Part:BBa_K2197300:Design

Expression of GFP under the control of a uric acid concentration-sensitive promoter device

Assembly Compatibility:

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

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  Illegal NheI site found at 1098
  Illegal NheI site found at 1121
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal XhoI site found at 555
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 291
  Illegal BsaI.rc site found at 1814
  Illegal SapI.rc site found at 255
  

Design Notes

Part BBa_K2197300 can be divided into two sessions, the promoter and the GFP expression. The promoter is designed to be sensitive to the concentration of uric acid. This promoter control the expression of GFP that is downstream the promoter. The promoter session consists of a constitutive promoter J23100, a RBS B0034, a strong repressor KRAB-HucR, a double terminator B0015. HucR is itself a repressor. Its repressing ability is enhanced by KRAB. The resulting repressor is a chimeric mammalian urate-dependent transsilencer (mUTS). hucO is an operative site for mUTS to bind. When mUTS is binded to hucO, the expression of downstream gene is restricted according concentration of substrates. The presence of uric acid limits the binding of mUTS to hucO. The limitation varies as the concentration of uric acid. Downstream of the promoter session is a constitutive promoter J23106, a RBS B0034, a GFP gene E0040 and a terminator B1002. As a result mUTS binds to hucO and GFP is not expressed when uric acid is absent or at very low concentration. Alternatively, the complex detaches from hucO and GFP is expressed according to the concentration of uric acid. The promoter control the expression of GFP. Engineered e.coli encodes part BBa_K2197300.

Source

Deinococcus radiodurans R1

References

http://www.nature.com/nbt/journal/v28/n4/full/nbt.1617.html