Composite
lon sensor

Part:BBa_K3773523:Design

Designed by: Justin Berg   Group: iGEM21_William_and_Mary   (2021-10-15)


Circuit to report Plon 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 459
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We found that of various protease genes, lon is one of the most frequently differentially expressed in E. coli upon transformation with a heterologous circuit. As a result, we use the lon promoter as one representative of promoters related to protease expression.

The sequence inputted as a scar after the RBS is a spacer which has been shown to allow for expression with BBa_B0034 and sfGFP1.

UNS 1 and UNS 10 flank this part in order to allow for easy Gibson assembly as detailed by Torella et al., 20142.

Source

See basic parts

References

1Clifton, 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.

2Torella, 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.