Part:BBa_K2387029
CpxR-mVenus[1-157] and CpxR-mVenus[158-238] + araC/pBAD promoter
The [http://parts.igem.com/Part:BBa_K2387038 N-terminus of mVenus] and the [http://parts.igem.com/Part:BBa_K2387039 C-terminus of mVenus] are fused to Cpx response regulator [http://parts.igem.com/Part:BBa_K2387002 CpxR] in order to visualize the activation of the Cpx pathway. Upon activation of the Cpx pathway, CpxR gets phosphorylated by E. coli endogenous CpxA after which it can homodimerize. This protein-protein interaction can be visualized using BiFC, hence the fusion of mVenus[1-157] and mVenus[158-238]. The fusion is put under the control of the L-arabinose inducible araC/pBAD promoter. Strong RBS BBa_B0034 is used to regulate transcription.
CpxR-mVenus[1-157] and CpxR-mVenus[158-238] fusions were linked together using a [http://parts.igem.com/Part:BBa_K1486004 Flexible Linker] consisting of two times the amino acids GGGGS.
This part is used to visualize the activation of the Cpx pathway via Bimolecular Fluorescence Complementation by using the CpxR-CpxR interaction.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1205
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1144
Illegal XhoI site found at 1283
Illegal XhoI site found at 2498 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 979
Illegal AgeI site found at 1694
Illegal AgeI site found at 2909 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 961
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