pTYB1 Vector

  • This product was discontinued on 11/01/2012
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pTYB1 is an E. coli cloning and expression vector (7477 bp) used in the IMPACT™ Kit (NEB #E6901) which allows the overexpression of a target protein as a fusion to a self-cleavable affinity tag (1). This C-terminal fusion vector is designed for the in-frame insertion of a target gene into a polylinker upstream of an intein tag (the Sce VMA intein/chitin binding domain, 55 kDa) (1,2). This results in the fusion of the C-terminus of the target protein to the N-terminus of the intein tag. Thiol-induced self-cleavage of the intein releases the target protein from the chitin-bound intein tag, resulting in a single column purification of the target protein (1).

For fusion of the N-terminus of the target protein to the intein tag, use pTYB21 (NEB #N6709 ) or pTYB22 (NEB #N6710 ).

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

Advantages and Features


  • The NdeI site in the polylinker contains an ATG sequence for translation initiation.
  • The SapI site is used for cloning the 3´ end of the target gene. This places the C-terminus of the target protein immediately adjacent to the intein cleavage site and results in the purification of a target protein without any extra vector-derived residues at its C-terminus.
    Note: The SapI site is not regenerated after cloning.
  • Expression of the fusion gene is under the control of an IPTG-inducible T7 promoter (3).
  • Expression requires an E. coli host that carries the T7 RNA Polymerase gene [e.g., T7 Express Competent E. coli (High Efficiency NEB #C2566), BL21(DE3) Competent E. coli (NEB #C2527) and derivatives].
  • A pBR322 derivative with a ColE1 replication origin.
  • Ampicillin resistance.
  • Origin of DNA replication from the bacteriophage M13 allows for the production of single-stranded DNA by helper phage superinfection of cells bearing the plasmid (M13K07 Helper Phage, NEB #N0315).
  • Companion vectors (pTYB2, pTYB3, pTYB4) differ only in the sites present in the polylinker.
  • 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.

Properties and Usage

Affinity Tag

Chitin-Binding Domain (CBD)

Storage Temperature



  1. 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-cleavable affinity tag derived from a protein splicing element. Gene. 192, 277-281.
  2. 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.
  3. 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. 219, 45-59.


  1. What vectors are included in the IMPACT kit?

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Safety Data Sheet

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