Part:BBa_K1956020
RIBOSOFF with plasmid control
RIBOSOFF 1 with plasmid control
Team IIT Madras 2016's Characterization
Trigger RNA: A part of RNA molecule, which codes for lacI protein.
Switch RNA: It was designed to activate the translation process by allowing the ribosomal machinery to bind to RBS, placed upstream of GFP protein coding part. After interacting with the RNA molecules of lacI gene, RIBOSOFF changes it's confirmation to mask the RBS and represses the translation process.
We performed our RIBOSOFF experiment in E. coli DH5alpha. We transformed our cloned device BBa_K1956020, which has RIBOSOFFlacI switch under J23106 constitutive promoter, upstream of GFP coding part and RFP generator under lacI+pL promoter.
We prepared two types of cells: 1. BBa_K1956020 in pSB1A2 + BBa_K1956034 in pSB1K3, 2. BBa_K1956020 only.
We grew the cells in LB broth media. After, 16 hrs of growth, we inoculated secondary cultures and collect samples at 12 hour of growth.
Positive Control: Device K1956015, which produces GFP and RFP proteins.
Negative Control: Device K1956034, which produces lacI mRNA molecules.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1849
Illegal AgeI site found at 1961 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 847
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