Part:BBa_K3076803:Design

Expression of CgMT driven by T7 promoter under LacO regulation

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal XbaI site found at 47
  Illegal PstI site found at 356
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 356
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal XbaI site found at 47
  Illegal PstI site found at 356
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal XbaI site found at 47
  Illegal PstI site found at 356
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 137
  

Design Notes

The CgMT CDS was synthesized into pET151 expression vector by the ThermoFisher GeneArt platform. The vector contains a 6xHis tag at C terminal of the protein, so the start codon of (BBa K3076100)K3076100 was removed in order to be put into this vector.

Source

The T7 promoter, lacO, RBS, 6xHis tag, and T7 terminator were originally from the pET151 vector. The sequences were obtained from addgene.org. The CgMT sequence was identified by a literature review [1] and extracted from NCBI (ACCESSION: KJ638906). Then, codon-optimized for E. coli by the ThermoFisher GeneArt platform.

References

[1] Jafarian, V., & Ghaffari, F. (2017). A unique metallothionein-engineered in Escherichia coli for biosorption of lead, zinc, and cadmium; absorption or adsorption? Microbiology, 86(1), 73–81. doi: 10.1134/s0026261717010064