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Plasmid 12254: pWPI
  • 2nd gen bicistronic lentiviral vector allows for simultaneous expression of a transgene and EGFP marker to facilitate tracking of transduced cells.

  • None

  • EMCV IRES-EGFP cassette

  • C terminal on backbone

  • pWPXL
    (Search Vector Database)

  • Mammalian Expression, Lentiviral, Cre/Lox

  • 11101

  • MCS - see map

  • No

  • MCS - see map

  • No

  • See map List of Sequencing Primers

  • Ampicillin

  • Stbl3

  • 37

  • Use Stbl3 or HB101 to reduce chance of recombination. Grow at 37C

  • High Copy

  • View sequences (5)
  • View map

  • Didier Trono

  • UBMTA
    Ancillary Agreement for Plasmids Containing FP Materials

Comments: 

This bicistronic vector allows for simultaneous expression of a transgene and EGFP marker to facilitate tracking of transduced cells. The EGFP marker cDNA has been inserted downstream of EMCV IRES.

A popular cloning strategy is to clone into the PmeI site upstream of the EMCV IRES. You can also clone into the PacI or SwaI site.

If you want to put another fluorescent protein after the IRES, note that you need to make sure that the open reading frame of the second gene is precisely fused to 11th ATG of IRES (i.e., 11th ATG of IRES is a start of your cDNA).

Please note that the full sequence for this plasmid is approximated and not fully verified. Please visit the Trono lab http://tronolab.epfl.ch for cloning strategies, protocols, publications, and more. See LentiWeb http://www.lentiweb.com for discussions on cloning strategies and protocols.

Updated sequence provided by Chang-Xin Shi, Mayo Clinic

Addgene has sequenced a portion of this plasmid for verification. Click here for the sequencing result.

Please acknowledge the principal investigator and cite this article if you use this plasmid in a publication. Also, please include the text "Addgene plasmid 12254" in your Materials and Methods section.

Price: US $65

Available to academic and non-profits only