出品公司: | ORIGENE |
---|---|
载体名称: | pCas-Guide-EF1a-CD4 |
质粒类型: | CRISPR/Cas9载体;基因敲除载体;哺乳动物细胞载体 |
高拷贝/低拷贝: | 高拷贝 |
插入位点: | 限制性内切酶,多克隆位点 |
启动子: | U6 |
载体大小: | 11006 bp |
5' 测序引物及序列: | 5’ gatcgATGGGAGGTGGTATGGGAGGg 3’ |
3' 测序引物及序列: | 5’ aaaacCCTCCCATACCACCTCCCATc 3’ |
载体标签: | Myc(C-ter) |
载体抗性: | 氨苄青霉素 |
筛选标记: | -- |
克隆菌株: | Stbl3 |
宿主细胞(系): | 哺乳动物细胞 |
备注: | -- |
产品目录号: | GE100022 |
稳定性: | 瞬表达 |
组成型/诱导型: | 组成型 |
病毒/非病毒: | 非病毒 |
买家导航
pCas-Guide-EF1a-CD4载体质粒基本信息
pCas-Guide-EF1a-CD4质粒图谱载体图谱和pCas-Guide-EF1a-CD4载体序列质粒序列多克隆位点信息
pCas-Guide-EF1a-CD4质粒载体简介
pCas-Guide-EF1a-CD4载体特征
All-in-one vector
Expresses human codon-optimized Cas9, C-terminal Myc-DDK tagged
BamH I and BsmB I sites are used for genomic target sequence cloning
Expresses gRNA containing the inserted target sequence and the gRNA scaffold
EF1a promoter driven CD4 to enrich transfected cells
CF3 primer (GE100008) can be used to sequence the target sequence after cloning
pCas-Guide-EF1a-CD4质粒序列载体序列
LOCUS Exported File 11006 bp ds-DNA circular SYN 23-七月-2016 DEFINITION Mammalian vector for co-expressing a guide RNA (gRNA) together with Cas9 and CD4, for genome editing using the S. pyogenes CRISPR/Cas9 system. ACCESSION . VERSION . KEYWORDS pCas-Guide-EF1a-CD4 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 11006) AUTHORS OriGene TITLE Direct Submission JOURNAL Exported 2016-07-23 from SnapGene Viewer 1.5.3 http://www.snapgene.com FEATURES Location/Qualifiers source 1..11006 /organism="synthetic DNA construct" /lab_host="Mammalian Cells" /mol_type="other DNA" promoter 90..330 /note="U6" /note="RNA polymerase III promoter for human U6 snRNA" /note="color: #000000; direction: RIGHT" misc_RNA 384..459 /note="gRNA scaffold" /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" /note="color: #00ccff; direction: RIGHT" terminator 460..465 /note="polIII terminator" /note="RNA polymerase III transcription terminator" /note="color: #ffffff" promoter 480..1658 /note="EF-1-alpha promoter" /note="strong constitutive promoter for human elongation factor EF-1-alpha" /note="color: #000000; direction: RIGHT" intron 711..1649 /note="EF-1-alpha intron A" /note="intron upstream of the start codon of human EF-1-alpha" /note="color: #------" CDS 1679..2950 /codon_start=1 /product="truncated human CD4 cell surface glycoprotein lacking the cytoplasmic tail" /note="CD4 (truncated)" /note="This feature has 2 segments: ???1:?1679?..?1753?/?#ff6600?/?signal?peptide ???2:?1754?..?2950?/?#ff6600 Cleavage site after base 1753" /translation="MNRGVPFRHLLLVLQLALLPAATQGKKVVLGKKGDTVELTCTASQ KKSIQFHWKNSNQIKILGNQGSFLTKGPSKLNDRADSRRSLWDQGNFPLIIKNLKIEDS DTYICEVEDQKEEVQLLVFGLTANSDTHLLQGQSLTLTLESPPGSSPSVQCRSPRGKNI QGGKTLSVSQLELQDSGTWTCTVLQNQKKVEFKIDIVVLAFQKASSIVYKKEGEQVEFS FPLAFTVEKLTGSGELWWQAERASSSKSWITFDLKNKEVSVKRVTQDPKLQMGKKLPLH LTLPQALPQYAGSGNLTLALEAKTGKLHQEVNLVVMRATQLQKNLTCEVWGPTSPKLML SLKLENKEAKVSKREKAVWVLNPEAGMWQCLLSDSGQVLLESNIKVLPTWSTPVQPMAL IVLGGVAGLLLFIGLGIFFCVRCRH" enhancer 3560..3863 /note="CMV enhancer" /note="human cytomegalovirus immediate early enhancer" /note="color: #000000" promoter 3864..4067 /note="CMV promoter" /note="human cytomegalovirus (CMV) immediate early promoter" /note="color: #000000; direction: RIGHT" promoter 4093..4111 /note="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" /note="color: #ffffff; direction: RIGHT" CDS 4170..8273 /codon_start=1 /locus_tag=" " /product="Cas9 (Csn1) endonuclease from the?Streptococcus pyogenes?Type II CRISPR/Cas system" /note="Cas9" /note="generates