Difference between revisions of "Part:BBa K4137002:Design"
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However, for registry documentation purposes, the sequenced attached here is in the forward direction; attached below is a benchling diagram of the sequence as it is used in our project experimentation
 
However, for registry documentation purposes, the sequenced attached here is in the forward direction; attached below is a benchling diagram of the sequence as it is used in our project experimentation
  
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[[Image:mleR_benchling.png|750px|thumb|center|Figure 1. Benchling Linear Map View of the BBa_K4137002 Basic Part.]]
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[[File:ccda-his.png|800px|thumb|center|Fig.1 ccdA with C-Terminal 6x His-Tag.]]
  
 
===Source===
 
===Source===

Revision as of 03:08, 9 October 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 needed to purify mleR and then quantify its production to characterize the toxin-antitoxin system through the derivation of kinetic constants.

The sequence of mleR and 6xHis was then reversed alongside the entirety of BBa_K4137008 such that the transcription of CcdA would not interfere with mleR, and vice versa; this was taken into our design as our constructs only have a single terminator. This sequence is codon optimized for E. coli.

However, for registry documentation purposes, the sequenced attached here is in the forward direction; attached below is a benchling diagram of the sequence as it is used in our project experimentation

Fig.1 ccdA with C-Terminal 6x His-Tag.

Source

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

References

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