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His Tag Plasmids

OG331 - pSF-CMV-NH2-His-EKT2


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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-NH2-His-EKT2

Product Code: OG331

Quantity Provided: 5µg

Size (bp): 4290

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

Purpose: This vector adds a His tag to the N-terminus of a protein that is encoded within the multiple cloning site. The His tag coding sequence contains six histidine residues that is upstream of an enterokinase cleavage site (DDDDK) that can be used to remove the tag from a purified protein if required. It cleaves after the lysine residue. The His tag allows the purification of a tagged protein by binding to metal matrices such as nickel or cobalt. The tag can also be used to allow the detection of tagged proteins using antibodies raised against the His tag.

Frame: The tag coding sequence has been placed one base out of frame in relation to the ATG start codon that is within the NcoI restriction site by making a single base pair deletion.      

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.

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.