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Parts overview

Name

Role

Type

P30_PL28
Endo-acting ulvan lyase

This gene codes for an enzyme which catalyze rapid dissolution of insoluble ulvan: this enzyme cleaves the glycosidic linkage between Rha3S and GlcA or IduA, producing oligosaccharides with unsaturated uronic acid at the non-reducing end.

Coding

Codon-optimized for E.coli (TWIST) nucleotide sequence

ATGTTCCTGGCAGTGGTGACCGCTCAGACTGCACCTGATGAAGACACGTCCGCGATCACGCGCTGCACGGCAGAGGGCACTAATCCGGTGCGCGAGACCG
ACATCCCGAATCCGGTGAACGTAGGTACGATTGACGATCGCTCATGCTACGCCAACTACAAAGAATCTACCGTATACGGCAAGACATGGGGCGTTTACAA
TATTACCTTCGACTCGAACGACTTCGACACGTCGCTCCAGCCGCGCATCGAGCGCAGTCTGAGTCGTTCATCAGAGACCGGCATCGGCTCATACGCACGC
CTTACCGGTGTATTCCGTATCCTGGAAGTGGGTGACACGAGCGGAACTTCGCAAGACGGGACCTACCTGGCACAGGCAAAGGGTAAACATACGGGCGGTG
GCGGCTCTCCGGACCCGGCGATCTGCCTGTACCTCGCAAAGCCGGTGTACGGCACTGGCGAGGATGCAGACAAGCAAGTTAGTTTTGACATCTACGCGGA
ACGCATCCTTTACCGCGGCGGCGAGGGTGACGGGCGCGAGATCGTGTTTCTTAAGAACGTTAAGAAAGACGAGGAGACAAACTTCGAACTTGAGGTCGGC
TTTAAAGAGGACCCGAATGACGTTAGCAAGAAGATTCAATACTGCAACGCAGTGATCGGCGGTGACACCTTCAACTGGAACATCCCGGAGCCCGAACGCG
GCACCGAAAGTGGCATCCGCTACGGCGCGTACCGTGTAAAGGGCGGTCGTGCCCAAATTCGTTGGGCAAACACGACTTATCAGAAAGTGGAGAATGTCGA
AGTCACCAACCCGGGCCCTATTGGCGACGTGTACAAGCTGAAGAACGTGGCCACCGGGCAATACCTGAGCGACAGTGGCGTATCCGCGTCGGCGGTGATC
ATGAGCGACAGTGGGGAAGCCCAGAACAACTACTGGACATTCGTTGAGTCAGGTTCTTTATTCAATATTGACAACGAAACATTTGGAATCCTTCGCGCGC
CTGGCGCGGGCGGACCGGGCGGGGCATACGTTGTGGTTAGCACCACCAAGGAAGGGCCGTCCTCAGACGGAGACAAAGTATGGACCATTCACTACAACGA
GTCAAATGACACCTACCGTTTCGAGTCAGGCAGCTCTGGGCGTTTCATGTACCAGGAGATCAACGGTAACGTGACCCACATTAGCGCCATGAACACGGAC
GACCGTTCTGTGTGGAAGGCGATCGCGGTCGAATCGCTTAGCGTCGACGAAAATGCAATCCTGGCAAGCGACGTGCGCGTTTTCCCAAATCCCGCGAGTG
ACAGCTTCACCATCTCACTGAAGACCATCAACCACGTAACGGTGAACATTTATGACGTCCTTGGTAACACCATCTTCAAGTCTGAGTTCAACGGCGACAC
GATCCAGATCCGTAACAAGGGACAGTTTAAGGCCGGCGTATACTTAATCCAACTCACGGACAAGAACAATAATAAATACCACAAGAAGCTGATTGTGAAG
GGTGCTGGTCATCATCACCATCACCACGAGAACCTGTACTTCCAATCCAATTTAGCCTATGGAAGTCAGGGTGATCTTAATCCATTAATTAATGAAATCA
GCAAAATCATTTCAGCAGCAGGTAGCTTCGATGTTAAAGAGGAAAGAACCGCAGCTTCTTTATTGCAGTTGTCCGGTAATGCCAGTGATTTTTCATATGG
ACGGAACTCAATAACCCTGACCACATCAGCATAATA

