Plasmid

Part:BBa_K4212044:Design

Designed by: Yuancheng Ding   Group: iGEM22_Imperial_College_London   (2022-09-30)


SDP1


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 163
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

PlepA is constitutive promoter, which is responsible for the expression of protein PlepA and important for the transcription of the bicistronic operon (Homuth et al., 1996). PlepA can improve the fidelity of translation by regulating the mRNA-tRNA interactions(Qin et al., 2006). Previous iGEM team iGEM12_LMU-Munich used this promoter before. The use of the Spy terminator by the previous iGEM team iGEM18_William_and_Mary inspired us to use this terminator. According to the website Explora Biotech, the Spy terminator is a "fairly strong palindromic rho-independent transcriptional terminator". We included the gRNA in the Golden Gate assembly as a part of our self-digesting plasmid.


Source

Synthetic construct

References

[1] https://parts.igem.org/Part:BBa_K823002 [2] Homuth, G., Heinemann, M., Zuber, U. & Schumann, W. 1996 (n.d.) The genes lepA and hemN form a bicistronic operon in Bacillus subtilis. Microbiology. 142 (7), 1641–1649. doi:10.1099/13500872-142-7-1641. [3] Qin, Y., Polacek, N., Vesper, O., Staub, E., Einfeldt, E., Wilson, D.N. & Nierhaus, K.H. (2006) The Highly Conserved LepA Is a Ribosomal Elongation Factor that Back-Translocates the Ribosome. Cell. 127 (4), 721–733. doi:10.1016/j.cell.2006.09.037. [4] https://parts.igem.org/Part:BBa_K2680400 [5] https://doulix.com/biomodules/GNO9X1/