Composite

Part:BBa_K1073031

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

Autoinducer synthase LuxI + LVA with combined amilGFP chromoprotein expression cassette

This part builds the basis for the construction of an expression cassette of the autoinducer synthase LasI which synthesizes N-3-oxohexanoyl-homoserine lactone. It is combined with a expression cassette of the chromoprotein amilGFP. The device lacks a promoter upstream of luxI and therefore only leads to expression of amilGFP.


Usage and Biology

This device can be used to produce the autoinducer N-3-oxohexanoyl-HSL of the lux qourum sensing system of Vibrio fischeri. N-3-oxohexanoyl-HSL builds a complex with the transcription regulator LuxR. This complex then binds to specific sequences in the las promoter region activating the expression of genes located downstream (not included in this device). Since this contruct does not include a promoter upstream of luxI it can be combined with various regulatory regions (e.g. constitutively or regulated promoter) depending on intended use.

The constitutively expressed chromoprotein amilGFP is added to the construct and can be used as a selection marker. AmilGFP exhibits a yellow color when expressed and can be detected in less than 24 h during cultivation on agar plates or in liquid culture (see BBa_K592010).


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 814
    Illegal NheI site found at 837
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 656
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
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