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BPK1520 vector (V006615)

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

BPK15202280 bp60012001800oriAmpRAmpR promoterU6 promotergRNA scaffold

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

Copy Sequence

Download GeneBank File(.gb)

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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