Coding

Part:BBa_K762002

Designed by: Menuka Samaranayake   Group: iGEM12_Wisconsin-Madison   (2012-09-24)

TCC Positive Control

Positive control for UW-Madison 2012's Translational Coupling Cassette. Consists of a constitutive promoter (J23102) and a gene encoding a green fluorescent protein (E0040) with the stop codon removed. When inserted upstream of the TCC (K762000), translation will continue and produce a mixture of green and red fluorescence (TCC contains a RFP reporter gene). The GFP is the preferred positive control due to the ease at which its production can be assayed. Other options would be more complicated, including the need to do a Western Blot and purify the His-tagged protein. This construct verifies that the gene of interest is properly translated due to detection of the GFP after the hairpin is broken.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1361
    Illegal AgeI site found at 1473
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 685


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Categories
Parameters
None