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Part:BBa_K4159003:Design

Designed by: Melissa Hendrén   Group: iGEM22_Aalto-Helsinki   (2022-07-14)
Revision as of 17:26, 30 September 2022 by VeeraKurki (Talk | contribs) (→‎Design Notes)


Spacer 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

No changes were needed to the sequence. This sequence was taken from the pBx1-VHH_template3-3XMyc-Spacer [1], and placed at the end of the fragment for ribosome display of the DARPin construct. This spacer is derived from the pBx1-VHH_template3-3XMyc-Spacer plasmid from Chen et al. (2021). The spacer has been used in ribosome display for the selection of nanobodies. As our project is related to DARPin (design ankyrin repeat protein) selection, which is in the same size range (~14kDa) as the nanobodies, we were confident that the spacer was good for our selection. The spacer encodes for 72 amino acid residues, which should be long enough to “push out†the DARPin peptide structure from the ribosome.

Source

[1] Chen X, Gentili M, Hacohen N, Regev A. A cell-free nanobody engineering platform rapidly generates SARS-CoV-2 neutralizing nanobodies. Nat Commun. 2021 Sep 17;12(1):5506. doi: 10.1038/s41467-021-25777-z. PMID: 34535642; PMCID: PMC8448731.

References