LOCUS Exported 10238 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Express CHO codon-optimized dCas9 fused to VPR for CRISPR activation. 2A linked ZsGreen1 marker. ACCESSION . VERSION . KEYWORDS VPR_dCas9 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10238) AUTHORS Karottki KJC, Hefzi H, Xiong K, Shamie I, Hansen AH, Li S, Pedersen LE, Li S, Lee JS, Lee GM, Kildegaard HF, Lewis NE TITLE Awakening dormant glycosyltransferases in CHO cells with CRISPRa. JOURNAL Biotechnol Bioeng. 2020 Feb;117(2):593-598. doi: 10.1002/bit.27199. Epub 2019 Nov 12. PUBMED 31631317 REFERENCE 2 (bases 1 to 10238) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10238 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(287..1147) /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 910..929 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1148..1239) /gene="bla" /label=AmpR promoter primer_bind 1281..1300 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 1306..1325 /label=pENTR-R /note="pENTR vectors, reverse primer" enhancer 1538..1917 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 1918..2121 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2072..2092 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 2176..2185 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2209..2358 /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 2383..2403 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 2833..3189 /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 EDFSSIADMDFSALL" CDS 3208..3777 /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 3787..7887 /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 ATLIHQSITGLYETRIDLSQLGGD" CDS 7900..7920 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 7939..7992 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="EGRGSLLTCGDVEENPGP" CDS 7993..8685 /codon_start=1 /product="Zoanthus green fluorescent protein" /label=ZsGreen1 /note="mammalian codon-optimized" /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH AIASGSALP" polyA_signal 8830..9054 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(9529..9546) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(9529..9545) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(9644..9661) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(9815..165) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(9895..9914) /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc 61 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt 121 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc 181 ttttctacgg ggtctgacgc tcagtggaac gacgcgcgcg taactcacgt taagggattt 241 tggtcatgag cttgcgccgt cccgtcaagt cagcgtaatg ctctgcttac caatgcttaa 301 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc 361 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagc gctgcgatga 421 taccgcgaga accacgctca ccggctccgg atttatcagc aataaaccag ccagccggaa 481 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt 541 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccatcg 601 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc 661 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg 721 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag 781 cgctacataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt 841 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt 901 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac 961 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac 1021 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag 1081 