Regulatory
Part:BBa_K4159002:Design
Designed by: Melissa Hendrén Group: iGEM22_Aalto-Helsinki (2022-07-14)
T7 promoter
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 21
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 21
- 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 21
- 1000COMPATIBLE 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.
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.