•  RED : Linker
•  GREEN : His Tag sequence
•  BLUE : TEV cleavage sequence
•  YELLOW : alpha-haemolysin signal sequence (hylA)

Protein sequence

MFLAVVTAQTAPDEDTSAITRCTAEGTNPVRETDIPNPVNVGTIDDRSCYANYKESTVYGKTWGVYNITFDSNDFDTSLQPRIERSLSRSSETGIGSYAR
LTGVFRILEVGDTSGTSQDGTYLAQAKGKHTGGGGSPDPAICLYLAKPVYGTGEDADKQVSFDIYAERILYRGGEGDGREIVFLKNVKKDEETNFELEVG
FKEDPNDVSKKIQYCNAVIGGDTFNWNIPEPERGTESGIRYGAYRVKGGRAQIRWANTTYQKVENVEVTNPGPIGDVYKLKNVATGQYLSDSGVSASAVI
MSDSGEAQNNYWTFVESGSLFNIDNETFGILRAPGAGGPGGAYVVVSTTKEGPSSDGDKVWTIHYNESNDTYRFESGSSGRFMYQEINGNVTHISAMNTD
DRSVWKAIAVESLSVDENAILASDVRVFPNPASDSFTISLKTINHVTVNIYDVLGNTIFKSEFNGDTIQIRNKGQFKAGVYLIQLTDKNNNKYHKKLIVK
GAGHHHHHHENLYFQSNLAYGSQGDLNPLINEISKIISAAGSFDVKEERTAASLLQLSGNASDFSYGRNSITLTTSA—

•  RED : Linker
•  GREEN : His Tag sequence
•  BLUE : TEV cleavage sequence
•  YELLOW : alpha-haemolysin signal sequence (hylA)

Name

Role

Type

P10_PLnc
Endo-acting ulvan lyase

Enzyme coded by this gene dissolves soluble ulvan oligomers at the cell surface. The localisation of this protein is not sure for now between outer membrane or periplasmic. This enzyme cleaves also the glycosidic linkage between Rha3S and GlcA or IduA, producing oligosaccharides with unsaturated uronic acid at the non-reducing end.

