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OG13 - pSF-CMV-SC101 Ori

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Product Code: OG13 

Size (bp): 5706

Bacterial Antibiotic Selection:
 Kanamycin 

Origin and Compatibility:
 SC101 origin very low-copy origin (approximately 5 copies per cell), derived from plasmid pSC101. 

Copy Number:
 ~5 copies per cell 

Promoter:
 Cytomegalovirus (CMV) immediate early promoter 

Purpose:
 Provides a low copy plasmid, which allows greater stability of difficult to clone, unstable, or toxic, or large DNA sequences.

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 reporter genes to high levels under the CMV promoter. 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. The origin has been validated to produce the correct number of copies in E.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:

  • 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. 
  • SC101 Origin – The SC101 origin has been modified to remove an EcoRV site and a BseRI site. These changes do not alter the origins properties.

Restriction site notes:

  • Bgl2, KpnI, SwaI and PmeI each cut the vector at two sites positioned to flank the promoter, start codon, 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. 
  • The SC101 origin contains a PciI site. This site conflicts with certain products, including pSF-CMV-MCS-IRES PciI.