Tag

Part:BBa_K3781007:Design

Designed by: Nicolas Bayer   Group: iGEM21_TU_Kaiserslautern   (2021-10-07)


3xFLAG + HRV 3C Motif, MocloMania B3


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

This basic part was de novo synthesized and thus easily codon optimized towards Leishmania tarentolae.
We generated this part by designing it in silico to carry the sequence of interest flanked by two invertedly oriented BbsI recognition sites as well as the desired B3 overhangs. After commercial synthesis, this allowed us to introduce the part into its respective plasmid backbone with a simple MoClo ligation.


Source

The genetic sequence for this tag was directly derived from a publically available Addgene plasmid named pGCS-N3(3xFLAG), #85720.
Since the FLAG tag epitope is an artificially designed antigen, it doesn't have any genetic origin in nature. The HRV 3C motif sequence is based on the recognition sequence of a human rhinovirus protease and can thus be found in human genes, e.g. coding for transcription factors.[1]


References

  1. Amineva SP, Aminev AG, Palmenberg AC, Gern JE (October 2004). "Rhinovirus 3C protease precursors 3CD and 3CD' localize to the nuclei of infected cells". The Journal of General Virology. 85 (Pt 10): 2969–79. doi:10.1099/vir.0.80164-0. PMID 15448360