pHAGE-CMV-dsRed-UBC-GFP-W
(Plasmid #24526)

• Purpose: (Empty Backbone)
• Depositing Lab: Darrell Kotton
• Publication: Wilson et al Am J Respir Cell Mol Biol. 2008 Aug . 39(2):133-41.

Backbone

• Vector backbone: pHAGE
• Backbone manufacturer: Richard Mulligan
• Vector type: Lentiviral

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Stbl3
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: None
• Tags / Fusion Proteins:
  • DsRed (N terminal on backbone)
  • GFP (C terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ sequencing primer: CMV-F, hUBCpro-F

Resource Information

• Supplemental Documents:
  • pHAGE-CMV-dsRED-UBC-GFP
• Addgene Notes:
  • Addgene diagnostic digest
  • Addgene Diagnostic Digest
• Articles Citing this Plasmid:
  • 10 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry

Depositor Comments

The pHAGE vector was modified for dual transgenesis as follows: cDNA encoding a variant of the red fluorescent protein adapted from Discosoma sp. (DsRed-Express; Clontech, Mountain View, CA) was amplified by PCR attaching NotI and BamH1 restriction sites to 5' and 3' ends, respectively. This amplicon was cloned into the pHAGE backbone in the first gene expression position by ligation to NotI/BamH1 cohesive ends. Next, enhanced green fluorescence protein (GFP; Clontech) cDNA was generated by PCR attaching NdeI and ClaI sites to the 5' and 3' ends, respectively, for ligation into the second gene position of pHAGE. Immediately upstream of the dsRed or GFP ATG start site, the indicated promoter fragment (cytomegalovirus [CMV], 584 bp; ubiquitin C [UBC], 397 bp]) was inserted by standard cloning techniques. See map for additional details.