                         SEQUENCE LISTING

<110>  VIB VZW
       UNIVERSITEIT GENT
 
<120>  Improved aminopeptidases for single molecule peptide sequencing

<130>  NiCa/Npept/673

<150>  GB 1918108.0
<151>  2019-12-10

<160>  11    

<170>  PatentIn version 3.5

<210>  1
<211>  285
<212>  PRT
<213>  Streptomyces griseus

<400>  1

Met Ala Pro Asp Ile Pro Leu Ala Asn Val Lys Ala His Leu Thr Gln 
1               5                   10                  15      


Leu Ser Thr Ile Ala Ala Asn Asn Gly Gly Asn Arg Ala His Gly Arg 
            20                  25                  30          


Pro Gly Tyr Lys Ala Ser Val Asp Tyr Val Lys Ala Lys Leu Asp Ala 
        35                  40                  45              


Ala Gly Tyr Thr Thr Thr Leu Gln Gln Phe Thr Ser Gly Gly Ala Thr 
    50                  55                  60                  


Gly Tyr Asn Leu Ile Ala Asp Trp Pro Gly Gly Asp Pro Asn Lys Val 
65                  70                  75                  80  


Leu Met Ala Gly Ala His Leu Asp Ser Val Ser Ser Gly Ala Gly Ile 
                85                  90                  95      


Asn Asp Asn Gly Ser Gly Ser Ala Ala Val Leu Glu Thr Ala Leu Ala 
            100                 105                 110         


Val Ser Arg Ala Gly Tyr Gln Pro Asp Lys His Leu Arg Phe Ala Trp 
        115                 120                 125             


Trp Gly Ala Glu Glu Leu Gly Leu Ile Gly Ser Lys Tyr Tyr Val Asn 
    130                 135                 140                 


Asn Leu Pro Ser Ala Asp Arg Ser Lys Leu Ala Gly Tyr Leu Asn Phe 
145                 150                 155                 160 


Asp Met Ile Gly Ser Pro Asn Pro Gly Tyr Phe Val Tyr Asp Asp Asp 
                165                 170                 175     


Pro Val Ile Glu Lys Thr Phe Lys Asp Tyr Phe Ala Gly Leu Asn Val 
            180                 185                 190         


Pro Thr Glu Ile Glu Thr Glu Gly Asp Gly Arg Ser Asp His Ala Pro 
        195                 200                 205             


Phe Lys Asn Val Gly Val Pro Val Gly Gly Leu Phe Thr Gly Ala Gly 
    210                 215                 220                 


Tyr Thr Lys Ser Ala Ala Gln Ala Gln Lys Trp Gly Gly Thr Ala Gly 
225                 230                 235                 240 


Gln Ala Phe Asp Arg Cys Tyr His Ser Ser Cys Asp Ser Leu Ser Asn 
                245                 250                 255     


Ile Asn Asp Thr Ala Leu Asp Arg Asn Ser Asp Ala Ala Ala His Ala 
            260                 265                 270         


Ile Trp Thr Leu Ser Ser Gly Thr Gly Glu Pro Pro Thr 
        275                 280                 285 


<210>  2
<211>  299
<212>  PRT
<213>  Aeromonas proteolytica

<400>  2

Met Pro Pro Ile Thr Gln Gln Ala Thr Val Thr Ala Trp Leu Pro Gln 
1               5                   10                  15      


Val Asp Ala Ser Gln Ile Thr Gly Thr Ile Ser Ser Leu Glu Ser Phe 
            20                  25                  30          


Thr Asn Arg Phe Tyr Thr Thr Thr Ser Gly Ala Gln Ala Ser Asp Trp 
        35                  40                  45              


Ile Ala Ser Glu Trp Gln Ala Leu Ser Ala Ser Leu Pro Asn Ala Ser 
    50                  55                  60                  


Val Lys Gln Val Ser His Ser Gly Tyr Asn Gln Lys Ser Val Val Met 
65                  70                  75                  80  


Thr Ile Thr Gly Ser Glu Ala Pro Asp Glu Trp Ile Val Ile Gly Gly 
                85                  90                  95      


His Leu Asp Ser Thr Ile Gly Ser His Thr Asn Glu Gln Ser Val Ala 
            100                 105                 110         


