Part:BBa_K4137001

CcdB-Myc Toxin

BBa_K4137001 codes for the CcdB protein, a toxin to many bacteria including E. coli and B. subtilis. Through the formation of a CcdA-CcdB complex by binding with CcdA, CcdB loses its ability to interact with DNA gyrase thus losing its toxicity. A Myc tag is added for protein purification, identification, and distinction from CcdA.

Fig.1 ccdB with C-Terminal Myc-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.

Fig.2 ccdB 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
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 216