LOCUS Exported 13570 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Lentiviral vector expressing Cas9-GFP. ACCESSION . VERSION . KEYWORDS pLenti-Cas9-GFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 13570) 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 13570) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..13570 /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 3847..3922 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" misc_feature 3970..4087 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 4156..4367 /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" regulatory 4387..4396 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 4396..4419 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 4426..4446 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 4471..8571 /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 8572..8619 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" CDS 8635..8691 /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 8692..9408 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(8737..8758) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(8998..9017) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 9345..9366 /label=EGFP-C /note="EGFP, forward primer" misc_feature 9427..10015 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(9480..9500) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(9898..9909) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 10087..10320 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 10398..10519 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(10435..10454) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 10489..10508 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 10559..10694 /label=SV40 ori /note="SV40 origin of replication" primer_bind 10621..10640 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(10714..10733) /label=T7 /note="T7 promoter, forward primer" promoter complement(10715..10733) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(10743..10760) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(10743..10759) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(10752..10774) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 10901..11356 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(10988..11007) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 11198..11219 /label=F1ori-F /note="F1 origin, forward primer" promoter 11382..11486 /gene="bla" /label=AmpR promoter CDS 11487..12347 /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(11705..11724) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 12518..13106 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 13007..13026 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 13260..13277 /label=L4440 /note="L4440 vector, forward primer" protein_bind 13394..13415 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 13430..13460 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 13468..13484 /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 13473..13495 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 13492..13508 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 13492..13508 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 13527..13547 /label=T3 /note="T3 promoter, forward primer" promoter 13529..13547 /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 acaccggaga cggttgtaaa 1981 tgagcacaca aaatacacat gctaaaatat tatattctat gacctttata aaatcaacca 2041 aaatcttctt tttaataact ttagtatcaa taattagaat ttttatgttc ctttttgcaa 2101 acttttaata aaaatgagca aaataaaaaa acgctagttt tagtaactcg cgttgttttc 2161 ttcaccttta ataatagcta ctccaccact tgttcctaag cggtcagctc ctgcttcaat 2221 cattttttga gcatcttcaa atgttctaac tccaccagct gctttaacta aagcattgtc 2281 tttaacaact gacttcatta gtttaacatc ttcaaatgtt gcacctgatt ttgaaaatcc 2341 tgttgatgtt ttaacaaatt ctaatccagc ttcaacagct atttcacaag ctttcatgat 2401 ttcttctttt gttaataaac aattttccat aatacattta acaacatgtg atccagctgc 2461 tttttttaca gctttcatgt cttctaaaac taattcataa tttttgtctt ttaatgcacc 2521 aatatttaat accatatcaa tttctgttgc accatcttta attgcttcag aaacttcgaa 2581 tgcttttgta gctgttgtgc atgcacctag aggaaaacct acaacatttg ttattcctac 2641 atttgtgcct tttaataatt ctttacaata gcttgttcaa tatgaattaa cacaaactgt 2701 tgcaaaatca aattcaattg cttcatcaca taattgttta atttcagctt tcgtagcatc 2761 ttgttttaat aatgtgtgat ctatatattt gtttagtttc attttttctc ctatatattc 2821 atttttaatt ttaattcttt aataatttcg tctactttaa ctttagcgtt ttgaacagat 2881 tcaccaacac ctataaaata aatttttagt ttaggttcag ttccacttgg gcgaacagca 2941 aatcatgact tatcttctaa ataaaatttt agtaagtctt gtcctggcat attatacatt 3001 ccatcgatgt agtcttcaac attaacaact ttaagtccag caatttgagt taagggtgtt 3061 gctctcaatg atttcattaa tggttcaatt tttaatttct tttcttctgg tttaaaattc 3121 aagtttaaag tgaaagtgta atatgcaccc atttctttaa ataaatcttc taaatagtct 3181 actaatgttt tattttgttt tttataaaat caagcagcct ctgctattaa tatagaagct 3241 tgtattccat ctttatctct agctgagtca tcaattacat atccataact ttcttcataa 3301 gcaaaaacaa aatttaatcc gttatcttct tctttagcaa tttctctacc cattcattta 3361 aatccagtta aagtttttac aatattaact ccatattttt catgagcgat tctatcaccc 3421 aaatcacttg ttacaaaact tgaatataga gccggatttt ttggaatgct atttaagcgt 3481 tttagatttg ataattttca atcaattaaa attggtcctg tttgatttcc atctaatctt 3541 acaaaatgac catcatgttt tattgccatt ccaaatctgt cagcatctgg gtcattcata 3601 ataataatat ctgcatcatg tttaatacca tattcaagcg gtatttttca tgcaggatca 3661 aattctggat ttggatttac aacattttta aatgtttcat cttcaaatgc atgctcttca 3721 acctcaataa cgttatatcc tgattcacgt