LOCUS Exported 14207 bp ds-DNA circular SYN 02-DEC-2021 DEFINITION Expresses ABA-inducible VPR-Sa dCas9. ACCESSION . VERSION . KEYWORDS pSLQ2833 pPB: CAG-PYL1-VPR-IRES-Zeo-WPRE-SV40PA PGK-ABI-tagBFP... SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 14207) AUTHORS Gao Y, Xiong X, Wong S, Charles EJ, Lim WA, Qi LS TITLE Complex transcriptional modulation with orthogonal and inducible dCas9 regulators. JOURNAL Nat Methods. 2016 Oct 24. doi: 10.1038/nmeth.4042. PUBMED 27776111 REFERENCE 2 (bases 1 to 14207) AUTHORS . TITLE Direct Submission JOURNAL Exported Dec 2, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..14207 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 7..284 /label=chicken beta-actin promoter intron 286..1303 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1227..1249 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1311..1330 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" regulatory 1358..1367 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1940..2089 /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 2096..2116 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 2546..2917 /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 2936..3505 /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" misc_feature 3523..4108 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(3690..3707) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 3917..3936 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 4124..4498 /codon_start=1 /gene="Sh ble from Streptoalloteichus hindustanus" /product="antibiotic-binding protein" /label=BleoR /note="confers resistance to bleomycin, phleomycin, and Zeocin(TM)" /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR EFALRDPAGNCVHFVAEEQD" misc_feature 4536..5124 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(4589..4609) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5007..5018) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(5163..5182) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5217..5236 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" promoter 5278..5777 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(5313..5336) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 5710..5729 /label=mPGK-F /note="Mouse PGK promoter, forward primer" CDS 6737..7435 /codon_start=1 /product="monomeric blue fluorescent protein" /label=TagBFP /note="mammalian codon-optimized" /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC DLPSKLGHKLN" CDS 7457..7477 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 10658..10705 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" repeat_region 10895..10957 /label=piggyBac right (3') inverted repeat /note="piggyBac transposon-specific inverted terminal repeat sequence (ITR)" primer_bind complement(11008..11028) /label=T3 /note="T3 promoter, forward primer" promoter complement(11008..11026) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(11047..11063) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(11047..11063) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(11060..11082) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 11071..11087 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(11095..11125) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 11140..11161 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(11278..11295) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(11449..12037) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(11529..11548) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(12208..13068) /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 12831..12850 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(13069..13173) /gene="bla" /label=AmpR promoter rep_origin complement(13199..13654) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(13336..13357) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 13548..13567 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 13781..13803 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 13795..13812 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 13796..13812 