Difference between revisions of "Part:BBa K3866018"
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===References===
 
===References===
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*Choi, Y. J., Morel, L., Le François, T., Bourque, D., Bourget, L., Groleau, D., Massie, B., & Míguez, C. B. (2010). Novel, versatile, and tightly regulated expression system for Escherichia coli strains. Applied and environmental microbiology, 76(15), 5058–5066. https://doi.org/10.1128/AEM.00413-10

Latest revision as of 10:53, 29 September 2021


J23115-RBS-CymR-double terminator

Fig.2:The Level A of CymR with the J23115 Anderson promoter and the double terminator in an alpha1R vector.

Usage and Biology

The Cym repressor is produced with the aid of the strong constitutive promoter J23115 BBa_K3505012 and the double terminator BBa_K3505017. The Cym Repressor binds to the cumate operator CuO that is downstream the partial T5 promoter and causes the production of the toxin mazF and by extension the death of the bacteria.


Design Notes

The coding sequence was domesticated . We removed BsmBI ,BsaI , BtgZI, BpiI sites in order to be compatible with GoldenBraid and MoClo. The sequence is cloned in alpha1R vector BBa_K3505008 and has overhangs compatible for GoldenBraid cloning.




Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 11
    Illegal NheI site found at 34
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 418
    Illegal XhoI site found at 199
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 101

Source

Synthesized by Twist Biosciences.

References

  • Choi, Y. J., Morel, L., Le François, T., Bourque, D., Bourget, L., Groleau, D., Massie, B., & Míguez, C. B. (2010). Novel, versatile, and tightly regulated expression system for Escherichia coli strains. Applied and environmental microbiology, 76(15), 5058–5066. https://doi.org/10.1128/AEM.00413-10