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Biosynthesis of Orthogonal Molecules Using Ferredoxin and Ferredoxin-NADP(+) Reductase Systems Enables Genetically Encoded PhyB Optogenetics.
Kyriakakis P, Catanho M, Hoffner N, Thavarajah W, Hu VJ, Chao SS, Hsu A, Pham V, Naghavian L, Dozier LE, Patrick GN, Coleman TP
ACS Synth Biol. 2018 Feb 16;7(2):706-717. doi: 10.1021/acssynbio.7b00413. Epub 2018 Jan 24.
PubMed Article

Plasmids from Article

ID Plasmid Purpose  
90491pPKm-118Expresses pcDNA3 - CMVmin - 5X UAS - pFR Luciferase
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90492pPKm-202Expresses pcDNA3 - CMVmin - 5X UAS - pFR - CMVmin Luciferase
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90493pPKm-102pcDNA3 - CMVmin - mOrange, plasmid expressing orange fluorescent protein (OFP)
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90494pPKm-112pcDNA3 - pCMV - MTAD - PIF3, expresses Phytochrome interacting factor 3 (PIF3) and minimal transactivation domain (MTAD), under CMV promoter
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90495pPKm-113pcDNA3 -pCMV - MTAD - PIF6, expresses the Phytochrome interacting factor 6 (PIF6) and MTAD, under CMV promoter
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90496pPKm-195pcDNA3 - CMVmin - PhyB - MTAD, expresses PhyB and MTAD, under CMV promoter
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90497pPKm- 226pcDNA3 - CMVmin - PIF3 - VPR expresses PIF3 and VPR transactivator domain , under CMV promoter
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90498pPKm- 227pcDNA3 - CMVmin - VPR - PIF3, expresses VPR transactivator domain and PIF3, under CMV promoter
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90499pPKm-230pSIN - EF-1alpha - PIF3 - MTAD - IRES - PhyB - GBD, dual vector of PIF3-MTAD under EF-1 alpha promoter and PhyB-DBD under IRES promoter; See growth conditions below.
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90500pPKm-231pSIN - EF-1alpha - MTS - tFd - P2A - MTS - tFNR, vector encoding for mitochondrial-tagged Thermosynechococcus elongates Ferredoxin (Fd) and Ferredoxin-NADP(+) oxireductase (FNR), under EF-1alpha promoter
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90501pPKm-232pSIN - EF-1alpha - MTS tHO1 - P2A - MTS - tPCYA, vector encoding for mitochondrial-tagged Thermosynechococcus elongates Heme Oxigenase-1 (HO1) and Phycocyanobilin:ferredoxin oxireductase, under EF-1alpha promoter
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90502pPKm-244Expresses HO1, PCYA, FD and FNR, required for PCB synthesis. pSIN - EF-1alpha - MTS - tHO1 - P2A - MTS - tPCYA - IRES - MTS - tFD - P2A - MTS - tFNR
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90503pPKm-245Expresses HO1, PCYA, FD and FNR, required for PCB synthesis. pSIN - EF-1alpha - MTS - tHO1 - P2A - MTS - tPCYA - P2A - MTS - tFD - P2A - MTS - tFNR
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90504pPKm-248Expresses HO1, PCYA, FD and FNR, required for PCB synthesis. pSIN - EF-1alpha - MTS - tPCYA - IRES - MTS - tHO1 - P2A - MTS - tFD - P2A - MTS - tFNR
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90505pPKm-145Empty plasmid, pSIN-EF1-alpha-IRES-puro
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90506pPKm-243mOrange and mitochondrial-tagged sfGFP (pSIN-OFP-mitosfGFP)
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90507pPKm-234pSIN - EF-1alpha - MTS sHO1 - P2A - MTS - sPCYA, vector encoding for mitochondrial-tagged Synechococcus sp. Heme Oxigenase-1 (HO1) and Phycocyanobilin:ferredoxin oxireductase (PcyA), under EF-1alpha promoter
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90508pPKm-233pSIN - EF-1alpha - sFD - P2A - MTS - sFNR, vector encoding for Synechococcus sp. Ferredoxin (Fd) and Ferredoxin-oxireductase (FNR)
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90509pPKm-235pSIN - EF-1alpha - MTS sHO1 - P2A - MTS - Hy2, vector encoding for mitochondrial-tagged Synechococcus sp. Heme Oxigenase-1 (HO1) and phytochromobilin:ferredoxin oxidoreductase (Hy2)
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90510pPKm-240pSIN - EF-1alpha - Cyto -sFd - P2A - Cyto - sFNR, vector encoding for cytoplasmic-tagged Synechococcus sp. Heme Oxigenase-1 (HO1) and Phycocyanobilin:ferredoxin oxireductase (PcyA), under EF-1alpha promoter
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90511pPKm-196pcDNA3 - CMVmin PIF6-DBD, expressing PIF6-DBD under CMV promoter
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104626pPKm-300Expresses pSIN - EF-1alpha - MTS - tFd, vector encoding mitochondrial-tagged Thermosynechococcus elongates Ferredoxin (Fd), under EF-1alpha promoter
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104853pPKm-105pcDNA3 - pCMV - PhyB NT - GBD, expressing Phytochrome B (PhyB) with Gal Binding Domain (GBD), under CMV promoter
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104854pPKm-163pcDNA3 - pCMV - PIF3 - GBD, expressing PIF3 and GBD, under CMV promoter
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104855pPKm-241pSIN-cytosFD-P2A-cytosFNR, expressing Synechococcus sp Fd and FNR, under EF-1alpha promoter
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105816pPKm-292pcDNA - GAL4 DBD - MTAD. Expresses Gal4 DNA binding domain fused to MTAD
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105817pPKm-293pcDNA - TET DBD - MTAD. Expresses TET DNA binding domain fused to MTAD
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