Part:BBa_K2387010

CpxA-eYFPn[1-154] fusion + araC/pBAD promoter

The [http://parts.igem.com/Part:BBa_K2387005 N-terminus of eYFP] is fused to transmembrane signal transducer [http://parts.igem.com/Part:BBa_K2387002 CpxA] in order to visualize the activation of the Cpx pathway. The fusion is put under the control of the L-arabinose inducible araC/pBAD promoter. Strong RBS BBa_B0034 is used to regulate transcription.

CpxA and eYFPn were linked together using a [http://parts.igem.com/Part:BBa_K1486004 Flexible Linker] consisting of two times the amino acids GGGGS.

It can be used in combination with the fusion [http://parts.igem.com/Part:BBa_K2387011 CpxR-eYFPc[155-238]] to visualize the activation of the Cpx pathway via Bimolecular Fluorescence Complementation

Usage and Biology

This BioBrick (in combination with BBa_K2387011) was used following this protocol.

Figure 1: CpxA-CpxR protein interaction visualized with different L-arabinose concentrations over time.

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