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.

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