Coding

Codon-optimized for E.coli (TWIST) nucleotide sequence

ATGAAAACCAAAAACTTAATTCTTTTAGTTGGTATTCTTTTCAACATGTCCGCTGTGGCTAGTGAAACGTATCACAATATAATTTTAAGTAGTACACAGC
AGTCGAAAGCAGCGAAGGTTAATGAAACCATTACATATACATTTGAAATTAATAACGTTGATGATACGGAACAAGTATATCATATTCAAATTATAAACCC
GAGGACTTTAGCATGTCAGGTTACTTTAAAAGATACTTTAGTTACTGTTAAAAGCAATCAAGTGTATCACGGAGCGTTTTTAGTCCGAGTTAGTGATGGA
ATACCTGTTGGCGGACAAGAAACAATTTTTTTAGATATAAAAAATGAAGCGAATACACGTATTGAAACTTTAGAATTTATTACGGTACGGTATAAACCAC
ATCCATTTCTTTTGGTAACCGATGAGATTTTAGAGGAGGCTAAAGAAAAGGTAAAAAATGTAGATTGGGCAAAGCAAAATCTAGAAGAACTTATCGCTTT
TGCGAATGCTTATTACGTTCCAGAACGTAAGGTGGTAGATAAACCAAGACCGGTTAAGCGTTGGAAAAGTTTAGCTTATAAAGCAGCAGATGGTGAAGAT
CTTTTTAAAGTAGCACTTGCCTATAAATTTACAGGAAATACAACGTATTTAAAAAAAGTGATCCAGCTTGTTAATGATGTTTGCGACCCGGAAGAAGGCT
ATCTGTCTCTAGGTGCCGCAACTACAGGCGTTCAGGTCCATGAAGGATTCTTTTTTATGTATTTGGCATTAACCTGCGATATTTTATATGGTGAAGATGT
CTTGTCTGATATAGAACGTCAACATATTGAACAATCCATGCGTCTGTACTTAAAACAAGACAAATCTCATATGGATCCTATCGGGATTATGAACCATCAA
GCGAGTGCCAATGTTGGGGCTGTAATCACGTCTCTTTTTCTTCAAGATATGACTGCAGTGAATCATTTTGTAAAAAGCGATGGCGGACTTATAGATCAAA
TGACCAAAGGGATTATGGCAGATGGTTGGTGGTTTGAAGGTACTCCTGGCTATTGTTATTTGGTAACAGAAAGATTTTGTCTTACTGCTCAAGCTTTCGA
AAATTACGGATGGAATTTATACGACAGACGTTTTCCTATGCGATTTAAAAGTAAAGATTTCGAAAACGCCAAAGAAGGTTACACGGGAATGAAGTTCGAT
ATTTGGGGCCCTACAGGTAGCAGTACCAGAGGACTTAAAGAGATGGTGAACGCTTATATTCCTATGATGGATGAAGAGGCCAATTTAATTCCTAATAACG
ACGGAACCGTTTCAAAACCTCATGAGTTTTATGAGTTGGCTTACAGACATTTTAAAAATGACGATTTAGCATGGGTAATCAGCAAATCTAAAAGAGACAG
TTGGGAATCGTTATTATATGGTGTTGCAGAACTTCCGGAAGTAAAAGACCCTAGATCAGAATCGGAATATGTGTCTAATGTTGGTTTGGTAGCCTTGCGT
TCTCAACCAAAAAATAAAATCCCAGAAGAACAAATTCAAGCCTATTATAAATTTGGAACACATGGCGGATGGCACGGGCATTTAGATAGAACCGGAATGA
TTGCTTTAAACCGTTACGGGCATAAGTATTTTGGAACCGAAATGGTGTGGTTTGGCTATGGCGATCCGGGGTATAAAGAGAGTGTTCAAGCATCGGCAAC
ACACAATATGGTTGTTATTGATGGTTTACAACAAGAAGCTGTACCATCAGAACAACTATTGTTTTATAAAGGCGATATCATGCAAGCCACTGTGGTGCAA
ACAAAAGCCCGATGGAGAAGAATTCCCATTTTTAATAAAGACCTATTTCCACCATGGAATGATTTCGATATTGAACCTGGTTTTGAACCTGTATTACAAA
GACGTTTGTCTGTGGTAACAACTGATTATGTTGTGATTGTTGACTATATGCAGGCGCCTCAAAAACATACTTACGATTGGATGTTACATCCTGTTGGATT
TCAGTCTATAGATGGTGTCACAACAAAAGGCGGTGTCAAACCTACGTGGAGTGAGGATTTAAATTCTCAATATAAATACTTTACAAGCGCACAATGGTAT
ACATCAAATAAAACGAAAAAAGTCAAGTTTCAAGAACACGATATGAAGTTAGATGTCCATACCCTTTGGCCAAAAAAAGCGGATGTTGTTGTGGCGAGTT
ATCCCAATGGTGGTAAGCAACGAGGCATTAGGAACAATCCAAATCGCGCAACTTATGGCGTTCGAGTATCGGGTAAAGAAGCGCAGTTTTTAACGGTTCT
AGAACCATATAAAAACGCGTCTGCAATTTCTAAGATAGTATCGAAAAATGAAAATGAACTCACAGTGTTTCTGAAAGATGGAAGGAAGCAAAGTGTTTTT
ATAGAGCATATTAAAGATGCAAATATAAAAATCACTATTAAAGAGTATGTGAATGATGAGTTAGTTGCTACAAATCAGGCTCTAGCGACAACGAACGTGC
TGGTTACCCATACGATGTTCCAGATTACGCTGAGAACCTGTACTTCCAATCCAATTTAGCCTATGGAAGTCAGGGTGATCTTAATCCATTAATTAATGAA
ATCAGCAAAATCATTTCAGCAGCAGGTAGCTTCGATGTTAAAGAGGAAAGAACCGCAGCTTCTTTATTGCAGTTGTCCGGTAATGCCAGTGATTTTTCAT
ATGGACGGAACTCAATAACCCTGACCACATCAGCATAATAA

