EF.CMV.RFP
(Plasmid #17619)

• Purpose: (Empty Backbone)
• Depositing Lab: Linzhao Cheng
• Publication: Yu et al Mol Ther. 2003 Jun . 7(6):827-38.

Backbone

• Vector backbone: EF.CMV.RFP
• Backbone size (bp): 8220
• Vector type: Mammalian Expression, Lentiviral
• Selectable markers: RFP

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Stbl3
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: None

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ sequencing primer: CMV-F

Resource Information

• Addgene Notes:
  • Addgene Diagnostic Digest
• Articles Citing this Plasmid:
  • 14 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

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.

Information for Cloning Grade DNA (Catalog # 17619-DNA.cg) ( Back to top)

Purpose

Cloning grade DNA is suitable for use in PCR, cloning reactions, or transformation into E. coli. The purity and amount is not suitable for direct transfections.

Delivery

• Amount: 2 µg
• Guaranteed Concentration: 100 ng/µl +/- 5 ng/µl
• Pricing: $105 USD
• Storage: DNA can be stored at 4℃ (short term) or -20℃ (long term).

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Quality Control

Addgene has verified this plasmid using Next Generation Sequencing. Results are available here