LOCUS Exported 11589 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Lentiviral vector expressing Cas9 and an sgRNA targeting C1orf27. ACCESSION . VERSION . KEYWORDS LentiCRISPR-sgC1orf27-1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 11589) AUTHORS Wang T, Yu H, Hughes NW, Liu B, Kendirli A, Klein K, Chen WW, Lander ES, Sabatini DM TITLE Gene Essentiality Profiling Reveals Gene Networks and Synthetic Lethal Interactions with Oncogenic Ras. JOURNAL Cell. 2017 Feb 1. pii: S0092-8674(17)30061-2. doi: 10.1016/j.cell.2017.01.013. PUBMED 28162770 REFERENCE 2 (bases 1 to 11589) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..11589 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 5..231 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 232..412 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 459..584 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1077..1310 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1495..1539 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" promoter 1717..1957 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 1717..1737 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 1888..1907 /label=LKO.1 5' /note="Human U6 promoter, forward primer" misc_RNA 1986..2061 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" misc_feature 2109..2226 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2295..2506 /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" regulatory 2526..2535 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2535..2558 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 2565..2585 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 2610..6710 /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 6711..6758 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" CDS 6774..6830 /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 6831..7430 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(6831..6850) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 7327..7347 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 7446..8034 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(7499..7519) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(7917..7928) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 8106..8339 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 8417..8538 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(8454..8473) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 8508..8527 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 8578..8713 /label=SV40 ori /note="SV40 origin of replication" primer_bind 8640..8659 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(8733..8752) /label=T7 /note="T7 promoter, forward primer" promoter complement(8734..8752) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(8762..8779) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(8762..8778) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(8771..8793) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 8920..9375 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(9007..9026) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 9217..9238 /label=F1ori-F /note="F1 origin, forward primer" promoter 9401..9505 /gene="bla" /label=AmpR promoter CDS 9506..10366 /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(9724..9743) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 10537..11125 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 11026..11045 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 11279..11296 /label=L4440 /note="L4440 vector, forward primer" protein_bind 11413..11434 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 11449..11479 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 11487..11503 /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 11492..11514 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 11511..11527 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 11511..11527 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 11546..11566 /label=T3 /note="T3 promoter, forward primer" promoter 11548..11566 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" ORIGIN 1 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa 61 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac 121 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa 181 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc 241 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag 301 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct 361 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc 421 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc 481 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt 541 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga 601 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat 661 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt 721 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag 781 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa 841 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa 901 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg 961 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta 1021 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga 1081 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg 1141 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg 1201 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc 1261 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg 1321 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat 1381 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac 1441 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat 1501 