Expression, Reporter & Fusion Vectors
pPK-CMV Expression Vectors
Vectors for high-level gene expression
- High-level constitutive transgene expression
- Suitable for both stable (G418) and transient expression
- Expression of native and HA-tagged proteins
- Small size for increased transfection efficiency
- Convenient multiple cloning site containing the most commonly-used restriction sites
The pPK-CMV Expression Vectors contain a modified human cytomegalovirus (CMV) based promoter/enhancer sequence that has been optimized for high gene expression. Sequence motives that limit expression have been removed, while others that promote high expression evels have been retained. Consequently, these vectors yield significantly higher expression levels than other expression vectors such as pcDNA3.1 that are based on the standard CMV promoter.
In addition, the plasmid backbones are optimized to allow for very small vector sizes and higher plasmid yields when amplified in E. coli.
Three versions of the pPK-CMV Expression Vector are available:
pPK-CMV-E1 does not have an expression tag. It is intended for expressing recombinant proteins in the native forms.
pPK-CMV-E2 comes with an N-terminal HA (hemagglutinin) fusion tag.
pPK-CMV-E3 comes with a C-terminal HA-fusion tag. The HA fusion tags allow easy detection and purification of proteins.
All three Expression Vectors come with a positive control vector containing a CAT gene.
See also: Cell Transfection Reagents
pPK-CMV Reporter Vectors
For highest level transient gene/reporter gene expression in mammalian cells
- Suitable for in vitro and in vivo gene expression applications
- High plasmid yield in E. coli
- Expression ready with the following popular reporter genes: β-galactosidase, green fluorescent protein (GFP), luciferase, chloramphenicol acetyltransferase (CAT), secreted alkaline phosphatase (SEAP)
- Convenient multiple cloning site for optimized cloning and expression of target genes
The pPK-CMV Reporter Vectors have been designed for high level, transient expression of reporter or target genes. They contain an optimized CMV promoter followed by intron A from the CMV immediate-early gene (IE). This ensures extremely high transient expression in a wide variety of mammalian cells and tissues. In addition, the vectors contain an optimized plasmid backbone that has been extensively altered to achieve optimal plasmid production in E. coli.
The vectors are available without a reporter gene to clone and express a gene of interest (blank vector) as well as with the following reporter genes:
β-galactosidase (lacZ), green fluorescent protein (GFP), luciferase (Luc), chloramphenicol acetyltransferase (CAT), and secreted alkaline phosphatase (SEAP).
See also: Cell-based Reporter Assays
See also: Cell Transfection Reagents
pPK-CMV Fusion Vectors
High-level expression vectors for GFP or Luciferase fusion protein expression
- 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
Order Information
*kit content: 1 vial (25 µg) pPK-CMV Expression Vector and 1 vial (10 µg) pPK-CMV/CAT Positive Control Vector
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