Part:BBa_K819005

Constitutive LuxBrick Generator

Summary

Design

Characterization

Reference

Summary

This part, which is a fusion protein, serves as an ultra sensitive photoreceptor (which can sense light as weak as moonlight) and will induce a light-dependent repression of genes under specific promoters like RecA408 promoter and SulA408 promoter.

For the photosensor domain of this fusion protein, we chose VVD protein, the smallest photosensor protein known, which forms a rapidly exchanging dimer when excited by blue light. For the physiological function domain, we chose LexA protein, a bacteria transcription inhibitor, whose DNA binding domain rigorously requires help of the dimerization domain for its DNA binding activity. We fused these two separate domains together to form a fusion protein, which we termed our "Luminesensor". Both DNA binding domain and dimerization domain of this luminesensor was mutated at specific sites for better bio-orthogonality and higher on-off ratio.(to see the design page for details, please click here)

We then characterized this luminesensor in different ways and all the results showed that this luminesensor could response to light quite well. (to see the characterization page for details, please click here)

To learn more details about this luminesensor and the related project by Peking 2012 iGEM team, please visit our [http://2012.igem.org/Team:Peking wiki]

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 3014
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 2012
  Illegal XhoI site found at 2842
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 4401
  Illegal BsaI.rc site found at 1410
  Illegal SapI.rc site found at 4726