Part:BBa_K4767013

Pars-RBS-arsR-RBS-luxR(△2-162)-TT-PluxⅠ-RBS-luxR(△2-162)-RBS-gfp -TT

Uses a factor P_ars(BBa_K4767001) ,strong RBS(BBa_J34801), DNA binding transcriptional repressor arsR(BBa_J15101), luxR(△2-162)(BBa_K4767000) TT(BBa_B0015) ,P_luxⅠ(BBa_R0062) and gfp(BBa_E0040).This composite part is a BioBrick.

Usage and Biology

In order to construct a positive feedback circuit which does not require 3OC6HSL produced by LuxI, we engineered LuxR by deleting 2-262 amino acids in the N-terminal domain (AHL binding domain) and reserving a C-terminal domain with the function of activating transcription, obtaining a resulting regulator LuxR(△2-162). LuxR(△2-162) can active the gene transcription driven by the lux promoter in the absence of AHL. To construct the amplifier, we cloned gfp and LuxR(Δ2-162) behind the lux promoter. In this design, LuxR(Δ2-162) functions in a positive feedback loop as it can bind to the P_luxⅠ promoter and activate its own transcription.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 270
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 929
  Illegal BsaI.rc site found at 1952
  

References

Xiaoqiang Jia, Bu Rongrong, Zhao Tingting, et al. Sensitive and Specific Whole-Cell Biosensor for Arsenic Detection[J]. Applied and environmental microbiology, 2019, 85(11): 1.

Nistala, G.J., et al., A modular positive feedback-based gene amplifier. J Biol Eng, 2010. 4: p. 4.