pDRIVE5-SEAP is a family of plasmids with an expanding collection of native and composite promoters.
Promoters are valuable tools to study the expression of a given gene both in vitro and in vivo. Now with pDRIVE, you can choose to express your gene of interest at high or low levels, ubiquitously or specifically, and in a constitutive or inducible manner.
pDRIVE5-SEAP plasmids feature an engineered secreted embryonic alkaline phosphatase (SEAP) reporter gene. SEAP expression levels can be assayed with chromogenic or luminescent methods.
These plasmids are selectable with Zeocin™ in E. coli.
Native or Composite Promoters
• Native promoters, also called minimal promoters, consist of a single fragment from the 5’ region of a given gene. Each of them comprises a core promoter and its natural 5’UTR. In some cases, the 5’UTR contains an intron.
• Composite promoters combine promoter elements of different origins (e.g. SV40 enhancer/AFP promoter) or were generated by assembling a distal enhancer with a minimal promoter of the same origin (e.g. CEA enhancer/promoter).
Various Expression Patterns
• Ubiquitous Promoters, strongly active in a wide range of cells, tissues and cell cycles.
• Tissue Specific Promoters, active in a specific type of cells or tissues.
• Tumor Specific Promoters, active specifically in tumor cells.
• SEAP gene encodes an engineered secreted embryonic alkaline phosphatase. The levels of SEAP in the culture medium of transfected cells expressing the reporter gene can be assayed with chromogenic or luminescent methods.
• SV40 pAn: The Simian Virus 40 late polyadenylation signal enables efficient cleavage and polyadenylation reactions resulting in high levels of steady-state mRNA.
• pMB1 Ori is a minimal E. coli origin of replication with the same activity as the longer Ori.
• EM2K is a bacterial promoter that enables the constitutive expression of the antibiotic resistance gene in E. coli.
• Sh ble gene confers zeocin resistance therefore allowing the selection of transformed E. coli carrying a pDRIVE5s plasmid. Note: Stable transfection of clones cannot be performed due to the absence of an eukaryotic promoter upstream of the Sh ble gene.
Easy Subcloning of the Promoters
In every pDRIVE5-SEAP, several unique restriction sites flank the promoter (a multiple cloning site at the 5’ end and Nco I or Bsp HI at the 3’ end) in order to excise the promoter easily. These restriction sites are compatible with many other enzymes, thus facilitating cloning. Other restriction sites may be introduced by performing PCR using primers containing the restriction sites of interest.
- 20 µg of lyophilized DNA.
- 4 pouches of E. coli Fast-Media® Zeo (2 TB and 2 Agar)