LOCUS Exported 13713 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Modified version of lentiSAM v2, a lenti sgRNA cloning backbone with MS2 loops at tetraloop/stemloop 2, dCas9-VP64, and puro resistance marker. Contains BsmBI sites for insertion of spacer sequences.. ACCESSION . VERSION . KEYWORDS lentiSAM v2 (Puro) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 13713) TITLE Lenti CRISPR Activate JOURNAL Unpublished REFERENCE 2 (bases 1 to 13713) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..13713 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(47..66) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 238..617 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 619..817 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 772..792 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 835..1015 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 1062..1187 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1680..1913 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 2098..2142 /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" misc_feature 2440..2557 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2607..2847 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 2607..2627 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 2778..2797 /label=LKO.1 5' /note="Human U6 promoter, forward primer" misc_RNA 2893..2911 /label=MS2 stem loop /note="stem loop that binds the bacteriophage MS2 coat protein" misc_RNA 2963..2981 /label=MS2 stem loop /note="stem loop that binds the bacteriophage MS2 coat protein" promoter 3068..3279 /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" CDS 3296..3343 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" CDS 3344..7444 /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 N863A mutation in the HNH catalytic domain" /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKARGKSDNVPS EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD ATLIHQSITGLYETRIDLSQLGGD" CDS 7499..7519 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 7544..7693 /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 7715..7771 /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 7772..8368 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind 8265..8285 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 8405..8993 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(8458..8478) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(8876..8887) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(8996..9012) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(8997..9013) /label=pBluescriptKS /note="For pBluescript vector" LTR 9518..9698 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(9724..9741) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 9730..9954 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 10000..10428 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(10087..10106) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 10297..10318 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(10437..10457) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 10442..10771 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 10622..10757 /label=SV40 ori /note="SV40 origin of replication" primer_bind 10684..10703 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 10819..10866 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 10885..11259 /codon_start=1 /gene="Sh ble from Streptoalloteichus hindustanus" /product="antibiotic-binding protein" /label=BleoR /note="confers resistance to bleomycin, phleomycin, and Zeocin(TM)" /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR EFALRDPAGNCVHFVAEEQD" polyA_signal 11389..11510 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(11426..11445) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 11480..11499 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(11559..11575) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(11559..11575) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(11572..11594) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 11583..11599 /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)." promoter complement(11607..11637) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 11652..11673 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(11790..11807) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(11961..12549) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(12041..12060) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(12720..13580) /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 13343..13362 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(13581..13685) /gene="bla" /label=AmpR promoter ORIGIN 1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg 61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt 121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc 181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac 241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat 301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg 361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt 421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag 481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc 541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag 601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt 661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc 721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg 781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct 841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc 1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc 1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa 1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg 1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata 1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc 1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga 1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc 1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca 1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg 1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga 1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata 1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg 1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg 1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag 1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt 1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt 