pAC94-pmax-dCas9VP160-2A-puro vector (V012298)

LOCUS       pAC94-pmax-dCas9        8282 bp DNA     circular SYN 01-JAN-1980
DEFINITION  CRISPR activation vector for expressing a puromycin resistance 
            marker plus catalytically inactive dCas9 fused to the VP160 
            transcriptional activation domain.
ACCESSION   .
VERSION     .
KEYWORDS    pAC94-pmax-dCas9VP160-2A-puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8282)
  AUTHORS   Cheng AW, Wang H, Yang H, Shi L, Katz Y, Theunissen TW, Rangarajan 
            S, Shivalila CS, Dadon DB, Jaenisch R.
  TITLE     Multiplexed activation of endogenous genes by CRISPR-on, an 
            RNA-guided transcriptional activator system.
  JOURNAL   Cell Res. 2013;23:1163-71.
  PUBMED    23979020
REFERENCE   2  (bases 1 to 8282)
  AUTHORS   Jaenisch Lab / Addgene #48226
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8282)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Res.";
            date: "2013"; volume: "23"; pages: "1163-71"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8282
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        139..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..719
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     intron          813..945
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     protein_bind    960..984
                     /label=attB1
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1005..1007
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             1008..1034
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="HA"
                     /translation="YPYDVPDYA"
     CDS             1038..1058
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T
                     antigen"
                     /note="SV40 NLS"
                     /translation="PKKKRKV"
     CDS             1068..5168
                     /label=dCas9
                     /note="catalytically dead mutant of the Cas9 endonuclease
                     from the Streptococcus pyogenes Type II CRISPR/Cas system"
     CDS             5172..5192
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T
                     antigen"
                     /note="SV40 NLS"
                     /translation="PKKKRKV"
     CDS             5226..5609
                     /label=VP160
                     /note="10 tandem repeats of the minimal activation domain
                     of herpes simplex virus VP16 (Cheng et al., 2013)"
     CDS             5634..5687
                     /label=T2A
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             5697..6293
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     protein_bind    complement(6312..6336)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             6389..6430
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
     polyA_signal    complement(6485..6566)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(6764..7351)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7435..8226)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"