Device

Part:BBa_K1210000

Designed by: Zak Stinson   Group: iGEM13_Lethbridge   (2013-08-02)

LacI-CFP-PK401-YFP

Reporter construct for measuring the frameshifting efficiency of the PK401 pseudoknot. Produces a reporter of framshift-induction. If translation continues in the 0 frame, a stop codon will be reached to give CFP as the final product. If frameshifting occurs due to the PK401 pseudoknot, translation will continue in the -1 frame to produce a CFP-YFP fusion protein with which FRET can be measured.

This construct was designed to verify the applicability of frameshift inducing elements in the programming of protein expression ratios.

Datasheet for Part BBa_K1210000 in E. coli strain DH5α.
ULeth2013iGEM-PK401 data sheet2.jpg

notes:

"TAATAA" stop codons were removed from the end of eCFP

"TT" two nucleotide insertion was added after the pseudoknot to put eYFP into the -1 frame.

Reference

(1) Hansen T.M., Reihani S.N., Oddershede L.B., Sørensen M.A. (2007)Correlation between mechanical strength of messenger RNA pseudoknots and ribosomal frameshifting. Proc Natl Acad Sci 104(14), 5830-5.

Sequence and Features

Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None