatat
      481 ttgcttggca gagccagctc atttcctttt tgtaattctc cggcactagc cagcatccgt
      541 ttacgaccgt tttctaactc aaaaagacta tatttaggta gtttaatgat taagtctttt
      601 ttaacttcct tatatccttt agcttctaaa aagtcaatcg gatttttttc aaaggaactt
      661 ctttccataa ttgtgatccc tagtaactct ttaacggatt ttaacttctt cgatttccct
      721 ttttccacct tagcaaccac taggactgaa taagctaccg ttggactatc aaaaccacca
      781 tatttttttg gatcccagtc ttttttacga gcaataagct tgtccgaatt tctttttggt
      841 aaaattgact ccttggagaa tccgcctgtc tgtacttctg ttttcttgac aatattgact
      901 tggggcatgg acaatacttt gcgcactgtg gcaaaatctc gccctttatc ccagacaatt
      961 tctccagttt ccccattagt ttcgattaga gggcgtttgc gaatctctcc atttgcaagt
     1021 gtaatttctg ttttgaagaa gttcatgata ttagagtaaa agaaatattt tgcggttgct
     1081 ttgcctattt cttgctcaga cttagcaatc attttacgaa catcataaac tttataatca
     1141 ccatagacaa actccgattc aagttttgga tatttcttaa tcaaagcagt tccaacgacg
     1201 gcatttagat acgcatcatg ggcatgatgg taattgttaa tctcacgtac tttatagaat
     1261 tggaaatctt ttcggaagtc agaaactaat ttagatttta aggtaatcac tttaacctct
     1321 cgaataagtt tatcattttc atcgtattta gtattcatgc gactatccaa aatttgtgcc
     1381 acatgcttag tgatttggcg agtttcaacc aattggcgtt tgataaaacc agctttatca
     1441 agttcactca aacctccacg ttcagctttc gttaaattat caaacttacg ttgagtgatt
     1501 aacttggcgt ttagaagttg tctccaatag tttttcatct ttttgactac ttcttcactt
     1561 ggaacgttat ccgatttacc acgattttta tcagaacgcg ttaagacctt attgtctatt
     1621 gaatcgtctt taaggaaact ttgtggaaca atggcatcga catcataatc acttaaacga
     1681 ttaatatcta attcttggtc cacatacatg tctcttccat tttggagata atagagatag
     1741 agcttttcat tttgcaattg agtattttca acaggatgct ctttaagaat ctgacttcct
     1801 aattctttga taccttcttc gattcgtttc atacgctctc gcgaattttt ctggcccttt
     1861 tgagttgtct gattttcacg tgccatttca ataacgatat tttctggctt atgccgcccc
     1921 attactttga ccaattcatc aacaactttt acagtctgta aaataccttt tttaatagca
     1981 gggctaccag ctaaatttgc aatatgttca tgtaaactat cgccttgtcc agacacttgt
     2041 gctttttgaa tgtcttcttt aaatgtcaaa ctatcatcat ggatcagctg cataaaattg
     2101 cgattggcaa aaccatctga tttcaaaaaa tctaatattg ttttgccaga ttgcttatcc
     2161 ctaataccat taatcaattt tcgagacaaa cgtccccaac cagtataacg gcgacgttta
     2221 agctgtttca tcaccttatc atcaaagagg tgagcatatg ttttaagtct ttcctcaatc
     2281 atctccctat cttcaaataa ggtcaatgtt aaaacaatat cctctaagat atcttcattt
     2341 tcttcattat ccaaaaaatc tttatcttta ataattttta gcaaatcatg gtaggtacct
     2401 aatgaagcat taaatctatc ttcaactcct gaaatttcaa cactatcaaa acattctatt
     2461 tttttgaaat aatcttcttt taattgctta acggttactt ttcgatttgt tttgaagagt
     2521 aaatcaacaa tggctttctt ctgttcacct gaaagaaatg ctggttttcg cattccttca
     2581 gtaacatatt tgacctttgt caattcgtta taaaccgtaa aatactcata aagcaaacta
