PSF-CMV-PURO-NH2-INSULINSP-FLAG®-EKT - INSULIN SECRETION AND FLAG® TAG PLASMID OGS1404-5UG

NACRES:

NA.85

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 Human Insulin (iSP) secretory signal peptide (SP) to allow proteins to be exported from the cytosol. During translocation from the cytosol the signal peptide is removed from the protein by endogenous proteases. This plasmid also contains a secondary FLAG protein tag. The sequence of this tag is: DYKDDDDKWe 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 EKT cleavage tag. The protein sequence of the cleavage tag is: DDDDK. Enterokinase (EKT) protease cleaves after the Lysine residue. It can cleave at other basic residues but this is dependent on protein confirmation. If a proline follows the site it will not cut. None of our products contain a proline after the site.
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.
Sequence
Analysis Note
FLAG is a registered trademark of Sigma-Aldrich Co. LLC

form	buffered aqueous solution
mol wt	size 6254 bp
conjugate	FLAG® 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
Secretion signal	human Insulin
bacteria selection	kanamycin
mammalian cells selection	puromycin
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