Difference between revisions of "Part:BBa K4137011"

Revision as of 05:30, 10 October 2022

CcdB coding sequence + 6xHis

The above sequence encodes for the CcdB protein, a component of the CcdA-CcdB toxin-antitoxin system. CcdB binds with CcdA which inhibits CcdB from targeting DNA gyrase thus preventing DNA double-strand breaks. The 6xHis tag is used for protein purification and identification.

Fig.1 ccdB with N terminal 6x his tag.

Characterization

We obtained the sequence for ccdB 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.

Sequence and Features