Part:BBa_K3866003

Partial T5 Promoter - CuO- RBS GB compatible with A1-B2

Partial T5 promoter is a strong constitutive Promoter that can be used in the toxin-antitoxin system with the Cym repressor that our team uses in their system.

Fig.2:J23115 with RBS

Usage and Biology

T5 promoter has been used for many years for protein expression in E.coli. Partial T5 promoter is used with the downstream Operator CuO.

Design Notes

The sequence was domesticated . We removed BsmBI ,BsaI , BtgZI, BpiI sites in order to be compatible with GoldenBraid and MoClo. The sequence is cloned in pUPD2 BBa_K3505007 and has overhangs compatible for GoldenBraid cloning. This has position A1-B2.

Fig.1:The overhangs of this part in the GoldenBraid Grammar

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 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