p3s-Cas9HC vector (V012315)

MSDS

Basic Vector Information

Vector Name:: p3s-Cas9HC

Antibiotic Resistance:: Ampicillin

Length:: 7383 bp

Type:: CRISPR Plasmids

Replication origin:: ori

Source/Author:: Cho SW, Kim S, Kim JM, Kim JS.

Copy Number:: High copy number

p3s-Cas9HC 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 ℃.

p3s-Cas9HC vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       p3s-Cas9HC.        7383 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Mammalian cell vector for expressing S. pyogenes Cas9 with a 
            C-terminal HA epitope tag.
ACCESSION   .
VERSION     .
KEYWORDS    p3s-Cas9HC.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7383)
  AUTHORS   Cho SW, Kim S, Kim JM, Kim JS.
  TITLE     Targeted genome engineering in human cells with the Cas9 RNA-guided 
            endonuclease.
  JOURNAL   Nat. Biotechnol. 2013;31:230-2.
  PUBMED    23360966
REFERENCE   2  (bases 1 to 7383)
  AUTHORS   Kim Lab / Addgene #43945
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7383)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. 
            Biotechnol."; date: "2013"; volume: "31"; pages: "230-2"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7383
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        863..881
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             901..5004
                     /label=Cas9
                     /note="Cas9 (Csn1) endonuclease from the Streptococcus
                     pyogenes Type II CRISPR/Cas system"
     CDS             5020..5040
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T
                     antigen"
                     /note="SV40 NLS"
                     /translation="PKKKRKV"
     CDS             5041..5067
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
     promoter        complement(5101..5119)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    5145..5369
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(5628..6216)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6390..7247)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7248..7352)
                     /label=AmpR promoter

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