Description_x000D_ General description_x000D_ 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._x000D_ 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._x000D_ Application_x000D_ 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._x000D_ 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 sIt`s have other functions such as adding peptide tags or IRES elements. The BsgI and BseRI restriction sIt`s 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._x000D_ Sequence_x000D_ Quick-reference Plasmid Map_x000D_ 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._x000D_ Genebank Vector Sequence File_x000D_ FASTA Vector Sequence File_x000D_ Full Plasmid Map_x000D_ Analysis Note_x000D_ To view the Certificate of Analysis for this product, please visit www.oxfordgenetics.com._x000D_ Other Notes_x000D_ Looking for more vector options to move your experiments forward faster? Consider a custom cloning vector designed and built by Oxford Genetics™. Find out more at Oxford Genetics - Sigma's partner for cloning and expression vectors for molecular biology and synthetic biology applications._x000D_ Legal Information_x000D_ Oxford Genetics is a trademark of Oxford Genetics Ltd