Composite

Part:BBa_K5117045

Designed by: Jenny Sauermann, Lilli Kratzer, Katrin Lehmann   Group: iGEM24_TU-Dresden   (2024-09-28)
Revision as of 16:33, 1 October 2024 by Jesa98 (Talk | contribs)


PcotYZ-BsRBS-BhrPET-L2-CotY-B0014

This part serves as transcriptional unit composed of:

  • promoter PcotYZ of Bacillus subtilis (BBa_K5117021)
  • ribosome binding site of Bacillus subtilis (BBa_K5117000)
  • gene of the uncultured bacterium HR29 encoding a polyethylene terephthalate hydrolase (PETase, EC 3.1.1.101),
    • a homologue of the leaf-branch compost cutinase (LCC), codon optimized for Bacillus subtilis (Xi et al. 2021) without any signal peptide,
    addition of a long flexible linker (L2) downstream of the coding sequence encoding the amino acids (GGGGS)4 (BBa_K5117033)
  • cotY gene of Bacillus subtilis (BBa_K5117022)
  • bidirectional terminator B0014 (BBa_B0014)


Biosafety level: S1

Target organism: Bacillus subtilis

Main purpose of use: Immobilization of BhrPET on the spore crust of B. subtilis (spore surface display)

Application: Degradation of PET


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
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1063
    Illegal AgeI site found at 695
    Illegal AgeI site found at 740
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None