LOCUS Exported 10467 bp ds-DNA circular SYN 10-JUN-2021 DEFINITION To introduce dCas9-fused Tet1-CD and sgRNA2.0 system. ACCESSION . VERSION . KEYWORDS pdCas9-Tet1-CD SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10467) AUTHORS Xu X, Tao Y, Gao X, Zhang L, Li X, Zou W, Ruan K, Wang F, Xu GL, Hu R TITLE A CRISPR-based approach for targeted DNA demethylation. JOURNAL Cell Discov. 2016 May 3;2:16009. doi: 10.1038/celldisc.2016.9. eCollection 2016. PUBMED 27462456 REFERENCE 2 (bases 1 to 10467) AUTHORS . TITLE Direct Submission JOURNAL Exported Jun 10, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10467 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..241 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 1..21 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 172..191 /label=LKO.1 5' /note="Human U6 promoter, forward primer" misc_RNA 285..303 /label=MS2 stem loop /note="stem loop that binds the bacteriophage MS2 coat protein" misc_RNA 355..373 /label=MS2 stem loop /note="stem loop that binds the bacteriophage MS2 coat protein" enhancer 416..701 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer; contains an 18-bp deletion relative to the standard CMV enhancer" promoter 703..980 /label=chicken beta-actin promoter intron 981..1208 /label=hybrid intron /note="hybrid between chicken beta-actin (CBA) and minute virus of mice (MMV) introns (Gray et al., 2011)" CDS 1229..1255 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 1259..1279 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 1289..5389 /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD ATLIHQSITGLYETRIDLSQLGGD" CDS 5393..5413 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" primer_bind complement(7508..7525) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 7514..7721 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" repeat_region 7730..7870 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 7730..7859 /label=AAV2 ITR rep_origin 7945..8400 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(8032..8051) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 8242..8263 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(8417..8436) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 8536..8558 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(8596..8614) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 8682..8786 /gene="bla" /label=AmpR promoter CDS 8787..9647 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(9005..9024) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 9818..10406 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 10307..10326 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag 61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga 121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat 181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga 241 cgaaacaccg ggtcttcgag aagacctgtt ttagagctag gccaacatga ggatcaccca 301 tgtctgcagg gcctagcaag ttaaaataag gctagtccgt tatcaacttg gccaacatga 361 ggatcaccca tgtctgcagg gccaagtggc accgagtcgg tgcttttttg gtacccgtta 421 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt 481 caatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 541 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca 601 atgacggtaa atggcccgcc tggcattgtg cccagtacat gaccttatgg gactttccta 661 cttggcagta catctacgta ttagtcatcg ctattaccat ggtcgaggtg agccccacgt 721 tctgcttcac tctccccatc tcccccccct ccccaccccc aattttgtat ttatttattt 781 tttaattatt ttgtgcagcg atgggggcgg gggggggggg ggggcgcgcg ccrggsgggg 841 sggggsgggg sgrggggsgg ggsggggsgr ggcggagagg tgcggcggca gccaatcaga 901 gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa 961 agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg 1021 ccgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg 1081 ggcgggacgg cccttctcct ccgggctgta attagctgag caagaggtaa gggtttaagg 1141 gatggttggt tggtggggta ttaatgttta attacctgga gcacctgcct gaaatcactt 1201 tttttcaggt tggaccggtg ccaccatgta cccatacgat gttccagatt acgcttcgcc 1261 gaagaaaaag cgcaaggtcg aagcgtccga caagaagtac agcatcggcc tggccatcgg 1321 caccaactct gtgggctggg ccgtgatcac cgacgagtac aaggtgccca gcaagaaatt 1381 caaggtgctg ggcaacaccg accggcacag catcaagaag aacctgatcg gagccctgct 1441 gttcgacagc ggcgaaacag ccgaggccac ccggctgaag agaaccgcca gaagaagata 1501 caccagacgg aagaaccgga tctgctatct gcaagagatc ttcagcaacg agatggccaa 1561 ggtggacgac agcttcttcc acagactgga agagtccttc ctggtggaag aggataagaa 1621 gcacgagcgg caccccatct tcggcaacat cgtggacgag gtggcctacc acgagaagta 1681 ccccaccatc taccacctga gaaagaaact ggtggacagc accgacaagg ccgacctgcg 1741 gctgatctat ctggccctgg cccacatgat caagttccgg ggccacttcc tgatcgaggg 1801 cgacctgaac cccgacaaca gcgacgtgga caagctgttc atccagctgg tgcagaccta 1861 caaccagctg ttcgaggaaa accccatcaa cgccagcggc gtggacgcca aggccatcct 1921 gtctgccaga ctgagcaaga gcagacggct ggaaaatctg atcgcccagc tgcccggcga 1981 gaagaagaat ggcctgttcg gcaacctgat tgccctgagc ctgggcctga cccccaactt 2041 caagagcaac ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg acacctacga 2101 cgacgacctg gacaacctgc tggcccagat cggcgaccag tacgccgacc tgtttctggc 2161 cgccaagaac ctgtccgacg ccatcctgct gagcgacatc ctgagagtga acaccgagat 2221 caccaaggcc cccctgagcg cctctatgat caagagatac gacgagcacc accaggacct 2281 gaccctgctg aaagctctcg tgcggcagca gctgcctgag aagtacaaag agattttctt 2341 cgaccagagc aagaacggct acgccggcta cattgacggc ggagccagcc aggaagagtt 2401 ctacaagttc atcaagccca tcctggaaaa gatggacggc accgaggaac tgctcgtgaa 2461 gctgaacaga gaggacctgc tgcggaagca gcggaccttc gacaacggca gcatccccca 2521 ccagatccac ctgggagagc tgcacgccat tctgcggcgg caggaagatt tttacccatt 2581 cctgaaggac aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc cctactacgt 2641 gggccctctg gccaggggaa acagcagatt cgcctggatg accagaaaga gcgaggaaac 2701 catcaccccc tggaacttcg aggaagtggt ggacaagggc gcttccgccc agagcttcat 2761 cgagcggatg accaacttcg ataagaacct gcccaacgag aaggtgctgc ccaagcacag 2821 cctgctgtac gagtacttca ccgtgtataa cgagctgacc aaagtgaaat acgtgaccga 2881 gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg tggacctgct 2941 gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa gaggactact tcaagaaaat 3001 cgagtgcttc gactccgtgg aaatctccgg cgtggaagat cggttcaacg cctccctggg 3061 cacataccac gatctgctga aaattatcaa ggacaaggac ttcctggaca atgaggaaaa 3121 cgaggacatt ctggaagata tcgtgctgac cctgacactg tttgaggaca gagagatgat 3181 cgaggaacgg ctgaaaacct atgcccacct gttcgacgac aaagtgatga agcagctgaa 3241 gcggcggaga tacaccggct ggggcaggct gagccggaag ctgatcaacg gcatccggga 3301 caagcagtcc ggcaagacaa tcctggattt cctgaagtcc gacggcttcg ccaacagaaa 3361 cttcatgcag ctgatccacg acgacagcct gacctttaaa gaggacatcc agaaagccca 3421 ggtgtccggc cagggcgata gcctgcacga gcacattgcc aatctggccg gcagccccgc 3481 cattaagaag ggcatcctgc agacagtgaa ggtggtggac gagctcgtga aagtgatggg 3541 ccggcacaag cccgagaaca tcgtgatcga aatggccaga gagaaccaga ccacccagaa 3601 gggacagaag aacagccgcg agagaatgaa gcggatcgaa gagggcatca aagagctggg 3661 cagccagatc ctgaaagaac accccgtgga aaacacccag ctgcagaacg agaagctgta 3721 cctgtactac ctgcagaatg ggcgggatat gtacgtggac caggaactgg acatcaaccg 3781 gctgtccgac tacgatgtgg acgctatcgt gcctcagagc tttctgaagg acgactccat 3841 cgacaacaag gtgctgacca gaagcgacaa gaaccggggc aagagcgaca acgtgccctc 3901 cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg ccaagctgat 