Pro Gly Ala Asp Asp Asp Ala Ser Gly Ile Ala Ala Val Thr Glu Val 
        115                 120                 125             


Ile Arg Val Leu Ser Glu Asn Asn Phe Gln Pro Lys Arg Ser Ile Ala 
    130                 135                 140                 


Phe Met Ala Tyr Ala Ala Glu Glu Val Gly Leu Arg Gly Ser Gln Asp 
145                 150                 155                 160 


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


Gln Leu Asp Met Thr Asn Tyr Lys Gly Ser Ala Gln Asp Val Val Phe 
            180                 185                 190         


Ile Thr Asp Tyr Thr Asp Ser Asn Phe Thr Gln Tyr Leu Thr Gln Leu 
        195                 200                 205             


Met Asp Glu Tyr Leu Pro Ser Leu Thr Tyr Gly Phe Asp Thr Cys Gly 
    210                 215                 220                 


Tyr Ala Cys Ser Asp His Ala Ser Trp His Asn Ala Gly Tyr Pro Ala 
225                 230                 235                 240 


Ala Met Pro Phe Glu Ser Lys Phe Asn Asp Tyr Asn Pro Arg Ile His 
                245                 250                 255     


Thr Thr Gln Asp Thr Leu Ala Asn Ser Asp Pro Thr Gly Ser His Ala 
            260                 265                 270         


Lys Lys Phe Thr Gln Leu Gly Leu Ala Tyr Ala Ile Glu Met Gly Ser 
        275                 280                 285             


Ala Thr Gly Asp Thr Pro Thr Pro Gly Asn Gln 
    290                 295                 


<210>  3
<211>  317
<212>  PRT
<213>  Serratia marcescens

<400>  3

Met Glu Gln Leu Arg Gly Leu Tyr Pro Pro Leu Ala Ala Tyr Asp Ser 
1               5                   10                  15      


Gly Trp Leu Asp Thr Gly Asp Gly His Arg Ile Tyr Trp Glu Leu Ser 
            20                  25                  30          


Gly Asn Pro Asn Gly Lys Pro Ala Val Phe Ile His Gly Gly Pro Gly 
        35                  40                  45              


Gly Gly Ile Ser Pro His His Arg Gln Leu Phe Asp Pro Glu Arg Tyr 
    50                  55                  60                  


Lys Val Leu Leu Phe Asp Gln Arg Gly Cys Gly Arg Ser Arg Pro His 
65                  70                  75                  80  


Ala Ser Leu Asp Asn Asn Thr Thr Trp His Leu Val Ala Asp Ile Glu 
                85                  90                  95      


Arg Leu Arg Glu Met Ala Gly Val Glu Gln Trp Leu Val Phe Gly Gly 
            100                 105                 110         


Ser Trp Gly Ser Thr Leu Ala Leu Ala Tyr Ala Gln Thr His Pro Glu 
        115                 120                 125             


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


Arg Leu His Trp Tyr Tyr Gln Asp Gly Ala Ser Arg Phe Phe Pro Glu 
145                 150                 155                 160 


Lys Trp Glu Arg Val Leu Ser Ile Leu Ser Asp Asp Glu Arg Lys Asp 
                165                 170                 175     


Val Ile Ala Ala Tyr Arg Gln Arg Leu Thr Ser Ala Asp Pro Gln Val 
            180                 185                 190         


Gln Leu Glu Ala Ala Lys Leu Trp Ser Val Trp Glu Gly Glu Thr Val 
        195                 200                 205             


Thr Leu Leu Pro Ser Arg Glu Ser Ala Ser Phe Gly Glu Asp Asp Phe 
    210                 215                 220                 


Ala Leu Ala Phe Ala Arg Ile Glu Asn His Tyr Phe Thr His Leu Gly 
225                 230                 235                 240 


Phe Leu Glu Ser Asp Asp Gln Leu Leu Arg Asn Val Pro Leu Ile Arg 
                245                 250                 255     


His Ile Pro Ala Val Ile Val His Gly Arg Tyr Asp Met Ala Cys Gln 
            260                 265                 270         


