Difference between revisions of "Part:BBa K4604011:Design"
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===Design Notes===
 
===Design Notes===
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First methionine codon was removed 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. We therefore decided to use a different promoter than the native one to prevent interference by the toxin-antitoxin complex.  
+
First methionine codon was removed 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.  
  
  
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===References===
 
===References===
 +
[1] The Regulation of the Escherichia coli mazEF Promoter Involves an Unusual Alternating Palindrome *
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Marianovsky, Irina et al. Journal of Biological Chemistry, Volume 276, Issue 8, 5975 - 5984

Revision as of 16:22, 7 October 2023


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 removed 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 Medizine.

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