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Constitutive

OG411 - Dual CMV Expression Plasmid - pSF-CMV-CMV-SbfI

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This plasmid is compatible with all of the other DNA sequences available on this site, which are all sold in the same backbone. This flexibility allows you to create any vector you require, with a variety of functions, simply by compiling the relevant sections from our product range.

Vector: pSF-CMV-CMV SbfI

Product Code: OG411

Quantity Provided: 5µg

Size (bp): 5131

Bacterial Antibiotic Selection: Kanamycin

Origin and Compatibility: pUC high copy, derived from pBR322   

Copy Number: 500-700 copies per cell

Promoter: Cytomegalovirus (CMV) immediate early promoter

Purpose: A versatile expression vector for the production of two proteins under the control of two separate CMV promoter expression cassettes in mammalian cells. This vector also contains a Kanamycin resistance cassette for growth and maintenance in E.coli.

Quality Validation: This vector has been demonstrated to express reporter genes to high levels under both of the CMV promoters. These reporter genes were inserted with the start codon and stop codon residing within the NcoI and XbaI site restriction site of the MCS that is downstream of the first (upstream) CMV promoter. Under the control of the second (downstream) CMV promoter the start codon and stop codon resided within the PciI and SpeI sites respectively. Expression was validated in 293 cells.  

Quality Sequencing Analysis: This vector has been fully sequenced using primers F1-F10. The details and sequences of these primers are available through our website. 

Genetic Modifications to standard parts: 
•    CMV promoters – An NcoI site has been ablated in both the CMV promoters. This change does not reduce expression. 
•    KanR cassette – This KanR resistance region is not a wild type sequence, it has been modified to remove all restriction sites that conflict with the SnapFast system and to retain a high level of antibiotic resistance. It has been validated in E.coli.

Restriction site notes: 
•    Bgl2, KpnI, SwaI and PmeI each cut the vector at two sites positioned to flank the promoter, start codon, origin, and KanR, respectively. 
•    BsgI and BseRI recognise non-palindromic DNA sequences and cleave upstream of their recognition sites, within the stop codon that is found inside the XbaI restriction site.