Part:BBa_K3802000

PgsA-MlrA fusion protein

Sequence and Features

This fusion protein aims to anchor MlrA, or microcystinase, an enzyme capable of degrading freshwater toxin microcystin-LR (MC-LR) to the outer membrane of E. coli. This part puts together a codon-optimized subunit of PgsA (poly-γ-glutamic acid synthetase) (BBa_K2963020), an anchoring motif to MlrA with 6x His (BBa_K1907002), with a 2 amino acid Gly-Ser linker (BBa_J18920). The addition of the His tag to MlrA gives this part the ability to be purified using Nickel affinity chromatography and detected using Western blotting with anti-His antibodies. This BioBrick was put under the control of an IPTG inducible ptac promoter (BBa_K3802001), as using a constitutive promoter could potentially lead to the depletion of resources for the bacteria.

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 1792
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 827
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 813
  Illegal NgoMIV site found at 2323
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 304
  Illegal BsaI.rc site found at 2160