| 出品公司: | ORIGENE |
|---|---|
| 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: | GE100001V |
| 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
载体基本信息
载体图谱质粒图谱和多克隆位点信息
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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