Coding

Part:BBa_K4580000:Design

Designed by: Michael Constant   Group: iGEM23_Cornell   (2023-10-09)

Single Nucleotide Polymorphism (SNP)

To maintain compatibility with the iGEM Parts Registry, a SNP was done at base 480 and maintains the same amino acid sequence.


Enzyme Isolation

To allow for easy enzyme isolation, a 6x-histidine tag was added to the NdmA. This design choice allowed Team Cornell to use Ni-NTA Affinity Chromatography to isolate our enzymes of interest for further encapsulation into our reactor.


Source

NdmA, NdmB, and, NdmD were amplified from the bacteria Pseudomonas putida CBB5 genomic DNA.


References

Summers, R. M., Louie, T. M., Yu, C. L., Gakhar, L., Louie, K. C., & Subramanian, M. (2012). Novel, highly specific N-demethylases enable bacteria to live on caffeine and related purine alkaloids. Journal of bacteriology, 194(8), 2041–2049. https://doi.org/10.1128/JB.06637-11

UT Austin. (2012). BBA_K734000. NdmABCD operon. https://parts.igem.org/Part:BBa_K734000