                         SEQUENCE LISTING

<110>  INSTITUTO DE MEDICINA MOLECULAR JOAO LOBO ANTUNES
       BERNARDES, Goncalo Jose Lopes
       DE ALBUQUERQUE, Maria Ines Sousa
 
<120>  Production of Cross-reactive material 197 Fusion Proteins

<130>  008282519

<140>  PCT/EP2022/066394
<141>  2022-06-15

<150>  GB2108650.9
<151>  2021-06-17

<160>  6     

<170>  PatentIn version 3.5

<210>  1
<211>  535
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Full-length native CRM197 polypeptide

<400>  1

Gly Ala Asp Asp Val Val Asp Ser Ser Lys Ser Phe Val Met Glu Asn 
1               5                   10                  15      


Phe Ser Ser Tyr His Gly Thr Lys Pro Gly Tyr Val Asp Ser Ile Gln 
            20                  25                  30          


Lys Gly Ile Gln Lys Pro Lys Ser Gly Thr Gln Gly Asn Tyr Asp Asp 
        35                  40                  45              


Asp Trp Lys Glu Phe Tyr Ser Thr Asp Asn Lys Tyr Asp Ala Ala Gly 
    50                  55                  60                  


Tyr Ser Val Asp Asn Glu Asn Pro Leu Ser Gly Lys Ala Gly Gly Val 
65                  70                  75                  80  


Val Lys Val Thr Tyr Pro Gly Leu Thr Lys Val Leu Ala Leu Lys Val 
                85                  90                  95      


Asp Asn Ala Glu Thr Ile Lys Lys Glu Leu Gly Leu Ser Leu Thr Glu 
            100                 105                 110         


Pro Leu Met Glu Gln Val Gly Thr Glu Glu Phe Ile Lys Arg Phe Gly 
        115                 120                 125             


Asp Gly Ala Ser Arg Val Val Leu Ser Leu Pro Phe Ala Glu Gly Ser 
    130                 135                 140                 


Ser Ser Val Glu Tyr Ile Asn Asn Trp Glu Gln Ala Lys Ala Leu Ser 
145                 150                 155                 160 


Val Glu Leu Glu Ile Asn Phe Glu Thr Arg Gly Lys Arg Gly Gln Asp 
                165                 170                 175     


Ala Met Tyr Glu Tyr Met Ala Gln Ala Cys Ala Gly Asn Arg Val Arg 
            180                 185                 190         


Arg Ser Val Gly Ser Ser Leu Ser Cys Ile Asn Leu Asp Trp Asp Val 
        195                 200                 205             


Ile Arg Asp Lys Thr Lys Thr Lys Ile Glu Ser Leu Lys Glu His Gly 
    210                 215                 220                 


Pro Ile Lys Asn Lys Met Ser Glu Ser Pro Asn Lys Thr Val Ser Glu 
225                 230                 235                 240 


Glu Lys Ala Lys Gln Tyr Leu Glu Glu Phe His Gln Thr Ala Leu Glu 
                245                 250                 255     


His Pro Glu Leu Ser Glu Leu Lys Thr Val Thr Gly Thr Asn Pro Val 
            260                 265                 270         


Phe Ala Gly Ala Asn Tyr Ala Ala Trp Ala Val Asn Val Ala Gln Val 
        275                 280                 285             


Ile Asp Ser Glu Thr Ala Asp Asn Leu Glu Lys Thr Thr Ala Ala Leu 
    290                 295                 300                 


Ser Ile Leu Pro Gly Ile Gly Ser Val Met Gly Ile Ala Asp Gly Ala 
305                 310                 315                 320 


Val His His Asn Thr Glu Glu Ile Val Ala Gln Ser Ile Ala Leu Ser 
                325                 330                 335     


Ser Leu Met Val Ala Gln Ala Ile Pro Leu Val Gly Glu Leu Val Asp 
            340                 345                 350         


Ile Gly Phe Ala Ala Tyr Asn Phe Val Glu Ser Ile Ile Asn Leu Phe 
        355                 360                 365             


Gln Val Val His Asn Ser Tyr Asn Arg Pro Ala Tyr Ser Pro Gly His 
    370                 375                 380                 


Lys Thr Gln Pro Phe Leu His Asp Gly Tyr Ala Val Ser Trp Asn Thr 
385                 390                 395                 400 


Val Glu Asp Ser Ile Ile Arg Thr Gly Phe Gln Gly Glu Ser Gly His 
                405                 410                 415     


Asp Ile Lys Ile Thr Ala Glu Asn Thr Pro Leu Pro Ile Ala Gly Val 
            420                 425                 430         


Leu Leu Pro Thr Ile Pro Gly Lys Leu Asp Val Asn Lys Ser Lys Thr 
        435                 440                 445             