•  RED : Linker
•  GREEN : His Tag sequence
•  BLUE : TEV cleavage sequence
•  YELLOW : alpha-haemolysin signal sequence (hylA)

Protein sequence

MKTKNLILLVGILFNMSAVASETYHNIILSSTQQSKAAKVNETITYTFEINNVDDTEQVYHIQIINPRTLACQVTLKDTLVTVKSNQVYHGAFLVRVSDG
IPVGGQETIFLDIKNEANTRIETLEFITVRYKPHPFLLVTDEILEEAKEKVKNVDWAKQNLEELIAFANAYYVPERKVVDKPRPVKRWKSLAYKAADGED
LFKVALAYKFTGNTTYLKKVIQLVNDVCDPEEGYLSLGAATTGVQVHEGFFFMYLALTCDILYGEDVLSDIERQHIEQSMRLYLKQDKSHMDPIGIMNHQ
ASANVGAVITSLFLQDMTAVNHFVKSDGGLIDQMTKGIMADGWWFEGTPGYCYLVTERFCLTAQAFENYGWNLYDRRFPMRFKSKDFENAKEGYTGMKFD
IWGPTGSSTRGLKEMVNAYIPMMDEEANLIPNNDGTVSKPHEFYELAYRHFKNDDLAWVISKSKRDSWESLLYGVAELPEVKDPRSESEYVSNVGLVALR
SQPKNKIPEEQIQAYYKFGTHGGWHGHLDRTGMIALNRYGHKYFGTEMVWFGYGDPGYKESVQASATHNMVVIDGLQQEAVPSEQLLFYKGDIMQATVVQ
TKARWRRIPIFNKDLFPPWNDFDIEPGFEPVLQRRLSVVTTDYVVIVDYMQAPQKHTYDWMLHPVGFQSIDGVTTKGGVKPTWSEDLNSQYKYFTSAQWY
TSNKTKKVKFQEHDMKLDVHTLWPKKADVVVASYPNGGKQRGIRNNPNRATYGVRVSGKEAQFLTVLEPYKNASAISKIVSKNENELTVFLKDGRKQSVF
IEHIKDANIKITIKEYVNDELVATNQALATTNGAGYPYDVPDYAENLYFQSNLAYGSQGDLNPLINEISKIISAAGSFDVKEERTAASLLQLSGNASDFS
YGRNSITLTTSA—

•  RED : Linker
•  GREEN : His Tag sequence
•  BLUE : TEV cleavage sequence
•  YELLOW : alpha-haemolysin signal sequence (hylA)

Name

Role

Type

P33_GH105
Unsaturated 3S-rhamnoglycuronyl hydrolase

Enzyme coded by this gene is periplasmic and works together with ulvan lyases to fully degrade the ulvan polymer. It catalyzes specifically the cleavage of the unsaturated uronic acid at the non-reduced end of the oligosaccharides generated by the ulvan lyase.

