Composite

Part:BBa_K1072012

Designed by: Junqi kuang, Yaqing du and Junjie zhang   Group: iGEM13_SCUT   (2013-09-08)

Pgpa1+Chimeric Gpa1(Gα subunit)+RFP+ADH1

In yeast, as in mammalian cells, the pheromone receptors are coupled to a heterotrimeric G protein. This is composed of Gpa1p (Gα), Ste4p (Gβ) and Ste18p (Gγ) subunits. In order to coupling heterologous GPCRs in yeast, “Transplants” have the five C-terminal residues,originating from a odr3 G-protein of C. elegans, replaced with the corresponding mammalian Gα residues.

A reporter, RFP, and ADH1 terminator have been added to the tail of BBa_K1072011;

Figure 1. Comperison between endogenous GPCR pathway in yeast and motified Odr10 pathway.


Usage and Biology

Before transforming BBa_K1072012 into yeast cells, it is necessary to knockout endogenous gpa1 gene(Gα subunit).

Sequence and Feature


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 2980
    Illegal AgeI site found at 3092
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 227
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Categories
Parameters
None