Composite

Part:BBa_K1072018

Designed by: Junqi kuang   Group: iGEM13_SCUT   (2013-09-22)

pGAL1 + Flag + Brho + odr-10 + GFP + ADH1Te + pFUS1 + BFP + ADH1Te

BBa_K1072018 is a composite device, including Odr-10 ,GFP and reporter gene, BFP.

BBa_K1072018 encodes a diacetyl-sensing G-protein coupled receptor, 7-transmembrane receptors, which can be functionally expressed in S. cerevisiae. We add Flag tag, Brho signal peptide in C-terminal and link GFP coding region in N-terminal. If cotransforming it into cells, Odr10 located at the plasma membrane and activated the downstream MAPK pathway,figure1.Finally,BFP was detected, meaning that Odr10 work and activate the downstream pathway.

Figure 2. BFP was detected in yeast.



This part is an improvement of BBa_K211002 and BBa_K775003 made by 2009 Hongkong university and 2012 TU delft team, respectively. A flag tag, Brho signal peptide, GFP and reporter divece with FUS1 promotor have been added.


Sequence and Features


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
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2324


[edit]
Categories
//cds/membrane/receptor
//function/cellsignalling
//function/reporter/fluorescence
Parameters
chassisS.cerevisiae
directionForward
familyG protein family
proteins1)Odr-10 2)GFP 3)BFP