Difference between revisions of "Part:BBa K4367014"

 
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<partinfo>BBa_K4367014 short</partinfo> <br>
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ABI-cTEV is a part of the ABI/PYL-splitTEV system, which is used as a control for other experiments. Essentially, the complementation of split TEV can be induced with the addition of abscisic acid to the system.
  
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<h2>Description</h2>
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===Usage and Biology===
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The design of ABI-cTEV is essentially identical to what was used in this reference [1], but with the addition of a 6xHis-tag and Part-3-Overhangs for Modular Cloning. PYL and ABI can dimerize if abscisic acid is present [1]. When PYL-nTEV dimerises with ABI-cTEV, the split TEV can complement each other and regain enzymatic function.
  
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<span class='h3bb'>Sequence and Features</span>
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<h2>Usage</h2>
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It was planned to use the ABI/PYL-splitTEV system as a control for the dCas9-splitTEV system, and to use as stepping stone for troubleshooting what parts could be dysfunctional. For example, if the ABI/PYL system works but not the dCas9-splitTEV system, it's shown that the testing conditions can allow for complementation of split TEV and the problem is perhaps because of the dCas9 or gRNA.
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<h2>Future design considerations</h2>  
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If for some reason the ABI/PYL system would not be desirable, there is an alternative: The proteins FKBP and FRB, which uses the chemical rapamycin to induce their dimerisation [1].
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<h2>Sequence and Features</h2>  
 
<partinfo>BBa_K4367014 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K4367014 SequenceAndFeatures</partinfo>
  
  
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<h2>References</h2>  
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===Functional Parameters===
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<partinfo>BBa_K4367014 parameters</partinfo>
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[1] Tina Fink, Jan Lonzarić, Arne Praznik, Tjaša Plaper, Estera Merljak, Katja Leben, Nina Jerala, Tina Lebar, Žiga Strmšek, Fabio Lapenta, Mojca Benčina & Roman Jerala (2019). Design of fast proteolysis-based signaling and logic circuits in mammalian cells. Available at: https://www.nature.com/articles/s41589-018-0181-6
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Revision as of 21:31, 11 October 2022


ABI-cTEV
ABI-cTEV is a part of the ABI/PYL-splitTEV system, which is used as a control for other experiments. Essentially, the complementation of split TEV can be induced with the addition of abscisic acid to the system.


Description

The design of ABI-cTEV is essentially identical to what was used in this reference [1], but with the addition of a 6xHis-tag and Part-3-Overhangs for Modular Cloning. PYL and ABI can dimerize if abscisic acid is present [1]. When PYL-nTEV dimerises with ABI-cTEV, the split TEV can complement each other and regain enzymatic function.


Usage

It was planned to use the ABI/PYL-splitTEV system as a control for the dCas9-splitTEV system, and to use as stepping stone for troubleshooting what parts could be dysfunctional. For example, if the ABI/PYL system works but not the dCas9-splitTEV system, it's shown that the testing conditions can allow for complementation of split TEV and the problem is perhaps because of the dCas9 or gRNA.


Future design considerations

If for some reason the ABI/PYL system would not be desirable, there is an alternative: The proteins FKBP and FRB, which uses the chemical rapamycin to induce their dimerisation [1].


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 952
    Illegal BamHI site found at 1034
    Illegal BamHI site found at 1363
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 14
    Illegal BsaI.rc site found at 1370
    Illegal SapI.rc site found at 1270


References

[1] Tina Fink, Jan Lonzarić, Arne Praznik, Tjaša Plaper, Estera Merljak, Katja Leben, Nina Jerala, Tina Lebar, Žiga Strmšek, Fabio Lapenta, Mojca Benčina & Roman Jerala (2019). Design of fast proteolysis-based signaling and logic circuits in mammalian cells. Available at: https://www.nature.com/articles/s41589-018-0181-6