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca 1561 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga 1621 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg 1681 tttcagaccc acctcccaac cccgagggga cccagagagg gcctatttcc catgattcct 1741 tcatatttgc atatacgata caaggctgtt agagagataa ttagaattaa tttgactgta 1801 aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt 1861 gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat 1921 ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccgagat tgtggaattt 1981 cacaggtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa 2041 aaagtggcac cgagtcggtg cttttttgaa ttctagatct tgagacaaat ggcagtattc 2101 atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta 2161 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa 2221 aattttcggg tttattacag ggacagcaga gatccacttt ggcgccggct cgagtggctc 2281 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg 2341 ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt 2401 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt 2461 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggtgt cgtgacgcgg 2521 gatccgccac catggattac aaagacgatg acgataagat ggccccaaag aagaagcgga 2581 aggtcggtat ccacggagtc ccagcagccg acaagaagta cagcatcggc ctggacatcg 2641 gcaccaactc tgtgggctgg gccgtgatca ccgacgagta caaggtgccc agcaagaaat 2701 tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa gaacctgatc ggagccctgc 2761 tgttcgacag cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc agaagaagat 2821 acaccagacg gaagaaccgg atctgctatc tgcaagagat cttcagcaac gagatggcca 2881 aggtggacga cagcttcttc cacagactgg aagagtcctt cctggtggaa gaggataaga 2941 agcacgagcg gcaccccatc ttcggcaaca tcgtggacga ggtggcctac cacgagaagt 3001 accccaccat ctaccacctg agaaagaaac tggtggacag caccgacaag gccgacctgc 3061 ggctgatcta tctggccctg gcccacatga tcaagttccg gggccacttc ctgatcgagg 3121 gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt catccagctg gtgcagacct 3181 acaaccagct gttcgaggaa aaccccatca acgccagcgg cgtggacgcc aaggccatcc 3241 tgtctgccag actgagcaag agcagacggc tggaaaatct gatcgcccag ctgcccggcg 3301 agaagaagaa tggcctgttc ggaaacctga ttgccctgag cctgggcctg acccccaact 3361 tcaagagcaa cttcgacctg gccgaggatg ccaaactgca gctgagcaag gacacctacg 3421 acgacgacct ggacaacctg ctggcccaga tcggcgacca gtacgccgac ctgtttctgg 3481 ccgccaagaa cctgtccgac gccatcctgc tgagcgacat cctgagagtg aacaccgaga 3541 tcaccaaggc ccccctgagc gcctctatga tcaagagata cgacgagcac caccaggacc 3601 tgaccctgct gaaagctctc gtgcggcagc agctgcctga gaagtacaaa gagattttct 3661 tcgaccagag caagaacggc tacgccggct acattgacgg cggagccagc caggaagagt 3721 tctacaagtt catcaagccc atcctggaaa agatggacgg caccgaggaa ctgctcgtga 3781 agctgaacag agaggacctg ctgcggaagc agcggacctt cgacaacggc agcatccccc 3841 accagatcca cctgggagag ctgcacgcca ttctgcggcg gcaggaagat ttttacccat 3901 tcctgaagga caaccgggaa aagatcgaga agatcctgac cttccgcatc ccctactacg 3961 tgggccctct ggccagggga aacagcagat tcgcctggat gaccagaaag agcgaggaaa 4021 ccatcacccc ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc cagagcttca 4081 tcgagcggat gaccaacttc gataagaacc tgcccaacga gaaggtgctg cccaagcaca 4141 gcctgctgta cgagtacttc accgtgtata acgagctgac caaagtgaaa tacgtgaccg 4201 agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc 4261 tgttcaagac caaccggaaa gtgaccgtga agcagctgaa agaggactac ttcaagaaaa 4321 tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga tcggttcaac gcctccctgg 4381 gcacatacca cgatctgctg aaaattatca aggacaagga cttcctggac aatgaggaaa 4441 acgaggacat tctggaagat atcgtgctga ccctgacact gtttgaggac agagagatga 4501 tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga caaagtgatg aagcagctga 4561 agcggcggag atacaccggc tggggcaggc tgagccggaa gctgatcaac ggcatccggg 4621 acaagcagtc cggcaagaca atcctggatt tcctgaagtc cgacggcttc gccaacagaa 4681 acttcatgca gctgatccac gacgacagcc tgacctttaa agaggacatc cagaaagccc 4741 aggtgtccgg ccagggcgat agcctgcacg agcacattgc caatctggcc ggcagccccg 4801 ccattaagaa gggcatcctg cagacagtga aggtggtgga cgagctcgtg aaagtgatgg 4861 gccggcacaa gcccgagaac atcgtgatcg aaatggccag agagaaccag accacccaga 4921 agggacagaa gaacagccgc gagagaatga agcggatcga agagggcatc aaagagctgg 4981 gcagccagat cctgaaagaa caccccgtgg aaaacaccca gctgcagaac gagaagctgt 5041 acctgtacta cctgcagaat gggcgggata tgtacgtgga ccaggaactg gacatcaacc 5101 ggctgtccga ctacgatgtg gaccatatcg tgcctcagag ctttctgaag gacgactcca 5161 tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg caagagcgac aacgtgccct 5221 ccgaagaggt cgtgaagaag atgaagaact actggcggca gctgctgaac gccaagctga 5281 ttacccagag aaagttcgac aatctgacca aggccgagag aggcggcctg agcgaactgg 5341 ataaggccgg cttcatcaag agacagctgg tggaaacccg gcagatcaca aagcacgtgg 5401 cacagatcct ggactcccgg atgaacacta agtacgacga gaatgacaag ctgatccggg 5461 aagtgaaagt gatcaccctg aagtccaagc tggtgtccga tttccggaag gatttccagt 5521 tttacaaagt gcgcgagatc aacaactacc accacgccca cgacgcctac ctgaacgccg 5581 tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga aagcgagttc gtgtacggcg 5641 actacaaggt gtacgacgtg cggaagatga tcgccaagag cgagcaggaa atcggcaagg 5701 ctaccgccaa gtacttcttc tacagcaaca tcatgaactt tttcaagacc gagattaccc 5761 tggccaacgg cgagatccgg aagcggcctc tgatcgagac aaacggcgaa accggggaga 5821 tcgtgtggga taagggccgg gattttgcca ccgtgcggaa agtgctgagc atgccccaag 5881 tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt cagcaaagag tctatcctgc 5941 ccaagaggaa cagcgataag ctgatcgcca gaaagaagga ctgggaccct aagaagtacg 6001 gcggcttcga cagccccacc gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg 6061 gcaagtccaa gaaactgaag agtgtgaaag agctgctggg gatcaccatc atggaaagaa 6121 gcagcttcga gaagaatccc atcgactttc tggaagccaa gggctacaaa gaagtgaaaa 6181 aggacctgat catcaagctg cctaagtact ccctgttcga gctggaaaac