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OG373 - pSF-PromMCS-CAT

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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-PromMCS-CAT

Product Code: OG373

Quantity Provided: 5µg

Size (bp): 4431

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 chloramphenicol acetyl transferase (CAT) 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-CAT, 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.
  • CAT Reporter Gene – The CAT gene has been modified to remove an NcoI site and an EcoRI site to ensure compatibility with all of our products. These changes did not alter the amino acid sequence of the CAT protein.

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.