Plasmid

Part:BBa_K3482026

Designed by: Thierry Marti   Group: iGEM20_UNILausanne   (2020-10-23)


pLPT41 sponge plasmid(Potvin-Trottier, 2016)

This plasmid acts a molecular titration sponge for TetR. It is intended to be used with a repressilator plasmid. It stabilizes the oscillations of the transcription factors of the repressilator.

Usage and Biology

The repressilator is a synthetic regulatory network of at least three genes in a plasmid that repress each other in a feedback loop. These genes are cyclically expressed one after the other in a loop at regular intervals over time. This property of cyclic expression of each gene allows us to produce our anti-cancer drug regularly for cancer treatment. For our system to work efficiently, the sponge plasmid is applied together with the repressilator to smooth out the observed oscillations by buffering excess proteins produced by the main repressilator.

All of the plasmids presented here were obtained and ordered from Addgene. They were all made available by the Professor Johan Paulson. All the plasmids come form this article Potvin-Trottier, L., Lord, N., Vinnicombe, G. et al. Synchronous long-term oscillations in a synthetic gene circuit. Nature 538, 514–517 (2016). https://doi.org/10.1038/nature19841.

Characterization

All the data concerning the characterization of that part can be found in the composite part : BBa_K3482023 and BBa_K3482025 .

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 87
    Illegal EcoRI site found at 172
    Illegal SpeI site found at 1133
    Illegal PstI site found at 121
    Illegal PstI site found at 178
    Illegal PstI site found at 1882
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 87
    Illegal EcoRI site found at 172
    Illegal SpeI site found at 1133
    Illegal PstI site found at 121
    Illegal PstI site found at 178
    Illegal PstI site found at 1882
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 87
    Illegal EcoRI site found at 172
    Illegal BglII site found at 2095
    Illegal BamHI site found at 190
    Illegal XhoI site found at 7
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 87
    Illegal EcoRI site found at 172
    Illegal SpeI site found at 1133
    Illegal PstI site found at 121
    Illegal PstI site found at 178
    Illegal PstI site found at 1882
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 87
    Illegal EcoRI site found at 172
    Illegal SpeI site found at 1133
    Illegal PstI site found at 121
    Illegal PstI site found at 178
    Illegal PstI site found at 1882
    Illegal NgoMIV site found at 1431
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 1371
    Illegal SapI site found at 1581


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