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CMV Enhancer

OG363 - pSF-CMVe-FLuc

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Full Plasmid Details (PDF)

Sequence (txt)

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-CMVe-FLuc

Product Code: OG363

Quantity Provided: 5µg

Size (bp): 5912

Bacterial Antibiotic Selection: Kanamycin

Origin and Compatibility: pUC high copy, derived from pBR322 

Copy Number: 500-700 copies per cell

Promoter: Not applicable    

Purpose: This plasmid contains the major immediate early cytomegalovirus (CMV) enhancer region upstream of the Photinus pyralis (FLuc) luciferase reporter. The enhancer can be used to increase the strength of weaker promoters whilst often retaining any regulation that they already have, such as tissue specificity or transcription factor activation in response to stimulus, however, this is not always the case and will vary depending on the promoter being used.

Poly A signals: Unlike pSF-pA-CMVe-FLuc, this plasmid does not have a synthetic poly adenylation site between the CMV enhancer and the promoter multiple cloning site to reduce background transcription through to the promoter. This makes this plasmid useful when transferring the enhancer/promoter into retroviral vectors because the presence of a polyA can prevent full length virus genome production. The lack of upstream poly signal in this plasmid can lead to increased background activity, but this will vary depending on the promoter being used.  

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.

Restriction site notes:

  • Bgl2, SwaI and PmeI each cut the vector at two sites positioned to flank the promoter, 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.

Genetic Modifications to standard parts:

  • CMV enhancer – an NcoI site has been ablated in the CMV enhancer region. This change does not reduce its activity.
  • 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.
  • Photinus pyralis luciferase – The luciferase coding sequence has been modified to remove PspOMI, MreI, BbvCI sites as well as three BsgI sites and two BseRI sites.