Part:BBa_K4204020:Design

Crystal violet induced GFP improved with ProQC system

Assembly Compatibility:

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

  Illegal NheI site found at 1166
  Illegal NheI site found at 1248
  Illegal NheI site found at 2332
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 1045
  Illegal BamHI site found at 1943
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 475
  Illegal NgoMIV site found at 1995
  Illegal AgeI site found at 625
  
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

The initial design of the part has a flaw (a gap between the RBS and the start codon leads to a frameshift of the resulting protein). This problem is fixed now by another round of design, build and test.

Source

The protein quality control system is from Yang et al's work [1], the crystal violet induction system is from Ruegg et al.'s work [2], and the CDS of LysPBC5 is from Lee et al's work [3]. Assembled by BNDS_China 2022.

References

[1] Yang, J., Han, Y.H., Im, J. et al. Synthetic protein quality control to enhance full-length translation in bacteria. Nat Chem Biol 17, 421–427 (2021). https://doi.org/10.1038/s41589-021-00736-3

[2] Ruegg, T. L., Pereira, J. H., Chen, J. C., DeGiovanni, A., Novichkov, P., Mutalik, V. K., Tomaleri, G. P., Singer, S. W., Hillson, N. J., Simmons, B. A., Adams, P. D., & Thelen, M. P. (2018). Jungle Express is a versatile repressor system for tight transcriptional control. Nature Communications, 9(1). https://doi.org/10.1038/s41467-018-05857-3