Composite

Part:BBa_K3151028

Designed by: Tasneem Khandaker   Group: iGEM19_Macquarie_Australia   (2019-10-16)


Lac Cyclic-Di-GMP Riboswitch w/ eGFP Reporter


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 224
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 206


Overview

The expression of green fluorescent protein (GFP) relies on the action of the phosphodiesterase which decreases the intracellular concentration of cyclic-di-GMP available. This decrease in intracellular c-di-GMP then activates the riboswitch (Hengge, R. 2009). Our riboswitch plasmid contains a Plac promoter, the riboswitch, and a terminator. These three components form the riboswitch composite part. Combined, these catalyse the activation of the riboswitch, and consequently the production of green fluorescent protein.



Assembly and Design

Suitable inducible promoters were identified from the Parts Registry to be used in the part characterisation for the riboswitch. The Plac promoter was assembled with the riboswitch, which was originally ordered from Genewiz. The constitutive promoter [J23100] has ~99% promoter efficiency and was tested using GFP assays in both induced and uninduced promoters.








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