Test pLsrA/cI promoter
The pLsrA/cI promoter is constitutively off and is repressed by the lsrR protein. The addition of AI-2 should de-repress the lsrR operon causing it to glow. The addition of the cI protein should repress the promoter, causing it to not glow.
Test pLsrA/cI repression:
plsrA/cI→RBS+GFP+TT does not glow
|LsrR||cI||AI-2 + GFP||AI-2 - GFP|
Ligation plan for test sequences
Digest RBS with S/P, lsrK with X/P; do ligation.
Digest RBS with S/P, lsrR with X/P; do ligation.
Digest pBAD with S/P, RBS+lsrK with X/P; do ligation.
Digest TT with R/X, RBS+lsrR with R/S; do ligation.
Digest RBS+lsrR+TT with R/X, pBAD+RBS+lsrK with R/S,; do ligation.
Digest plsrA/cI with S/P, E0240 with X/P; do ligation.