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EF.GFP
(Plasmid #17616)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 17616 Plasmid sent as bacteria in agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    SIN LV pRLLhPGK.GFP Sin-18 (modified)
  • Backbone manufacturer
    Dr. Didier Trono
  • Vector type
    Mammalian Expression, Lentiviral

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    Stbl3
  • Copy number
    Unknown

Gene/Insert

  • Gene/Insert name
    EF-1a promoter
  • Alt name
    EF-1 alpha promoter
  • Species
    H. sapiens (human)
  • Insert Size (bp)
    1200
  • GenBank ID
    J04616
  • Tag / Fusion Protein
    • GFP (C terminal on backbone)

Cloning Information

Resource Information

Depositor Comments

Addgene sequencing results have found a 10 bp insert between nt 6201 and 6202 of the depositor's full plasmid sequence and a C6203G mutation. These differences are not believed to affect the function of the plasmid.

SIN lentiviral vector that expresses GFP controlled by the promoter of the housekeeping gene EF-1alpha.

We constructed EF.GFP by replacing the human PGK promoter (EcoRV to BamHI sites in the PGK.GFP vector) with human EF-1alpha promoter from plasmid pEF1/Myc-His-A (Invitrogen, Carlsbad, CA). The EF-1alpha promoter (nucleotide 373 to 1560 as numbered in GenBank accession J04616) contains the first exon, first intron, and part of the second exon before the coding sequence (starting at nucleotide 1582).

Recombinant LVs were generated using the 3-plasmid system by cotransfection of 293T cells through calcium phosphate precipitation. The ratio of a transducing vector, pMD.G, and pCMVDeltaR8.91 was fixed at 1.5:0.5:2 µg for 10^6 293T cells.

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.GFP was a gift from Linzhao Cheng (Addgene plasmid # 17616)
  • For your References section:

    Targeting transgene expression to antigen-presenting cells derived from lentivirus-transduced engrafting human hematopoietic stem/progenitor cells. Cui Y, Golob J, Kelleher E, Ye Z, Pardoll D, Cheng L. Blood. 2002 Jan 15. 99(2):399-408. 10.1182/blood.V99.2.399 PubMed 11781219