Composite

Part:BBa_K1132020:Design

Designed by: iGEM Toulouse   Group: iGEM13_INSA_Toulouse   (2013-09-20)

rbs-YF1-rbs-FixJ-term-pFixK2-mRFP1


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 NgoMIV site found at 562
    Illegal NgoMIV site found at 634
    Illegal NgoMIV site found at 724
    Illegal NgoMIV site found at 742
    Illegal NgoMIV site found at 1254
    Illegal NgoMIV site found at 1547
    Illegal NgoMIV site found at 1641
    Illegal AgeI site found at 276
    Illegal AgeI site found at 1422
    Illegal AgeI site found at 2686
    Illegal AgeI site found at 2798
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1311
    Illegal BsaI.rc site found at 175


Design Notes

This composite part can be used for a blue light sensitive system. It is built with 4 preexisting parts : rbs-YF1-rbs-FixJ (BBa_K592016) with a terminator (BBa_B1010) and followed by FixK2 (BBa_K081014)-rbs-mRFP1-term (BBa_K592006). Rbs-YF1-rbs-FixJ is a regulatory protein working with the promotor FixK2. In darkness, YF1 is phosphorylated and gives its phosphate group to FixJ. Once phosphorylated, FixJ can bind to FixK2 promoter and actives it. In light, FixK2 promoter can't work.
A final construction was made with a constitutive promoter (Bba_J23116) but this promoter seems to weak to generate enought copies of YF1-FixJ to activate FixK2.
Adding a strong promoter before this biobrick can improve the system.


Source

BBa_K592016
BBa_B1010
BBa_K081014
BBa_K592006

References