Part:BBa_K2587027:Design

luxI_luxR_P_lux_gfp

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  Illegal NheI site found at 829
  Illegal NheI site found at 852
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 1082
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 892
  Illegal BsaI site found at 1681
  Illegal BsaI.rc site found at 1523
  Illegal BsaI.rc site found at 1790
  Illegal BsaI.rc site found at 2468
  

Design Notes

This construct was created using the CIDAR MoClo toolbox (1). The toolbox offers different ways of how to assemble multiple parts in a modular way. For instance, if desired, promoters, RBS and/or terminators can be exchanged by combining the different parts. In this way, strength of the promoters or RBS with different properties can be adapted. In this case one of the strongest promoters are used for expression of LuxI and LuxR, as described in the supplementary material of the CIDAR MoClo toolbox. For expression of GFP, the inducible pLux promoter is used. All the used parts are codon optimised for E.coli.

:
Figure 1: Plasmid card showing the control plasmid for functionality of pLux. Each gene is part of its own transciptional unit, comprising promoter, RBS and terminator. This order is dictated by the CIDAR MoClo toolbox and allows modular assembly of parts.GfP expression is induced upon synthesis of AHL and subsequent binding to LuxR and finally to pLux.

Source

The sequences for LuxI, LuxR, pLux and phiX174 E are from GenBank and codon optimised for E.coli. The remaining sequences are from the CIDAR MoClo toolbox.

References

(1) https://www.addgene.org/cloning/moclo/densmore/#kit-contents