LOCUS Exported 9756 bp ds-DNA circular SYN 17-OCT-2022 DEFINITION Plasmid ensures constitutive expression of the C-terminal fragment of split ERbeta (aa 447-501), fused to the catalytically inactive Cas9 (dCas9) protein, a flexible GS linker and the SV40 NLS signal at the N-terminus. ACCESSION . VERSION . KEYWORDS dCas9-NLS-gs10-cERb447 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9756) AUTHORS Rihtar E, Lebar T, Lainscek D, Kores K, Lesnik S, Bren U, Jerala R TITLE Chemically inducible split protein regulators for mammalian cells JOURNAL Nat Chem Biol. 2022 REFERENCE 2 (bases 1 to 9756) AUTHORS . TITLE Direct Submission JOURNAL Exported Oct 17, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9756 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(55..74) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 246..625 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 626..829 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 780..800 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 874..893 /label=T7 /note="T7 promoter, forward primer" promoter 874..892 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 964..981 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 982..5079 /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 ATLIHQSITGLYETRIDLSQLGG" CDS 5086..5106 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" promoter complement(5320..5338) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(5321..5338) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(5358..5375) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 5364..5588 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 5634..6062 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5721..5740) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5931..5952 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(6071..6091) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 6076..6405 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 6256..6391 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6318..6337 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 6472..7266 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(6526..6545) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 7136..7155 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 7440..7561 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(7477..7496) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 7531..7550 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(7610..7626) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7610..7626) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(7623..7645) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 7634..7650 /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(7658..7688) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7703..7724 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(7841..7858) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(8012..8600) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(8092..8111) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8771..9631) /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 9394..9413 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(9632..9736) /gene="bla" /label=AmpR promoter ORIGIN 1 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat 61 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc 121 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat 181 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac 241 gcgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca 301 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc 361 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat 421 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt 481 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc 541 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta 601 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg 661 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt 721 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac 781 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac 841 tagagaaccc actgcttact ggcttatcga aattaatacg actcactata gggagaccca 901 agcttggtac cgagctcgga tccactagta acggccgcca gtgtgctgga attcgccacc 961 atgcaccacc accaccacca cgacaagaag tacagcatcg gcctggccat cggcaccaac 1021 tctgtgggct gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg 1081 ctgggcaaca ccgaccggca cagcatcaag aagaacctga tcggcgccct gctgttcgac 1141 agcggagaaa cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga 1201 cggaagaacc ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac 1261 gacagcttct tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag 1321 cggcacccca tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc 1381 atctaccacc tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc 1441 tatctggccc tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg 1501 aaccccgaca acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag 1561 ctgttcgagg aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc 1621 agactgagca agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag 1681 aatggcctgt tcggcaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc 1741 aacttcgacc tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac 1801 ctggacaacc tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag 1861 aacctgtccg acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag 1921 gcccccctga gcgcctctat gatcaagaga tacgacgagc accaccagga cctgaccctg 1981 ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag 2041 agcaagaacg gctacgccgg ctacatcgat ggcggagcca gccaggaaga gttctacaag 2101 ttcatcaagc ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac 2161 agagaggacc tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc ccaccagatc 2221 cacctgggag agctgcacgc cattctgcgg cggcaggaag atttttaccc attcctgaag 2281 gacaaccggg aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct 2341 ctggccaggg gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc 2401 ccctggaact tcgaggaagt ggtggacaag ggcgccagcg cccagagctt catcgagcgg 2461 atgaccaact tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg 2521 tacgagtact tcaccgtgta caacgagctg accaaagtga aatacgtgac cgagggaatg 2581 agaaagcccg ccttcctgag cggcgagcag aaaaaagcca tcgtggacct gctgttcaag 2641 accaaccgga aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc 2701 ttcgactccg tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac 2761 cacgatctgc tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac 2821 attctggaag atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa 2881 cggctgaaaa cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggcgg 2941 agatacaccg gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag 3001 tccggcaaga caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg 3061 cagctgatcc acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc 3121 ggccagggcg atagcctgca cgagcacatt gccaatctgg ccggcagccc cgccattaag 3181 aagggcatcc tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat gggccggcac 3241 aagcccgaga acatcgtgat cgaaatggcc agagagaacc agaccaccca gaagggacag 3301 aagaacagcc gcgagagaat gaagcggatc gaagagggca tcaaagagct gggcagccag 3361 atcctgaaag aacaccccgt ggaaaacacc cagctgcaga acgagaagct gtacctgtac 3421 tacctgcaga atgggcggga tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc 3481 gactacgatg tggacgctat cgtgcctcag agctttctga aggacgactc catcgataac 3541 aaagtgctga ctcggagcga caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag 3601 gtcgtgaaga agatgaagaa ctactggcgc cagctgctga atgccaagct gattacccag 3661 aggaagttcg acaatctgac caaggccgag agaggcggcc tgagcgaact ggataaggcc 3721 ggcttcatca agagacagct ggtggaaacc cggcagatca caaagcacgt ggcacagatc 3781 ctggactccc ggatgaacac taagtacgac gagaacgaca aactgatccg ggaagtgaaa 3841 gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga aggatttcca gttttacaaa 3901 gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga 3961 accgccctga tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag 4021 gtgtacgacg tgcggaagat gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc 4081 aagtacttct tctacagcaa catcatgaac tttttcaaga ccgagattac cctggccaac 4141 ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg aaacaggcga gatcgtgtgg 4201 gataagggcc gggactttgc caccgtgcgg aaagtgctgt ctatgcccca agtgaatatc 4261 gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag agtctatcct gcccaagagg 4321 aacagcgaca agctgatcgc cagaaagaag gactgggacc ctaagaagta cggcggcttc 4381 gacagcccca ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc 4441 aagaaactga agagtgtgaa agagctgctg gggatcacca tcatggaaag aagcagcttc 4501 gagaagaatc ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg 4561 atcatcaagc tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg 4621 gcctctgccg gcgaactgca gaagggaaac gaactggccc tgccctccaa atatgtgaac 4681 ttcctgtacc tggccagcca ctatgagaag ctgaagggct cccccgagga taatgagcag 4741 aaacagctgt ttgtggaaca gcacaaacac tacctggacg agatcatcga gcagatcagc 4801 gagttctcca agagagtgat cctggccgac gctaatctgg acaaggtgct gagcgcctac 4861 aacaagcaca gagacaagcc tatcagagag caggccgaga atatcatcca cctgtttacc 4921 ctgaccaatc tgggagcccc tgccgccttc aagtactttg acaccaccat cgaccggaag 4981 aggtacacca gcaccaaaga ggtgctggac gccaccctga tccaccagag catcaccggc 5041 ctgtacgaga cacggatcga cctgtctcag ctgggaggct ctgatccaaa aaagaagaga 5101 aaggtaggcg gctctggcgg cggctccgga ggctcttcca gccagcaaca atctatgcga 5161 ttggcgaact tgcttatgct gctcagtcac gtaaggcatg ccagcaataa agggatggaa 5221 catcttctta atatgaagtg taagaatgtg gtgcccgttt acgatctcct gctggaaatg 5281 ctcaacgcac acgttctcag gtaatctaga gggccctatt ctatagtgtc acctaaatgc 5341 tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc 5401 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa 5461 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg 5521 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg 5581 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc 5641 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact 5701 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc 5761 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt 5821 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc 5881 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt 5941 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat 6001 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa 6061 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc 6121 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc 6181 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg 6241 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat 6301 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc 6361 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct 6421 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa 6481 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac 6541 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg 6601 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag 6661 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt 6721 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg 6781 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg 6841 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga 6901 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag 6961 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat 7021 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt 7081 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg 7141 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt 7201 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc 7261 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac 7321 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg 7381 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca 7441 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 7501 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 7561 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt 7621 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa 7681 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac 7741 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg 7801 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc 7861 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 7921 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 7981 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 8041 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 8101 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 8161 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 8221 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 8281 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 8341 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 8401 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat 8461 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 8521 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc 8581 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 8641 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 8701 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 8761 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 8821 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 8881 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat 8941 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 9001 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 9061 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 9121 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 9181 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 9241 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 9301 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 9361 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 9421 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 9481 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 9541 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 9601 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 9661 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 9721 aaataggggt tccgcgcaca tttccccgaa aagtgc //