pCas-Guide-AAVS1

价格： ¥800 - 1200

品牌：Ybscience

货号：YB-10324

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载体基本信息

出品公司:	Ybscience
载体名称:	pCas-Guide-AAVS1
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:	8009 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:	--
稳定性:	瞬表达
组成型/诱导型:	组成型
病毒/非病毒:	非病毒

载体质粒图谱和多克隆位点信息

pCas-Guide-AAVS1载体图谱

载体简介

pCas-Guide-AAVS1载体特征Validated CRISPR vector targeting human AAVS1 siteAAVS1 target sequence: GTTAATGTGGCTCTGGTTCTCMV-driven Cas9 and U6-driven AAVS1 gRNA expression

载体序列

LOCUS       Exported File           8009 bp ds-DNA    circular SYN 23-七月-2016DEFINITION  Mammalian guide RNA (gRNA) vector for transgene integration at the             AAVS1 locus in the human genome. Use together with pAAVS1-puro-DNR.ACCESSION   .VERSION     .KEYWORDS    pCas-Guide-AAVS1SOURCE      synthetic DNA construct  ORGANISM  synthetic DNA constructREFERENCE   1  (bases 1 to 8009)  AUTHORS   OriGene  TITLE     Direct Submission  JOURNAL   Exported 2016-07-23 from SnapGene Viewer 1.5.3            http://www.snapgene.comFEATURES             Location/Qualifiers     source          1..8009                     /organism="synthetic DNA construct"                     /lab_host="Mammalian Cells"                     /mol_type="other DNA"     promoter        90..330                     /note="U6 promoter"                     /note="RNA polymerase III promoter for human U6 snRNA"                     /note="color: #000000; direction: RIGHT"     misc_RNA        340..359                     /note="AAVS1 target sequence"                     /note="color: #99cc00; direction: RIGHT"     misc_RNA        360..435                     /note="gRNA scaffold"                     /note="guide RNA scaffold for the Streptococcus pyogenes                      CRISPR/Cas9 system"                     /note="color: #00ccff; direction: RIGHT"     terminator      436..441                     /note="polIII terminator"                     /note="RNA polymerase III transcription terminator"                     /note="color: #ffffff"     enhancer        487..866                     /note="CMV enhancer"                     /note="human cytomegalovirus immediate early enhancer"                     /note="color: #000000"     promoter        867..1070                     /note="CMV promoter"                     /note="human cytomegalovirus (CMV) immediate early                      promoter"                     /note="color: #000000; direction: RIGHT"     CDS             1173..5276                     /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             5277..5297                     /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             5304..5324                     /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             5346..5375                     /codon_start=1                     /product="Myc (human c-Myc oncogene) epitope tag"                     /note="Myc"                     /note="color: #cc99b2"                     /translation="EQKLISEEDL"     CDS             5394..5417                     /codon_start=1                     /product="FLAG(R) epitope tag, followed by an enterokinase                      cleavage site"                     /note="FLAG"                     /note="color: #cc99b2"                     /translation="DYKDDDDK"     polyA_signal    5464..6086                     /note="hGH poly(A) signal"                     /note="human growth hormone polyadenylation signal"                     /note="color: #a6acb3"     rep_origin      complement(6207..6795)                     /direction=LEFT                     /note="ori"                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of                      replication"                     /note="color: #ffff00"     CDS             complement(6970..7830)                     /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:?6970?..?7761?/?#ccffcc                     ???2:?7762?..?7830?/?#ccffcc?