Reporter

Part:BBa_K1758101

Designed by: Team Bielefeld-CeBiTec 2015   Group: iGEM15_Bielefeld-CeBiTec   (2015-08-17)
Revision as of 19:20, 19 September 2015 by Mrfreeze (Talk | contribs)

Translation enhancing 5-UTR + sfGFP

This part includes a our designed, translation enhancing 5'-untranslated region (5'-UTR; BBa_K1758100), in front of sfGFP coding sequence (BBa_I746916). It also contains a double terminator made of B0010 and B0012. The translation enhancing sequence improves the binding of the mRNA to the ribosome ([http://www.ncbi.nlm.nih.gov/pubmed/2676996 Olins et al. 1989]) and is especially helpful in in vitro cell free protein synthesis. The sequence contains a spacer of 10 adenine bases which has been shown to further enhance translation efficiency ([http://www.ncbi.nlm.nih.gov/pubmed/23927491 Takahashi et al. 2013]).


Usage and Biology

By adding a promoter of choice, sfGFP is produced. The high efficiency of our designed 5'-UTR ensures reporter production even when weak promoters are employed.

Characterization

This part has been used in a variety of experiments, including experiments with BBa_K1758102, BBa_K1758377, BBa_K1758312, BBa_K1758314, BBa_K1758323, BBa_K1758325, BBa_K1758332 and others. For details see the corresponding part or our wiki. The sequence was in general cloned via Gibson assembly.


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
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 59


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Categories
Parameters
None