                         SEQUENCE LISTING

<110>  University of Washington
       Nivala, Jeffrey Matthew
 
<120>  REPORTER CONSTRUCTS FOR NANOPORE-BASED DETECTION OF BIOLOGICAL 
       ACTIVITY

<130>  UWOTL170215

<150>  US 62/741,670
<151>  2018-10-05

<160>  36    

<170>  PatentIn version 3.5

<210>  1
<211>  389
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic


<220>
<221>  misc_feature
<222>  (312)..(342)
<223>  Xaa can be any naturally occurring amino acid

<400>  1

Met Thr Met Thr Arg Leu Lys Ile Ser Lys Thr Leu Leu Ala Val Met 
1               5                   10                  15      


Leu Thr Ser Ala Val Ala Thr Gly Ser Ala Tyr Ala Glu Asn Asn Ala 
            20                  25                  30          


Gln Thr Thr Asn Glu Ser Ala Gly Gln Lys Val Asp Ser Ser Met Asn 
        35                  40                  45              


Lys Val Gly Asn Phe Met Asp Asp Ser Ala Ile Thr Ala Lys Val Lys 
    50                  55                  60                  


Ala Ala Leu Val Asp His Asp Asn Ile Lys Ser Thr Asp Ile Ser Val 
65                  70                  75                  80  


Lys Thr Asp Gln Lys Val Val Thr Leu Ser Gly Phe Val Glu Ser Gln 
                85                  90                  95      


Ala Gln Ala Glu Glu Ala Val Lys Val Ala Lys Gly Val Glu Gly Val 
            100                 105                 110         


Thr Ser Val Ser Asp Lys Leu His Val Arg Asp Ala Lys Glu Gly Ser 
        115                 120                 125             


Val Lys Gly Tyr Ala Gly Asp Thr Ala Thr Thr Ser Glu Ile Lys Ala 
    130                 135                 140                 


Lys Leu Leu Ala Asp Asp Ile Val Pro Ser Arg His Val Lys Val Glu 
145                 150                 155                 160 


Thr Thr Asp Gly Val Val Gln Leu Ser Gly Thr Val Asp Ser Gln Ala 
                165                 170                 175     


Gln Ser Asp Arg Ala Glu Ser Ile Ala Lys Ala Val Asp Gly Val Lys 
            180                 185                 190         


Ser Val Lys Asn Asp Leu Lys Thr Lys Met Gly His His His His His 
        195                 200                 205             


His His His His His Gly Ser Leu Gln Asp Ser Glu Val Asn Gln Glu 
    210                 215                 220                 


Ala Lys Pro Glu Val Lys Pro Glu Val Lys Pro Glu Thr His Ile Asn 
225                 230                 235                 240 


Leu Lys Val Ser Asp Gly Ser Ser Glu Ile Phe Phe Lys Ile Lys Lys 
                245                 250                 255     


Thr Thr Pro Leu Arg Arg Leu Met Glu Ala Phe Ala Lys Arg Gln Gly 
            260                 265                 270         


Lys Glu Met Asp Ser Leu Arg Phe Leu Tyr Asp Gly Ile Arg Ile Gln 
        275                 280                 285             


Ala Asp Gln Ala Pro Glu Asp Leu Asp Met Glu Asp Asn Asp Ile Ile 
    290                 295                 300                 


Glu Ala His Arg Glu Gln Ile Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 
305                 310                 315                 320 


Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 
                325                 330                 335     


Xaa Xaa Xaa Xaa Xaa Xaa Asp Gly Gly Ser Ser Gly Gly Ser Gly Gly 
            340                 345                 350         


Asp Gly Ser Ser Gly Asp Gly Gly Ser Asp Gly Asp Ser Asp Gly Ser 
        355                 360                 365             


Asp Gly Asp Gly Asp Ser Asp Gly Asp Asp Gly Gly Asp Asp Glu Asp 
    370                 375                 380                 


Asp Gly Ser Asp Asp 
385                 


<210>  2
<211>  31
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic

<400>  2

Gly Gly Gly Gly Ser Ser Gly Gly Ser Gly Gly Ser Gly Ser Ser Gly 
1               5                   10                  15      


Asp Gly Gly Ser Ser Gly Gly Ser Gly Gly Ser Gly Ser Ser Gly 
            20                  25                  30      


<210>  3
<211>  31
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic

<400>  3

Tyr Tyr Tyr Gly Ser Ser Gly Gly Ser Gly Gly Ser Gly Ser Ser Gly 
1               5                   10                  15      


Asp Gly Gly Ser Ser Gly Gly Ser Gly Gly Ser Gly Ser Ser Gly 
            20                  25                  30      


<210>  4
<211>  31
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic

<400>  4

Gly Gly Tyr Tyr Tyr Ser Gly Gly Ser Gly Gly Ser Gly Ser Ser Gly 
1               5                   10                  15      


Asp Gly Gly Ser Ser Gly Gly Ser Gly Gly Ser Gly Ser Ser Gly 
            20                  25                  30      


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

<220>
<223>  Synthetic

<400>  5

Gly Gly Gly Gly Tyr Tyr Tyr Gly Ser Gly Gly Ser Gly Ser Ser Gly 
1               5                   10                  15      


