pMJ508: pLM005-Cre10.g430350[W51A/R52A]-Venus-3xFLAG Description: Previously synthesized pMJ507 (CC-5652) was modified by site-directed mutagenesis. Triplet coding for Tryptophan in position 51 of the mature protein now codes for an Alanine. Triplet coding for Arginine in position 52 of the mature protein now codes for an Alanine. Selection in E. coli: ampicillin Selection in C. reinhardtii: paromomycin Host strain: DH5α Transformed into C. reinhardtii strain cMJ030 to produce CC-5653 Plasmid: 8646 bp DNA circular CDS 0784..1587 “APHVIII” CDS 2699..4644 “Cre10.g430350W51A/R52A” CDS 4645..5469 “VENUS-3xFLAG fusion protein” TRANSLATION: NGDLGGSGGRMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELIKGGGDYKDHDGDYKDHDIDYKDDDDK- Plasmid sequence: 1..8646 TCGCGCGTTTCGGTGATGACGGTGAAAACCTCTGACACATGCAGCTCCCGGAGACGGTCACAGCTTGTCTGTAAGCGGATGCCGGGAGCAGACAAGATATCCTTGACATGATTGGTGCGTATGTTTGTATGAAGCTACAGGACTGATTTGGCGGGCTATGAGGGCGGGGGAAGCTCTGGAAGGGCCGCGATGGGGCGCGCGGCGTCCAGAAGGCGCCATACGGCCCGCTGGCGGCACCCATCCGGTATAAAAGCCCGCGACCCCGAACGGTGACCTCCACTTTCAGCGACAAACGAGCACTTATACATACGCGACTATTCTGCCGCTATACATAACCACTCAGCTAGCTTAAGATCCCATCAAGCTTGCATGCCGGGCGCGCCAGAAGGAGCGCAGCCAAACCAGGATGATGTTTGATGGGGTATTTGAGCACTTGCAACCCTTATCCGGAAGCCCCCTGGCCCACAAAGGCTAGGCGCCAATGCAAGCAGTTCGCATGCAGCCCCTGGAGCGGTGCCCTCCTGATAAACCGGCCAGGGGGCCTATGTTCTTTACTTTTTTACAAGAGAAGTCACTCAACATCTTAAAATGGCCAGGTGAGTCGACGAGCAAGCCCGGCGGATCAGGCAGCGTGCTTGCAGATTTGACTTGCAACGCCCGCATTGTGTCGACGAAGGCTTTTGGCTCCTCTGTCGCTGTCTCAAGCAGCATCTAACCCTGCGTCGCCGTTTCCATTTGCAGGATGGCCACTCCGCCCTCCCCGGTGCTGAAGAATTTCGAAGCATGGACGATGCGTTGCGTGCACTGCGGGGTCGGTATCCCGGTTGTGAGTGGGTTGTTGTGGAGGATGGGGCCTCGGGGGCTGGTGTTTATCGGCTTCGGGGTGGTGGGCGGGAGTTGTTTGTCAAGGTGGCAGCTCTGGGGGCCGGGGTGGGCTTGTTGGGTGAGGCTGAGCGGCTGGTGTGGTTGGCGGAGGTGGGGATTCCCGTACCTCGTGTTGTGGAGGGTGGTGGGGACGAGAGGGTCGCCTGGTTGGTCACCGAAGCGGTTCCGGGGCGTCCGGCCAGTGCGCGGTGGCCGCGGGAGCAGCGGCTGGACGTGGCGGTGGCGCTCGCGGGGCTCGCTCGTTCGCTGCACGCGCTGGACTGGGAGCGGTGTCCGTTCGATCGCAGTCTCGCGGTGACGGTGCCGCAGGCGGCCCGTGCTGTCGCTGAAGGGAGCGTCGACTTGGAGGATCTGGACGAGGAGCGGAAGGGGTGGTCGGGGGAGCGGCTTCTCGCCGAGCTGGAGCGGACTCGGCCTGCGGACGAGGATCTGGCGGTTTGCCACGGTGACCTGTGCCCGGACAACGTGCTGCTCGACCCTCGTACCTGCGAGGTGACCGGGCTGATCGACGTGGGGCGGGTCGGCCGTGCGGACCGGCACTCCGATCTCGCGCTGGTGCTGCGCGAGCTGGCCCACGAGGAGGACCCGTGGTTCGGGCCGGAGTGTTCCGCGGCGTTCCTGCGGGAGTACGGGCGCGGGTGGGATGGGGCGGTATCGGAGGAAAAGCTGGCGTTTTACCGGCTGTTGGACGAGTTCTTCTGAGGGACCTGATGGTGTTGGTGGCTGGGTAGGGTTGCGTCGCGTGGGTGACAGCACAGTGTGGACGTTGGGATCCCCGCTCCGTGTAAATGGAGGCGCTCGTTGATCTGAGCCTTGCCCCCTGACGAACGGCGGTGGATGGAAGATACTGCTCTCAAGTGCTGAAGCGGTAGCTTAGCTCCCCGTTTCGTGCTGATCAGTCTTTTTCAACACGTAAAAAGCGGAGGAGTTTTGCAATTTTGTTGGTTGTAACGATCCTCCGTTGATTTTGGCCTCTTTCTCCATGGGCGGGCTGGGCGTATTTGAAGCGGGTACCAGCTTTTGTTCCCTTTAGTGAGGGTTAATTGCGACCAATCGTCACACGAGCCCTCGTCAGAAACACGTCTCCGCCACGCTCTCCCTCTCACGGCCGACCCCGCAGCCCTTTTGCCCTTTCCTAGGCCACCGACAGGACCCAGGCGCTCTCAGCATGCCTCAACAACCCGTACTCGTGCCAGCGGTGCCCTTGTGCTGGTGATCGCTTGGAAGCGCATGCGAAGACGAAGGGGCGGAGCAGGCGGCCTGGCTGTTCGAAGGGCTCGCCGCCAGTTCGGGTGCCTTTCTCCACGCGCGCCTCCACACCTACCGATGCGTGAAGGCAGGCAAATGCTCATGTTTGCCCGAACTCGGTGTCCTTAAAAAGCCGCTTCTTGTCGTCGTTCCGAGACATGTTAGCAGATCGCAGTGCCACCTTTCCTGACGCGCTCGGCCCCATATTCGGACGCAATTGTCATTTGTAGCACAATTGGAGCAAATCTGGCGAGGCAGTAGGCTTTTAAGTTGCAAGGCGAGAGAGCAAAGTGGGACGCGGCGTGATTATTGGTATTTACGCGACGGCCCGGCGCGTTAGCGGCCCTTCCCCCAGGCCAGGGACGATTATGTATCAATATTGTTGCGTTCGGGCACTCGTGCGAGGGCTCCTGCGGGCTGGGGAGGGGGATCTGGGAATTGGAGGTACGACCGAGATGGCTTGCTCGGGGGGAGGTTTCCTCGCCGAGCAAGCCAGGGTTAGGTGTTGCGCTCTTGACACGTTGTGCATTCTAGGACCCCACTGCTACTCACAACAAGCCCAGTTATGCCTATGCTTGGCTCTCGTCTTTTGGGTGCGCCCAGCACAAGCGCGCCAGCAAGGGTAAGTGGATCGACAGGTTGAATGCGATAGGCGGCATTCATACTGGCTCCGAAAGTTCGGTAGTGCTTCGGCGAGGTTCGGGACCCGAATGTTATCTGGAACCTGTGGTAACTGCTCACCGCGCGGCGTGATGAATCATCACAGGCTCCTCTGGCGCAGCATGGGCGACTTCTGGTGGCCTGCCGCGCGACTGTGAGGTCCAGGCAAGTGAAGCCGATTATTTCTTGTTGACTTCGCATATTCACGCGCGTTTCGGCATCAAAACGCGCTGGTGTGGGACCTTGGCTTCTCAGCTATTGCATAGCGCGCGGTGCGGGCGCCATCCAGGGCCTCTTCCTGTGTCCCGACCGCGCGGCTTCCAGCCACCGTGTGCACCGCAAACCTTGCTCCACGGTCTCCCTTAACAATTCCCGACTCGGCCCCGGCCCCAAACACAGCTCGCCACGCTCTACTGGCGACTGGCGCAGCTCCGCGGCAACGGAGCTTGAGGGCGGCGCCAACGGCGCGGCGCACCACGACCACGACCACGAGGAAGTCGACGTCCACGCCAACGGCCACAGCTACAGCAACGGCGTCTCCCATTCTGACAGCTACAGCAACGGCGCCAGCTACAACGGCACCAGTTACGCCACGGGCATCAGCGCCAGCTGGCGCAACGGCTCCGCCGCCGCCAACGGTAGCAGCTATGCGGCTCGCGCCGCGCAGCTGGTTCGCGAGCAGGACGAGGACGAGGACCACGACTTCCACCGGCGTGCAGCGGACTGGCGCACGGAGCACTCGCCGCAGGGCTCCCGCTGGGGCTCCGAGGACGACGATGCCGCCTCCTCCTCAGCCAACGGGCACCACCACGACCAGCAAGAGGAGGAGCAACAAGGTGAGCAGGAGGAAGGCGGCGGCGGTTTCTTCACGAGTGCCTTCAAGGACGCGCTGAAAGACGCCCTAGCGCGCAGTCCACTGTTCGGGTCGCAGTACGGCGGCGGGTCGGGCGACGAGGAGAGCATTCCGCCGCAGCCCACGGAGAACGAGGCGGTGGAGGCAACCACGGGTATGTGGTATTGTAGGGTGGTCTCTGCAGGGTCAAGGCAGGGGGTTGCGGCTTGGTGGGGCCAAAGGGCACGCAGGGCCTTCTCAACGACCATGAGCACGCCGTTGTAGGCTTTCCGCAATCCTGGTAAACTCCTGACAAGGTCATGACTGCCATACATCGCAACCTAAACACTAAACTCATCTTTCCTTCTGCGTGCAAACGTACTGTGGCAGGCTCGTCGCTGTTCAGCAACAGCTTCCGGTCTGCGCTAAAGGAGTCTCTGGAGCGGCAGCAGACGGAGATGGACGTCAAGTACGGGTACGGCAGCGATGACGAGTACATCGCCACGCCGTACAAGTCAGCGGGCCCGGCGCGCCGGTGAGTTAGAAGAAGCGGGCAGGGAGGGGGAGGAGGAGCAGGGGCAGGGAGGAGGACTGGTGGCGGGGGAGCGGGGCCTGGTCGTTCCGCACCCGAGGGCTCCAGGTGAGAGGTTGGCAGTACGACAGGGGACTGCCGCTGGCCTTGGGGTGTTTCCGAACACCGACCGGTCCTCTCAGCCGTCGCAAACGCTGGATACTGCACCACCGCGCTCACCCACCCGGCCCTCTGTCTGTTGCCCCTGCCTCTCCCTCTCCCACGTAGGGAGTTGTCCCCCATGGAGGAGCTGCTCTCTTCCGCGCTGAGCGCCAGCCAGCACACACACCGCCGCCTGACGCTCATAAAGAACGAGATCGATGCGGCGCTGGAGCGCGAGGCCAAGCAGATTGAGCGGCTGGATTTCGCTCTCAAGAAGTGCGAGAACGACGCGGCCTACTATGCGACGCTCGAGCGCATGATGCGCGAGCGCGAGCGGGAGGGCAAGAACGGAGATCTGGGTGGCTCCGGCGGCCGGATGGTGAGCAAGGGCGAGGAGCTGTTCACCGGCGTGGTGCCCATCCTGGTGGAGCTGGACGGCGACGTGAACGGCCACAAGTTCAGCGTGAGCGGCGAGGGCGAGGGCGACGCCACCTACGGCAAGCTGACCCTGAAGCTGATCTGCACCACCGGCAAGCTGCCCGTGCCCTGGCCCACCCTGGTGACCACCCTGGGCTACGGCCTGCAGTGCTTCGCCCGCTACCCCGACCACATGAAGCAGCACGACTTCTTCAAGAGCGCCATGCCCGAGGGCTACGTGCAGGAGCGCACCATCTTCTTCAAGGACGACGGCAACTACAAGACCCGCGCCGAGGTGAAGTTCGAGGGCGACACCCTGGTGAACCGCATCGAGCTGAAGGGCATCGACTTCAAGGAGGACGGCAACATCCTGGGCCACAAGCTGGAGTACAACTACAACAGCCACAACGTGTACATCACCGCCGACAAGCAGAAGAACGGCATCAAGGCCAACTTCAAGATCCGCCACAACATCGAGGACGGCGGCGTGCAGCTGGCCGACCACTACCAGCAGAACACCCCCATCGGCGACGGCCCCGTGCTGCTGCCCGACAACCACTACCTGAGCTACCAGAGCGCCCTGAGCAAGGACCCCAACGAGAAGCGCGACCACATGGTGCTGCTGGAGTTCGTGACCGCCGCCGGCATCACCCTGGGCATGGACGAGCTGATCAAGGGCGGTGGCGACTACAAGGACCATGACGGTGACTATAAGGATCACGACATCGACTACAAGGACGATGACGACAAGTAAGATCTCTAACTGCACGAATTCTCAGGTCGGGACATCCCCGACCAGGGGCGGCGGGGATGTTGCTGGGCCGATGGAAAGTAGCAACCCAGCCAGCGGCTTCCAGCGCACTCCAGCTGCTCACGGTTGCGACATTGCGCGTGCACGCTTGCGCGTCCCTCACTCGGCCAGCTTGTCGCCGCAGACATCCCTAGCATTGTGCGGACTGCGGTCGTCAGTTAGCGTAGTGGCGGGGCTCAAAGCGTGATGCAGCTGGTGGCTGATTGCATGTGCTACATATGCTGTTATGTTTTGCATGAACTTCGATGCATTGGATGCTGGGTGCACGCGTTTGCATGTGTTTGTGCCGGCATGCTGCCGTCGTCGGCCGTACGTTTACGTTTCTGTGTGCCGGGGTCTTTATTTCCGCCTGCAGACCGACGTCGGTTCCACTAGTTCTAGATGGCAGCAGCTGGACCGCCTGTACCATGGAGAAGAGCTTTACTTGCCGGGATGGCCGATTTCGCTGATTGATACGGGATCGGAGCTCGGAGGCTTTCGCGCTAGGGGCTAGGCGAAGGGCAGTGGTGACCAGGGTCGGTGTGGGGTCGGCCCACGGTCAATTAGCCACAGGAGGATCAGGGGGAGGTAGGCACGTCGACTTGGTTTGCGACCCCGCAGTTTTGGCGGACGTGCTGTTGTAGATGTTAGCGTGTGCGTGAGCCAGTGGCCAACGTGCCACACCCATTGAGAAGACCAACCAACTTACTGGCAATATCTGCCAATGCCATACTGCATGTAATGGCCAGGCCATGTGAGAGTTTGCCGTGCCTGCGCGCGCCCCGGGGGCGAAGCTTGGTACCGCTAGCGTCAGCTGAACTCCTGCTGCCACGCTGACAGAGACGCGCCCATGACGTAGCGCCGACTGCTGCTCCTCTGCTCGTGGAGCTCTGAATCTGCTGCCGGCTGCCCCCGCCCCCGCTGCTGAGGCTGAATCTACTCCGCTCTCCACCCCAGTCCCAGGAAGAGAGGCGCATTTACTTCGCACAGACGTTACAGCACACCCTTGATCATCATCAGCTGCTCTTCCCTGCCGCTGCAACACGCCCCGCGCGATATCCCAGTCGGGAAACCTGTCGTGCCAGCTGCATTAATGAATCGGCCAACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGAACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTACAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAATACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAAAATAGGCGTATCACGAGGCCCTTTCGTC