Coding

Codon-optimized for E.coli (TWIST) nucleotide sequence

ATGAAGAATCAGGCTCTGAAGATCTTAACCCTTTGCGTCCTCGTAGGCAGTGCTATGAGCTTAAAGTTGTACGCCCAGAAGGGCCTGAATCACAGTGAGA
TCGAGGCAAAGATGATCAAAGCACTGGAGTGGCAGGAAGCTCATCCGATCTTTGCCCTGGCTCCGACCGACTGGACGGAAGGGGCCTACTATATCGGCGT
GTCTCGCGCCCACAAAACAACGCAAGACATGATGTACATGGCGGCTCTGAAGAATCAGGCCTACTGGAACAACTGGCAGACGTACTCGCGTTTGCACCAT
GCAGACGACGTTGCGATTAGTTACTCTTACATCTACATCGGAATGAATGACAAGCGCCCGGGTTTCGTTAATTTGGAACCAACCAAGAAATTCCTGGACG
CGCATCTGCATGAAGACGACGAGTGGAAAGCAGGCACGGACAAGTCCGCAAGCGGCAAGACAATCTTGTGGTGGTGGTGTGACGCGCTGTTCATGGCACC
GCCGGTTCTGAACTTGTACGCCAAACACACCAATCAACCGAAATACCGTGACGAGATGCACAAATACTATATGGAGACGTACAACCAACTTTATGACAAG
GAAGAGCGCCTGTTCGCCCGCGACATGCGCTTCGTGTGGAAAGGCACCGAGAAGGACCTGAAGGAGCCTAACGACAAGAAGATTTTCTGGAGCCGCGGTA
ACGGCTGGGTACTGGGCGGCCTGGCCCTCTTGCTGGACGACATGCCGAACGACTACAAGCATCGTACTTTCTACGAGAACTTGTTCAAAGACATGGCCAG
TCGTATCCTGGAGTTGCAGCCAAAGGACGGACTGTGGCGCACCTCGCTTTTGTCCCCCGAGACTTACGACCACGGCGAGGTTTCCGGCTCCGGCTTTTAT
ACTTTCGCACTGGCGTGGGGCGTGAACAACGGGCTGCTGGATCGTAACAAGTACGAGCCTGCGGTTAAGAAGGCTTGGAAAGCGCTGGCAGATTGTCAAC
ACGAGGATGGCCGCGTCGGCTGGGTACAGAACATCGGTGCAAGCCCAGAGCCGGCGTCAGCCGACTCATGGCAGAACTTCGGTACCGGGGCTTTCCTCAT
GGCGGGAAGCGAGGTGCTGAAGTTAGAGGAAGGTGCTGGT GACTACAAAGACGATGACGACAAGGAGAACCTGTACTTCCAATCCAATTTAGCCTATGGA
AGTCAGGGTGATCTTAATCCATTAATTAATGAAATCAGCAAAATCATTTCAGCAGCAGGTAGCTTCGATGTTAAAGAGGAAAGAACCGCAGCTTCTTTAT
TGCAGTTGTCCGGTAATGCCAGTGATTTTTCATATGGACGGAACTCAATAACCCTGACCACATCAGCATAATAA

•  RED : Linker
•  GREEN : His Tag sequence
•  BLUE : TEV cleavage sequence
•  YELLOW : alpha-haemolysin signal sequence (hylA)

Protein sequence

MKNQALKILTLCVLVGSAMSLKLYAQKGLNHSEIEAKMIKALEWQEAHPIFALAPTDWTEGAYYIGVSRAHKTTQDMMYMAALKNQAYWNNWQTYSRLHH
ADDVAISYSYIYIGMNDKRPGFVNLEPTKKFLDAHLHEDDEWKAGTDKSASGKTILWWWCDALFMAPPVLNLYAKHTNQPKYRDEMHKYYMETYNQLYDK
EERLFARDMRFVWKGTEKDLKEPNDKKIFWSRGNGWVLGGLALLLDDMPNDYKHRTFYENLFKDMASRILELQPKDGLWRTSLLSPETYDHGEVSGSGFY
TFALAWGVNNGLLDRNKYEPAVKKAWKALADCQHEDGRVGWVQNIGASPEPASADSWQNFGTGAFLMAGSEVLKLEEGAGDYKDDDDKENLYFQSNLAYG
SQGDLNPLINEISKIISAAGSFDVKEERTAASLLQLSGNASDFSYGRNSITLTTSA—

•  RED : Linker
•  GREEN : His Tag sequence
•  BLUE : TEV cleavage sequence
•  YELLOW : alpha-haemolysin signal sequence (hylA)

Name

Role

Type

P36_ S1_25
Bifunctional sulfatase/alpha-L-rhamnosidase

Enzyme coded by this gene is periplasmic and is a bifunctional enzyme containing sulfatase and alpha-L-rhamnosidase activities involved in ulvan degradation. The sulfatase is the part of this protein that interests us. It specifically desulfates L-rhamnose at the 3-position. The architecture of this protein with 2 functions is very rare.

