Appendix 2: Genetic Sequences

Electronic versions of the sequences are available on Canvas.

CA Human carbonic anhydrase II thermostable variant gene

Gene with available TAG sites (bold and underlined- Phe20, Phe93, Trp97, Lys126, Glu186, and Glu233), 6his-tag (lowercase), and thermostable mutations (lowercase)

 ATGGCGCATCATTGGGGTTACGGTAAACACAACGGTCCGGAGCATTGGCAC-

AAAGATTTTCCAATTGCGAAGGGCGAACGTCAAAGCCCGGTTGACATTGAT-

ACGCACACGGCAAAGTACGACCCGAGCCTGAAACCGCTGAGCGTTTCCTAT-

GACCAGGCTACGAGCCTGCGTATCCTGAACAATGGCCACaccTTCAACGTGG-

AGTTTGATGATTCCCAAGATAAGGCGGTTCTGAAAGGTGGTCCGTTGGATG-

GCACCTACCGCCTGATCCAATTTCACTTTCACTGGGGTAGCcacGACGGTCA-

GGGCAGCGAGCATACCGTGGACAAAAAGAAGTATGCAGCCGAACTGCACCT-

GGTGCATTGGAACACGAAGTACGGCGACTTCGGTAAAGCGGTCCAGCAACC-

GGACGGTCTGGCTGTTCTGGGTATTTTCCTGAAGGTCGGCAGCGCGaacCCG-

GGTCTGCAGAAAGTGGTTGACGTGTTGGACTCTATCAAGACCAAAGGCAAGA-

GCGCGGACTTCACCAATTTCGATCCGCGTGGTCTGCTGCCGGAGAGCCTGGA-

TTACTGGACTTATCCGGGCAGCCTGACCACCCCGCCATTGCTGGAGTGCGTG-

ACCTGGATCGTCTTGAAAGAACCGATCAGCGTTAGCTCTGAACAGGTCagcAA-

GTTCCGCAAGCTGAATTTCAATGGTGAGGGCGAGCCGGAAGAAccgATGGTC-

GATAATTGGCGTCCTaccCAACCGCTGAAAAACCGCCAGATTAAAGCATCCTTTA-

AGcatcaccaccaccatcactaa
  Mutations to increase thermostability A65T, L100H, K153N, L223S, L239P, A247T

 

HPII E.coli catalase HPII gene

Gene with 6his-tag and available TAG sites (bold and underlined- Phe206, Glu283, Lys348, His392, Tyr415, and Asp568) and 6his-tag with linker (lowercase)

 

