Composite

Part:BBa_K3580003:Design

Designed by: Kanta Suga   Group: iGEM20_Waseda   (2020-10-22)


Plux/tet-GFPssrA(LVA)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 745


Design Notes

In our project, we inserted this parts into pSB1C3 plasmid.


Source

We constructed this part by adding the ssrA-LVA tag sequence right after the GFP coding sequence of plux/tet-GFP(BBa_K934025)

References

[Gottesman et.al 1998] Gottesman S, Roche E, Zhou Y, Sauer RT. The ClpXP and ClpAP proteases degrade proteins with carboxy-terminal peptide tails added by the SsrA-tagging system. Genes Dev. 1998;12(9):1338-1347. doi:10.1101/gad.12.9.1338
[Shi et.al 2018] Shi X, Wu T, M Cole C, K Devaraj N, Joseph S. Optimization of ClpXP activity and protein synthesis in an E. coli extract-based cell-free expression system. Sci Rep. 2018;8(1):3488. Published 2018 Feb 22. doi:10.1038/s41598-018-21739-6
[Shin et.al 2010] Shin, J., & Noireaux, V. (2010). Study of messenger RNA inactivation and protein degradation in an escherichia coli cell-free expression system. Journal of Biological Engineering, 4(1), 9. doi:10.1186/1754-1611-4-9