Composite

Part:BBa_K4387987:Design

Designed by: Nathalie Weibel   Group: iGEM22_UZurich   (2022-09-29)


Hemolysin A secretion system for E. coli


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1160
    Illegal AgeI site found at 1330
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We decided to use the constitutive promoter J23100 to ensure proper channel formation and faster secretion of our therapeutics, thanks to the already existing secretion machinery in our bacteria. To decrease a bit the burden on our bacteria, we integrated the secretion system in a medium copy number backbone (p15A) and order it from Twist Bioscience.


Source

Most parts were taken from the paper by Ruano-Gallego et al. [1]. More information can be found on the specific part sites.


References

  • [1] Ruano-Gallego, D., Fraile, S., Gutierrez, C. et al. Screening and purification of nanobodies from E. coli culture supernatants using the hemolysin secretion system. Microb Cell Fact 18, 47 (2019). https://doi.org/10.1186/s12934-019-1094-0