 atggggggttctcatcatcatcatcatcatggtatggctagcatgactggtggacagcaaatgggtcggga-

tctgtacgacgatgacgataaggatcgatggggatccgagctcgagTCGCAACATAACGAAAAG-

AACCCACATCAGCACCAGTCACCACTACACGATTCCAGCGAAGCGAAACCGG-

GGATGGACTCACTGGCACCTGAGGACGGCTCTCATCGTCCAGCGGCTGAACC-

AACACCGCCAGGTGCACAACCTACCGCCCCAGGGAGCCTGAAAGCCCCTGAT-

ACGCGTAACGAAAAACTTAATTCTCTGGAAGACGTACGCAAAGGCAGTGAAAA-

TTATGCGCTGACCACTAATCAGGGCGTGCGCATCGCCGACGATCAAAACTCAC-

TGCGTGCCGGTAGCCGTGGTCCAACGCTGCTGGAAGATTTTATTCTGCGCGAG-

AAAATCACCCACTTTGACCATGAGCGCATTCCGGAACGTATTGTTCATGCACGC-

GGATCAGCCGCTCACGGTTATTTCCAGCCATATAAAAGCTTAAGCGATATTACC-

AAAGCGGATTTCCTCTCAGATCCGAACAAAATCACCCCAGTATTTGTACGTTTCT-

CTACCGTTCAGGGTGGTGCTGGCTCTGCTGATACCGTGCGTGATATCCGTGGC-

TTTGCCACCAAGTTCTATACCGAAGAGGGTATTTTTGACCTCGTTGGCAATAACA-

CGCCAATCTTCTTTATCCAGGATGCGCATAAATTCCCCGATTTTGTTCATGCGGT-

AAAACCAGAACCGCACTGGGCAATTCCACAAGGGCAAAGTGCCCACGATACTTTC-

TGGGATTATGTTTCTCTGCAACCTGAAACTCTGCACAACGTGATGTGGGCGATGT-

CGGATCGCGGCATCCCCCGCAGTTACCGCACCATGGAAGGCTTCGGTATTCACAC-

CTTCCGCCTGATTAATGCCGAAGGGAAGGCAACGTTTGTACGTTTCCACTGGAAAC-

CACTGGCAGGTAAAGCCTCACTCGTTTGGGATGAAGCACAAAAACTCACCGGACGT-

GACCCGGACTTCCACCGCCGCGAGTTGTGGGAAGCCATTGAAGCAGGCGATTTTCC-

GGAATACGAACTGGGCTTCCAGTTGATTCCTGAAGAAGATGAATTCAAGTTCGACTT-

CGATCTTCTCGATCCAACCAAACTTATCCCGGAAGAACTGGTGCCCGTTCAGCGTGT-

CGGCAAAATGGTGCTCAATCGCAACCCGGATAACTTCTTTGCTGAAAACGAACAGGC-

GGCTTTCCATCCTGGGCATATCGTGCCGGGACTGGACTTCACCAACGATCCGCTGTT-

GCAGGGACGTTTGTTCTCCTATACCGATACACAAATCAGTCGTCTTGGTGGGCCGAAT-

TTCCATGAGATTCCGATTAACCGTCCGACCTGCCCTTACCATAATTTCCAGCGTGAC-

GGCATGCATCGCATGGGGATCGACACTAACCCGGCGAATTACGAACCGAACTCGATT-

AACGATAACTGGCCGCGCGAAACACCGCCGGGGCCGAAACGCGGCGGTTTTGAATC-

ATACCAGGAGCGCGTGGAAGGCAATAAAGTTCGCGAGCGCAGCCCATCGTTTGGCGA-

ATATTATTCCCATCCGCGTCTGTTCTGGCTAAGTCAGACGCCATTTGAGCAGCGCCATA-

TTGTCGATGGTTTCAGTTTTGAGTTAAGCAAAGTCGTTCGTCCGTATATTCGTGAGCG-

CGTTGTTGACCAGCTGGCGCATATTGATCTCACTCTGGCCCAGGCGGTGGCGAAAAAT-

CTCGGTATCGAACTGACTGACGACCAGCTGAATATCACCCCACCTCCGGACGTCAACG-

GTCTGAAAAAGGATCCATCCTTAAGTTTGTACGCCATTCCTGACGGTGATGTGAAAGGT-

CGCGTGGTAGCGATTTTACTTAATGATGAAGTGAGATCGGCAGACCTTCTGGCCATTCT-

CAAGGCGCTGAAGGCCAAAGGCGTTCATGCCAAACTGCTCTACTCCCGAATGGGTGAA-

GTGACTGCGGATGACGGTACGGTGTTGCCTATAGCCGCTACCTTTGCCGGTGCACCTT-

CGCTGACGGTCGATGCGGTCATTGTCCCTTGCGGCAATATCGCGGATATCGCTGACAA-

CGGCGATGCCAACTACTACCTGATGGAAGCCTACAAACACCTTAAACCGATTGCGCTG-

GCGGGTGACGCGCGCAAGTTTAAAGCAACAATCAAGATCGCTGACCAGGGTGAAGAAG-

GGATTGTGGAAGCTGACAGCGCTGACGGTAGTTTTATGGATGAACTGCTAACGCTGATG-

GCAGCACACCGCGTGTGGTCACGCATTCCTAAGATTGACAAAATTCCTGCCTGA

 

 

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Chemical Biology & Biochemistry Laboratory Using Genetic Code Expansion Manual Copyright © 2019 by Ryan Mehl, Kari van Zee & Kelsey Keen is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License, except where otherwise noted.