RNA-guided double strand breaks in DNA" /note="color: #993366" /protein_id=" " /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 8274..8294 /codon_start=1 /locus_tag=" " /product="nuclear localization signal of SV40 large T antigen" /note="SV40 NLS" /note=" " /note="color: #993366" /protein_id=" " /translation="PKKKRKV" CDS 8301..8321 /codon_start=1 /locus_tag=" " /product="nuclear localization signal of SV40 large T antigen" /note="SV40 NLS" /note=" " /note="color: #993366" /protein_id=" " /translation="PKKKRKV" CDS 8343..8372 /codon_start=1 /product="Myc (human c-Myc oncogene) epitope tag" /note="Myc" /note="color: #cc99b2" /translation="EQKLISEEDL" CDS 8391..8414 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /note="FLAG" /note="color: #cc99b2" /translation="DYKDDDDK" polyA_signal 8461..9083 /note="hGH poly(A) signal" /note="human growth hormone polyadenylation signal" /note="color: #a6acb3" rep_origin complement(9204..9792) /direction=LEFT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /note="color: #ffff00" CDS complement(9967..10827) /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /note="This feature has 2 segments: ???1:??9967?..?10758?/?#ccffcc ???2:?10759?..?10827?/?#ccffcc?/?signal?sequence Cleavage site after base 10758" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" ORIGIN 1 gaattcccca gtggaaagac gcgcaggcaa aacgcaccac gtgacggagc gtgaccgcgc 61 gccgagcgcg cgccaaggtc gggcaggaag agggcctatt tcccatgatt ccttcatatt 121 tgcatatacg atacaaggct gttagagaga taattagaat taatttgact gtaaacacaa 181 agatattagt acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt 241 taaaattatg ttttaaaatg gactatcata tgcttaccgt aacttgaaag tatttcgatt 301 tcttgggttt atatatcttg tggaaaggac gcgggatcca ctggaccagg cagcagcgtc 361 agaagacttt tttggaacgt ctcgttttag agctagaaat agcaagttaa aataaggcta 421 gtccgttatc aacttgaaaa agtggcaccg agtcggtgct ttttttggtg tacacgtgag 481 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg 541 gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg 601 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag 661 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg 721 tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta 781 cttccacctg gctgcagtac gtgattcttg atcccgagct tcgggttgga agtgggtggg 841 agagttcgag gccttgcgct taaggagccc cttcgcctcg tgcttgagtt gaggcctggc 901 ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt 961 tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc 1021 aagatagtct tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt tttggggccg 1081 cgggcggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag 1141 cgcggccacc gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg 1201 gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag 1261 ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc agggagctca aaatggagga 1321 cgcggcgctc gggagagcgg gcgggtgagt cacccacaca aaggaaaagg gcctttccgt 1381 cctcagccgt cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt 1441 agttctcgag cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg 1501 agtttcccca cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat 1561 tctccttgga atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag 1621 tggttcaaag tttttttctt ccatttcagg tgtcgtgact atagggcggc cggacgtgat 1681 gaaccgggga gtccctttta ggcacttgct tctggtgctg caactggcgc tcctcccagc 1741 agccactcag ggaaagaaag tggtgctggg caaaaaaggg gatacagtgg aactgacctg 1801 tacagcttcc cagaagaaga gcatacaatt ccactggaaa aactccaacc agataaagat 