Regulatory

Part:BBa_K4159002:Design

Designed by: Melissa Hendrén   Group: iGEM22_Aalto-Helsinki   (2022-07-14)
Revision as of 17:25, 30 September 2022 by VeeraKurki (Talk | contribs) (→‎Design Notes)


T7 promoter


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 21
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 21
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 21
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The T7 promoter with the added stem-loop had been used in the plasmid (pBx1-VHH_template3-3XMyc-Spacer) [1], no additional changes were needed the reference sequence. The XbaI cutting site is in the stem-loop, and as the part was used in the DARPin construct fragment this was not an issue. This is a common consensus promoter that can be induced by IPTG. The following sequence after the promoter sequence is derived from the plasmid sequence used in the Chen et al. (2021) paper.

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