Basic Vector Information
- Vector Name:
- BPK1520
- Antibiotic Resistance:
- Ampicillin
- Length:
- 2280 bp
- Type:
- Mammalian Expression, CRISPR
- Replication origin:
- ori
- Copy Number:
- High Copy
- Promoter:
- U6
- Cloning Method:
- Gibson Cloning
- 5' Primer:
- OS280 (5'-CAGGGTTATTGTCTCATGAGCGG-3')
BPK1520 vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
BPK1520 vector Sequence
LOCUS 40924_350 2280 bp DNA circular SYN 13-MAY-2021 DEFINITION Human expression plasmid for SpCas9 sgRNA (need to clone in spacer into BsmBI sites): U6-BsmBIcassette-Sp-sgRNA. ACCESSION . VERSION . KEYWORDS . 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) TITLE Direct Submission REFERENCE 3 (bases 1 to 2280) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nature. 2015 Jun 22. doi: 10.1038/nature14592." COMMENT SGRef: number: 2; type: "Journal Article" FEATURES Location/Qualifiers source 1..2280 /mol_type="other DNA" /organism="synthetic DNA construct" rep_origin complement(83..671) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(845..1702) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(1703..1807) /label=AmpR promoter promoter 1914..2154 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" misc_RNA 2184..2259 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system"
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