Composite

Part:BBa_K1357007:Design

Designed by: Matthew Tucker   Group: iGEM14_Virginia   (2014-10-09)

NylC - Nylon Degradation Construct


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 115
    Illegal AgeI site found at 263
    Illegal AgeI site found at 335
    Illegal AgeI site found at 521
    Illegal AgeI site found at 719
    Illegal AgeI site found at 845
  • 1000
    COMPATIBLE WITH RFC[1000]

The construct includes a high expression level promoter (J23110), a high expression level ribosomal binding site (B0034) and a double terminator (B0015).

Design Notes

To obtain the coding sequence for nylC we found the sequence in genbank and then optimized it for E. coli expression. Subsequently the pelB secretion tagged was affixed to the beginning of the coding sequence for nylC.

The nylC DNA was synthesized by IDT.

Source

Flavobacterium sp. plasmid pOAD2

References

[http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3281642/]Negoro, S. et al. "Three-dimensional Structure of Nylon Hydrolase and Mechanism of Nylon-6 Hydrolysis". J Biol Chem. Feb 10, 2012; 287(7): 5079–5090.

[1] pelB secretion tag registry entry.