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 13822..13841 /label=T7 /note="T7 promoter, forward primer" promoter 13822..13840 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" repeat_region 13870..13904 /label=piggyBac left (5') inverted repeat /note="piggyBac transposon-specific inverted terminal repeat sequence (ITR)" ORIGIN 1 ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac 61 ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg 121 ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga 181 gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc 241 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcggggagt cgctgcgacg 301 ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact 361 gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta 421 gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct 481 ccgggagggc cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt 541 ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc 601 ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg 661 gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg tgggggggtg 721 agcagggggt gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc ctccccgagt 781 tgctgagcac ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg cggggctcgc 841 cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc cgcctcgggc 901 cggggagggc tcgggggagg ggcgcggcgg cccccggagc gccggcggct gtcgaggcgc 961 ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg gacttccttt 1021 gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct ctagcgggcg 1081 cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc 1141 gccgcgccgc cgtccccttc tccctctcca gcctcggggc tgtccgcggg gggacggctg 1201 ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg gcggctctag 1261 agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct 1321 ggttattgtg ctgtctcatc attttggcaa agaattcgcc accatgggtg ggggcgcgcc 1381 aactcaagac gaattcaccc aactctccca atcaatcgcc gagttccaca cgtaccaact 1441 cggtaacggc cgttgctcat ctctcctagc tcagcgaatc cacgcgccgc cggaaacagt 1501 atggtccgtg gtgagacgtt tcgataggcc acagatttac aaacacttca tcaaaagctg 1561 taacgtgagt gaagatttcg agatgcgagt gggatgcacg cgcgacgtga acgtgataag 1621 tggattaccg gcgaatacgt ctcgggagag attagatctg ttggacgatg atcggagagt 1681 gactgggttt agtataaccg gtggtgaaca taggctgagg aattataaat cggttacgac 1741 ggttcataga tttgagaaag aagaagaaga agaaaggatc tggaccgttg ttttggaatc 1801 ttatgttgtt gatgtaccgg aaggtaattc ggaggaagat acgagattgt ttgctgatac 1861 ggttattaga ttgaatcttc agaaacttgc ttcgatcact gaagctatga acgaattctc 1921 aagcgctgca ggtactagtg atgctttaga cgattttgac ttagatatgc ttggttcaga 1981 cgcgttagac gacttcgacc tagacatgtt aggctcagat gcattggacg acttcgattt 2041 agatatgttg ggctccgatg ccctagatga ctttgatcta gatatgctag gtagtcccaa 2101 aaagaagagg aaagtgggat cccagtatct gcccgacaca gatgatagac accgaatcga 2161 agagaaacgc aagcgaacgt atgaaacctt caaatcgatc atgaagaaat cgcccttctc 2221 gggtccgacc gatcccaggc ccccaccgag aaggattgcg gtcccgtccc gctcgtcggc 2281 cagcgtgccg aagcctgcgc cgcagcccta ccccttcacg tcgagcctga gcacaatcaa 2341 ttatgacgag ttcccgacga tggtgttccc ctcgggacaa atctcacaag cctcggcgct 2401 cgcaccagcg cctccccaag tccttccgca agcgcctgcc ccagcgcctg caccggcaat 2461 ggtgtccgcc ctcgcacagg cccctgcgcc cgtccccgtg ctcgcgcctg gaccgcccca 2521 ggcggtcgct ccaccggctc cgaagccgac gcaggccgga gagggaacac tctccgaagc 2581 acttcttcaa ctccagtttg atgacgagga tcttggagca ctccttggaa actcgacaga 2641 ccctgcggtg tttaccgacc tcgcgtcagt agataactcc gaatttcagc agcttttgaa 2701 ccagggtatc ccggtcgcgc cacatacaac ggagcccatg ttgatggaat accccgaagc 2761 aatcacgaga cttgtgacgg gagcgcagcg gcctcccgat cccgcacccg cacctttggg 2821 ggcacctggc ctccctaacg gacttttgag cggcgacgag gatttctcct ccatcgccga 2881 tatggatttc tcagccttgc tgtcacagat ttccagcggc tctggcagcg gcagccggga 2941 ttccagggaa gggatgtttt tgccgaagcc tgaggccggc tccgctatta gtgacgtgtt 3001 tgagggccgc gaggtgtgcc agccaaaacg aatccggcca tttcatcctc caggaagtcc 3061 atgggccaac cgcccactcc ccgccagcct cgcaccaaca ccaaccggtc cagtacatga 3121 gccagtcggg tcactgaccc cggcaccagt ccctcagcca ctggatccag cgcccgcagt 3181 gactcccgag gccagtcacc tgttggagga tcccgatgaa gagacgagcc aggctgtcaa 3241 agcccttcgg gagatggccg atactgtgat