                         SEQUENCE LISTING

<110>  Rijksuniversiteit Groningen
 
<120>  Artificial nanopores and uses and methods relating thereto

<130>  P125030PC00

<140>  PCT/NL2020/050726
<141>  2020-11-19

<150>  EP 19210168.1
<151>  2019-11-19

<160>  36    

<170>  PatentIn version 3.5

<210>  1
<211>  28
<212>  PRT
<213>  Actinia fragacea

<400>  1

Ser Ala Asp Val Ala Gly Ala Val Ile Asp Gly Ala Gly Leu Gly Phe 
1               5                   10                  15      


Asp Val Leu Lys Thr Val Leu Glu Ala Leu Gly Asn 
            20                  25              


<210>  2
<211>  44
<212>  PRT
<213>  Escherichia coli

<400>  2

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


Asp Lys Thr Val Glu Val Val Lys Asn Ala Ile Glu Thr Ala Asp Gly 
            20                  25                  30          


Ala Leu Asp Leu Tyr Asn Lys Tyr Leu Asp Gln Val 
        35                  40                  


<210>  3
<211>  18
<212>  PRT
<213>  Yersinia enterocolitica

<400>  3

Phe Thr Gly Ala Ile Gly Gly Ile Ile Ala Met Ala Ile Thr Gly Gly 
1               5                   10                  15      


Ile Phe 
        


<210>  4
<211>  42
<212>  PRT
<213>  Yersinia enterocolitica

<400>  4

Leu Val Asp Ala Phe Lys Asp Leu Ile Pro Thr Gly Glu Asn Leu Ser 
1               5                   10                  15      


Glu Leu Asp Leu Ala Lys Pro Glu Ile Glu Leu Leu Lys Gln Ser Leu 
            20                  25                  30          


Glu Ile Thr Lys Lys Leu Leu Gly Gln Phe 
        35                  40          


<210>  5
<211>  35
<212>  PRT
<213>  Escherichia coli

<400>  5

Ala Pro Leu Val Arg Trp Asn Arg Val Ile Ser Gln Leu Val Pro Thr 
1               5                   10                  15      


Ile Ser Gly Val His Asp Met Thr Glu Thr Val Arg Tyr Ile Lys Arg 
            20                  25                  30          


Trp Pro Asn 
        35  


<210>  6
<211>  22
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  TM sequence

<400>  6

Val His Gly Asn Ala Glu Val His Ala Ser Phe Phe Asp Ile Gly Gly 
1               5                   10                  15      


Ser Val Ser Ala Gly Phe 
            20          


<210>  7
<211>  28
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  TM sequence with linkers

<400>  7

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


Ile Gly Gly Ser Val Ser Ala Gly Phe Ser Ser Gly 
            20                  25              


<210>  8
<211>  37
<212>  PRT
<213>  Thermoplasma acidophilum

<400>  8

Pro Val Pro Asp Pro Val Lys Glu Lys Glu Lys Glu Glu Arg Lys Lys 
1               5                   10                  15      


Gln Gln Glu Lys Glu Glu Lys Glu Glu Lys Lys Lys Gly Asp Glu Asp 
            20                  25                  30          


Asp Lys Gly Pro Pro 
        35          


<210>  9
<211>  251
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  subunit of artificial PA28 nanopore

<400>  9

Met Ala Thr Leu Arg Val His Pro Glu Ala Gln Ala Lys Val Asp Val 
1               5                   10                  15      


Phe Arg Glu Asp Leu Cys Ser Lys Thr Glu Asn Leu Leu Gly Ser Tyr 
            20                  25                  30          


Phe Pro Lys Lys Ile Ser Glu Leu Asp Ala Phe Leu Lys Glu Pro Ala 
        35                  40                  45              


Leu Asn Glu Ala Asn Leu Ser Asn Leu Lys Ala Pro Leu Asp Ile Gly 
    50                  55                  60                  


Ser Ser Val His Gly Asn Ala Glu Val His Ala Ser Phe Phe Asp Ile 
65                  70                  75                  80  


Gly Gly Ser Val Ser Ala Gly Phe Ser Ser Gly Cys Gly Pro Val Asn 
                85                  90                  95      


Cys Asn Glu Lys Ile Val Val Leu Leu Gln Arg Leu Lys Pro Glu Ile 
            100                 105                 110         