1861 tctgggaaat cagggctcct tcttaactaa aggtccatcc aagctgaatg atcgcgctga 1921 ctcaagaaga agcctttggg accaaggaaa ctttcccctg atcatcaaga atcttaagat 1981 agaagactca gatacttaca tctgtgaagt ggaggaccag aaggaggagg tgcaattgct 2041 agtgttcgga ttgactgcca actctgacac ccacctgctt caggggcaga gcctgaccct 2101 gaccttggag agcccccctg gtagtagccc ctcagtgcaa tgtaggagtc caaggggtaa 2161 aaacatacag ggggggaaga ccctctccgt gtctcagctg gagctccagg atagtggcac 2221 ctggacatgc actgtcttgc agaaccagaa gaaggtggag ttcaaaatag acatcgtggt 2281 gctagctttc cagaaggcct ccagcatagt ctataagaaa gagggggaac aggtggagtt 2341 ctccttccca ctcgccttta cagttgaaaa gctgacgggc agtggcgagc tgtggtggca 2401 ggcggagagg gcttcctcct ccaagtcttg gatcaccttt gacctgaaga acaaggaagt 2461 gtctgtaaaa cgggttaccc aggaccctaa gctccagatg ggcaagaagc tcccgctcca 2521 cctcaccctg ccccaggcct tgcctcagta tgctggctct ggaaacctca ccctggccct 2581 tgaagcgaaa acaggaaagt tgcatcagga agtgaacctg gtggtgatga gagccactca 2641 gctccagaaa aatttgacct gtgaggtgtg gggacccacc tcccctaagc tgatgctgag 2701 cttgaaactg gagaacaagg aggcaaaggt ctcgaagcgg gagaaggcgg tgtgggtgct 2761 gaaccctgag gcggggatgt ggcagtgtct gctgagtgac tcgggacagg tcctgctgga 2821 atccaacatc aaggttctgc ccacatggtc caccccggtg cagccaatgg ccctgattgt 2881 gctggggggc gtcgccggcc tcctgctttt cattgggcta ggcatcttct tctgtgtcag 2941 gtgccggcac accggttagg atatcagatc cccgggatgc agaaattgat gatctattaa 3001 acaataaaga tgtccactaa aatggaagtt tttcctgtca tactttgtta agaagggtga 3061 gaacagagta cctacatttt gaatggaagg attggagcta cgggggtggg ggtggggtgg 3121 gattagataa atgcctgctc tttactgaag gctctttact attgctttat gataatgttt 3181 catagttgga tatcataatt taaacaagca aaaccaaatt aagggccagc tcattcctcc 3241 cactcatgat ctatagatct atagatctct cgtgggatca ttgtttttct cttgattccc 3301 actttgtggt tctaagtact gtggtttcca aatgtgtcag tttcatagcc tgaagaacga 3361 gatcagcagc ctctgttcca catacacttc attctcagta ttgttttgcc aagttctaat 3421 tccatcagaa gctggtcgag atccggaacc cttaatataa cttcgtataa tgtatgctat 3481 acgaagttat taggtccact agttattaat agtaatcaat tacggggtca ttagttcata 3541 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 3601 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag 3661 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac 3721 atcaagtgta tcatatgcca agtccgcccc ctattgacgt caatgacggt aaatggcccg 3781 cctggcatta tgcccagtac atgaccttac gggactttcc tacttggcag tacatctacg 3841 tattagtcat cgctattacc atggtgatgc ggttttggca gtacaccaat gggcgtggat 3901 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt 3961 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa taaccccgcc ccgttgacgc 4021 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc 4081 gtcagaattt tgtaatacga ctcactatag ggcggccggg aattcgtcga ctggaaccgg 4141 taccgaggag atctgccgcc gcgatcgcca tggataagaa atactcaata ggactggata 4201 ttggcacaaa tagcgtcgga tgggctgtga tcactgatga atataaggtt ccttctaaaa 4261 agttcaaggt tctgggaaat acagaccgcc acagtatcaa aaaaaatctt ataggggctc 4321 ttctgtttga cagtggagag acagccgaag ctactagact caaacggaca gctaggagaa 4381 ggtatacaag acggaagaat aggatttgtt atctccagga gattttttca aatgagatgg 4441 ccaaagtgga tgatagtttc tttcatagac ttgaagagtc ttttttggtg gaagaagaca 4501 agaagcatga aagacatcct atttttggaa atatagtgga tgaagttgct tatcacgaga 4561 aatatccaac tatctatcat ctgagaaaaa aattggtgga ttctactgat aaagccgatt 4621 tgcgcctgat ctatttggcc ctggcccaca tgattaagtt tagaggtcat tttttgattg 4681 agggcgatct gaatcctgat aatagtgatg tggacaaact gtttatccag ttggtgcaaa 4741 cctacaatca actgtttgaa gaaaacccta ttaacgcaag tggagtggat gctaaagcca 4801 ttctttctgc aagattgagt aaatcaagaa gactggaaaa