Translational_Unit

Part:BBa_K2387078

Designed by: Bart Scholten   Group: iGEM17_Wageningen_UR   (2017-10-27)


TEV cleavable CpxA-eYFP[1-154]-nZIP and CpxA-eYFP[155-238]-cZIP+ araC/pBAD promoter

In order to visualize Cpx pathway activation, transmembrane signal transducing protein CpxA is fused to respectively eYFP(1-154) and N-Leucine Zipper [1] ,as well as eYFP(155-238) and C-Leucine Zipper [2]. CpxA and eYFP[1-154] /eYFP[155-238] are fused with a TEV cleavable peptide. When cotransformed with CpxR-TEV protease fusion activation of the Cpx pathway is visualized in combination with CpxA-eYFP[1-154]-N-Leucine Zipper fusion due to eYFP reassembly.

Usage and Biology

This BioBrick was used following this protocol.

Figure 1: CpxA-CpxR protein interaction visualized with different L-arabinose concentrations over time. Upon Cpx activation, eYFP-termini are cleaved off of CpxA by TEV protease and reassemble in the cytoplasm using leucine zipper's natural affinity for each other.

More information can be found here


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1144
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 979
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961


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Parameters
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