Regulatory

Part:BBa_K4129115:Design

Designed by: Magnus Haahr   Group: iGEM22_DTU-Denmark   (2022-10-09)


Minimal gpdA promoter fused with six LexO bindings sites (6xLexO-Pmin)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 11
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 11
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 11
  • 1000
    COMPATIBLE WITH RFC[1000]

Design Notes

This synthetic promoter had too high complexity to be ordered as a gene block. It was assembled from plasmids.

Source

This part was assembled using USER cloning. The minimal gpdA promoter was amplifed from an in-house plasmid carring the TetOn system. The six lexO binding sites, 6xLexO, was amplifed from a in-house plasmid granted to us from Michael Krogh Jensen and Marcus Deichmann from The Novo Nordisk Foundation Center for Biosustainability.

References

Radman M. SOS repair hypothesis: phenomenology of an inducible DNA repair which is accompanied by mutagenesis. Basic Life Sci. 1975;5A:355-67. doi: 10.1007/978-1-4684-2895-7_48. PMID: 1103845.

Rantasalo A, Landowski CP, Kuivanen J, Korppoo A, Reuter L, Koivistoinen O, Valkonen M, Penttilä M, Jäntti J, Mojzita D. A universal gene expression system for fungi. Nucleic Acids Res. 2018 Oct 12;46(18):e111. doi: 10.1093/nar/gky558. PMID: 29924368; PMCID: PMC6182139.

Wanka F, Cairns T, Boecker S, Berens C, Happel A, Zheng X, Sun J, Krappmann S, Meyer V. Tet-on, or Tet-off, that is the question: Advanced conditional gene expression in Aspergillus. Fungal Genet Biol. 2016 Apr;89:72-83. doi: 10.1016/j.fgb.2015.11.003. Epub 2015 Nov 10. PMID: 26555930.