LOCUS Exported 9270 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Endogenous tagging of C9orf4: C-terminal (amino acid position: STOP codon). ACCESSION . VERSION . KEYWORDS pORANGE C9orf4-GFP KI SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9270) AUTHORS Willems J, de Jong APH, Scheefhals N, Mertens E, Catsburg LAE, Poorthuis RB, de Winter F, Verhaagen J, Meye FJ, MacGillavry HD TITLE ORANGE: A CRISPR/Cas9-based genome editing toolbox for epitope tagging of endogenous proteins in neurons. JOURNAL PLoS Biol. 2020 Apr 10;18(4):e3000665. doi: 10.1371/journal.pbio.3000665. PUBMED 32275651 REFERENCE 2 (bases 1 to 9270) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9270 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 62..302 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 62..82 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 233..252 /label=LKO.1 5' /note="Human U6 promoter, forward primer" misc_RNA 332..407 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" CDS 551..1267 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" CDS 551..1264 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(593..614) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(854..873) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1201..1222 /label=EGFP-C /note="EGFP, forward primer" enhancer 1335..1620 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer; contains an 18-bp deletion relative to the standard CMV enhancer" promoter 1622..1899 /label=chicken beta-actin promoter intron 1900..2127 /label=hybrid intron /note="hybrid between chicken beta-actin (CBA) and minute virus of mice (MMV) introns (Gray et al., 2011)" CDS 2148..2174 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 2178..2198 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 2208..6308 /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD ATLIHQSITGLYETRIDLSQLGGD" CDS 6312..6332 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" primer_bind complement(6372..6389) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 6378..6585 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" repeat_region 6594..6734 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 6594..6723 /label=AAV2 ITR rep_origin 6809..7264 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6896..6915) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 7106..7127 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(7281..7300) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 7400..7422 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(7460..7478) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 7546..7650 /gene="bla" /label=AmpR promoter CDS 7651..8511 /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(7869..7888) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 8682..9270 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 9171..9190 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg 61 tgagggccta tttcccatga ttccttcata tttgcatata cgatacaagg ctgttagaga 121 gataattgga attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag 181 aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca 241 tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg 301 acgaaacacc gtggcttgca gcggaggtta agttttagag ctagaaatag caagttaaaa 361 taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt tttgttttag 421 agctagaaat agcaagttaa aataaggcta gtccgttttt agcgcgtgcg ccaattctgc 481 agacaaatgg ctctagaagc tttggcttgc agcggaggtt aagggaggta gcggctccag 541 tagatctggg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga 601 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc 661 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg 721 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca 781 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac 841 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga 901 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct 961 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca 1021 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca 1081 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga 1141 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca 1201 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta 1261 caagtaatgg cttgcagcgg aggttaaggg ctcgaggatc ctaggacgcg tcctaggacg 1321 cgtccagagg tacccgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac 1381 gacccccgcc cattgacgtc aatagtaacg ccaataggga ctttccattg acgtcaatgg 1441 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt 1501 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattgtgc ccagtacatg 1561 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg 1621 gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc cccaccccca 1681 attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg gggggggggg 1741 gggcgcgcgc caggcggggc ggggcggggc gaggggcggg gcggggcgag gcggagaggt 1801 gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg 1861 cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt 1921 cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc cccggctctg actgaccgcg 1981 ttactcccac aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa ttagctgagc 2041 aagaggtaag ggtttaaggg atggttggtt ggtggggtat taatgtttaa ttacctggag 2101 cacctgcctg aaatcacttt ttttcaggtt ggaccggtgc caccatgtac ccatacgatg 2161 ttccagatta cgcttcgccg aagaaaaagc gcaaggtcga agcgtccgac aagaagtaca 2221 gcatcggcct ggacatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca 2281 aggtgcccag caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga 2341 acctgatcgg agccctgctg ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga 2401 gaaccgccag aagaagatac accagacgga agaaccggat ctgctatctg caagagatct 2461 tcagcaacga gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc 2521 tggtggaaga ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg 2581 tggcctacca cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca 2641 ccgacaaggc cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg 2701 gccacttcct gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca 2761 tccagctggt gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg 2821 tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga 2881 tcgcccagct gcccggcgag aagaagaatg gcctgttcgg caacctgatt gccctgagcc 2941 tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc 3001 tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt 3061 acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc 3121 tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg 3181 acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga 3241 agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac attgacggcg 3301 gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca 3361 ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg 3421 acaacggcag catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc 3481 aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct 3541 tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga 3601 ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg 3661 cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga 3721 aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac gagctgacca 3781 aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa 3841 aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag 3901 aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc 3961 ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact 4021 tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt 4081 ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca 4141 aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc 4201 tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg 4261 acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag 4321 aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca 4381 atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg 4441 agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag 4501 agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag 4561 agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc 4621 tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc 4681 aggaactgga catcaaccgg ctgtccgact acgatgtgga ccatatcgtg cctcagagct 4741 ttctgaagga cgactccatc gacaacaagg tgctgaccag aagcgacaag aaccggggca 4801 agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcggcagc 4861 tgctgaacgc caagctgatt acccagagaa agttcgacaa tctgaccaag gccgagagag 4921 gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc 4981 agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga 5041 atgacaagct gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt 5101 tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg 5161 acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa 5221 gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg 5281 agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt 5341 tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa 5401 acggcgaaac cggggagatc gtgtgggata agggccggga ttttgccacc gtgcggaaag 5461 tgctgagcat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca 5521 gcaaagagtc tatcctgccc aagaggaaca gcgataagct gatcgccaga aagaaggact 5581 gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct gtgctggtgg 5641 tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctgctgggga 5701 tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg 5761 gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc 5821 tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag ggaaacgaac 5881 tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga 5941 agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac aagcactacc 6001 tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta 6061 atctggacaa agtgctgtcc gcctacaaca agcaccggga taagcccatc agagagcagg 6121 ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt 6181 actttgacac caccatcgac cggaagaggt acaccagcac caaagaggtg ctggacgcca 6241 ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg 6301 gaggcgacag ccccaagaag aagagaaagg tggaggccag ctaagaattc taactagagc 6361 tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc 6421 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga 6481 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga 6541 cagcaagggg gaggattggg aagagaatag caggcatgct ggggagcggc cgcaggaacc 6601 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg 6661 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg 6721 cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc 6781 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg 6841 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct 6901 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat 6961 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt 7021 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg 7081 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac 7141 tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta 7201 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca 7261 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga 7321 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 7381 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 7441 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata 7501 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt 7561 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa 7621 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt 7681 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa 7741 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac 7801 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt 7861 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt 7921 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat 7981 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac 8041 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg 8101 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc 8161 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa 8221 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag 8281 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct 8341 gataaatctg gagccggtga gcgtggaagc cgcggtatca ttgcagcact ggggccagat 8401 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa 8461 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac 8521 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc 8581 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc 8641 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg 8701 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg 8761 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca 8821 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg 8881 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg 8941 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga 9001 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac 9061 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat 9121 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc 9181 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga 9241 tgctcgtcag gggggcggag cctatggaaa //