Lys Asp Val Thr Glu Gln Leu Asn Leu Val Thr Thr Trp Leu Gln Leu 
        115                 120                 125             


Gln Ile Pro Arg Ile Glu Asp Gly Asn Asn Phe Gly Val Ala Val Gln 
    130                 135                 140                 


Glu Lys Val Phe Glu Leu Met Thr Asn Leu His Thr Lys Leu Glu Gly 
145                 150                 155                 160 


Phe His Thr Gln Ile Ser Lys Tyr Phe Ser Glu Arg Gly Asp Ala Val 
                165                 170                 175     


Ala Lys Ala Ala Lys Gln Pro His Val Gly Asp Tyr Arg Gln Leu Val 
            180                 185                 190         


His Glu Leu Asp Glu Ala Glu Tyr Gln Glu Ile Arg Leu Met Val Met 
        195                 200                 205             


Glu Ile Arg Asn Ala Tyr Ala Val Leu Tyr Asp Ile Ile Leu Lys Asn 
    210                 215                 220                 


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


Gly Ser Ser Trp Ser His Pro Gln Phe Glu Lys 
                245                 250     


<210>  10
<211>  260
<212>  PRT
<213>  Thermoplasma acidophilum

<400>  10

Met Ala Thr Leu Arg Val His Pro Glu Ala Gln Ala Lys Val Asp Val 
1               5                   10                  15      


Phe Arg Glu Asp Leu Cys Ser Lys Thr Glu Asn Leu Leu Gly Ser Tyr 
            20                  25                  30          


Phe Pro Lys Lys Ile Ser Glu Leu Asp Ala Phe Leu Lys Glu Pro Ala 
        35                  40                  45              


Leu Asn Glu Ala Asn Leu Ser Asn Leu Lys Ala Pro Leu Asp Ile Pro 
    50                  55                  60                  


Val Pro Asp Pro Val Lys Glu Lys Glu Lys Glu Glu Arg Lys Lys Gln 
65                  70                  75                  80  


Gln Glu Lys Glu Glu Lys Glu Glu Lys Lys Lys Gly Asp Glu Asp Asp 
                85                  90                  95      


Lys Gly Pro Pro Cys Gly Pro Val Asn Cys Asn Glu Lys Ile Val Val 
            100                 105                 110         


Leu Leu Gln Arg Leu Lys Pro Glu Ile Lys Asp Val Thr Glu Gln Leu 
        115                 120                 125             


Asn Leu Val Thr Thr Trp Leu Gln Leu Gln Ile Pro Arg Ile Glu Asp 
    130                 135                 140                 


Gly Asn Asn Phe Gly Val Ala Val Gln Glu Lys Val Phe Glu Leu Met 
145                 150                 155                 160 


Thr Asn Leu His Thr Lys Leu Glu Gly Phe His Thr Gln Ile Ser Lys 
                165                 170                 175     


Tyr Phe Ser Glu Arg Gly Asp Ala Val Ala Lys Ala Ala Lys Gln Pro 
            180                 185                 190         


His Val Gly Asp Tyr Arg Gln Leu Val His Glu Leu Asp Glu Ala Glu 
        195                 200                 205             


Tyr Gln Glu Ile Arg Leu Met Val Met Glu Ile Arg Asn Ala Tyr Ala 
    210                 215                 220                 


Val Leu Tyr Asp Ile Ile Leu Lys Asn Phe Glu Lys Leu Lys Lys Pro 
225                 230                 235                 240 


Arg Gly Glu Thr Lys Gly Met Ile Tyr Gly Ser Ser Trp Ser His Pro 
                245                 250                 255     


Gln Phe Glu Lys 
            260 


<210>  11
<211>  11
<212>  PRT
<213>  Escherichia coli

<400>  11

Ala Ala Asn Asp Glu Asn Tyr Ala Leu Ala Ala 
1               5                   10      


<210>  12
<211>  123
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Substrate S1

<400>  12

Met Gly His His His His His His Ser Ser Arg Arg Arg Arg Arg Arg 
1               5                   10                  15      


Arg Arg Arg Arg Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser 
            20                  25                  30          


Ser Ser Ser Phe Gly Tyr Gly Trp Ser Ser Ser Ser Ser Ser Ser Ser 
        35                  40                  45              


Ser Ser Ser Ser Ser Ser Ser Arg Arg Arg Arg Arg Arg Arg Arg Arg 
    50                  55                  60                  


