Coding

Part:BBa_K4447004:Design

Designed by: Rafael Garcia Gomez   Group: iGEM22_TecMonterrey_GDL   (2022-09-29)

FRET-based system for the detection of erythromycin


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 1913
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2562

Design Notes

A gly-gly-ser spacer, and a polyhistidine-tag were added before stop codon for protein purification. NcoI and XhoI restriction sites were added in 5' and 3' ends for protein overexpression in pBAD/Myc-His plasmids. There are no scars between each protein.

Source

The original sequence was reported by Stassi et al. (1993). Sequence can be obtained through GenBank (L05776). The sequence for BBa_K1159302 was reported by Poëa-Guyon et al. (2013). The sequence for BBa_K1907000 was reported by Nagai et al. (2002).

References

[1]. Nagai, T., Ibata, K., Park, E. S., Kubota, M., Mikoshiba, K., & Miyawaki, A. (2002). A variant of yellow fluorescent protein with fast and efficient maturation for cell-biological applications. Nature Biotechnology, 20(1), 87–90. doi:10.1038/nbt0102-87

[2]. Poëa-Guyon, S., Pasquier, H., Mérola, F., Morel, N., & Erard, M. (2013). The enhanced cyan fluorescent protein: a sensitive pH sensor for fluorescence lifetime imaging. Analytical and bioanalytical chemistry, 405(12), 3983–3987. https://doi.org/10.1007/s00216-013-6860-y

[3]. Stassi, D., Donadio, S., Staver, M. J., & Katz, L. (1993). Identification of a Saccharopolyspora erythraea gene required for the final hydroxylation step in erythromycin biosynthesis. Journal of bacteriology, 175(1), 182–189. https://doi.org/10.1128/jb.175.1.182-189.1993