1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt 2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag 2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata 2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta 2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta 2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa 2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt 2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat 2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa 2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag 2581 agatccagtt tggttaatta gctagcgagg gcctatttcc catgattcct tcatatttgc 2641 atatacgata caaggctgtt agagagataa ttggaattaa tttgactgta aacacaaaga 2701 tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa 2761 aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct 2821 tggctttata tatcttgtgg aaaggacgaa acaccggaga cgggataccg tctctgtttt 2881 agagctaggc caacatgagg atcacccatg tctgcagggc ctagcaagtt aaaataaggc 2941 tagtccgtta tcaacttggc caacatgagg atcacccatg tctgcagggc caagtggcac 3001 cgagtcggtg ctttttttgg atcctaggtc ttgaaaggag tgggaattgg ctccggtgcc 3061 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc 3121 aattgatccg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac 3181 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg 3241 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg cgtacggcca ccatgaaaag 3301 gccggcggcc acgaaaaagg ccggccaggc aaaaaagaaa aaggacaaga agtacagcat 3361 cggcctggcc atcggcacca actctgtggg ctgggccgtg atcaccgacg agtacaaggt 3421 gcccagcaag aaattcaagg tgctgggcaa caccgaccgg cacagcatca agaagaacct 3481 gatcggagcc ctgctgttcg acagcggcga aacagccgag gccacccggc tgaagagaac 3541 cgccagaaga agatacacca gacggaagaa ccggatctgc tatctgcaag agatcttcag 3601 caacgagatg gccaaggtgg acgacagctt cttccacaga ctggaagagt ccttcctggt 3661 ggaagaggat aagaagcacg agcggcaccc catcttcggc aacatcgtgg acgaggtggc 3721 ctaccacgag aagtacccca ccatctacca cctgagaaag aaactggtgg acagcaccga 3781 caaggccgac ctgcggctga tctatctggc cctggcccac atgatcaagt tccggggcca 3841 cttcctgatc gagggcgacc tgaaccccga caacagcgac gtggacaagc tgttcatcca 3901 gctggtgcag acctacaacc agctgttcga ggaaaacccc atcaacgcca gcggcgtgga 3961 cgccaaggcc atcctgtctg ccagactgag caagagcaga cggctggaaa atctgatcgc 4021 ccagctgccc ggcgagaaga agaatggcct gttcggcaac ctgattgccc tgagcctggg 4081 cctgaccccc aacttcaaga gcaacttcga cctggccgag gatgccaaac tgcagctgag 4141 caaggacacc tacgacgacg acctggacaa cctgctggcc cagatcggcg accagtacgc 4201 cgacctgttt ctggccgcca agaacctgtc cgacgccatc ctgctgagcg acatcctgag 4261 agtgaacacc gagatcacca aggcccccct gagcgcctct atgatcaaga gatacgacga 4321 gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta 4381 caaagagatt ttcttcgacc agagcaagaa cggctacgcc ggctacattg acggcggagc 4441 cagccaggaa gagttctaca agttcatcaa gcccatcctg gaaaagatgg acggcaccga 4501 ggaactgctc gtgaagctga acagagagga cctgctgcgg aagcagcgga ccttcgacaa 4561 cggcagcatc ccccaccaga tccacctggg agagctgcac gccattctgc ggcggcagga 4621 agatttttac ccattcctga aggacaaccg ggaaaagatc gagaagatcc tgaccttccg 4681 catcccctac tacgtgggcc ctctggccag gggaaacagc agattcgcct ggatgaccag 4741 aaagagcgag gaaaccatca ccccctggaa cttcgaggaa gtggtggaca agggcgcttc 4801 cgcccagagc ttcatcgagc ggatgaccaa cttcgataag aacctgccca acgagaaggt 4861 gctgcccaag cacagcctgc tgtacgagta cttcaccgtg tataacgagc tgaccaaagt 4921 gaaatacgtg accgagggaa tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc 4981 catcgtggac ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga 5041 ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt 5101 caacgcctcc ctgggcacat accacgatct gctgaaaatt atcaaggaca aggacttcct 5161 ggacaatgag gaaaacgagg acattctgga agatatcgtg ctgaccctga cactgtttga 5221 ggacagagag atgatcgagg aacggctgaa aacctatgcc cacctgttcg acgacaaagt 5281 gatgaagcag ctgaagcggc ggagatacac cggctggggc aggctgagcc ggaagctgat 5341 caacggcatc cgggacaagc agtccggcaa gacaatcctg gatttcctga agtccgacgg 5401 cttcgccaac agaaacttca tgcagctgat ccacgacgac agcctgacct ttaaagagga 5461 catccagaaa gcccaggtgt ccggccaggg cgatagcctg cacgagcaca ttgccaatct 5521 ggccggcagc cccgccatta agaagggcat cctgcagaca gtgaaggtgg tggacgagct 5581 cgtgaaagtg atgggccggc acaagcccga gaacatcgtg atcgaaatgg ccagagagaa 5641 ccagaccacc cagaagggac agaagaacag ccgcgagaga atgaagcgga tcgaagaggg 5701 catcaaagag ctgggcagcc agatcctgaa agaacacccc gtggaaaaca cccagctgca 5761 gaacgagaag ctgtacctgt actacctgca gaatgggcgg gatatgtacg tggaccagga 5821 actggacatc aaccggctgt ccgactacga tgtggaccac atcgtgcctc agagctttct 5881 gaaggacgac tccatcgaca acaaggtgct gaccagaagc gacaaggccc ggggcaagag 5941 cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag aactactggc ggcagctgct 6001 gaacgccaag ctgattaccc agagaaagtt cgacaatctg accaaggccg agagaggcgg 6061 cctgagcgaa ctggataagg ccggcttcat caagagacag ctggtggaaa cccggcagat 6121 cacaaagcac gtggcacaga tcctggactc ccggatgaac actaagtacg acgagaatga 6181 caagctgatc cgggaagtga aagtgatcac cctgaagtcc aagctggtgt ccgatttccg 6241 gaaggatttc cagttttaca aagtgcgcga gatcaacaac taccaccacg cccacgacgc 6301 ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc tggaaagcga 6361 gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag atgatcgcca agagcgagca 6421 ggaaatcggc aaggctaccg ccaagtactt cttctacagc aacatcatga actttttcaa 6481 gaccgagatt accctggcca acggcgagat ccggaagcgg cctctgatcg agacaaacgg 6541 cgaaaccggg gagatcgtgt gggataaggg ccgggatttt gccaccgtgc ggaaagtgct 6601 gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa 6661 agagtctatc ctgcccaaga ggaacagcga taagctgatc gccagaaaga aggactggga 6721 ccctaagaag tacggcggct tcgacagccc caccgtggcc tattctgtgc tggtggtggc 6781 caaagtggaa aagggcaagt ccaagaaact gaagagtgtg aaagagctgc tggggatcac 6841 catcatggaa agaagcagct tcgagaagaa tcccatcgac tttctggaag ccaagggcta 6901 caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag tactccctgt tcgagctgga 6961 aaacggccgg aagagaatgc tggcctctgc cggcgaactg cagaagggaa acgaactggc 7021 cctgccctcc aaatatgtga acttcctgta cctggccagc cactatgaga agctgaaggg 7081 ctcccccgag gataatgagc agaaacagct gtttgtggaa cagcacaagc actacctgga 7141 cgagatcatc gagcagatca gcgagttctc caagagagtg atcctggccg acgctaatct 7201 ggacaaagtg ctgtccgcct acaacaagca ccgggataag cccatcagag agcaggccga 7261 gaatatcatc cacctgttta ccctgaccaa tctgggagcc cctgccgcct tcaagtactt 7321 tgacaccacc atcgaccgga agaggtacac cagcaccaaa gaggtgctgg acgccaccct 7381 gatccaccag agcatcaccg gcctgtacga gacacggatc gacctgtctc agctgggagg 7441 cgacagcgct ggaggaggtg gaagcggagg aggaggaagc ggaggaggag gtagcggacc 7501 taagaaaaag aggaaggtgg cggccgctgg atccggacgg gctgacgcat tggacgattt 7561 tgatctggat atgctgggaa