pSNAP-tag® (T7)-2 Vector

Catalog #	Concentration	Size
N9181S	Not Applicable	20 µg
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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.

Product Categories:

Cellular Analysis Vectors Products,

DNA Plasmids & Substrates Products,

Protein Labeling Products

Applications:

Snap Surface Products,

In vivo Imaging,

E Coli Protein Expression
,
Pulse Chase
,
Receptor Internalization
,
Protein Localization
,
Protein Labeling Snap Clip
- Advantages and Features
  
  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^qCompetent E. coli (NEB #C3013). We recommend using T7 Express strains to conduct initial tests of the expression of the fusion gene.
- Properties & Usage
  Materials Required but not Supplied
  
  IPTG inducible E. coli strains: T7 Express
  
  T7 Express l^q Competent E. coli (NEB #C2566)
  
  T7 Express lysY Competent E. coli (NEB #C3016)
  
  T7 Express Competent E. coli (NEB #C3010)
  
  lysY/l^q Competent E. coli (NEB #C3013)
  
  Storage Temperature
  -20°C
  Sequence Files
  Fasta GenBank
- Related Products
  Companion Products
  
  T7 Express Competent E. coli (High Efficiency)
  
  T7 Express lysY Competent E. coli (High Efficiency)
  
  T7 Express lysY/I^q Competent E. coli (High Efficiency)
- Product Notes
  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.
Protocols, Manuals & Usage
- Protocols
  Expression of SNAP-tag Fusions (N9181)
  
  Cloning of SNAP-tag Fusions in pSNAP-tag(T7)-2 (N9181)
- Application Notes
  Labeling of Escherichia coli Expressed SNAPtag Fusion Proteins
- Datacards
  The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.)
  
  N9181Datasheet-Lot0011104
  
  N9181Datasheet-Lot0011207
  
  N9181Datasheet-Lot0011310
  
  N9181Datasheet-Lot0011501
  
  N9181Datasheet-Lot0021610
Tools & Resources
- Selection Charts
  Comparison of SNAP-tag®/CLIP-tag™ Technologies to GFP
  
  SNAP-tag®/CLIP-tag® Cloning Vector and Plasmid Selection Chart
- Web Tools
  DNA Sequences and Maps Tool
FAQs & Troubleshooting
- FAQs
  Cellular Imaging and Analysis FAQs
  
  What is the DNA sequence of this product?
- Troubleshooting
  Labeling with SNAP-tag® Technology Troubleshooting Guide
Citations & Technical Literature
- Citations
  
  Product Citation Tool
  
  Powered by Bioz See more details on Bioz
Quality, Safety & Legal
- Quality Control Assays
  
  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
  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
- Certificate of Analysis
  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
- Safety Data Sheets
  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
- Datacards
  The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.)
  
  N9181Datasheet-Lot0011104
  
  N9181Datasheet-Lot0011207
  
  N9181Datasheet-Lot0011310
  
  N9181Datasheet-Lot0011501
  
  N9181Datasheet-Lot0021610
- Legal and Disclaimers
  
  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 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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pSNAP-tag® (T7)-2 Vector

Features

Materials Required but not Supplied

Storage Temperature

Sequence Files

Companion Products

Product Citation Tool

Licenses

Trademarks

pSNAP-tag® (T7)-2 Vector

Features

Materials Required but not Supplied

Storage Temperature

Sequence Files

Companion Products

Product Citation Tool

Licenses

Trademarks

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