Coding

Codon-optimized for E.coli (TWIST) nucleotide sequence

ATGAAGAATCAGGCTCTGAAGATCTTAACCCTTTGCGTCCTCGTAGGCAGTGCTATGAGCTTAAAGTTGTACGCCCAGAAGGGCCTGAATCACAGTGAGA
ATGAGTCAGACGGTGAAGAAGGAAAAGCCGAACATTATCTTTATCTTGACCGATGACCAACGCTTCGACGCAATTGGGTACGCCGGCAATAAGTTCGTCA
ACACGCCGGAGATGGATAAGCTGGCGCAGCAGGGCACTTACTTCGACCACGCCATCGTGACGACCCCAATCTGCGCGGCTAGCCGCGCAAGTCTGTGGAC
AGGCCTGCATGAACGCAGCCACAACTTCAATTTCCAGACTGGCAACGTGCGTGAGGAGTATATGAACAACGCGTACCCGAAGCTGCTGAAGAACAACGGT
TACTACACGGGCTTCTACGGCAAGTACGGGGTCCGCTACGACAATTTAGAAAGCCAGTTCGACGAGTTCGAATCATACGATCGCAACAACCGTTACAAGG
ATAAGCGCGGATACTATTACAAAACCATCAACAATGACACCGTTCACCTTACTCGCTACACGGGCCAGCAGGCAATTGATTTCATTGACAAGAACGCCAC
GAATACGCAGCCCTTCATGCTTAGCCTGTCGTTCTCGGCCCCTCACGCGCACGACGGTGCGCCGGAACAGTACTTCTGGCAGACGACCACGGACGCATTA
TTGCAGGACACCACCCTGCCGGGGCCTGATCTGGCCGATGAGAAGTATTTCCTGGCTCAGCCTCAGGCAGTCCGCGACGGTTTCAACCGCCTGCGCTGGA
CCTGGCGCTATGATGATCCGGAGAAGTACCAGCATTCTCTGAAGGGTTACTATCGCATGATCAGTGGTATTGACCTGGAGATTAAGAAGATCCGCGACAA
GCTGAAGGAGAAAGGGGTCGATAAGAATACAGTTATCATCGTCATGGGCGATAACGGATATTTCCTGGGCGAGCGTCAGCTCGCGGGCAAGTGGCTCATG
TATGACAATTCCATCCGTGTCCCGCTCATCGTCTTCGACCCACGCGTCAACAAGCACCAGGACATCAGCGAGATGGTCCTCAACATTGACGTGACGCAGA
CTATTGCCGACCTGGCCGGTGTGAAGGCGCCTGAGTCCTGGCAGGGCAAGTCGCTTTTGCCACTTGTGAAACAGGAAACTTCTACTATCAGTCGCGATAC
GATCCTGATTGAACACCTTTGGGACTTCGAGAACATTCCACCCAGCGAGGGCGTGCGCACCGAGGAGTGGAAGTATTTCCGCTACGTGAACGATAAGACT
ATTGAGGAACTGTACAATATTAAGAAGGACCCGAAGGAGATCAACAACCTTATTGGCAAGAAGAAGTATCAAAACGTAGCCAAAGCGCTGCGTGAGAAGT
TGGACGAACTTATTGCGAAGAACTCTGACGAGTTCCGTGCGGGTCCTTCCGACCTGACGGTAGAGTTGATCCGCCAACCGGAGAGCGAGGTCAAGATCTT
TGACCTGAAGCCGGAGTTCGGCTGGACTGTCCCCTTAAGTAGCAAGTATCAGTCCGCGTACCAACTGCTGGTAGCGAGCTCAGAGACCATTATCAACGCG
AACAACGGTGACGTGTGGGACAGCGGGCAGGTTCGTTCGAGCCAGAGCACAAACGTAGACTTCGGCGGCAAGCCGTTGAAGATTGGAGAGACCTATTACT
GGAAGGTTCGCATCTGGGACGAGGAGAATCGTCTTGTTGATTACAGTAAGGCCCAGAAGTTTACGATCGGAGAGAGCGACAATTACATCATCTCCACTGA
