LOCUS Exported 14085 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION 3rd generation lenti vector encoding dCAS9-VP64 with 2A Blast resistance marker (EF1a-NLS-dCas9(N863)-VP64-2A-Blast-WPRE). ACCESSION . VERSION . KEYWORDS lenti dCAS-VP64_Blast SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 14085) AUTHORS Konermann S, Brigham MD, Trevino AE, Joung J, Abudayyeh OO, Barcena C, Hsu PD, Habib N, Gootenberg JS, Nishimasu H, Nureki O, Zhang F TITLE Genome-scale transcriptional activation by an engineered CRISPR-Cas9 complex. JOURNAL Nature. 2014 Dec 10. doi: 10.1038/nature14136. PUBMED 25494202 REFERENCE 2 (bases 1 to 14085) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..14085 /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 2687..3865 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 2918..3856 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 3813..3833 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" CDS 3881..3928 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" CDS 3929..8029 /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 8084..8104 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 8129..8278 /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 8300..8353 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="EGRGSLLTCGDVEENPGP" CDS 8354..8752 /codon_start=1 /gene="Aspergillus terreus BSD" /product="blasticidin S deaminase" /label=BSD /note="confers resistance to blasticidin" /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" misc_feature 8777..9365 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(8830..8850) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(9248..9259) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(9368..9384) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(9369..9385) /label=pBluescriptKS /note="For pBluescript vector" LTR 9890..10070 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(10096..10113) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 10102..10326 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 10372..10800 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(10459..10478) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 10669..10690 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(10809..10829) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 10814..11143 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 10994..11129 /label=SV40 ori /note="SV40 origin of replication" primer_bind 11056..11075 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 11191..11238 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 11257..11631 /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 11761..11882 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(11798..11817) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 11852..11871 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(11931..11947) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(11931..11947) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(11944..11966) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 11955..11971 /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(11979..12009) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 12024..12045 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(12162..12179) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(12333..12921) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(12413..12432) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(13092..13952) /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 13715..13734 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(13953..14057) /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 gctagctgca aagatggata aagttttaaa cagagaggaa 2641 tctttgcagc taatggacct tctaggtctt gaaaggagtg ggaattggct ccggtgcccg 2701 tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa 2761 ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg 2821 gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa 2881 cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt gtggttcccg 2941 cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt ccacctggct 3001 gcagtacgtg attcttgatc ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc 3061 ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg 3121 ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg ataagtctct 3181 agccatttaa aatttttgat gacctgctgc gacgcttttt ttctggcaag atagtcttgt 3241 aaatgcgggc caagatctgc acactggtat ttcggttttt ggggccgcgg gcggcgacgg 3301 ggcccgtgcg tcccagcgca catgttcggc gaggcggggc ctgcgagcgc ggccaccgag 3361 aatcggacgg gggtagtctc aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc 3421 gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga 3481 aagatggccg cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg 3541 agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc 3601 ttcatgtgac tccacggagt accgggcgcc gtccaggcac ctcgattagt tctcgagctt 3661 ttggagtacg tcgtctttag gttgggggga ggggttttat gcgatggagt ttccccacac 3721 tgagtgggtg gagactgaag ttaggccagc ttggcacttg atgtaattct ccttggaatt 3781 tgcccttttt gagtttggat cttggttcat tctcaagcct cagacagtgg ttcaaagttt 3841 ttttcttcca tttcaggtgt cgtgacgtac ggccaccatg aaaaggccgg cggccacgaa 3901 aaaggccggc caggcaaaaa agaaaaagga caagaagtac agcatcggcc tggccatcgg 3961 caccaactct gtgggctggg ccgtgatcac cgacgagtac aaggtgccca gcaagaaatt 4021 caaggtgctg ggcaacaccg accggcacag catcaagaag aacctgatcg gagccctgct 4081 gttcgacagc ggcgaaacag ccgaggccac ccggctgaag agaaccgcca gaagaagata 4141 caccagacgg aagaaccgga tctgctatct gcaagagatc ttcagcaacg agatggccaa 