Composite

Part:BBa_K1897011:Design

Designed by: Ang Shi Hui   Group: iGEM16_NUS_Singapore   (2016-10-10)


Invasin


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 4
    Illegal XhoI site found at 2791
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 976
    Illegal AgeI site found at 2485
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

There are two HA tags available for characterisation of the protein produced via western blot. Note that the stop codon for Invasin CDS BBa_K1897010 is shifted to after the HA tags.

Source

Invasin is derived from Yersinia pseudotuberculosis, from the coding sequence [Genbank M17448.1] (see BBa_K1897010. The promoter was synthesized based on the sequence obtained from BBa_R0062. The ribosome binding site (RBS) was synthesized based on the sequence obtained from BBa_B0030. The transcription terminators were synthesized based on the sequence obtained from BBa_B0015.

References

Isberg, R. R., & Falkow, S. (1985). A single genetic locus encoded by Yersinia pseudotuberculosis permits invasion of cultured animal cells by Escherichia coli K-12. Nature, 317(6034), 262-264.

Isberg, R. R., & Leong, J. M. (1988). Cultured mammalian cells attach to the invasin protein of Yersinia pseudotuberculosis. Proceedings of the National Academy of Sciences, 85(18), 6682-6686.

McCormick, B. A., Nusrat, A., Parkos, C. A., D'Andrea, L., Hofman, P. M., Carnes, D., Liang, T.W. & Madara, J. L. (1997). Unmasking of intestinal epithelial lateral membrane beta1 integrin consequent to transepithelial neutrophil migration in vitro facilitates inv-mediated invasion by Yersinia pseudotuberculosis. Infection and immunity, 65(4), 1414-1421.