Composite

Part:BBa_K2905018

Designed by: David Viguilla   Group: iGEM19_Alma   (2019-10-16)
Revision as of 14:06, 21 October 2019 by Djcamenares (Talk | contribs)


TAG-Read through reporter

A reporter construct - normally expresses a blue chromoprotein, but will also produce a red protein if the TAG codon does not terminate translation

Measuring fluorescence and expression: Alma 2019

To determine if the part is working correctly, we transformed it into DH5a, MRA8 (A temperature sensitive RF1 strain), and C321.ΔA.exp (a strain in which RF1 has been deleted - C321.ΔA.exp was a gift from George Church; Addgene plasmid # 49018). The expectation is that DH5a should display a blue color, while MRA8 and C321 may show a higher degree of red. C321 grew slowly, but displayed results similar to that of MRA8 - it is not included on the representative plate below.

T--Alma--readthrough reporter1.png

This plate clearly shows that the reporter is functional for producing the blue chromoprotein. Visualizing red fluorescence is not as easy, although there seems to be more red hue in MRA8 than in DH5a. Measuring fluorescence is also complicated by the fact that the chromoprotein, cjBlue, and RFP share absorption spectra.

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
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1525
    Illegal AgeI site found at 1637
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
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