tccccagaag gaagaggctg caatctgtgg 3301 ccaaatggac ctttcccatc cgcccccaag gggccatctg gatgagctga caaccacact 3361 tgagtccatg accgaggatc tgaacctgga ctcacccctg accccggaat tgaacgagat 3421 tctggatacc ttcctgaacg acgagtgcct cttgcatgcc atgcatatca gcacaggact 3481 gtccatcttc gacacatctc tgttttgaat ttaaataatt cgcccctctc cctccccccc 3541 ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt 3601 attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt 3661 cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa 3721 tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac 3781 cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg 3841 tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt 3901 tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca 3961 gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacgtgtgt 4021 ttagtcgagg ttaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga 4081 aaaacacgat gataatatgg ccacaaccgg atccggagct accatggcca agttgaccag 4141 tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg 4201 gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt 4261 gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt 4321 gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt 4381 ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt 4441 cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgagt 4501 ttaaaccatg caagcttgat atcaagctta tcgataatca acctctggat tacaaaattt 4561 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg 4621 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt 4681 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg 4741 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc 4801 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg 4861 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt 4921 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc 4981 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg 5041 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga 5101 tctccctttg ggccgcctcc ccgcatcctt gtttattgca gcttataatg gttacaaata 5161 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg 5221 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc ctgcaggtaa ttctaccggg 5281 taggggaggc gcttttccca aggcagtctg gagcatgcgc tttagcagcc ccgctgggca 5341 cttggcgcta cacaagtggc ctctggcctc gcacacattc cacatccacc ggtaggcgcc 5401 aaccggctcc gttctttggt ggccccttcg cgccaccttc tactcctccc ctagtcagga 5461 agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc 5521 tcactaggct cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta ggcctttggg 5581 gcagcggcca atagcagctt tgctccttcg ctttctgggc tcagaggctg ggaaggggtg 5641 ggtccggggg cgggctcagg ggcgggctca ggggcggggc gggcgcccga aggtcctccg 5701 gaggcccggc attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct 5761 catctccggg cctttcgacc tgcagcccaa gcttgtcgac gccaccatga cgcgtgtgcc 5821 tttgtatggt tttacttcga tttgtggaag aagacctgag atggaagctg ctgtttcgac 5881 tataccaaga ttccttcaat cttcctctgg ttcgatgtta gatggtcggt ttgatcctca 5941 atccgccgct catttcttcg gtgtttacga cggccatggc ggttctcagg tagcgaacta 6001 ttgtagagag aggatgcatt tggctttggc ggaggagata gctaaggaga aaccgatgct 6061 ctgcgatggt gatacgtggc tggagaagtg gaagaaagct cttttcaact cgttcctgag 6121 agttgactcg gagattgagt cagttgcgcc ggagacggtt gggtcaacgt cggtggttgc 6181 cgttgttttc ccgtctcaca tcttcgtcgc taactgcggt gactctagag ccgttctttg 6241 ccgcggcaaa actgcacttc cattatccgt tgaccataaa ccggatagag aagatgaagc 6301 tgcgaggatt gaagccgcag gagggaaagt gattcagtgg aatggagctc gtgttttcgg 6361 tgttctcgcc atgtcgagat ccattggcga tagatacttg aaaccatcca tcattcctga 6421 tccggaagtg acggctgtga agagagtaaa agaagatgat tgtctgattt tggcgagtga 6481 cggggtttgg gatgtaatga cggatgaaga agcgtgtgag atggcaagga agcggattct 6541 cttgtggcac aagaaaaacg cggtggctgg ggatgcatcg ttgctcgcgg atgagcggag 6601 aaaggaaggg aaagatcctg cggcgatgtc cgcggctgag tatttgtcaa agctggcgat 6661 acagagagga agcaaagaca acataagtgt ggtggtggtt gatttgaagc atatgtcaag 6721 cgctgcaggt gctaccatga gcgagctgat taaggagaac atgcacatga agctgtacat 6781 ggagggcacc gtggacaacc atcacttcaa gtgcacatcc gagggcgaag gcaagcccta 6841 cgagggcacc cagaccatga gaatcaaggt ggtcgagggc ggccctctcc ccttcgcctt 6901 cgacatcctg gctactagct