Part:BBa_K2120311
B0015+tetR+B0034+pBAD-AraC+B0015+ptet+B0034+RFP+B0015
pBAD is a promoter induced by arabinose and arac is the repressor. We use it to test the expression of reporter gene in our validation circuit .And tetR is an inhibitor which is built to repress the expression of ptet promoter. We use it to control the “on” or ”off” state of toxin gene. RFP is the reporter gene. In order to test these do not interfere with each other , We reserve BBa_K2120303 .
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 911
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1076
Illegal AgeI site found at 2839
Illegal AgeI site found at 2951 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 1093
Functional Parameters
P-Slackiller is the iGEM 2016 project of BIT-China. In industrial produce, plasmid plays an important role in produce because it carries function gene which codes something we wanted. But bacteria will lose the functional plasmid during the division then become a “Slacker” decreasing the efficiency. In order to kill the “Slacker”, we aim to design logical gene switch to sense the copy number variety.
However, it is difficult to control the plasmid copy number. So we choose BBa_K808000 whose expression level of downstream gene can vary continuously for imitating the change of plasmid copy numbers. And connect the logical switch downstream.
The logical switch we select the relationship of inhibitor-promoter. In our building work, we finally decide two kinds of inhibitor-promoter, cI-pR and tetR-pTet. Then we add RFP in the downstream of promoter as a reporter for indicating “on” or “off”.
First, we built K2120309. But during our test, we found the expression of araC would influence the expression of RFP. So we added another B0015 between araC and pR.
Beside, we also have built a similar device, BBa_K2120311, using tetR-pTet. But we had no time to test it.
| None |