Composite

Part:BBa_K2787013

Designed by: Haoer Shi   Group: iGEM18_ShanghaiTech   (2018-10-09)


GFP under B8 orthogonal RBS with orthogonal 16S rRNA BBa_K2787041

The 16S rRNA BBa_K2787040 under constitutive promoter BBa_J23100(BBa_K2787008) and pCons-oRBSB8-GFP(BBa_K2787001) are linked into one plasmid to quantify the orthogonality of the TJU and B8 ribosome system.

Usage and Biology

The plasmid with GFP under C9 orthogonal RBS with orthogonal 16S rRNA A2 is constructed.

Shanghaitech2018-oribo-orbs-07-07.png
ShanghaiTech2018-b8 rbs.png

The GFP under control of B8 orthogonal RBS is tested within different 16s rRNAs. The fluorescence with C9 orthogonal ribosome is higher than host ribosomes and A2,TJU ribosomes, which means the orthogonality between that of C9 ribosome is not as expected.

ShanghaiTech-TJU 16S.png

The GFP under control of different orthogonal RBS is tested within TJU orthogonal 16s rRNAs. There is no fluorescence difference between each group, means that the A2 ribosomes is orthometric to other RBSs.

A2-B8-C9-Orthogonality.png

The expression of the gene of interest is tested between different orthogonal RBS and different orthogonal ribosomes. 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
    Illegal NheI site found at 798
    Illegal NheI site found at 821
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1661
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1675
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 707


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