3xFLAG-dCas9/pMXs-puro vector (V012351)

LOCUS       3xFLAG-dCas9_pMX       10003 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Retroviral vector with a puromycin resistance marker, encoding 
            FLAG(R)-tagged dCas9 for engineered ChIP (enChIP) purification of 
            specific genomic regions.
ACCESSION   .
VERSION     .
KEYWORDS    3xFLAG-dCas9 pMXs-puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10003)
  AUTHORS   Fujita T, Fujii H.
  TITLE     Identification of proteins associated with an IFN-gamma-responsive 
            promoter by a retroviral expression system for enChIP using CRISPR.
  JOURNAL   PLoS ONE 2014;9:e103084.
  PUBMED    25051498
REFERENCE   2  (bases 1 to 10003)
  AUTHORS   Fujii Lab / Addgene #51240
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 10003)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE"; 
            date: "2014"; volume: "9"; pages: "e103084"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..10003
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             22..615
                     /note="long terminal repeat from Moloney murine leukemia
                     virus"
     misc_feature    678..1035
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus
                     (MMLV)"
     CDS             1094..1510
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag
                     gene lacking the start codon"
     misc_feature    1520..1894
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     CDS             1948..1950
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             1951..2016
                     /label=3xFLAG
                     /note="three tandem FLAG(R) epitope tags, followed by an 
                     enterokinase cleavage site"
     CDS             2020..6123
                     /label=dCas9
                     /note="catalytically dead mutant of the Cas9 endonuclease
                     from the Streptococcus pyogenes Type II CRISPR/Cas system"
     CDS             6136..6156
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T
                     antigen"
                     /note="SV40 NLS"
                     /translation="PKKKRKV"
     promoter        6343..6672
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             6682..7278
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     LTR             7386..7977
                     /label=LTR
                     /note="long terminal repeat from Moloney murine leukemia
                     virus"
     rep_origin      complement(8278..8866)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(9033..9890)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(9891..9995)
                     /label=AmpR promoter