Difference between revisions of "Part:BBa K3866016"
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[[File:T--Thessaly--AndersonRBS-snap.png|700px|thumb|none|<i><b>Fig.2:</b>J23115 with RBS</i>]]
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[[File:T--Thessaly--Plac.MazE.TT.png|700px|thumb|none|<i><b>Fig.2:</b>Tha Level A of MazE antitoxin with the Lactose promoter and a double terminator in an alpha2 vector.</i>]]
  
 
===Usage and Biology===
 
===Usage and Biology===
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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
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*Douglas F. Browning, Rita E. Godfrey, Kirsty L. Richards, Colin Robinson, Stephen J.W. Busby; Exploitation of the Escherichia coli lac operon promoter for controlled recombinant protein production. Biochem Soc Trans 30 April 2019; 47 (2): 755–763. doi: https://doi.org/10.1042/BST20190059
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*Choi, W., Yamaguchi, Y., Park, J.-Y., Park, S.-H., Lee, H.-W., Lim, B.-K., … Park, J.-H. (2021). Functional Characterization of the mazEF Toxin-Antitoxin System in the Pathogenic Bacterium Agrobacterium tumefaciens. Microorganisms, 9(5), 1107. doi:10.3390/microorganisms9051107

Latest revision as of 10:42, 29 September 2021


Plac-MazE-double terminator


Fig.2:Tha Level A of MazE antitoxin with the Lactose promoter and a double terminator in an alpha2 vector.

Usage and Biology

This part includes the weak promoter of the lactose operon without the lactose operator,which is used for protein expression. Also, it includes the antitoxin MazE that will be utilized to the toxin-antitoxin system and the double terminator.



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 alpha 2 vector BBa_K3505009 and has overhangs compatible for GoldenBraid cloning.



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 105
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

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
  • Douglas F. Browning, Rita E. Godfrey, Kirsty L. Richards, Colin Robinson, Stephen J.W. Busby; Exploitation of the Escherichia coli lac operon promoter for controlled recombinant protein production. Biochem Soc Trans 30 April 2019; 47 (2): 755–763. doi: https://doi.org/10.1042/BST20190059
  • Choi, W., Yamaguchi, Y., Park, J.-Y., Park, S.-H., Lee, H.-W., Lim, B.-K., … Park, J.-H. (2021). Functional Characterization of the mazEF Toxin-Antitoxin System in the Pathogenic Bacterium Agrobacterium tumefaciens. Microorganisms, 9(5), 1107. doi:10.3390/microorganisms9051107