pSF-CMV-NH2-V5-EKT-Nco1 (OG91) N-Terminal V5 Tag Plasmid

Product Description

N terminal V5 peptide tag expression plasmid for adding the V5 epitope to the end of a protein of interest. The tag is adjacent to the NcoI site.

$220.15
Product code: OG91
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Plasmid Information

Product Name: pSF-CMV-NH2-V5-EKT-NcoI

Product Code: OG91

Size (bp): 4280 bp

Bacterial Antibiotic Selection: KanR

Origin and Compatibility: pUC high copy derived from pBR322

Bacterial Copy Number: 500-700 per cell

Promoter: Cytomegalovirus (CMV) immediate early promoter

Plasmid Purpose:

This vector adds a V5 epitope tag to the N-terminus of a protein that is encoded within the multiple cloning site. This tag allows the detection and purification of a tagged protein using antibodies raised against the Influenza V5 epitope. The V5 tag coding sequence is GKPIPNPLLGLDST. There is an enterokinase cleavage site (DDDDK) immediately downstream of the V5 tag that can be used to remove the V5 tag from a purified protein. It cleaves after the lysine residue.

Promoter Expression Level:

This plasmid contains the mammalian CMV promoter to drive gene expression. We have tested all of our mammalian promoters in a range of cell types and CMV is consistently the strongest in those we have studied. However there are many reports of the CMV promoter demonstrating silencing by methylation in long-term culture. For this reason we stock a range of other promoters that are compatible with this plasmid and are available on request.

Vector Sequence Files

Please select the file type you require. For reference most cloning programs will import a .gb (Genbank) file and will show all of the plasmids features automatically when downloaded and imported.

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.

Cloning in a Gene:

This vector has been designed to allow the addition of a peptide tag to the end of a protein of interest using standard cloning techniques.

Multiple Cloning Site Notes:

There is a start codon in the NcoI site can be removed by digestion with KpnI if required. The MCS for gene insertions extends from NotI to XbaI however the tag resides between the NotI and HindIII sites. There are Shine-Dalgarno sequences and KOZAK sequences aligned with the start codon of the peptide tag.

The ClaI to NheI sites have other functions such as adding C-terminal peptide tags second promoters or IRES expression components. The BsgI and BseRI restriction sites cleave within the stop codon in the XbaI site and allow the retrospective fusion of C-terminal peptide tags sequences if the stop codon is placed in this position.

Intellectual Property Status

According to our IP-friendly policy this plasmid is sold free of reach-through rights and can be used to make commercial products. However the plasmid itself (or derivatives) cannot be sold.