LOCUS Exported 13028 bp ds-DNA circular SYN 01-OCT-2021 DEFINITION Expression of dCas12f-VPR-mCherry in mammalian cells. ACCESSION . VERSION . KEYWORDS pSLQ7334: pHR-PGK-SV40_NLS-dCas12f-VPR-c- Myc_NLS-mCherry-WPRE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 13028) AUTHORS Xu X, Chemparathy A, Zeng L, Kempton HR, Shang S, Nakamura M, Qi LS TITLE Engineered miniature CRISPR-Cas system for mammalian genome regulation and editing. JOURNAL Mol Cell. 2021 Aug 26. pii: S1097-2765(21)00648-1. doi: 10.1016/j.molcel.2021.08.008. PUBMED 34480847 REFERENCE 2 (bases 1 to 13028) AUTHORS . TITLE Direct Submission JOURNAL Exported Oct 1, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..13028 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 27..144 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 209..708 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(244..267) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 641..660 /label=mPGK-F /note="Mouse PGK promoter, forward primer" regulatory 768..777 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 780..800 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 2400..2426 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 2478..2627 /codon_start=1 /product="tetrameric repeat of the minimal activation domain of herpes simplex virus VP16 (Beerli et al., 1998)" /label=VP64 /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF DLDML" CDS 3072..3443 /codon_start=1 /product="transcriptional activation domain of human RelA, also known as p65 (O'Shea and Perkins, 2008)" /label=RelA (p65) AD /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD EDFSSIADMDFSALLSQISS" CDS 3462..4031 /codon_start=1 /product="transcriptional activation domain from the human herpesvirus 4 (Epstein-Barr virus) replication and transcription activator Rta/BRLF1 (Hardwick et al., 1992; Chavez et al., 2015)" /label=Rta AD /translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL DTFLNDECLLHAMHISTGLSIFDTSLF" CDS 4059..4085 /codon_start=1 /product="nuclear localization signal of human c-Myc proto-oncogene (Dang and Lee, 1988)" /label=c-myc NLS /translation="PAAKRVKLD" CDS 4098..4805 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE GRHSTGGMDELYK" primer_bind complement(4240..4258) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 4508..4527 /label=mCherry-F /note="mCherry, forward primer" misc_feature 4869..5457 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(4922..4942) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5340..5351) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(5460..5476) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5461..5477) /label=pBluescriptKS /note="For pBluescript vector" LTR 5567..5800 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" promoter complement(5825..5843) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(5826..5843) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(6133..6152) /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind complement(6366..6385) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 6485..6507 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(6545..6563) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 6631..6735 /gene="bla" /label=AmpR promoter CDS 6736..7596 /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(6954..6973) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7767..8355 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8256..8275 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 8509..8526 /label=L4440 /note="L4440 vector, forward primer" primer_bind complement(8596..8616) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 8601..8930 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 8781..8916 /label=SV40 ori /note="SV40 origin of replication" primer_bind 8843..8862 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" intron 10064..10129 /label=small t intron /note="SV40 (simian virus 40) small t antigen intron" CDS 10259..10279 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" polyA_signal 10704..10838 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(10741..10760) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 10795..10814 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" LTR 11006..11639 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 11686..11811 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 12308..12541 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 12726..12770 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" ORIGIN 1 cgccaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt 61 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa 121 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccagtttgg 181 atcgataagc ttgatatcga attctaccgg gtaggggagg