LOCUS Exported 13012 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION 3rd generation lentiviral gRNA plasmid targeting human KAT5. ACCESSION . VERSION . KEYWORDS KAT5 sgRNA1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 13012) AUTHORS Kwan SY, Sheel A, Song CQ, Zhang XO, Jiang T, Dang H, Cao Y, Ozata DM, Mou H, Yin H, Weng Z, Wang XW, Xue W TITLE Depletion of TRRAP induces p53-independent senescence in liver cancer by downregulating mitotic genes. JOURNAL Hepatology. 2019 Jun 12. doi: 10.1002/hep.30807. PUBMED 31188495 REFERENCE 2 (bases 1 to 13012) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..13012 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..199 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 154..174 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 217..397 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 444..569 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1062..1295 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1480..1524 /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 1822..1939 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 1990..2230 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 1990..2010 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 2161..2180 /label=LKO.1 5' /note="Human U6 promoter, forward primer" misc_RNA 2259..2334 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" promoter 2396..2607 /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" regulatory 2626..2635 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2632..6735 /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL DATLIHQSITGLYETRIDLSQLGGD" CDS 6736..6783 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" CDS 6784..6807 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 6817..6873 /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 6874..7470 /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 7367..7387 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 7486..8074 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(7539..7559) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(7957..7968) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 8146..8379 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" primer_bind complement(8405..8422) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 8411..8635 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 8681..9109 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(8768..8787) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 8978..8999 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(9118..9138) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 9123..9452 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 9303..9438 /label=SV40 ori /note="SV40 origin of replication" primer_bind 9365..9384 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 9500..9547 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 9566..9940 /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 10070..10191 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(10107..10126) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 10161..10180 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(10240..10256) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(10240..10256) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(10253..10275) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 10264..10280 /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(10288..10318) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 10333..10354 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(10471..10488) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(10642..11230) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(10722..10741) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(11401..12261) /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 12024..12043 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(12262..12366) /gene="bla" /label=AmpR promoter primer_bind complement(12441..12460) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 12632..13011 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" ORIGIN 1 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc 61 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact 121 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt 181 gggaggtcta tataagcagc gcgttttgcc tgtactgggt ctctctggtt agaccagatc 241 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg 301 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc 361 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga 421 aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca 481 cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct 541 agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat 601 gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat 661 gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag 721 gctgtagaca aatactggga cagctacaac catcccttca gacaggatca gaagaactta 781 gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag 841 acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac 901 agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt 961 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca 1021 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg 1081 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc 1141 agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg 1201 caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg 1261 gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa 1321 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag 1381 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta 1441 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg 1501 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat 1561 ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta 1621 ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc 1681 ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac 1741 agagacagat ccattcgatt agtgaacgga tcggcactgc gtgcgccaat tctgcagaca 1801 aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg 1861 ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat 1921 tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccag tttggttaat 1981 taaggtaccg agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct 2041 gttagagaga taattagaat taatttgact gtaaacacaa agatattagt acaaaatacg 2101 tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg 2161 gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg 2221 tggaaaggac gaaacaccga ttgatggacg taagaacagt tttagagcta gaaatagcaa 2281 gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt 2341 gaattcgcta gctaggtctt gaaaggagtg ggaattggct ccggtgcccg tcagtgggca 2401 gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgatccggt 2461 gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt 2521 tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt 2581 cgcaacgggt ttgccgccag aacacaggac cggttctaga gcgctgccac catggacaag 2641 aagtacagca tcggcctgga catcggcacc aactctgtgg gctgggccgt gatcaccgac 2701 gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc 2761 aagaagaacc tgatcggagc cctgctgttc gacagcggcg aaacagccga ggccacccgg 2821 ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa 2881 gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag 2941 tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg 3001 gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg 3061 gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag 3121 ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag 3181 ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc 3241 agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa 3301 aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggaaa cctgattgcc 3361 ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa 3421 ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc 3481 gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc 3541 gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag 3601 agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg 3661 cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatt 3721 gacggcggag ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg 3781 gacggcaccg aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg 3841 accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg 3901 cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc 3961 ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc 4021 tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac 4081 aagggcgctt ccgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc 4141 aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtataacgag 4201 ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag 4261 cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag 4321 ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg 4381 gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac 4441 aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg 4501 acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc 4561 gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc 4621 cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg 4681 aagtccgacg gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc 4741 tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac 4801 attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg 4861 gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg 4921 gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg 4981 atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac 5041 acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac 5101 gtggaccagg aactggacat caaccggctg tccgactacg atgtggacca tatcgtgcct 5161 cagagctttc tgaaggacga ctccatcgac aacaaggtgc tgaccagaag cgacaagaac 5221 cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg 5281 cggcagctgc tgaacgccaa gctgattacc cagagaaagt tcgacaatct gaccaaggcc 5341 gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa 5401 acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac 5461 gacgagaatg acaagctgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg 5521 tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac 5581 gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag 5641 ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc 5701 aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg 5761 aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc 5821 gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg gccgggattt tgccaccgtg 5881 cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc 5941 ggcttcagca aagagtctat cctgcccaag aggaacagcg ataagctgat cgccagaaag 6001 aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg 6061 ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg 6121 ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa 6181 gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg 6241 ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga 6301 aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag 6361 aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaag 6421 cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc 6481 gacgctaatc tggacaaagt gctgtccgcc tacaacaagc accgggataa gcccatcaga 6541 gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc 6601 ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg 6661 gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct 6721 cagctgggag gcgacaagcg acctgccgcc acaaagaagg ctggacaggc taagaagaag 6781 aaagattaca aagacgatga cgataaggga tccggcgcaa caaacttctc tctgctgaaa 6841 caagccggag atgtcgaaga gaatcctgga ccgaccgagt acaagcccac ggtgcgcctc 6901 gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac 6961 cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa 7021 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc 7081 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag 7141 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa 7201 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggagtc 7261 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg 7321 gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc 7381 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc 7441 tggtgcatga cccgcaagcc cggtgcctga acgcgttaag tcgacaatca acctctggat 7501 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt 7561 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc 7621 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg 7681 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc 7741 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa 7801 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat 7861 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc 7921 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt 7981 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag 8041 acgagtcgga tctccctttg ggccgcctcc ccgcgtcgac tttaagacca atgacttaca 8101 aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc 8161 actcccaacg aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga 8221 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct 8281 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat 8341 ccctcagacc cttttagtca gtgtggaaaa tctctagcag ggcccgttta aacccgctga 8401 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct 8461 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca 8521 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag 8581 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct 8641 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca 8701 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta 8761 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt 8821 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac 8881 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt 8941 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga 9001 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg 9061 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga 9121 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 9181 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca 9241 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc 9301 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt 9361 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag 9421 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt 9481 tcggatctga tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata 9541 atacgacaag gtgaggaact aaaccatggc caagttgacc agtgccgttc cggtgctcac 9601 cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga 9661 cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc 9721 ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga 9781 cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc 9841 ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc 9901 cggcaactgc gtgcacttcg tggccgagga gcaggactga cacgtgctac gagatttcga 9961 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg 10021 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat 10081 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 10141 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg 10201 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg 10261 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc 10321 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt 10381 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg 10441 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt 10501 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc 10561 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 10621 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 10681 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 10741 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 10801 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag 10861 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 10921 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 10981 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 11041 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg 11101 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 11161 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 11221 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 11281 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 11341 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 11401 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 11461 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 11521 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 11581 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 11641 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 11701 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 11761 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 11821 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 11881 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 11941 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 12001 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 12061 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 12121 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 12181 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 12241 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 12301 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 12361 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat 12421 cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg 12481 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca 12541 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg 12601 cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag 12661 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt 12721 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg 12781 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat 12841 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct 12901 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg 12961 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat gg //