Composite

Part:BBa_K4653114:Design

Designed by: Tangkun Zhu   Group: iGEM23_SZU-China   (2023-10-10)


pMA5 - Veg promoter - SpoVG RBS - Epr - BvEP - 2x fd terminator


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 NgoMIV site found at 926
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1414
    Illegal BsaI.rc site found at 1482


Design Notes

We synthesized the nucleic acid fragment of Veg promoter-SpoVG RBS-Epr signal peptide-BvEP-T7TE Terminator and designed homologous arm primers required for seamless cloning. The plan is to link the fragment with plasmid vector pMA5 through seamless cloning, and transfer it to Bacillus subtilis WB800 to verify follow-up experiments.


Source

We synthesized the nucleic acid fragment of Veg promoter-SpoVG RBS-Epr signal peptide-BvEP-T7TE Terminator and designed homologous arm primers required for seamless cloning. The plan is to link the fragment with plasmid vector pMA5 through seamless cloning, and transfer it to Bacillus subtilis WB800 to verify follow-up experiments.

References

[1] Li Z, Hu J, Sun Q, Zhang X, Chang R, Wang Y. A novel elicitor protein phosphopentomutase from Bacillus velezensis LJ02 enhances tomato resistance to Botrytis cinerea. Front Plant Sci. 2022 Nov 29;13:1064589. doi: 10.3389/fpls.2022.1064589.
[2] Daudi, A. and O’Brien, J. A. (2012). Detection of Hydrogen Peroxide by DAB Staining in Arabidopsis Leaves. Bio-protocol 2(18): e263. DOI: 10.21769/BioProtoc.263.
[3] Pruitt RN, Gust AA, Nürnberger T. Plant immunity unified. Nat Plants. 2021 Apr;7(4):382-383. doi: 10.1038/s41477-021-00903-3.
[4] Yuan M, Ngou BPM, Ding P, Xin XF. PTI-ETI crosstalk: an integrative view of plant immunity. Curr Opin Plant Biol. 2021 Aug;62:102030. doi: 10.1016/j.pbi.2021.102030. Epub 2021 Mar 5.
[5] Hu J, Chang R, Yuan Y, Li Z, Wang Y. Identification of Key Residues Essential for the Activation of Plant Immunity by Subtilisin From Bacillus velezensis LJ02. Front Microbiol. 2022 Aug 15;13:869596. doi: 10.3389/fmicb.2022.869596.