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Photinus pyralis (Firefly) Luciferase Plasmids

OG388 - pSF-CMV-PGK-Fluc


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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-PGK-FLuc

Product Code: OG388

Quantity Provided: 5µg

Size (bp): 6394

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 and the mouse phosphoglycerate kinase promoter (PGK)

Purpose: This plasmid contains two promoters that terminate transcription at the same poly-adenylation signal, allowing the expression of two genes from one expression cassette, where the second gene is the Photinus pyralis luciferase (Firefly, FLuc) reporter gene. The second promoter (PGK) is approximately 10-  fold weaker than the upstream CMV promoter in most commonly used cell lines.  

Quality Validation: This vector has been demonstrated to express reporter genes to high levels under the CMV promoter. The start codon of these reporters was positioned within the NcoI site and the stop codon was positioned within the XbaI site. This plasmid also demonstrates intermediate levels of expression of luciferase from the PGK promoter. Expression has been validated in 293 cells.

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

Genetic Modifications to standard parts:

  • CMV promoter – an NcoI site has been ablated in the CMV promoter. 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.
  • PGK Promoter – The PGK promoter has been modified to remove three BsgI sites, two BseRI sites and one StuI site. It still contains a SpeI site because this is within a region that is important for promoter activity.
  • Photinus pyralis luciferase - The luciferase gene in this vector has been modified to remove three BsgI sites, two BseRI sites, one PspOMI site, one MreI site and one BbvCI site to ensure compatibility with all of our products.

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.