Tag

Part:BBa_K3781017:Design

Designed by: Nicolas Bayer   Group: iGEM21_TU_Kaiserslautern   (2021-10-08)


Strep8His, MocloMania B5


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

The sequence to this part was adapted from a Chlamy-optimized plasmid. By running a PCR on the sequence in question with specifically designed primers, we were able to generate a genetic part containing the sequence of interest flanked by invertedly oriented BbsI restriction sites. The overhangs did not have to be modified, because they were already B5 adapted. The restriction sites allowed us to introduce the amplified part into its respective plasmid backbone with a simple MoClo ligation.


Source

The genetic sequence to this part is based on a commercial, Chlamydomonas-adapted genetic part that is stored in the pCM0-099 plasmid of the Crozet MoClo Toolkit.[1] The Strep-tag II sequence itself is a synthetic peptide and doesn't have a natural genomic source. The 8His tag is a mere sequence of eight consecutive histidine amino acids and is not intentionally derived from any natural genomic source.


References

  1. Crozet et al. (2018) Birth of a Photosynthetic Chassis: A MoClo Toolkit Enabling Synthetic Biology in the Microalga Chlamydomonas reinhardtii. ACS Synthetic Biology 7:2074–2086