Composite

Part:BBa_K4195118

Designed by: Honglin Song   Group: iGEM22_XMU-China   (2022-09-28)
Revision as of 07:44, 13 October 2022 by CZL (Talk | contribs) (→‎Biology)

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I0500-B0034-araC-B0034-GFP-B0015

I0500-B0034-araC-B0034-GFP-B0015 I0500 is the inducible pBad/araC promoter. This composite part can express GFP induced by arabinose, which is used to detect the constitutive expression of the system.


Usage

pBad is an E. coli promoter that is tightly controlled by: inducer: L-arabinose. repressor: AraC acts as the repressor one study concluded that arabinose can change the conformation of araC and prevent it from successfully binding to and repressing pBad. We use BBa_K4195018 and BBa_K2560043 to construct the expression system and obtained the composite part BBa_K4195118. The expression level of GFP can be used to detect the constitutive expression of the system.

Biology

AraC In the absence of arabinose, the AraC dimer adopts a different conformation and binds to araO2 and araI1. In this position, there is no monomer at site araI2, and so the protein cannot activate the araBAD promoter (araPBAD). When arabinose is present, arabinose binds to AraC, changing the shape of that activator so that it binds as a dimer to araI1 and araI2. This places one monomer of AraC close to the promoter from which it can activate transcription.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1144
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 979
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2785
    Illegal SapI site found at 961


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Categories
Parameters
None