Constitutive Promoter

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Presentation transcript:

Constitutive Promoter Key for Diagrams “AND” Gate- both inputs are necessary for an output. Constitutive Promoter Inducible Promoter Repressible Promoter Ribosome Binding Site Open Reading Frame Terminator

Device 1- Oxidative Stress Induced RA Generator NO Input 8 3 4 2 SoxS Promoter BC-RA Converter Generator Retinoic Acid Const. SoxR Generator 5 Metabolite 7 Beta-Carotene 6 Const. BC Generator

Crt Cluster- BC Producer Device 1- Oxidative Stress Induced RA Generator SoxR NO SoxR gene SoxS Promoter Blh- BC to Retinal RALDH II- Retinal to RA NO Sensor BC-RA Converter Retinoic Acid Crt Cluster- BC Producer Beta-Carotene Generator

Device 1- Oxidative Stress Induced RA Generator NO SoxR NO Sensor BC to RA Converter J23100 Const Prom. B0034 RBS k2230000 SoxR gene B0015 TT k2230001 SoxS Prom. B0034 RBS k223035 Blh gene B0034 RBS k223036 Raldh II gene B0015 TT BC to Retinal Retinal to RA Beta-Carotene Generator Retinoic Acid J23100 Const Prom. B0034 RBS ? Crt Cluster B0015 TT

Device 2- Il-6 Secretion System Il-6 Generator Constitutive Export System Generator POPS Invertor 5 MT 5 4 3 2 1

Repressible 5MT Operator Device 2 5MT Level Sensitive Sensor Il-6 Generator Repressible 5MT Operator Il-6 gene HlyA Linker Il-6 Secreted Export System TolC gene Hly R,B,D,C

Device 2 5MT Level Sensitive Il-6 Generator Sensor Il-6 Secreted k223023 Trp Operator B0034 RBS B0015 TT k223022 Il-6 + HlyA linker Allows Il-6 to leave through the export channel Il-6 Secreted Export System J23100 Const Prom. B0034 RBS k223025 Hly R,B,D,C B0015 TT k223024 TolC gene Create an Export Channel and Chaperone

Fourth Round of Cloning Device 2 Fourth Round of Cloning 2.4a Trp Operator, RBS,Il-6+HlyA,TT + Rbs, TrpR, TT (Cloned from step 2.3a) + (Cloned from step 2.1e) + J23100 + #? (Const. Promoter) (Synthesized de novo as 1part = RBS + trpR + TT) #k223023 + B0034 + k223022 + B0015 (Trp operator-) (RBS) (Il-6 and HlyAs linker) (TT) synthesized) Cut w/ EcoR1/Spe1 Cut w/ EcoR1/Xbal

Device 2 + Fourth Round of Cloning 2.4b Trp Operator + Rbs, TolC, HlyR,B,D,C, TT (will be registered) + (Cloned from step 2.3b) + #k223023 (Trp operator - synthesized) B0034 + k223024 + k223025 + B0015 (RBS) (TolC ) (HlyR,B,D,C) (TT) Cut w/ ? Cut w/ ?