pIRES-EGFP-puro
(Plasmid #45567)

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• Publication: pIRES-EGFP-puro (unpublished)

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
• Articles Citing this Plasmid:
  • 22 References

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.

Information for Cloning Grade DNA (Catalog # 45567-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:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Quality Control

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