Difference between revisions of "Part:BBa K4060106"
 
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<partinfo>BBa_K4060106 short</partinfo>
 
<partinfo>BBa_K4060106 short</partinfo>
  
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In our Design 1, antitoxin MazE was chosen as the core effector component, with pBad promoter and RNA thermometer acting as sensor components, and the TetR gene and TetR repressible promoter working together as an inverting logic gate. If all goes as expected, MazE will express under 37°C if the medium is free of L-arabinose, but the expression level will drop significantly either with the presence of L-Arabinose or when the ambient temperature is low. To evaluate the behavior of our system, we created three constructs:  
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a kill switch strategy. In our Design 1, antitoxin MazE was chosen as the core effector component, with pBad promoter and RNA thermometer acting as sensor components, and the TetR gene and TetR repressible promoter working together as an inverting logic gate. If all goes as expected, MazE will express under 37°C if the medium is free of L-arabinose, but the expression level will drop significantly either with the presence of L-Arabinose or when the ambient temperature is low. To evaluate the behavior of our system, we created three constructs:  
 
Firstly, we put all parts together as our plan in Constriuct 1. In Construct 2 , we would like to visualize how MazE will express under different L-Arabinose concentration and ambient temperature for quick check and further assays, so we replaced MazE with RFP. By the same token, we replaced TetR with RFP in Construct 3 to evaluate the expression level of TetR under different concentrations of L-Arabinose in case we needed to troubleshoot the system.
 
Firstly, we put all parts together as our plan in Constriuct 1. In Construct 2 , we would like to visualize how MazE will express under different L-Arabinose concentration and ambient temperature for quick check and further assays, so we replaced MazE with RFP. By the same token, we replaced TetR with RFP in Construct 3 to evaluate the expression level of TetR under different concentrations of L-Arabinose in case we needed to troubleshoot the system.
  
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<!-- Add more about the biology of this part here
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<!-- Add more about the biology of this part here-->
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In our Design 1, antitoxin MazE was chosen as the core effector component, with pBad promoter and RNA thermometer acting as sensor components, and the TetR gene and TetR repressible promoter working together as an inverting logic gate. If all goes as expected, MazE will express under 37°C if the medium is free of L-arabinose, but the expression level will drop significantly either with the presence of L-Arabinose or when the ambient temperature is low. To evaluate the behavior of our system, we created three constructs:  
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===Usage and Biology===
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Firstly, we put all parts together as our plan in Constriuct 1. In Construct 2 , we would like to visualize how MazE will express under different L-Arabinose concentration and ambient temperature for quick check and further assays, so we replaced MazE with RFP. By the same token, we replaced TetR with RFP in Construct 3 to evaluate the expression level of TetR under different concentrations of L-Arabinose in case we needed to troubleshoot the system.
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The RFP expression of Construct 3
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We used RFP(E1010) to monitor the expression of TetR, so that we can figure out how L-arabinose will influence the expression of TetR in our Design 1.
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<html><body>
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<img src=" https://2021.igem.org/wiki/images/f/fb/T--NYCU-Taipei--kilksw_result_1.png" height="475px" width="750px"/>
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</body></html>
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<html><body>
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<img src="https://2021.igem.org/wiki/images/3/34/T--NYCU-Taipei--kilksw_result_2.png" height="475px" width="750px"/>
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</body></html>
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K4060106 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K4060106 SequenceAndFeatures</partinfo>

Latest revision as of 14:49, 7 December 2021


Pre B3 gene

a kill switch strategy. In our Design 1, antitoxin MazE was chosen as the core effector component, with pBad promoter and RNA thermometer acting as sensor components, and the TetR gene and TetR repressible promoter working together as an inverting logic gate. If all goes as expected, MazE will express under 37°C if the medium is free of L-arabinose, but the expression level will drop significantly either with the presence of L-Arabinose or when the ambient temperature is low. To evaluate the behavior of our system, we created three constructs: Firstly, we put all parts together as our plan in Constriuct 1. In Construct 2 , we would like to visualize how MazE will express under different L-Arabinose concentration and ambient temperature for quick check and further assays, so we replaced MazE with RFP. By the same token, we replaced TetR with RFP in Construct 3 to evaluate the expression level of TetR under different concentrations of L-Arabinose in case we needed to troubleshoot the system.

Usage and Biology

The RFP expression of Construct 3 We used RFP(E1010) to monitor the expression of TetR, so that we can figure out how L-arabinose will influence the expression of TetR in our Design 1.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 125
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 65
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 711
    Illegal AgeI site found at 823
  • 1000
    COMPATIBLE WITH RFC[1000]