质粒名称: | BPK3079, pBPK3079-U6-AsCpf1crRNA |
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出品公司: | 爱迪基因 |
目录编号: | 78741 |
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: | AsCpf1 crRNA 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: | 限制性内切酶 |
5' 克隆位点: | BsmBI(未销毁) |
3' 克隆位点: | BsmBI(未销毁) |
5' 测序引物 | OS280 (5'-CAGGGTTATTGTCTCATGAGCGG-3') |
3' 测序引物 | -- |
文献引用方法 |
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材料和方法部分: | BPK3079 was a gift from Keith Joung (Addgene plasmid # 78741 ; http://n2t.net/addgene:78741 ; RRID:Addgene_78741) |
参考文献部分: | Genome-wide specificities of CRISPR-Cas Cpf1 nucleases in human cells. Kleinstiver BP, Tsai SQ, Prew MS, Nguyen NT, Welch MM, Lopez JM, McCaw ZR, Aryee MJ, Joung JK. Nat Biotechnol. 2016 Jun 27. doi: 10.1038/nbt.3620. 10.1038/nbt.3620 PubMed 27347757 |
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BPK3079载体 pBPK3079-U6-AsCpf1crRNA质粒基本信息
BPK3079质粒图谱 pBPK3079-U6-AsCpf1crRNA载体图谱和BPK3079载体序列 pBPK3079-U6-AsCpf1crRNA质粒序列多克隆位点信息
BPK3079质粒 pBPK3079-U6-AsCpf1crRNA载体简介
BPK3079质粒序列 pBPK3079-U6-AsCpf1crRNA载体序列
LOCUS BPK3079 2225 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION Human expression plasmid for AsCpf1 guide RNA (need to clone in spacer into BsmBI sites). ACCESSION . VERSION . KEYWORDS BPK3079 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2225) AUTHORS Kleinstiver BP, Tsai SQ, Prew MS, Nguyen NT, Welch MM, Lopez JM, McCaw ZR, Aryee MJ, Joung JK TITLE Genome-wide specificities of CRISPR-Cas Cpf1 nucleases in human cells. JOURNAL Nat Biotechnol. 2016 Jun 27. doi: 10.1038/nbt.3620. PUBMED 27347757 REFERENCE 2 (bases 1 to 2225) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2225 /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(476..1064) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(556..575) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1235..2095) /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 1858..1877 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2096..2200) /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 gtaatttcta ctcttgtaga tggagacgat 361 taatgcgtct ccttttttta agcttgggcc gctcgaggta cctctctaca tatgacatgt 421 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc 481 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa 541 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc 601 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg 661 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc 721 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc 781 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca 841 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact 901 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg 961 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt 1021 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct 1081 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga 1141 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa 1201 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac 1261 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga 1321 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc 1381 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca 1441 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta 1501 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg 1561 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc 1621 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg 1681 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt 1741 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt 1801 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata 1861 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc 1921 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac 1981 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa 2041 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct 2101 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat 2161 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc 2221 cacct //
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BPK3079质粒 pBPK3079-U6-AsCpf1crRNA载体应用举例
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