pSNAP-tag® (T7)-2 Vector is an Escherichia coli expression plasmid encoding the SNAP-tag protein. Expression is under control of the IPTG inducible T7 promoter.
This plasmid is intended as an expression vector for SNAP-tag gene fusions in appropriate E. coli strains
The E. coli strain used for expression needs to provide T7 polymerase in order to achieve expression from the T7 promoter upstream of the gene
The vector can be used for expression of the SNAP-tag protein alone (20 kDa)
Gene of interest can be fused to the N-terminus or C-terminus of SNAP-tag
Product Information
pSNAP-tag (T7)-2 Vector is an Escherichia coli expression plasmid encoding the SNAP-tag protein. Expression is under control of the IPTG inducible T7 promoter. The pSNAP-tag (T7)-2 Vector can also be used as a control plasmid expressing the SNAP-tag protein (20 kDa). The target gene should be cloned as a fusion to the N- or C-terminus of the SNAP-tag.
The SNAP-tag is a novel tool for protein research, allowing the specific, covalent attachment of virtually any molecule to a protein of interest. The SNAP-tag is a protein based on human O6-alkylguanine-DNA-alkyltransferase (hAGT)(1,2,3). SNAP-tag substrates are derivatives of benzyl purines and benzyl chloropyrimidines. In the labeling reaction, the substituted benzyl group of the substrate is covalently attached to the SNAP-tag.
There are two steps to using this system: sub cloning and expression of the protein of interest as a SNAP-tag fusion, and labeling of the fusion with the SNAP-tag substrate of choice. Cloning and expression of SNAP-tag fusion proteins are described in this document. The labeling of the fusion proteins with SNAP-tag substrates is described in the instructions supplied with SNAP-tag substrates.
Cloning Region of pSNAP-tag® (T7)-2 Vector
Unique restriction sites in the regions flanking the SNAP26b gene
are displayed above the coding strand.
This product is related to the following categories:
This plasmid encodes the gene SNAP26b which is a mutant form of the human gene for O6-alkylguanine-DNA-alkyltransferase (hAGT). (Compared to wild type hAGT, the SNAP-tag protein contains the mutations C26A, K125A, A127T, R128A, G131K, G132T, M134L, R135S, C150S, N157G, S159E, and is truncated after G182). The codon usage of the gene is optimized for expression in E. coli. In the plasmid sequence, the SNAP26b gene is encoded from bp 5767 to 6311. It is not flanked by additional tags for purification. The plasmid contains the pMB1 origin of replication from pBR322 and is maintained at a similar copy number to pBR322 (at a moderate to low copy number of 20 copies/cell). It encodes an ampicillin resistance gene.
This plasmid is intended as an expression vector for SNAP-tag gene fusions in appropriate E. coli strains. The strain needs to provide T7 polymerase in order to achieve expression from the T7 promoter upstream of the gene (4). Expression of the SNAP-tag in pSNAP-tag® (T7)-2 Vector has been tested in the following E. coli strains supplied by NEB: T7 Express Competent E. coli (NEB #C2566), T7 Express lq Competent E. coli (NEB #C3016), T7 Express lysY Competent E. coli (NEB #C3010) and T7 Express lysY/lqCompetent E. coli (NEB #C3013). We recommend using T7 Express strains to conduct initial tests of the expression of the fusion gene.
Storage: pSNAP-tag® (T7)-2 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.
References
Keppler, A. et al. (2003). Nat. Biotechnol. 21, 86.
Gautier, A. et al. (2008). Chem. Biol. 15, 128.
Keppler, A. et al. (2004). Proc. Natl. Acad. Sci. USA. 101, 9955.
Dubendorff, J.W. and Studier, F.W. (1991). J. Mol. Biol. 219, 45.
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.
Specifications & Change Notifications
The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@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.
New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.
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 www.neb.com/cia/legal.
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.
The products and/or their use may be covered by one or more of the following patents and patent applications: Methods for Using O6-Alkylguanine-DNA-Alkyltransferases: US 7,939,284; US 8,367,361; EP 1 410 023; EP 2 037 271; EP 1 696 234; EP 2 211 177; JP 4195815; JP 4226053; CN 1295510; CN 1975422; and SG 100125.
Mutants of O6-Alkylguanine-DNA-Alkyltransferases: US 7,888,090. EP 1720981 (pending).
Notice To Buyer/User: The Buyer/User has a non-exclusive license to use this system or any component thereof for RESEARCH AND DEVELOPMENT PURPOSES ONLY. Commercial use of this system or any components thereof requires a license from New England Biolabs, Inc., 240 County Road, Ipswich, MA 01938.
Trademarks
NEW ENGLAND BIOLABS® and SNAP-TAG® are registered trademarks owned by New England Biolabs, Inc.
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