His Ile Ser Val Asn Gly Arg Lys Ile Arg Met Arg Cys Arg Ala Ile 
    450                 455                 460                 


Asp Gly Asp Val Thr Phe Cys Arg Pro Lys Ser Pro Val Tyr Val Gly 
465                 470                 475                 480 


Asn Gly Val His Ala Asn Leu His Val Ala Phe His Arg Ser Ser Ser 
                485                 490                 495     


Glu Lys Ile His Ser Asn Glu Ile Ser Ser Asp Ser Ile Gly Val Leu 
            500                 505                 510         


Gly Tyr Gln Lys Thr Val Asp His Thr Lys Val Asn Ser Lys Leu Ser 
        515                 520                 525             


Leu Phe Phe Glu Ile Lys Ser 
    530                 535 


<210>  2
<211>  5
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Enteropeptidase recognition site

<400>  2

Asp Asp Asp Asp Lys 
1               5   


<210>  3
<211>  553
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Complete iCRM197 fusion protein

<400>  3

Met Ala Ser Trp Ser His Pro Gln Phe Glu Lys Gly Ala Asp Asp Asp 
1               5                   10                  15      


Asp Lys Gly Ala Asp Asp Val Val Asp Ser Ser Lys Ser Phe Val Met 
            20                  25                  30          


Glu Asn Phe Ser Ser Tyr His Gly Thr Lys Pro Gly Tyr Val Asp Ser 
        35                  40                  45              


Ile Gln Lys Gly Ile Gln Lys Pro Lys Ser Gly Thr Gln Gly Asn Tyr 
    50                  55                  60                  


Asp Asp Asp Trp Lys Glu Phe Tyr Ser Thr Asp Asn Lys Tyr Asp Ala 
65                  70                  75                  80  


Ala Gly Tyr Ser Val Asp Asn Glu Asn Pro Leu Ser Gly Lys Ala Gly 
                85                  90                  95      


Gly Val Val Lys Val Thr Tyr Pro Gly Leu Thr Lys Val Leu Ala Leu 
            100                 105                 110         


Lys Val Asp Asn Ala Glu Thr Ile Lys Lys Glu Leu Gly Leu Ser Leu 
        115                 120                 125             


Thr Glu Pro Leu Met Glu Gln Val Gly Thr Glu Glu Phe Ile Lys Arg 
    130                 135                 140                 


Phe Gly Asp Gly Ala Ser Arg Val Val Leu Ser Leu Pro Phe Ala Glu 
145                 150                 155                 160 


Gly Ser Ser Ser Val Glu Tyr Ile Asn Asn Trp Glu Gln Ala Lys Ala 
                165                 170                 175     


Leu Ser Val Glu Leu Glu Ile Asn Phe Glu Thr Arg Gly Lys Arg Gly 
            180                 185                 190         


Gln Asp Ala Met Tyr Glu Tyr Met Ala Gln Ala Cys Ala Gly Asn Arg 
        195                 200                 205             


Val Arg Arg Ser Val Gly Ser Ser Leu Ser Cys Ile Asn Leu Asp Trp 
    210                 215                 220                 


Asp Val Ile Arg Asp Lys Thr Lys Thr Lys Ile Glu Ser Leu Lys Glu 
225                 230                 235                 240 


His Gly Pro Ile Lys Asn Lys Met Ser Glu Ser Pro Asn Lys Thr Val 
                245                 250                 255     


Ser Glu Glu Lys Ala Lys Gln Tyr Leu Glu Glu Phe His Gln Thr Ala 
            260                 265                 270         


Leu Glu His Pro Glu Leu Ser Glu Leu Lys Thr Val Thr Gly Thr Asn 
        275                 280                 285             


Pro Val Phe Ala Gly Ala Asn Tyr Ala Ala Trp Ala Val Asn Val Ala 
    290                 295                 300                 


Gln Val Ile Asp Ser Glu Thr Ala Asp Asn Leu Glu Lys Thr Thr Ala 
305                 310                 315                 320 


Ala Leu Ser Ile Leu Pro Gly Ile Gly Ser Val Met Gly Ile Ala Asp 
                325                 330                 335     


Gly Ala Val His His Asn Thr Glu Glu Ile Val Ala Gln Ser Ile Ala 
            340                 345                 350         


Leu Ser Ser Leu Met Val Ala Gln Ala Ile Pro Leu Val Gly Glu Leu 
        355                 360                 365             


Val Asp Ile Gly Phe Ala Ala Tyr Asn Phe Val Glu Ser Ile Ile Asn 
    370                 375                 380                 


Leu Phe Gln Val Val His Asn Ser Tyr Asn Arg Pro Ala Tyr Ser Pro 
385                 390                 395                 400 


Gly His Lys Thr Gln Pro Phe Leu His Asp Gly Tyr Ala Val Ser Trp 
                405                 410                 415     


