Coding

Part:BBa_K3944021:Design

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


TEF1-TetA-ScPGK1


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 760
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1418
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1331
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 161


Design Notes

The chimeric protein must be assembled scarless, but the promoter/terminator regions may be assembled with biobrick-compatible scars. Promoter should be constitutive since this is a sensing protein required for tetracycline induction.


Source

Assembled from parts provided by systems biology department at Chalmers technical highschool. Parts originate from bacteria and yeast.

References

1: Gemma Bellí, Eloi Garí, Lidia Piedrafita, Martí Aldea, Enrique Herrero, An activator/repressor dual system allows tight tetracycline-regulated gene expression in budding yeast, Nucleic Acids Research, Volume 26, Issue 4, 1 February 1998, Pages 942–947, https://doi.org/10.1093/nar/26.4.942
2: Hirai H, Tani T, Kikyo N. Structure and functions of powerful transactivators: VP16, MyoD and FoxA. Int J Dev Biol. 2010;54(11-12):1589-1596. doi:10.1387/ijdb.103194hh
3: Traven A, Jelicic B, Sopta M. Yeast Gal4: a transcriptional paradigm revisited. EMBO Rep. 2006;7(5):496-499. doi:10.1038/sj.embor.7400679
4: Irina L. Rempel, Petra Popken, Ali Ghavami, Ankur Mishra, Rizqiya A. Hapsari, Anouk H.G. Wolters, Annemiek C. Veldsink, Marindy Klaassens, Anne C. Meinema, Bert Poolman, Ben N.G. Giepmans, Patrick R. Onck, Anton Steen, Liesbeth M. Veenhoff, Flexible and Extended Linker Domains Support Efficient Targeting of Heh2 to the Inner Nuclear Membrane, Structure, Volume 28, Issue 2, 2020, Pages 185-195.e5, ISSN 0969-2126, https://doi.org/10.1016/j.str.2019.11.003.
5: Partow, S., Siewers, V., Bjørn, S., Nielsen, J. and Maury, J. (2010), Characterization of different promoters for designing a new expression vector in Saccharomyces cerevisiae. Yeast, 27: 955-964. https://doi.org/10.1002/yea.1806