gtgacgccct cgatgatttt gaccttgaca tgcttggttc 7621 ggatgccctt gatgactttg acctcgacat gctcggcagt gacgcccttg atgatttcga 7681 cctggacatg ctgattaact gtacaggcag tggagcaaca aacttctctc tgctgaaaca 7741 agccggagat gtcgaagaga atcctggacc gaccgagtac aagcccacgg tgcgcctcgc 7801 cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc 7861 cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga 7921 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc 7981 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat 8041 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg 8101 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggagtctc 8161 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc 8221 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta 8281 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg 8341 gtgcatgacc cgcaagcccg gtgcctgaac gcgttaagtc gaattcgata tcaagcttat 8401 cggtaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt 8461 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc 8521 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga 8581 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc 8641 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct 8701 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg 8761 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct 8821 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc 8881 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg 8941 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata 9001 ccgtcgacct cgagacctag aaaaacatgg agcaatcaca agtagcaata cagcagctac 9061 caatgctgat tgtgcctggc tagaagcaca agaggaggag gaggtgggtt ttccagtcac 9121 acctcaggta cctttaagac caatgactta caaggcagct gtagatctta gccacttttt 9181 aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag atatccttga 9241 tctgtggatc taccacacac aaggctactt ccctgattgg cagaactaca caccagggcc 9301 agggatcaga tatccactga cctttggatg gtgctacaag ctagtaccag ttgagcaaga 9361 gaaggtagaa gaagccaatg aaggagagaa cacccgcttg ttacaccctg tgagcctgca 9421 tgggatggat gacccggaga gagaagtatt agagtggagg tttgacagcc gcctagcatt 9481 tcatcacatg gcccgagagc tgcatccgga ctgtactggg tctctctggt tagaccagat 9541 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt 9601 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc 9661 cctcagaccc ttttagtcag tgtggaaaat ctctagcagg gcccgtttaa acccgctgat 9721 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt 9781 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat 9841 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg 9901 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg 9961 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat 10021 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag 10081 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc 10141 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc 10201 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt 10261 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa 10321 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg 10381 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa 10441 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag 10501 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag 10561 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc 10621 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt 10681 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg 10741 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt 10801 cggatctgat cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa 10861 tacgacaagg tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc 10921 gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac 10981 ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg 11041 gtccaggacc aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac 11101 gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg 11161 gccatgaccg agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc 11221 ggcaactgcg tgcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat 11281 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg 11341 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt 11401 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt 11461 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt 11521 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga 11581 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc 11641 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc 11701 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc 11761 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt 11821 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca 11881 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa 11941 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat 12001 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc 12061 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc 12121 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt 12181 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac 12241 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg 12301 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca 12361 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc 12421 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa 12481 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa 12541 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac 12601 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta 12661 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt 12721 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata 12781 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc 12841 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac 12901 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag 12961 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac 13021 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc 13081 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg 13141 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc 13201 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct 13261 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc 13321 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc 13381 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc 13441 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc 13501 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca 13561 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt 13621 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt 13681 ccgcgcacat ttccccgaaa agtgccacct gac //