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Puromycin Selection Plasmids

OG379 - pSF-CMV-RSV-Puro Ascl

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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-CMV-RSV-Puro AscI

Product Code: OG379

Quantity Provided: 5µg

Size (bp): 5395

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 Rous Sarcoma Virus (RSV) LTR promoter

Purpose: This vector contains a puromycin resistance expression cassette that is designed as an add-on to any of our vectors. The entire RSV puromycin cassette is flanked by AscI sites. The RSV promoter is considerably weaker than CMV. We have a stronger Ubiquitin promoter driven vector (pSF-CMV-Ub-Puro AscI) in the same format if you require higher levels of resistance.

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 of reporters was validated in 293 cells. The RSV Puro cassette has also been validated in the same cell line and shown to induce resistance to puromycin. 

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:

  • 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.
  • The puromycin resistance gene has been modified to remove a StuI site and an EagI site to ensure compatibility with all of our products. These are all conservative changes and do not change the amino acid sequence.   
  • The Rous Sarcoma Virus (RSV) promoter in this vector has been modified to remove an EcoRI site. This does not reduce expression.    

Restriction site notes:

  • Bgl2, KpnI, SwaI, AscI and PmeI each cut the vector at two sites positioned to flank the promoter, start codon, origin, RSV puromycin resistance 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.