Part:BBa_K1210003

PK401 tagged dual coding test construct

Reporter construct for measuring the frameshifting efficiency of the PK401 pseudoknot. Includes a His-tagged CFP before the PK401 pseudoknot, and two different polypeptides in the 0 and -1 reading frames with different protein tags.

If translation is halted at the pseudoknot due to the inability to melt the mRNA secondary structure, a 27.7 kDa protein is produced of just CFP-His. In the 0 frame if read-through of the pseudoknot occurs, a fragment of the IAPV dual coded gene along with a strep-tag II will be translated creating a 33.3 kDa protein of CFP-His-PK401-IAPV-StrepII. In the -1 frame if read-through of the pseudoknot occurs, a fragment containing the second dual coded IAPV gene along with a GST tag is produced. This contains CFP-His-PK401-IAPV2-GST, a 60.9 kDa protein.

Sequence and Features

Assembly Compatibility:

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

  Illegal BamHI site found at 1457
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 90
  Illegal BsaI.rc site found at 1600