/?signal?sequence                     Cleavage site after base 7761"                     /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 gcgggatcgg ttaatgtggc tctggttctg      361 ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg      421 gcaccgagtc ggtgcttttt ttggtgtaca tttatattgg ctcatgtcca atatgaccgc      481 catgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc      541 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac      601 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa      661 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag      721 tacatcaagt gtatcatatg ccaagtccgc cccctattga cgtcaatgac ggtaaatggc      781 ccgcctggca ttatgcccag tacatgacct tacgggactt tcctacttgg cagtacatct      841 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacacc aatgggcgtg      901 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt      961 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taataacccc gccccgttga     1021 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga     1081 accgtcagaa ttttgtaata cgactcacta tagggcggcc gggaattcgt cgactggaac     1141 cggtaccgag gagatctgcc gccgcgatcg ccatggataa gaaatactca ataggactgg     1201 atattggcac aaatagcgtc ggatgggctg tgatcactga tgaatataag gttccttcta     1261 aaaagttcaa ggttctggga aatacagacc gccacagtat caaaaaaaat cttatagggg     1321 ctcttctgtt tgacagtgga gagacagccg aagctactag actcaaacgg acagctagga     1381 gaaggtatac aagacggaag aataggattt gttatctcca ggagattttt tcaaatgaga     1441 tggccaaagt ggatgatagt ttctttcata gacttgaaga gtcttttttg gtggaagaag     1501 acaagaagca tgaaagacat cctatttttg gaaatatagt ggatgaagtt gcttatcacg     1561 agaaatatcc aactatctat catctgagaa aaaaattggt ggattctact gataaagccg     1621 atttgcgcct gatctatttg gccctggccc acatgattaa gtttagaggt cattttttga     1681 ttgagggcga tctgaatcct gataatagtg atgtggacaa actgtttatc cagttggtgc     1741 aaacctacaa tcaactgttt gaagaaaacc ctattaacgc aagtggagtg gatgctaaag     1801 ccattctttc tgcaagattg agtaaatcaa gaagactgga aaatctcatt gctcagctcc     1861 ccggtgagaa gaaaaatggc ctgtttggga atctcattgc tttgtcattg ggtttgaccc     1921 ctaattttaa atcaaatttt gatttggcag aagatgctaa actccagctt tcaaaagata     1981 cttacgatga tgatctggat aatctgttgg ctcaaattgg agatcaatat gctgatttgt     2041 ttttggcagc taagaatctg tcagatgcta ttctgctttc agacatcctg agagtgaata     2101 ctgaaataac taaggctccc ctgtcagctt caatgattaa acgctacgat gaacatcatc     2161 aagacttgac tcttctgaaa gccctggtta gacaacaact tccagaaaag tataaagaaa     2221 tcttttttga tcaatcaaaa aacggatatg caggttatat tgatggcggc gcaagccaag     2281 aagaatttta taaatttatc aaaccaattc tggaaaaaat ggatggtact gaggaactgt     2341 tggtgaaact gaatagagaa gatttgctgc gcaagcaacg gacctttgac aacggctcta     2401 ttccccatca aattcacttg ggtgagctgc atgctatttt gagaagacaa gaagactttt     2461 atccatttct gaaagacaat agagagaaga ttgaaaaaat cttgactttt aggattcctt     2521 attatgttgg tccattggcc agaggcaata gtaggtttgc atggatgact cggaagtctg     2581 aagaaacaat taccccatgg aattttgaag aagttgtcga taaaggtgct tcagctcaat     2641 catttattga acgcatgaca aactttgata aaaatcttcc aaatgaaaaa gtgctgccaa     2701 aacatagttt gctttatgag tattttaccg tttataacga attgacaaag gtcaaatatg     2761 ttactgaagg aatgagaaaa ccagcatttc tttcaggtga acagaagaaa gccattgttg     2821 atctgctctt caaaacaaat aggaaagtga ccgttaagca actgaaagaa gattatttca     2881 aaaaaataga atgttttgat agtgttgaaa tttcaggagt tgaagataga tttaatgctt     2941 cactgggtac ataccatgat ttgctgaaaa ttattaaaga taaagatttt ttggataatg     3001 aagaaaatga agacatcctg gaggatattg ttctgacatt gaccctgttt gaagataggg     3061 agatgattga ggaaagactt aaaacatacg ctcacctctt tgatgataag gtgatgaaac     3121 agcttaaaag acgcagatat actggttggg gaaggttgtc cagaaaattg attaatggta     3181 ttagggataa gcaatctggc aaaacaatac tggatttttt gaaatcagat ggttttgcca     3241 atcgcaattt tatgcagctc atccatgatg atagtttgac atttaaagaa gacatccaaa     3301 aagcacaagt gtctggacaa ggcgatagtc tgcatgaaca tattgcaaat ctggctggta     3361 gccctgctat taaaaaaggt attctccaga ctgtgaaagt tgttgatgaa ttggtcaaag     3421 tgatggggcg gcataagcca gaaaatatcg ttattgaaat ggcaagagaa aatcagacaa     3481 ctcaaaaggg ccagaaaaat tccagagaga ggatgaaaag aatcgaagaa ggtatcaaag     3541 aactgggaag tcagattctt aaagagcatc ctgttgaaaa tactcaattg caaaatgaaa     3601 agctctatct ctattatctc caaaatggaa gagatatgta tgtggaccaa gaactggata     3661 ttaataggct gagtgattat gatgtcgatc acattgttcc acaaagtttc cttaaagacg     3721 attcaataga caataaggtc ctgaccaggt ctgataaaaa tagaggtaaa tccgataacg     3781 ttccaagtga agaagtggtc aaaaagatga aaaactattg gagacaactt ctgaacgcca     3841 agctgatcac tcaaaggaag tttgataatc tgaccaaagc tgaaagagga ggtttgagtg     3901 aacttgataa agctggtttt atcaaacgcc aattggttga aactcgccaa atcactaagc     3961 atgtggcaca aattttggat agtcgcatga atactaaata cgatgaaaat gataaactta     4021 ttagagaggt taaagtgatt accctgaaat ctaaactggt ttctgacttc agaaaagatt     4081 tccaattcta taaagtgaga gagattaaca attaccatca tgcccatgat gcctatctga     4141 atgccgtcgt tggaactgct ttgattaaga aatatccaaa acttgaaagc gagtttgtct     4201 atggtgatta taaagtttat gatgttagga aaatgattgc taagtctgag caagaaatag     4261 gcaaagcaac cgcaaagtat ttcttttact ctaatatcat gaacttcttc aaaacagaaa     4321 ttacacttgc aaatggagag attcgcaaac gccctctgat cgaaactaat ggggaaactg     4381 gagaaattgt ctgggataaa gggagagatt ttgccacagt gcgcaaagtg ttgtccatgc     4441 cccaagtcaa tatcgtcaag aaaacagaag tgcagacagg cggattctct aaggagtcaa     4501 ttctgccaaa aagaaattcc gacaagctga ttgctaggaa aaaagactgg gacccaaaaa     4561 aatatggtgg ttttgatagt ccaaccgtgg cttattcagt cctggtggtt gctaaggtgg     4621 aaaaagggaa atccaagaag ctgaaatccg ttaaagagct gctggggatc acaattatgg     4681 aaagaagttc ctttgaaaaa aatcccattg actttctgga agctaaagga tataaggaag     4741 ttaaaaaaga cctgatcatt aaactgccta aatatagtct ttttgagctg gaaaacggta     4801 ggaaacggat gctggctagt gccggagaac tgcaaaaagg aaatgagctg gctctgccaa     4861 gcaaatatgt gaattttctg tatctggcta gtcattatga aaagttgaag ggtagtccag     4921 aagataacga acaaaaacaa ttgtttgtgg agcagcataa gcattatctg gatgagatta     4981 ttgagcaaat cagtgaattt tctaagagag ttattctggc agatgccaat ctggataaag     5041 ttcttagtgc atataacaaa catagagaca aaccaataag agaacaagca gaaaatatca     5101 ttcatctgtt taccttgacc aatcttggag cacccgctgc ttttaaatac tttgatacaa     5161 caattgatag gaaaagatat acctctacaa aagaagttct ggatgccact cttatccatc     5221 aatccatcac tggtctttat gaaacacgca ttgatttgag tcagctggga ggtgacccca     5281 agaaaaaacg caaggtggaa gatcctaaga aaaagcggaa agtggacacg cgtacgcggc     5341 cgctcgagca gaaactcatc tcagaagagg atctggcagc aaatgatatc ctggattaca     5401 aggatgacga cgataaggtt taaacggccg gccgcggtca tagctgtttc ctgaacagat     5461 cccgggtggc atccctgtga cccctcccca gtgcctctcc tggccctgga agttgccact     5521 ccagtgccca ccagccttgt cctaataaaa ttaagttgca tcattttgtc tgactaggtg     5581 tccttctata atattatggg gtggaggggg gtggtatgga gcaaggggca agttgggaag     5641 acaacctgta gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt ggcacaatct     5701 tggctcactg caatctccgc ctcctgggtt caagcgattc tcctgcctca gcctcccgag     5761 ttgttgggat tccaggcatg catgaccagg ctcagctaat ttttgttttt ttggtagagg     5821 cggggtttca ccatattggc caggctggtc tccaactcct aatctcaggt gatctaccca     5881 ccttggcctc ccaaattgct gggattacag gcgtgaacca ctgctccctt ccctgtcctt     5941 ctgattttaa aataactata ccagcaggag gacgtccaga cacagcatag gctacctggc     6001 catgcccaac cggtgggaca tttgagttgc ttgcttggca ctgtcctctc atgcgttggg     6061 tccactcagt agatgcctgt tgaattgggt acgcggccag cggcgagcgg tatcagctca     6121 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtc     6181 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac     6241 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg     6301 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac     6361 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat     6421 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag     6481 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac     6541 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt     6601 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt     6661 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc     6721 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga     6781 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac     6841 gacgcgtaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta     6901 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg     6961 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg     7021 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc     7081 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc     7141 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc     7201 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag     7261 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat     7321 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg     7381 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt     7441 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag     7501 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg     7561 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt     7621 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct     7681 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac     7741 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat     7801 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat     7861 ttatcagggt tattgtctca tgatgatatt attttatctt gtgcaatgta acatcagaga     7921 ttttgagaca cgggccagag ctgccaggaa acagctatga ccatgtaata cgactcacta     7981 taggggatat cagctggatg gcagttaac