4201 ggtggacgac agcttcttcc acagactgga agagtccttc ctggtggaag aggataagaa 4261 gcacgagcgg caccccatct tcggcaacat cgtggacgag gtggcctacc acgagaagta 4321 ccccaccatc taccacctga gaaagaaact ggtggacagc accgacaagg ccgacctgcg 4381 gctgatctat ctggccctgg cccacatgat caagttccgg ggccacttcc tgatcgaggg 4441 cgacctgaac cccgacaaca gcgacgtgga caagctgttc atccagctgg tgcagaccta 4501 caaccagctg ttcgaggaaa accccatcaa cgccagcggc gtggacgcca aggccatcct 4561 gtctgccaga ctgagcaaga gcagacggct ggaaaatctg atcgcccagc tgcccggcga 4621 gaagaagaat ggcctgttcg gcaacctgat tgccctgagc ctgggcctga cccccaactt 4681 caagagcaac ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg acacctacga 4741 cgacgacctg gacaacctgc tggcccagat cggcgaccag tacgccgacc tgtttctggc 4801 cgccaagaac ctgtccgacg ccatcctgct gagcgacatc ctgagagtga acaccgagat 4861 caccaaggcc cccctgagcg cctctatgat caagagatac gacgagcacc accaggacct 4921 gaccctgctg aaagctctcg tgcggcagca gctgcctgag aagtacaaag agattttctt 4981 cgaccagagc aagaacggct acgccggcta cattgacggc ggagccagcc aggaagagtt 5041 ctacaagttc atcaagccca tcctggaaaa gatggacggc accgaggaac tgctcgtgaa 5101 gctgaacaga gaggacctgc tgcggaagca gcggaccttc gacaacggca gcatccccca 5161 ccagatccac ctgggagagc tgcacgccat tctgcggcgg caggaagatt tttacccatt 5221 cctgaaggac aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc cctactacgt 5281 gggccctctg gccaggggaa acagcagatt cgcctggatg accagaaaga gcgaggaaac 5341 catcaccccc tggaacttcg aggaagtggt ggacaagggc gcttccgccc agagcttcat 5401 cgagcggatg accaacttcg ataagaacct gcccaacgag aaggtgctgc ccaagcacag 5461 cctgctgtac gagtacttca ccgtgtataa cgagctgacc aaagtgaaat acgtgaccga 5521 gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg tggacctgct 5581 gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa gaggactact tcaagaaaat 5641 cgagtgcttc gactccgtgg aaatctccgg cgtggaagat cggttcaacg cctccctggg 5701 cacataccac gatctgctga aaattatcaa ggacaaggac ttcctggaca atgaggaaaa 5761 cgaggacatt ctggaagata tcgtgctgac cctgacactg tttgaggaca gagagatgat 5821 cgaggaacgg ctgaaaacct atgcccacct gttcgacgac aaagtgatga agcagctgaa 5881 gcggcggaga tacaccggct ggggcaggct gagccggaag ctgatcaacg gcatccggga 5941 caagcagtcc ggcaagacaa tcctggattt cctgaagtcc gacggcttcg ccaacagaaa 6001 cttcatgcag ctgatccacg acgacagcct gacctttaaa gaggacatcc agaaagccca 6061 ggtgtccggc cagggcgata gcctgcacga gcacattgcc aatctggccg gcagccccgc 6121 cattaagaag ggcatcctgc agacagtgaa ggtggtggac gagctcgtga aagtgatggg 6181 ccggcacaag cccgagaaca tcgtgatcga aatggccaga gagaaccaga ccacccagaa 6241 gggacagaag aacagccgcg agagaatgaa gcggatcgaa gagggcatca aagagctggg 6301 cagccagatc ctgaaagaac accccgtgga aaacacccag ctgcagaacg agaagctgta 6361 cctgtactac ctgcagaatg ggcgggatat gtacgtggac caggaactgg acatcaaccg 6421 gctgtccgac tacgatgtgg accacatcgt gcctcagagc tttctgaagg acgactccat 6481 cgacaacaag gtgctgacca gaagcgacaa ggcccggggc aagagcgaca acgtgccctc 6541 cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg ccaagctgat 6601 tacccagaga aagttcgaca atctgaccaa ggccgagaga ggcggcctga gcgaactgga 6661 taaggccggc ttcatcaaga gacagctggt ggaaacccgg cagatcacaa agcacgtggc 6721 acagatcctg gactcccgga tgaacactaa gtacgacgag aatgacaagc tgatccggga 6781 agtgaaagtg atcaccctga agtccaagct ggtgtccgat ttccggaagg atttccagtt 6841 ttacaaagtg cgcgagatca acaactacca ccacgcccac gacgcctacc tgaacgccgt 6901 cgtgggaacc gccctgatca aaaagtaccc taagctggaa agcgagttcg tgtacggcga 6961 ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc 7021 taccgccaag tacttcttct acagcaacat catgaacttt ttcaagaccg agattaccct 7081 ggccaacggc gagatccgga agcggcctct gatcgagaca aacggcgaaa ccggggagat 7141 cgtgtgggat aagggccggg attttgccac cgtgcggaaa gtgctgagca tgccccaagt 7201 gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc agcaaagagt ctatcctgcc 7261 caagaggaac agcgataagc tgatcgccag aaagaaggac tgggacccta agaagtacgg 7321 cggcttcgac agccccaccg tggcctattc tgtgctggtg gtggccaaag tggaaaaggg 7381 caagtccaag aaactgaaga gtgtgaaaga gctgctgggg atcaccatca tggaaagaag 7441 cagcttcgag aagaatccca tcgactttct ggaagccaag ggctacaaag aagtgaaaaa 7501 ggacctgatc atcaagctgc ctaagtactc cctgttcgag ctggaaaacg gccggaagag 7561 aatgctggcc tctgccggcg aactgcagaa gggaaacgaa ctggccctgc cctccaaata 7621 tgtgaacttc ctgtacctgg ccagccacta tgagaagctg aagggctccc ccgaggataa 7681 tgagcagaaa cagctgtttg tggaacagca caagcactac ctggacgaga tcatcgagca 7741 gatcagcgag ttctccaaga gagtgatcct ggccgacgct aatctggaca aagtgctgtc 7801 cgcctacaac aagcaccggg ataagcccat cagagagcag gccgagaata tcatccacct 7861 gtttaccctg accaatctgg gagcccctgc cgccttcaag tactttgaca ccaccatcga 7921 ccggaagagg tacaccagca ccaaagaggt gctggacgcc accctgatcc accagagcat 7981 caccggcctg tacgagacac ggatcgacct gtctcagctg ggaggcgaca gcgctggagg 8041 aggtggaagc ggaggaggag gaagcggagg aggaggtagc ggacctaaga aaaagaggaa 8101 ggtggcggcc gctggatccg gacgggctga cgcattggac gattttgatc tggatatgct 8161 gggaagtgac gccctcgatg attttgacct tgacatgctt ggttcggatg cccttgatga 8221 ctttgacctc gacatgctcg gcagtgacgc ccttgatgat ttcgacctgg acatgctgat 8281 taactgtaca ggcagtggag agggcagagg aagtctgcta acatgcggtg acgtcgagga 8341 gaatcctggc ccaatggcca agcctttgtc tcaagaagaa tccaccctca ttgaaagagc 8401 aacggctaca atcaacagca tccccatctc tgaagactac agcgtcgcca gcgcagctct 8461 ctctagcgac ggccgcatct tcactggtgt caatgtatat cattttactg ggggaccttg 8521 tgcagaactc gtggtgctgg gcactgctgc tgctgcggca gctggcaacc tgacttgtat 8581 cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc tgcggacggt gccgacaggt 8641 gcttctcgat ctgcatcctg ggatcaaagc catagtgaag gacagtgatg gacagccgac 8701 ggcagttggg attcgtgaat tgctgccctc tggttatgtg tgggagggct aagaattcga 8761 tatcaagctt atcggtaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat 