This plasmid is designed to express tagged proteins in mammalian cells either by transient transfection or by creating stable cell lines. It contains a puromycin resistance expression cassette using the human Ubiquitin promoter to drive expression and allow for the selection of cells containing the plasmid.
About the Peptide Tag:This plasmid contains an n-terminal C-Myc epitope tag that can be fused to a gene of interest to allow protein detection and/or purification. The sequence of the tag is: EQKLISEEDL. This plasmid also contains a secondary Hexa-Histidine (6His) protein tag. The sequence of this tag is: HHHHHHWe provide a range of dual peptide tag plasmids. This is because some peptide tags provide specific biological properties (e. g., small molecule affinity new epitopes solubility or protein secretion) that are not provided by others.
About the Cleavage Tag: This plasmid also encodes a protease cleavage site that is designed to be positioned between your gene of interest and the tag to allow the removal of the tag following protein purification or isolation. This plasmid contains a Thrombin cleavage tag. The protein sequence of the cleavage tag is: LVPRGS. It cleaves preferentially between the Arg and Gly residues. Off target cleavage can often occur at non-specific sites normally from other contaminating proteases. To ensure maximal protein integrity the enzyme reagent must be highly pure.
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.
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.
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