Asn Thr Val Glu Asp Ser Ile Ile Arg Thr Gly Phe Gln Gly Glu Ser 
            420                 425                 430         


Gly His Asp Ile Lys Ile Thr Ala Glu Asn Thr Pro Leu Pro Ile Ala 
        435                 440                 445             


Gly Val Leu Leu Pro Thr Ile Pro Gly Lys Leu Asp Val Asn Lys Ser 
    450                 455                 460                 


Lys Thr His Ile Ser Val Asn Gly Arg Lys Ile Arg Met Arg Cys Arg 
465                 470                 475                 480 


Ala Ile Asp Gly Asp Val Thr Phe Cys Arg Pro Lys Ser Pro Val Tyr 
                485                 490                 495     


Val Gly Asn Gly Val His Ala Asn Leu His Val Ala Phe His Arg Ser 
            500                 505                 510         


Ser Ser Glu Lys Ile His Ser Asn Glu Ile Ser Ser Asp Ser Ile Gly 
        515                 520                 525             


Val Leu Gly Tyr Gln Lys Thr Val Asp His Thr Lys Val Asn Ser Lys 
    530                 535                 540                 


Leu Ser Leu Phe Phe Glu Ile Lys Ser 
545                 550             


<210>  4
<211>  1659
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Complete iCRM197 fusion protein

<400>  4
atggcaagct ggagccaccc gcagttcgaa aagggtgcag atgacgacga caagggtgca       60

gatgatgttg ttgatagcag caaaagcttt gtgatggaaa actttagcag ctaccatggc      120

accaaaccgg gttatgttga tagcattcag aaaggtattc agaaaccgaa aagcggcacc      180

cagggtaatt atgatgatga ttggaaagag ttctacagca ccgataacaa atatgatgca      240

gcaggttata gcgtggataa tgaaaatccg ctgagcggta aagccggtgg tgttgttaaa      300

gttacctatc cgggtctgac caaagttctg gcactgaaag ttgataatgc cgaaaccatc      360

aaaaaagaac tgggtctgag cctgaccgaa ccgctgatgg aacaggttgg caccgaagaa      420

tttatcaaac gttttggtga tggtgcaagc cgtgttgttc tgagcctgcc gtttgcagaa      480

ggtagcagca gcgttgaata tatcaataat tgggaacagg caaaagccct gagcgttgaa      540

ctggaaatca attttgaaac ccgtggtaaa cgtggtcagg atgcaatgta tgaatacatg      600

gcacaggcat gtgcaggtaa tcgtgttcgt cgtagcgttg gtagcagcct gagctgtatt      660

aatctggatt gggatgtgat tcgcgacaaa accaaaacca aaatcgaaag cctgaaagaa      720

catggtccga ttaaaaacaa aatgagcgaa agcccgaata aaaccgtgag cgaagaaaaa      780

gcaaaacagt atctggaaga atttcatcag accgcactgg aacatccgga actgagcgaa      840

ctgaaaaccg ttaccggcac caatccggtt tttgccggtg caaattatgc agcatgggca      900

gttaatgttg cacaggttat tgatagcgaa accgcagata atctggaaaa aaccaccgca      960

gcactgagca ttctgcctgg tattggtagc gttatgggta ttgcagatgg tgcagttcat     1020

cataacaccg aagaaattgt tgcacagagc attgcactga gcagcctgat ggttgcacag     1080

gcaattccgc tggttggtga actggttgat attggttttg cagcctataa ctttgtcgag     1140

agcattatca acctgtttca ggttgtgcat aacagctata atcgtccggc atatagtccg     1200

ggtcataaaa cccagccgtt tctgcatgat ggttatgcag ttagctggaa taccgttgaa     1260

gatagcatta ttcgtaccgg ttttcagggt gaaagcggtc atgatatcaa aattaccgca     1320

gaaaatacac cgctgccgat tgccggtgtt ctgctgccga ccattccggg taaactggat     1380

gtgaataaaa gcaaaaccca tatcagcgtg aacggtcgta aaattcgtat gcgttgtcgt     1440

gcaattgatg gtgatgttac cttttgtcgt ccgaaaagtc cggtttatgt tggtaatggt     1500

gttcatgcaa atctgcatgt tgcatttcat cgtagctcca gcgaaaaaat tcatagcaat     1560

gaaattagca gcgatagcat tggtgttctg ggttatcaga aaaccgttga tcataccaaa     1620

gtgaacagca aactgagcct gttttttgaa atcaaaagc                            1659


<210>  5
<211>  8
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Tag

<400>  5

Trp Ser His Pro Gln Phe Glu Lys 
1               5               


<210>  6
<211>  7
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Linker

<400>  6

Gly Ala Asp Asp Asp Asp Lys 
1               5           


