LOCUS Exported 13895 bp ds-DNA circular SYN 24-SEP-2018 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pSECC SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 13895) AUTHORS FSR TITLE Direct Submission JOURNAL Exported Sep 24, 2018 from SnapGene 4.2.5 http://www.snapgene.com FEATURES Location/Qualifiers source 1..13895 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 237..616 /note="CMV enhancer" /note="human cytomegalovirus immediate early enhancer" promoter 618..816 /note="CMV promoter" /note="human cytomegalovirus (CMV) immediate early promoter" LTR 834..1014 /label=5' LTR (truncated) /note="5' LTR (truncated)" /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 1061..1186 /label=HIV-1 Psi /note="HIV-1 Psi" /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1685..1918 /label=RRE /note="RRE" /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." misc_feature 2445..2562 /label=cPPT/CTS /note="cPPT/CTS" /note="central polypurine tract and central termination sequence of HIV-1" misc_feature 2625..2654 /label=pAF Gibson /note="pAF Gibson" promoter 2655..2895 /note="U6 promoter" /note="RNA polymerase III promoter for human U6 snRNA" misc_feature 2900..4784 /label=Filler /note="Filler" misc_RNA 4785..4860 /label=gRNA scaffold /note="gRNA scaffold" /note="guide RNA scaffold for the CRISPR/Cas9 system" misc_feature 4908..5025 /label=cPPT/CTS /note="cPPT/CTS" /note="central polypurine tract and central termination sequence of HIV-1" misc_feature 5069..5098 /label=pBF Gibson /note="pBF Gibson" promoter 5124..5335 /note="EF-1-alpha core promoter" /note="core promoter for human elongation factor EF-1-alpha" misc_feature 5350..5379 /label=gBF Gibson /note="gBF Gibson" CDS 5383..5406 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG(R) epitope tag, followed by an enterokinas /note="FLAG" /translation="DYKDDDDK" CDS 5413..5433 /codon_start=1 /product="nuclear localization signal of SV40 large T antigen" /label=nuclear localization signal of SV40 large T ant /note="SV40 NLS" /translation="PKKKRKV" CDS 5458..9558 /codon_start=1 /product="Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=Cas9 (Csn1) endonuclease from the Streptococcus /note="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 9559..9606 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=bipartite nuclear localization signal from nucl /note="nucleoplasmin NLS" /translation="KRPAATKKAGQAKKKK" CDS 9622..9678 /codon_start=1 /product="2A peptide from porcine teschovirus-1 polyprotein" /label=2A peptide from porcine teschovirus-1 polyprotein /note="P2A" /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="ATNFSLLKQAGDVEENPGP" CDS 9682..9702 /codon_start=1 /product="nuclear localization signal of SV40 large T antigen" /label=nuclear localization signal of SV40 large T ant /note="SV40 NLS" /translation="PKKKRKV" CDS 9703..10731 /codon_start=1 /product="site-specific recombinase" /label=site-specific recombinase /note="Cre" /note="Cre is a site-specific recombinase from bacteriophage P1. Recombination occurs at loxP sequences." /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED GD" misc_feature 10732..10761 /label=gBR Gibson /note="gBR Gibson" misc_feature 10784..11372 /label=WPRE /note="WPRE" /note="woodchuck hepatitis virus posttranscriptional regulatory element" CDS complement(11255..11266) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa recognition and cleavage site /note="Factor Xa site" /translation="IEGR" primer_bind complement(11375..11391) /label=KS primer /note="KS primer" /note="common sequencing primer, one of multiple similar variants" LTR 11901..12081 /label=5' LTR (truncated) /note="5' LTR (truncated)" /note="truncated 5' long terminal repeat (LTR) from HIV-1" rep_origin complement(12143..12731) /direction=LEFT /label=ori /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(12902..13762) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=bla /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(13763..13867) /gene="bla" /label=bla promoter /note="AmpR promoter" ORIGIN 1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga 61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg 121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct 181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca 241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata 301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga 361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt 421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt 481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca 541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt 601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt 661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca 721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg 781 cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc 841 