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa 1141 tactcatatt cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga 1201 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggtcagt gttacaacca 1261 attaaccaat tctgaacatt atcgcgagcc catttatacc tgaatatggc tcataacacc 1321 ccttgtttgc ctggcggcag tagcgcggtg gtcccacctg accccatgcc gaactcagaa 1381 gtgaaacgcc gtagcgccga tggtagtgtg gggactcccc atgcgagagt agggaactgc 1441 caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgcc cgggctaatt 1501 atggggtgtc gcccttattc gactcagtcg gtgtgttgac attgattatt gactagttat 1561 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca 1621 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca 1681 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg 1741 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg 1801 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc 1861 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg 1921 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca 1981 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt 2041 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg 2101 gaggtctata taagcagagc tctctggcta actagagaac ccactgctta ctggcttatc 2161 gaaataagca gcgtcgccac catggaggcc agcggttccg gacgggctga cgcattggac 2221 gattttgatc tggatatgct gggaagtgac gccctcgatg attttgacct tgacatgctt 2281 ggttcggatg cccttgatga ctttgacctc gacatgctcg gcagtgacgc ccttgatgat 2341 ttcgacctgg acatgctgat taactctaga agttccggat ctccgaaaaa gaaacgcaaa 2401 gttggtagcc agtacctgcc cgacaccgac gaccggcacc ggatcgagga aaagcggaag 2461 cggacctacg agacattcaa gagcatcatg aagaagtccc ccttcagcgg ccccaccgac 2521 cctaggcctc cacctagaag aatcgccgtg cccagcagat ccagcgccag cgtgccaaaa 2581 cctgcccccc agccttaccc cttcaccagc agcctgagca ccatcaacta cgacgagttc 2641 cctaccatgg tgttccccag cggccagatc tctcaggcct ctgctctggc cccagcccct 2701 cctcaggtgc tgcctcaggc tcctgctcct gcaccagctc cagccatggt gtctgcactg 2761 gctcaggcac cagcacccgt gcctgtgctg gctcctggac ctccacaggc tgtggctcca 2821 ccagccccta aacctacaca ggccggcgag ggcacactgt ctgaagctct gctgcagctg 2881 cagttcgacg acgaggatct gggagccctg ctgggaaaca gcaccgatcc tgccgtgttc 2941 accgacctgg ccagcgtgga caacagcgag ttccagcagc tgctgaacca gggcatccct 3001 gtggcccctc acaccaccga gcccatgctg atggaatacc ccgaggccat cacccggctc 3061 gtgacaggcg ctcagaggcc tcctgatcca gctcctgccc ctctgggagc accaggcctg 3121 cctaatggac tgctgtctgg cgacgaggac ttcagctcta tcgccgatat ggatttctca 3181 gccttgctgg gctctggcag cggcagccgg gattccaggg aagggatgtt tttgccgaag 3241 cctgaggccg gctccgctat tagtgacgtg tttgagggcc gcgaggtgtg ccagccaaaa 3301 cgaatccggc catttcatcc tccaggaagt ccatgggcca accgcccact ccccgccagc 3361 ctcgcaccaa caccaaccgg tccagtacat gagccagtcg ggtcactgac cccggcacca 3421 gtccctcagc cactggatcc agcgcccgca gtgactcccg aggccagtca cctgttggag 3481 gatcccgatg aagagacgag ccaggctgtc aaagcccttc gggagatggc cgatactgtg 3541 attccccaga aggaagaggc tgcaatctgt ggccaaatgg acctttccca tccgccccca 3601 aggggccatc tggatgagct gacaaccaca cttgagtcca tgaccgagga tctgaacctg 3661 gactcacccc tgaccccgga attgaacgag attctggata ccttcctgaa cgacgagtgc 3721 ctcttgcatg ccatgcatat cagcacagga ctgtccatct tcgacacatc tctgtttaaa 3781 gcctatgaca agaaatactc catcggcctt gccatcggga ccaacagcgt gggttgggcc 3841 gtgatcactg acgaatacaa agtgccatcg aaaaagttta aggtgctggg caatactgac 3901 agacatagca tcaagaaaaa cctgattgga gccctcctgt tcgactccgg ggaaaccgcc 3961 gaagccactc gcctgaaacg gaccgcacgc cggaggtaca ctcgccggaa gaatagaatc 4021 tgctaccttc aagagatctt tagcaacgag atggccaaag tggacgattc gttcttccac 4081 cggctggagg agtcattcct ggtcgaagag gacaaaaagc atgagagaca tccgatcttc 4141 ggtaacattg tggatgaggt cgcctaccac