Difference between revisions of "Part:BBa K4137000"

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

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]

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K4137000 short</partinfo>
-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.
+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.
 [[Image:ccda-his.png|800px|thumb|center|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.
+[[Image:ccda-cut.png|800px|thumb|center|Fig.2 ccdA with E-S cut sites.]]
 ===Sequence and Features===
+<span class='h3bb'>Sequence and Features</span>
 <partinfo>BBa_K4137000 SequenceAndFeatures</partinfo>