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SEAP

OG365 - pSF-CMVe-SEAP

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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-SEAP

Product Code: OG365

Quantity Provided: 5µg

Size (bp): 5825

Bacterial Antibiotic Selection: Kanamycin

Origin and Compatibility: pUC high copy, derived from pBR32

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 human secreted alkaline phosphatase (SEAP) reporter gene. 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-SEAP, 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.

Intellectual Property Status: According to our IP-friendly policy, this plasmid is sold free of reach-through rights on any derivatives you may create.

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.
  • SEAP Reporter Gene – The SEAP gene has been modified to remove SalI, Hind3, SphI, BamHI, NcoI, NdeI, Avr2, Sac2, NcoI, BspHI, StuI, EagI sites and two SacI sites, two NarI sites, two SmaI sites, as well as three BseRI sites and six BsgI sites to ensure compatibility with all of our products.

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.