Composite

Part:BBa_K1073030

Designed by: iGEM Team Braunschweig 2013   Group: iGEM13_Braunschweig   (2013-09-17)

Inducible ampicillin resistance cassette + LasR expression cassette


Induction of the beta lactamase mediated ampicillin resistance by LasR and specific autoinducer N-3-oxododecanoyl homoserine lactone.

In complex with the autoinducer molecule N-3-oxododecanoyl homoserine lactone (N-3-oxododecanoyl-HSL) the transcription regulator LasR binds to specific sequences in the promoter region and enables transcription of ampR.


Usage and Biology

The expression rate of the genes downstream of the promoter region can be tuned by the concentration of N-3-oxododecanoyl-HSL in the media. In this case the expression of beta lactamase can be induced by adding N-3-oxododecanoyl-HSL to the culture broth. Expression of lasR is regulated by the strong constitutive promoter BBa_J23100 and RBS of medium strengh (BBa_B0032).

Note that due to the leakiness of the promoter there is background activity of the beta lactamase even when N-3-oxododecanoyl-HSL is not present.



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1169
    Illegal NheI site found at 1192
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1546
    Illegal AgeI site found at 1743
  • 1000
    COMPATIBLE WITH RFC[1000]


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Parameters
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