Difference between revisions of "Part:BBa K4604037:Design"

 
 
(3 intermediate revisions by the same user not shown)
Line 1: Line 1:
 +
__NOTOC__
 +
<partinfo>BBa_K4604037 short</partinfo>
  
 +
<partinfo>BBa_K4604037 SequenceAndFeatures</partinfo>
 +
 +
 +
===Design Notes===
 +
The methionine start codon was shifted to fit a HA-tag in the reading frame. 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 <i>E. coli</i> K12 MG1655 genome, synthesised by IDT. Sequence taken from National Library of Medicine (gene ID:947245).
 +
 +
===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

Latest revision as of 01:10, 11 October 2023

MazE


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 89
  • 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 HA-tag in the reading frame. 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:947245).

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