Device

Part:BBa_K5127014:Design

Designed by: Chujing Wu   Group: iGEM24_BNDS-China   (2024-10-02)


Butyrate sensor with NOT gate (pHpdH-ci-HpdR)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1237
    Illegal NotI site found at 408
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 392
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 2386
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

To integrate the butyrate biosensor into our ultimate project goal, we designed the CI gene into the pHpdH sensing system plasmid. In this system, when butyrate levels are low, the output of GFP will be higher (Figure 1). This design enables us to monitor and respond to butyrate concentration effectively within our system. We contructed this plasmid by using Golden Gate Assembly (Figure 2).


Source

When butyrate is present, it binds to HpdR, changing its conformation so it could bind and activate the pHpdH promoter, resulting in expression of downstream genes, in which GFP downstream of the plam promoter will be expressed if butyrate concentration is relatively low. This is an inverse logic of the part pHpdR (BBa_K2560304).


References

N/A