Coding

Part:BBa_K3279007:Design

Designed by: Liang Siwen   Group: iGEM19_CAU_China   (2019-10-15)


β-Glucosidase from Streptomyces coelicolor


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 287
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 427
    Illegal NgoMIV site found at 1103
    Illegal AgeI site found at 184
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 687
    Illegal BsaI.rc site found at 946


Design Notes

The primary concern was BioBrick compatibility so that illegal sites in RFC[10] did not appear in the sequence. And the sequeces was finally designed to link with pET30a(+) , so we added restriction enzyme cutting site EcoRI at the 5'end and HindIII at the 3'end. This part was expressed in E.coli BL21.


Source

The sequence is from Streptomyces coelicolor's genomic sequence.

References

[1]Gu, M. Z., Wang, J. C., Liu, W. B., Zhou, Y. & Ye, B. C. Expression and displaying of beta-glucosidase from Streptomyces coelicolor A3 in Escherichia coli. Applied biochemistry and biotechnology 170, 1713-1723, doi:10.1007/s12010-013-0301-4 (2013).