Coding

Part:BBa_K2238002:Design

Designed by: Bryan Wilkins   Group: iGEM17_ManhattanCol_Bronx   (2017-10-21)


Glucose oxidase (A. niger) Cysteine tagged


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 36
    Illegal BamHI site found at 346
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 370
    Illegal BsaI.rc site found at 1588


Design Notes

The amino acid sequence was converted to the E. coli optimized DNA coding sequence (IDT https://www.idtdna.com/CodonOpt). We added the coding sequence for an N-terminal His-tag and TEV protease for convenient protein isolation. This leaves a small glycine scar on the N-terminal end of the protein following enzymatic cleavage. There is a short 4-glycine-serine tag followed by a 3X cysteine tag site prior to the stop codon. The cysteines were added as an affinity tag for gold nanoparticles/wires.


This parts coding is based on part BBa_K2238000 with identical sequencing prior to the CYS-tag addition.

Source

Aspergillus niger Gene Bank: AAA32695.1 (E.C 1.1.3.4)

References

Frederick, K.R., Tung, J., Emerick, R.S., Masiarz, F.R., Chamberlain, S.H., Vasavada, A., Rosenberg, S., Chakraborty, S., Schopfer, L.M., Schopter, L.M. et al. Glucose oxidase from Aspergillus niger. Cloning, gene sequence, secretion from Saccharomyces cerevisiae and kinetic analysis of a yeast-derived enzyme. J. Biol. Chem. 1990, vol 265 (7) pp. 3793-3802.