     2641 tgttttggta gtactttttc atttggaaga tttttatcaa agtttgtcat gcgttcaata
     2701 aatgattgag ctgaagcacc tttatcgaca acttcttcaa aattccatgg ggtaattgtt
     2761 tcttcagact tccgagtcat ccatgcaaaa cgactattgc cacgcgccaa tggaccaaca
     2821 taataaggaa ttcgaaaagt caagattttt tcaatcttct cacgattgtc ttttaaaaat
     2881 ggataaaagt cttcttgtct tctcaaaata gcatgcagct cacccaagtg aatttgatgg
     2941 ggaatagagc cgttgtcaaa ggtccgttgc ttgcgcagca aatcttcacg atttagtttc
     3001 accaataatt cctcagtacc atccattttt tctaaaattg gtttgataaa tttataaaat
     3061 tcttcttggc tagctccccc atcaatataa cctgcatatc cgttttttga ttgatcaaaa
     3121 aagatttctt tatacttttc tggaagttgt tgtcgaacta aagcttttaa aagagtcaag
     3181 tcttgatgat gttcatcgta gcgtttaatc attgaagctg ataggggagc cttagttatt
     3241 tcagtattta ctcttaggat atctgaaagt aaaatagcat ctgataaatt cttagctgcc
     3301 aaaaacaaat cagcatattg atctccaatt tgcgccaata aattatctaa atcatcatcg
     3361 taagtatctt ttgaaagctg taatttagca tcttctgcca aatcaaaatt tgatttaaaa
     3421 ttaggggtca aacccaatga caaagcaatg agattcccaa ataagccatt tttcttctca
     3481 ccggggagct gagcaatgag attttctaat cgtcttgatt tactcaatcg tgcagaaaga
     3541 atcgctttag catctactcc acttgcgtta atagggtttt cttcaaataa ttgattgtag
     3601 gtttgtacca actggataaa tagtttgtcc acatcactat tatcaggatt taaatctccc
     3661 tcaatcaaaa aatgaccacg aaacttaatc atatgcgcta aggccaaata gattaagcgc
     3721 aaatccgctt tatcagtaga atctaccaat ttttttcgca gatgatagat agttggatat
     3781 ttctcatgat aagcaacttc atctactata tttccaaaaa taggatgacg ttcatgcttc
     3841 ttgtcttctt ccaccaaaaa agactcttca agtcgatgaa agaaactatc atctactttc
     3901 gccatctcat ttgaaaaaat ctcctgtaga taacaaatac gattcttccg acgtgtatac
     3961 cttctacgag ctgtccgttt gagacgagtc gcttccgctg tctctccact gtcaaataaa
     4021 agagccccta taagattttt tttgatactg tggcggtctg tatttcccag aaccttgaac
     4081 tttttagacg gaaccttata ttcatcagtg atcaccgccc atccgacgct atttgtgccg
     4141 atagctaagc ctattgagta tttcttatcc atttttgcct cctaactagg tcatttgata
     4201 tgcctccgga tatcactcta tcaatgatag agagcttatt ttaattatgc tctatcaatg
     4261 atagagtgtc aatatttttt ttagtttttc atgaactcga ggggatccaa ataaaaaact
     4321 agtttgacaa ataactctat caatgatata atgtcaacaa aaaggaggaa ttaatgatgt
     4381 ctagattaga taaaagtaaa gtgattaaca gcgcattaga gctgcttaat gaggtcggaa
     4441 tcgaaggttt aacaacccgt aaactcgccc agaagctagg tgtagagcag cctacattgt
     4501 attggcatgt aaaaaataag cgggctttgc tcgacgcctt agccattgag atgttagata
     4561 ggcaccatac tcacttttgc cctttagaag gggaaagctg gcaagatttt ttacgtaata
     4621 acgctaaaag ttttagatgt gctttactaa gtcatcgcga tggagcaaaa gtacatttag
     4681 gtacacggcc tacagaaaaa cagtatgaaa ctctcgaaaa tcaattagcc tttttatgcc
     4741 aacaaggttt ttcactagag aatgcattat atgcactcag cgctgtgggg cattttactt
