| 质粒名称: | VVT1 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 65779 |
| 存储实验室: | Keith Joung |
|
载体骨架基本信息 |
|
| 载体名称: | MLM3636 |
| 载体出品公司: | Joung lab (Addgene Plasmid # 43860) |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达,CRISPR |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | SaCas9 gRNA backbone, without spacer sequence |
| 物种来源: | Synthetic |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | U6 |
| 标签/融合蛋白: | -- |
| 克隆方法: | Gibson克隆 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | OS280 (5'-CAGGGTTATTGTCTCATGAGCGG-3') |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | VVT1 was a gift from Keith Joung (Addgene plasmid # 65779 ; http://n2t.net/addgene:65779 ; RRID:Addgene_65779) |
| 参考文献部分: | Engineered CRISPR-Cas9 nucleases with altered PAM specificities. Kleinstiver BP, Prew MS, Tsai SQ, Topkar VV, Nguyen NT, Zheng Z, Gonzales AP, Li Z, Peterson RT, Yeh JJ, Aryee MJ, Joung JK. Nature. 2015 Jun 22. doi: 10.1038/nature14592. 10.1038/nature14592 PubMed 26098369 |
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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
载体序列
LOCUS VVT1 2324 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION The Joung Lab recommends using BPK2660 (Addgene plasmid 70709) instead of VVT1 as guide RNAs cloned into BPK2660 are more effective than this plasmid. Human expression plasmid for S. aureus Cas9 sgRNA. ACCESSION . VERSION . KEYWORDS VVT1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2324) AUTHORS Kleinstiver BP, Prew MS, Tsai SQ, Topkar VV, Nguyen NT, Zheng Z, Gonzales AP, Li Z, Peterson RT, Yeh JJ, Aryee MJ, Joung JK TITLE Engineered CRISPR-Cas9 nucleases with altered PAM specificities. JOURNAL Nature. 2015 Jun 22. doi: 10.1038/nature14592. PUBMED 26098369 REFERENCE 2 (bases 1 to 2324) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2324 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 82..322 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 82..102 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 253..272 /label=LKO.1 5' /note="Human U6 promoter, forward primer" rep_origin complement(575..1163) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(655..674) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1334..2194) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 1957..1976 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2195..2299) /gene="bla" /label=AmpR promoter ORIGIN 1 gacgtcgcta gctgtacaaa aaagcaggct ttaaaggaac caattcagtc gactggatcc 61 ggtaccaagg tcgggcagga agagggccta tttcccatga ttccttcata tttgcatata 121 cgatacaagg ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta 181 gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta 241 tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct 301 ttatatatct tgtggaaagg acgaaacacc cgagacgatt aatgcgtctc ggttttagta 361 ctctgtaatt ttaggtatga ggtagacgaa aattgtactt atacctaaaa ttacagaatc 421 tactaaaaca aggcaaaatg ccgtgtttat ctcgtcaact tgttggcgag atttttttaa 481 gcttgggccg ctcgaggtac ctctctacat atgacatgtg agcaaaaggc cagcaaaagg 541 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 601 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 661 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 721 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 781 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 841 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 901 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 961 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag 1021 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 1081 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta 1141 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc 1201 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca 1261 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa 1321 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat 1381 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct 1441 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt 1501 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat 1561 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta 1621 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg 1681 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt 1741 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg 1801 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg 1861 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc 1921 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa 1981 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac 2041 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt 2101 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg 2161 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa 2221 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata 2281 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acct //
