Part:BBa_K3773522:Design

Circuit to report ftsLp2 expression in vivo

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 291
  
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

We hoped to consider the effect of a heterologous circuit on a host's capacity to carry out post-translational modifications, and looked into glycosylation as a more specific target. This circuit places sfGFP after the ftsLp2 promoter in order to report its expression; this promoter normally controls genes murG and murC, which are involved in peptidoglycan synthesis¹.

The sequence inputted as a scar after the RBS is a spacer which has been shown to allow for expression with BBa_B0034 and sfGFP².

UNS 1 and UNS 10 flank this part in order to allow for easy Gibson assembly as detailed by Torella et al., 2014³.

Source

See basic parts.

References

¹Barreteau, H., Kovac, A., Boniface, A., Sova, M., Gobec, S., & Blanot, D. (2008). Cytoplasmic steps of peptidoglycan biosynthesis. FEMS Microbiology Reviews, 32(2), 168–207

²Clifton, K. P., Jones, E. M., Paudel, S., Marken, J. P., Monette, C. E., Halleran, A. D., ... & Saha, M. S. (2018). The genetic insulator RiboJ increases expression of insulated genes. Journal of biological engineering, 12(1), 1-6.

³Torella, J. P., Boehm, C. R., Lienert, F., Chen, J. H., Way, J. C., & Silver, P. A. (2014). Rapid construction of insulated genetic circuits via synthetic sequence-guided isothermal assembly. Nucleic acids research, 42(1), 681-689.