tcctctacgg cagcaagacc ttcatcaacc acacccaggg 6961 catccccgac ttcttcaagc agtccttccc tgagggcttc acatgggaga gagtcaccac 7021 atacgaagac gggggcgtgc tgaccgctac ccaggacacc agcctccagg acggctgcct 7081 catctacaac gtcaagatca gaggggtgaa cttcacatcc aacggccctg tgatgcagaa 7141 gaaaacactc ggctgggagg ccttcaccga gacgctgtac cccgctgacg gcggcctgga 7201 aggcagaaac gacatggccc tgaagctcgt gggcgggagc catctgatcg caaacatcaa 7261 gaccacatat agatccaaga aacccgctaa gaacctcaag atgcctggcg tctactatgt 7321 ggactacaga ctggaaagaa tcaaggaggc caacaacgag acctacgtcg agcagcacga 7381 ggtggcagtg gccagatact gcgacctccc tagcaaactg gggcacaagc ttaatggtgg 7441 aggctctgga atggccccaa agaagaagcg gaaggtcggt atccacggag tcccagcagc 7501 caagcggaac tacatcctgg gcctggccat cggcatcacc agcgtgggct acggcatcat 7561 cgactacgag acacgggacg tgatcgatgc cggcgtgcgg ctgttcaaag aggccaacgt 7621 ggaaaacaac gagggcaggc ggagcaagag aggcgccaga aggctgaagc ggcggaggcg 7681 gcatagaatc cagagagtga agaagctgct gttcgactac aacctgctga ccgaccacag 7741 cgagctgagc ggcatcaacc cctacgaggc cagagtgaag ggcctgagcc agaagctgag 7801 cgaggaagag ttctctgccg ccctgctgca cctggccaag agaagaggcg tgcacaacgt 7861 gaacgaggtg gaagaggaca ccggcaacga gctgtccacc aaagagcaga tcagccggaa 7921 cagcaaggcc ctggaagaga aatacgtggc cgaactgcag ctggaacggc tgaagaaaga 7981 cggcgaagtg cggggcagca tcaacagatt caagaccagc gactacgtga aagaagccaa 8041 acagctgctg aaggtgcaga aggcctacca ccagctggac cagagcttca tcgacaccta 8101 catcgacctg ctggaaaccc ggcggaccta ctatgaggga cctggcgagg gcagcccctt 8161 cggctggaag gacatcaaag aatggtacga gatgctgatg ggccactgca cctacttccc 8221 cgaggaactg cggagcgtga agtacgccta caacgccgac ctgtacaacg ccctgaacga 8281 cctgaacaat ctcgtgatca ccagggacga gaacgagaag ctggaatatt acgagaagtt 8341 ccagatcatc gagaacgtgt tcaagcagaa gaagaagccc accctgaagc agatcgccaa 8401 agaaatcctc gtgaacgaag aggatattaa gggctacaga gtgaccagca ccggcaagcc 8461 cgagttcacc aacctgaagg tgtaccacga catcaaggac attaccgccc ggaaagagat 8521 tattgagaac gccgagctgc tggatcagat tgccaagatc ctgaccatct accagagcag 8581 cgaggacatc caggaagaac tgaccaatct gaactccgag ctgacccagg aagagatcga 8641 gcagatctct aatctgaagg gctataccgg cacccacaac ctgagcctga aggccatcaa 8701 cctgatcctg gacgagctgt ggcacaccaa cgacaaccag atcgctatct tcaaccggct 8761 gaagctggtg cccaagaagg tggacctgtc ccagcagaaa gagatcccca ccaccctggt 8821 ggacgacttc atcctgagcc ccgtcgtgaa gagaagcttc atccagagca tcaaagtgat 8881 caacgccatc atcaagaagt acggcctgcc caacgacatc attatcgagc tggcccgcga 8941 gaagaactcc aaggacgccc agaaaatgat caacgagatg cagaagcgga accggcagac 9001 caacgagcgg atcgaggaaa tcatccggac caccggcaaa gagaacgcca agtacctgat 9061 cgagaagatc aagctgcacg acatgcagga aggcaagtgc ctgtacagcc tggaagccat 9121 ccctctggaa gatctgctga acaacccctt caactatgag gtggaccaca tcatccccag 9181 aagcgtgtcc ttcgacaaca gcttcaacaa caaggtgctc gtgaagcagg aagaagccag 9241 caagaagggc aaccggaccc cattccagta cctgagcagc agcgacagca agatcagcta 9301 cgaaaccttc aagaagcaca tcctgaatct ggccaagggc aagggcagaa tcagcaagac 9361 caagaaagag tatctgctgg aagaacggga catcaacagg ttctccgtgc agaaagactt 9421 catcaaccgg aacctggtgg ataccagata cgccaccaga ggcctgatga acctgctgcg 9481 gagctacttc agagtgaaca acctggacgt gaaagtgaag tccatcaatg gcggcttcac 9541 cagctttctg cggcggaagt ggaagtttaa gaaagagcgg aacaaggggt acaagcacca 9601 cgccgaggac gccctgatca ttgccaacgc cgatttcatc ttcaaagagt ggaagaaact 9661 ggacaaggcc aaaaaagtga tggaaaacca gatgttcgag gaaaagcagg ccgagagcat 9721 gcccgagatc gaaaccgagc aggagtacaa agagatcttc atcacccccc accagatcaa 9781 gcacattaag gacttcaagg actacaagta cagccaccgg gtggacaaga agcctaatag 9841 agagctgatt aacgacaccc tgtactccac ccggaaggac gacaagggca acaccctgat 9901 cgtgaacaat ctgaacggcc tgtacgacaa ggacaatgac aagctgaaaa agctgatcaa 9961 caagagcccc gaaaagctgc tgatgtacca ccacgacccc cagacctacc agaaactgaa 10021 gctgattatg gaacagtacg gcgacgagaa gaatcccctg tacaagtact acgaggaaac 10081 cgggaactac ctgaccaagt actccaaaaa ggacaacggc cccgtgatca agaagattaa 10141 gtattacggc aacaaactga acgcccatct ggacatcacc gacgactacc ccaacagcag 10201 aaacaaggtc gtgaagctgt ccctgaagcc ctacagattc gacgtgtacc tggacaatgg 10261 cgtgtacaag ttcgtgaccg tgaagaatct ggatgtgatc aaaaaagaaa actactacga 10321 agtgaatagc aagtgctatg aggaagctaa gaagctgaag aagatcagca accaggccga 10381 gtttatcgcc