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A weak, faded, or off-hue solution can be an early qualitative flag that the copper occupancy or complex integrity is not what it should be, prompting a proper spectrophotometric check rather than a conclusion on its own
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Codon optimized GshF GCCGGATCTAGCTAGTTAATTAAGccaccatgACCCTTAATCAGCTTCTCCAGAAGTTGGAGGCGACT TCCCCCATTCTCCAGGCGAACTTCGGGATAGAAAGGGAGTCATTGAGGGTTGACCGCCAGGGTCA GCTGGTCCACACACCGCACCCCTCATGTCTGGGAGCCCGCAGTTTTCATCCTTACATACAAACCG ACTTTTGTGAATTCCAAATGGAACTGATTACACCAGTAGCCAAAAGTACGACGGAGGCCCGACGCT TTCTTGGCGCGATAACTGATGTAGCAGGACGAAGCATTGCAACTGACGAGGTGCTGTGGCCATTG AGTATGCCACCACGACTTAAAGCCGAGGAAATTCAAGTAGCGCAACTCGAGAACGACTTCGAAAG ACATTATCGGAACTACTTGGCAGAGAAGTACGGCACCAAATTGCAGGCGATTAGTGGAATTCATTA CAATATGGAACTTGGGAAGGACTTGGTTGAGGCGCTTTTTCAAGAGTCAGATCAGACTGACATGAT CGCATTTAAAAACGCTCTGTATCTCAAGCTCGCCCAGAACTATTTGAGGTATCGGTGGGTCATTAC TTACCTGTTTGGAGCAAGTCCCATTGCAGAACAAGGATTCTTTGACCAAGAAGTGCCGGAGCCTAT GCGCTCTTTCCGCAACTCCGACCACGGCTACGTTAACAAGGAAGAGATACAGGTAAGCTTTGTAT CCCTTGAAGACTATGTCTCCGCGATCGAGACCTACATCGAGCAGGGTGACCTTATAGCCGAGAAA GAGTTTTACTCAGCCGTGCGCTTTAGAGGACAAAAAGTCAATCGCTCCTTCCTTGATAAGGGTATA ACTTATCTGGAGTTCAGAAACTTTGACTTGAACCCATTTGAGAGAATAGGCATCAGTCAAACCACTA TGGATACCGTTCACTTGCTCATACTGGCCTTTCTCTGGTTGGATAGTCCGGAAAACGTGGACCAG GCCCTTGCCCAGGGACACGCGCTTAACGAGAAAATAGCCCTCTCCCATCCATTGGAGCCCCTCCC TTCAGAGGCCAAGACACAGGACATCGTGACCGCACTCGACCAGCTCGTACAGCACTTCGGATTGG GAGATTACCACCAGGATCTCGTGAAACAAGTGAAAGCGGCGTTTGCTGATCCGAATCAAACCCTG TCAGCTCAACTTCTTCCTTATATTAAGGACAAGTCACTCGCAGAATTCGCTCTCAATAAGGCACTCG CATATCATGACTATGACTGGACCGCTCACTACGCCCTTAAAGGTTACGAAGAAATGGAGCTCAGTA CGCAGATGCTGCTCTTTGATGCTATCCAGAAAGGAATACACTTCGAGATACTCGATGAGCAAGATC AGTTCTTGAAGCTGTGGCATCAAGATCACGTAGAATATGTTAAAAACGGTAATATGACCAGCAAGG ATAACTATGTAGTACCTCTCGCAATGGCCAACAAGACTGTTACTAAGAAAATTCTTGCTGACGCTG GGTTCCCTGTTCCGTCCGGGGACGAATTTACTAGCTTGGAGGAGGGACTGGCCTACTACCCGCTT ATTAAAGATAAGCAAATTGTAGTAAAGCCAAAGAGCACGAATTTCGGCTTGGGTATCAGCATCTTC CAAGAACCCGCCAGTCTCGACAATTATCAAAAAGCATTGGAAATAGCATTTGCGGAGGACACTAGT GTGCTCGTCGAAGAATTCATTCCAGGCACGGAATACCGATTCTTCATTTTGGACGGACGCTGTGA GGCAGTCCTTTTGAGGGTAGCTGCCAATGTAATAGGGGACGGGAAACACACAATCAGAGAGTTGG TAGCGCAGAAAAACGCAAATCCCCTGCGCGGTAGGGATCATAGATCACCCTTGGAAATCATAGAG CTTGGGGACATAGAGCAACTCATGCTGGCACAGCAGGGTTACACTCCAGATGACATCCTGCCAGA GGGTAAGAAAGTGAATTTGAGGCGGAACAGCAATATTAGTACTGGGGGAGACTCCATAGACGTCA CAGAAACAATGGATAGCTCTTATCAAGAACTTGCAGCAGCGATGGCTACCAGTATGGGGGCATGG GCCTGTGGAGTTGATCTGATTATACCCGACGAAACGCAGATTGCCACAAAGGAAAATCCACATTGC ACGTGTATTGAACTTAACTTCAACCCCTCCATGTACATGCATACATACTGCGCTGAGGGGCCGGG GCAGGCAATTACAACCAAAATACTCGACAAACTCTTCCCGGAGATCGTGGCCGGACAAACTGGAG GAAGCGGAGGAAGCGGAGGAAGCGATTACAAGGATGACGATGACAAGTAAGCTACGTAAATTCC GCCC Mito-GSHf