aatatttttg gggtaaattt agttcctgtt 3781 ccattaactg cgctaaaaat aatttttaaa tcttttttag cttcttgctc ttttttgtac 3841 gtctctgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga 3901 aaaagtggca ccgagtcggt gcttttttga attctagatc ttgagacaaa tggcagtatt 3961 catccacaat tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt 4021 agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca 4081 aaattttcgg gtttattaca gggacagcag agatccactt tggcgccggc tcgagtggct 4141 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag 4201 gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg 4261 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag 4321 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggtg tcgtgacgcg 4381 ggatccgcca ccatggatta caaagacgat gacgataaga tggccccaaa gaagaagcgg 4441 aaggtcggta tccacggagt cccagcagcc gacaagaagt acagcatcgg cctggacatc 4501 ggcaccaact ctgtgggctg ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa 4561 ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga agaacctgat cggagccctg 4621 ctgttcgaca gcggcgaaac agccgaggcc acccggctga agagaaccgc cagaagaaga 4681 tacaccagac ggaagaaccg gatctgctat ctgcaagaga tcttcagcaa cgagatggcc 4741 aaggtggacg acagcttctt ccacagactg gaagagtcct tcctggtgga agaggataag 4801 aagcacgagc ggcaccccat cttcggcaac atcgtggacg aggtggccta ccacgagaag 4861 taccccacca tctaccacct gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg 4921 cggctgatct atctggccct ggcccacatg atcaagttcc ggggccactt cctgatcgag 4981 ggcgacctga accccgacaa cagcgacgtg gacaagctgt tcatccagct ggtgcagacc 5041 tacaaccagc tgttcgagga aaaccccatc aacgccagcg gcgtggacgc caaggccatc 5101 ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc tgatcgccca gctgcccggc 5161 gagaagaaga atggcctgtt cggaaacctg attgccctga gcctgggcct gacccccaac 5221 ttcaagagca acttcgacct ggccgaggat gccaaactgc agctgagcaa ggacacctac 5281 gacgacgacc tggacaacct gctggcccag atcggcgacc agtacgccga cctgtttctg 5341 gccgccaaga acctgtccga cgccatcctg ctgagcgaca tcctgagagt gaacaccgag 5401 atcaccaagg cccccctgag cgcctctatg atcaagagat acgacgagca ccaccaggac 5461 ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg agaagtacaa agagattttc 5521 ttcgaccaga gcaagaacgg ctacgccggc tacattgacg gcggagccag ccaggaagag 5581 ttctacaagt tcatcaagcc catcctggaa aagatggacg gcaccgagga actgctcgtg 5641 aagctgaaca gagaggacct gctgcggaag cagcggacct tcgacaacgg cagcatcccc 5701 caccagatcc acctgggaga gctgcacgcc attctgcggc ggcaggaaga tttttaccca 5761 ttcctgaagg acaaccggga aaagatcgag aagatcctga ccttccgcat cccctactac 5821 gtgggccctc tggccagggg aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa 5881 accatcaccc cctggaactt cgaggaagtg gtggacaagg gcgcttccgc ccagagcttc 5941 atcgagcgga tgaccaactt cgataagaac ctgcccaacg agaaggtgct gcccaagcac 6001 agcctgctgt acgagtactt caccgtgtat aacgagctga ccaaagtgaa atacgtgacc 6061 gagggaatga gaaagcccgc cttcctgagc ggcgagcaga aaaaggccat cgtggacctg 6121 ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga aagaggacta cttcaagaaa 6181 atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg 6241 ggcacatacc acgatctgct gaaaattatc aaggacaagg acttcctgga caatgaggaa 6301 aacgaggaca ttctggaaga tatcgtgctg accctgacac tgtttgagga cagagagatg 6361 atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg acaaagtgat gaagcagctg 6421 aagcggcgga gatacaccgg ctggggcagg ctgagccgga agctgatcaa cggcatccgg 6481 gacaagcagt ccggcaagac aatcctggat ttcctgaagt ccgacggctt cgccaacaga 6541 aacttcatgc agctgatcca cgacgacagc ctgaccttta aagaggacat ccagaaagcc 6601 caggtgtccg gccagggcga tagcctgcac gagcacattg ccaatctggc cggcagcccc 6661 gccattaaga agggcatcct gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg 6721 ggccggcaca agcccgagaa catcgtgatc gaaatggcca gagagaacca gaccacccag 6781 aagggacaga agaacagccg cgagagaatg aagcggatcg aagagggcat caaagagctg 6841 ggcagccaga tcctgaaaga acaccccgtg gaaaacaccc agctgcagaa cgagaagctg 6901 tacctgtact acctgcagaa tgggcgggat atgtacgtgg accaggaact ggacatcaac 6961 cggctgtccg actacgatgt ggaccatatc gtgcctcaga gctttctgaa ggacgactcc 7021 atcgacaaca aggtgctgac cagaagcgac aagaaccggg gcaagagcga caacgtgccc 7081 tccgaagagg tcgtgaagaa gatgaagaac tactggcggc agctgctgaa cgccaagctg 7141 attacccaga gaaagttcga caatctgacc aaggccgaga gaggcggcct gagcgaactg 7201 gataaggccg gcttcatcaa gagacagctg gtggaaaccc ggcagatcac aaagcacgtg 7261 gcacagatcc tggactcccg gatgaacact aagtacgacg agaatgacaa gctgatccgg 7321 gaagtgaaag tgatcaccct gaagtccaag ctggtgtccg atttccggaa ggatttccag 7381 ttttacaaag tgcgcgagat caacaactac caccacgccc acgacgccta cctgaacgcc 7441 gtcgtgggaa ccgccctgat caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc 7501 gactacaagg tgtacgacgt gcggaagatg atcgccaaga gcgagcagga aatcggcaag 7561 gctaccgcca agtacttctt ctacagcaac atcatgaact ttttcaagac cgagattacc 7621 ctggccaacg gcgagatccg gaagcggcct ctgatcgaga caaacggcga aaccggggag 7681 atcgtgtggg ataagggccg ggattttgcc accgtgcgga aagtgctgag catgccccaa 7741 gtgaatatcg tgaaaaagac cgaggtgcag acaggcggct tcagcaaaga gtctatcctg 7801 cccaagagga acagcgataa gctgatcgcc agaaagaagg actgggaccc taagaagtac 7861 ggcggcttcg acagccccac cgtggcctat tctgtgctgg tggtggccaa agtggaaaag 7921 ggcaagtcca agaaactgaa gagtgtgaaa gagctgctgg ggatcaccat catggaaaga 7981 agcagcttcg agaagaatcc catcgacttt ctggaagcca agggctacaa agaagtgaaa 8041 aaggacctga tcatcaagct gcctaagtac tccctgttcg agctggaaaa cggccggaag 8101 agaatgctgg cctctgccgg cgaactgcag aagggaaacg aactggccct gccctccaaa 8161 tatgtgaact tcctgtacct ggccagccac tatgagaagc tgaagggctc ccccgaggat 8221 aatgagcaga aacagctgtt tgtggaacag cacaagcact acctggacga gatcatcgag 8281 cagatcagcg agttctccaa gagagtgatc ctggccgacg ctaatctgga