- Backbone size (bp) 8220
Vector typeMammalian Expression, Lentiviral
Growth in Bacteria
- Cloning method Restriction Enzyme
- 5′ sequencing primer CMV-F (Common Sequencing Primers)
In a SIN LV containing the elongation factor 1alpha promoter, we included a second promoter from cytomegalovirus that drives expression of RFP as a reporter. Dual-promoter LVs can coexpress multiple transgenes efficiently in a single target cell.
Recombinant LVs were produced by transient transfection of the transducing vector into 293T cells with two packaging vectors: pMD.G, a plasmid expressing the VSV-G envelope gene, and pCMVDeltaR8.91, a plasmid expressing the HIV-1 gag/pol, tat, and rev genes.
There is an a->g mutation in this plasmid that abolishes the HpaI site. A 10bp insertion in Addgene sequence corresponding to position 6893, 21bp insertion in Addgene sequence at position 7405, and 11bp deletion in Addgene sequence at position 7544 do not alter function of the plasmid or RFP expression.
The uploaded author's map shows an example with GFP cloned downstream of the EF-1a promoter. The vector supplied by Addgene does not contain the GFP.
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:EF.CMV.RFP was a gift from Linzhao Cheng (Addgene plasmid # 17619)
For your References section:Lentiviral vectors with two independent internal promoters transfer high-level expression of multiple transgenes to human hematopoietic stem-progenitor cells. Yu X, Zhan X, D'Costa J, Tanavde VM, Ye Z, Peng T, Malehorn MT, Yang X, Civin CI, Cheng L. Mol Ther. 2003 Jun . 7(6):827-38. 10.1016/S1525-0016(03)00104-7 PubMed 12788657
Map generated by Addgene from full sequence supplied by depositor.
Map uploaded by the depositor.