Part:BBa_K3866020

J23115-CymR-Stuffer a2

Fig.2:The omega 2 vector with two translational units. The one is J23115-CymR-double terminator and the other one is the stuffer.

Usage and Biology

This composite part is consisted of the translational unit of the strong constitutive promoter J23115 BBa_K3505012and the Cym Repressor BBa_K415202and the transcriptional unit with the stuffer BBa_K3866019, which is a part of DNA non-coding with a length close to 150 basepairs. The stuffer is used so that we can swap the insert from the alpha 2 vector into the omega 2 vector BBa_K3505011.

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 omega2 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

• Sarrion-Perdigones A, Falconi EE, Zandalinas SI, Juárez P, Fernández-del-Carmen A, et al. (2011) GoldenBraid: An Iterative Cloning System for Standardized Assembly of Reusable Genetic Modules. PLOS ONE 6(7): e21622. https://doi.org/10.1371/journal.pone.0021622
• 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