8821 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca 8881 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc 8941 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc 9001 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt 9061 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg 9121 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc 9181 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta 9241 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg 9301 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc 9361 cccgcatcga taccgtcgac ctcgagacct agaaaaacat ggagcaatca caagtagcaa 9421 tacagcagct accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg 9481 ttttccagtc acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct 9541 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca 9601 agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt ggcagaacta 9661 cacaccaggg ccagggatca gatatccact gacctttgga tggtgctaca agctagtacc 9721 agttgagcaa gagaaggtag aagaagccaa tgaaggagag aacacccgct tgttacaccc 9781 tgtgagcctg catgggatgg atgacccgga gagagaagta ttagagtgga ggtttgacag 9841 ccgcctagca tttcatcaca tggcccgaga gctgcatccg gactgtactg ggtctctctg 9901 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc 9961 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg 10021 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gggcccgttt 10081 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct 10141 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg 10201 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc 10261 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct 10321 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct 10381 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg 10441 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg 10501 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac 10561 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct 10621 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt 10681 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt 10741 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt 10801 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag 10861 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc 10921 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc 10981 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg 11041 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca 11101 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg 11161 tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc 11221 ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt 11281 ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg 11341 ttctcccggg acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg 11401 ttcatcagcg cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg 11461 cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac 11521 gcctccgggc cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg 11581 cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtgcta 11641 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg 11701 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc 11761 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca 11821 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct 11881 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg 11941 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata 12001 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca 12061 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc 12121 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg 12181 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta 12241 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc 12301 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag 12361 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac 12421 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc 12481 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt 12541 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc 12601 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga 12661 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta 12721 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta 12781 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga 12841 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg 12901 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag 12961 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc 13021 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact 13081 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt 13141 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta 13201 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta 13261 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc 13321 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat 13381 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt 13441 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg 13501 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca 13561 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta 13621 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg 13681 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact 13741 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg 13801 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt 13861 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga 13921 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc 13981 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa 14041 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgac //