Difference between revisions of "Part:BBa K4137000"
 
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<partinfo>BBa_K4137000 short</partinfo>
 
<partinfo>BBa_K4137000 short</partinfo>
  
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BBa_K4137000 codes for the CcdA protein, a component of the CcdA-CcdB toxin-antitoxin system. CcdA binds with CcdB which inhibits CcdB from targeting DNA gyrase thus preventing DNA double-strand breaks. A 6xHis tag was added for protein purification and identification.  
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BBa_K4137000 codes for the CcdA protein, a component of the CcdA-CcdB toxin-antitoxin system. CcdA binds with CcdB which inhibits CcdB from targeting DNA gyrase thus preventing DNA double-strand breaks.  
  
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[[Image:ccda-his.png|800px|thumb|center|Fig.1 ccdA with C-Terminal 6x His-Tag.]]
 
[[Image:ccda-his.png|800px|thumb|center|Fig.1 ccdA with C-Terminal 6x His-Tag.]]
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[[File:CAU CrtE Fig1.png|800px|thumb|center|Fig.1 The site-specific mutation of CrtE.]]
 
  
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===Usage and Biology===
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===Characterization===
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We obtained the sequence for ccdA from IDT through gblock synthesis. Since we aimed to develop determine how different combinations of our toxin-antitoxin system components (ccdA, ccdB, mleR) affect one another's activity to verify our kill switch theory, we designed our constructs with biobrick restriction sites that bracket our genetic components--to insert one of our components into our backbone pSB1C3, we would cut at EcoRI and SpeI for both the insert and the vector. We also flanked each component with a purification tag to observe their expression levels.
  
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[[Image:ccda-cut.png|800px|thumb|center|Fig.2 ccdA with E-S cut sites.]]
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===Sequence and Features===
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K4137000 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K4137000 SequenceAndFeatures</partinfo>
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<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  

Latest revision as of 02:54, 10 October 2022


CcdA-6xHis Antitoxin

BBa_K4137000 codes for the CcdA protein, a component of the CcdA-CcdB toxin-antitoxin system. CcdA binds with CcdB which inhibits CcdB from targeting DNA gyrase thus preventing DNA double-strand breaks.


Fig.1 ccdA with C-Terminal 6x His-Tag.

Characterization

We obtained the sequence for ccdA from IDT through gblock synthesis. Since we aimed to develop determine how different combinations of our toxin-antitoxin system components (ccdA, ccdB, mleR) affect one another's activity to verify our kill switch theory, we designed our constructs with biobrick restriction sites that bracket our genetic components--to insert one of our components into our backbone pSB1C3, we would cut at EcoRI and SpeI for both the insert and the vector. We also flanked each component with a purification tag to observe their expression levels.

Fig.2 ccdA with E-S cut sites.

Sequence and Features

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]