Coding

Part:BBa_K4759095:Design

Designed by: Junyuan Shi   Group: iGEM23_Jiangnan-China   (2023-10-02)


petH-RBS2-petF(D67Y)-linker-GFP1-10


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1249
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1249
    Illegal NotI site found at 1022
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1249
    Illegal BglII site found at 1560
    Illegal BglII site found at 2229
    Illegal BamHI site found at 1243
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1249
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1249
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

To rapidly screen the most compatible redox chaperone with Olep, a novel redox chaperone sensor was constructed to measure the interaction force between Olep and the redox chaperone based on bimolecular fluorescence complementation techniques. The sensor uses a superfolded green fluorescent protein (sfGFP), and sfGFP is divided into sfGFP-1-10 and sfGFP-11 without fluorescence. The interaction between sfGFP-1-10 and sfGFP-11 pulls down the distance in space. We performed site-directed mutagenesis of petF (D67Y)and ligated GFP1-10 at the C terminus of PetF.


Source

ferredoxin reductase PetH (SEQ ID NO.7) and ferredoxin PetF (SEQ ID NO.8) is from the algae (Synechocystis PCC 6803)


References