Difference between revisions of "Part:BBa K4765129"

Line 12: Line 12:
 
We use this composite part to test the following stem-loops' ability to prevent mRNA degradation.
 
We use this composite part to test the following stem-loops' ability to prevent mRNA degradation.
 
nsl:  <pre>5-AAACACCCACCACAAUUUCCACCGUUU UUUGU-3</pre>
 
nsl:  <pre>5-AAACACCCACCACAAUUUCCACCGUUU UUUGU-3</pre>
liu2023 <ref>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</ref>: <pre> 5-AAACACCCACCACAAUUUCCACCGUUU CCCGACGCUUCGGCGUCGGG UUUGU-3</pre>
+
liu2023 <ref>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</ref>: <pre>5-AAACACCCACCACAAUUUCCACCGUUU CCCGACGCUUCGGCGUCGGG UUUGU-3</pre>
new2:  <pre> 5-AAACACCCACCACAAUUUCCACCGUUU CCCCGUCGGCUGCU UUUGU-3</pre>
+
new2:  <pre>5-AAACACCCACCACAAUUUCCACCGUUU CCCCGUCGGCUGCU UUUGU-3</pre>
 
new6:  <pre>5-AAACACCCACCACAAUUUCCACCGUUU AGACGCUCGGCGUCCU UUUGU-3</pre>
 
new6:  <pre>5-AAACACCCACCACAAUUUCCACCGUUU AGACGCUCGGCGUCCU UUUGU-3</pre>
new10:  <pre> 5-AAACACCCACCACAAUUUCCACCGUUU ACUGGGGGGAUCGAGGUCUUU UUUGU-3</pre>
+
new10:  <pre>5-AAACACCCACCACAAUUUCCACCGUUU ACUGGGGGGAUCGAGGUCUUU UUUGU-3</pre>
 
bad2:  <pre>5-AAACACCCACCACAAUUUCCACCGUUU GCCGAUCGGGU UUUGU-3</pre>
 
bad2:  <pre>5-AAACACCCACCACAAUUUCCACCGUUU GCCGAUCGGGU UUUGU-3</pre>
bad6:  <pre> 5-AAACACCCACCACAAUUUCCACCGUUU AGACGCUCGGCGUCCU UUUGU-3</pre>
+
bad6:  <pre>5-AAACACCCACCACAAUUUCCACCGUUU AGACGCUCGGCGUCCU UUUGU-3</pre>
 
bad10:  <pre>5-AAACACCCACCACAAUUUCCACCGUUU GGCGGCGCUACAGCGUCGU UUUGU-3</pre>
 
bad10:  <pre>5-AAACACCCACCACAAUUUCCACCGUUU GGCGGCGCUACAGCGUCGU UUUGU-3</pre>
  

Revision as of 12:37, 9 October 2023


stem-loop test contributed by Fudan iGEM 2023


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' ability to prevent mRNA degradation.

nsl:
5-AAACACCCACCACAAUUUCCACCGUUU UUUGU-3
liu2023 [1]:
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

contributed by Fudan iGEM 2023
Figure1 Sequencing map of stem-loop test Sanger sequencing verified we have removed the stem-loop before ribozyme sequence.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 1389
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 700
    Illegal BsaI.rc site found at 720


Reference

  1. 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