Coding

Part:BBa_K4759058:Design

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


petH-RBS2-petF(D58Y)


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 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

Through previous experimental results, we selected ferredoxin reductase PetH (SEQ ID NO.7) and ferredoxin PetF (SEQ ID NO.8) from the algae (Synechocystis PCC 6803) as the redox chaperones of OleP. The modeling group docked the two proteins, PetF and OleP, to obtain the hot spot residues and the corresponding mutated amino acids, and the petF genes were mutated by PCR to obtain the corresponding mutants. The original base of the amino acid at position 58 encoding petF was changed to TAT.The amino acid at position 58 is changed from Asp to Tyr.


Source

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

References