Difference between revisions of "Part:BBa K4137002:Design"

 
(→‎Design Notes)
Line 7: Line 7:
  
 
===Design Notes===
 
===Design Notes===
−
The 6xHis tag was added to the mleR coding sequence during part design, as we wanted to characterize the toxin-antitoxin system and thus had to derive a parameter for its quantity; we thus needed to perform protein quantification, and believed adding a 6xHis tag to be the most straightforward method of achieving this end.
+
The 6xHis tag was added to the mleR coding sequence during part design, as we wanted to characterize the toxin-antitoxin system and thus had to derive a parameter for its quantity; we, therefore, needed to perform protein quantification and believed adding a 6xHis tag to be the most straightforward method of achieving this end.
−
The sequence of mleR and 6xHis was then reversed, as we would go on to use it in the composite part BBa_K4130, which uses a T7 promoter to overexpress mleR; the T7 promoter is optimal when reversed, so we adjusted this part's direction in response to this fact.
+
The sequence of mleR and 6xHis was then reversed, as we would use it in the composite part BBa_K4130, which uses a T7 promoter to overexpress mleR; the T7 promoter is optimal when reversed, so we adjusted this part's direction in response to this fact.
−
 
+
  
 +
Unfortunately, the sequence features view tool does not reflect the fact that this part is reversed; attached below is a benchling screenshot of how the sequence features should look.
  
 
===Source===
 
===Source===

Revision as of 04:14, 9 August 2022


Malate-binding Transcriptional Activator + 6xHis


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 18
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The 6xHis tag was added to the mleR coding sequence during part design, as we wanted to characterize the toxin-antitoxin system and thus had to derive a parameter for its quantity; we, therefore, needed to perform protein quantification and believed adding a 6xHis tag to be the most straightforward method of achieving this end. The sequence of mleR and 6xHis was then reversed, as we would use it in the composite part BBa_K4130, which uses a T7 promoter to overexpress mleR; the T7 promoter is optimal when reversed, so we adjusted this part's direction in response to this fact.

Unfortunately, the sequence features view tool does not reflect the fact that this part is reversed; attached below is a benchling screenshot of how the sequence features should look.

Source

https://journals.asm.org/doi/10.1128/jb.171.6.3108-3114.1989

References