Composite

Part:BBa_K3511002:Design

Designed by: Yung Mau SO   Group: iGEM20_Hong_Kong_CityU   (2020-10-08)


Papain G23W + PueA R392F + PETase


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 3659
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

A glycine to tryptophan mutation at amino acid position 23 was created in papain (polyurethanase) and an arginine to phenylalanine mutation at amino acid position 392 was created in PueA (polyurethanase).

Source

T7 promoter: T7 phage, sequence found in https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3592457/

RBS: https://parts.igem.org/Part:BBa_B0034

Papain G23W: Carica papaya, sequence was originated from https://www.ncbi.nlm.nih.gov/nuccore/M15203.1?report=genbank

GSG linker+P2A: https://parts.igem.org/Part:BBa_K1537016

PueA R392F: Pseudomonas chlororaphis, sequence originated from https://www.ncbi.nlm.nih.gov/nuccore/AF069748.1?report=fasta

GSG linker+T2A: https://parts.igem.org/Part:BBa_K1537017

PETase: https://parts.igem.org/Part:BBa_K2013002

References

Amrani, A., Barakate, A., Askari, B., Li, X., Roberts, A., Ryan, M., & Halpin, C. (2004, May 01). Coordinate expression and independent subcellular targeting of multiple proteins from a single transgene. Retrieved September 13, 2020, from http://www.plantphysiol.org/content/135/1/16.long

Joo, S., Cho, I., Seo, H., Son, H., Sagong, H., Shin, T., . . . Kim, K. (2018, January 26). Structural insight into molecular mechanism of poly(ethylene terephthalate) degradation. Retrieved October 21, 2020, from https://www.nature.com/articles/s41467-018-02881-1

Kim, J., Lee, S., Li, L., Park, H., Park, J., Lee, K., . . . Choi, S. (2011). High cleavage efficiency of a 2A peptide derived from porcine teschovirus-1 in human cell lines, zebrafish and mice. Retrieved September 28, 2020, from https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3084703/

Paul, S., Stang, A., Lennartz, K., Tenbusch, M., & Überla, K. (2012, October 15). Selection of a T7 promoter mutant with enhanced in vitro activity by a novel multi-copy bead display approach for in vitro evolution. Retrieved September 13, 2020, from https://academic.oup.com/nar/article-lookup/doi/10.1093/nar/gks940

Phua, S., Castillo, E., Anderson, J., & Hiltner, A. (2004, September 13). Biodegradation of a polyurethane in vitro. Retrieved September 13, 2020, from https://onlinelibrary.wiley.com/doi/abs/10.1002/jbm.820210207

Szymczak-Workman, A. L., Vignali, K. M., & Vignali, D. A. (2012). Design and construction of 2A Peptide-linked multicistronic vectors. Cold Spring Harbor Protocols, 2012(2). doi:10.1101/pdb.ip067876

Yoshida, S., Hiraga, K., Takehana, T., Taniguchi, I., Yamaji, H., Maeda, Y., . . . Oda, K. (2016, March 11). A bacterium that degrades and assimilates poly(ethylene terephthalate). Retrieved October 02, 2020, from https://science.sciencemag.org/content/351/6278/1196.long