IGEM:Stanford/2009/Project Homeostasis/CloningPlan

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Contents

Working Construction Plan Documents

PDF of Construction Overview... Latest version










Device 1 Cloning Plan with Transposed Suffix

Device 1 Cloning Plan with Transposed Suffix

Cloning Step Basic Parts Detailed Parts Antibiotics Enzymes Vector & Size (kb) Insert & Size (kb) Current Cloner Picture
Step 1.1a SoxR/S Promoter + GFP tag RBS-SoxR-TT-SoxS Promoter + RBS-GFP-TT Ant. enzymes Vector ----Insert---- Cloner 1.1a
Progress Notes
Step 1.1b SoxR/S Promoter + Blh RBS-SoxR-TT-SoxS Promoter + RBS-Blh Ant. enzymes Vector Insert Cloner 1.1b
Progress Notes
Step 1.1c Blh + Terminator RBS-Blh + TT Ant. enzymes Vector Insert Cloner 1.1c
Progress Notes
Step 1.1d Constitutive Promoter + Blh (1.1c) Promoter +RBS-Blh-TT Ant. enzymes Vector Insert Cloner 1.1d
Progress Notes
Step 1.1e Constitutive Promoter + Crt Cluster Promoter + RBS-Crt Ant. enzymes Vector Insert Cloner 1.1e
Progress Notes
Step 1.2a Constitutive Promoter + SoxR/s w/ GFP (1.1a) Promoter+ RBS-SoxR-TT-SoxSP-RBS-GFP-TT Ant. enzymes Vector| Insert Cloner 1.2a
Progress Notes
Step 1.2b SoxR/S-Blh (1.1b) + RALDH II RBS-SoxR-TT-SoxSP-RBS-Blh + RBS-RALDH II-TT Ant. enzymes Vector Insert Cloner 1.2b
Progress Notes
Step 1.3a Constitutive Promoter + SoxR/S w/ Blh and RaldhII Promoter+ RBS-SoxR-TT-SoxSP-RBS-Blh-RBS-RALDH II-TT Ant. enzymes Vector Insert Cloner 1.3a
Progress Notes
Controls W/ PQ (+) Constitutive Promoter + GFP tag Promoter+ RBS-GFP-TT Ant. enzymes Vector Insert Cloner 1.3a
Progress Notes
Controls W/ PQ (-) GFP tag w/ No Promoter RBS-GFP-TT Ant. enzymes Vector Insert Cloner 1.3a
Progress Notes
Controls Blh and Raldh II (+) Constitutive Promoter + Blh and RaldhII Promoter+ RBS-Blh-TT-RBS-RALDH II-TT Ant. enzymes Vector Insert Cloner 1.3a
Progress Notes
Controls Blh and Raldh II (-) Blh and RaldhII w/ no Promoter RBS-Blh-TT-RBS-RALDH II-TT Ant. enzymes Vector Insert Cloner 1.3a
Progress Notes

