Difference between revisions of "Part:BBa K4765129"
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| − | | ''' | + | | '''Figure 1. Sequencing result of nsl (no stem-loop before Twister ribozyme cleavage site).''' Sanger sequencing verified we have removed the stem-loop before ribozyme sequence. We also construct plasmids with stem-loop new2, new6, new10, bad2, bad6, bad10, all of which were designed by our [https://2023.igem.wiki/fudan/software Software RAP], and fully characterized using functional assays. |
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==Reference== | ==Reference== | ||
Revision as of 23:55, 10 October 2023
stem-loop test
Introduction
This compoiste part is a stem-loop-deleted version of BBa_K4765120, which includes StayGold and mScarlet. Its red-green fluorescence intensity ratio can be compared with BBa_K4765120 to assess the stem-loop's ability to prevent mRNA degradation.
Usage and Biology
We use this composite part to test the following stem-loops'[1] ability to prevent mRNA degradation.
nsl: 5-AAACACCCACCACAAUUUCCACCGUUU UUUGU-3 liu2023: 5-AAACACCCACCACAAUUUCCACCGUUU CCCGACGCUUCGGCGUCGGG UUUGU-3 new2: 5-AAACACCCACCACAAUUUCCACCGUUU CCCCGUCGGCUGCU UUUGU-3 new6: 5-AAACACCCACCACAAUUUCCACCGUUU AGACGCUCGGCGUCCU UUUGU-3 new10: 5-AAACACCCACCACAAUUUCCACCGUUU ACUGGGGGGAUCGAGGUCUUU UUUGU-3 bad2: 5-AAACACCCACCACAAUUUCCACCGUUU GCCGAUCGGGU UUUGU-3 bad6: 5-AAACACCCACCACAAUUUCCACCGUUU AGACGCUCGGCGUCCU UUUGU-3 bad10: 5-AAACACCCACCACAAUUUCCACCGUUU GGCGGCGCUACAGCGUCGU UUUGU-3
Characterization
Sequencing map
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| Figure 1. Sequencing result of nsl (no stem-loop before Twister ribozyme cleavage site). Sanger sequencing verified we have removed the stem-loop before ribozyme sequence. We also construct plasmids with stem-loop new2, new6, new10, bad2, bad6, bad10, all of which were designed by our Software RAP, and fully characterized using functional assays. |
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 1389
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 700
Illegal BsaI.rc site found at 720
Reference
- ↑ Liu, Y., Wu, Z., Wu, D., Gao, N., & Lin, J. (2022). Reconstitution of Multi-Protein Complexes through Ribozyme-Assisted Polycistronic Co-Expression. ACS Synthetic Biology, 12(1), 136–143. https://doi.org/10.1021/acssynbio.2c00416