3961 tacccagaga aagttcgaca atctgaccaa ggccgagaga ggcggcctga gcgaactgga 4021 taaggccggc ttcatcaaga gacagctggt ggaaacccgg cagatcacaa agcacgtggc 4081 acagatcctg gactcccgga tgaacactaa gtacgacgag aatgacaagc tgatccggga 4141 agtgaaagtg atcaccctga agtccaagct ggtgtccgat ttccggaagg atttccagtt 4201 ttacaaagtg cgcgagatca acaactacca ccacgcccac gacgcctacc tgaacgccgt 4261 cgtgggaacc gccctgatca aaaagtaccc taagctggaa agcgagttcg tgtacggcga 4321 ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc 4381 taccgccaag tacttcttct acagcaacat catgaacttt ttcaagaccg agattaccct 4441 ggccaacggc gagatccgga agcggcctct gatcgagaca aacggcgaaa ccggggagat 4501 cgtgtgggat aagggccggg attttgccac cgtgcggaaa gtgctgagca tgccccaagt 4561 gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc agcaaagagt ctatcctgcc 4621 caagaggaac agcgataagc tgatcgccag aaagaaggac tgggacccta agaagtacgg 4681 cggcttcgac agccccaccg tggcctattc tgtgctggtg gtggccaaag tggaaaaggg 4741 caagtccaag aaactgaaga gtgtgaaaga gctgctgggg atcaccatca tggaaagaag 4801 cagcttcgag aagaatccca tcgactttct ggaagccaag ggctacaaag aagtgaaaaa 4861 ggacctgatc atcaagctgc ctaagtactc cctgttcgag ctggaaaacg gccggaagag 4921 aatgctggcc tctgccggcg aactgcagaa gggaaacgaa ctggccctgc cctccaaata 4981 tgtgaacttc ctgtacctgg ccagccacta tgagaagctg aagggctccc ccgaggataa 5041 tgagcagaaa cagctgtttg tggaacagca caagcactac ctggacgaga tcatcgagca 5101 gatcagcgag ttctccaaga gagtgatcct ggccgacgct aatctggaca aagtgctgtc 5161 cgcctacaac aagcaccggg ataagcccat cagagagcag gccgagaata tcatccacct 5221 gtttaccctg accaatctgg gagcccctgc cgccttcaag tactttgaca ccaccatcga 5281 ccggaagagg tacaccagca ccaaagaggt gctggacgcc accctgatcc accagagcat 5341 caccggcctg tacgagacac ggatcgacct gtctcagctg ggaggcgaca gccccaagaa 5401 gaagagaaag gtggaggcca gcggtggagg aagtggaggt ggttcaggtg gaggatccgg 5461 cgaagctgca ccctgtgact gtgatggagg tacacaaaaa gaaaaaggcc catattatac 5521 acaccttggg gcaggaccaa gtgtggctgc tgtcagggag ctcatggaga ctaggtttgg 5581 ccagaagggg aaggcaatcc ggattgagaa gatagtgttc acggggaagg aagggaagag 5641 ctctcagggc tgcccggtcg ccaagtgggt gatcagaaga agtggtcctg aagagaagct 5701 tatttgtttg gttcgtgagc gtgtagacca tcactgttcg acggctgtga tagttgtcct 5761 tatcctgctg tgggaaggta tccctcgcct gatggctgac cgcctgtaca aagagctcac 5821 tgagaacttg aggtcctaca gcggacatcc cacagaccga agatgtaccc tcaacaaaaa 5881 gcgtacctgc acctgtcaag gcatcgaccc aaaaacctgc ggagcgtcct tctcctttgg 5941 ctgttcgtgg agcatgtatt tcaacggctg taagtttggg aggagtgaaa accccagaaa 6001 attcagactt gctccaaact accccttaca taactactat aagagaatta ctggaatgag 6061 ttctgaagga agtgacgtga aaaccgggtg gatcattcca gaccgcaaga ccctcataag 6121 cagagaggaa aaacagcttg aaaagaattt acaagaattg gctacagtat tagctccact 6181 ttacaagcag atggctccag ttgcttatca aaatcaggtg gaatatgaag aagttgctgg 6241 agactgtcga cttggaaatg aagaggggcg tcctttctct ggtgtcacct gttgcatgga 6301 tttttgtgcc cattctcaca aggacattca caacatgcac aacggaagca ccgtggtgtg 6361 tacgttgatt cgagcagatg gccgtgacac aaattgtccc gaggatgaac aactccacgt 6421 cctgccacta taccggcttg cagacactga tgaatttggc tccgtggaag ggatgaaggc 6481 caaaatcaaa tctggggcca tccaagtcaa tgggccaacc aggaagaggc gactacgttt 6541 tactgagcct gttcctcgat gtgggaagag ggccaaaatg aagcagaacc acaataaatc 6601 aggttcacac aacactaaga gcttttcatc agcctcatct acttctcacc tagtgaaaga 6661 cgaatctaca gacttctgtc ccctgcaggc ttcctccgca gaaacatcta cctgtacgta 6721 cagtaaaaca gcctcaggtg ggtttgcaga aacaagtagt attctccact gcacaatgcc 6781 ttctggagca cacagtggtg ctaatgcagc tgctggggaa tgtactggaa cggtgcagcc 6841 tgccgaggtg gctgctcatc ctcaccagtc tcttcccaca gccgattctc ccgttcatgc 6901 tgagcctctc actagtccat ctgagcagct aacttctaac cagtcaaacc agcagctccc 6961 tctcctcagc aattctcaga aactggcttc ctgtcaggtg gaagatgagc