Difference between revisions of "Part:BBa K4604011:Design"

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===Design Notes===
 
===Design Notes===
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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.
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The methionine start 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===
 
===Source===
  
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Occurs naturally in E. coli K12 MG1655 genome, synthesised by IDT. Sequence taken from National Library of Medicine.  
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Occurs naturally in <i>E. coli</i> K12 MG1655 genome, synthesised by IDT. Sequence taken from National Library of Medicine (gene ID:947252).  
  
 
===References===
 
===References===
 
[1] The Regulation of the Escherichia coli mazEF Promoter Involves an Unusual Alternating Palindrome *
 
[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
 
Marianovsky, Irina et al. Journal of Biological Chemistry, Volume 276, Issue 8, 5975 - 5984

Latest revision as of 01:10, 11 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

The methionine start 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 (gene ID:947252).

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