Arg Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser 
65                  70                  75                  80  


Phe Gly Tyr Gly Trp Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser 
                85                  90                  95      


Ser Ser Ser Ser Arg Arg Arg Arg Arg Arg Arg Arg Arg Arg Ser Ser 
            100                 105                 110         


Ala Ala Asn Asp Glu Asn Tyr Ala Leu Ala Ala 
        115                 120             


<210>  13
<211>  210
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Substrate S2

<400>  13

Met Gly His His His His His His Ser Ser Arg Arg Arg Arg Arg Pro 
1               5                   10                  15      


Val Pro Leu Pro Ile Pro Val Pro Leu Pro Ile Pro Val Pro Leu Pro 
            20                  25                  30          


Ile Pro Arg Arg Arg Arg Arg Ser Ser Ser Ser Ser Ser Ser Ser Ser 
        35                  40                  45              


Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser 
    50                  55                  60                  


Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser 
65                  70                  75                  80  


Ser Ser Ser Ser Ser Ser Ser Ser Ser Arg Arg Arg Arg Arg Pro Val 
                85                  90                  95      


Pro Leu Pro Ile Pro Val Pro Leu Pro Ile Pro Val Pro Leu Pro Ile 
            100                 105                 110         


Pro Arg Arg Arg Arg Arg Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser 
        115                 120                 125             


Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser 
    130                 135                 140                 


Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser 
145                 150                 155                 160 


Ser Ser Ser Ser Ser Ser Ser Ser Glu Glu Glu Glu Glu Pro Val Pro 
                165                 170                 175     


Leu Pro Ile Pro Val Pro Leu Pro Ile Pro Val Pro Leu Pro Ile Pro 
            180                 185                 190         


Glu Glu Glu Glu Glu Ser Ser Ala Ala Asn Asp Glu Asn Tyr Ala Leu 
        195                 200                 205             


Ala Ala 
    210 


<210>  14
<211>  260
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Substrate S3

<400>  14

Met Gly His His His His His His Ser Ser Ser Lys Gly Glu Glu Leu 
1               5                   10                  15      


Phe Thr Gly Val Val Pro Ile Leu Val Glu Leu Asp Gly Asp Val Asn 
            20                  25                  30          


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


Gly Lys Leu Thr Leu Lys Phe Ile Cys Thr Thr Gly Lys Leu Pro Val 
    50                  55                  60                  


Pro Trp Pro Thr Leu Val Thr Thr Leu Thr Tyr Gly Val Gln Cys Phe 
65                  70                  75                  80  


Ser Arg Tyr Pro Asp His Met Lys Arg His Asp Phe Phe Lys Ser Ala 
                85                  90                  95      


Met Pro Glu Gly Tyr Val Gln Glu Arg Thr Ile Ser Phe Lys Asp Asp 
            100                 105                 110         


Gly Asn Tyr Lys Thr Arg Ala Glu Val Lys Phe Glu Gly Asp Thr Leu 
        115                 120                 125             


Val Asn Arg Ile Glu Leu Lys Gly Ile Asp Phe Lys Glu Asp Gly Asn 
    130                 135                 140                 


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


Ile Thr Ala Asp Lys Gln Lys Asn Gly Ile Lys Ala Asn Phe Lys Ile 
                165                 170                 175     


Arg His Asn Ile Glu Asp Gly Ser Val Gln Leu Ala Asp His Tyr Gln 
            180                 185                 190         


Gln Asn Thr Pro Ile Gly Asp Gly Pro Val Leu Leu Pro Asp Asn His 
        195                 200                 205             


Tyr Leu Ser Thr Gln Ser Ala Leu Ser Lys Asp Pro Asn Glu Lys Arg 
    210                 215                 220                 


Asp His Met Val Leu Leu Glu Phe Val Thr Ala Ala Gly Ile Thr His 
225                 230                 235                 240 


Gly Met Asp Glu Leu Tyr Lys Ser Ser Ala Ala Asn Asp Glu Asn Tyr 
                245                 250                 255     


Ala Leu Ala Ala 
            260 


<210>  15
<211>  5
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Linker 1

<400>  15

Gly Ser Ser Ser Glu 
1               5   


<210>  16
<211>  5
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Linker 2

<400>  16

Ser Asn Ser Ser Gly 
1               5   


<210>  17
<211>  32
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  TM sequence with linkers