ggccggaaga 6241 gaatgctggc ctctgccggc gaactgcaga agggaaacga actggccctg ccctccaaat 6301 atgtgaactt cctgtacctg gccagccact atgagaagct gaagggctcc cccgaggata 6361 atgagcagaa acagctgttt gtggaacagc acaagcacta cctggacgag atcatcgagc 6421 agatcagcga gttctccaag agagtgatcc tggccgacgc taatctggac aaagtgctgt 6481 ccgcctacaa caagcaccgg gataagccca tcagagagca ggccgagaat atcatccacc 6541 tgtttaccct gaccaatctg ggagcccctg ccgccttcaa gtactttgac accaccatcg 6601 accggaagag gtacaccagc accaaagagg tgctggacgc caccctgatc caccagagca 6661 tcaccggcct gtacgagaca cggatcgacc tgtctcagct gggaggcgac aagcgtcctg 6721 ctgctactaa gaaagctggt caagctaaga aaaagaaagc tagcggcagc ggcgccacca 6781 acttcagcct gctgaagcag gccggcgacg tggaggagaa ccccggcccc atgaccgagt 6841 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg 6901 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg 6961 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg 7021 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag 7081 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg 7141 ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt 7201 tcctggccac cgtcggagtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg 7261 tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg 7321 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg 7381 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga acgcgttaag 7441 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg 7501 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt 7561 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg 7621 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc 7681 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc 7741 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc 7801 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc 7861 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg 7921 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc 7981 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcgtcgac 8041 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg 8101 gggactggaa gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg 8161 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact 8221 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg 8281 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag 8341 tacgtatagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat 8401 atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag 8461 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa 8521 actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc 8581 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt 8641 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc 8701 ttttttggag gcctagggac gtacccaatt cgccctatag tgagtcgtat tacgcgcgct 8761 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc 8821 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc 8881 gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat 8941 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag 9001 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc 9061 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc 9121 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt 9181 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa 9241 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg 9301 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat 9361 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt 9421 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct 9481 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc 9541 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa 9601 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg 9661 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt 9721 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg 9781 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac 9841 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc 9901 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa 9961 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc 10021 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt 10081 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga 10141 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa 10201 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa 10261 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa 10321 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt 10381 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt 10441 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg 10501 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt 10561 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca 10621 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac 10681 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac 10741 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct 10801 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg 10861 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca 10921 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt 10981 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta 11041 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc 11101 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc 11161 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa 11221 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag 11281 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg 11341 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga 11401 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat 11461 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag 11521 ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac aaaagctgga 11581 gctgcaagc //