Collection list
Here is the list of our part collection:
| Part Number | Name | Description |
| BBa_K2036009 | pRM-GFP-LVAssrAtag | A control group of Cro and pRM interaction characterization |
| BBa_K2036010 | Cro-TT-pRM-RBS-GFP-LVAssrAtag | To test Cro and pRM interaction intensity. |
| BBa_K2036011 | pRE-GFP-LVAssrAtag | A control group of CII and pRE interaction |
| BBa_K2036013 | RBS-CII-TT-pRE-RBS-GFP-LVAssrAtag | To characterize CII’s interaction with pRE. |
| BBa_K2036014 | RBS-CIII-RBS-CIII-RBS-CII-TT-pRE-RBS-GFP-LVAssrAtag | To characterize CIII’s interaction with Ftsh. |
| BBa_K2036015 | RBS-CII-RBS-CII-RBS-CII-TT-pRE-RBS-GFP-LVAssrAtag | Tandem of CII amplifies its function. |
| BBa_K2036016 | pR-GFP-LVAssrAtag | A control group of CI and pR interaction. |
| BBa_K2036017 | CI-TT-pR-RBS-GFP-LVAssrAtag | To characterize CI and pR interaction. |
| BBa_K2036027 | pRE-RBS-Cro-RBS-CII-TT-ptrp-RBS-CI-TT-pR-RBS-CIII-RBS-RFP-LAAssrAtag-TT-pRM-RBS-GFP-LVAssrAtag | To verify the function of the whole circuit with T7 and ptrp sensing the signals. |
| BBa_K2036028 | placm-pRE-RBS-Cro-RBS-CII-TT-patp2-RBS-CI-TT-pR-RBS-CIII-RBS-ompA-iLDH-TT-pRM-RBS-beta-galactosidase | To verify bi-stable function of eukaryotic version of signal filter. |
The part collection well documents the prokaryotic regulatory circuit of our project with every detailed characterization of transcriptional related proteins and promoter.
The whole circuit employs bacteriaphage lambda operon and is redesigned as a positive feedback tri-stable switch. Users can define two inputs and three kinds of outputs to make a sense-response system or an analysis-report system.
To characterize the internal protein-protein and protein-promoter reactions, we build four sets of test circuits:
1.Cro and pRM inhibition test:
| Part Number | Name | Description |
| BBa_K2036020 | RBS-CI-TT-pR-RBS-CIII-RBS-RFP-LAAssrAtag-TT-pRM-RBS-GFP-LVAssrAtag | To construct the verification circuit. |
| BBa_K2036027 | pRE-RBS-Cro-RBS-CII-TT-ptrp-RBS-CI-TT-pR-RBS-CIII-RBS-RFP-LAAssrAtag-TT-pRM-RBS-GFP-LVAssrAtag | To verify the function of the whole circuit with T7 and ptrp sensing the signals. |
In ideal conditions, RFP is supposed to be visualized through fluorescence microscope when induced by IPTG. And after adding IAA into the medium, our host strain will turn to the other state of GFP expression. And because of LVAssrA tag’s effect, the RFP to GFP expression transition will be relatively obvious.
Application cicuit
To provide an application example, we constructed an application circuit based on this version to help relief lactose intolerance (more details see to application page) .
| Part Number | Name | Description |
| BBa_K2036028 | placm-pRE-RBS-Cro-RBS-CII-TT-patp2-RBS-CI-TT-pR-RBS-CIII-RBS-ompA-iLDH-TT-pRM-RBS-beta-galactosidase | To verify bi-stable function of eukaryotic version of signal filter. |
In the bi-stable switch of prokaryotes, we set the gene of interest 1 as iLDH and gene of interest 2 as beta-galactosidase. For sensing the signal, we choose lactose inducible promoter plac(mutant) and base inducible promoter patp2 (works when the pH of microenvironment is about 7~9).
And we hope more team to join and add more posibilities to the circuit!
