Composite

Part:BBa_K1462520:Design

Designed by: zhanru Chen   Group: iGEM14_SCUT   (2014-10-10)

GAL1+YFP+SH3-lig+ADH1+GAL1+GFP+PDZ-lig+ADH1+GAL1+BFP+GBD-lig+ADH1


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 150
    Illegal AgeI site found at 1755
    Illegal AgeI site found at 3300
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2804


Design Notes

This part is the final device including the BGY fluorescent protein with the ligand behind. We planned to transmit this device to yeast with the different scale group of GBD,PDZ and SH3 scaffold protein. And we used the confocal to make sure whether the scaffold protein work and the enzyme can be bind on the site of it.


Source

This device is composed of three parts "GAL1 + BFP + GBD ligand + ADH1"[BBa_K1462490],"GAL1 + GFP + BFP ligand + ADH1"[BBa_K1462500] and GAL1 + YFP + SH3 ligand + ADH1[BBa_K1462510].

References

[1] John E Dueber, Gabriel C Wu, G Reza Malmirchegini, et al.Synthetic protein scaffolds provide modular control over metabolic flux.Nat Biotechnol. 2009 Aug;27(8):753-9.
[2] Schultz, J. et al. Specific interactions between the syntrophin PDZ domain and voltage-gated sodium channels. Nat Struct Biol 5, 19-24 (1998).
[3] Kim, A.S., Kakalis, L.T., Abdul-Manan, N., Liu, G.A. & Rosen, M.K. Autoinhibition and activation mechanisms of the Wiskott-Aldrich syndrome protein. Nature 404, 151-158 (2000).
[4] Wu, X. et al. Structural basis for the specific interaction of lysine-containing proline-rich peptides with the N-terminal SH3 domain of c-Crk. Structure 3, 215-226 (1995).
[5] Harris, B.Z., Hillier, B.J. & Lim, W.A. Energetic determinants of internal motif recognition by PDZ domains. Biochemistry 40, 5921-5930 (2001).
[6] Dueber, J.E., Yeh, B.J., Chak, K. & Lim, W.A. Reprogramming control of an allosteric signaling switch through modular recombination. Science 301, 1904-1908 (2003).
[7] Nguyen, J.T., Turck, C.W., Cohen, F.E., Zuckermann, R.N. & Lim, W.A. Exploiting the basis of proline recognition by SH3 and WW domains: design of N-substituted inhibitors. Science 282, 2088-2092 (1998).