Part:BBa_K2036010
Cro-TT-pRM-RBS-GFP-LVAssrAtag
It is a negtive control of GFP expression. When inserted into a expression plasmid downstream an inducible promoter, it will form a NO gate. Take PETDuet-1 as an example, after constructing Cro-TT-pRM-RBS-GFP-LVAssrAtag-PETDuet-1, IPTG can trigger T7 promoter in the plasmid, and Cro can bind to a certain site within pRM, so that GFP will not be produced. And we get a NO gate of GFP generator.
HUST-China 2016 build this circuit to test Cro and pRM interaction intensity with contol group: pRM-GFP-LVAssrAtag (BBa_K2036009)
Protein&promoter
--Cro and pRM
Preliminary experiments of LVAssrAtag
In order to prove that our toolkit is efficient to switch two interest genes’ expression from GFP to RFP and to eliminate the accumulation of expressed protein to interfere our measurement. We fused a degradation tag at the amino terminal of our reporter. And we used plac from the Registry(BBa_J04500) to characterize the degradation tag LVAssrA. We use IPTG with final concentration of 1mM to induce the GFP-LVAssrAtag and measure the relative fluorescence through plate reader with excitation wavelength 495nm.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1036
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