| 质粒名称: | BPK1520 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 65777 |
| 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: | SpCas9 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: | BPK1520 was a gift from Keith Joung (Addgene plasmid # 65777 ; http://n2t.net/addgene:65777 ; RRID:Addgene_65777) |
| 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 |
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
载体序列
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
http://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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