<400>  17

Gly Ser Ser Ser Glu Val His Gly Asn Ala Glu Val His Ala Ser Phe 
1               5                   10                  15      


Phe Asp Ile Gly Gly Ser Val Ser Ala Gly Phe Ser Asn Ser Ser Gly 
            20                  25                  30          


<210>  18
<211>  12
<212>  PRT
<213>  Thermoplasma acidophilum

<400>  18

Ser Val Asp Ala Glu Ser Gly Lys Thr Lys Gly Gly 
1               5                   10          


<210>  19
<211>  263
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  N-terminally Strep-tagged subunit artificial PA26-nanopore

<400>  19

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


Arg Ala Ala Leu Ile Gln Asn Leu Arg Asp Ser Tyr Thr Glu Thr Ser 
            20                  25                  30          


Ser Phe Ala Val Ile Glu Glu Trp Ala Ala Gly Thr Leu Gln Glu Ile 
        35                  40                  45              


Glu Gly Ile Ala Lys Ala Ala Ala Glu Ala His Gly Val Ile Arg Asn 
    50                  55                  60                  


Ser Thr Tyr Gly Arg Ala Gln Ala Glu Lys Ser Pro Glu Gln Leu Leu 
65                  70                  75                  80  


Gly Val Leu Gln Arg Tyr Gln Asp Leu Cys His Asn Val Tyr Cys Gln 
                85                  90                  95      


Ala Glu Thr Ile Arg Thr Val Ile Ala Ile Arg Ile Pro Glu His Lys 
            100                 105                 110         


Glu Glu Asp Asn Leu Gly Val Ala Val Gln His Ala Val Leu Lys Ile 
        115                 120                 125             


Ile Asp Glu Leu Glu Ile Lys Thr Leu Gly Ser Gly Glu Lys Ser Gly 
    130                 135                 140                 


Ser Gly Gly Ala Pro Thr Pro Ile Gly Met Tyr Ala Leu Arg Glu Tyr 
145                 150                 155                 160 


Leu Ser Ala Arg Ser Thr Val Glu Asp Lys Leu Leu Gly Gly Ser Ser 
                165                 170                 175     


Ser Glu Val His Gly Asn Ala Glu Val His Ala Ser Phe Phe Asp Ile 
            180                 185                 190         


Gly Gly Ser Val Ser Ala Gly Phe Ser Asn Ser Ser Gly Ser Gln Ser 
        195                 200                 205             


Pro Ser Leu Leu Leu Glu Leu Arg Gln Ile Asp Ala Asp Phe Met Leu 
    210                 215                 220                 


Lys Val Glu Leu Ala Thr Thr His Leu Ser Thr Met Val Arg Ala Val 
225                 230                 235                 240 


Ile Asn Ala Tyr Leu Leu Asn Trp Lys Lys Leu Ile Gln Pro Arg Thr 
                245                 250                 255     


Gly Ser Asp His Met Val Ser 
            260             


<210>  20
<211>  243
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  N-term S-tagged PA26 nanopore with del seq

<400>  20

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


Arg Ala Ala Leu Ile Gln Asn Leu Arg Asp Ser Tyr Thr Glu Thr Ser 
            20                  25                  30          


Ser Phe Ala Val Ile Glu Glu Trp Ala Ala Gly Thr Leu Gln Glu Ile 
        35                  40                  45              


Glu Gly Ile Ala Lys Ala Ala Ala Glu Ala His Gly Val Ile Arg Asn 
    50                  55                  60                  


Ser Thr Tyr Gly Arg Ala Gln Ala Glu Lys Ser Pro Glu Gln Leu Leu 
65                  70                  75                  80  


Gly Val Leu Gln Arg Tyr Gln Asp Leu Cys His Asn Val Tyr Cys Gln 
                85                  90                  95      


Ala Glu Thr Ile Arg Thr Val Ile Ala Ile Arg Ile Pro Glu His Lys 
            100                 105                 110         


Glu Glu Asp Asn Leu Gly Val Ala Val Gln His Ala Val Leu Lys Ile 
        115                 120                 125             


Ile Asp Glu Leu Glu Ile Lys Thr Leu Gly Ser Gly Glu Lys Ser Gly 
    130                 135                 140                 


Ser Gly Gly Ala Pro Thr Pro Ile Gly Met Tyr Ala Leu Arg Glu Tyr 
145                 150                 155                 160 


