Difference between revisions of "Part:BBa K4244023"
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<partinfo>BBa_K4244023 short</partinfo>
 
<partinfo>BBa_K4244023 short</partinfo>
  
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The pcspA (<partinfo>BBa_K4244021</partinfo>) is a constitutive promoter that has a high rate of transcription but contains a long 5′ UTR of 159 bp that, at 37°C, acquires an unstable secondary structure and leads to its degradation. On the other hand, at lower temperatures it forms a stable configuration, which allows translation. In this case, a toxin (ccdB) under the cspA promoter is expressed but counteracted by the antitoxin ccdA (<partinfo>BBa_K4244023</partinfo>), which in turn is expressed by being placed after the constitutive LacUV5 (<partinfo>BBa_M36801</partinfo>) (1). At lower temperature the cspA stabilizes and ccdB overexpresses ccdA, which leads to cell death.  
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The pcspA (<partinfo>BBa_K4244021</partinfo>) is a constitutive promoter that has a high rate of transcription but contains a long 5′ UTR of 159 bp that, at 37°C, acquires an unstable secondary structure and leads to its degradation. On the other hand, at lower temperatures it forms a stable configuration, which allows translation. In this case, a toxin (ccdB) under the cspA promoter is expressed but counteracted by the antitoxin ccdA (<partinfo>BBa_K4244023</partinfo>), which in turn is expressed by being placed after the constitutive LacUV5 (<partinfo>BBa_M36801</partinfo>) [1]. At lower temperature the cspA stabilizes and ccdB overexpresses ccdA, which leads to cell death.  
  
  

Revision as of 09:32, 11 October 2022


placUV5:ccdA + pcspA:ccdB

The pcspA (BBa_K4244021) is a constitutive promoter that has a high rate of transcription but contains a long 5′ UTR of 159 bp that, at 37°C, acquires an unstable secondary structure and leads to its degradation. On the other hand, at lower temperatures it forms a stable configuration, which allows translation. In this case, a toxin (ccdB) under the cspA promoter is expressed but counteracted by the antitoxin ccdA (BBa_K4244023), which in turn is expressed by being placed after the constitutive LacUV5 (BBa_M36801) [1]. At lower temperature the cspA stabilizes and ccdB overexpresses ccdA, which leads to cell death.


Figure 1: The temperature sensitive survival assay shows that the pcspA system with the toxin-antitoxin (ccdA-ccdB) no colonies are present in the plates incubated at temperatures below 37°C compared to the control, pcspA system with reporter gene.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 44
    Illegal PstI site found at 1089
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 1089
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 44
    Illegal PstI site found at 1089
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 44
    Illegal PstI site found at 1089
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 985


References

1.Stirling, F., Bitzan, L., O'Keefe, S., Redfield, E., Oliver, J., Way, J., & Silver, P. A. (2017). Rational Design of Evolutionarily Stable Microbial Kill Switches. Molecular cell, 68(4), 686–697.e3. https://doi.org/10.1016/j.molcel.2017.10.033

Usage and Biology