Composite

Part:BBa_K1462580:Design

Designed by: zhanru Chen   Group: iGEM14_SCUT   (2014-10-14)

GAL1+GBD+SH3+SH3+PDZ+PDZ+Tom22+ADH1


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 2082
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 2082
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 615
    Illegal BamHI site found at 648
    Illegal BamHI site found at 820
    Illegal BamHI site found at 1029
    Illegal BamHI site found at 1238
    Illegal BamHI site found at 1561
    Illegal BamHI site found at 1884
    Illegal XhoI site found at 801
    Illegal XhoI site found at 1010
    Illegal XhoI site found at 1219
    Illegal XhoI site found at 1542
    Illegal XhoI site found at 1865
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 2082
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 2082
    Illegal AgeI site found at 150
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Scaffold protein, with a leading peptide Tom22, can locate on the outer mitochondria membrane with the scale of GBD: SH3: PDZ==1:2:2.We built up the scale to affirm the best scale. In this way , we tried to fix the PRK, RuBisCO, CA to a limited space to make full use of ATP and CO2 so that we coule improve the pyruvate offering to the butanol pathway

Figure 1.This is the pathway and the structure of the carbon capture. As we see, with the help of ATP,the RuBiSco-5P change into RuBiSco-1,5P by PRK, and then change into 3P-glyrerate by RuBiSco, and CA can help us to adjust the concentration of carbon dioxide. The x,y,z are the number of scaffold unit.

Source

The GBD,SH3,PDZ domain are synthesis from Genewiz, the Tom22 is from the PCR with the template S.cerevisiae genome.GAL1 and ADH1 is from the plate of IGEM.

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] 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).