Composite

Part:BBa_K4759247

Designed by: Jianghua Chen   Group: iGEM23_Jiangnan-China   (2023-10-08)
Revision as of 10:30, 8 October 2023 by Registry (Talk | contribs)

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T7-RBS1-petH-RBS2-petF(D21F)-linker-GFP1-10

Generally, the method of determining whether the redox partner is suitable is through tedious steps such as the construction of plasmids, heterologous expression, construction of catalytic systems, and detection of conversion rate after catalysis. Therefore, we wanted to find a convenient way to do a quick screening. We used the fluorescent protein sfGFP to successfully construct a sensor to detect redox partners. sfGFP is a superfolder fluorescent protein that emits green light when irradiated with ultraviolet light. What is special about it is that it can be broken into two parts. We divide sfGFP into sfGFP-1-10 and sfGFP-11, and although these two parts are cut off, there is an interaction force between them, and as long as they are properly folded in space, they will emit light again. Thus, four iron redox proteins are fused to the N-terminus of sfGFP-1-10 and Olep to the C-terminus of sfGFP-11, respectively.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1336
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1336
    Illegal NotI site found at 1109
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1336
    Illegal BglII site found at 1647
    Illegal BglII site found at 2316
    Illegal BamHI site found at 1330
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1336
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1336
  • 1000
    COMPATIBLE WITH RFC[1000]


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