GAACAAGTTCGTGACTGACAAAATCAAACCGAGCAAGTTCGAAAACCGCGACGGAGTTTACTTCATTGACTTTGGTAAGGCGGCCTTCGCGACGATGGAA
TTTAACTACCAGGCTAAGACCCCGCACACGCTGACCATCCGTGTGGGCGAGATGATTGACGAAAATGGAAATGTTAATCGCACGCCGCCCGCGAAGTCGA
ATATCCGCTACCAAGAACTTAAAGTCGAAGTAAAGCCGGGTCAGACCCGCTACCGTATCCCGATCCAGACCGATGAACGTAACACGCGTCCGAATAAAGC
CATCCCCCTGCCGAAGGGTTTCCCGCCCCTGCTGCCGTTCCGTTACGCTGAGATCGAGGGTGCGCAGAGTAGCATCAACGCAAATGACGTCGAGCAGCTT
GCTTACCACACCTTCTGGGATGAGAAGGCCTCATCGTTCAAGTCCGACAACAACATCCTCAACCAGGTGTGGGACCTTTCCAAGTACTCAATCAAGGCGA
CGACGTTCAACGGCTTATACGTCGACGGGGACCGTGAGCGCATCCCGTATGAGGCGGACGCATACTTAAACCAGCTGTCCCACTACACCACCGATCGCGA
GTACGCTATGGCGCGTCGCACCATTGAATACTTTATGAAGAATCCGACGTGGCCGACTGAGTGGCAACAGCACGTGGCCTTATTGCTGTACGCCGACTAC
ATGTACACGGGTAACACGGAACTTGTCGAACGCTACTACGAGGCGCTGAAGCACAAGAGCTTATATGAACTGAGCAACGAGGACGGTCTGATCACCAGCA
CCAAGGTGGACGCGGAATTTATGAAGAAGCTTGGGTTTCCTGAGGGCTATAAGAAACCGTTGACGGACATCGTGGATTGGCCTGGCGCGAACTTTAACGG
TTCGAAGACTCCGGGTGAGCGCGATGGGTTCGTTTTCCAACCGTACAACACCGTCATCAACAGTTTCTTTTATGAAAACATGAAGATTATGGCACAGTTC
GCTAAGATTCTCGGAAAGACGGACGAAGTGCTGGACTTTGAGCTGCGTGCGGCAAAGGCCAAGAAGGCCGTTAATGAACAGATGTTTGACAAGAAGCGCG
GCATCTACGTAGACGGAATCGGCACGGACCACGCCAGCCTGCACGCAAACATGATGCCGCTGGCTTTCGGCCTGGTGCCGCAGGAGCACGTGGACACCGT
CGTGGAGTTCGTTAAGTCCCGCGGGATGGCATGCTCTGTGTACGGCGCTCAGTTTCTTCTGGACGGCCTTTATAACGTTGGCGAGGCCGACTACGCTCTG
GACTTGCTCGCCAGCACTTCTGAGCGCTCATGGTACAACATGATCCGCATCGGTAGCACGATCACCTTAGAGGCCTGGGACAACAAATACAAGAATAACC
TGGACTGGAACCACGCATGGGGTGCCGTTCCGGCGAACGCCATTCCCCGTGGTCTGTGGGGTATCAAGCCGAAAACCGCGGGCTTCGGCATTGCGAGCAT
TAAGCCGCAGATGGGGAAGCTGAAATCGTCACAGATCACAGTCCCCACTGTGCGCGGAGCGATTCACGCGACGTTCACGCACAACGGCCCGCGCTCTCAG
ACCTACGAGATCGAGATCCCGGGCAATATGGTTGCGGAGTTCTCCCTGGACGATATTGACGGCAAAGACCTGATCCACAATGGCCAAAAGGTTCCTGCCG
CCTTCGGCGCAGTTCAGCTGTCCCCTGGAAAGCACATCATCGAGCTGAAGATCAATTCTTTCGGTGCTGGTTTGGAGCCACCCGCAGTTCGAAAAAGAGA
ACCTGTACTTCCAATCCAATTTAGCCTATGGAAGTCAGGGTGATCTTAATCCATTAATTAATGAAATCAGCAAAATCATTTCAGCAGCAGGTAGCTTCGA
TGTTAAAGAGGAAAGAACCGCAGCTTCTTTATTGCAGTTGTCCG

