pCLIPf Vector is a mammalian expression plasmid intended for the cloning and stable or transient expression of CLIP-tag protein fusions in mammalian cells. This plasmid encodes CLIPf, a CLIP-tag protein, which is expressed under control of the CMV promoter. The expression vector has an IRES (internal ribosome entry site) and a neomycin resistance gene downstream of the CLIPf for the efficient selection of stable transfectants. pCLIPf Vector contains two multiple cloning sites to allow cloning of the fusion partner as a fusion to the N- or C-terminus of the CLIPf.The CLIP-tag is a novel tool for protein research, allowing the specific, covalent attachment of virtually any molecule to a protein of interest. The CLIP-tag is a small protein based on human O6-alkylguanine-DNA-alkyltransferase (hAGT). CLIP-tag substrates are derivatives of benzylcytosine (BC). In the labeling reaction, the substituted benzyl group of the substrate is covalently attached to the reactive cysteine of CLIP-tag forming a stable thioether link.pCLIPf contains an improved version of CLIP-tag, termed CLIPf. CLIPf displays faster kinetics in in vitro labeling and fast, specific and efficient labeling in live and fixed cell applications, thereby rendering it a desired research tool for analysis of protein dynamics.Although CLIP-tag is based on the same protein as SNAP-tag®, the benzylcytosine substrates form a separate class of substrates, different from the benzylguanine substrates recognized by SNAP-tag. CLIP-tag and SNAP-tag can be used for orthogonal simultaneous labeling.
There are two steps to using this system: sub cloning and expression of the protein of interest as a CLIPf fusion, and labeling of the fusion with the CLIP-tag substrate of choice. Cloning and expression of CLIPf fusion proteins are described in this document. The labeling of the fusion proteins with CLIP-tag substrates is described in the instructions supplied with the CLIP-tag substrates.
Cloning Region of pCLIPfUnique restriction sites in the regions flanking the CLIPf gene are displayed above the coding strand. The complete sequence for pCLIPf and the control plasmids can be downloaded here .
This product is related to the following categories:
Cellular Analysis Vectors Products,
DNA Plasmids & Substrates Products,
Protein Labeling Products
This product can be used in the following applications:
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