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Blasticidin Selection Plasmids

OG383 - pSF-CMV-RSV-Blast 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-Blast AscI

Product Code: OG383

Quantity Provided: 5µg

Size (bp): 5224

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 blasticidin (blast) resistance gene driven by the RSV promoter that is designed as an add-on to any of our vectors. The entire RSV blast expression 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-Blast AscI) in the same format if you require high levels of reporter gene activity.

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 Blast cassette has also been validated in the same cell line and shown to induce resistance to blasticidin.

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.
  • Blasticidin resistance gene – The blast gene has been modified to remove StuI, BsgI and SacI sites. These changes did alter the amino acid sequence of the protein and do not effect resistance.  
  • 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 Blasticidin 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.