Device 1 Cloning Plan with Proper Sequences

Device 1 Cloning Plan with Proper Suffix

Cloning Step Basic Parts Detailed Parts Antibiotics Enzymes Vector & Size (kb) Insert & Size (kb) Current Cloner Picture
Step 1.1a SoxR/S Promoter + GFP tag RBS-SoxR-TT-SoxS Promoter + RBS-GFP-TT Ant. enzymes Vector ----Insert---- Cloner 1.1a
Progress Notes
Step 1.1b SoxR/S Promoter + Blh RBS-SoxR-TT-SoxS Promoter + RBS-Blh Ant. enzymes Vector Insert Cloner 1.1b
Progress Notes
Step 1.1c Blh + Terminator RBS-Blh + TT Ant. enzymes Vector Insert Cloner 1.1c
Progress Notes
Step 1.1d Constitutive Promoter + Blh (1.1c) Promoter +RBS-Blh-TT Ant. enzymes Vector Insert Cloner 1.1d
Progress Notes
Step 1.1e Constitutive Promoter + Crt Cluster Promoter + RBS-Crt Ant. enzymes Vector Insert Cloner 1.1e
Progress Notes
Step 1.2a Constitutive Promoter + SoxR/s w/ GFP (1.1a) Promoter+ RBS-SoxR-TT-SoxSP-RBS-GFP-TT Ant. enzymes Vector| Insert Cloner 1.2a
Progress Notes
Step 1.2b SoxR/S-Blh (1.1b) + RALDH II RBS-SoxR-TT-SoxSP-RBS-Blh + RBS-RALDH II-TT Ant. enzymes Vector Insert Cloner 1.2b
Progress Notes
Step 1.3a Constitutive Promoter + SoxR/S w/ Blh and RaldhII Promoter+ RBS-SoxR-TT-SoxSP-RBS-Blh-RBS-RALDH II-TT Ant. enzymes Vector Insert Cloner 1.3a
Progress Notes
Controls W/ PQ (+) Constitutive Promoter + GFP tag Promoter+ RBS-GFP-TT Ant. enzymes Vector Insert Cloner 1.3a
Progress Notes
Controls W/ PQ (-) GFP tag w/ No Promoter RBS-GFP-TT Ant. enzymes Vector Insert Cloner 1.3a
Progress Notes
Controls Blh and Raldh II (+) Constitutive Promoter + Blh and RaldhII Promoter+ RBS-Blh-TT-RBS-RALDH II-TT Ant. enzymes Vector Insert Cloner 1.3a
Progress Notes
Controls Blh and Raldh II (-) Blh and RaldhII w/ no Promoter RBS-Blh-TT-RBS-RALDH II-TT Ant. enzymes Vector Insert Cloner 1.3a
Progress Notes

Device 2 Cloning Plan with Transposed Suffix

Device 2 Cloning Plan with Transposed Suffix

Cloning Step Basic Parts Detailed Parts Antibiotics Enzymes Vector & Size (kb) Insert & Size (kb) Current Cloner Picture
Step 2.1a hIl-6-HlyAs + Terminator hIl-6-HlyAs + TT Ant. enzymes Vector ----Insert---- Cloner 1.1a
Progress Notes
Step 2.1b hIl-6-HlyAs + YFP hIl-6-HlyAs + YFP Ant. enzymes Vector Insert Cloner 1.1b
Progress Notes
Step 2.1c RBS + TolC RBS+ TolC Ant. enzymes Vector Insert Cloner 1.1c
Progress Notes
Step 2.1d Terminator + HlyR TT + HlyR Ant. enzymes Vector Insert Cloner 1.1d
Progress Notes
Step 2.1e Forward Flanking Sequence (FFS) + RBS Hly-C + RBS Ant. enzymes Vector Insert Cloner 1.1e
Progress Notes
Step 2.1f Rear Flanking Sequence (RFS) + Terminator HlyB,D + TT Ant. enzymes Vector Insert Cloner 1.1e
Progress Notes
Step 2.2a RBS + hIL-6-HlyAs (2.1a) RBS + hIL-6-HlyAs-TT Ant. enzymes Vector| Insert Cloner 1.2a
Progress Notes
Step 2.2b hIl-6-HlyAs-YFP(2.1b) + Terminator hIl-6-HlyAs-YFP+ TT Ant. enzymes Vector Insert Cloner 1.2b
Progress Notes
Step 2.2c TolC (2.1c) + HlyR Promoter Element (2.1d) RBS-TolC + TT-HlyR Ant. enzymes Vector Insert Cloner 1.2b
Progress Notes
Step 2.2d FFS-RBS (2.1e) + RFS-TT (2.1f) FFS-RBS + RFS-TT Ant. enzymes Vector Insert Cloner 1.2b
Progress Notes
Step 2.3a Trp Operator + hIL-6-HlyAs (2.2a) Trp Op + RBS-hIL-6-HlyAs-TT Ant. enzymes Vector Insert Cloner 1.3a
Progress Notes
Step 2.3b RBS + hIL-6-HlyAs-YFP-TT (2.2b) RBS + hIL-6-HlyAs-YFP-TT Ant. enzymes Vector Insert Cloner 1.3a
Progress Notes
Step 2.3c TolC-HlyR (2.2c) + FFS-RFS (2.2d) RBS-TolC-TT-HlyR + FFS-RBS-RFS-TT Ant. enzymes Vector Insert Cloner 1.3a
Progress Notes
Step 2.4a Trp Operator + HIl-6-HlyAs-YFP (2.3b) Trp Op + RBS-HIl-6-HlyAs-YFP-TT Ant. enzymes Vector Insert Cloner 1.3a
Progress Notes
Step 2.4b Const. Promoter + TolC-HlyR-FFS-RFS (2.3c) Const. Promoter + RBS-TolC-TT-HlyR-FFS-RBS-RFS-TT Ant. enzymes Vector Insert Cloner 1.3a
Progress Notes