Leu Ser Ala Arg Ser Thr Val Glu Asp Lys Leu Leu Gly Ser Val Asp 
                165                 170                 175     


Ala Glu Ser Gly Lys Thr Lys Gly Gly Ser Gln Ser Pro Ser Leu Leu 
            180                 185                 190         


Leu Glu Leu Arg Gln Ile Asp Ala Asp Phe Met Leu Lys Val Glu Leu 
        195                 200                 205             


Ala Thr Thr His Leu Ser Thr Met Val Arg Ala Val Ile Asn Ala Tyr 
    210                 215                 220                 


Leu Leu Asn Trp Lys Lys Leu Ile Gln Pro Arg Thr Gly Ser Asp His 
225                 230                 235                 240 


Met Val Ser 
            


<210>  21
<211>  5
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  linker

<400>  21

Ser Ser Ser Ser Ser 
1               5   


<210>  22
<211>  32
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  TM sequence with linkers

<400>  22

Ser Ser Ser Ser Ser Val His Gly Asn Ala Glu Val His Ala Ser Phe 
1               5                   10                  15      


Phe Asp Ile Gly Gly Ser Val Ser Ala Gly Phe Ser Ser Ser Ser Ser 
            20                  25                  30          


<210>  23
<211>  9
<212>  PRT
<213>  Aquifex aeolicus

<400>  23

Lys Gly Ala Phe Thr Gly Ala Val Ser 
1               5                   


<210>  24
<211>  302
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  N-term S-tag artificial ATPase with TM sequence

<400>  24

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


Asn Glu Leu Leu Arg Arg Glu Lys Asp Leu Lys Glu Glu Glu Tyr Val 
            20                  25                  30          


Phe Glu Ser Pro Lys Met Lys Glu Ile Leu Glu Lys Ile Lys Lys Ile 
        35                  40                  45              


Ser Cys Ala Glu Cys Pro Val Leu Ile Thr Gly Glu Ser Gly Val Gly 
    50                  55                  60                  


Lys Glu Val Val Ala Arg Leu Ile His Lys Leu Ser Asp Arg Ser Lys 
65                  70                  75                  80  


Glu Pro Phe Val Ala Leu Asn Val Ala Ser Ile Pro Arg Asp Ile Phe 
                85                  90                  95      


Glu Ala Glu Leu Phe Gly Tyr Glu Ser Ser Ser Ser Ser Val His Gly 
            100                 105                 110         


Asn Ala Glu Val His Ala Ser Phe Phe Asp Ile Gly Gly Ser Val Ser 
        115                 120                 125             


Ala Gly Phe Ser Ser Ser Ser Ser Ser Lys Glu Gly Phe Phe Glu Leu 
    130                 135                 140                 


Ala Asp Gly Gly Thr Leu Phe Leu Asp Ala Ile Gly Glu Leu Ser Leu 
145                 150                 155                 160 


Glu Ala Gln Ala Lys Leu Leu Arg Val Ile Glu Ser Gly Lys Phe Tyr 
                165                 170                 175     


Arg Leu Gly Gly Arg Lys Glu Ile Glu Val Asn Val Arg Ile Leu Ala 
            180                 185                 190         


Ala Thr Asn Arg Asn Ile Lys Glu Leu Val Lys Glu Gly Lys Phe Arg 
        195                 200                 205             


Glu Asp Leu Tyr Tyr Arg Leu Gly Val Ile Glu Ile Glu Ile Pro Pro 
    210                 215                 220                 


Leu Arg Glu Arg Lys Glu Asp Ile Ile Pro Leu Ala Asn His Phe Leu 
225                 230                 235                 240 


Lys Lys Phe Ser Arg Lys Tyr Ala Lys Glu Val Glu Gly Phe Thr Lys 
                245                 250                 255     


Ser Ala Gln Glu Leu Leu Leu Ser Tyr Pro Trp Tyr Gly Asn Val Arg 
            260                 265                 270         


Glu Leu Lys Asn Val Ile Glu Arg Ala Val Leu Phe Ser Glu Gly Lys 
        275                 280                 285             


Phe Ile Asp Arg Gly Glu Leu Ser Cys Leu Val Asn Ser Lys 
    290                 295                 300         


<210>  25
<211>  279
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  N-term S-tag artificial ATPase nanopore

