Basic Vector Information
BPK3079 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 ℃.
BPK3079 vector Sequence
LOCUS 40924_370 2225 bp DNA circular SYN 13-MAY-2021 DEFINITION Human expression plasmid for AsCpf1 guide RNA (need to clone in spacer into BsmBI sites). ACCESSION . VERSION . KEYWORDS . 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) TITLE Direct Submission REFERENCE 3 (bases 1 to 2225) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat Biotechnol. 2016 Jun 27. doi: 10.1038/nbt.3620." COMMENT SGRef: number: 2; type: "Journal Article" FEATURES Location/Qualifiers source 1..2225 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 82..322 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" rep_origin complement(476..1064) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(1238..2095) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(2096..2200) /label=AmpR promoter
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