Coding

Part:BBa_K3384315

Designed by: Yiqing Zhang   Group: iGEM20_NJTech_China   (2020-10-26)
Revision as of 15:48, 26 October 2020 by Root (Talk | contribs)


Ste5ΔN-CTM

Ste5ΔN-CTM is a highly active Ste5 mutant which can bind to the plasma membrane in the absence of pheromone. Ste5 is an important scaffold protein in the mating pheromone signaling pathway. The 214 amino acids at the N-terminus of Ste5 contain membrane binding sites and mediate the membrane localization of Ste5 under the control of Gβγ. By deleting the N-terminal sequence, the activation of Ste5 is independent of G protein. The C-terminal transmembrane domain of Snc2 was added to the Ste5ΔN C-terminus in order to constitutively recruit Ste5 to the plasma membrane1. As shown in figure 1, the Ste5ΔN-CTM protein can activate the MAPK pathway in the absence of pheromone and promote the expression of pheromone-responsive genes.


File:T--NJTech China--Ste5ΔN-CTM1.png
Fig.1 The results of the homology and structural modelling protein amajLime.


Reference

1. Lamson, R. E., Takahashi, S., Winters, M. J., and Pryciak, P. M. (2006) Dual role for membrane localization in yeast MAP kinase cascade activation and its contribution to signaling fidelity, Curr Biol 16, 618-623.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 365
    Illegal SpeI site found at 1620
    Illegal SpeI site found at 2166
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 365
    Illegal SpeI site found at 1620
    Illegal SpeI site found at 2166
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 365
    Illegal BglII site found at 813
    Illegal BglII site found at 2121
    Illegal XhoI site found at 2058
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 365
    Illegal SpeI site found at 1620
    Illegal SpeI site found at 2166
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 365
    Illegal SpeI site found at 1620
    Illegal SpeI site found at 2166
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
chassisSaccharomyces cerevisiae