Device

Part:BBa_K1210003

Designed by: Harland Brandon   Group: iGEM13_Lethbridge   (2013-10-15)


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


[edit]
Categories
Parameters
None