Composite

Part:BBa_K4204020:Design

Designed by: Jianfei Song   Group: iGEM22_BNDS_China   (2022-10-11)


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

[3] Lee, K. O., Kong, M., Kim, I., Bai, J., Cha, S., Kim, B., Ryu, K.-S., Ryu, S., & Suh, J.-Y. (2019). Structural basis for cell-wall recognition by bacteriophage PBC5 endolysin. Structure, 27(9). https://doi.org/10.1016/j.str.2019.07.001