     4801 taggttgcgt attggaagat caagagcatc aagtcgctaa agaagaaagg gaaacaccta
     4861 ctactgatag tatgccgcca ttattacgac aagctatcga attatttgat caccaaggtg
     4921 cagagccagc cttcttattc ggccttgaat tgatcatatg cggattagaa aaacaactta
     4981 aatgtgaaag tgggtcttaa ccgatcccca attcctggca ggcgtgctag aggcatcaaa
     5041 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
     5101 acgctctcct gagtaggaca aatccgccgc cctagaccct ccacgcacgt tgtgatatgt
     5161 agatgataat cattatcact ttacgggtcc tttccggtga tccgacaggt tacggggcgg
     5221 cgacctcgcg ggttttcgct atttatgaaa attttccggt ttaaggcgtt tccgttcttc
     5281 ttcgtcataa cttaatgttt ttatttaaaa taccctctga aaagaaagga aacgacaggt
     5341 gctgaaagcg aggctttttg gggcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5401 gcggtaatac ggttatccac agaatcaggg gataacgcag gcctccgaaa ttagtttaat
     5461 atgctgccaa catagtaagc cagtatacac tccgctagcg ctgatgtccg gcggtgcttt
     5521 tgccgttacg caccaccccg tcagtagctg aacaggaggg acagctgata gaaacagaag
     5581 ccactggagc acctcaaaaa caccatcata cactaaatca gtaagttggc agcatcaccc
     5641 gacgcacttt gcgccgaata aatacctgtg acggaagatc acttcgcaga ataaataaat
     5701 cctggtgtcc ctgttgatac cgggaagccc cgggccaact tttggcgaaa atgagacgtt
     5761 gatcggcacg taagaggttc caactttcac cataatgaaa taagatcact accgggcgta
     5821 ttttttgagt tatcgagatt ttcaggagct aaggaagcta aaatggagaa aaaaatcact
     5881 ggatatacca ccgttgatat atcccaatgg catcgtaaag aacattttga ggcatttcag
     5941 tcagttgctc aatgtaccta taaccagacc gttcagctgg atattacggc ctttttaaag
     6001 accgtaaaga aaaataagca caagttttat ccggccttta ttcacattct tgcccgcctg
     6061 atgaatgctc atccggaatt ccgtatggca atgaaagacg gtgagctggt gatatgggat
     6121 agtgttcacc cttgttacac cgttttccat gagcaaactg aaacgttttc atcgctctgg
     6181 agtgaatacc acgacgattt ccggcagttt ctacacatat attcgcaaga tgtggcgtgt
     6241 tacggtgaaa acctggccta tttccctaaa gggtttattg agaatatgtt tttcgtctca
     6301 gccaatccct gggtgagttt caccagtttt gatttaaacg tggccaatat ggacaacttc
     6361 ttcgcccccg ttttcaccat gggcaaatat tatacgcaag gcgacaaggt gctgatgccg
     6421 ctggcgattc aggttcatca tgccgtttgt gatggcttcc atgtcggcag aatgcttaat
     6481 gaattacaac agtactgcga tgagtggcag ggcggggcgt aattttttta aggcagttat
     6541 tggtgccctt aaacgcctgg ttgctacgcc tgaataagtg ataataagcg gatgaatggc
     6601 ataactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     6661 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     6721 tgagttttcg ttccactgag cgtcagaccc cgttgatgat accgctgcct tactgggtgc
     6781 attagccagt ctgaatgacc tgtcacggga taatccgaag tggtcagact ggaaaatcag
     6841 agggcaggaa ctgctgaaca gcaaaaagtc agatagcacc acatagcaga cccgccataa
     6901 aacgccctga gaagcccgtg acgggctttt cttgtattat gggtagtttc cttgcatgaa