Val Gln Asn Ala Trp Asp Leu Ala Lys Ala Trp Pro Glu Ala Glu Leu 
        275                 280                 285             


His Ile Val Glu Gly Ala Gly His Ser Tyr Asp Glu Pro Gly Ile Leu 
    290                 295                 300                 


His Gln Leu Met Ile Ala Thr Asp Arg Phe Ala Gly Lys 
305                 310                 315         


<210>  4
<211>  295
<212>  PRT
<213>  Pyrococcus furiosus

<400>  4

Met Asp Thr Glu Lys Leu Met Lys Ala Gly Glu Ile Ala Lys Lys Val 
1               5                   10                  15      


Arg Glu Lys Ala Ile Lys Leu Ala Arg Pro Gly Met Leu Leu Leu Glu 
            20                  25                  30          


Leu Ala Glu Ser Ile Glu Lys Met Ile Met Glu Leu Gly Gly Lys Pro 
        35                  40                  45              


Ala Phe Pro Val Asn Leu Ser Ile Asn Glu Ile Ala Ala His Tyr Thr 
    50                  55                  60                  


Pro Tyr Lys Gly Asp Thr Thr Val Leu Lys Glu Gly Asp Tyr Leu Lys 
65                  70                  75                  80  


Ile Asp Val Gly Val His Ile Asp Gly Phe Ile Ala Asp Thr Ala Val 
                85                  90                  95      


Thr Val Arg Val Gly Met Glu Glu Asp Glu Leu Met Glu Ala Ala Lys 
            100                 105                 110         


Glu Ala Leu Asn Ala Ala Ile Ser Val Ala Arg Ala Gly Val Glu Ile 
        115                 120                 125             


Lys Glu Leu Gly Lys Ala Ile Glu Asn Glu Ile Arg Lys Arg Gly Phe 
    130                 135                 140                 


Lys Pro Ile Val Asn Leu Ser Gly His Lys Ile Glu Arg Tyr Lys Leu 
145                 150                 155                 160 


His Ala Gly Ile Ser Ile Pro Asn Ile Tyr Arg Pro His Asp Asn Tyr 
                165                 170                 175     


Val Leu Lys Glu Gly Asp Val Phe Ala Ile Glu Pro Phe Ala Thr Ile 
            180                 185                 190         


Gly Ala Gly Gln Val Ile Glu Val Pro Pro Thr Leu Ile Tyr Met Tyr 
        195                 200                 205             


Val Arg Asp Val Pro Val Arg Val Ala Gln Ala Arg Phe Leu Leu Ala 
    210                 215                 220                 


Lys Ile Lys Arg Glu Tyr Gly Thr Leu Pro Phe Ala Tyr Arg Trp Leu 
225                 230                 235                 240 


Gln Asn Asp Met Pro Glu Gly Gln Leu Lys Leu Ala Leu Lys Thr Leu 
                245                 250                 255     


Glu Lys Ala Gly Ala Ile Tyr Gly Tyr Pro Val Leu Lys Glu Ile Arg 
            260                 265                 270         


Asn Gly Ile Val Ala Gln Phe Glu His Thr Ile Ile Val Glu Lys Asp 
        275                 280                 285             


Ser Val Ile Val Thr Thr Glu 
    290                 295 


<210>  5
<211>  793
<212>  PRT
<213>  Lactobacillus helveticus

<400>  5

Met Lys Tyr Asn Gln Tyr Ala Tyr Val Glu Thr Asp Phe Gln Gln Gln 
1               5                   10                  15      


Val Lys Glu Leu Ile Asp Ile Asn Phe Leu Pro Lys Asn Tyr Gln Val 
            20                  25                  30          


Trp Asp Phe Ser Ser Leu Leu Ala Lys Leu Val Lys Asn Ala Ile Ala 
        35                  40                  45              


Glu Ala Lys Thr Asp Ala Ala Lys Asn Ala Lys Leu Ala Glu Phe Ala 
    50                  55                  60                  


Val Ser Asp His Gln Thr Leu Ala Asp Phe Leu Lys Glu Lys Pro Thr 
65                  70                  75                  80  


Glu Ile Gly Thr Lys Gln Phe Tyr Asn Val Ala Leu Gln Leu Leu Gly 
                85                  90                  95      


