p3s-Cas9HC vector (V012315)

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 Map

p3s-Cas9HC7383 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300660069007200CMV enhancerCMV promoterT7 promoterCas9SV40 NLSHASP6 promoterbGH poly(A) signaloriAmpRAmpR promoter

p3s-Cas9HC vector Sequence

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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