Difference between revisions of "Part:BBa K4137000"
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===Characterization=== | ===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 cut at EcoRI and SpeI for both the insert and the vector. We flanked each component with a purification tag to observe their expression levels. | 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 cut at EcoRI and SpeI for both the insert and the vector. We flanked each component with a purification tag to observe their expression levels. | ||
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| + | [[Image:ccda-cut.png|800px|thumb|center|Fig.1 ccdA with C-Terminal 6x His-Tag.]] | ||
===Sequence and Features=== | ===Sequence and Features=== | ||
Revision as of 02:47, 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.
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 cut at EcoRI and SpeI for both the insert and the vector. We flanked each component with a purification tag to observe their expression levels.
Sequence and Features
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]