Coding

Part:BBa_K3014004:Design

Designed by: Benedikt Schober, Jan Müller, Kai Schülke   Group: iGEM19_Stuttgart   (2019-10-20)


Vibrio natriegens tRNA TCC


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design

This part is cloned into the ptRNA backbone (BBa_K3014006).The tRNA backbone uses a p15A ori, as it also used in the established tRNA vector ‘pRARE’ from Merck Inc., which is used in the famous E. coli strain Rosetta.5 By using this Ori it is possible to express two plasmids simultaneously in V. natriegens. This allows the level of tRNAs to be increased, thus enhancing protein expression. As selection markers, we chose tetracycline as Vibrio natriegens is not majorly susceptible to beta-lactamase inhibiting antibiotics (e.g. ampicillin, kanamycin, ...), as advised by the iGEM team Marburg 2018.


Source

All parts have been synthesized by Integrated DNA Technologies.