tccttctaca acaacgatct gatcaagatc aacggcgagc tgtatagagt 10441 gatcggcgtg aacaacgacc tgctgaaccg gatcgaagtg aacatgatcg acatcaccta 10501 ccgcgagtac ctggaaaaca tgaacgacaa gaggcccccc aggatcatta agacaatcgc 10561 ctccaagacc cagagcatta agaagtacag cacagacatt ctgggcaacc tgtatgaagt 10621 gaaatctaag aagcaccctc agatcatcaa aaagggcaaa aggccggcgg ccacgaaaaa 10681 ggccggccag gcaaaaaaga aaaagtaact cgagctagtt aaaagttttg ttactttata 10741 gaagaaattt tgagtttttg ttttttttta ataaataaat aaacataaat aaattgtttg 10801 ttgaatttat tattagtatg taagtgtaaa tataataaaa cttaatatct attcaaatta 10861 ataaataaac ctcgatatac agaccgataa aacacatgcg tcaattttac gcatgattat 10921 ctttaacgta cgtcacaata tgattatctt tctagggtta atctagtata cgcgtatgcg 10981 gccgcttaat taatccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta 11041 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat 11101 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt 11161 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta 11221 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc 11281 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa 11341 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa 11401 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct 11461 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac 11521 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc 11581 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc 11641 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg 11701 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga 11761 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag 11821 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta 11881 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag 11941 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg 12001 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac 12061 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc 12121 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag 12181 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc 12241 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac 12301 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc 12361 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg 12421 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag 12481 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc 12541 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac 12601 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag 12661 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac 12721 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg 12781 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc 12841 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact 12901 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg 12961 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa 13021 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt 13081 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg 13141 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa 13201 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt 13261 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata 13321 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac 13381 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa 13441 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc 13501 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg 13561 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca 13621 cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac 13681 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga 13741 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa 13801 acgacggcca gtgagcgcgc gtaatacgac tcactatagg gcgaattggg gcgcgccatt 13861 ctagattaac cctagaaaga tagtctgcgt aaaattgacg catgcattct tgaaatattg 13921 ctctctcttt ctaaatagcg cgaatccgtc gctgtgcatt taggacatct cagtcgccgc 13981 ttggagctcc cgtgaggcgt gcttgtcaat gcggtaagtg tcactgattt tgaactataa 14041 cgaccgcgtg agtcaaaatg acgcatgatt atcttttacg tgacttttaa gatttaactc 14101 atacgataat tatattgtta tttcatgttc tacttacgtg ataacttatt atatatatat 14161 tttcttgtta tagatatcaa ctagaatgct agcatgggcc cagatct //