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OG159 - pSF-CMV-Ub-CAT-Ascl


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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: OG159

Size (bp): 6207

Bacterial Antibiotic Selection: Kanamycin

Origin and Compatibility: pUC high copy, derived from pBR322   

Copy Number: 500-700 copies per cell

Promoter: Cytomegalovirus (CMV) immediate early promoter and the Human Ubiquitin C promoter (Ub).

Purpose: This vector contains a Chloramphenicol Acetyl Transferase (CAT) expression cassette under the control of the Ub promoter that is designed to be used as an add-on to any of our vectors. The Ub expression cassette is flanked by AscI sites, but is not designed to be split up. If you would like the CAT gene alone, or the Ub promoter alone, please see the reporter gene section, or the promoter section, on our website. The Ub promoter is a relatively strong promoter. Its strength is approximately 10-fold lower than the CMV promoter, but 10-fold stronger than the SV40 promoter.  We have a weaker Rous Sarcoma virus promoter driven vector in the same format if you require an expression vector with lower levels of reporter gene activity.      

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.

Inserting a Gene: Each restriction site in this vector has a purpose and allows the insertion of specific pre-designed DNA sequences. To insert a gene, we recommend using any sites between (and including) the HindIII and XbaI sites. By doing this you will be able to insert most of our other products upstream, or downstream, of your gene. Click here to read more about the purpose of each restriction 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 genes (other than CAT) to high levels under the CMV promoter when inserted into the main MCS. The start codon of these reporters was positioned within the NcoI site and the stop codon was positioned within the XbaI site. Expression was validated in the A549 lung carcinoma cell line. Expression of CAT is simultaneously controlled by the Ub promoter, and this has been validated in the same cell line and shown to mediate high levels of CAT expression.    

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.
  • 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.
  • 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.
  • Ub promoter – This promoter has been modified to remove BseRI, StuI and SpeI sites.     

Restriction site notes:

  • Bgl2, KpnI, SwaI, AscI and PmeI each cut the vector at two sites positioned to flank the promoter, start codon, origin, Ub CAT expression cassette, and the 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.