質粒名稱: | BPK1520 |
---|---|
出品公司: | 愛迪基因 |
目錄編號: | 65777 |
存儲實驗室: | Keith Joung |
載體骨架基本信息 |
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載體名稱: | MLM3636 |
載體出品公司: | Joung lab (Addgene Plasmid # 43860) |
載體抗性: | 氨芐青霉素 |
空載體大小: | -- |
完整質粒大小: | -- |
載體修飾: | -- |
載體類型: | 哺乳細胞表達,CRISPR |
篩選標記: | -- |
高拷貝/低拷貝: | 高拷貝 |
生長條件: | 37 ℃ |
插入基因信息 |
|
插入基因名稱: | SpCas9 gRNA backbone, without spacer sequence |
物種來源: | Synthetic |
插入大小: | -- |
基因突變: | -- |
GenBank編號 | -- |
啟動子 | U6 |
標簽/融合蛋白: | -- |
克隆方法: | Gibson克隆 |
5' 克隆位點: | -- |
3' 克隆位點: | -- |
5' 測序引物 | OS280 (5'-CAGGGTTATTGTCTCATGAGCGG-3') |
3' 測序引物 | -- |
文獻引用方法 |
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材料和方法部分: | BPK1520 was a gift from Keith Joung (Addgene plasmid # 65777 ; //n2t.net/addgene:65777 ; RRID:Addgene_65777) |
參考文獻部分: | 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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BPK1520載體質粒基本信息
BPK1520質粒圖譜載體圖譜和BPK1520載體序列質粒序列多克隆位點信息
BPK1520質粒載體簡介
BPK1520質粒序列載體序列
LOCUS BPK1520 2280 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Human expression plasmid for SpCas9 sgRNA (need to clone in spacer into BsmBI sites): U6-BsmBIcassette-Sp-sgRNA. ACCESSION . VERSION . KEYWORDS BPK1520 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2280) 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 2280) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 //www.snapgene.com FEATURES Location/Qualifiers source 1..2280 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin complement(83..671) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(163..182) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(842..1702) /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 1465..1484 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1703..1807) /gene="bla" /label=AmpR promoter promoter 1914..2154 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 1914..1934 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 2085..2104 /label=LKO.1 5' /note="Human U6 promoter, forward primer" misc_RNA 2184..2259 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" ORIGIN 1 cgaggtacct ctctacatat gacatgtgag caaaaggcca gcaaaaggcc aggaaccgta 61 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 121 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 181 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 241 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 301 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 361 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 421 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 481 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct 541 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 601 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa 661 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa 721 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt 781 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca 841 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca 901 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc 961 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa 1021 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc 1081 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca 1141 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat 1201 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag 1261 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac 1321 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt 1381 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt 1441 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc 1501 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat 1561 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca 1621 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga 1681 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg 1741 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg 1801 ttccgcgcac atttccccga aaagtgccac ctgacgtcgc tagctgtaca aaaaagcagg 1861 ctttaaagga accaattcag tcgactggat ccggtaccaa ggtcgggcag gaagagggcc 1921 tatttcccat gattccttca tatttgcata tacgatacaa ggctgttaga gagataatta 1981 gaattaattt gactgtaaac acaaagatat tagtacaaaa tacgtgacgt agaaagtaat 2041 aatttcttgg gtagtttgca gttttaaaat tatgttttaa aatggactat catatgctta 2101 ccgtaacttg aaagtatttc gatttcttgg ctttatatat cttgtggaaa ggacgaaaca 2161 ccggagacga ttaatgcgtc tccgttttag agctagaaat agcaagttaa aataaggcta 2221 gtccgttatc aacttgaaaa agtggcaccg agtcggtgct ttttttaagc ttgggccgct //
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