LOCUS Exported 14823 bp ds-DNA linear SYN 21-MAY-2021 DEFINITION dCas9 fused to human DNMT3A catalytic domain. ACCESSION . VERSION . KEYWORDS pLenti-EF1a-SPdCas9-DNMT3A-2A-Blast SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 14823) AUTHORS Lin L, Liu Y, Xu F, Huang J, Daugaard TF, Petersen TS, Hansen B, Ye L, Zhou Q, Fang F, Yang L, Li S, Floe L, Jensen KT, Shrock E, Chen F, Yang H, Wang J, Liu X, Xu X, Bolund L, Nielsen AL, Luo Y TITLE Genome-wide determination of on-target and off-target characteristics for RNA-guided DNA methylation by dCas9 methyltransferases. JOURNAL Gigascience. 2018 Mar 1;7(3):1-19. doi: 10.1093/gigascience/giy011. PUBMED 29635374 REFERENCE 2 (bases 1 to 14823) AUTHORS . TITLE Direct Submission JOURNAL Exported May 21, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..14823 /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 9038..9091 /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 9092..9490 /codon_start=1 /gene="Aspergillus terreus BSD" /product="blasticidin S deaminase" /label=BSD /note="confers resistance to blasticidin" /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" misc_feature 9515..10103 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(9568..9588) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(9986..9997) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(10106..10122) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(10107..10123) /label=pBluescriptKS /note="For pBluescript vector" LTR 10628..10808 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(10834..10851) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 10840..11064 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 11110..11538 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(11197..11216) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 11407..11428 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(11547..11567) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 11552..11881 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 11732..11867 /label=SV40 ori /note="SV40 origin of replication" primer_bind 11794..11813 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 11929..11976 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 11995..12369 /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 12499..12620 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(12536..12555) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 12590..12609 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(12669..12685) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(12669..12685) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(12682..12704) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 12693..12709 /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(12717..12747) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 12762..12783 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(12900..12917) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(13071..13659) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(13151..13170) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(13830..14690) /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 14453..14472 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(14691..14795) /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 gctggatcca accacgacca ggaatttgac cctccaaagg tttacccacc 8161 tgtcccagct gagaagagga agcccatccg ggtgctgtct ctctttgatg gaatcgctac 8221 agggctcctg gtgctgaagg acttgggcat tcaggtggac cgctacattg cctcggaggt 8281 gtgtgaggac tccatcacgg tgggcatggt gcggcaccag gggaagatca tgtacgtcgg 8341 ggacgtccgc agcgtcacac agaagcatat ccaggagtgg ggcccattcg atctggtgat 8401 tgggggcagt ccctgcaatg acctctccat cgtcaaccct gctcgcaagg gcctctacga 8461 gggcactggc cggctcttct ttgagttcta ccgcctcctg catgatgcgc ggcccaagga 8521 gggagatgat cgccccttct tctggctctt tgagaatgtg gtggccatgg gcgttagtga 8581 caagagggac atctcgcgat ttctcgagtc caaccctgtg atgattgatg ccaaagaagt 8641 gtcagctgca cacagggccc gctacttctg gggtaacctt cccggtatga acaggccgtt 8701 ggcatccact gtgaatgata agctggagct gcaggagtgt ctggagcatg gcaggatagc 8761 caagttcagc aaagtgagga ccattactac gaggtcaaac tccataaagc agggcaaaga 8821 ccagcatttt cctgtcttca tgaatgagaa agaggacatc ttatggtgca ctgaaatgga 8881 aagggtattt ggtttcccag tccactatac tgacgtctcc aacatgagcc gcttggcgag 8941 gcagagactg ctgggccggt catggagcgt gccagtcatc cgccacctct tcgctccgct 9001 gaaggagtat tttgcgtgtg tgtgtacagg cagtggagag ggcagaggaa gtctgctaac 9061 atgcggtgac gtcgaggaga atcctggccc aatggccaag cctttgtctc aagaagaatc 9121 caccctcatt gaaagagcaa cggctacaat caacagcatc cccatctctg aagactacag 9181 cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc actggtgtca atgtatatca 9241 ttttactggg ggaccttgtg cagaactcgt ggtgctgggc actgctgctg ctgcggcagc 9301 tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac aggggcatct tgagcccctg 9361 cggacggtgc cgacaggtgc ttctcgatct gcatcctggg atcaaagcca tagtgaagga 9421 cagtgatgga cagccgacgg cagttgggat tcgtgaattg ctgccctctg gttatgtgtg 9481 ggagggctaa gaattcgata tcaagcttat cggtaatcaa cctctggatt acaaaatttg 9541 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc 9601 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta 9661 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt 9721 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca 9781 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc 9841 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt 9901 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg 9961 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg 10021 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat 10081 ctccctttgg gccgcctccc cgcatcgata ccgtcgacct cgagacctag aaaaacatgg 10141 agcaatcaca agtagcaata cagcagctac caatgctgat tgtgcctggc tagaagcaca 10201 agaggaggag gaggtgggtt ttccagtcac acctcaggta cctttaagac caatgactta 10261 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat 10321 tcactcccaa cgaagacaag atatccttga tctgtggatc taccacacac aaggctactt 10381 ccctgattgg cagaactaca caccagggcc agggatcaga tatccactga cctttggatg 10441 gtgctacaag ctagtaccag ttgagcaaga gaaggtagaa gaagccaatg aaggagagaa 10501 cacccgcttg ttacaccctg tgagcctgca tgggatggat gacccggaga gagaagtatt 10561 agagtggagg tttgacagcc gcctagcatt tcatcacatg gcccgagagc tgcatccgga 10621 ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg 10681 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg 10741 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat 10801 ctctagcagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc 10861 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg 10921 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc 10981 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg 11041 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg 11101 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca 11161 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct 11221 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt 11281 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac 11341 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct 11401 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt 11461 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac 11521 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc 11581 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg 11641 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc 11701 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc 11761 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc 11821 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa 11881 aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtgt tgacaattaa 11941 tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta aaccatggcc 12001 aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc 12061 tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc 12121 cgggacgacg tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc ggacaacacc 12181 ctggcctggg tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg 12241 tccacgaact tccgggacgc ctccgggccg gccatgaccg agatcggcga gcagccgtgg 12301 gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag 12361 caggactgac acgtgctacg agatttcgat tccaccgccg ccttctatga aaggttgggc 12421 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg 12481 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat 12541 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc 12601 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg 12661 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac 12721 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca 12781 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat 12841 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc 12901 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca 12961 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca 13021 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg 13081 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg 13141 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt 13201 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt 13261 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc 13321 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt 13381 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt 13441 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc 13501 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa 13561 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt 13621 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct 13681 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta 13741 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa 13801 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc 13861 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact 13921 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc 13981 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt 14041 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta 14101 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg 14161 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt 14221 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc 14281 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt 14341 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc 14401 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc 14461 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa 14521 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac 14581 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa 14641 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt 14701 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa 14761 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct 14821 gac //