GCCGGATCTAGCTAGTTAATTAAGccaccATGCTCGCTACAAGGGTCTTTAGCCTCGTCGGAAAGAG AGCTATCAGCACCTCCGTCTGCGTGAGAGCTCATatggcgccctacagcctactggtgactcggctgcagaaagctctg ggtgtgcggcagtaccatgtggcctcagtcctgtgcGATTACAAGGATGACGATGACAAGGGAGGAAGCGGAGGAA GCGGAGGAAGCACCCTTAATCAGCTTCTCCAGAAGTTGGAGGCGACTTCCCCCATTCTCCAGGCG AACTTCGGGATAGAAAGGGAGTCATTGAGGGTTGACCGCCAGGGTCAGCTGGTCCACACACCGC ACCCCTCATGTCTGGGAGCCCGCAGTTTTCATCCTTACATACAAACCGACTTTTGTGAATTCCAAAT GGAACTGATTACACCAGTAGCCAAAAGTACGACGGAGGCCCGACGCTTTCTTGGCGCGATAACTG ATGTAGCAGGACGAAGCATTGCAACTGACGAGGTGCTGTGGCCATTGAGTATGCCACCACGACTT AAAGCCGAGGAAATTCAAGTAGCGCAACTCGAGAACGACTTCGAAAGACATTATCGGAACTACTTG GCAGAGAAGTACGGCACCAAATTGCAGGCGATTAGTGGAATTCATTACAATATGGAACTTGGGAA GGACTTGGTTGAGGCGCTTTTTCAAGAGTCAGATCAGACTGACATGATCGCATTTAAAAACGCTCT GTATCTCAAGCTCGCCCAGAACTATTTGAGGTATCGGTGGGTCATTACTTACCTGTTTGGAGCAAG TCCCATTGCAGAACAAGGATTCTTTGACCAAGAAGTGCCGGAGCCTATGCGCTCTTTCCGCAACTC CGACCACGGCTACGTTAACAAGGAAGAGATACAGGTAAGCTTTGTATCCCTTGAAGACTATGTCTC CGCGATCGAGACCTACATCGAGCAGGGTGACCTTATAGCCGAGAAAGAGTTTTACTCAGCCGTGC GCTTTAGAGGACAAAAAGTCAATCGCTCCTTCCTTGATAAGGGTATAACTTATCTGGAGTTCAGAA ACTTTGACTTGAACCCATTTGAGAGAATAGGCATCAGTCAAACCACTATGGATACCGTTCACTTGC TCATACTGGCCTTTCTCTGGTTGGATAGTCCGGAAAACGTGGACCAGGCCCTTGCCCAGGGACAC GCGCTTAACGAGAAAATAGCCCTCTCCCATCCATTGGAGCCCCTCCCTTCAGAGGCCAAGACACA GGACATCGTGACCGCACTCGACCAGCTCGTACAGCACTTCGGATTGGGAGATTACCACCAGGATC TCGTGAAACAAGTGAAAGCGGCGTTTGCTGATCCGAATCAAACCCTGTCAGCTCAACTTCTTCCTT ATATTAAGGACAAGTCACTCGCAGAATTCGCTCTCAATAAGGCACTCGCATATCATGACTATGACT GGACCGCTCACTACGCCCTTAAAGGTTACGAAGAAATGGAGCTCAGTACGCAGATGCTGCTCTTT GATGCTATCCAGAAAGGAATACACTTCGAGATACTCGATGAGCAAGATCAGTTCTTGAAGCTGTGG CATCAAGATCACGTAGAATATGTTAAAAACGGTAATATGACCAGCAAGGATAACTATGTAGTACCTC TCGCAATGGCCAACAAGACTGTTACTAAGAAAATTCTTGCTGACGCTGGGTTCCCTGTTCCGTCCG GGGACGAATTTACTAGCTTGGAGGAGGGACTGGCCTACTACCCGCTTATTAAAGATAAGCAAATTG TAGTAAAGCCAAAGAGCACGAATTTCGGCTTGGGTATCAGCATCTTCCAAGAACCCGCCAGTCTC GACAATTATCAAAAAGCATTGGAAATAGCATTTGCGGAGGACACTAGTGTGCTCGTCGAAGAATTC ATTCCAGGCACGGAATACCGATTCTTCATTTTGGACGGACGCTGTGAGGCAGTCCTTTTGAGGGT AGCTGCCAATGTAATAGGGGACGGGAAACACACAATCAGAGAGTTGGTAGCGCAGAAAAACGCAA ATCCCCTGCGCGGTAGGGATCATAGATCACCCTTGGAAATCATAGAGCTTGGGGACATAGAGCAA CTCATGCTGGCACAGCAGGGTTACACTCCAGATGACATCCTGCCAGAGGGTAAGAAAGTGAATTT GAGGCGGAACAGCAATATTAGTACTGGGGGAGACTCCATAGACGTCACAGAAACAATGGATAGCT CTTATCAAGAACTTGCAGCAGCGATGGCTACCAGTATGGGGGCATGGGCCTGTGGAGTTGATCTG ATTATACCCGACGAAACGCAGATTGCCACAAAGGAAAATCCACATTGCACGTGTATTGAACTTAAC TTCAACCCCTCCATGTACATGCATACATACTGCGCTGAGGGGCCGGGGCAGGCAATTACAACCAA AATACTCGACAAACTCTTCCCGGAGATCGTGGCCGGACAAACTTAAGCTACGTAAATTCCGCCC Human codon-optimized Cyto-GshF was custom synthesized by IDT and directly assembled into pMXS-puro linearized with BamHI and NotI with a Gibson reaction
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