Difference between revisions of "Part:BBa K3408009"
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<partinfo>BBa_K3408009 short</partinfo>
 
<partinfo>BBa_K3408009 short</partinfo>
  
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This composite part consists of promoter P<sub>liaG</sub> and P<sub>grac</sub>, lacI gene, C&#8544; gene, GFP gene and some essential RBS and terminators. Promoter P<sub>liaG</sub> starts transcription process and its transcription production is LacI, which is regulated by IPTG. When bacteria is exposed to medium which is rich in IPTG, bacteria can intake IPTG. IPTG is combined with LacI, then LacI cannot be bound to promoter Pgrac which is specially suppressed by LacI. Therefore, C&#8544; repressor protein cannot generate as downstream cI gene cannot transcribe regularly.
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We used constitutive promoter P<sub>liaG</sub> to transcribe trigger RNA all the time. And we also used P<sub>CⅠ</sub> as a constitutive promoter to transcribe switch RNA. If our Toehold switch could work normally, GFP could be translated and we could see green bacteria in our culture medium.
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As a result, P<sub>C&#8544;</sub> recovers its activity and triggers the downstream transcription of GFP. By checking the expression of GFP, we could verify the feasibility of IPTG induction system, thus guaranteeing the successful culture of our engineered <i>Bacillus subtilis</i>.  
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Plasmid profile
 
Plasmid profile
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https://2020.igem.org/wiki/images/5/58/T--NAU-CHINA--composite-parts6.1.png
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https://2020.igem.org/wiki/images/e/ea/T--NAU-CHINA--composite-parts4.1.png
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
 
===Usage and Biology===
 
===Usage and Biology===

Revision as of 14:02, 16 October 2020

PliaG-trigger RNA-B0015-PCⅠ-switch RNA-GFP-B0015

We used constitutive promoter PliaG to transcribe trigger RNA all the time. And we also used PCⅠ as a constitutive promoter to transcribe switch RNA. If our Toehold switch could work normally, GFP could be translated and we could see green bacteria in our culture medium.

Plasmid profile T--NAU-CHINA--composite-parts4.1.png 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 239
    Illegal BglII site found at 461
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1190