<400>  25

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


Asn Glu Leu Leu Arg Arg Glu Lys Asp Leu Lys Glu Glu Glu Tyr Val 
            20                  25                  30          


Phe Glu Ser Pro Lys Met Lys Glu Ile Leu Glu Lys Ile Lys Lys Ile 
        35                  40                  45              


Ser Cys Ala Glu Cys Pro Val Leu Ile Thr Gly Glu Ser Gly Val Gly 
    50                  55                  60                  


Lys Glu Val Val Ala Arg Leu Ile His Lys Leu Ser Asp Arg Ser Lys 
65                  70                  75                  80  


Glu Pro Phe Val Ala Leu Asn Val Ala Ser Ile Pro Arg Asp Ile Phe 
                85                  90                  95      


Glu Ala Glu Leu Phe Gly Tyr Glu Lys Gly Ala Phe Thr Gly Ala Val 
            100                 105                 110         


Ser Ser Lys Glu Gly Phe Phe Glu Leu Ala Asp Gly Gly Thr Leu Phe 
        115                 120                 125             


Leu Asp Ala Ile Gly Glu Leu Ser Leu Glu Ala Gln Ala Lys Leu Leu 
    130                 135                 140                 


Arg Val Ile Glu Ser Gly Lys Phe Tyr Arg Leu Gly Gly Arg Lys Glu 
145                 150                 155                 160 


Ile Glu Val Asn Val Arg Ile Leu Ala Ala Thr Asn Arg Asn Ile Lys 
                165                 170                 175     


Glu Leu Val Lys Glu Gly Lys Phe Arg Glu Asp Leu Tyr Tyr Arg Leu 
            180                 185                 190         


Gly Val Ile Glu Ile Glu Ile Pro Pro Leu Arg Glu Arg Lys Glu Asp 
        195                 200                 205             


Ile Ile Pro Leu Ala Asn His Phe Leu Lys Lys Phe Ser Arg Lys Tyr 
    210                 215                 220                 


Ala Lys Glu Val Glu Gly Phe Thr Lys Ser Ala Gln Glu Leu Leu Leu 
225                 230                 235                 240 


Ser Tyr Pro Trp Tyr Gly Asn Val Arg Glu Leu Lys Asn Val Ile Glu 
                245                 250                 255     


Arg Ala Val Leu Phe Ser Glu Gly Lys Phe Ile Asp Arg Gly Glu Leu 
            260                 265                 270         


Ser Cys Leu Val Asn Ser Lys 
        275                 


<210>  26
<211>  462
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  C-terminally Strep tagged subunit of artificial ClpP-nanopore

<400>  26

Met Gly Ser Tyr Ser Gly Glu Arg Asp Asn Phe Ala Pro His Met Ala 
1               5                   10                  15      


Leu Val Pro Met Val Ile Glu Gln Thr Ser Arg Gly Glu Arg Ser Phe 
            20                  25                  30          


Asp Ile Tyr Ser Arg Leu Leu Lys Glu Arg Val Ile Phe Leu Thr Gly 
        35                  40                  45              


Gln Val Glu Asp His Met Ala Asn Leu Ile Val Ala Gln Met Leu Phe 
    50                  55                  60                  


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


Pro Gly Gly Val Ile Thr Ala Gly Met Ser Ile Tyr Asp Thr Met Gln 
                85                  90                  95      


Phe Ile Lys Pro Asp Val Ser Thr Ile Cys Met Gly Gln Ala Ala Ser 
            100                 105                 110         


Met Gly Ala Phe Leu Leu Thr Ala Gly Ala Lys Gly Lys Arg Phe Cys 
        115                 120                 125             


Leu Pro Asn Ser Arg Val Met Ile His Gln Pro Leu Gly Gly Tyr Gln 
    130                 135                 140                 


Gly Gln Ala Thr Asp Ile Glu Ile His Ala Arg Glu Ile Leu Lys Val 
145                 150                 155                 160 


Lys Gly Arg Met Asn Glu Leu Met Ala Leu His Thr Gly Gln Ser Leu 
                165                 170                 175     


Glu Gln Ile Glu Arg Asp Thr Glu Arg Asp Arg Phe Leu Ser Ala Pro 
            180                 185                 190         


Glu Ala Val Glu Tyr Gly Leu Val Asp Ser Ile Leu Thr His Arg Asn 
        195                 200                 205             


