Difference between revisions of "Part:BBa K4207069"

 
 
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Sensor plasmid for detecting BYDV
 
Sensor plasmid for detecting BYDV
  
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===1. Usage and biology===
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===Usage and Biology===
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This sensor plasmid was designed to detect a conserved RNA sequence in the barley yellow dwarf virus (BYDV) genome. It should express the reporter protein mScarlet-I only in the presence of its specific trigger sequence. The translation is controlled by a toehold switch, which sequesters the RBS and the start codon in a stable RNA secondary structure. This structure is unfolded in the presence of the specific trigger, which is a conserved sequence in the BYDV genome.
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The translational activity can be measured by fluorescence intensity at 569/593 nm. The construct can be used in various reaction set ups. If used in a PURE (protein expression using recombinant elements) system or any other environment with reduced nuclease activity, it can be used as a linear plasmid. However, if used in a reaction with active nucleases, for instance a lysate-based cell-free expression system, the construct should be used as a circular plasmid to reduce degradation.
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K4207069 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K4207069 SequenceAndFeatures</partinfo>
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===2. Characterization===
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This part was not evaluated experimentally.
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===3. Conclusion===
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This part lacks experimental data. However, based on the modeling data of the toehold switch contained in this construct suggests that this part is not likely to detect BYDV genome.
  
  

Latest revision as of 16:08, 10 October 2022


Toehold sensor plasmid A95-mScarlet-I

Sensor plasmid for detecting BYDV

1. Usage and biology

This sensor plasmid was designed to detect a conserved RNA sequence in the barley yellow dwarf virus (BYDV) genome. It should express the reporter protein mScarlet-I only in the presence of its specific trigger sequence. The translation is controlled by a toehold switch, which sequesters the RBS and the start codon in a stable RNA secondary structure. This structure is unfolded in the presence of the specific trigger, which is a conserved sequence in the BYDV genome.

The translational activity can be measured by fluorescence intensity at 569/593 nm. The construct can be used in various reaction set ups. If used in a PURE (protein expression using recombinant elements) system or any other environment with reduced nuclease activity, it can be used as a linear plasmid. However, if used in a reaction with active nucleases, for instance a lysate-based cell-free expression system, the construct should be used as a circular plasmid to reduce degradation.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 171
    Illegal EcoRI site found at 219
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 171
    Illegal EcoRI site found at 219
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 171
    Illegal EcoRI site found at 219
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 171
    Illegal EcoRI site found at 219
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 171
    Illegal EcoRI site found at 219
  • 1000
    COMPATIBLE WITH RFC[1000]

2. Characterization

This part was not evaluated experimentally.

3. Conclusion

This part lacks experimental data. However, based on the modeling data of the toehold switch contained in this construct suggests that this part is not likely to detect BYDV genome.