Composite

Part:BBa_K4890012:Design

Designed by: Duoqing LIN   Group: iGEM23_Shanghai-SDG   (2023-08-14)


MRE-Hsp70


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 204
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 204
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 204
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 204
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We used pUAST vector system to construct pMRE-Hid plasmid. pUAST vector contains UAS and Hsp70. We replaced UAS with MRE in the pUAST vector and inserted Hid gene in the modified pUAST vector downstream of MRE and Hsp70 by enzyme cutting and linking. The plasmid was then transfected into Drosophila S2 cells. We also inserted MTF-1 gene into the cells. MTF-1 protein activated by heavy metals in the environment can bind to MRE and trigger the expression of Hid gene.


Source

MRE sequence is based on BBa_K4890002. Hsp70 sequence is based on BBa_K4890004.


References