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SEAP

OG238 - pSF-pA-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: OG238

Quantity Provided: 5µg

Size (bp): 5395 bp

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: The promoter multiple cloning site region in this plasmid contains a synthetic poly-adenylation site. This reduces the level of background transcription from the plasmid. However, if you intend to insert this region into a lentivirus or retrovirus, the polyA signal will reduce the virus titre because of premature genome termination. We also have a variant of this plasmid without the polyA signal that can be used for this purpose if it is required.  

Transcription Termination: This plasmid contains three alternative transcription terminators for mammalian, bacterial and bacteriophage (T7) expression. This means that only the promoter needs to be changed to alter the expression system you are using. We sell multiple promoters that can be used in each of these systems. The presence of each terminator does not reduce expression in the alternative systems.

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

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:

  • 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, 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.