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pTXB1 Vector

Catalog # Concentration Size List Price Quantity Your Price
N6707S 200 µg/ml 10 µg $222.00
$199.80
Catalog # Size List Price Your Price
N6707S 10 µg $222.00
$199.80
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

pTXB1 is an E. coli expression vector designed for the in-frame insertion of a target gene into the polylinker upstream of the Mxe intein/chitin binding domain. The fusion protein is bound to chitin beads and the thiol-induced cleavage activity of the intein releases the target protein.

  • View sequence details
  • Used in the IMPACT™ Protein Purification System
  • Recommended for use in intein-mediated protein ligation and C-terminal labeling
  • Double-stranded vector; 6,706 base pairs in length
  • Ampicillin resistance

IMPACT Citations

pTXB1 is an E. coli expression vector in the IMPACT™ Kit (NEB #E6901) (1,2). It is designed for the in-frame insertion of a target gene into the polylinker upstream of the Mxe intein/chitin binding domain (27 kDa) (2,3,4,5). The fusion protein is bound to chitin beads and the thiol-induced cleavage activity of the intein releases the target protein. pTXB vectors are recommended for use in intein-mediated protein ligation and C-terminal labeling (2). This double stranded vector is 6,706 base pairs in length.

DNASU and Addgene are central repositories for plasmid clones and collections that may also be helpful.

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  pTXB1 Vector N6707SVIAL -20 1 x 0.05 ml 0.2 mg/ml
Features
  • The NdeI site in the polylinker contains an ATG sequence for translation initiation. Unique sites are indicated in bold. SalI site is not unique.
  • The SapI site should be used for cloning of the 3´ end of the insert. Use of the SapI site allows cloning of the target protein adjacent to the intein, resulting in cleavage of the target protein without any additional amino acids at its C-terminus.
  • Expression of the fusion gene is under the control of an IPTG-inducible T7 promoter (6).
  • A pBR322 derivative with a ColE1replication origin.
  • Origin of DNA replication from bacteriophage M13, which allows for the production of single-stranded DNA by helper phage superinfection of cells bearing the plasmid (M13K07 helper phage, NEB #N0315).
  • Ampicillin resistance.
  • Other IMPACT vectors are available which allow for fusion of a target gene to N- or C- terminus of an intein. The cleavage reaction may be induced by thiol reagent or temperature/pH shift.
  • Companion vector pTXB3 (NEB #N6708) contains an NcoI site in place of NdeI.
  • A wide range of E. coli host strains: T7 Express Competent E. coli (High Efficiency) (NEB #C2566) or BL21(DE3) Competent E. coli (NEB #C2527) and derivatives.

Properties & Usage

Lac Repressor on Plasmid

0

Affinity Tag

Chitin-Binding Domain (CBD)

Companion Products
References
  • Chong, S., Mersha, F.B., Comb, D.G., Scott, M.E., Landry, D., Vence, L.M., Perler, F.B., Benner, J., Kucera, R.B., Hirvonen, C.A., Pelletier, J.J., Paulus, H. and Xu, M.-Q. (1997). Single-column purification of free recombinant proteins using a self-cleavableaffinity tag derived from a protein splicing element. Gene. 192, 271-281.
  • Evans, T.C., Benner, J. and Xu, M.-Q. (1998). Semisynthesis of cytotoxic proteins using a modified protein splicing element. Protein Sci.. 7, 2256-2264.
  • Southworth, M.W., Amaya, K., Evans, J., T.C., Xu, M.-Q. and Perler, F.B. (1999). Purification of proteins fused to either the amino or carboxy terminus of the Mycobacterium xenopi gyrase A intein.. BioTechniques. 27, 110-120.
  • Watanabe, T., Ito, Y., Yamada, T., Hashimoto, M., Sekine, S. and Tanaka, H. (1994). The role of the C-terminal domain and type III domains of chitinase A1 from Bacillus circulans WL-12 in chitin degradation. J. Bacteriol.. 176, 4465-4472.
  • Telenti, A., Southworth, M., Alcaide, F., Daugelat, S., Jacobs, W.R. Jr. and Perler, F.B. (1997). The Mycobacterium xenopi GyrA protein splicing element: Characterization of a minimal intein. J. Bacteriol.. 179, 6378-6382.
  • Dubendorff, J.W. and Studier, F.W. (1991). Controlling basal expression in an inducible T7 expression system by blocking the target T7 promoter with lac repressor. J. Mol. Biol.. 219, 45-59.
DataCards
Quality Control Assay
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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]
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]
Safety Data Sheets
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