Addgene - pHAGE-CMV-dsRed-UBC-GFP-W Plasmid Data

Plasmid 24526: pHAGE-CMV-dsRed-UBC-GFP-W

Gene/insert name:
None
Fusion protein or tag:
DsRed
Terminal:
N terminal on backbone
Fusion protein or tag:
GFP
Terminal:
C terminal on backbone
Vector backbone:
pHAGE
(Search Vector Database)
Backbone manufacturer:
Richard Mulligan
Vector type:
Lentiviral
5' sequencing primer:
CMV-F, hUBCpro-F List of Sequencing Primers
Bacterial resistance(s)
Ampicillin
Growth strain(s)
Stbl3
Growth temperature (℃):
37
High or low copy:
High Copy
Sequence: View sequences (3)
Map:
View map
Supplemental document:
pHAGE-CMV-dsRED-UBC-GFP (application/pdf)
Supplemental document:
Notes from Addgene (1)
Principal Investigator:
Darrell Kotton
Terms and Licenses:
MTA
Clontech Limited Use Label License

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.

Addgene has sequenced a portion of this plasmid for verification. Click here for the sequencing result.

Article: Sustained expression of alpha1-antitrypsin after transplantation of manipulated hematopoietic stem cells. Wilson et al (Am J Respir Cell Mol Biol. 2008 Aug . 39(2):133-41. PubMed)

Please acknowledge the principal investigator and cite this article if you use this plasmid in a publication. Also, please include the text "Addgene plasmid 24526" in your Materials and Methods section.