tctcattgct cagctccccg 4861 gtgagaagaa aaatggcctg tttgggaatc tcattgcttt gtcattgggt ttgaccccta 4921 attttaaatc aaattttgat ttggcagaag atgctaaact ccagctttca aaagatactt 4981 acgatgatga tctggataat ctgttggctc aaattggaga tcaatatgct gatttgtttt 5041 tggcagctaa gaatctgtca gatgctattc tgctttcaga catcctgaga gtgaatactg 5101 aaataactaa ggctcccctg tcagcttcaa tgattaaacg ctacgatgaa catcatcaag 5161 acttgactct tctgaaagcc ctggttagac aacaacttcc agaaaagtat aaagaaatct 5221 tttttgatca atcaaaaaac ggatatgcag gttatattga tggcggcgca agccaagaag 5281 aattttataa atttatcaaa ccaattctgg aaaaaatgga tggtactgag gaactgttgg 5341 tgaaactgaa tagagaagat ttgctgcgca agcaacggac ctttgacaac ggctctattc 5401 cccatcaaat tcacttgggt gagctgcatg ctattttgag aagacaagaa gacttttatc 5461 catttctgaa agacaataga gagaagattg aaaaaatctt gacttttagg attccttatt 5521 atgttggtcc attggccaga ggcaatagta ggtttgcatg gatgactcgg aagtctgaag 5581 aaacaattac cccatggaat tttgaagaag ttgtcgataa aggtgcttca gctcaatcat 5641 ttattgaacg catgacaaac tttgataaaa atcttccaaa tgaaaaagtg ctgccaaaac 5701 atagtttgct ttatgagtat tttaccgttt ataacgaatt gacaaaggtc aaatatgtta 5761 ctgaaggaat gagaaaacca gcatttcttt caggtgaaca gaagaaagcc attgttgatc 5821 tgctcttcaa aacaaatagg aaagtgaccg ttaagcaact gaaagaagat tatttcaaaa 5881 aaatagaatg ttttgatagt gttgaaattt caggagttga agatagattt aatgcttcac 5941 tgggtacata ccatgatttg ctgaaaatta ttaaagataa agattttttg gataatgaag 6001 aaaatgaaga catcctggag gatattgttc tgacattgac cctgtttgaa gatagggaga 6061 tgattgagga aagacttaaa acatacgctc acctctttga tgataaggtg atgaaacagc 6121 ttaaaagacg cagatatact ggttggggaa ggttgtccag aaaattgatt aatggtatta 6181 gggataagca atctggcaaa acaatactgg attttttgaa atcagatggt tttgccaatc 6241 gcaattttat gcagctcatc catgatgata gtttgacatt taaagaagac atccaaaaag 6301 cacaagtgtc tggacaaggc gatagtctgc atgaacatat tgcaaatctg gctggtagcc 6361 ctgctattaa aaaaggtatt ctccagactg tgaaagttgt tgatgaattg gtcaaagtga 6421 tggggcggca taagccagaa aatatcgtta ttgaaatggc aagagaaaat cagacaactc 6481 aaaagggcca gaaaaattcc agagagagga tgaaaagaat cgaagaaggt atcaaagaac 6541 tgggaagtca gattcttaaa gagcatcctg ttgaaaatac tcaattgcaa aatgaaaagc 6601 tctatctcta ttatctccaa aatggaagag atatgtatgt ggaccaagaa ctggatatta 6661 ataggctgag tgattatgat gtcgatcaca ttgttccaca aagtttcctt aaagacgatt 6721 caatagacaa taaggtcctg accaggtctg ataaaaatag aggtaaatcc gataacgttc 6781 caagtgaaga agtggtcaaa aagatgaaaa actattggag acaacttctg aacgccaagc 6841 tgatcactca aaggaagttt gataatctga ccaaagctga aagaggaggt ttgagtgaac 6901 ttgataaagc tggttttatc aaacgccaat tggttgaaac tcgccaaatc actaagcatg 6961 tggcacaaat tttggatagt cgcatgaata ctaaatacga tgaaaatgat aaacttatta 7021 gagaggttaa agtgattacc ctgaaatcta aactggtttc tgacttcaga aaagatttcc 7081 aattctataa agtgagagag attaacaatt accatcatgc ccatgatgcc tatctgaatg 7141 ccgtcgttgg aactgctttg attaagaaat atccaaaact tgaaagcgag tttgtctatg 7201 gtgattataa agtttatgat gttaggaaaa tgattgctaa gtctgagcaa gaaataggca 7261 aagcaaccgc aaagtatttc ttttactcta atatcatgaa cttcttcaaa acagaaatta 7321 cacttgcaaa tggagagatt cgcaaacgcc ctctgatcga aactaatggg gaaactggag 7381 aaattgtctg ggataaaggg agagattttg ccacagtgcg caaagtgttg tccatgcccc 7441 aagtcaatat cgtcaagaaa acagaagtgc agacaggcgg attctctaag gagtcaattc 7501 tgccaaaaag aaattccgac aagctgattg ctaggaaaaa agactgggac ccaaaaaaat 7561 atggtggttt tgatagtcca accgtggctt attcagtcct ggtggttgct aaggtggaaa 7621 aagggaaatc caagaagctg aaatccgtta aagagctgct ggggatcaca attatggaaa 7681 gaagttcctt tgaaaaaaat cccattgact ttctggaagc taaaggatat aaggaagtta 7741 aaaaagacct gatcattaaa ctgcctaaat atagtctttt