• My NEB
  • Print
  • PDF
  • pKYB1 Vector

    Discontinued Date

    11/01/2012
    Catalog #SizeConcentrationPriceQtyAdd to Cart
      
    Categories:
    Discontinued Products

    Description

    pKYB1 is an E. coli expression vector for use with the IMPACT™ Kit (NEB #E6901) (1,2). It is designed for in-frame insertion of a target gene into the polylinker, upstream of the Sce VMA intein/chitin binding domain (55 kDa)(1,3). pKYB1 carries the kanamycin resistance gene (Kn) from Tn903. This double stranded vector is 8,393 bp in length.

    Advantages and Features

    Features

    • A pBR322 derivative. 
    • The NdeI site in the polylinker contains an ATG sequence for translation initiation. 
    • Use of the SapI site allows for cloning of the target protein adjacent to the intein, resulting in purification of the target protein without any additional amino acids at its C-terminus. 
    • Unique sites are in bold. NheI and NruI are not unique. 
    • Expression of the fusion gene is under the stringent control of the T7 promoter (4) and is regulated by IPTG due to the presence of a lacI gene. 
    • Origin of DNA replication from bacteriophage M13, which allows for the production of single-stranded DNA by helper phage (M13K07 Helper Phage, NEB #N0315) superinfection of cells bearing the plasmid. 
    • Kanamycin resistant. 
    • Compatible restriction sites for subcloning a fusion gene from other IMPACT vectors. 
    • 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 and Usage

    Affinity Tag

    Other

    Storage Temperature

    -20°C

    Sequence Files

    Fasta  GenBank  

    References

    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, 271-281.
    2. Chong, S., Shao, Y., Paulus, H., Benner, J., Perler F.B. and Xu, M.-Q. (1996). Protein splicing involving the Saccharomyces, cerevisiae VMA intein: the steps in the splicing pathway, side reactions leading to protein cleavage, and establishment of an in vitro splicing system. J. Biol. Chem.. 271, 22159-22168.
    3. 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.
    4. 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.

    FAQs

    1. What is IMPACT?
    2. What vectors are included in the IMPACT kit?

    Interactive Tools

    Application Notes

    Material Safety Datasheets

    The following is a list of Material Safety Data Sheets (MSDS) that apply to this product to help you use it safely. The following file naming structure is used to name these document files: [Product Name] MSDS. For international versions please contact us at info@neb.com.