Ala Thr Leu Arg Val His Pro Glu Ala Gln Ala Lys Val Asp Val Phe 
    210                 215                 220                 


Arg Glu Asp Leu Cys Ser Lys Thr Glu Asn Leu Leu Gly Ser Tyr Phe 
225                 230                 235                 240 


Pro Lys Lys Ile Ser Glu Leu Asp Ala Phe Leu Lys Glu Pro Ala Leu 
                245                 250                 255     


Asn Glu Ala Asn Leu Ser Asn Leu Lys Ala Pro Leu Asp Ile Gly Ser 
            260                 265                 270         


Ser Ser Glu Val His Gly Asn Ala Glu Val His Ala Ser Phe Phe Asp 
        275                 280                 285             


Ile Gly Gly Ser Val Ser Ala Gly Phe Ser Asn Ser Ser Gly Cys Gly 
    290                 295                 300                 


Pro Val Asn Cys Asn Glu Lys Ile Val Val Leu Leu Gln Arg Leu Lys 
305                 310                 315                 320 


Pro Glu Ile Lys Asp Val Thr Glu Gln Leu Asn Leu Val Thr Thr Trp 
                325                 330                 335     


Leu Gln Leu Gln Ile Pro Arg Ile Glu Asp Gly Asn Asn Phe Gly Val 
            340                 345                 350         


Ala Val Gln Glu Lys Val Phe Glu Leu Met Thr Asn Leu His Thr Lys 
        355                 360                 365             


Leu Glu Gly Phe His Thr Gln Ile Ser Lys Tyr Phe Ser Glu Arg Gly 
    370                 375                 380                 


Asp Ala Val Ala Lys Ala Ala Lys Gln Pro His Val Gly Asp Tyr Arg 
385                 390                 395                 400 


Gln Leu Val His Glu Leu Asp Glu Ala Glu Tyr Gln Glu Ile Arg Leu 
                405                 410                 415     


Met Val Met Glu Ile Arg Asn Ala Tyr Ala Val Leu Tyr Asp Ile Ile 
            420                 425                 430         


Leu Lys Asn Phe Glu Lys Leu Lys Lys Pro Arg Gly Glu Thr Lys Gly 
        435                 440                 445             


Met Ile Tyr Gly Ser Ser Trp Ser His Pro Gln Phe Glu Lys 
    450                 455                 460         


<210>  27
<211>  30
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  TM region beta strand

<400>  27

Phe Asp Ile Gly Gly Ser Val Ser Ala Gly Phe Ser Asn Ser Asn Ser 
1               5                   10                  15      


Ser Thr Val Ala Ile Asp His Ser Leu Ser Leu Ser Ser Gly 
            20                  25                  30  


<210>  28
<211>  30
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  TM region beta strand

<400>  28

Gly Ser Ser Arg Thr Ile Ser Lys Asn Thr Ser Thr Ser Arg Thr His 
1               5                   10                  15      


Thr Ser Glu Val His Gly Asn Ala Glu Val His Ala Ser Phe 
            20                  25                  30  


<210>  29
<211>  7
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Linker sequence d4 mutant

<400>  29

Gly Ser Ser His Thr Ser Glu 
1               5           


<210>  30
<211>  7
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Linker sequence d4 mutant

<400>  30

Ser Asn Ser Asn Ser Ser Gly 
1               5           


<210>  31
<211>  5
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  linker sequence d2 mutant

<400>  31

Gly Ser Ser Ser Glu 
1               5   


<210>  32
<211>  5
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  linker sequence d2 mutant

<400>  32

Ser Asn Ser Ser Gly 
1               5   


<210>  33
<211>  10
<212>  PRT
<213>  Homo sapiens

<400>  33

Asp Arg Val Tyr Ile His Pro Phe His Leu 
1               5                   10  


<210>  34
<211>  17
<212>  PRT
<213>  Homo sapiens

<400>  34

Tyr Gly Gly Phe Leu Arg Arg Ile Arg Pro Lys Leu Lys Trp Asp Asn 
1               5                   10                  15      


Gln 
    


<210>  35
<211>  7
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  part of S2

<400>  35

Pro Val Pro Leu Pro Ile Pro 
1               5           


<210>  36
<211>  19
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  part of substrate 2

<400>  36

Pro Val Pro Leu Pro Ile Pro Val Pro Leu Pro Ile Pro Val Pro Leu 
1               5                   10                  15      


Pro Ile Pro 
            