     6961 tccataaaag gcgcctgtag tgccatttac ccccattcac tgccagagcc gtgagcgcag
     7021 cgaactgaat gtcacgaaaa agacagcgac tcaggtgcct gatggtcgga gacaaaagga
     7081 atattcagcg atttgcccga ttgcggccgc aaccgagctt gcgagggtgc tacttaagcc
     7141 tttagggttt taaggtctgt tttgtagagg agcaaacagc gtttgcgaca tccttttgta
     7201 atactgcgga actgactaaa gtagtgagtt atacacaggg ctgggatcta ttctttttat
     7261 ctttttttat tctttcttta ttctataaat tataaccact tgaatataaa caaaaaaaac
     7321 acacaaaggt ctagcggaat ttacagaggg tctagcagaa tttacaagtt ttccagcaaa
     7381 ggtctagcag aatttacaga tacccacaac tcaaaggaaa aggactagta attatcattg
     7441 actagcccat ctcaattggt atagtgatta aaatcaccta gaccaattga gatgtatgtc
     7501 tgaattagtt gttttcaaag caaatgaact agcgattagt cgctatgact taacggagca
     7561 tgaaaccaag ctaattttat gctgtgtggc actactcaac cccacgattg aaaaccctac
     7621 aaggaaagaa cggacggtat cgttcactta taaccaatac gttcagatga tgaacatcag
     7681 tagggaaaat gcttatggtg tattagctaa agcaaccaga gagctgatga cgagaactgt
     7741 ggaaatcagg aatcctttgg ttaaaggctt tgagattttc cagtggacaa actatgccaa
     7801 gttctcaagc gaaaaattag aattagtttt tagtgaagag atattgcctt atcttttcca
     7861 gttaaaaaaa ttcataaaat ataatctgga acatgttaag tcttttgaaa acaaatactc
     7921 tatgaggatt tatgagtggt tattaaaaga actaacacaa aagaaaactc acaaggcaaa
     7981 tatagagatt agccttgatg aatttaagtt catgttaatg cttgaaaata actaccatga
     8041 gtttaaaagg cttaaccaat gggttttgaa accaataagt aaagatttaa acacttacag
     8101 caatatgaaa ttggtggttg ataagcgagg ccgcccgact gatacgttga ttttccaagt
     8161 tgaactagat agacaaatgg atctcgtaac cgaacttgag aacaaccaga taaaaatgaa
     8221 tggtgacaaa ataccaacaa ccattacatc agattcctac ctacataacg gactaagaaa
     8281 aacactacac gatgctttaa ctgcaaaaat tcagctcacc agttttgagg caaaattttt
     8341 gagtgacatg caaagtaagt atgatctcaa tggttcgttc tcatggctca cgcaaaaaca
     8401 acgaaccaca ctagagaaca tactggctaa atacggaagg atctgaggtt cttatggctc
     8461 ttgtatctat cagtgaagca tcaagactaa caaacaaaag tagaacaact gttcaccgtt
     8521 acatatcaaa gggaaaactg tccataccca tggaaggcga gcggcatgca tttacgttga
     8581 caccatcgtc cctatcagtg atagagattg acatccctat cagtgataga gatactgagc
     8641 actgagacca gtctaggtct cggttttaga gctagaaata gcaagttaaa ataaggctag
     8701 tccgttatca acttgaaaaa gtggcaccga gtcggtgctt tttttgagat ctgtccatac
     8761 ccatggattc ttcgtctgtt tctactggta ttggcacaaa cctgattcca atttgagcaa
     8821 ggctatgtgc catctcgata ctcgttctta actcaacaga agatgctttg tgcatacagc
     8881 ccctcgttta ttatttatct cctcagccag ccgctgtgct ttcagtggat ttcggataac
     8941 agaaaggccg ggaaataccc agcctcgctt tgtaacggag tagacgaaag tgattgcgcc
     9001 tacccggata ttatcgtgag gatgcgtcat cagtttaggt taggcgccat gcatctcgag
     9061 gcatgcctgc aatgcctgga gatccttact cgagtcatcc tgcagccttg tcatcg
//
