pMJ503: pLM005-Cre14.g626700-Venus-3xFLAG-3xCtermSAGA2 Description: FDX1 (Cre14.g626700) gDNA coding sequence, spanning the start codon to the triplet just before the stop codon, had been previously cloned in frame into pLM005 by Gibson assembly (as described in https://doi.org/10.1073/pnas.1522866113; CSI_FC1F06 = pLM005-Cre14.g626700-Venus-3xFLAG). That plasmid was then reengineered by restriction digestion and T4 ligation to add in-frame immediately after the 3xFLAG and before the stop codon a synthetic fragment containing three copies of the sequence coding for the 15 C-terminal amino acids of SAGA2 (Cre 09.g394621), interspersed with a short flexible linker (GGGGGS)2. Selection in E. coli: ampicillin Selection in C. reinhardtii: paromomycin Host strain: DH5α Transformed into C. reinhardtii strain cMJ030 with carrier DNA to produce CC-5648 Plasmid: 7798 bp DNA circular CDS 784..1587 “APHVIII" CDS 2699..3541 “FDX1” CDS 3542..4621 “VENUS-3xFLAG-3xSAGA2Cterm fusion protein” TRANSLATION: NGDLGGSGGRMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELIKGGGDYKDHDGDYKDHDIDYKDDDDKGGGGSGGGGSPVRRTAIPANWRDALGGGGSGGGGSGGGGSPVRRTAIPANWRDALGGGGSGGGGSGGGGSPVRRTAIPANWRDAL- Plasmid sequence 1..7798 TCGCGCGTTTCGGTGATGACGGTGAAAACCTCTGACACATGCAGCTCCCGGAGACGGTCACAGCTTGTCTGTAAGCGGATGCCGGGAGCAGACAAGATATCCTTGACATGATTGGTGCGTATGTTTGTATGAAGCTACAGGACTGATTTGGCGGGCTATGAGGGCGGGGGAAGCTCTGGAAGGGCCGCGATGGGGCGCGCGGCGTCCAGAAGGCGCCATACGGCCCGCTGGCGGCACCCATCCGGTATAAAAGCCCGCGACCCCGAACGGTGACCTCCACTTTCAGCGACAAACGAGCACTTATACATACGCGACTATTCTGCCGCTATACATAACCACTCAGCTAGCTTAAGATCCCATCAAGCTTGCATGCCGGGCGCGCCAGAAGGAGCGCAGCCAAACCAGGATGATGTTTGATGGGGTATTTGAGCACTTGCAACCCTTATCCGGAAGCCCCCTGGCCCACAAAGGCTAGGCGCCAATGCAAGCAGTTCGCATGCAGCCCCTGGAGCGGTGCCCTCCTGATAAACCGGCCAGGGGGCCTATGTTCTTTACTTTTTTACAAGAGAAGTCACTCAACATCTTAAAATGGCCAGGTGAGTCGACGAGCAAGCCCGGCGGATCAGGCAGCGTGCTTGCAGATTTGACTTGCAACGCCCGCATTGTGTCGACGAAGGCTTTTGGCTCCTCTGTCGCTGTCTCAAGCAGCATCTAACCCTGCGTCGCCGTTTCCATTTGCAGGATGGCCACTCCGCCCTCCCCGGTGCTGAAGAATTTCGAAGCATGGACGATGCGTTGCGTGCACTGCGGGGTCGGTATCCCGGTTGTGAGTGGGTTGTTGTGGAGGATGGGGCCTCGGGGGCTGGTGTTTATCGGCTTCGGGGTGGTGGGCGGGAGTTGTTTGTCAAGGTGGCAGCTCTGGGGGCCGGGGTGGGCTTGTTGGGTGAGGCTGAGCGGCTGGTGTGGTTGGCGGAGGTGGGGATTCCCGTACCTCGTGTTGTGGAGGGTGGTGGGGACGAGAGGGTCGCCTGGTTGGTCACCGAAGCGGTTCCGGGGCGTCCGGCCAGTGCGCGGTGGCCGCGGGAGCAGCGGCTGGACGTGGCGGTGGCGCTCGCGGGGCTCGCTCGTTCGCTGCACGCGCTGGACTGGGAGCGGTGTCCGTTCGATCGCAGTCTCGCGGTGACGGTGCCGCAGGCGGCCCGTGCTGTCGCTGAAGGGAGCGTCGACTTGGAGGATCTGGACGAGGAGCGGAAGGGGTGGTCGGGGGAGCGGCTTCTCGCCGAGCTGGAGCGGACTCGGCCTGCGGACGAGGATCTGGCGGTTTGCCACGGTGACCTGTGCCCGGACAACGTGCTGCTCGACCCTCGTACCTGCGAGGTGACCGGGCTGATCGACGTGGGGCGGGTCGGCCGTGCGGACCGGCACTCCGATCTCGCGCTGGTGCTGCGCGAGCTGGCCCACGAGGAGGACCCGTGGTTCGGGCCGGAGTGTTCCGCGGCGTTCCTGCGGGAGTACGGGCGCGGGTGGGATGGGGCGGTATCGGAGGAAAAGCTGGCGTTTTACCGGCTGTTGGACGAGTTCTTCTGAGGGACCTGATGGTGTTGGTGGCTGGGTAGGGTTGCGTCGCGTGGGTGACAGCACAGTGTGGACGTTGGGATCCCCGCTCCGTGTAAATGGAGGCGCTCGTTGATCTGAGCCTTGCCCCCTGACGAACGGCGGTGGATGGAAGATACTGCTCTCAAGTGCTGAAGCGGTAGCTTAGCTCCCCGTTTCGTGCTGATCAGTCTTTTTCAACACGTAAAAAGCGGAGGAGTTTTGCAATTTTGTTGGTTGTAACGATCCTCCGTTGATTTTGGCCTCTTTCTCCATGGGCGGGCTGGGCGTATTTGAAGCGGGTACCAGCTTTTGTTCCCTTTAGTGAGGGTTAATTGCGACCAATCGTCACACGAGCCCTCGTCAGAAACACGTCTCCGCCACGCTCTCCCTCTCACGGCCGACCCCGCAGCCCTTTTGCCCTTTCCTAGGCCACCGACAGGACCCAGGCGCTCTCAGCATGCCTCAACAACCCGTACTCGTGCCAGCGGTGCCCTTGTGCTGGTGATCGCTTGGAAGCGCATGCGAAGACGAAGGGGCGGAGCAGGCGGCCTGGCTGTTCGAAGGGCTCGCCGCCAGTTCGGGTGCCTTTCTCCACGCGCGCCTCCACACCTACCGATGCGTGAAGGCAGGCAAATGCTCATGTTTGCCCGAACTCGGTGTCCTTAAAAAGCCGCTTCTTGTCGTCGTTCCGAGACATGTTAGCAGATCGCAGTGCCACCTTTCCTGACGCGCTCGGCCCCATATTCGGACGCAATTGTCATTTGTAGCACAATTGGAGCAAATCTGGCGAGGCAGTAGGCTTTTAAGTTGCAAGGCGAGAGAGCAAAGTGGGACGCGGCGTGATTATTGGTATTTACGCGACGGCCCGGCGCGTTAGCGGCCCTTCCCCCAGGCCAGGGACGATTATGTATCAATATTGTTGCGTTCGGGCACTCGTGCGAGGGCTCCTGCGGGCTGGGGAGGGGGATCTGGGAATTGGAGGTACGACCGAGATGGCTTGCTCGGGGGGAGGTTTCCTCGCCGAGCAAGCCAGGGTTAGGTGTTGCGCTCTTGACACGTTGTGCATTCTAGGACCCCACTGCTACTCACAACAAGCCCAGTTATGACGGCCTCTTCAGCGCCTACTGTTGCGCCTACTGTAAAGCTGACCATCAACACGAAGGAGGGTCCGACAGTGGAATTGGACGTGGAGAGCGGGGACAAGCTTCGCGACGTGCTGATGGCCAACAAGGTGGAACTCTACACGACCTGGTGAGTTTAACCAATGGTCCCGCTCGGCCGCAGCGCCGGGTTTGCCTTGACACGGATGGTGCCGACGATGGCTGCCTTGCCACGCAGGGGAAAGGTGTGGTCTTGCGGCGGCGGCGGACAATGCGGCACGTGCATTGTTGACGTGAGTAAATACTGCGTGAAGACAAATATGGCTGACCTGCTTTGAGAGGGGTGGTGGCCTGGTGGGCTTCTCGCACTTAAAGGACAAGGATTGGGCAACGGCCACCGGCACGGGCCACCGGTACAGGTACCCCTGACGTGTTTGCACGTGCGGGGGCTGCCTGCAGGTGAAGAGTGGCGCTGACATCCTCACAGAACGCACCGGCGCAGAGAAGAAGAAGCTCTCTGGGGTGAGACATCAGTGGGGGAGGGCCTCTCTAGCATGCCAGGCGCCTGTGCACGCGTTGTGGGGCGGGGGCGGAAACACAGCGGGCTGAAACGCACACGATGTCAGATTGTTGAAATAGTGATGTGTGTGCATGCGACTTGAAGTTCCGGCCCGGCCCCGTGCATGCATCGTCACTACGAAAGCTTTCCTATCTCATCACGTACCCATCTTGTCCCGGGCCCGCCGCCTTGCTGACCGCAGAAGCCCGAGAGCTTCCGGTTGGCTTGCCAGACACTGGTGGGCGATGGCGAGAACAGCGGCGCCGTCACGATCGCAACCAAGCCCAACGGAGATCTGGGTGGCTCCGGCGGCCGGATGGTGAGCAAGGGCGAGGAGCTGTTCACCGGCGTGGTGCCCATCCTGGTGGAGCTGGACGGCGACGTGAACGGCCACAAGTTCAGCGTGAGCGGCGAGGGCGAGGGCGACGCCACCTACGGCAAGCTGACCCTGAAGCTGATCTGCACCACCGGCAAGCTGCCCGTGCCCTGGCCCACCCTGGTGACCACCCTGGGCTACGGCCTGCAGTGCTTCGCCCGCTACCCCGACCACATGAAGCAGCACGACTTCTTCAAGAGCGCCATGCCCGAGGGCTACGTGCAGGAGCGCACCATCTTCTTCAAGGACGACGGCAACTACAAGACCCGCGCCGAGGTGAAGTTCGAGGGCGACACCCTGGTGAACCGCATCGAGCTGAAGGGCATCGACTTCAAGGAGGACGGCAACATCCTGGGCCACAAGCTGGAGTACAACTACAACAGCCACAACGTGTACATCACCGCCGACAAGCAGAAGAACGGCATCAAGGCCAACTTCAAGATCCGCCACAACATCGAGGACGGCGGCGTGCAGCTGGCCGACCACTACCAGCAGAACACCCCCATCGGCGACGGCCCCGTGCTGCTGCCCGACAACCACTACCTGAGCTACCAGAGCGCCCTGAGCAAGGACCCCAACGAGAAGCGCGACCACATGGTGCTGCTGGAGTTCGTGACCGCCGCCGGCATCACCCTGGGCATGGACGAGCTGATCAAGGGCGGTGGCGACTACAAGGACCATGACGGTGACTATAAGGATCACGACATCGACTACAAGGACGATGACGACAAGGGCGGCGGCGGCTCGGGCGGCGGCGGCAGCCCCGTGCGCCGCACCGCTATCCCGGCCAACTGGCGTGACGCACTGGGCGGCGGCGGCTCGGGCGGCGGCGGCTCGGGCGGCGGCGGCAGCCCCGTGCGCCGCACCGCTATCCCGGCCAACTGGCGTGACGCACTGGGCGGCGGCGGCTCGGGCGGCGGCGGCTCGGGCGGCGGCGGCAGCCCCGTGCGCCGCACCGCTATCCCGGCCAACTGGCGTGACGCACTGTAAGATCTCTAACTGCACGAATTCTCAGGTCGGGACATCCCCGACCAGGGGCGGCGGGGATGTTGCTGGGCCGATGGAAAGTAGCAACCCAGCCAGCGGCTTCCAGCGCACTCCAGCTGCTCACGGTTGCGACATTGCGCGTGCACGCTTGCGCGTCCCTCACTCGGCCAGCTTGTCGCCGCAGACATCCCTAGCATTGTGCGGACTGCGGTCGTCAGTTAGCGTAGTGGCGGGGCTCAAAGCGTGATGCAGCTGGTGGCTGATTGCATGTGCTACATATGCTGTTATGTTTTGCATGAACTTCGATGCATTGGATGCTGGGTGCACGCGTTTGCATGTGTTTGTGCCGGCATGCTGCCGTCGTCGGCCGTACGTTTACGTTTCTGTGTGCCGGGGTCTTTATTTCCGCCTGCAGACCGACGTCGGTTCCACTAGTTCTAGATGGCAGCAGCTGGACCGCCTGTACCATGGAGAAGAGCTTTACTTGCCGGGATGGCCGATTTCGCTGATTGATACGGGATCGGAGCTCGGAGGCTTTCGCGCTAGGGGCTAGGCGAAGGGCAGTGGTGACCAGGGTCGGTGTGGGGTCGGCCCACGGTCAATTAGCCACAGGAGGATCAGGGGGAGGTAGGCACGTCGACTTGGTTTGCGACCCCGCAGTTTTGGCGGACGTGCTGTTGTAGATGTTAGCGTGTGCGTGAGCCAGTGGCCAACGTGCCACACCCATTGAGAAGACCAACCAACTTACTGGCAATATCTGCCAATGCCATACTGCATGTAATGGCCAGGCCATGTGAGAGTTTGCCGTGCCTGCGCGCGCCCCGGGGGCGAAGCTTGGTACCGCTAGCGTCAGCTGAACTCCTGCTGCCACGCTGACAGAGACGCGCCCATGACGTAGCGCCGACTGCTGCTCCTCTGCTCGTGGAGCTCTGAATCTGCTGCCGGCTGCCCCCGCCCCCGCTGCTGAGGCTGAATCTACTCCGCTCTCCACCCCAGTCCCAGGAAGAGAGGCGCATTTACTTCGCACAGACGTTACAGCACACCCTTGATCATCATCAGCTGCTCTTCCCTGCCGCTGCAACACGCCCCGCGCGATATCCCAGTCGGGAAACCTGTCGTGCCAGCTGCATTAATGAATCGGCCAACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGAACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTACAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAATACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAAAATAGGCGTATCACGAGGCCCTTTCGTC