Composite

Part:BBa_K2760031:Design

Designed by: iGEM18_TecMonterrey_GDL   Group: iGEM18_TecMonterrey_GDL   (2018-10-12)


SP2-Nsp2 Characterization construct L.rhamnosus


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 38
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 784
    Illegal AgeI site found at 896
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

    • Optimized for L.rhamnosus


Source

This plasmid expression vector has a spaCBA promoter, it contains a specific RBS for L. rhamnosus GG, the signal peptide SP2-Nsp2, M-cherry (BBa_K270029) as the fluorescent reporter with a His-Tag (6 histidines) attached to it; and the terminator 667.

This construct is meant to be used in characterizations of the signal peptide SP2-Nsp2 in E. coli, which would be excreted into the media by the Sec-dependent (Sec) secretion pathway, the general secretion E. coli route. When this signal peptides are linked to the N-terminus of recombinant proteins, they are critical for translocation and secretion of proteins in E. coli [1].

References

[1] Han, S., Machhi, S., Berge, M., Xi, G., Linke, T., & Schoner, R. (2017). Novel signal peptides improve the secretion of recombinant Staphylococcus aureus Alpha toxinH35L in Escherichia coli. AMB Express, 7, 93. http://doi.org/10.1186/s13568-017-0394-1