pGreenFire 2.0-CMV positive control plasmid (pGF2-CMV-rFluc-T2A-GFP-mPGK-Puro)

No need to make a positive control for your pGreenFire 2.0 projects—SBI’s already built one for you. With the pGreenFire 2.0 CMV Positive Control Lentivector (pGF2-CMV-rFluc-T2A-GFP-mPGK-Puro), the GreenFire 2.0 cassette is driven by the CMV promoter for strong expression in common cell lines such as HeLa, HEK293, and HT1080. As with all of our pGreenFire 2.0 lentivectors, the GreenFire cassette now consists of red firefly luciferase (rFLuc), a T2A co-expression element, and GFP. The switch to rFLuc opens up the possibility of performing a dual-spectral luciferase assay and also delivers greater sensitivity for in vivo applications than conventional luciferase.

The pGreenFire 2.0 CMV Positive Control Lentivector uses the same smart design as our pGreenFire 2.0 reporters, which adds in a constitutive selection cassette for stable cell line generation while minimizing interference with the upstream GreenFire cassette. By using a weak/moderate mPGK promoter to drive the antibiotic selection marker (puromycin resistance) and carefully arranging the reporter genes, the selection marker is reliably expressed along with rFLuc and GFP.

See our pGreenFire 2.0 transcriptional response element reporters in action

 

The pGreenFire 2.0 Positive Control Lentivector delivers robust expression in MDA-MB-213 cells. Relative luciferase activity (A) and GFP activity (B) both correlate with the amount of virus used. (C) Like all pGreenFire 2.0 lentivectors, the pGreenFire 2.0 Positive Control Lentivector contains an mPGK-Puro cassette to streamline creation of stable reporters integrated into the cell lines of your choice.