Composite
Part:BBa_K1357007:Design
Designed by: Matthew Tucker Group: iGEM14_Virginia (2014-10-09)
NylC - Nylon Degradation Construct
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE 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 - 1000COMPATIBLE 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.