Composite
Pre B3

Part:BBa_K4060106:Design

Designed by: YU CHUAN LEE   Group: iGEM21_NYCU-Taipei   (2021-09-30)
Revision as of 03:39, 22 October 2021 by JasonCTS (Talk | contribs)


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]
    Unknown
  • 23
    INCOMPATIBLE WITH RFC[23]
    Unknown
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 711
    Illegal AgeI site found at 823
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

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.


Source

References