cgcttttccc aaggcagtct 241 ggagcatgcg ctttagcagc cccgctgggc acttggcgct acacaagtgg cctctggcct 301 cgcacacatt ccacatccac cggtaggcgc caaccggctc cgttctttgg tggccccttc 361 gcgccacctt ctactcctcc cctagtcagg aagttccccc ccgccccgca gctcgcgtcg 421 tgcaggacgt gacaaatgga agtagcacgt ctcactagtc tcgtgcagat ggacagcacc 481 gctgagcaat ggaagcgggt aggcctttgg ggcagcggcc aatagcagct ttgctccttc 541 gctttctggg ctcagaggct gggaaggggt gggtccgggg gcgggctcag gggcgggctc 601 aggggcgggg cgggcgcccg aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag 661 cgcacgtctg ccgcgctgtt ctcctcttcc tcatctccgg gcctttcgac ctgcagccca 721 agcttacgcg tctgcaggat atcaagcttg cggtaccgcg ggcccgggcc accatgggac 781 ccaagaaaaa acgcaaggtg ggaagcggat ccgccaaaaa caccattacc aaaacactga 841 aactgcgtat tgtgcgtccg tataatagcg cagaagtgga aaaaattgtt gccgacgaaa 901 aaaacaaccg cgaaaaaatc gcactggaaa agaacaaaga caaagtgaaa gaagcctgca 961 gcaaacatct gaaagttgca gcatattgta ccacacaggt tgaacgtaat gcatgcctgt 1021 tttgtaaagc acgtaaactg gatgacaaat tctaccaaaa actgcgtggt cagtttccgg 1081 atgcagtttt ttggcaagaa atcagcgaaa tttttcgcca gctgcagaaa caggcagcag 1141 aaatctataa tcagagcctg atcgaactgt actacgagat ttttatcaaa ggcaaaggta 1201 ttgcaaatgc cagcagcgtt gaacattatc tgagtgatgt ttgttatacc cgtgcagcag 1261 aactgtttaa aaacgcagca attgcaagcg gtctgcgtag caaaatcaaa agcaattttc 1321 gtctgaaaga actgaaaaac atgaaaagtg gtctgccgac caccaaaagc gataattttc 1381 cgattccgct ggttaaacag aaaggtggtc agtataccgg ttttgaaatt agcaatcata 1441 atagcgactt catcatcaag attccgtttg gtcgttggca ggtcaaaaaa gagattgata 1501 aatatcgtcc gtgggagaaa tttgactttg aacaggttca gaaaagcccg aaaccgatta 1561 gcctgctgct gagcacccag cgtcgtaaac gtaataaagg ttggagcaaa gatgaaggca 1621 ccgaagccga aatcaaaaaa gttatgaatg gcgattatca gaccagctac attgaagtta 1681 aacgtggcag caaaatctgt gaaaaaagcg catggatgct gaatctgagc attgatgttc 1741 cgaaaattga taaaggtgtg gatccgagca ttattggtgg tattgcagtt ggtgttaaat 1801 caccgctggt ttgcgcaatt aacaatgcat ttagccgtta tagcatcagc gataacgacc 1861 tgtttcactt caacaagaaa atgtttgcac gtcgtcgtat cctgctgaaa aaaaaccgtc 1921 ataaacgtgc aggtcatggt gcaaaaaaca aactgaaacc gatcaccatt ctgaccgaaa 1981 aaagtgaacg ttttcgcaaa aagctgattg aacgttgggc atgtgaaatc gcggatttct 2041 tcattaaaaa caaagttggc accgtgcaga tggaaaatct ggaaagcatg aaacgtaaag 2101 aggacagcta ttttaacatt cgcctgcgtg gcttttggcc gtatgcagaa atgcagaaca 2161 aaatcgaatt caaactgaag cagtatggca tcgaaattcg taaagttgca ccgaataata 2221 ccagcaaaac ctgtagcaaa tgtggccatc tgaacaacta tttcaacttc gagtaccgca 2281 agaaaaacaa attcccgcac tttaaatgcg aaaaatgcaa cttcaaagaa aacgccgcgt 2341 ataatgcagc cctgaatatt tcaaacccga aactgaaaag caccaaagag gaaccggcct 2401 atccctatga cgtgcccgat tatgccagcc tgggcagcgg cgacggcatt ggtagtggga 2461 gcaacggcag cagcctcgat gctttagacg attttgactt agatatgctt ggttcagacg 2521 cgttagacga cttcgaccta gacatgttag gctcagatgc attggacgac ttcgatttag 2581 atatgttggg ctccgatgcc ctagatgact ttgatctaga tatgctaggt agtggcggca 2641 gcggatccca gtatctgccc gacacagatg atagacaccg aatcgaagag aaacgcaagc 2701 gaacgtatga aaccttcaaa tcgatcatga agaaatcgcc cttctcgggt ccgaccgatc 2761 ccaggccccc accgagaagg attgcggtcc cgtcccgctc gtcggccagc gtgccgaagc 2821 ctgcgccgca gccctacccc ttcacgtcga gcctgagcac aatcaattat gacgagttcc 2881 cgacgatggt gttcccctcg ggacaaatct cacaagcctc ggcgctcgca ccagcgcctc 2941 cccaagtcct tccgcaagcg cctgccccag cgcctgcacc ggcaatggtg tccgccctcg 3001 cacaggcccc tgcgcccgtc cccgtgctcg cgcctggacc gccccaggcg gtcgctccac 3061 cggctccgaa gccgacgcag gccggagagg gaacactctc cgaagcactt cttcaactcc 3121 agtttgatga cgaggatctt ggagcactcc ttggaaactc gacagaccct gcggtgttta 3181 ccgacctcgc gtcagtagat aactccgaat ttcagcagct tttgaaccag ggtatcccgg 3241 tcgcgccaca tacaacggag cccatgttga tggaataccc cgaagcaatc acgagacttg 3301 tgacgggagc gcagcggcct cccgatcccg cacccgcacc tttgggggca cctggcctcc 3361 ctaacggact tttgagcggc gacgaggatt tctcctccat cgccgatatg gatttctcag 3421 ccttgctgtc acagatttcc agcggctctg gcagcggcag ccgggattcc agggaaggga 3481 tgtttttgcc gaagcctgag gccggctccg ctattagtga cgtgtttgag ggccgcgagg 3541 tgtgccagcc aaaacgaatc cggccatttc atcctccagg aagtccatgg gccaaccgcc 3601 cactccccgc cagcctcgca ccaacaccaa ccggtccagt acatgagcca gtcgggtcac 3661 tgaccccggc accagtccct cagccactgg atccagcgcc cgcagtgact cccgaggcca 3721 gtcacctgtt ggaggatccc gatgaagaga cgagccaggc tgtcaaagcc cttcgggaga 3781 tggccgatac tgtgattccc cagaaggaag aggctgcaat ctgtggccaa atggaccttt 3841 cccatccgcc cccaaggggc catctggatg agctgacaac cacacttgag tccatgaccg 3901 aggatctgaa cctggactca cccctgaccc cggaattgaa cgagattctg gataccttcc 3961 tgaacgacga gtgcctcttg catgccatgc atatcagcac aggactgtcc atcttcgaca 4021 catctctgtt tcatatgggt ggaggctccg gggaagatcc tgctgccaaa cgcgttaaac 4081 tagacatggg aagcggagtg agcaagggcg aggaggataa catggccatc atcaaggagt 4141 tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg 4201 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg 4261 gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg 4321 cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct 4381 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact 4441 cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct 4501 ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt 4561 accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg 4621 gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg 4681 gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg 4741 tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca 4801 agtaagcggc cgcataagac tctagagtcg acctgcaggc atgcaagctt gatatcaagc 4861 ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact 4921 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg 4981 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg 5041 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa 5101 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc 5161 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg 5221 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt 5281 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt 5341 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc 5401 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc 5461 gataccgtcg acctcgaggg aattaattcg agctcggtac ctttaagacc aatgacttac 5521 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt 5581 cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag 5641 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc 5701 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga 5761 tccctcagac ccttttagtc agtgtggaaa atctctagca gcatctagaa ttaattccgt 5821 gtattctata gtgtcaccta aatcgtatgt gtatgataca taaggttatg tattaattgt 5881 agccgcgttc taacgacaat atgtacaagc ctaattgtgt agcatctggc ttactgaagc 5941 agaccctatc atctctctcg taaactgccg tcagagtcgg tttggttgga cgaaccttct 6001 gagtttctgg taacgccgtc ccgcacccgg aaatggtcag cgaaccaatc agcagggtca 6061 tcgctagcca gatcctctac gccggacgca tcgtggccgg catcaccggc gccacaggtg 6121 cggttgctgg cgcctatatc gccgacatca ccgatgggga agatcgggct cgccacttcg 6181 ggctcatgag cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc gggggactgt 6241 tgggcgccat ctccttgcat gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc 6301 tactactggg ctgcttccta atgcaggagt cgcataaggg agagcgtcga atggtgcact 6361 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc 6421 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc 6481 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga 6541 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag 6601 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa 6661 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat 6721 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg 6781 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa 6841 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt 6901 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt 6961 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat 7021 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg 7081 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta 7141 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat 7201 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag 7261 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa 7321 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca 7381 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc 7441 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt 7501 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc 7561 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat 7621 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt 7681 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac 7741 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc 7801 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca 7861 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta 7921 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct 7981 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg 8041 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc 8101 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta 8161 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg 8221 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt 8281 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg 8341 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg 8401 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc 8461 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg 8521 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt 8581 cattaatgca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag 8641 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc 8701 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag 8761 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc 8821 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc 8881 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagcttggac 8941 acaagacagg cttgcgagat atgtttgaga ataccacttt atcccgcgtc agggagaggc 9001 agtgcgtaaa aagacgcgga ctcatgtgaa atactggttt ttagtgcgcc agatctctat 9061 aatctcgcgc aacctatttt cccctcgaac actttttaag ccgtagataa acaggctggg 9121 acacttcaca tgagcgaaaa atacatcgtc acctgggaca tgttgcagat ccatgcacgt 9181 aaactcgcaa gccgactgat gccttctgaa caatggaaag gcattattgc cgtaagccgt 9241 ggcggtctgt accgggtgcg ttactggcgc gtgaactggg tattcgtcat gtcgataccg 9301 tttgtatttc cagctacgat cacgacaacc agcgcgagct taaagtgctg aaacgcgcag 9361 aaggcgatgg cgaaggcttc atcgttattg atgacctggt ggataccggt ggtactgcgg 9421 ttgcgattcg tgaaatgtat ccaaaagcgc actttgtcac catcttcgca aaaccggctg 9481 gtcgtccgct ggttgatgac tatgttgttg atatcccgca agatacctgg attgaacagc 9541 cgtgggatat gggcgtcgta ttcgtcccgc caatctccgg tcgctaatct tttcaacgcc 9601 tggcactgcc gggcgttgtt ctttttaact tcaggcgggt tacaatagtt tccagtaagt 9661 attctggagg ctgcatccat gacacaggca aacctgagcg aaaccctgtt caaaccccgc 9721 tttaaacatc ctgaaacctc gacgctagtc cgccgcttta atcacggcgc acaaccgcct 9781 gtgcagtcgg cccttgatgg taaaaccatc cctcactggt atcgcatgat taaccgtctg 9841 atgtggatct ggcgcggcat tgacccacgc gaaatcctcg acgtccaggc acgtattgtg 9901 atgagcgatg ccgaacgtac cgacgatgat ttatacgata cggtgattgg ctaccgtggc 9961 ggcaactgga tttatgagtg ggccccggat ctttgtgaag gaaccttact tctgtggtgt 10021 gacataattg gacaaactac ctacagagat ttaaagctct aaggtaaata taaaattttt 10081 aagtgtataa tgtgttaaac tactgattct aattgtttgt gtattttaga ttccaaccta 10141 tggaactgat gaatgggagc agtggtggaa tgcctttaat gaggaaaacc tgttttgctc 10201 agaagaaatg ccatctagtg atgatgaggc tactgctgac tctcaacatt ctactcctcc 10261 aaaaaagaag agaaaggtag aagaccccaa ggactttcct tcagaattgc taagtttttt 10321 gagtcatgct gtgtttagta atagaactct tgcttgcttt gctatttaca ccacaaagga 10381 aaaagctgca ctgctataca agaaaattat ggaaaaatat tctgtaacct ttataagtag 10441 gcataacagt tataatcata acatactgtt ttttcttact ccacacaggc atagagtgtc 10501 tgctattaat aactatgctc aaaaattgtg tacctttagc tttttaattt gtaaaggggt 10561 taataaggaa tatttgatgt atagtgcctt gactagagat cataatcagc cataccacat 10621 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata 10681 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa 10741 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt 10801 tgtccaaact catcaatgta tcttatcatg tctggatcaa ctggataact caagctaacc 10861 aaaatcatcc caaacttccc accccatacc ctattaccac tgccaattac ctgtggtttc 10921 atttactcta aacctgtgat tcctctgaat tattttcatt ttaaagaaat tgtatttgtt 10981 aaatatgtac tacaaactta gtagttggaa gggctaattc actcccaaag aagacaagat 11041 atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattagca gaactacaca 11101 ccagggccag gggtcagata tccactgacc tttggatggt gctacaagct agtaccagtt 11161 gagccagata aggtagaaga ggccaataaa ggagagaaca ccagcttgtt acaccctgtg 11221 agcctgcatg ggatggatga cccggagaga gaagtgttag agtggaggtt tgacagccgc 11281 ctagcatttc atcacgtggc ccgagagctg catccggagt acttcaagaa ctgctgatat 11341 cgagcttgct acaagggact ttccgctggg gactttccag ggaggcgtgg cctgggcggg 11401 actggggagt ggcgagccct cagatcctgc atataagcag ctgctttttg cctgtactgg 11461 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact 11521 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg 11581 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag 11641 tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg 11701 actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca 11761 aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag 11821 cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa 11881 tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct 11941 ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt 12001 cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg 12061 catcaaagga tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa 12121 aacaaaagta agaccaccgc acagcaagcg gccggccgct gatcttcaga cctggaggag 12181 gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac 12241 cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag 12301 tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag 12361 cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga 12421 acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca 12481 tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc 12541 tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta 12601 gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca 12661 gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc 12721 aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt 12781 ttaacataac aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg 12841 taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt 12901 caccattatc gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa 12961 tagaagaaga aggtggagag agagacagag acagatccat tcgattagtg aacggatctc 13021 gacggtat //