Coding

Part:BBa_K3349003:Design

Designed by: Kristi Turton   Group: iGEM20_Lethbridge_HS   (2020-09-23)
Revision as of 15:57, 6 October 2020 by Kristiturton (Talk | contribs) (→‎References)


Pectin Lyase Pnl-GFP


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The sequence was codon optimized for E. coli . The his tag was placed on the C-terminus. A TEV cut site was edded between the GFP and Pectase in case cleavage is needed after pruification.

Source

The GFP protein was taken from the uniprot database (P42212). The pectin lyase was also taken from the uniprot database and the sequence ID from NCBI is KAA8782864.1.

References

1. Keggi, C and Doran-Peterson, J. (2020) The homogalacturonan deconstruction system of Paenibacillus amylolyticus 27C64 requires no extracellular Pectin Methylesterase and has significant industrial potential. Applied and environmental microbiology. 86, e02275-19.

2.Roy, A., Kucukural, A., and Zhang, Y. (2010) I-TASSER: a unified platform for automated protein structure and function prediction. Nature Protocols, 5: 725-738.

3. Waterhouse, A., Bertoni, M., Bienert, S., Studer, G., Tauriello, G., Gumienny, R., Heer, F.T., de Beer, T.A.P., Rempfer, C., Bordoli, L., Lepore, R. and Schwede, T.(2018) SWISS-MODEL: homology modelling of protein structures and complexes. Nucleic Acids Res. 46, W296-W303.