pPK-CMV Fusion Vectors

• Modified hCMV immediate-early promoter provides high level constitutive expression in a wide variety of cells and cell types.
• Convenient choice of GFP or Luciferase reporter genes for control of expression experiments
• Two multiple cloning sites (MCS) for cloning gene of choice upstream or downstream of reporter gene (in fusion)
• Kanamycin/Neomycin resistance gene provide efficient vector selection in E. coli (with Kanamycin sulfate) or mammalian cells (with Neomycin sulfate)
• SV40 polyadenylation sign provides for stable mRNA by providing efficient transcription termination and polyadenylation
• pUC Origin of replication provides high copy number of vector in E. coli

The pPK-CMV Fusion Vectors have been designed for high-level expression of GFP or Luciferase fusion proteins and for creating GFP or Luciferase expressing stable cell lines. They contain an optimized CMV promoter and an intron sequence, which ensures extremely high constitutive expression in a wide variety of mammalian cell types. Due to two multiple cloning sites on either side of the reporter gene, N- or C-terminal fusion proteins can easily be generated. In addition, each vector contains a combined kanamycin/neomycin resistance gene, which allows forefficient selection in both E. coli (using kanamycin) and mammalian cells (using neomycin). An SV40 polyadenylation signal provides efficient transcription termination and polyadenylation - thus ensuring stable mRNA. A pUC origin of replication allows for high copy numbers in E. coli. The vectors can be used for either transient transfection or for generating stable cell lines expressing only the reporter gene or the fusion target-reporter gene construct.
Four versions of the vector are available for convenient and efficient gene cloning and expression of the target gene fused with a reporter gene:

pPK-CMV-F1 (C-GFP) has a GFP coding sequence with a stop codon inframe in the MCS. The gene of interest can therefore be expressed with GFP at the C-terminus.

pPK-CMV-F2 (N-GFP) has a GFP coding sequence without a stop codon inframe in the MCS. The gene of interest can therefore be expressed with GFP at the N-terminus.

pPK-CMV-F3 (C-Luc) has a Luciferase coding sequence with a stop codon inframe in the MCS. The gene of interest can therefore be expressed with Luciferase at the C-terminus.

pPK-CMV-F4 (N-Luc) has a Luciferase coding sequence without a stop codon inframe in the MCS. The gene of interest can therefore be expressed with Luciferase at the N-terminus.

See also: Cell-based Reporter Assays

See also: Cell Transfection Reagents