| 质粒名称: | pSMVP-Cas9N |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 80934 |
| 存储实验室: | George Church |
|
载体骨架基本信息 |
|
| 载体名称: | pMAX |
| 载体出品公司: | Lonza |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 2900 bp |
| 完整质粒大小: | 5409 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达，CRISPR，合成生物学 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Cas9N,Split-Cas9 N-terminal half |
| 物种来源: | Synthetic |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pSMVP-Cas9N was a gift from George Church (Addgene plasmid # 80934 ; http://n2t.net/addgene:80934 ; RRID:Addgene_80934) |
| 参考文献部分: | A multifunctional AAV-CRISPR-Cas9 and its host response. Chew WL, Tabebordbar M, Cheng JK, Mali P, Wu EY, Ng AH, Zhu K, Wagers AJ, Church GM. Nat Methods. 2016 Sep 5. doi: 10.1038/nmeth.3993. 10.1038/nmeth.3993 PubMed 27595405 |
买家导航
pSMVPCas9N载体基本信息
pSMVPCas9N载体图谱质粒图谱和多克隆位点信息
pSMVPCas9N载体简介
pSMVPCas9N载体序列
LOCUS pSMVP-Cas9N 5409 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Plasmid that expresses Cas9N in mammalian cells..
ACCESSION .
VERSION .
KEYWORDS pSMVP-Cas9N
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5409)
AUTHORS Chew WL, Tabebordbar M, Cheng JK, Mali P, Wu EY, Ng AH, Zhu K,
Wagers AJ, Church GM
TITLE A multifunctional AAV-CRISPR-Cas9 and its host response.
JOURNAL Nat Methods. 2016 Sep 5. doi: 10.1038/nmeth.3993.
PUBMED 27595405
REFERENCE 2 (bases 1 to 5409)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5409
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(39..59)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
enhancer 289..668
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 669..872
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 823..843
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
intron 964..1096
/label=chimeric intron
/note="chimera between introns from human beta-globin and
immunoglobulin heavy chain genes"
regulatory 1126..1135
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1138..1158
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 1183..2898
/codon_start=1
/product="N-terminal portion of Streptococcus pyogenes Cas9
(Zetsche et al., 2015)"
/label=Cas9(N)
/translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE"
primer_bind complement(3638..3657)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
polyA_signal 3667..3748
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind 3692..3711
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
rep_origin complement(3946..4534)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4026..4045)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4615..5409)
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(4726..4745)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 5336..5355
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
ORIGIN
1 aatattattg aagcatttat cagggttcgt ctcgctgtgg aatgtgtgtc agttagggtg
61 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
121 agcaaccagt cccggtctcc tcccatgcat gtcaatattg gccattagcc atattattca
181 ttggttatat agcataaatc aatattggct attggccatt gcatacgttg tatctatatc
241 ataatatgta catttatatt ggctcatgtc caatatgacc gccatgttgg cattgattat
301 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
361 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
421 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
481 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
541 tgccaagtcc gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
601 agtacatgac cttacgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
661 ttaccatggt gatgcggttt tggcagtaca ccaatgggcg tggatagcgg tttgactcac
721 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
781 aacgggactt tccaaaatgt cgtaataacc ccgccccgtt gacgcaaatg ggcggtaggc
841 gtgtacggtg ggaggtctat ataagcagag gtcgtttagt gaaccgtcag atcactagta
901 gctttattgc ggtagtttat cacagttaaa ttgctaacgc agtcagtgct cgactgatca
961 caggtaagta tcaaggttac aagacaggtt taaggaggcc aatagaaact gggcttgtcg
1021 agacagagaa gattcttgcg tttctgatag gcacctattg gtcttactga catccacttt
1081 gcctttctct ccacaggggt accgaagccg ctagcgctac cggtcgccac catggcccca
1141 aagaagaagc ggaaggtcgg tatccacgga gtcccagcag ccgacaagaa gtactccatt
1201 gggctcgata tcggcacaaa cagcgtcggc tgggccgtca ttacggacga gtacaaggtg
1261 ccgagcaaaa aattcaaagt tctgggcaat accgatcgcc acagcataaa gaagaacctc
1321 attggcgccc tcctgttcga ctccggggaa acggccgaag ccacgcggct caaaagaaca
1381 gcacggcgca gatatacccg cagaaagaat cggatctgct acctgcagga gatctttagt
1441 aatgagatgg ctaaggtgga tgactctttc ttccataggc tggaggagtc ctttttggtg
1501 gaggaggata aaaagcacga gcgccaccca atctttggca atatcgtgga cgaggtggcg
