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EF.CMV.RFP
(Plasmid #17619)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 17619 Standard format: Plasmid sent in bacteria as agar stab 1 $85
Cloning Grade DNA 17619-DNA.cg 2 µg of cloning grade DNA in Tris buffer 1 $105

This material is available to academics and nonprofits only.

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

Resource Information

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.

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
  • Industry Terms
    • Not Available to Industry

Quality Control

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

How to cite this plasmid ( Back to top)

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 ; http://n2t.net/addgene:17619 ; RRID:Addgene_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