PSF-CMV-NH2-HIS-EKT-NCOI - N-TERMINAL HIS TAG PLASMID OGS211-5UG

NACRES:

NA.85

This vector adds a His epitope tag to the N-terminus of a protein that is encoded within the multiple cloning site. This tag allows the purification of a tagged protein by binding to metal matrices such as nickel or cobalt. There is an enterokinase cleavage site (DDDDK) immediately downstream of the His tag that can be used to remove the His 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.
Multiple cloning site notes: The his tag 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 that contain an NcoI at the 5 prime. This tag is positioned immediately upstream of the NcoI site to minimise the amount of extra amino acids that are added to the N-terminus of your gene.
The start codon that is within the NcoI restriction site is in frame with and adjacent to the coding sequence for the His/EKT peptide tag. Shine-Dalgarno sequences and KOZAK sequences are positioned correctly at the start of the His signal peptide. The ClaI to NheI sites have other functions such as adding peptide tags or IRES elements. The BsgI and BseRI restriction sites cleave within the stop codon in the XbaI site and allow fusion with our C-terminal peptide tags provided that your genes stop codon is also positioned here.
Sequence
Analysis Note

form	buffered aqueous solution
mol wt	size 6152 bp
conjugate	6-His tagged
Origin of replication	pUC (500 copies)
Peptide cleavage	EKT
Peptide tag location	N-terminal
Promoter	Promoter name: CMV Promoter activity: constitutive Promoter type: mammalian
bacteria selection	kanamycin
reporter gene	none
shipped in	ambient
storage temp.	20°C

RIDADR	NONH for all modes of transport
Flash Point F	Not applicable
Flash Point C	Not applicable