Tyr His Val His Tyr Asp Tyr Asp Phe Ala Asp Pro Thr Gly Phe Met 
            100                 105                 110         


Gln Arg Asn Ala Leu Pro Phe Leu Gln Asp Ile Ser Asp Asn Gln Lys 
        115                 120                 125             


Leu Ile Ser Ala Phe Tyr Arg Leu Leu Asn Thr Arg Ala Lys Asn Gly 
    130                 135                 140                 


Gln Ile Leu Leu Asp Val Met Ala Gly Lys Gly Tyr Phe Thr Gln Phe 
145                 150                 155                 160 


Trp Gly Gln Asn Lys Phe Lys Phe Phe Asn Gly Lys Ser Ile Pro Val 
                165                 170                 175     


Phe Asp Thr Asn Lys Val Ile Arg Glu Val Val Tyr Val Glu Thr Asp 
            180                 185                 190         


Leu Asp Thr Asp His Asp Gly Lys Ser Asp Leu Ile Gln Val Thr Val 
        195                 200                 205             


Phe Arg Pro Glu Glu Thr Asn Lys Gly Leu Lys Val Pro Ala Leu Tyr 
    210                 215                 220                 


Thr Ala Ser Pro Tyr Phe Gly Gly Ile Ile Ala Asn Glu Lys Arg Asn 
225                 230                 235                 240 


His Asn Val Asp Glu Asn Leu Ser Asp Ser Thr Glu Trp Asn Asp Pro 
                245                 250                 255     


Gln Tyr Val His Ser Pro Ile Val Lys Ala Glu Lys Pro Asp Gly Ser 
            260                 265                 270         


Ser Arg Pro Ala Thr Glu Glu Ala Val His Lys Ser Ser Tyr Pro Leu 
        275                 280                 285             


Asn Glu Tyr Met Leu Ala Arg Gly Phe Ala Ser Val Phe Ala Gly Ala 
    290                 295                 300                 


Ile Gly Thr Arg Gly Ser Asp Gly Val Arg Ile Thr Gly Ala Pro Glu 
305                 310                 315                 320 


Glu Thr Glu Ser Ala Ala Ala Val Ile Glu Trp Leu His Gly Asp Arg 
                325                 330                 335     


Val Ala Tyr Thr Asp Arg Thr Arg Thr Val Gln Thr Thr Ala Asp Trp 
            340                 345                 350         


Cys Asn Gly Asn Ile Gly Met Thr Gly Arg Ser Tyr Leu Gly Thr Leu 
        355                 360                 365             


Gln Ile Ala Ile Ala Thr Thr Gly Val Lys Gly Leu Lys Thr Val Val 
    370                 375                 380                 


Ser Glu Ala Ala Ile Ser Ser Trp Tyr Asp Tyr Tyr Arg Glu His Gly 
385                 390                 395                 400 


Leu Val Ile Ala Pro Glu Ala Cys Gln Gly Glu Asp Leu Asp Leu Leu 
                405                 410                 415     


Ala Glu Thr Cys Gln Ser Asn Leu Trp Asp Ala Gly Ser Tyr Leu Lys 
            420                 425                 430         


Ile Lys Pro Glu Tyr Asp Lys Met Gln Lys Gln Leu Arg Glu Lys Glu 
        435                 440                 445             


Asp Arg Asn Thr Gly Gln Tyr Ser Asp Phe Trp Glu Ala Arg Asn Tyr 
    450                 455                 460                 


Arg His His Ala Asp Gly Ile Lys Cys Ser Trp Ile Ser Val His Gly 
465                 470                 475                 480 


Leu Asn Asp Trp Asn Val Lys Pro Lys Asn Val Tyr Lys Ile Trp Gln 
                485                 490                 495     


Leu Val Lys Lys Met Pro Met Lys His His Leu Phe Leu His Gln Gly 
            500                 505                 510         


Pro His Tyr Asn Met Asn Asn Leu Val Ser Ile Asp Phe Thr Asp Leu 
        515                 520                 525             


Met Asn Leu Trp Phe Val His Glu Leu Leu Gly Ile Glu Asn Asn Ala 
    530                 535                 540                 


