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Multiple Cloning Site to Insert a Promoter

OG371 - pSF-PromMCS-SEAP

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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-PromMCS-SEAP

Product Code: OG371

Quantity Provided: 5µg

Size (bp): 5337

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 a multiple cloning site in the promoter position designed to allow you to insert your own promoter to drive the expression of the reporter gene. In this plasmid it is upstream of the human secreted alkaline phosphatase (SEAP) reporter gene. The promoter multiple cloning site extends from the SalI restriction site to the BstBI restriction site. Downstream sites have other functions in our plasmid system, for example, adding n-terminal peptide tags.  

Poly A signals: Unlike pSF-pA-PromMCS-SEAP, this plasmid does not contain a synthetic polyadenylation signal immediately upstream of the promoter multiple cloning site to reduce background transcription into the promoter MCS. This makes this plasmid useful when transferring any promoter into retroviral vector because the presence of a polyA can prevent full length virus genome production by causing premature transcription termination.  

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:

  • 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.
  • 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 region, 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.