Composite

Part:BBa_K4059051:Design

Designed by: Julie le-Fevre Hansen   Group: iGEM21_SDU-Denmark   (2021-10-21)


CPR from Phanerochaete chrysosporium with N-terminal substitution (5), E. coli K12 codon optimized,


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal NotI site found at 1516
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The gene encodes a NADPH-cytochrome P450 reductase (CPR) delivers the electrons necessary for the reaction carried out by the 4-monooxygenase (PsiH). This gene originates from the mushroom Phanerochaete chrysosporium. CPR is like PsiH usually bound to the membrane of the endoplasmatic reticulum by the N-terminal, which is strongly hydrophobic. When expressed in bacteria that lack the endoplasmatic reticulum, the N-terminal can affect the folding of the enzyme leading to an ineffective protein. Therefore N-terminal modifications are needed. This brick includes a modification in form of a substitution of an entire N-terminal from another enzyme, which is soluble within the cell.

Source

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References