Composite

Part:BBa_K2486019

Designed by: Cauă Westmann, Gabriel Lencioni Lovate   Group: iGEM17_USP-Brazil   (2017-10-24)


PlldR_tetR/LldR based lactate biosensor

A lactate biosensor composed by parts BBa_K2486007 and BBa_K24860010. The part BBa_K2486007 works as a LldR generator, one of the repressor proteins capable of blocking gene expression of our PlldR_tetR promoter (the other protein is TetR). The other componet of our biosensor is the part BBa_K24860010 a reporter module composed of our hybrid promoter (PlldR_tetR promoter) and a GFP::lva.

Characterization

T--USP-Brazil--Detecttion_Iron_Lactate_concept_%285%29.PNG

The data of the validation of this part in the presence of LldR and TetR is described in parts BBa_K2486019 (LldR-based lactate biosensor) and BBa_K2486020 (TetR-based tetracycline biosensor) and here.

PlldR_tetR promoter activity in the presence of LldR regulator

800px-T--USP-Brazil--Detecttion_Iron_Lactate_lldR%281%29.png

PlldR_tetR promoter activity in the presence of TetR regulator

T--USP-Brazil--Detecttion_Iron_Lactate_TetR%282%29.png

Conclusion

The presence of LlldR and lactate affects the circuit as expected, repressing it in the absence of lactate. The presence of TetR and tetracycline worked as expected too, sugesting that the part could work with both regulators.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 874
    Illegal NheI site found at 897
    Illegal NheI site found at 1033
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 284
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1743


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Categories
Parameters
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