EF.GFP
(Plasmid #17616)

• Depositing Lab: Linzhao Cheng
• Publication: Cui et al Blood. 2002 Jan 15. 99(2):399-408.

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, 100 μg/mL
• 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
• Entrez Gene: EEF1A1 (a.k.a. CCS-3, CCS3, EE1A1, EEF-1, EEF1A, EF-Tu, EF1A, EF1A1, EF1alpha1, GRAF-1EF, LENG7, PTI1, eEF1A-1)
• Tag / Fusion Protein:
  • GFP (C terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ sequencing primer: EGFP-N

Resource Information

• A portion of this plasmid was derived from a plasmid made by: SIN LV pRLLhPGK.GFP Sin-18 (abbreviated as PGK.GFP) was kindly provided by Dr. Didier Trono.
• Articles Citing this Plasmid:
  • 5 References

Terms and Licenses

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

Depositor Comments

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.