EN.GFP vector (V009995)

Basic Vector Information

      • Vector Name:
      • EN.GFP
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 10655 bp
      • Type:
      • Cloning vector
      • Source/Author:
      • Chen SX, Osipovich AB, Ustione A, Potter LA, Hipkens S, Gangula R, Yuan W, Piston DW, Magnuson MA.

EN.GFP vector Vector Map

EN.GFP10655 bp5001000150020002500300035004000450050005500600065007000750080008500900095001000010500T3 promoterM13 revlac operatorlac promoterCAP binding siteoriAmpRAmpR promoterf1 oriM13 fwdT7 promoterLox66 siteRosa 26 promoter5'UTReGFPrabbit beta globin intron poly A signalFRT sitePGK promoterNeorabbit beta globin poly A signalFRT siteLox2272 site

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 ℃.

EN.GFP vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported               10655 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector EN.GFP, complete sequence.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10655)
  AUTHORS   Chen SX, Osipovich AB, Ustione A, Potter LA, Hipkens S, Gangula R, 
            Yuan W, Piston DW, Magnuson MA.
  TITLE     Quantification of factors influencing fluorescent protein expression
            using RMCE to generate an allelic series in the ROSA26 locus in mice
  JOURNAL   Dis Model Mech 4 (4), 537-547 (2011)
  PUBMED    21324933
REFERENCE   2  (bases 1 to 10655)
  AUTHORS   Chen SX, Osipovich AB, Magnuson MA.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-JUL-2010) Molecular Physiology and Biophysics, 
            Vanderbilt University, 9475 MRB IV, 2214 Garland Avenue, Nashville, 
            TN 37232, USA
REFERENCE   3  (bases 1 to 10655)
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10655
                     /organism="Cloning vector EN.GFP"
                     /mol_type="other DNA"
     promoter        complement(13..31)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(52..68)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
     protein_bind    76..92
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(100..130)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    145..166
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(454..1042)
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
     CDS             complement(1213..2073)
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
     promoter        complement(2074..2178)
                     /label=AmpR promoter
     rep_origin      complement(2253..2708)
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     2850..2866
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
     promoter        2873..2891
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2906..2939
                     /label=Lox66 site
                     /note="Lox66 site"
     protein_bind    complement(2906..2939)
                     /bound_moiety="Cre recombinase"
                     /note="Left element (LE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     regulatory      2957..7037
                     /note="Rosa 26 promoter"
     5'UTR           7043..7101
                     /label=translational enhancer
                     /note="translational enhancer"
     5'UTR           7050..7092
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     CDS             7103..7819
                     /note="green fluorescent protein"
     regulatory      7833..8718
                     /note="rabbit beta globin intron poly A signal"
     intron          7847..8419
                     /label=-beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     polyA_site      8616..8671
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
     misc_feature    8734..8767
                     /label=FRT site
                     /note="FRT site"
     protein_bind    8734..8767
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     regulatory      8815..9407
                     /note="PGK promoter"
     promoter        8831..9330
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     promoter        9342..9389
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9408..10211
     regulatory      10225..10526
                     /note="rabbit beta globin poly A signal"
     polyA_signal    10249..10473
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    10531..10564
                     /label=FRT site
                     /note="FRT site"
     protein_bind    10531..10564
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     misc_feature    complement(10617..10650)
                     /label=Lox2272 site
                     /note="Lox2272 site"
     protein_bind    complement(10617..10650)
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites."

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