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Minimal Promoter

OG251 - pSF-pA-MinProm-daGFP

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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.

Product Code: OG251

Quantity Provided: 5µg

Size (bp): 4717 bp

Bacterial Antibiotic Selection: Kanamycin

Origin and Compatibility: pUC high copy, derived from pBR322   

Copy Number: 500-700 copies per cell

Promoter:  Minimal promoter from the Herpes Simplex Thymidine Kinase Gene

Purpose: This plasmid contains a multiple cloning site that is upstream of a minimal promoter that is upstream of the chloramphenicol acetyl transferase (CAT) reporter gene. This plasmid can be used to insert specific DNA sequences that will endow the minimal promoter with specific biochemical properties such as tissue specificity or transcription factor specific activation.  

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.

daGFP: This vector encodes a green fluorescent protein called daGFP. This protein can be used in the same way as normal GFP using argon laser based or UV based excitation apparatus to allow the detection of fluorescence. The protein has a peak excitation of 510nm and a peak emission of 521nm. The gene encoding the protein is sold free of intellectual property restrictions.

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.
  • daGFP – The daGFP reporter gene has been modified to remove PciI and BseRI restriction sites.  

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.