Composite

Part:BBa_K4137007

Designed by: Andrea Huang   Group: iGEM22_GEMS_Taiwan   (2022-08-19)


pLac+RBS+CcdB-Myc+B1006

This part produces a constant expression of ccdB toxin under the LB medium, inhibiting the GyrA subunit of DNA gyrase and prevents negative supercoiling of DNA. An accumulation of ccdB without sufficient suppression by antitoxin ccdA leads to risks in host viability.

Fig.1 Construct design for ccdB toxin secretion.

Construct Design

We attached a Myc epitope Tag downstream of the ccdB sequence for purification purposes. A pLac promoter is attached upstream to the side of the open reading frame (ccdB-Myc coding complex) and RBS (AAGGAG). The terminator BBa_B1006 is attached downstream of the sequence.


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 271


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