pSNAP-tag^® (T7)-2 Vector

Features

BACKGROUND:

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 l^q Competent E. coli (NEB #C3016), T7 Express lysY Competent E. coli (NEB #C3010) and T7 Express lysY/l^q Competent E. coli (NEB #C3013). We recommend using T7 Express strains to conduct initial tests of the expression of the fusion gene.

Companion Products

• T7 Express lysY/I^q Competent E. coli (High Efficiency)
• T7 Express lysY Competent E. coli (High Efficiency)
• T7 Express Competent E. coli (High Efficiency)

1. 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.

1. Keppler, A. et al. (2003). Nat. Biotechnol. 21, 86.
2. Gautier, A. et al. (2008). Chem. Biol. 15, 128.
3. Keppler, A. et al. (2004). Proc. Natl. Acad. Sci. USA. 101, 9955.
4. Dubendorff, J.W. and Studier, F.W. (1991). J. Mol. Biol. 219, 45.

1. Expression of SNAP-tag Fusions (N9181)
2. Cloning of SNAP-tag Fusions in pSNAP-tag(T7)-2 (N9181)

• Labeling of Escherichia coli Expressed SNAP-tag^® Fusion Proteins

• Comparison of SNAP-tag®/CLIP-tag™ Technologies to GFP
• SNAP-tag®/CLIP-tag® Cloning Vector Selection Chart

• DNA Sequences and Maps Tool

1. Cellular Imaging and Analysis FAQs
2. What is the DNA sequence of this product?

• Labeling with SNAP-tag® Technology Troubleshooting Guide

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]

• N9181S_v1

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]

• N9181S_v1_0021804
• N9181S_v1_10011151
• N9181S_v1_10032928
• N9181S_v1_10073582
• N9181S_v1_10128035
• N9181S_v1_10146314

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

• pSNAP-tag^® (T7)-2 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 email us at [email protected].

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.