Tyr Asn Gln Trp Pro Thr Val Met Ile Gln Asp Asn Leu Gln Ala Asp 
545                 550                 555                 560 


Lys Trp His Glu Glu Pro Asp Trp Ser Asn Asp Leu Gly Gln Glu Lys 
                565                 570                 575     


Ile Tyr Tyr Pro Thr Asp Glu Gly Glu Leu Phe Gln Asp Gly Asn Gly 
            580                 585                 590         


Lys Ala Gln Lys Ser Phe Thr Asp Val Gly Gly Ile Glu Phe Lys Lys 
        595                 600                 605             


Ala Gly Ile Ser Glu Ser Asp Trp Gln Tyr Lys Phe Ile Cys Gly Asp 
    610                 615                 620                 


Glu Lys Trp Ala Lys Pro Ser Leu Arg Phe Glu Thr Asp Glu Phe Thr 
625                 630                 635                 640 


His Pro Thr Thr Ile Val Gly Arg Pro Glu Val Lys Val Arg Val Ser 
                645                 650                 655     


Ala Ser Leu Pro Lys Gly Glu Ile Ser Val Ala Leu Val Glu Leu Gly 
            660                 665                 670         


Glu Arg Gln Arg Leu Thr Ala Thr Pro Lys Phe Leu Met His Gly Gly 
        675                 680                 685             


Gln Glu Leu Gly Tyr Arg Phe Gly Thr Asp Thr Leu Gln Glu Phe Val 
    690                 695                 700                 


Pro Asp Lys Lys Thr Lys Ala Lys Leu Ile Thr Lys Ala His Met Asn 
705                 710                 715                 720 


Leu Gln Asn Phe Lys Asp Met Lys Lys Pro Glu Ala Ile Asp Ala Asp 
                725                 730                 735     


Lys Phe Tyr Asp Leu Asp Phe Leu Leu Gln Pro Thr Tyr Tyr Thr Ile 
            740                 745                 750         


Pro Ser Gly Ser Lys Leu Ala Leu Ile Ile Tyr Ser Thr Asp Gln Gly 
        755                 760                 765             


Met Thr Lys Arg Pro Leu Glu Asp Glu Thr Tyr Thr Ile Asp Leu Ala 
    770                 775                 780                 


Asn Thr Glu Ile Lys Phe Tyr Glu Lys 
785                 790             


<210>  6
<211>  569
<212>  PRT
<213>  Streptomyces griseus

<400>  6

Met Gly Arg Thr Thr Leu Val Glu His Asp Val Pro Val Glu Met Arg 
1               5                   10                  15      


Asp Gly Thr Val Leu Arg Ala Asp Val Trp Arg Pro Ala Asp Gly Pro 
            20                  25                  30          


Pro Ser Pro Ala Val Leu Phe Arg Thr Pro Tyr Gly Lys Ser Pro Leu 
        35                  40                  45              


Gly Leu Ala Thr Leu Thr Pro Ala Gln Cys Val Asp Arg Gly Tyr Ala 
    50                  55                  60                  


Ala Val Val Gln Asp Thr Arg Gly Arg Phe Gly Ser Glu Gly Glu Trp 
65                  70                  75                  80  


Ala Pro Leu Asp Trp Ser Arg Glu Gly Pro Asp Gly Tyr Asp Thr Val 
                85                  90                  95      


Glu Trp Thr Ala Gly Gln Pro Trp Cys Asp Gly Asn Val Ala Met Ala 
            100                 105                 110         


Gly Thr Ser Tyr Gln Ala Ile Val Gln Trp Leu Ala Ala Met Glu Lys 
        115                 120                 125             


Pro Pro His Leu Arg Ala Ile Ala Pro Thr Met Ser Thr Ser Ala Pro 
    130                 135                 140                 


Phe Asp Ala Glu Gln Leu Gly Gly Ser Leu Arg Leu Asp His Leu Thr 
145                 150                 155                 160 


Ser Trp Leu Gly Leu Thr Ala Leu Glu Trp Val Gln Arg Arg Ala Ala 
                165                 170                 175     


Ala Gly Asp Pro Val Asp Gly Ala Val Val Ala Glu Val Val Gln Leu 
            180                 185                 190         


