Part:BBa_K4623013

Nucleotide Silinker(NS)

Usage and Biology

The Nucleotide Silinker (NS) is a novel engineered protein that covalently links nucleic acid ends to biotinylated adapters. The sequence in FASTA format includes an added HIS tag, enabling purification of the NS protein using a nickel column. Upstream of the mSA sequence, a TrxA (BBa_K3619001) fusion tag is added to aid protein folding and reduce the formation of inclusion bodies in bacterial cells. Following protein expression, cleavage by thrombin exposes the mSA (BBa_K4623001) site, allowing binding to biotinylated functional proteins. The TrwC protein can covalently link specific 5' single-stranded nucleotide sequences, facilitating the conversion between nucleic acids and protein connectors.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 1024
  Illegal PstI site found at 1342
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 1024
  Illegal PstI site found at 1342
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 1024
  Illegal PstI site found at 1342
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 1024
  Illegal PstI site found at 1342
  Illegal AgeI site found at 448
  Illegal AgeI site found at 508
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1147
  Illegal SapI.rc site found at 862