Device 2 Cloning Plan with Proper Sequences

Device 2 Cloning Plan with Proper Suffix
Cloning Step Basic Parts Detailed Parts Antibiotics Enzymes Vector & Size (kb) Insert & Size (kb) Current Cloner Picture
Step 2.1a hIl-6-HlyAs + Terminator hIl-6-HlyAs + TT Ant. enzymes Vector ----Insert---- Cloner 1.1a
Progress Notes
Step 2.1b hIl-6-HlyAs + YFP hIl-6-HlyAs + YFP Ant. enzymes Vector Insert Cloner 1.1b
Progress Notes
Step 2.1c RBS + TolC RBS+ TolC Ant. enzymes Vector Insert Cloner 1.1c
Progress Notes
Step 2.1d Terminator + HlyR TT + HlyR Ant. enzymes Vector Insert Cloner 1.1d
Progress Notes
Step 2.1e Forward Flanking Sequence (FFS) + RBS Hly-C + RBS Ant. enzymes Vector Insert Cloner 1.1e
Progress Notes
Step 2.1f Rear Flanking Sequence (RFS) + Terminator HlyB,D + TT Ant. enzymes Vector Insert Cloner 1.1e
Progress Notes
Step 2.2a RBS + hIL-6-HlyAs (2.1a) RBS + hIL-6-HlyAs-TT Ant. enzymes Vector| Insert Cloner 1.2a
Progress Notes
Step 2.2b hIl-6-HlyAs-YFP(2.1b) + Terminator hIl-6-HlyAs-YFP+ TT Ant. enzymes Vector Insert Cloner 1.2b
Progress Notes
Step 2.2c TolC (2.1c) + HlyR Promoter Element (2.1d) RBS-TolC + TT-HlyR Ant. enzymes Vector Insert Cloner 1.2b
Progress Notes
Step 2.2d FFS-RBS (2.1e) + RFS-TT (2.1f) FFS-RBS + RFS-TT Ant. enzymes Vector Insert Cloner 1.2b
Progress Notes
Step 2.3a Trp Operator + hIL-6-HlyAs (2.2a) Trp Op + RBS-hIL-6-HlyAs-TT Ant. enzymes Vector Insert Cloner 1.3a
Progress Notes
Step 2.3b RBS + hIL-6-HlyAs-YFP-TT (2.2b) RBS + hIL-6-HlyAs-YFP-TT Ant. enzymes Vector Insert Cloner 1.3a
Progress Notes
Step 2.3c TolC-HlyR (2.2c) + FFS-RFS (2.2d) RBS-TolC-TT-HlyR + FFS-RBS-RFS-TT Ant. enzymes Vector Insert Cloner 1.3a
Progress Notes
Step 2.4a Trp Operator + HIl-6-HlyAs-YFP (2.3b) Trp Op + RBS-HIl-6-HlyAs-YFP-TT Ant. enzymes Vector Insert Cloner 1.3a
Progress Notes
Step 2.4b Const. Promoter + TolC-HlyR-FFS-RFS (2.3c) Const. Promoter + RBS-TolC-TT-HlyR-FFS-RBS-RFS-TT Ant. enzymes Vector Insert Cloner 1.3a
Progress Notes
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