Difference between revisions of "Part:BBa K4159002:Design"
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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. | 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. | ||
| + | The part has been used for ribosome display as well as the promoter for GFP binding DARPin expression. Check out the parts BBa_K4159012 and BBa_K4159000 for a more detailed description. | ||
===Source=== | ===Source=== | ||
Latest revision as of 16:26, 1 October 2022
T7 promoter
- 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. 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.
The part has been used for ribosome display as well as the promoter for GFP binding DARPin expression. Check out the parts BBa_K4159012 and BBa_K4159000 for a more detailed description.
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.