Coding

Part:BBa_K4604011:Design

Designed by: Hannah Swientek   Group: iGEM23_Freiburg   (2023-09-30)
Revision as of 12:32, 10 October 2023 by NikitaEdel (Talk | contribs) (→‎Design Notes)


MazF toxin


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

First methionine codon was shifted to fit a FLAG-tag in the reading frame. Stop codon was also removed to guarantee translation of reporter gene afterwards. It’s important to note that the toxin-antitoxin system MazE/F is already present and constantly expressed in the strain we use, therefore a baseline expression of the native system is given. The endogenous MazE/F system regulates its own gene expression [1]. We therefore decided to use a different promoter than the native one to prevent interference by the toxin-antitoxin complex.

Source

Occurs naturally in E. coli K12 MG1655 genome, synthesised by IDT. Sequence taken from National Library of Medicine.

References

[1] The Regulation of the Escherichia coli mazEF Promoter Involves an Unusual Alternating Palindrome * Marianovsky, Irina et al. Journal of Biological Chemistry, Volume 276, Issue 8, 5975 - 5984