pSNAP_f Vector

Reagents Supplied

The following reagents are supplied with this product:

NEB #	Component Name	Component #	Stored at (°C)	Amount	Concentration
N9183S     -20       pSNAPf Vector N9183SVIAL -20 1 x 0.04 ml 500 µg/ml

Features

BACKGROUND:

A plasmid map and the sequence of the cloning region can be found with these instructions. The complete plasmid sequence can be downloaded. This plasmid encodes the gene SNAP_f which is a mutant form of the human gene for O⁶-alkylguanine-DNA-alkyltransferase (hAGT). The codon usage of the gene is optimized for expression in mammalian cells. In the plasmid sequence, the SNAP_f gene is encoded from 969 bp to 1514 bp.

This plasmid is intended for the cloning and stable or transient expression of SNAP-tag protein fusions in mammalian cells. It is suitable for the efficient production of stable cell lines expressing SNAP_f gene fusions. The plasmid contains the CMV promoter followed by the genes for SNAP_f and neomycin resistance separated by the IRES of the encephalomyocarditis virus (ECMV), which permits the translation of two open reading frames from one messenger RNA; therefore after selection of stable mammalian cells for neomycin resistance, nearly all surviving colonies should stably express the SNAP_f fusion protein. Unless your expression experiments require a pure population of cells, you can simply use the pool of resistant cells, otherwise cell clones can be isolated and characterized using standard procedures.

The plasmid contains the β-lactamase (Ampicillin resistance) gene for maintenance in bacteria. The gene of interest can be cloned upstream or downstream of the SNAP_f coding sequence, as a fusion to the N- or C-terminus of the SNAP-tag. pSNAP_f Vector can also be used as an expression control plasmid, expressing SNAP_f alone, in which case the SNAP-tag protein is distributed throughout the cell. The SNAP_f gene can be isolated from the plasmid using PCR or direct cloning in order to subclone it into a different vector of choice.

1. Storage: pSNAP_f Vector issupplied in TE buffer (10 mM Tris-HCl, pH 8.0, 1 mM EDTA) at a concentration of0.5 µg/µl. Plasmid solutions can be stored at 4°C for up to one week. Forlong-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. Maurel, D. et al. (2008). Nat. Methods. 5, 561.
5. Jansen, L.E. et al. (2007). J. of Cell Biol. 176, 795.
6. Krayl, M., Guiard, B. Paal, K. and Vous, W. (2006). Anal. Biol. Chem. 355, 81-89.
7. Banala, S., Arnold, A. and Johnsson, K. (2008). ChemBio Chem. 9, 38-41.

1. Expression of SNAP_f Fusions (N9183)
2. Cloning of SNAP-tag Fusions in pSNAP_f (N9183)

• Simultaneous Fluorescent Labeling of Proteins in Living Cells

• 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]

• N9183S_v2

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]

• N9183S_v2_10013726
• N9183S_v2_10034364
• N9183S_v2_10068402
• N9183S_v2_10112792
• N9183S_v2_10178398

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

• pSNAP_f Vector

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

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.

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.

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 CMV promoter is covered under U.S. Patent No. 5,385,839 and its use is permitted for research purposes only. Any other use of the CMV promoter requires a license from the University of Iowa Research Foundation, 214 Technology Innovation Center, Iowa City, IA 52242.

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).