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Ha Tag Plasmids



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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-HA-EKT1

Product Code: OG324

Quantity Provided: 5µg

Size (bp): 4298

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 HA 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 HA epitope. The HA tag coding sequence is YPYDVPDYA. There is an enterokinase cleavage site (DDDDK) immediately downstream of the HA tag that can be used to remove the HA tag from a purified protein. It cleaves after the lysine residue.

Frame:  The tag coding sequence is in frame with the ATG start codon that is within the NcoI restriction site, allowing fusion with any genes that we sell in the MCS. We also sell the same tag much closer to the NcoI site (called pSF-CMV-HA-EKT-NcoI).   

Quality Validation: This vector has been demonstrated to express reporter genes to high levels under the CMV promoter with the HA tag fused to the N-terminus of the reporter gene coding sequence. The start codon of these reporters was positioned at the start of the HA tag and the stop codon was positioned within the XbaI restriction site. The presence of the peptide increased the activity of the reporter gene by 0.7-fold. Reporter gene expression was validated in the A549 lung carcinoma cell line.   

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 SnapFastTM 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.