Part:BBa_K608407

Precipitator fused with GST tag

Protein domain of Precipitator N-terminally fused to a GST tag domain. The GST-tag has the size of 220 amino acids (roughly 26 KDa). It is fused to the N-terminus of the precipitator protein BBa_K608406 and includes a PreScission Protease domain for cleavage of the GST tag during protein purification.

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Artificial Leucine Rich Repeat(LRR) with C and N-terminal hagfish domain fragments capping the artifical middle part. This part is one version of three different, designed to bind nickel by histidines which are grouped together pointing away from the horseshoe shaped protein. Bacterial LRR Consensus of the central LRR fragment: LxxLxLxxNxLxxLPxxLPxx Protein code: CPSRCSCSGTEIRCNSKGLTSVPTGIPSS ATRLELESNKLQSLPHGVFDK LTQLTKSNNHLHSLPDNLPAS LEVLDVSNNHLHSLPDNLPAS LEVLDVSNNHLHSLPDNLPAS LEVLDVSNNHLHSLPDNLPAS LEVLDVSNNHLHSLPDNLPAS LEVLDVSNNHLHSLPDNLPAS LKELALDTNQLKSVPDGIFDR LTSLQKIWLHTNPWDCSCPRIDY LSRWLNKNSQKEQGSAKCSGSGKPVRSIICP

This protein can be used to complex Nickel or Cobalt. The principal mechanism is comparable to Ni-NTA columns, as chelates the ions. Free binding sites of the ions are then exposed, so that a His-tagged protein can attach to them. The design of the protein is of a particular interest, too. LRR are highly conserved motifs throughout evolution. They appear in all kingdoms of life in almost every thinkable role (Ligases, Receptors, Toxins etc.). Their core is highly conserved and provides a very stable backbone, while the intermediate, non-conserved aminoacids are almost freely interchangeable.

This protein can be used to complex up to 4 Nickel or Cobalt. However the principal design oft he protein is of a particular interest, too. LRR are highly conserved motifs throughout evolution. They appear in all kingdoms of life in almost every thinkable role (Ligases, Receptors, Toxins etc.). Their core is highly conserved and provides a very stable backbone, while the intermediate, non-conserved aminoacids are almost freely interchangeable. We only submitted one of the three versions, to reduce redundancy in the registry. Please contact us for any questions.

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 958
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal XhoI site found at 784
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 1416
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 85