Test pLsrA/cI

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:

pBAD→RBS+LsrR+TT

plsrA/cI→GFP does not glow

I13453+B0034+K091001+B0015+K091106+E0240

add AI-2

pBAD→RBS+LsrR+TT

plsrA/cI→GFP glows

I13453+B0034+K091001+B0015+K091106+E0240

LsrR	cI	AI-2 + GFP	AI-2 - GFP
0	0	1	1
1	0	1	0
0	1	0	0
1	1	0	0

Ligation plan for test sequences

K091106 ligated with E0240

need to ligate some promoter (digest with E/S) with an RBS (digest E/X) and ligate lsrR(digest with E/S) with a double terminator (digest with E/X)

then ligate the promoter+RBS to the 5' end of the lsrR+TT parts together

then ligate that part 5' to the plsrA/cI+GFP