Coding

Part:BBa_K3944035:Design

Designed by: Olof Dahlman   Group: iGEM21_Chalmers-Gothenburg   (2021-09-30)


TEF1p-Estra-ScADH1t


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 580
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 161


Design Notes

Estradiol-binding protein must be assembled scarless as it is a fusion protein, but promoter and terminator may be swapped and assembled at will. Promoter and terminator are native to yeast, while the fusion protein has been optimised for use in S.Cerevisiae.


Source

Parts provided by systems biology at Chalmers technical highschool and assembled using Gibson assembly methods.

References

1: R. Scott McIsaac, Patrick A. Gibney, Sunil S. Chandran, Kirsten R. Benjamin, David Botstein, Synthetic biology tools for programming gene expression without nutritional perturbations in Saccharomyces cerevisiae, Nucleic Acids Research, Volume 42, Issue 6, 1 April 2014, Page e48, https://doi.org/10.1093/nar/gkt1402
2: Fast-acting and nearly gratuitous induction of gene expression and protein depletion in Saccharomyces cerevisiae R. Scott McIsaac, Sanford J. Silverman, Megan N. McClean, Patrick A. Gibney, Joanna Macinskas, Mark J. Hickman, Allegra A. Petti, and David Botstein Molecular Biology of the Cell 2011 22:22, 4447-4459
3: Ottoz DS, Rudolf F, Stelling J. Inducible, tightly regulated and growth condition-independent transcription factor in Saccharomyces cerevisiae. Nucleic Acids Res. 2014;42(17):e130. doi: 10.1093/nar/gku616. Epub 2014 Jul 17. PMID: 25034689; PMCID: PMC4176152.