Part:BBa_K2623011

Phosphorylation pathsway builder of Kai

Summary

The sequence encoding for the gene is preceded by a promoter BBa_J23100, an RBS BBa_B0034, BBa_K2623004, BBa_K2623005, BBa_K2623006 and followed by a double terminator BBa_B0015 in pSB1C3 to pSB1C3 to convert the periodic oscillation of KaiC phosphorylation into the downstream transcriptional oscillation using the signal pathways of CikA, SasA and RpaA.

Fig.1 Simplified model of the central circadian oscillator and output activities.

Indentification

In order to verify the effectiveness of our Kai system, we verified the Escherichia coli that was transferred into the Kai loop under the fluorescence microscope after synchronization.
Since the Kai system is designed for periodic output of signals in Escherichia coli, we evaluateD the changes of fluorescent signals over time. Here are the curves we made. More details can be found in http://2018.igem.org/Team:XMU-China/Measurement

Fig.3 The bioluminescence at different times in GroupA. The negative value is cause by the value of the control group.	Fig.4 The bioluminescence at different times in GroupB. The negative value is cause by the value of the control group.

Fig.5 The fitting curve of the oscillator in GroupA	Fig.6 The fitting curve of the oscillator in GroupB

More details can be seen in http://2018.igem.org/Team:XMU-China/Results

Sequence and Features

Assembly Compatibility:

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

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

  Illegal BglII site found at 188
  Illegal BglII site found at 3666
  Illegal BglII site found at 3735
  Illegal BamHI site found at 2248
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]