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OG105 - CMV SEAP Plasmid - pSF-CMV-SEAP

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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.

Product Code: OG105

Size (bp): 5849

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 human secreted alkaline phosphatase (SEAP) reporter gene under the control of the CMV promoter. The SEAP gene is within the primary multiple cloning site.

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.

C-Terminal Fusions: This plasmid contains the SEAP gene in the main multiple cloning site. Any of our C-terminal coding tags and sequences can be fused to the C-terminus of the SEAP gene in a single cloning step. This can be performed by cutting the vector with BsgI, or BseRI, and any other downstream restriction site (such as NheI). Cleaving any of our C-terminal tag plasmids (with the suffix 1 at the end of the name) with the same restriction sites will create compatible ends. When ligated together the stop codon of the SEAP gene will be removed, and the C-terminal tag will be in frame with the SEAP coding sequence.

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:

  • 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.
  • SEAP reporter gene – The SEAP gene has been modified to remove SalI, Hind3, SphI, BamHI, NcoI, NdeI, Avr2, Sac2, NcoI, BspHI, StuI, EagI sites and two SacI sites, two NarI sites, two SmaI sites, as well as three BseRI sites and six BsgI sites to ensure compatibility with all of our products. 

Restriction site notes:

  • Bgl2, KpnI, SwaI and PmeI each cut the plasmid 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 that is found inside the XbaI restriction site.