caaagtgctg 8341 tccgcctaca acaagcaccg ggataagccc atcagagagc aggccgagaa tatcatccac 8401 ctgtttaccc tgaccaatct gggagcccct gccgccttca agtactttga caccaccatc 8461 gaccggaaga ggtacaccag caccaaagag gtgctggacg ccaccctgat ccaccagagc 8521 atcaccggcc tgtacgagac acggatcgac ctgtctcagc tgggaggcga caagcgtcct 8581 gctgctacta agaaagctgg tcaagctaag aaaaagaaag ctagcggcag cggcgccacc 8641 aacttcagcc tgctgaagca ggccggcgac gtggaggaga accccggccc catggtgtct 8701 aagggcgaag agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta 8761 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg 8821 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc 8881 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac 8941 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac 9001 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc 9061 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag 9121 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag 9181 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac 9241 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc 9301 acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag 9361 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta aacgcgttaa 9421 gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat 9481 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct 9541 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag 9601 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc 9661 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc 9721 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct 9781 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg 9841 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg 9901 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg 9961 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcgtcga 10021 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg 10081 ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg 10141 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac 10201 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt 10261 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca 10321 gtacgtatag tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa 10381 tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata 10441 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca 10501 aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccat 10561 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt 10621 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg 10681 cttttttgga ggcctaggga cgtacccaat tcgccctata gtgagtcgta ttacgcgcgc 10741 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat 10801 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat 10861 cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca 10921 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta 10981 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt 11041 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac 11101 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt 11161 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga 11221 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg 11281 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata 11341 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt 11401 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc 11461 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc 11521 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa 11581 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg 11641 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag 11701 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc 11761 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta 11821 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg 11881 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca 11941 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac 12001 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat 12061 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg 12121 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata 12181 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta 12241 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa 12301 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag 12361 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg 12421 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact 12481 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg 12541 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc 12601 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata 12661 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta 12721 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc 12781 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg 12841 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac 12901 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg 12961 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt 13021 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct 13081 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg 13141 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata 13201 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca 13261 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc 13321 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg 13381 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta 13441 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca 13501 gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg 13561 agctgcaagc //