Part:BBa_I759015:Design
cis3-repressed, tet-regulated YFP
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 981
Illegal NheI site found at 1004 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
This part is the YFP sequence down-regulated by the cis repressive element. The Ptet promoter and TetR gene, which is under the control of a constitutive promoter, establish an inducible system in which transcription can be activated by the addition of anhydrotetracycline (aTc).
Cis3 is a modified version of construct “CR12,” which has been shown to repress transcripts by Collins et. al. Modifications involve a change in the number of bulges and end-loop size which affect the free energy of interaction within the cis structure. The key principal in the design was to find a structure that offered tight repression of the gene of interest but also allowed for activation upon addition of a trans-activating element. A lower free energy indicates tighter binding and a decreased favorability for forming a compliment with the trans element. The cis3 construct was designed to be activated by trans1 and trans2. Design specifics: free energy = -11.7kcal; 9bp loop, 4 bulges
Source
Caltech iGEM 2007. The cr element is a synthetic oligo which has been inserted into part I759009.
References
Reference: Isaacs FJ, Dwyer DJ, Ding C, Pervouchine DD, Cantor CR, and Collins JJ. Engineered riboregulators enable post-transcriptional control of gene expression. Nat Biotechnol 2004 Jul; 22(7) 841-7.