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Minimal Promoter

OG362 - pSF-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.

Vector: pSF-MinProm-daGFP

Product Code: OG362

Quantity Provided: 5µg

Size (bp): 4561

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 daGFP fluorescent 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: Unlike pSF-pA-MinProm-daGFP, this plasmid does not have a synthetic poly adenylation site immediately upstream of the minimal promoter to reduce background transcription. This is useful when transferring the promoter into retroviral vectors because the presence of a polyA can prevent full length virus genome production.

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.  

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.
  • 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.