gagaagtacc ccactatcta ccatttgcgc 4201 aagaagttgg tcgactcgac tgataaggcc gacctcagac tcatctacct cgctttggca 4261 cacatgatca agtttagggg tcacttcctg attgaagggg atctgaaccc agacaactcg 4321 gatgtggata aactgtttat ccagctggtc cagacttaca atcagctctt tgaggagaac 4381 ccgatcaatg cctccggcgt ggatgctaag gcaatcctgt ccgcacgctt gtcaaagagc 4441 cggcggctcg aaaatctgat cgcacagctt ccgggagaaa agaaaaacgg actgtttgga 4501 aaccttatcg ccctctcgct gggactcact cctaacttca aatccaattt tgatttggcc 4561 gaggatgcca agctgcagct gtcgaaggac acctacgacg atgacctcga caatcttctg 4621 gcccagatcg gggaccagta cgccgatctg ttcctcgcgg ccaaaaacct gtcagacgca 4681 atcttgctga gcgatatctt gagggtcaat accgaaatca ctaaggctcc attgtcagca 4741 tcgatgatca agaggtacga tgaacaccat caggacctga ccctgctcaa ggcactggtg 4801 cggcagcaac tgccggaaaa gtacaaggag atcttcttcg atcaaagcaa gaatgggtac 4861 gcagggtaca tcgatggtgg agcatcacaa gaagagttct acaaattcat caagcccatc 4921 cttgaaaaga tggacgggac ggaagaactg ctggtgaagc tgaatcggga agatctgctg 4981 cggaagcagc gcaccttcga caatggttcc atcccacacc aaatccatct cggcgaactg 5041 cacgcgatcc ttcgccggca ggaagatttc tacccgttct tgaaggataa cagagaaaag 5101 atcgagaaaa tcctgacctt tagaatcccg tactacgtgg gcccgttggc tcgcggaaac 5161 tcaagattcg cctggatgac tagaaaatcc gaagaaacga ttaccccgtg gaactttgaa 5221 gaggtcgtcg acaagggagc ctcggcacag tcgttcatcg aacggatgac caatttcgac 5281 aagaacctcc ccaacgaaaa ggtgctcccg aaacactcgc tgctgtatga gtacttcacg 5341 gtctacaacg agctgaccaa ggtgaagtac gtgaccgaag gaatgcggaa gcccgctttt 5401 ctgtccggtg aacagaaaaa ggccatcgtg gacctcctct tcaagactaa cagaaaggtg 5461 accgtgaaac agcttaaaga ggactacttc aagaagatcg agtgttttga ctccgtggaa 5521 atctcgggag tggaggatag gtttaacgct tcgctgggga cctaccatga ccttctcaag 5581 atcatcaagg ataaggattt cctggacaac gaagaaaacg aggacatcct tgaggacatc 5641 gtgttgacct tgaccctgtt tgaggatcgg gaaatgatcg aggagagact gaaaacttac 5701 gcgcatcttt tcgacgacaa agtgatgaag cagctgaaaa gaagaagata cactggctgg 5761 ggacggctgt cgaggaaact gattaacgga attagggata aacaaagcgg aaagactatc 5821 ctcgattttc tcaagtcgga cggcttcgcc aatcggaact ttatgcagct catccacgac 5881 gatagcctga ccttcaagga agatatccaa aaagcccaag tgtccggcca gggagatagc 5941 ttgcacgagc acattgctaa cctggccggt tcaccagcca ttaagaaggg aatcctccaa 6001 accgtgaagg tggtcgacga actcgtcaag gtgatgggcc gccataagcc agagaacatc 6061 gtgatcgaga tggctcgcga aaatcagact actcagaagg gtcaaaagaa ctcccgcgag 6121 cgcatgaagc gcatcgaaga aggaattaag gagctgggct cacaaatcct caaagaacat 6181 cctgtcgaga acacccaact tcagaacgag aaactgtacc tctactatct ccaaaatggc 6241 cgggacatgt acgtcgatca ggaattggat atcaaccgcc tctcagacta cgacgtggat 6301 gctatcgtgc cgcagtcgtt tctcaaagat gatagcatcg acaacaaggt cctgacgagg 6361 tcggataaga accgcgggaa atccgacaat gtgccttcgg aagaggtggt gaaaaagatg 6421 aagaactact ggaggcaatt gcttaatgcc aaactgatca cccagcgcaa attcgacaac 6481 cttactaagg ccgagcgcgg aggactgtcc gaactcgata aggcggggtt catcaaaaga 6541 caattggtcg aaactcggca aattaccaag catgtcgctc agatcctcga ctcgcgcatg 6601 aacactaagt atgatgagaa cgacaagctc attcgcgaag tgaaagtgat taccctcaaa 6661 tcaaagctgg tgtcggactt cagaaaagac tttcaattct acaaggtgcg ggagattaac 6721 aactaccacc acgcgcacga cgcctacctc aatgcagtgg tgggaaccgc cctcatcaag 6781 aagtatccaa agctcgaaag cgagttcgtg tacggggact ataaagtgta cgatgtgcgg 6841 aaaatgatcg caaagagcga gcaagagatc ggcaaagcaa cggccaaata cttcttctac 6901 agcaacatta tgaacttttt caagaccgag attaccctgg cgaacggaga aatccggaag 6961 cggcctttga tcgaaacgaa tggggaaact ggagaaatcg tgtgggacaa gggcagagac 7021 tttgcgaccg tgagaaaggt gctgtccatg ccacaagtca acatcgtgaa gaaaactgaa 7081 gtgcagactg gaggattttc caaggagtca atccttccga agcgcaacag cgacaagctg 7141 atcgccagga