Difference between revisions of "Part:BBa K4060106:Design"

 
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===Design Notes===
 
===Design Notes===
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The construction of this part is according to igem assembly protocol.
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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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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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    <img src="https://2021.igem.org/wiki/images/d/d4/T--NYCU-Taipei--S3.png" style="padding-bottom:2%;width:100%">
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    <span class="caption">Fig1. Design logic of our composite parts.</span>
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===Source===
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The parts are from 2019 igem kits.
  
 
===References===
 
===References===
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1. 2020 Fudan iGEM team, https://2020.igem.org/Team:Fudan/Design/kill_switch<p></p>

Latest revision as of 14:26, 7 December 2021


Pre B3 gene


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]


Design Notes

The construction of this part is according to igem assembly protocol. 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.

Fig1. Design logic of our composite parts.

Source

The parts are from 2019 igem kits.

References

1. 2020 Fudan iGEM team, https://2020.igem.org/Team:Fudan/Design/kill_switch