Difference between revisions of "Part:BBa K4767015"
 
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<partinfo>BBa_K4767015 short</partinfo>
 
<partinfo>BBa_K4767015 short</partinfo>
  
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Uses a factor PluxⅠ(BBa_R0062) ,strong RBS(BBa_J34801),  <i>luxR</i>(△2-162)(BBa_K4767000), and TT(BBa_B015). This composite part is a BioBrick.
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Uses a factor P<i><sub>luxⅠ</sub></i>(BBa_R0062) ,strong RBS(BBa_J34801),  <i>luxR</i>(△2-162)(BBa_K4767000), and TT(BBa_B0015). This composite part is a BioBrick.
  
  
 
===Usage and Biology===
 
===Usage and Biology===
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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 <i>lux</i> promoter in the absence of AHL. To construct the amplifier, we cloned LuxR(Δ2-162) behind the <i>lux</i> promoter. In this design, LuxR(Δ2-162) functions in a positive feedback loop as it can bind to the PluxI promoter and activate its own transcription.  
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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 <i>lux</i> promoter in the absence of AHL. To construct the amplifier, we cloned LuxR(Δ2-162) behind the <i>lux</i> promoter. In this design, LuxR(Δ2-162) functions in a positive feedback loop as it can bind to the P<i><sub>luxⅠ</sub></i> promoter and activate its own transcription.  
  
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<center>[[File:BBa_K4767015.jpg]]</center>
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<center>https://static.igem.wiki/teams/4767/wiki/part/img-1179.png</center>
  
 
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Latest revision as of 08:35, 8 October 2023


PluxⅠ-RBS-luxR(△2-162)-TT

Uses a factor PluxⅠ(BBa_R0062) ,strong RBS(BBa_J34801), luxR(△2-162)(BBa_K4767000), and TT(BBa_B0015). 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 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 PluxⅠ promoter and activate its own transcription.

img-1179.png

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

References

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