Generator

Part:BBa_K1961004

Designed by: Yi-Ting Lin   Group: iGEM16_HSiTAIWAN   (2016-10-03)


PbrR generator expressed by GFP under autoregulation of PTet

The Pb2+ biosensor involves (1) generation of PbrR promoter with a binding site for lead ion, thus forming Pb2+-PbrR dimer and (2) GFP expression when Pb2+-PbrR dimer binds onto pbr promoter. Because the PbrR and Pb2+ - PbrR dimer compete to bind onto pbr promoters and result in poor GFP performance, we designed an autoregulated PbrR generator (BBa_K1961004) to control the amount of PbrR.

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This composite part consists of the TetR and PbrR generators, both controlled by PTet, which has TetR binding sites. TetRs bind onto PTet and repress the gene expression of TetR, PbrR and GFP. We used tetracycline to induce the gene expression of PbrR and GFP.

We tested the survival of E.coli in tetracycline(TC). E. coli can survival in TC of 1000 ng/ml. In the following figure, it shows that GFP expression of autoregulation is slower than that with constitutive promoter(BBa_K1961005)

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Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2040


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