1561 taccatgaaa agtacccaac catatatcat ctgaggaaga agcttgtaga cagtactgat
1621 aaggctgact tgcggttgat ctatctcgcg ctggcgcata tgatcaaatt tcggggacac
1681 ttcctcatcg agggggacct gaacccagac aacagcgatg tcgacaaact ctttatccaa
1741 ctggttcaga cttacaatca gcttttcgaa gagaacccga tcaacgcatc cggagttgac
1801 gccaaagcaa tcctgagcgc taggctgtcc aaatcccggc ggctcgaaaa cctcatcgca
1861 cagctccctg gggagaagaa gaacggcctg tttggtaatc ttatcgccct gtcactcggg
1921 ctgaccccca actttaaatc taacttcgac ctggccgaag atgccaagct tcaactgagc
1981 aaagacacct acgatgatga tctcgacaat ctgctggccc agatcggcga ccagtacgca
2041 gacctttttt tggcggcaaa gaacctgtca gacgccattc tgctgagtga tattctgcga
2101 gtgaacacgg agatcaccaa agctccgctg agcgctagta tgatcaagcg ctatgatgag
2161 caccaccaag acttgacttt gctgaaggcc cttgtcagac agcaactgcc tgagaagtac
2221 aaggaaattt tcttcgatca gtctaaaaat ggctacgccg gatacattga cggcggagca
2281 agccaggagg aattttacaa atttattaag cccatcttgg aaaaaatgga cggcaccgag
2341 gagctgctgg taaagcttaa cagagaagat ctgttgcgca aacagcgcac tttcgacaat
2401 ggaagcatcc cccaccagat tcacctgggc gaactgcacg ctatcctcag gcggcaagag
2461 gatttctacc cctttttgaa agataacagg gaaaagattg agaaaatcct cacatttcgg
2521 ataccctact atgtaggccc cctcgcccgg ggaaattcca gattcgcgtg gatgactcgc
2581 aaatcagaag agaccatcac tccctggaac ttcgaggaag tcgtggataa gggggcctct
2641 gcccagtcct tcatcgaaag gatgactaac tttgataaaa atctgcctaa cgaaaaggtg
2701 cttcctaaac actctctgct gtacgagtac ttcacagttt ataacgagct caccaaggtc
2761 aaatacgtca cagaagggat gagaaagcca gcattcctgt ctggagagca gaagaaagct
2821 atcgtggacc tcctcttcaa gacgaaccgg aaagttaccg tgaaacagct caaagaagac
2881 tatttcaaaa agattgaatg tttcgactct gttgaaatca gcggagtgga ggatcgcttc
2941 aacgcatccc tgggaacgta tcacgatctc ctgaaaatca ttaaagacaa ggacttcctg
3001 gacaatgagg agaacgagga cattcttgag gacattgtcc tcacccttac gttgtttgaa
3061 gatagggaga tgattgaaga acgcttgaaa acttacgctc atctcttcga cgacaaagtc
3121 atgaaacagc tcaagaggcg ccgatataca ggatgggggc ggctgtcaag aaaactgatc
3181 aatgggatcc gagacaagca gagtggaaag acaatcctgg attttcttaa gtccgatgga
3241 tttgccaacc ggaacttcat gcagttgatc catgatgact ctctcacctt taaggaggac
3301 atccagaaag cacaagtttg tctggctggc gatactctca ttaccctggc cgatggacga
3361 cgagtgccta ttagagaact ggtgtcacag cagaattttt ccgtgtgggc tctgaatcct
3421 cagacttacc gcctggagag ggctagagtg agtagagctt tctgtaccgg catcaaacct
3481 gtgtaccgcc tcaccactag actggggaga tccattaggg ccactgccaa ccaccgattt
3541 ctcacacctc agggctggaa acgagtcgat gaactccagc ctggagatta cctggctctg
3601 cctaggagaa tccctactgc ctcctgatct cgagctcgat gagtttggac aaaccacaac
3661 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
3721 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
3781 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtac
3841 ttaagagggg gagaccaaag ggcgagacgt taaggcctca cgtgacatgt gagcaaaagg
3901 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
3961 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
4021 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
4081 cctgccgctt acgggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
4141 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
4201 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
4261 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
4321 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
4381 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
4441 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
4501 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
4561 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatgc cgtctcagaa
4621 gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta
4681 aagcacgagg aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc
4741 caacgctatg tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga
4801 aaagcggcca ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag
4861 atcctcgccg tcgggcatgc tcgccttgag cctggcgaac agttcggctg gcgcgagccc
4921 ctgatgctct tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc
4981 tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg
5041 cagccgccgc attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga
5101 caggagatcc tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac
5161 aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc
5221 ctcgtcttgc agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg
5281 cccctgcgct gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca
5341 gtcatagccg aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg
5401 ttcaatcat
//
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pSMVPCas9N载体质粒应用举例
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