pEF-I-GFP GX
(Plasmid #45443)

• Purpose: (Empty Backbone)
• Depositing Lab: John Brigande
• Publication: Gubbels et al Nature. 2008 Sep 25. 455(7212):537-41.

Backbone

• Vector backbone: pEF-GFP (Addgene Plasmid #11154)
• Backbone manufacturer: Connie Cepko lab (Harvard Medical School, Boston, MA)
• Backbone size (bp): 7349
• Modifications to backbone: Converted to Gateway-compatible destination vector (gateway casette added). IRES inserted upstream of EGFP.
• Vector type: Mammalian Expression
• Promoter: human EF1α
• Selectable markers: EGFP
• Tag / Fusion Protein:
  • IRES EGFP (C terminal on backbone)

Growth in Bacteria

• Bacterial Resistance(s): Chloramphenicol and Ampicillin, 25 & 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): ccdB Survival
• Copy number: High Copy

Cloning Information

• Cloning method: Gateway Cloning
• 5′ sequencing primer: EF1a-F
• 3′ sequencing primer: pCDH-rev (GCATTCCTTTGGCGAGAG)

Resource Information

• A portion of this plasmid was derived from a plasmid made by: pEF-GFP backbone from Connie Cepko (Harvard Medical Schoo, Boston, MA). IRES-GFP fragment from plasmid pBMN-I-GFP from Gary Nolan (Stanford, CA). RfC.1 cassette from Invitrogen (Carlsbad, CA).
• Articles Citing this Plasmid:
  • 3 References

Terms and Licenses

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

Depositor Comments

Alternate plasmid name: pEF1α-RfC.1-IRES-GFP

To construct this vector, the depositing lab used EcoRI and BsrGI to replace the EGFP in pEF–GFP (Addgene plasmid 11154) with the IRES-GFP fragment from plasmid pBMN-I-GFP (Addgene plasmid 1736), creating intermediate vector pEF1α-IRES-GFP.

Plasmid pEF1α-IRES-GFP was then converted to a Gateway-enabled destination vector with the Gateway Vector Conversion System (Invitrogen) by digesting with EcoRI, blunting the ends, and inserting Reading Frame Cassette C (RfC.1) gateway cassette.