•  RED : Linker
•  GREEN : His Tag sequence
•  BLUE : TEV cleavage sequence
•  YELLOW : alpha-haemolysin signal sequence (hylA)

Protein sequence

MSQTVKKEKPNIIFILTDDQRFDAIGYAGNKFVNTPEMDKLAQQGTYFDHAIVTTPICAASRASLWTGLHERSHNFNFQTGNVREEYMNNAYPKLLKNNG
YYTGFYGKYGVRYDNLESQFDEFESYDRNNRYKDKRGYYYKTINNDTVHLTRYTGQQAIDFIDKNATNTQPFMLSLSFSAPHAHDGAPEQYFWQTTTDAL LQDTTLPGPDLADEKYFLAQPQAVRDGFNRLRWTWRYDDPEKYQHSLKGYYRMISGIDLEIKKIRDKLKEKGVDKNTVIIVMGDNGYFLGERQLAGKWLM YDNSIRVPLIVFDPRVNKHQDISEMVLNIDVTQTIADLAGVKAPESWQGKSLLPLVKQETSTISRDTILIEHLWDFENIPPSEGVRTEEWKYFRYVNDKT IEELYNIKKDPKEINNLIGKKKYQNVAKALREKLDELIAKNSDEFRAGPSDLTVELIRQPESEVKIFDLKPEFGWTVPLSSKYQSAYQLLVASSETIINA NNGDVWDSGQVRSSQSTNVDFGGKPLKIGETYYWKVRIWDEENRLVDYSKAQKFTIGESDNYIISTENKFVTDKIKPSKFENRDGVYFIDFGKAAFATME FNYQAKTPHTLTIRVGEMIDENGNVNRTPPAKSNIRYQELKVEVKPGQTRYRIPIQTDERNTRPNKAIPLPKGFPPLLPFRYAEIEGAQSSINANDVEQL AYHTFWDEKASSFKSDNNILNQVWDLSKYSIKATTFNGLYVDGDRERIPYEADAYLNQLSHYTTDREYAMARRTIEYFMKNPTWPTEWQQHVALLLYADY MYTGNTELVERYYEALKHKSLYELSNEDGLITSTKVDAEFMKKLGFPEGYKKPLTDIVDWPGANFNGSKTPGERDGFVFQPYNTVINSFFYENMKIMAQF AKILGKTDEVLDFELRAAKAKKAVNEQMFDKKRGIYVDGIGTDHASLHANMMPLAFGLVPQEHVDTVVEFVKSRGMACSVYGAQFLLDGLYNVGEADYAL DLLASTSERSWYNMIRIGSTITLEAWDNKYKNNLDWNHAWGAVPANAIPRGLWGIKPKTAGFGIASIKPQMGKLKSSQITVPTVRGAIHATFTHNGPRSQ TYEIEIPGNMVAEFSLDDIDGKDLIHNGQKVPAAFGAVQLSPGKHIIELKINSFGAGWSHPQFEKENLYFQSNLAYGSQGDLNPLINEIS
KIISAAGSFDVKEERTAASLLQLSGNASDFSYGRNSITLTTSA—

•  RED : Linker
•  GREEN : His Tag sequence
•  BLUE : TEV cleavage sequence
•  YELLOW : alpha-haemolysin signal sequence (hylA)