pMJ505: pLM005-Cre12.g498550-Venus-3xFLAG-3xCtermSAGA2

Description: pMJ504, containing the gDNA of Cre12.g498550, was reengineered by restriction digestion and T4 ligation to add in-frame immediately after the 3xFLAG 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 to produce CC-5650

Plasmid: 9951 bp DNA circular
CDS	0784..1587	“APHVIII"
CDS	2699..5694	“MPM1”
CDS	5695..6774	“VENUS-3xFLAG-3xSAGA2Cterm fusion protein”
TRANSLATION: NGDLGGSGGRMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELIKGGGDYKDHDGDYKDHDIDYKDDDDKGGGGSGGGGSPVRRTAIPANWRDALGGGGSGGGGSGGGGSPVRRTAIPANWRDALGGGGSGGGGSGGGGSPVRRTAIPANWRDAL-

Plasmid sequence	1..9951
TCGCGCGTTTCGGTGATGACGGTGAAAACCTCTGACACATGCAGCTCCCGGAGACGGTCACAGCTTGTCTGTAAGCGGATGCCGGGAGCAGACAAGATATCCTTGACATGATTGGTGCGTATGTTTGTATGAAGCTACAGGACTGATTTGGCGGGCTATGAGGGCGGGGGAAGCTCTGGAAGGGCCGCGATGGGGCGCGCGGCGTCCAGAAGGCGCCATACGGCCCGCTGGCGGCACCCATCCGGTATAAAAGCCCGCGACCCCGAACGGTGACCTCCACTTTCAGCGACAAACGAGCACTTATACATACGCGACTATTCTGCCGCTATACATAACCACTCAGCTAGCTTAAGATCCCATCAAGCTTGCATGCCGGGCGCGCCAGAAGGAGCGCAGCCAAACCAGGATGATGTTTGATGGGGTATTTGAGCACTTGCAACCCTTATCCGGAAGCCCCCTGGCCCACAAAGGCTAGGCGCCAATGCAAGCAGTTCGCATGCAGCCCCTGGAGCGGTGCCCTCCTGATAAACCGGCCAGGGGGCCTATGTTCTTTACTTTTTTACAAGAGAAGTCACTCAACATCTTAAAATGGCCAGGTGAGTCGACGAGCAAGCCCGGCGGATCAGGCAGCGTGCTTGCAGATTTGACTTGCAACGCCCGCATTGTGTCGACGAAGGCTTTTGGCTCCTCTGTCGCTGTCTCAAGCAGCATCTAACCCTGCGTCGCCGTTTCCATTTGCAGGATGGCCACTCCGCCCTCCCCGGTGCTGAAGAATTTCGAAGCATGGACGATGCGTTGCGTGCACTGCGGGGTCGGTATCCCGGTTGTGAGTGGGTTGTTGTGGAGGATGGGGCCTCGGGGGCTGGTGTTTATCGGCTTCGGGGTGGTGGGCGGGAGTTGTTTGTCAAGGTGGCAGCTCTGGGGGCCGGGGTGGGCTTGTTGGGTGAGGCTGAGCGGCTGGTGTGGTTGGCGGAGGTGGGGATTCCCGTACCTCGTGTTGTGGAGGGTGGTGGGGACGAGAGGGTCGCCTGGTTGGTCACCGAAGCGGTTCCGGGGCGTCCGGCCAGTGCGCGGTGGCCGCGGGAGCAGCGGCTGGACGTGGCGGTGGCGCTCGCGGGGCTCGCTCGTTCGCTGCACGCGCTGGACTGGGAGCGGTGTCCGTTCGATCGCAGTCTCGCGGTGACGGTGCCGCAGGCGGCCCGTGCTGTCGCTGAAGGGAGCGTCGACTTGGAGGATCTGGACGAGGAGCGGAAGGGGTGGTCGGGGGAGCGGCTTCTCGCCGAGCTGGAGCGGACTCGGCCTGCGGACGAGGATCTGGCGGTTTGCCACGGTGACCTGTGCCCGGACAACGTGCTGCTCGACCCTCGTACCTGCGAGGTGACCGGGCTGATCGACGTGGGGCGGGTCGGCCGTGCGGACCGGCACTCCGATCTCGCGCTGGTGCTGCGCGAGCTGGCCCACGAGGAGGACCCGTGGTTCGGGCCGGAGTGTTCCGCGGCGTTCCTGCGGGAGTACGGGCGCGGGTGGGATGGGGCGGTATCGGAGGAAAAGCTGGCGTTTTACCGGCTGTTGGACGAGTTCTTCTGAGGGACCTGATGGTGTTGGTGGCTGGGTAGGGTTGCGTCGCGTGGGTGACAGCACAGTGTGGACGTTGGGATCCCCGCTCCGTGTAAATGGAGGCGCTCGTTGATCTGAGCCTTGCCCCCTGACGAACGGCGGTGGATGGAAGATACTGCTCTCAAGTGCTGAAGCGGTAGCTTAGCTCCCCGTTTCGTGCTGATCAGTCTTTTTCAACACGTAAAAAGCGGAGGAGTTTTGCAATTTTGTTGGTTGTAACGATCCTCCGTTGATTTTGGCCTCTTTCTCCATGGGCGGGCTGGGCGTATTTGAAGCGGGTACCAGCTTTTGTTCCCTTTAGTGAGGGTTAATTGCGACCAATCGTCACACGAGCCCTCGTCAGAAACACGTCTCCGCCACGCTCTCCCTCTCACGGCCGACCCCGCAGCCCTTTTGCCCTTTCCTAGGCCACCGACAGGACCCAGGCGCTCTCAGCATGCCTCAACAACCCGTACTCGTGCCAGCGGTGCCCTTGTGCTGGTGATCGCTTGGAAGCGCATGCGAAGACGAAGGGGCGGAGCAGGCGGCCTGGCTGTTCGAAGGGCTCGCCGCCAGTTCGGGTGCCTTTCTCCACGCGCGCCTCCACACCTACCGATGCGTGAAGGCAGGCAAATGCTCATGTTTGCCCGAACTCGGTGTCCTTAAAAAGCCGCTTCTTGTCGTCGTTCCGAGACATGTTAGCAGATCGCAGTGCCACCTTTCCTGACGCGCTCGGCCCCATATTCGGACGCAATTGTCATTTGTAGCACAATTGGAGCAAATCTGGCGAGGCAGTAGGCTTTTAAGTTGCAAGGCGAGAGAGCAAAGTGGGACGCGGCGTGATTATTGGTATTTACGCGACGGCCCGGCGCGTTAGCGGCCCTTCCCCCAGGCCAGGGACGATTATGTATCAATATTGTTGCGTTCGGGCACTCGTGCGAGGGCTCCTGCGGGCTGGGGAGGGGGATCTGGGAATTGGAGGTACGACCGAGATGGCTTGCTCGGGGGGAGGTTTCCTCGCCGAGCAAGCCAGGGTTAGGTGTTGCGCTCTTGACACGTTGTGCATTCTAGGACCCCACTGCTACTCACAACAAGCCCAGTTATGCAGTGCCTAAACACCAAGCCTTTTACCTCGGCCCGCCCGGCTGTGGCCCGGCGGAGCGTCGTCGCGCATGCCTCGCGCCCGGCCCAGGCTGCCGCGCTGGCCTCTGCCATGACCGTGGCGACCACCTCCGCTGCTATGGCTTCTGAGATTGCTCAAACTGCCGATGTTGGCTCGCTGACCTTCGCGGTTGGCGGAGTGGGCGCTGTTGTGGGCCTGGGCGCCCTGCTTGTCGCTACTGACCCTCAGAAGCGGTGAGAAGGGTGCAGGAGTGCCTCAGATCTTAATGCAGATGCGCGTGCTTGGCTAGGATGGCATCACGCAAGCAAACATCGCGTTTCTGCTCCTCGCAGCCGCTCGGAGCAGATGAAGTCATTCGACGGCGATGAGAAGGAGGCGGTGAGTCCAAAATACAGGCTTCAATTTGAGTTGGAGGCAGCAATCCGACCCTGCGCCCCAGCTGTGTTCTGCGCCGGGTATGGGGACGTGCCACCCCTCTCCGCGGATCCGGTCTTCTCACGGACTGGGTTTGCTGTGAATTGTCCGGACAGCCACCGACGAAGTTCAGCATCATCTACCGCTACCCGGATACCCTGGCAGATCTAGTGTTGCAGCTTGATATCAGCAGCTAGCGCGACAACCACGCCAGCAACAGCCACGTCGCTCCGCGCACTCGCGCCCGCAGCGACCACCTCCCCTCACCCCCTCCCCCCATCACCCCATCAGGTGAAGGACTACTTCAACACTGCCGGCTTCGAGCGCTGGCGCAAGATCTACGGCGAGACGGACGAGGTGAACAAGGTGCAGCTGGACATCCGCACTGGCCACGCGCAGACGGTGGACAAGGTGCTGCGCTGGGTGGACGAGGAGGGCAGCGTGCAGGGCATCACCGTGGCGGACTGCGGCTGCGGGACAGGTGGGAGGGGCAGAGGGCCGTGGGCCGGGGGGTAGAAGGAGGGGCAGGAGGAGGGGAGGAGGCTGTCGCGGGGATGGCGCTGGTTGGCGTGCGGGTAGAGGGTAAGAGGGCGGAAGGGGCTGGGGCAGGTGAACGTGTTTGGAAAGGTTGCAGGAAGCACGGACATGGTGCACGGACAGAAGGCTTATGATGACGCAATAGCAGGGCGTGCCGAGTCGTGGGCGTACCCCGCGGCCAAGAATGCAATTCGGCAGCGCCAAGCAGTTTGCTTGGCAAGCCAGCACTATGGTATTCCCACCGCCGGAGGCGACGACCCACACATGCTTAGCACGACCTCTCCCGGCAACTCATGACCGCCTCTCCTCCCGCTCCCTCTGCCCCTCGGCACACCTCCTCCCAACACACCACCCACAGGCAGCCTGGCCATCCCGCTGGCGCTGCGCGGCGCCGCCGTCAGCGCCTCCGACATCTCCGCCGCCATGGCGTCTGAGGCGGAGCAGCGCTACCAGCAGGCGGTGGCGGCCGGGCAGGGCAAGGCGCCCAAGGTGGCGCCCAAGTTCGAGGCGCTGGACCTGGAGTGAGTGCTGGGGAGAGAGAGGACGGGAGAGGGAGGGCGCGAGGAAGGAACGAGGGAACAAAGGGGAGGGGGATAGGAGAACTGGTGGTTGGAACAAAAGCACGCGAACACAATGGAGGCGGGAAGCAGATCAGGCGGGGAAATATGAAAGCGGAGGAGGGTAGGAGGGCCGAGGTCAGGGGAGGAAGGAAAAGGGCCGCAAGACGGGAGGCGCTGTCGGGCATGGAAGCTGAATGGAGGACGGCAAGAAGTGTCTGCGGGCGCGCCGTGAATGGACACGGTGGTGGTGGTGGGGGGGGGGGGTAGTTCATTCCACTTCCATTATTAACCACTGCCAATCGGGGAAGAGTGCGAGGGCGGAAAGGGATGAGGGCCAGCCAATGGCTCCAGCGCTGTTGTTCACGAGGCGACCAGCGCGGCTGGAGGGGACCATCATCACATCATGGCGAGCGCAGGGCGTAGGACTCCGTGGAACGGAACGGGACGGGAAAAGGGACGGGGTCGGAAGTCGGGTTGGAGGGGGCCCGCGACCGAGCGCGGGCGAGGGCTGCCGTGCGTGCGTGTTTCCGAACCCCAACCGACCAAGCTGTCGCCTTTCCGCACCCGCCCTCACTGCCGCAGGTCCGTCAAGGGCAAGTACGACACGGTCACTTGCCTGGACGTGATGATTCACTACCCCCAGGTGCGGCCGGGGCCCGGTGCGGATTGGGCGGGCGTGTCGTTTGCACACGTTGTGTGTTGGAAAGAGTCAACTGCTAGCCCACATGAACCCATACAAGCCGTGGGGAAATGTGGCGAGTGCAATTGTATGTCACATATGCCATTTGGGGCGGGGAGGGCGGGGCTGCTGACAGTGGTGGACTGGGCTGCAGCGTTTGGCGGTGACTGAAATGGCACACCGGTGCAGGCCCCTGTGCCACAGCCGCGTGCGCTTTGAAGCTCGGGGACAGCCCTCACAGCAGCATCGTCTGGCCGCGCTCCGCCTGCTTACGCCGTTTCCTCTCTCCCGCGCCCTCCTCACCCCGCAGGACAAGGTGGACGCCATGATCACGCACCTGGCGGGTCTGTCCGACCGCCGCCTCATCATCTCGTTCGCACCCAAGACGCTGTCGTACAGCATCCTCAAGCGCATTGGCGAGCTGTTCCCCGGCCCCAGCAAGGTGCGCAGCCTGCGAAGGGGCAGGGCTGATGTGCACTTATTGGGGTGCAATGACGAGCTGACGTTGTTGTTCCAAGTGGTTGCTACCGGCGGTGCATGGCACTGGGATGTGGGGGCACACGGCGCCTTGAGGTGTCGCCGATGCTACCGCATGTCGCAGCGTTGTTACACCGTGTGGCCCCTCGTCCCACCCATGGCCCGCCTTCGTGCAGGCCACCCGCGCCTACCTGCACCGTGAGGAGGACGTGGAGGCTGCCCTCAAGCGCGCCGGCTTCAAGGTCACCAAGCGTGAGATGACCGCCACCAGCTTCTACTTCTCGCGCCTGCTGGAGGCCATCCGCGAGAACGGAGATCTGGGTGGCTCCGGCGGCCGGATGGTGAGCAAGGGCGAGGAGCTGTTCACCGGCGTGGTGCCCATCCTGGTGGAGCTGGACGGCGACGTGAACGGCCACAAGTTCAGCGTGAGCGGCGAGGGCGAGGGCGACGCCACCTACGGCAAGCTGACCCTGAAGCTGATCTGCACCACCGGCAAGCTGCCCGTGCCCTGGCCCACCCTGGTGACCACCCTGGGCTACGGCCTGCAGTGCTTCGCCCGCTACCCCGACCACATGAAGCAGCACGACTTCTTCAAGAGCGCCATGCCCGAGGGCTACGTGCAGGAGCGCACCATCTTCTTCAAGGACGACGGCAACTACAAGACCCGCGCCGAGGTGAAGTTCGAGGGCGACACCCTGGTGAACCGCATCGAGCTGAAGGGCATCGACTTCAAGGAGGACGGCAACATCCTGGGCCACAAGCTGGAGTACAACTACAACAGCCACAACGTGTACATCACCGCCGACAAGCAGAAGAACGGCATCAAGGCCAACTTCAAGATCCGCCACAACATCGAGGACGGCGGCGTGCAGCTGGCCGACCACTACCAGCAGAACACCCCCATCGGCGACGGCCCCGTGCTGCTGCCCGACAACCACTACCTGAGCTACCAGAGCGCCCTGAGCAAGGACCCCAACGAGAAGCGCGACCACATGGTGCTGCTGGAGTTCGTGACCGCCGCCGGCATCACCCTGGGCATGGACGAGCTGATCAAGGGCGGTGGCGACTACAAGGACCATGACGGTGACTATAAGGATCACGACATCGACTACAAGGACGATGACGACAAGGGCGGCGGCGGCTCGGGCGGCGGCGGCAGCCCCGTGCGCCGCACCGCTATCCCGGCCAACTGGCGTGACGCACTGGGCGGCGGCGGCTCGGGCGGCGGCGGCTCGGGCGGCGGCGGCAGCCCCGTGCGCCGCACCGCTATCCCGGCCAACTGGCGTGACGCACTGGGCGGCGGCGGCTCGGGCGGCGGCGGCTCGGGCGGCGGCGGCAGCCCCGTGCGCCGCACCGCTATCCCGGCCAACTGGCGTGACGCACTGTAAGATCTCTAACTGCACGAATTCTCAGGTCGGGACATCCCCGACCAGGGGCGGCGGGGATGTTGCTGGGCCGATGGAAAGTAGCAACCCAGCCAGCGGCTTCCAGCGCACTCCAGCTGCTCACGGTTGCGACATTGCGCGTGCACGCTTGCGCGTCCCTCACTCGGCCAGCTTGTCGCCGCAGACATCCCTAGCATTGTGCGGACTGCGGTCGTCAGTTAGCGTAGTGGCGGGGCTCAAAGCGTGATGCAGCTGGTGGCTGATTGCATGTGCTACATATGCTGTTATGTTTTGCATGAACTTCGATGCATTGGATGCTGGGTGCACGCGTTTGCATGTGTTTGTGCCGGCATGCTGCCGTCGTCGGCCGTACGTTTACGTTTCTGTGTGCCGGGGTCTTTATTTCCGCCTGCAGACCGACGTCGGTTCCACTAGTTCTAGATGGCAGCAGCTGGACCGCCTGTACCATGGAGAAGAGCTTTACTTGCCGGGATGGCCGATTTCGCTGATTGATACGGGATCGGAGCTCGGAGGCTTTCGCGCTAGGGGCTAGGCGAAGGGCAGTGGTGACCAGGGTCGGTGTGGGGTCGGCCCACGGTCAATTAGCCACAGGAGGATCAGGGGGAGGTAGGCACGTCGACTTGGTTTGCGACCCCGCAGTTTTGGCGGACGTGCTGTTGTAGATGTTAGCGTGTGCGTGAGCCAGTGGCCAACGTGCCACACCCATTGAGAAGACCAACCAACTTACTGGCAATATCTGCCAATGCCATACTGCATGTAATGGCCAGGCCATGTGAGAGTTTGCCGTGCCTGCGCGCGCCCCGGGGGCGAAGCTTGGTACCGCTAGCGTCAGCTGAACTCCTGCTGCCACGCTGACAGAGACGCGCCCATGACGTAGCGCCGACTGCTGCTCCTCTGCTCGTGGAGCTCTGAATCTGCTGCCGGCTGCCCCCGCCCCCGCTGCTGAGGCTGAATCTACTCCGCTCTCCACCCCAGTCCCAGGAAGAGAGGCGCATTTACTTCGCACAGACGTTACAGCACACCCTTGATCATCATCAGCTGCTCTTCCCTGCCGCTGCAACACGCCCCGCGCGATATCCCAGTCGGGAAACCTGTCGTGCCAGCTGCATTAATGAATCGGCCAACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGAACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTACAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAATACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAAAATAGGCGTATCACGAGGCCCTTTCGTC