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 901 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 961 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg 1021 cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg 1081 gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat 1141 tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg 1201 gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa 1261 attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct 1321 gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac 1381 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca 1441 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa 1501 aagtaagacc accgcacagc aagcggccgg ccgcgctgat cttcagacct ggaggaggag 1561 atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat 1621 taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg 1681 gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt 1741 caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca 1801 atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca 1861 agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg 1921 ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt 1981 ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag 2041 aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag 2101 aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta 2161 acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag 2221 gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac 2281 cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag 2341 aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac ggatcggcac 2401 tgcgtgcgcc aattctgcag acaaatggca gtattcatcc acaattttaa aagaaaaggg 2461 gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa 2521 actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac 2581 agcagagatc cagtttggtt agtaccgggc ccgggtcgac tagtgatcag tgtgagggag 2641 tgtaaagctg gtttgagggc ctatttccca tgattccttc atatttgcat atacgataca 2701 aggctgttag agagataatt agaattaatt tgactgtaaa cacaaagata ttagtacaaa 2761 atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta 2821 aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttatata 2881 tcttgtggaa aggacgaaac accggagacg gttgtaaatg agcacacaaa atacacatgc 2941 taaaatatta tattctatga cctttataaa atcaaccaaa atcttctttt taataacttt 3001 agtatcaata attagaattt ttatgttcct ttttgcaaac ttttaataaa aatgagcaaa 3061 ataaaaaaac gctagtttta gtaactcgcg ttgttttctt cacctttaat aatagctact 3121 ccaccacttg ttcctaagcg gtcagctcct gcttcaatca ttttttgagc atcttcaaat 3181 gttctaactc caccagctgc tttaactaaa gcattgtctt taacaactga cttcattagt 3241 ttaacatctt caaatgttgc acctgatttt gaaaatcctg ttgatgtttt aacaaattct 3301 aatccagctt caacagctat ttcacaagct ttcatgattt cttcttttgt taataaacaa 3361 ttttccataa tacatttaac aacatgtgat ccagctgctt tttttacagc tttcatgtct 3421 tctaaaacta attcataatt tttgtctttt aatgcaccaa tatttaatac catatcaatt 3481 tctgttgcac catctttaat tgcttcagaa acttcgaatg cttttgtagc tgttgtgcat 3541 gcacctagag gaaaacctac aacatttgtt attcctacat ttgtgccttt taataattct 3601 ttacaatagc ttgttcaata tgaattaaca caaactgttg caaaatcaaa ttcaattgct 3661 tcatcacata attgtttaat ttcagctttc gtagcatctt gttttaataa tgtgtgatct 3721 atatatttgt ttagtttcat tttttctcct atatattcat ttttaatttt aattctttaa 3781 taatttcgtc tactttaact ttagcgtttt gaacagattc accaacacct ataaaataaa 3841 tttttagttt aggttcagtt ccacttgggc gaacagcaaa tcatgactta tcttctaaat 3901 aaaattttag taagtcttgt cctggcatat tatacattcc atcgatgtag tcttcaacat 3961 taacaacttt aagtccagca atttgagtta agggtgttgc tctcaatgat ttcattaatg 4021 gttcaatttt taatttcttt tcttctggtt taaaattcaa gtttaaagtg aaagtgtaat 4081 atgcacccat ttctttaaat aaatcttcta aatagtctac taatgtttta ttttgttttt 4141 tataaaatca agcagcctct gctattaata tagaagcttg tattccatct ttatctctag 4201 ctgagtcatc aattacatat ccataacttt cttcataagc aaaaacaaaa tttaatccgt 4261 tatcttcttc tttagcaatt tctctaccca ttcatttaaa tccagttaaa gtttttacaa 4321 tattaactcc atatttttca tgagcgattc tatcacccaa atcacttgtt acaaaacttg 4381 aatatagagc cggatttttt ggaatgctat ttaagcgttt tagatttgat aattttcaat 4441 caattaaaat tggtcctgtt tgatttccat ctaatcttac aaaatgacca tcatgtttta 4501 ttgccattcc aaatctgtca gcatctgggt cattcataat aataatatct gcatcatgtt 4561 taataccata ttcaagcggt atttttcatg caggatcaaa ttctggattt ggatttacaa 4621 catttttaaa tgtttcatct tcaaatgcat gctcttcaac ctcaataacg ttatatcctg 4681 attcacgtaa tatttttggg gtaaatttag ttcctgttcc attaactgcg ctaaaaataa 4741 tttttaaatc ttttttagct tcttgctctt ttttgtacgt ctctgtttta gagctagaaa 4801 tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc 4861 ttttttgaat tctagatctt gagacaaatg gcagtattca tccacaattt taaaagaaaa 4921 ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata 4981 caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg 5041 gacagcagag atccactttg gcgccggcaa accgctgttc ctaggaatcc cgaggccttc 5101 gagtggctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt 5161 ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg taaactggga 5221 aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag 5281 tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggtgtc 5341 gtgacgcggg acccgacatt agcgctacag cttaagcgga tggattacaa agacgatgac 5401 gataagatgg ccccaaagaa gaagcggaag gtcggtatcc acggagtccc agcagccgac 5461 aagaagtaca gcatcggcct ggacatcggc accaactctg tgggctgggc cgtgatcacc 5521 gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga ccggcacagc 5581 atcaagaaga acctgatcgg agccctgctg ttcgacagcg gcgaaacagc cgaggccacc 5641 cggctgaaga gaaccgccag aagaagatac accagacgga agaaccggat ctgctatctg 5701 caagagatct tcagcaacga gatggccaag gtggacgaca gcttcttcca cagactggaa 5761 gagtccttcc tggtggaaga ggataagaag cacgagcggc accccatctt cggcaacatc 5821 gtggacgagg tggcctacca cgagaagtac cccaccatct accacctgag aaagaaactg 5881 gtggacagca ccgacaaggc cgacctgcgg ctgatctatc tggccctggc ccacatgatc 5941 aagttccggg gccacttcct gatcgagggc gacctgaacc ccgacaacag cgacgtggac 6001 aagctgttca tccagctggt gcagacctac aaccagctgt tcgaggaaaa ccccatcaac 6061 gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg 6121 gaaaatctga tcgcccagct gcccggcgag aagaagaatg gcctgttcgg aaacctgatt 6181 gccctgagcc tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc 6241 aaactgcagc tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc 6301 ggcgaccagt acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg 6361 agcgacatcc tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc 6421 aagagatacg acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag 6481 ctgcctgaga agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac 6541 attgacggcg gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag 6601 atggacggca ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag 6661 cggaccttcg acaacggcag catcccccac cagatccacc tgggagagct gcacgccatt 6721 ctgcggcggc aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag 6781 atcctgacct tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc 6841 gcctggatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg 6901 gacaagggcg cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg 6961 cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac 7021 gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc 7081 gagcagaaaa aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag 7141 cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc 7201 gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag 7261 gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc 7321 ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg 7381 ttcgacgaca aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg 7441 agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc 7501 ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg 7561 acctttaaag aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag 7621 cacattgcca atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag 7681 gtggtggacg agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa 7741 atggccagag agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag 7801 cggatcgaag agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa 7861 aacacccagc tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg 7921 tacgtggacc aggaactgga catcaaccgg ctgtccgact acgatgtgga ccatatcgtg 7981 cctcagagct ttctgaagga cgactccatc gacaacaagg tgctgaccag aagcgacaag 8041 aaccggggca agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac 8101 tggcggcagc tgctgaacgc caagctgatt acccagagaa agttcgacaa tctgaccaag 8161 gccgagagag gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg 8221 gaaacccggc agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag 8281 tacgacgaga atgacaagct gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg 8341 gtgtccgatt tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac 8401 cacgcccacg acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct 8461 aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc 8521 gccaagagcg agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc 8581 atgaactttt tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg 8641 atcgagacaa acggcgaaac cggggagatc gtgtgggata agggccggga ttttgccacc 8701 gtgcggaaag tgctgagcat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca 8761 ggcggcttca gcaaagagtc tatcctgccc aagaggaaca gcgataagct gatcgccaga 8821 aagaaggact gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct 8881 gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag 8941 ctgctgggga tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg 9001 gaagccaagg gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc 9061 ctgttcgagc tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag 9121 ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat 9181 gagaagctga agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac 9241 aagcactacc tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg 9301 gccgacgcta atctggacaa agtgctgtcc gcctacaaca agcaccggga taagcccatc 9361 agagagcagg ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc 9421 gccttcaagt