agaaggattg ggaccccaag aagtacggtg gttttgattc acctactgtc 7201 gcttacagcg tgctggtggt ggccaaggtg gaaaagggga agtcaaagaa attgaagtcg 7261 gtgaaggaat tgctggggat tactatcatg gagaggagca gcttcgaaaa gaatcccatc 7321 gacttcttgg aggccaaggg atacaaagag gtgaagaaag acctgatcat taagctgccg 7381 aagtactcct tgttcgaact ggaaaacggc agaaagcgga tgctggcctc cgccggagag 7441 ctgcagaagg gtaacgagct ggctctgccc agcaaatacg tgaatttcct ctacctggcc 7501 tcccattacg agaagctcaa gggaagcccg gaggataatg agcaaaaaca actgttcgtc 7561 gagcagcaca agcactacct cgacgagatc atcgaacaaa tctccgagtt ctcgaagcgg 7621 gtgattctgg ccgacgcaaa ccttgataaa gtcctcagcg cctacaacaa gcaccgcgac 7681 aaaccgatca gagaacaagc tgagaacatc atccacctgt tcacgctgac taacttggga 7741 gcgcctgccg ccttcaagta cttcgacacc actatcgatc ggaaacggta cacttccact 7801 aaggaagtgc tggacgcaac cctgatccat cagtccatca ccggacttta tgagactcgg 7861 atcgatctca gccagcttgg cggtgattca agagctgacc cgaagaagaa gcgcaaggtc 7921 agacgtcatg gcagtggaga gggcagagga agtctgctaa catgcggtga cgtcgaggag 7981 aatcctggcc caatggccca gtccaagcac ggcctgacca aggagatgac catgaagtac 8041 cgcatggagg gctgcgtgga cggccacaag ttcgtgatca ccggcgaggg catcggctac 8101 cccttcaagg gcaagcaggc catcaacctg tgcgtggtgg agggcggccc cttgcccttc 8161 gccgaggaca tcttgtccgc cgccttcatg tacggcaacc gcgtgttcac cgagtacccc 8221 caggacatcg tcgactactt caagaactcc tgccccgccg gctacacctg ggaccgctcc 8281 ttcctgttcg aggacggcgc cgtgtgcatc tgcaacgccg acatcaccgt gagcgtggag 8341 gagaactgca tgtaccacga gtccaagttc tacggcgtga acttccccgc cgacggcccc 8401 gtgatgaaga agatgaccga caactgggag ccctcctgcg agaagatcat ccccgtgccc 8461 aagcagggca tcttgaaggg cgacgtgagc atgtacctgc tgctgaagga cggtggccgc 8521 ttgcgctgcc agttcgacac cgtgtacaag gccaagtccg tgccccgcaa gatgcccgac 8581 tggcacttca tccagcacaa gctgacccgc gaggaccgca gcgacgccaa gaaccagaag 8641 tggcacctga ccgagcacgc catcgcctcc ggctccgcct tgcccaagct tccgcggagc 8701 catggcttcc cgccggcggt ggcggcgcag gatgatggca cgctgcccat gtcttgtgcc 8761 caggagagcg ggatggaccg tcaccctgca gcctgtgctt ctgctaggat caatgtgtag 8821 acacagtctc tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt 8881 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat 8941 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg 9001 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggacttgc 9061 gtagtgagtc gaataagggc gacacaaaat ttattctaaa tgcataataa atactgataa 9121 catcttatag tttgtattat attttgtatt atcgttgaca tgtataattt tgatatcaaa 9181 aactgatttt ccctttatta ttttcgagat ttattttctt aattctcttt aacaaactag 9241 aaatattgta tatacaaaaa atcataaata atagatgaat agtttaatta taggtgttca 9301 tcaatcgaaa aagcaacgta tcttatttaa agtgcgttgc ttttttctca tttataaggt 9361 taaataattc tcatatatca agcaaagtga caggcgccct taaatattct gacaaatgct 9421 ctttccctaa actcccccca taaaaaaacc cgccgaagcg ggtttttacg ttatttgcgg 9481 attaacgatt actcgttatc agaaccgccc agggggcccg agcttaagac tggccgtcgt 9541 tttacaacac agaaagagtt tgtagaaacg caaaaaggcc atccgtcagg ggccttctgc 9601 ttagtttgat gcctggcagt tccctactct cgccttccgc ttcctcgctc actgactcgc 9661 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 9721 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 9781 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 9841 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 9901 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 9961 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 10021 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 10081 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 10141 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 10201 taggcggtgc tacagagttc ttgaagtggt gggctaac //