All biobricks were synthesized by Integrated DNA Technologies.
In total, we used ten parts (nine basic parts and one composite part) and submitted them to the registry. Among them, five parts are tRNA genes from Vibrio natriegens which should be used to increase the levels of rare tRNAs in Vibrio natriegens. These tRNA genes should be cloned individually into a self-designed plasmid backbone. To increase the tRNA levels of all tRNAs simultaneously, a combined tRNA fragment was made.
Since the measurement of single tRNA species is complicated and not established, a qPCR method was developed. Using an adapter, specific tRNA levels could be measured. This adapter is available below.
Additionally, the Biobricks GFP (http://parts.igem.org/Part:BBa_E0040) and sfGFP (http://parts.igem.org/Part:BBa_I746916) have been improved by optimizing them for Vibrio natriegens. The sequences were analyzed for rare codons and then exchanged with more frequently used ones. This means that no classical codon optimization was performed, but rare codons, which could have a strong influence on the expression strength, were exchanged. By doing so, we wanted to increase the expression of GFP and sfGFP in Vibrio natriegens and thus provide a better measurement tool.
|1||BBa_K3014000||Coding||Vibrio natriegens tRNA AGA||Benedikt Schober, Jan Müller, Kai Schülke||135|
|2||BBa_K3014001||Coding||Vibrio natriegens tRNA AGG||Benedikt Schober, Jan Müller, Kai Schülke||135|
|3||BBa_K3014002||Coding||Vibrio natriegens tRNA CGG||Benedikt Schober, Jan Müller, Kai Schülke||160|
|4||BBa_K3014003||Coding||Vibrio natriegens tRNA TGC||Benedikt Schober, Jan Müller, Kai Schülke||161|
|5||BBa_K3014004||Coding||Vibrio natriegens tRNA TCC||Benedikt Schober, Jan Müller, Kai Schülke||151|
|6||BBa_K3014006||Plasmid_Backbone||ptRNA_backbone||Benedikt Schober, Jan Müller, Kai Schülke||2161|
|7||BBa_K3014007||Coding||Codon optimized GFP for Vibrio natriegens||Benedikt Schober||718|
|8||BBa_K3014008||Coding||Codon optimized sfGFP for Vibrio natriegens||Benedikt Schober||718|
|9||BBa_K3014009||Measurement||DNA-RNA-Hybrid-Adapter for tRNA quantification||Katharina Hofer, Sini Münßinger, Jan Seeger, Gabriele Kepp||130|
|10||BBa_K3014010||Terminator||Terminator of rrnA operon V. natriegens||Jan Müller, Kai Schülke||54|
|11||K3014011||Regulatory||V. natriegens rrnA P1 Promoter||Jan Müller, Kai Schülke||71|
|12||BBa_K3014012||Coding||Vibrio natriegens tRNA construct||Benedikt Schober, Jan Müller, Kai Schülke||915|