- 10COMPATIBLE WITH RFC
- 12Illegal NheI site found at 3206
- 21Illegal BamHI site found at 3145
- 23COMPATIBLE WITH RFC
- 25Illegal AgeI site found at 2980
- 1000Illegal SapI site found at 2962
PETase-CBD-HlyA was designed as a G block and obtained from Integrated DNA Technologies. Sequence for isPETase was obtained from Joo et al. 2018 and codon optimized for E. Coli K12 Sequence for CBD was obtained from BBa_K1321014 from Imperial in 2014. A NC linker was added to the sequence with an HlyA secretion tag. None of these existing parts were assembled by 3a assembly since the scar site contains a stop codon, which would not allow us to assemble a modular fusion protein. HlyBD was obtained from BBa K1166002.
This part is equipped with two promoters each individually activating a region of the protein. The T7 promoter controls the expression of PETase-CBD-HlyA whereas an arabinose promoter controls the expression of the secretion system HlyBD.
BBa_I712024 BBa_B0034 BBa_K132104 BBa_K1166002
Joo, S. et. al. Structural insight into molecular mechanism of poly(ethylene terephthalate) degradation. Nature Communications 9 (2018)