Part:BBa_K2091008:Design

LC-Cutinase Variant R7

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 394
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 195
  Illegal BamHI site found at 597
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 550
  
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

The sequence was designed using the PROSS algorithm¹ and the LC-Cutinase tertiary structure². It was synthesized by Gen9. One point mutation was introduced to remove a PstI cut site.

Source

This is a mutant of the LC-Cutinase protein³, the original sequence was acquired from the 2012 UC-Davis iGEM team. (BBa_K936000)

References

1. Goldenzweig, A., Goldsmith, M., Hill, S. E., Gertman, O., Laurino, P., Ashani, Y., ... & Lieberman, R. L. (2016). Automated structure-and sequence-based design of proteins for high bacterial expression and stability. Molecular Cell, 63(2), 337-346.
2. Sulaiman, S., You, D. J., Kanaya, E., Koga, Y., & Kanaya, S. (2014). Crystal structure and thermodynamic and kinetic stability of metagenome-derived LC-cutinase. Biochemistry, 53(11), 1858-1869.‏
3. Sulaiman, S., Yamato, S., Kanaya, E., Kim, J. J., Koga, Y., Takano, K., & Kanaya, S. (2012). Isolation of a novel cutinase homolog with polyethylene terephthalate-degrading activity from leaf-branch compost by using a metagenomic approach. Applied and environmental microbiology, 78(5), 1556-1562.‏