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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. (Link opens in a new window) PubMed (Link opens in a new window) Article

Plasmids from Article

ID Plasmid Purpose
90491pPKm-118Expresses pcDNA3 - CMVmin - 5X UAS - pFR Luciferase
90492pPKm-202Expresses pcDNA3 - CMVmin - 5X UAS - pFR - CMVmin Luciferase
90493pPKm-102pcDNA3 - CMVmin - mOrange, plasmid expressing orange fluorescent protein (OFP)
90494pPKm-112pcDNA3 - pCMV - MTAD - PIF3, expresses Phytochrome interacting factor 3 (PIF3) and minimal transactivation domain (MTAD), under CMV promoter
90495pPKm-113pcDNA3 -pCMV - MTAD - PIF6, expresses the Phytochrome interacting factor 6 (PIF6) and MTAD, under CMV promoter
90496pPKm-195pcDNA3 - CMVmin - PhyB - MTAD, expresses PhyB and MTAD, under CMV promoter
90497pPKm- 226pcDNA3 - CMVmin - PIF3 - VPR expresses PIF3 and VPR transactivator domain , under CMV promoter
90498pPKm- 227pcDNA3 - CMVmin - VPR - PIF3, expresses VPR transactivator domain and PIF3, under CMV promoter
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.
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
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
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
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
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
90505pPKm-145Empty plasmid, pSIN-EF1-alpha-IRES-puro
90506pPKm-243mOrange and mitochondrial-tagged sfGFP (pSIN-OFP-mitosfGFP)
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
90508pPKm-233pSIN - EF-1alpha - sFD - P2A - MTS - sFNR, vector encoding for Synechococcus sp. Ferredoxin (Fd) and Ferredoxin-oxireductase (FNR)
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)
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
90511pPKm-196pcDNA3 - CMVmin PIF6-DBD, expressing PIF6-DBD under CMV promoter
104626pPKm-300Expresses pSIN - EF-1alpha - MTS - tFd, vector encoding mitochondrial-tagged Thermosynechococcus elongates Ferredoxin (Fd), under EF-1alpha promoter
104853pPKm-105pcDNA3 - pCMV - PhyB NT - GBD, expressing Phytochrome B (PhyB) with Gal Binding Domain (GBD), under CMV promoter
104854pPKm-163pcDNA3 - pCMV - PIF3 - GBD, expressing PIF3 and GBD, under CMV promoter
104855pPKm-241pSIN-cytosFD-P2A-cytosFNR, expressing Synechococcus sp Fd and FNR, under EF-1alpha promoter
105816pPKm-292pcDNA - GAL4 DBD - MTAD. Expresses Gal4 DNA binding domain fused to MTAD
105817pPKm-293pcDNA - TET DBD - MTAD. Expresses TET DNA binding domain fused to MTAD

Antibodies from Article