LOCUS Exported 11316 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION A guide RNA targeting TMPRSS2 in a lentiviral plasmid co-expressing Cas9 and TagBFP2. ACCESSION . VERSION . KEYWORDS TMPRSS2 g1 + Cas9 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 11316) TITLE CRISPR plasmids to knock out SARS-CoV-2 host genes JOURNAL Unpublished REFERENCE 2 (bases 1 to 11316) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..11316 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 132..151 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 385..402 /label=L4440 /note="L4440 vector, forward primer" protein_bind 519..540 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 555..585 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 593..609 /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)." primer_bind 598..620 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 617..633 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 617..633 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 652..672 /label=T3 /note="T3 promoter, forward primer" promoter 654..672 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 700..926 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 927..1107 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 1154..1279 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1772..2005 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." promoter complement(2191..2431) /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind complement(2241..2260) /label=LKO.1 5' /note="Human U6 promoter, forward primer" primer_bind complement(2411..2431) /label=hU6-F /note="Human U6 promoter, forward primer" misc_feature 2438..2555 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2624..2835 /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" regulatory 2855..2864 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2864..2887 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 2894..2914 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 2939..7039 /codon_start=1 /product="Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=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 7040..7087 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" CDS 7103..7159 /codon_start=1 /product="2A peptide from porcine teschovirus-1 polyprotein" /label=P2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="ATNFSLLKQAGDVEENPGP" CDS 7160..7861 /codon_start=1 /product="monomeric blue fluorescent protein" /label=TagBFP /note="mammalian codon-optimized" /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK LVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC DLPSKLGHKLN" misc_feature 7868..8456 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(7921..7941) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(8339..8350) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 8528..8761 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 8839..8960 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(8876..8895) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 8930..8949 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 9000..9135 /label=SV40 ori /note="SV40 origin of replication" primer_bind 9062..9081 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(9155..9174) /label=T7 /note="T7 promoter, forward primer" promoter complement(9156..9174) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(9184..9201) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(9184..9200) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(9193..9215) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 9342..9797 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(9429..9448) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 9639..9660 /label=F1ori-F /note="F1 origin, forward primer" promoter 9823..9927 /gene="bla" /label=AmpR promoter CDS 9928..10788 /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(10146..10165) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 10959..231 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" ORIGIN 1 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa 61 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg 121 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga 181 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc 241 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct 301 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct 361 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca 421 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg 481 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat 541 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc 601 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac 661 cctcactaaa gggaacaaaa gctggagctg caagcttaat gtagtcttat gcaatactct 721 tgtagtcttg caacatggta acgatgagtt agcaacatgc cttacaagga gagaaaaagc 781 accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac 841 agacgggtct gacatggatt ggacgaacca ctgaattgcc gcattgcaga gatattgtat 901 ttaagtgcct agctcgatac ataaacgggt ctctctggtt agaccagatc tgagcctggg 961 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc 1021 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct 1081 tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg 1141 gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg 1201 cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag 1261 agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat 1321 tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag 1381 ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca 1441 