actttgacac caccatcgac cggaagaggt acaccagcac caaagaggtg 9481 ctggacgcca ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg 9541 tctcagctgg gaggcgacaa gcgtcctgct gctactaaga aagctggtca agctaagaaa 9601 aagaaagcta gcggcagcgg cgccaccaac ttcagcctgc tgaagcaggc cggcgacgtg 9661 gaggagaacc ccggccccat gcccaagaag aagaggaagg tgtccaattt actgaccgta 9721 caccaaaatt tgcctgcatt accggtcgat gcaacgagtg atgaggttcg caagaacctg 9781 atggacatgt tcagggatcg ccaggcgttt tctgagcata cctggaaaat gcttctgtcc 9841 gtttgccggt cgtgggcggc atggtgcaag ttgaataacc ggaaatggtt tcccgcagaa 9901 cctgaagatg ttcgcgatta tcttctatat cttcaggcgc gcggtctggc agtaaaaact 9961 atccagcaac atttgggcca gctaaacatg cttcatcgtc ggtccgggct gccacgacca 10021 agtgacagca atgctgtttc actggttatg cggcggatcc gaaaagaaaa cgttgatgcc 10081 ggtgaacgtg caaaacaggc tctagcgttc gaacgcactg atttcgacca ggttcgttca 10141 ctcatggaaa atagcgatcg ctgccaggat atacgtaatc tggcatttct ggggattgct 10201 tataacaccc tgttacgtat agccgaaatt gccaggatca gggttaaaga tatctcacgt 10261 actgacggtg ggagaatgtt aatccatatt ggcagaacga aaacgctggt tagcaccgca 10321 ggtgtagaga aggcacttag cctgggggta actaaactgg tcgagcgatg gatttccgta 10381 tctggtgtag ctgatgatcc gaataactac ctgttttgcc gggtcagaaa aaatggtgtt 10441 gccgcgccat ctgccaccag ccagctatca actcgcgccc tggaagggat ttttgaagca 10501 actcatcgat tgatttacgg cgctaaggat gactctggtc agagatacct ggcctggtct 10561 ggacacagtg cccgtgtcgg agccgcgcga gatatggccc gcgctggagt ttcaataccg 10621 gagatcatgc aagctggtgg ctggaccaat gtaaatattg tcatgaacta tatccgtaac 10681 ctggatagtg aaacaggggc aatggtgcgc ctgctggaag atggcgatta gaaacgttgt 10741 tgtttggggt tgaattactc ttctagaccg cggcgcgccc gataatcaac ctctggatta 10801 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg 10861 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc 10921 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca 10981 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac 11041 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact 11101 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc 11161 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg 11221 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc 11281 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac 11341 gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgtcgacctc gatcgagacc 11401 tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct 11461 ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa 11521 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac 11581 tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca 11641 cacaaggcta cttccctgat tggcagaact acacaccagg gccagggatc agatatccac 11701 tgacctttgg atggtgctac aagctagtac cagttgagca agagaaggta gaagaagcca 11761 atgaaggaga gaacacccgc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 11821 agagagaagt attagagtgg aggtttgaca gccgcctagc atttcatcac atggcccgag 11881 agctgcatcc ggactgtact gggtctctct ggttagacca gatctgagcc tgggagctct 11941 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag 12001 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt 12061 cagtgtggaa aatctctagc agcatgtgag caaaaggcca gcaaaaggcc aggaaccgta 12121 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 12181 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 12241 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 12301 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 12361 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 12421 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 12481 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 12541 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct 12601 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 12661 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa 12721 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa 12781 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt 12841 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca 12901 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca 12961 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc 13021 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa 13081 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc 13141 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca 13201 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat 13261 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag 13321 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac 13381 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt 13441 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt 13501 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc 13561 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat 13621 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca 13681 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga 13741 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg 13801 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg 13861 ttccgcgcac atttccccga aaagtgccac ctgac //