Difference between revisions of "Part:BBa K4159002:Design"

 
 
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===Design Notes===
 
===Design Notes===
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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.
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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.
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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


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.

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.

References