Composite

Part:BBa_K1073032

Designed by: iGEM Team Braunschweig 2013   Group: iGEM13_Braunschweig   (2013-09-17)
Revision as of 07:32, 25 September 2013 by Laura S (Talk | contribs)

Autoinducer synthase LasI + LVA with combined eforRed chromoprotein expression cassette

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

Usage and Biology

This device can be used to produce the autoinducer N-3-oxododecanoyl-HSL of the las qourum sensing system of Pseudomonas aeruginosa. N-3-oxododecanoyl-HSL builds a complex with the transcription regulator LasR. 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 in front of lasI it can be combined with various regulatory regions (e.g. a constitutively or regulated promoter) depending on intended use.

The constitutively expressed chromoprotein eforRed is added to the construct and can be used as a selection marker. EforRed exhibits a pink color when expressed and can be detected in less than 24 h during cultivation on agar plates or liquid culture (see BBa_K592012).



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 838
    Illegal NheI site found at 861
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 680
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 242
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None