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pUC Origin Plasmids

OG207 - pSF-RecA-BetaGal


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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-RecA-BetaGal

Product Code: OG207

Quantity Provided: 5µg

Size (bp): 6969 bp

Bacterial Antibiotic Selection: Kanamycin

Origin and Compatibility: pUC high copy, derived from pBR322

Copy Number: 500-700 copies per cell

Promoter: Constitutive RecA bacterial promoter with the LexA repressor binding site deleted.

Purpose: The expression of the beta galactosidase (beta gal) reporter gene under the control of the RecA constitutive bacterial promoter. This plasmid can be used for monitoring reporter gene activity in bacterial cells or colonies using X-Gal which will produce a blue colouration. The reporter gene has been inserted thin the primary multiple cloning site. X-Gal is not membrane permeable and so cells must be lysed prior to detect, when screening for blue colonies on an agar plate sufficient cells will naturally lyse on the plate to allow blue colony detection.

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 Validation: This vector has been demonstrated to express the beta gal reporter gene to high levels under the RecA promoter. The start codon of the beta gal reporters was positioned within the NcoI site and the stop codon was positioned within the XbaI site. Expression was validated in the Escherichia coli.  

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:

  • RecA promoter – The LexA binding site has been ablated in this promoter to provide constitutive expression.  
  • 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