Part:BBa_K1152007

Helper construct for NRP-Indigoidine-tagging

This part serves as a template for the labeling of non-ribosomal peptides with indigoidine. The desired order of modules can be used to replace the suicide gen ccdB. Therefore, selection of positive transformed clones gets a lot easier and nascent non-ribosomal peptides are tagged by the Indigoidine residue attached during synthesis. Since the blue pigment Indigoidine can be seen without auxillary means, analytical procedures can be faciliated a lot. Instead of complex labeling, tagging or work up procedures, simple devices, like a TLC can be used to assess qualitativ differences.

To verify the genotype of our construct colony-PCRs, and restriction digests with Pst1 and EcoRI were conducted. Assembly overhang regions were sequenced as well.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 3061
  Illegal BamHI site found at 891
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 341
  Illegal SapI.rc site found at 4324