Features

BACKGROUND:

This plasmidencodes the gene MCP which is a mutant carrierprotein. In the plasmid sequence, the MCP gene isencoded from bp 690 to 923. The MCP gene wascloned via HindIII and PstI including additionalflanking restriction sites.

This plasmid is intended as a transient expressionvector for MCP-tag gene fusions in mammalian celllines. It is not recommended for the productionof stable cell lines. The plasmid contains theb-lactamase (Ampicillin resistance) gene formaintenance in bacteria. The plasmid also containsa CMV promoter and SV40 polyadenylation signalsfor correct mRNA processing. The gene of interestshould be cloned downstream of the MCP-tagcoding sequence, as a fusion to the C-terminusof the MCP-tag. An appropriate cell surface signalpeptide should be cloned upstream of the MCP-tagas an N-terminal fusion. A Kozak sequence islocated upstream of the MCP gene to increase thetranslation efficiency of the fusion protein. TheMCP-tag gene can be isolated from the plasmidusing PCR or direct cloning in order to subclone itinto a different vector system of choice.

Materials Required but not Supplied

• Mammalian cell lines 
• Transfection reagents 
• CoA substrates

Storage Temperature

-20°C

Sequence Files

Fasta GenBank

Companion Products

• NEB® 10-beta Competent E. coli (High Efficiency)

1. Storage: pMCP-tag(m) Vector is supplied in TE buffer (10 mM Tris-HCl, pH 8.0, 1 mM EDTA) at a concentration of 0.5 µg/µl. Plasmid solutions can be stored at 4°C for up to one week. For long-term storage -20°C is recommended.
2. NEB 10-beta Competent E. coli (High Efficiency) (NEB #C3019) is recommended for propagating and subcloning this vector. 

1. What is the ACP-tag?
2. How does it work?
3. How specific is the binding of substrate to the ACP-tag?
4. What linker type and length would you recommend?
5. Can I clone my protein as fusion to the N- or C-terminus of the ACP-tag?
6. Are ACP-tag substrates stable to fixation?
7. Can ACP-tag be multiplexed with other protein labeling systems (GFP, Antibody)?
8. Can you use ACP-tag for in vivo FRET?
9. Does the ACP-tag labeling reaction work in Yeast?

1. Expression of MCP Fusions (N9317)
2. Cloning of MCP-tag Fusions in pMCP-tag(m) Vector (N9317)

• SNAP-tag®/CLIP-tag® Cloning Vector and Plasmid Selection Chart

• DNA Sequences and Maps Tool

• NEBioCalculator

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

• pMCP-tag(m) Vector

This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).

While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.

For more information about commercial rights, please contact NEB's Global Business Development team at gbd@neb.com.

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

Licenses

Cellular Imaging and Analysis (i.e., SNAP and CLIP products)

Notice to Buyer/User: The Buyer/User has a non-exclusive license to use this system or any component thereof for RESEARCH AND DEVELOPMENT ONLY. Commercial use of this system or any components thereof requires a license from New England Biolabs, Inc., 240 County Road, Ipswich, MA 01938. For detailed information see our Terms of Use .

The products and/or their use may be covered by one or more of the following patents and patent applications:
7,939,284 (Methods for Using O6-Alkylguanine-DNA-Alkyltransferases)
7,888,090 (Mutants of O6-Alkylguanine-DNA-Alkyltransferases)
8,163,479 Specific Substrates for  O6-Alkylguanine-DNA-Alkyltransferases)
8,178,314 (Pyrimidines Reacting With O6-Alkylguanine-DNA-Alkyltransferases)
PCT/EP2007/057597 (Labeling of Fusion Proteins with Synthetic Probes)
EP07117800 (Drug Delivery)
EP07117802 (Drug Delivery)
EP07120288 (GTPase-Transient Protein Protein Interactions)
These patents and patent applications are owned by Covalys, or owned by the Ecole Polytechnique Fédérale de Lausanne (EPFL) and exclusively licensed to Covalys and NEB.