Purpose(Empty Backbone) 2nd gen bicistronic lentiviral vector allows for simultaneous expression of a transgene and EGFP marker to facilitate tracking of transduced cells.
- Backbone size (bp) 11101
Vector typeMammalian Expression, Lentiviral, Cre/Lox
Growth in Bacteria
Growth instructionsUse Stbl3 or HB101 to reduce chance of recombination. Grow at 37C
Copy numberHigh Copy
/ Fusion Protein
- EMCV IRES-EGFP cassette (C terminal on backbone)
- Cloning method Restriction Enzyme
- 5′ cloning site MCS - see map (not destroyed)
- 3′ cloning site MCS - see map (not destroyed)
- 5′ sequencing primer See map (Common Sequencing Primers)
Terms and Licenses
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
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:pWPI was a gift from Didier Trono (Addgene plasmid # 12254)
Map generated by Addgene from full sequence supplied by depositor.
Map uploaded by the depositor.