Tag

Part:BBa_K2909001:Design

Designed by: Laurent CHEN   Group: iGEM19_Sorbonne_U_Paris   (2019-08-26)


HiBiT-B5 MoClo C. reinhardtii


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 Notes

We designed the HiBiT tag to be standardized to the Phytobrick standard for the C. reinhardtii MoClo Kit (Crozet et al. 2018).
BbsI sites and fusion sites corresponding to the B5 position (C-terminal tag) were added on both side to be compatible with the MoClo assembly standard.
A stop codon (TAA) was added at the 3' end of the HiBiT tag as it is a C-terminal tag.

Source

This part comes from Promega (Schwinn et al. 2018).

References

  1. Schwinn, M. K. et al. CRISPR-Mediated Tagging of Endogenous Proteins with a Luminescent Peptide. ACS Chem. Biol. 13, 467–474 (2018).
  2. Crozet, P. et al. Birth of a Photosynthetic Chassis: A MoClo Toolkit Enabling Synthetic Biology in the Microalga Chlamydomonas reinhardtii. ACS Synth. Biol. 7, 2074–2086 (2018).
  3. 1.Weber, E., Engler, C., Gruetzner, R., Werner, S. & Marillonnet, S. A Modular Cloning System for Standardized Assembly of Multigene Constructs. PLoS ONE 6, e16765 (2011).