Part:BBa_K3866005

Plac-RBS GB compatible with A1-B2

Promoter lac is a weak inducible promoter from the lactose operon. This part has no lactose operator, because our team needed only the promoter.

Fig.2:Lactose Promoter with RBS and compatible ends for Golden Braid Cloning without Lactose operator.

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 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 pUPD2 BBa_K3505007 and has overhangs compatible for GoldenBraid cloning. The Promoter 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