Coding

Part:BBa_K5325001:Design

Designed by: Hien Le, Dang Minh Anh   Group: iGEM24_MichiganState   (2024-10-01)


Amy_plaA


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 637
    Illegal PstI site found at 71
    Illegal PstI site found at 519
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal SpeI site found at 637
    Illegal PstI site found at 71
    Illegal PstI site found at 519
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 637
    Illegal PstI site found at 71
    Illegal PstI site found at 519
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 637
    Illegal PstI site found at 71
    Illegal PstI site found at 519
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Included a C-terminal FLAG tag that helps with protein purification and Western Blot.
The DNA sequence was codon optimized for expression in E. coli and was found to work in Shewanella oneidensis.

Source

This part originated from the poly(L-lactic acid) depolymerase gene of Amycolatopsis sp. strain K104-1.
NCBI accession number: AB126228.1

References

Matsuda, E., Abe, N., Tamakawa, H., Kaneko, J., Kamio, Y. (2005). Gene Cloning and Molecular Characterization of an Extracellular Poly( l -Lactic Acid) Depolymerase from Amycolatopsis sp. Strain K104-1. J Bacteriol. 187(21):7333-7340