aatactggga cagctacaac catcccttca gacaggatca gaagaactta gatcattata 1501 taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga 1561 agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc 1621 cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata 1681 aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag 1741 tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag 1801 cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat 1861 tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc 1921 tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa 1981 gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcgccattt 2041 gtctcaagat ctagaattca aaaaaagcac cgactcggtg ccactttgct gtttccagca 2101 aagttgataa cggactagcc ttatttaaac ttgctatgct gtttccagca tagctcttaa 2161 accgcaaatg ccgtccaatg ccggtgtttc gtcctttcca caagatatat aaagccaaga 2221 aatcgaaata ctttcaagtt acggtaagca tatgatagtc cattttaaaa cataatttta 2281 aaactgcaaa ctacccaaga aattattact ttctacgtca cgtattttgt actaatatct 2341 ttgtgtttac agtcaaatta attctaatta tctctctaac agccttgtat cgtatatgca 2401 aatatgaagg aatcatggga aataggccct cccacaattt taaaagaaaa ggggggattg 2461 gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag 2521 aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg gacagcagag 2581 atccactttg gcgccggctc gagtggctcc ggtgcccgtc agtgggcaga gcgcacatcg 2641 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg 2701 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt 2761 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt 2821 gccgccagaa cacaggtgtc gtgacgcggg atccgccacc atggattaca aagacgatga 2881 cgataagatg gccccaaaga agaagcggaa ggtcggtatc cacggagtcc cagcagccga 2941 caagaagtac agcatcggcc tggacatcgg caccaactct gtgggctggg ccgtgatcac 3001 cgacgagtac aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg accggcacag 3061 catcaagaag aacctgatcg gagccctgct gttcgacagc ggcgaaacag ccgaggccac 3121 ccggctgaag agaaccgcca gaagaagata caccagacgg aagaaccgga tctgctatct 3181 gcaagagatc ttcagcaacg agatggccaa ggtggacgac agcttcttcc acagactgga 3241 agagtccttc ctggtggaag aggataagaa gcacgagcgg caccccatct tcggcaacat 3301 cgtggacgag gtggcctacc acgagaagta ccccaccatc taccacctga gaaagaaact 3361 ggtggacagc accgacaagg ccgacctgcg gctgatctat ctggccctgg cccacatgat 3421 caagttccgg ggccacttcc tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga 3481 caagctgttc atccagctgg tgcagaccta caaccagctg ttcgaggaaa accccatcaa 3541 cgccagcggc gtggacgcca aggccatcct gtctgccaga ctgagcaaga gcagacggct 3601 ggaaaatctg atcgcccagc tgcccggcga gaagaagaat ggcctgttcg gaaacctgat 3661 tgccctgagc ctgggcctga cccccaactt caagagcaac ttcgacctgg ccgaggatgc 3721 caaactgcag ctgagcaagg acacctacga cgacgacctg gacaacctgc tggcccagat 3781 cggcgaccag tacgccgacc tgtttctggc cgccaagaac ctgtccgacg ccatcctgct 3841 gagcgacatc ctgagagtga acaccgagat caccaaggcc cccctgagcg cctctatgat 3901 caagagatac gacgagcacc accaggacct gaccctgctg aaagctctcg tgcggcagca 3961 gctgcctgag aagtacaaag agattttctt cgaccagagc aagaacggct acgccggcta 4021 cattgacggc ggagccagcc aggaagagtt ctacaagttc atcaagccca tcctggaaaa 4081 gatggacggc accgaggaac tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca 4141 gcggaccttc gacaacggca gcatccccca ccagatccac ctgggagagc tgcacgccat 4201 tctgcggcgg caggaagatt tttacccatt cctgaaggac aaccgggaaa agatcgagaa 4261 gatcctgacc ttccgcatcc cctactacgt gggccctctg gccaggggaa acagcagatt 4321 cgcctggatg accagaaaga gcgaggaaac catcaccccc tggaacttcg aggaagtggt 4381 ggacaagggc gcttccgccc agagcttcat cgagcggatg accaacttcg ataagaacct 4441 gcccaacgag aaggtgctgc ccaagcacag cctgctgtac gagtacttca ccgtgtataa 4501 cgagctgacc aaagtgaaat acgtgaccga gggaatgaga aagcccgcct tcctgagcgg 4561 cgagcagaaa aaggccatcg tggacctgct gttcaagacc aaccggaaag tgaccgtgaa 4621 gcagctgaaa gaggactact tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg 4681 cgtggaagat cggttcaacg cctccctggg cacataccac gatctgctga aaattatcaa 4741 ggacaaggac ttcctggaca atgaggaaaa cgaggacatt ctggaagata tcgtgctgac 4801 cctgacactg tttgaggaca gagagatgat cgaggaacgg ctgaaaacct atgcccacct 4861 gttcgacgac aaagtgatga agcagctgaa gcggcggaga tacaccggct ggggcaggct 4921 gagccggaag ctgatcaacg gcatccggga caagcagtcc ggcaagacaa tcctggattt 4981 cctgaagtcc gacggcttcg ccaacagaaa cttcatgcag ctgatccacg acgacagcct 5041 gacctttaaa gaggacatcc agaaagccca ggtgtccggc cagggcgata gcctgcacga 5101 gcacattgcc aatctggccg gcagccccgc cattaagaag ggcatcctgc agacagtgaa 5161 ggtggtggac gagctcgtga aagtgatggg ccggcacaag cccgagaaca tcgtgatcga 5221 aatggccaga gagaaccaga ccacccagaa gggacagaag aacagccgcg agagaatgaa 5281 gcggatcgaa gagggcatca aagagctggg cagccagatc ctgaaagaac accccgtgga 5341 aaacacccag ctgcagaacg agaagctgta cctgtactac ctgcagaatg ggcgggatat 5401 gtacgtggac caggaactgg acatcaaccg gctgtccgac tacgatgtgg accatatcgt 5461 gcctcagagc tttctgaagg acgactccat cgacaacaag gtgctgacca gaagcgacaa 5521 gaaccggggc aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta 5581 ctggcggcag ctgctgaacg ccaagctgat tacccagaga aagttcgaca atctgaccaa 5641 ggccgagaga ggcggcctga gcgaactgga taaggccggc ttcatcaaga gacagctggt 5701 ggaaacccgg cagatcacaa agcacgtggc acagatcctg gactcccgga tgaacactaa 5761 gtacgacgag aatgacaagc tgatccggga agtgaaagtg atcaccctga agtccaagct 5821 ggtgtccgat ttccggaagg atttccagtt ttacaaagtg cgcgagatca acaactacca 5881 ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc 5941 taagctggaa agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat 6001 cgccaagagc gagcaggaaa tcggcaaggc taccgccaag tacttcttct acagcaacat 6061 catgaacttt ttcaagaccg agattaccct ggccaacggc gagatccgga agcggcctct 6121 gatcgagaca aacggcgaaa ccggggagat cgtgtgggat aagggccggg attttgccac 6181 cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac 6241 aggcggcttc agcaaagagt ctatcctgcc caagaggaac agcgataagc tgatcgccag 6301 aaagaaggac tgggacccta agaagtacgg cggcttcgac agccccaccg tggcctattc 6361 tgtgctggtg gtggccaaag tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga 6421 gctgctgggg atcaccatca tggaaagaag cagcttcgag aagaatccca tcgactttct 6481 ggaagccaag ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc 6541 cctgttcgag ctggaaaacg gccggaagag aatgctggcc tctgccggcg aactgcagaa 6601 gggaaacgaa ctggccctgc cctccaaata tgtgaacttc ctgtacctgg ccagccacta 6661 tgagaagctg aagggctccc ccgaggataa tgagcagaaa cagctgtttg tggaacagca 6721 caagcactac ctggacgaga tcatcgagca gatcagcgag ttctccaaga gagtgatcct 6781 ggccgacgct aatctggaca aagtgctgtc cgcctacaac aagcaccggg ataagcccat 6841 cagagagcag gccgagaata tcatccacct gtttaccctg accaatctgg gagcccctgc 6901 cgccttcaag tactttgaca ccaccatcga ccggaagagg tacaccagca ccaaagaggt 6961 gctggacgcc accctgatcc accagagcat caccggcctg tacgagacac ggatcgacct 7021 gtctcagctg ggaggcgaca agcgtcctgc tgctactaag aaagctggtc aagctaagaa 7081 aaagaaagct agcggcagcg gcgccaccaa cttcagcctg ctgaagcagg ccggcgacgt 7141 ggaggagaac cccggcccca tgagcgagct gattaaggag aacatgcaca tgaagctgta 7201 catggagggc accgtggaca accatcactt caagtgcaca tccgagggcg aaggcaagcc 7261 ctacgagggc acccagacca tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc 7321 cttcgacatc ctggctacta gcttcctcta cggcagcaag accttcatca accacaccca 7381 gggcatcccc gacttcttca agcagtcctt ccctgagggc ttcacatggg agagagtcac 7441 cacatacgaa gacgggggcg tgctgaccgc tacccaggac accagcctcc aggacggctg 7501 cctcatctac aacgtcaaga tcagaggggt gaacttcaca tccaacggcc ctgtgatgca 7561 gaagaaaaca ctcggctggg aggccttcac cgaaacgctg taccccgctg acggcggcct 7621 ggaaggcaga aacgacatgg ccctgaagct cgtgggcggg agccatctga tcgcaaacgc 7681 caagaccaca tatagatcca agaaacccgc taagaacctc aagatgcctg gcgtctacta 7741 tgtggactac agactggaaa gaatcaagga ggccaacaac gagacctacg tcgagcagca 7801 cgaggtggca gtggccagat actgcgacct ccctagcaaa ctggggcaca agcttaattg 7861 agtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta 7921 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc 7981 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga 8041 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac 8101 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc 8161 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc 8221 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg 8281 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc 8341 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc 8401 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcgtcg 8461 actttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag 8521 gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact 8581 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca 8641 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg 8701 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc 8761 agtacgtata gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga 8821 atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat 8881 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc 8941 aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca 9001 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt 9061 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag 9121 gcttttttgg aggcctaggg acgtacccaa ttcgccctat agtgagtcgt attacgcgcg 9181 ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa 9241 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga 9301 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc 9361 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct 9421 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg 9481 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga 9541 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt 9601 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg 9661 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc 9721 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat 9781 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt 9841 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg 9901 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt 9961 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta 10021 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc 10081 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa 10141 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc 10201 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt 10261 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact 10321 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac 10381 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata 10441 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta 10501 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg 10561 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat 10621 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt 10681 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga 10741 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa 10801 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag 10861 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac 10921 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc 10981 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat 11041 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat 11101 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct 11161 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt 11221 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg 11281 gggggttcgt gcacacagcc cagcttggag cgaacg //