pIRES-EGFP-puro
(Plasmid #45567)

• Purpose: (Empty Backbone)
• Depositing Lab: Michael McVoy
• Publication: pIRES-EGFP-puro (unpublished)
• Citations: 24 Articles

Backbone

• Vector backbone: pEGFP-C1
• Backbone manufacturer: Clontech
• Backbone size (bp): 5729
• Modifications to backbone: NheI, XhoI, and SacI cloning sites and an encephalomyocarditis virus IRES inserted upstream of EGFP. Puromycin resistance protein (puromycin N-acetyl transferase) fused downstream of EGFP
• Vector type: Mammalian Expression
• Promoter: Phcmv (strong human cytomegalovirus immediate early promoter)
• Selectable markers: Puromycin ; EGFP
• Tag / Fusion Protein:
  • IRES-EGFP-Puro (C terminal on backbone)

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: pCDH-rev (GCATTCCTTTGGCGAGAG)

Resource Information

• Addgene Notes:
  • Addgene diagnostic digest

Terms and Licenses

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

Depositor Comments

Plasmid pIRES-EGFP-puro was constructed containing the Phcmv promoter followed by unique restriction sites (NheI, XhoI, and SacI) for insertion of genes. Downstream, an IRES from the encephalomyocarditis virus directs the translation of EGFP-puro from the same mRNA.

Addgene's sequencing results found differences compared to the depositing lab's sequence in the 3' region of the puro resistance marker. The resulting protein sequence matches NCBI WP_055528321.1 and should be functional.