LOCUS Exported 9320 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Endogenous tagging of Tarp-gamma-8: Intramolecular (amino acid position: A408). ACCESSION . VERSION . KEYWORDS pORANGE CACNG8-GFP KI #2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9320) 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 9320) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9320 /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 559..1269 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELY" CDS 559..1269 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELY" primer_bind complement(601..622) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(862..881) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1209..1230 /label=EGFP-C /note="EGFP, forward primer" enhancer 1385..1670 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer; contains an 18-bp deletion relative to the standard CMV enhancer" promoter 1672..1949 /label=chicken beta-actin promoter intron 1950..2177 /label=hybrid intron /note="hybrid between chicken beta-actin (CBA) and minute virus of mice (MMV) introns (Gray et al., 2011)" CDS 2198..2224 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 2228..2248 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 2258..6358 /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 6362..6382 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" primer_bind complement(6422..6439) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 6428..6635 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" repeat_region 6644..6784 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 6644..6773 /label=AAV2 ITR rep_origin 6859..7314 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6946..6965) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 7156..7177 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(7331..7350) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 7450..7472 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(7510..7528) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 7596..7700 /gene="bla" /label=AmpR promoter CDS 7701..8561 /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(7919..7938) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 8732..9320 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 9221..9240 /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 gcgtgttggt gttggacgcg ggttttagag ctagaaatag caagttaaaa 361 taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt tttgttttag 421 agctagaaat agcaagttaa aataaggcta gtccgttttt agcgcgtgcg ccaattctgc 481 agacaaatgg ctctagaagc ttgcgtgttg gtgttggacg cggcggctag cgcaggtagc 541 ggctccagta gatctggggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc 601 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag 661 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc 721 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac 781 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag 841 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc 901 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc 961 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc 1021 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc 1081 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg 1141 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag 1201 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac 1261 gagctgtact ctaggggagc tggcgctgga gcgggtgcag gggctggcag cgctggcgtg 1321 ttggtgttgg acgcggcgga ctcgaggatc ctaggacgcg tcctaggacg cgtccagagg 1381 tacccgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc 1441 cattgacgtc aatagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt 1501 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta 1561 ttgacgtcaa tgacggtaaa tggcccgcct ggcattgtgc ccagtacatg accttatggg 1621 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtcgaggtga 1681 gccccacgtt ctgcttcact ctccccatct cccccccctc cccaccccca attttgtatt 1741 tatttatttt ttaattattt tgtgcagcga tgggggcggg gggggggggg gggcgcgcgc 1801 caggcggggc ggggcggggc gaggggcggg gcggggcgag gcggagaggt gcggcggcag 1861 ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg cggcggcggc 1921 cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt cgccccgtgc 1981 cccgctccgc cgccgcctcg cgccgcccgc cccggctctg actgaccgcg ttactcccac 2041 aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa ttagctgagc aagaggtaag 2101 ggtttaaggg atggttggtt ggtggggtat taatgtttaa ttacctggag cacctgcctg 2161 aaatcacttt ttttcaggtt ggaccggtgc caccatgtac ccatacgatg ttccagatta 2221 cgcttcgccg aagaaaaagc gcaaggtcga agcgtccgac aagaagtaca gcatcggcct 2281 ggacatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag 2341 caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg 2401 agccctgctg ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga gaaccgccag 2461 aagaagatac accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga 2521 gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga 2581 ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca 2641 cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc 2701 cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct 2761 gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt 2821 gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa 2881 ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga tcgcccagct 2941 gcccggcgag aagaagaatg gcctgttcgg caacctgatt gccctgagcc tgggcctgac 3001 ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc tgagcaagga 3061 cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt acgccgacct 3121 gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc tgagagtgaa 3181 caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg acgagcacca 3241 ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga agtacaaaga 3301 gattttcttc gaccagagca agaacggcta cgccggctac attgacggcg gagccagcca 3361 ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca ccgaggaact 3421 gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg acaacggcag 3481 catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc aggaagattt 3541 ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct tccgcatccc 3601 ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga ccagaaagag 3661 cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg cttccgccca 3721 gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc 3781 caagcacagc ctgctgtacg agtacttcac cgtgtataac gagctgacca aagtgaaata 3841 cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa aggccatcgt 3901 ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag aggactactt 3961 caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc ggttcaacgc 4021 ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact tcctggacaa 4081 tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag 4141 agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa 4201 gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg 4261 catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc 4321 caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca 4381 gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca atctggccgg 4441 cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg agctcgtgaa 4501 agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag agaaccagac 4561 cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag agggcatcaa 4621 agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc tgcagaacga 4681 gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc aggaactgga 4741 catcaaccgg ctgtccgact acgatgtgga ccatatcgtg cctcagagct ttctgaagga 4801 cgactccatc gacaacaagg tgctgaccag aagcgacaag aaccggggca agagcgacaa 4861 cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcggcagc tgctgaacgc 4921 caagctgatt acccagagaa agttcgacaa tctgaccaag gccgagagag gcggcctgag 4981 cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc agatcacaaa 5041 gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga atgacaagct 5101 gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccggaagga 5161 tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg acgcctacct 5221 gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt 5281 gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg agcaggaaat 5341 cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt tcaagaccga 5401 gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa acggcgaaac 5461 cggggagatc gtgtgggata agggccggga ttttgccacc gtgcggaaag tgctgagcat 5521 gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc 5581 tatcctgccc aagaggaaca gcgataagct gatcgccaga aagaaggact gggaccctaa 5641 gaagtacggc ggcttcgaca gccccaccgt ggcctattct gtgctggtgg tggccaaagt 5701 ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctgctgggga tcaccatcat 5761 ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg gctacaaaga 5821 agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg 5881 ccggaagaga atgctggcct ctgccggcga actgcagaag ggaaacgaac tggccctgcc 5941 ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga agggctcccc 6001 cgaggataat gagcagaaac agctgtttgt ggaacagcac aagcactacc tggacgagat 6061 catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta atctggacaa 6121 agtgctgtcc gcctacaaca agcaccggga taagcccatc agagagcagg ccgagaatat 6181 catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt actttgacac 6241 caccatcgac cggaagaggt acaccagcac caaagaggtg ctggacgcca ccctgatcca 6301 ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg gaggcgacag 6361 ccccaagaag aagagaaagg tggaggccag ctaagaattc taactagagc tcgctgatca 6421 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc 6481 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg 6541 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg 6601 gaggattggg aagagaatag caggcatgct ggggagcggc cgcaggaacc cctagtgatg 6661 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc 6721 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg 6781 caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata 6841 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg 6901 ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct 6961 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc 7021 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg 7081 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt 7141 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg 7201 gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc 7261 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt 7321 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa 7381 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg 7441 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga 7501 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt 7561 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt 7621 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat 7681 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt 7741 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg 7801 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga 7861 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc 7921 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac 7981 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg 8041 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca 8101 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg 8161 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg 8221 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg 8281 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag 8341 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg 8401 gagccggtga gcgtggaagc cgcggtatca ttgcagcact ggggccagat ggtaagccct 8461 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac 8521 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact 8581 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga 8641 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt 8701 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct 8761 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc 8821 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc 8881 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc 8941 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg 9001 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt 9061 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg 9121 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg 9181 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt 9241 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag 9301 gggggcggag cctatggaaa //