Leu Thr Thr Pro Asp Val Pro Leu Arg Arg Trp Pro Leu Ser Thr Ile 
        195                 200                 205             


Leu Asp Phe Glu Gly Phe Pro Gly Arg Leu Arg Asp Ile Phe Ala Gly 
    210                 215                 220                 


Lys Val Ala Thr Val Ala Asp Tyr Arg Leu Gly Glu Val Gly Val Pro 
225                 230                 235                 240 


Thr Phe Ser Val Gly Gly Trp Tyr Asp Val Phe Ser His Gly Thr Ile 
                245                 250                 255     


Glu Leu His Arg Ala Met Arg Ala Gln Asp Pro Val Ala Gly Arg His 
            260                 265                 270         


Glu Leu Val Val Gly Pro Trp Val His Ser Gly Gln Leu Pro Gln Val 
        275                 280                 285             


Gln Gly Glu Val Asn Thr Gly Pro Tyr Gly Ser Ala Gln Gly Ala Arg 
    290                 295                 300                 


Leu Ala Asp Leu His Leu Asp Phe Phe Asp Arg His Leu Arg Pro Ser 
305                 310                 315                 320 


Ala Ala Asp Ala Gly Thr Ala Ala Gly Gly Asp Val Arg Tyr Phe Leu 
                325                 330                 335     


Phe Gly Asp Asp Thr Trp His Arg Ala Glu Ser Trp Pro Pro Pro Glu 
            340                 345                 350         


Ala Val Asp Ser Pro Trp Tyr Leu Ala Gly Pro Glu Gly Gly Glu Glu 
        355                 360                 365             


Asp Gly Arg Leu Leu Pro Glu Pro Pro Gln Glu Gly Phe Gly Gln Asp 
    370                 375                 380                 


Ala Phe Thr Tyr Asp Pro Glu Asn Pro Val Pro Ser His Gly Gly Arg 
385                 390                 395                 400 


Val Leu Gln Leu Gly Arg Leu Ala Ala Gly Pro Leu Asn Gln Ala His 
                405                 410                 415     


Leu Glu Ala Arg Pro Asp Val Leu Leu Tyr Thr Ser Glu Pro Leu Thr 
            420                 425                 430         


Glu Pro Leu Asp Val Val Gly Arg Gln Arg Leu Arg Leu Arg Phe Gly 
        435                 440                 445             


Ser Asp Ala Pro Ala Thr Ala Val Val Ala Lys Leu Thr Asp Val His 
    450                 455                 460                 


Pro Asp Gly Arg Ser Leu Leu Val Ala Glu Ala Ser Leu Arg Leu Asp 
465                 470                 475                 480 


Thr Gly Asn Gly Val Pro Pro Asp Glu Pro Tyr Asp Ala Asp Leu Leu 
                485                 490                 495     


Leu Gly Asp Thr Ala Trp Arg Phe Ala Pro Gly His Arg Leu Arg Val 
            500                 505                 510         


His Val Thr Ser Ser Asn Phe Pro His Leu Asp Arg His Pro Asn Val 
        515                 520                 525             


Pro Gly Pro Val Gly Glu Ala Glu His Cys Arg Pro Ala Arg Gln Thr 
    530                 535                 540                 


Val Trp Tyr Gly Gly Pro Tyr Glu Ser Val Leu Arg Leu Arg Val Leu 
545                 550                 555                 560 


Pro Ala Pro Ser Thr Gly Gly Ile Ala 
                565                 


<210>  7
<211>  10
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  synthetic peptide

<400>  7

Ala Ala Ala Gly Gly Asn Asn Gly Gly Cys 
1               5                   10  


<210>  8
<211>  7
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  synthetic peptide

<400>  8

Gly Gly Asn Asn Gly Gly Lys 
1               5           


<210>  9
<211>  6
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  synthetic peptide

<400>  9

Leu Ala Gly Gly Asn Lys 
1               5       


<210>  10
<211>  5
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  synthetic peptide

<400>  10

Ala Gly Gly Asn Lys 
1               5   


<210>  11
<211>  4
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  synthetic peptide

<400>  11

Gly Gly Asn Lys 
1               


