We used two parts for our experiments. Both constructs were assembled using NEB Gibson Assembly Master Mix E2611.
The part shown in the left panel consists of the Pyrococcus furiosus DNA polymerase insert gene in the JOE vector. The DNA polymerase is codon-optimized. This part expresses the P. furiosus DNA polymerase enzyme when transformed in DH5α. The P. furiosus DNA polymerase enzyme is fused to a single- stranded binding protein. This gives the enzyme characteristics similar to the commercial Phusion DNA polymerase. The sequence for this part could not be verified and was not sent to iGEM HQ.
The part shown in the right panel consists of the Thermotoga maritima DNA ligase insert gene in the JOE vector. This part expresses the T. maritima DNA ligase enzyme when transformed in DH5α. The sequence for this part was successfully verified and was sent to iGEM HQ.The registry link is as follows: http://parts.igem.org/Part:BBa_K2468000
Both parts have indicated potential for behaving as expected, however, we are conducting more experiments to obtain concrete results regarding the behavior of each plasmid construct. One experiment has indicated the successful behavior of the P. furiosus DNA polymerase enzyme.