tgagctggaa aacggtagga 7801 aacggatgct ggctagtgcc ggagaactgc aaaaaggaaa tgagctggct ctgccaagca 7861 aatatgtgaa ttttctgtat ctggctagtc attatgaaaa gttgaagggt agtccagaag 7921 ataacgaaca aaaacaattg tttgtggagc agcataagca ttatctggat gagattattg 7981 agcaaatcag tgaattttct aagagagtta ttctggcaga tgccaatctg gataaagttc 8041 ttagtgcata taacaaacat agagacaaac caataagaga acaagcagaa aatatcattc 8101 atctgtttac cttgaccaat cttggagcac ccgctgcttt taaatacttt gatacaacaa 8161 ttgataggaa aagatatacc tctacaaaag aagttctgga tgccactctt atccatcaat 8221 ccatcactgg tctttatgaa acacgcattg atttgagtca gctgggaggt gaccccaaga 8281 aaaaacgcaa ggtggaagat cctaagaaaa agcggaaagt ggacacgcgt acgcggccgc 8341 tcgagcagaa actcatctca gaagaggatc tggcagcaaa tgatatcctg gattacaagg 8401 atgacgacga taaggtttaa acggccggcc gcggtcatag ctgtttcctg aacagatccc 8461 gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca 8521 gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc 8581 ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca 8641 acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg 8701 ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg 8761 ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagaggcgg 8821 ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct 8881 tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg 8941 attttaaaat aactatacca gcaggaggac gtccagacac agcataggct acctggccat 9001 gcccaaccgg tgggacattt gagttgcttg cttggcactg tcctctcatg cgttgggtcc 9061 actcagtaga tgcctgttga attgggtacg cggccagcgg cgagcggtat cagctcactc 9121 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtccgt 9181 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa 9241 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 9301 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 9361 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc 9421 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 9481 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 9541 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 9601 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc 9661 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 9721 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 9781 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgac 9841 gcgtaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 9901 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 9961 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 10021 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 10081 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 10141 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 10201 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 10261 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 10321 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 10381 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 10441 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 10501 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 10561 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 10621 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 10681 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 10741 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 10801 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 10861 tcagggttat tgtctcatga tgatattatt ttatcttgtg caatgtaaca tcagagattt 10921 tgagacacgg gccagagctg ccaggaaaca gctatgacca tgtaatacga ctcactatag 10981 gggatatcag ctggatggca gttaac //
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