Difference between revisions of "Part:BBa M50499"

Revision as of 23:43, 11 December 2018

Catabolite Activator Protein (CAP) Repressed Promoter

We placed the constitutive promoter (BBa_S05450, iGEM) immediately upstream from a catabolite activator protein (CAP) binding site (BBa_M36547, iGEM) in Plasmid E.

For our experiments, we placed this part upstream from GFP in order to assay expression levels through fluorescent intensity. This construct is an adaption of the natural lac operon, which is positively regulated by CAP, since the CAP binding site is naturally upstream of the RNA polymerase binding site. In this construct, we put the CAP binding site downstream in order to repress transcription when CAP binds due to steric hindrance.

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal XbaI site found at 1059
Illegal PstI site found at 688
Illegal PstI site found at 849
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 144
Illegal NheI site found at 167
Illegal PstI site found at 688
Illegal PstI site found at 849
21
COMPATIBLE WITH RFC[21]
23
INCOMPATIBLE WITH RFC[23]
Illegal XbaI site found at 1059
Illegal PstI site found at 688
Illegal PstI site found at 849
25
INCOMPATIBLE WITH RFC[25]
Illegal XbaI site found at 1059
Illegal PstI site found at 688
Illegal PstI site found at 849
1000
COMPATIBLE WITH RFC[1000]

@@ Line 3: / Line 3: @@
 <partinfo>BBa_M50499 short</partinfo>
-We placed the constitutive promoter (BBa_S05450, iGEM) immediately upstream from a CAP binding site (BBa_M36547, iGEM) in Plasmid E.
+We placed the constitutive promoter (BBa_S05450, iGEM) immediately upstream from a catabolite activator protein (CAP) binding site (BBa_M36547, iGEM) in Plasmid E.
-For our experiments, we placed this part upstream from GFP in order to assay expression levels through fluorescent intensity.
+For our experiments, we placed this part upstream from GFP in order to assay expression levels through fluorescent intensity. This construct is an adaption of the natural lac operon, which is positively regulated by CAP, since the CAP binding site is naturally upstream of the RNA polymerase binding site. In this construct, we put the CAP binding site downstream in order to repress transcription when CAP binds due to steric hindrance.
 <!-- Add more about the biology of this part here
 ===Usage and Biology===
+Since CAP binding is inversely correlated to glucose concentrations (and positively correlated with cyclic AMP (cAMP) levels), this promoter exhibits glucose-inducibility and cAMP-repressibility. We employed this construct as a potential DNA-based glucose biosensor in E coli.
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