Coding

Part:BBa_K4137010

Designed by: YA-JHIH KO   Group: iGEM22_GEMS_Taiwan   (2022-10-07)


CcdA coding sequence + Myc Tag

The above sequence encodes for the CcdA protein, a antitoxin formany bacteria including Escherichia coli. By forming CcdA-CcdB complex by binding with CcdB, CcdB loses its ability to interact with DNA gyrase thus losing its toxicity. The Myc tag is a polypeptide protein tag added for protein purification and identification, which serves similar functions as the 6xHis tag.

Figure 1. ccdA with N-Terminal Myc tag.

Construct Design

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.

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]


[edit]
Categories
Parameters
None