Coding

Part:BBa_K3682004:Design

Designed by: YOU-CHENG LEE   Group: iGEM20_NYMU-Taipei   (2020-10-15)


cleavage site 1 on spike protein sequence


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 sequence does not include a start codon and a stop codon, so we have to add a start codon atg at the top. As for the stop codon, we put a TEV cleavage site and a 6xhistag with a stop codon in the back for later protein purification. We also need a bacterial expression vector(part BBa_K3682000) with a ribosome binding site to express the polypeptide in BL21 strain.


Fig1. The structure of spike cut1 fragment. The cutting site is labeled in red.

Source

We cloned the sequence with PCR from SARS CoV2 spike protein plasmid of addgene.org(id #145730)

References

1. addgene website: https://www.addgene.org/145730/