ggcaccctga 7021 agcggatgag cctcagcacc ccgaggacga taacttgcct caacttgatg agttctggtc 7081 agacagtgag gagatctacg ccgatccttc ctttggtggc gtggcgatag cacccattca 7141 cggctcggtg ctcattgagt gcgctcggaa ggagcttcat gctaccacct ctttgcgctc 7201 ccccaaacga ggggtccctt ttcgtgtgtc ccttgtattc taccagcaca aaagcctaaa 7261 caagcctaat catggttttg atatcaacaa aattaagtgt aaatgcaaaa aagtaacgaa 7321 aaaaaagccc gcagaccggg agtgtcctga tgtatccccc gaagccaatt tatcacacca 7381 aattccttct cgagttgcat caaccttaac ccgagacaat gttgttaccg tgtccccata 7441 ctctctcact catgttgcgg gaccctacaa tcgttgggtc taagaattcc tagagctcgc 7501 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg 7561 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt 7621 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc 7681 aagggggagg attgggaaga gaatagcagg catgctgggg agcggccgca ggaaccccta 7741 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca 7801 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc 7861 tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac 7921 cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg 7981 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg 8041 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg 8101 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt 8161 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt 8221 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta 8281 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa 8341 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt 8401 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc 8461 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac 8521 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac 8581 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa 8641 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt 8701 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc 8761 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc 8821 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa 8881 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg 8941 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag 9001 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc 9061 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta 9121 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg 9181 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca 9241 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac 9301 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat 9361 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg 9421 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata 9481 aatctggagc cggtgagcgt ggaagccgcg gtatcattgc agcactgggg ccagatggta 9541 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa 9601 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag 9661 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg 9721 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact 9781 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg 9841 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc 9901 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata 9961 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta 10021 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc 10081 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg 10141 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac 10201 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg 10261 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt 10321 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct 10381 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg 10441 ccttttgctg gccttttgct cacatgt //