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OG104 - pSF-CMV-CAT

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Vector: pSF-CMV-CAT

Product Code: OG104

Quantity Provided:
 5µg

Size (bp):
 4870

Bacterial Antibiotic Selection:
 Ampicillin

Origin and Compatibility: 
pUC high copy, derived from pBR322   

Copy Number: 
500-700 copies per cell

Promoter: 
Cytomegalovirus (CMV) immediate early promoter. 

Purpose:
 The expression of the chloramphenicol acetyl transferase(CAT) reporter gene under the control of the CMV promoter. The CAT gene is within the primary multiple cloning site.

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 Validation: This vector has been demonstrated to express the CAT reporter gene to high levels under the CMV promoter. The start codon of these reporters was positioned within the NcoI site and the stop codon was positioned within the XbaI restriction site of pSF-CMV, although XbaI site was ablated by the insertion of reporter gene. Reporter gene expression was validated in the A549 lung carcinoma cell line. This is strongest CAT expression vector we currently sell.  

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:

  • CMV promoter – an NcoI site has been ablated in the CMV promoter. This change does not reduce expression.
  • AmpR cassette – This AmpR resistance region is not a wild type sequence, it has been modified to remove all restriction sites that conflict with the SnapFastTM 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, KpnI, SwaI and PmeI each cut the vector at two sites positioned to flank the promoter, start codon, origin, and AmpR, respectively.
  • BsgI and BseRI recognise non-palindromic DNA sequences and cleave upstream of their recognition sites, within the stop codon of the CAT gene. This allows the fusion of any of our C-terminal tags to the CAT gene.