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pCas-Guide-AAVS1载体

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    • 技术资料
    • 保存条件

      -20℃低温保存

    • 保质期

      三年

    • 英文名

      pCas-Guide-AAVS1

    • 库存

      20

    • 供应商

      泽叶生物

    载体基本信息

    出品公司: ZYbscience
    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: --
    产品目录号: ZY100001V
    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 site
    AAVS1 target sequence: GTTAATGTGGCTCTGGTTCT
    CMV-driven Cas9 and U6-driven AAVS1 gRNA expression
    

    载体序列

    LOCUS       Exported File           8009 bp ds-DNA    circular SYN 23-七月-2016
    DEFINITION  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-AAVS1
    SOURCE      synthetic DNA construct
      ORGANISM  synthetic DNA construct
    REFERENCE   1  (bases 1 to 8009)
      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..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
    //
    
    

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