Asp Gly Gly Ser Ser Gly Gly Ser Gly Gly Ser Gly Ser Ser Gly 
            20                  25                  30      


<210>  6
<211>  31
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic

<400>  6

Gly Gly Gly Gly Ser Ser Tyr Tyr Tyr Gly Gly Ser Gly Ser Ser Gly 
1               5                   10                  15      


Asp Gly Gly Ser Ser Gly Gly Ser Gly Gly Ser Gly Ser Ser Gly 
            20                  25                  30      


<210>  7
<211>  31
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic

<400>  7

Gly Gly Gly Gly Ser Ser Gly Gly Tyr Tyr Tyr Ser Gly Ser Ser Gly 
1               5                   10                  15      


Asp Gly Gly Ser Ser Gly Gly Ser Gly Gly Ser Gly Ser Ser Gly 
            20                  25                  30      


<210>  8
<211>  31
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic

<400>  8

Gly Gly Gly Gly Ser Ser Gly Gly Ser Gly Tyr Tyr Tyr Ser Ser Gly 
1               5                   10                  15      


Asp Gly Gly Ser Ser Gly Gly Ser Gly Gly Ser Gly Ser Ser Gly 
            20                  25                  30      


<210>  9
<211>  31
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic

<400>  9

Gly Gly Gly Gly Ser Ser Gly Gly Ser Gly Gly Ser Tyr Tyr Tyr Gly 
1               5                   10                  15      


Asp Gly Gly Ser Ser Gly Gly Ser Gly Gly Ser Gly Ser Ser Gly 
            20                  25                  30      


<210>  10
<211>  31
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic

<400>  10

Gly Gly Gly Gly Ser Ser Gly Gly Ser Gly Gly Ser Gly Ser Tyr Tyr 
1               5                   10                  15      


Asp Gly Gly Ser Ser Gly Gly Ser Gly Gly Ser Gly Ser Ser Gly 
            20                  25                  30      


<210>  11
<211>  31
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic

<400>  11

Gly Gly Gly Gly Ser Ser Gly Gly Ser Gly Gly Ser Gly Ser Ser Gly 
1               5                   10                  15      


Asp Tyr Tyr Ser Ser Gly Gly Ser Gly Gly Ser Gly Ser Ser Gly 
            20                  25                  30      


<210>  12
<211>  31
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic

<400>  12

Gly Gly Gly Gly Ser Ser Gly Gly Ser Gly Gly Ser Gly Ser Ser Gly 
1               5                   10                  15      


Asp Gly Tyr Tyr Tyr Gly Gly Ser Gly Gly Ser Gly Ser Ser Gly 
            20                  25                  30      


<210>  13
<211>  31
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic

<400>  13

Gly Gly Gly Gly Ser Ser Gly Gly Ser Gly Gly Ser Gly Ser Ser Gly 
1               5                   10                  15      


Asp Gly Gly Ser Tyr Tyr Tyr Ser Gly Gly Ser Gly Ser Ser Gly 
            20                  25                  30      


<210>  14
<211>  31
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic

<400>  14

Gly Gly Gly Gly Ser Ser Gly Gly Ser Gly Gly Ser Gly Ser Ser Gly 
1               5                   10                  15      


Asp Gly Gly Ser Ser Gly Tyr Tyr Tyr Gly Ser Gly Ser Ser Gly 
            20                  25                  30      


<210>  15
<211>  31
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic

<400>  15

Gly Gly Gly Gly Ser Ser Gly Gly Ser Gly Gly Ser Gly Ser Ser Gly 
1               5                   10                  15      


Asp Gly Gly Ser Ser Gly Gly Ser Tyr Tyr Tyr Gly Ser Ser Gly 
            20                  25                  30      


<210>  16
<211>  31
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic

<400>  16

Gly Gly Gly Gly Ser Ser Gly Gly Ser Gly Gly Ser Gly Ser Ser Gly 
1               5                   10                  15      


Asp Gly Gly Ser Ser Gly Gly Ser Gly Gly Tyr Tyr Tyr Ser Gly 
            20                  25                  30      


<210>  17
<211>  31
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic

<400>  17

Gly Gly Gly Gly Ser Ser Gly Gly Ser Gly Gly Ser Gly Ser Ser Gly 
1               5                   10                  15      


Asp Gly Gly Ser Ser Gly Gly Ser Gly Gly Ser Gly Tyr Tyr Tyr 
            20                  25                  30      


<210>  18
<211>  31
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic

<400>  18

Gly Gly Ala Ala Ala Ala Ala Ala Ala Ala Ala Ser Gly Ser Ser Gly 
1               5                   10                  15      


Asp Gly Gly Ser Ser Gly Gly Ser Gly Gly Ser Gly Ser Ser Gly 
            20                  25                  30      


<210>  19
<211>  31
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic

<400>  19

Gly Gly Asp Asp Asp Asp Asp Asp Asp Asp Asp Ser Gly Ser Ser Gly 
1               5                   10                  15      


Asp Gly Gly Ser Ser Gly Gly Ser Gly Gly Ser Gly Ser Ser Gly 
            20                  25                  30      


<210>  20
<211>  31
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic

<400>  20

Gly Gly Glu Glu Glu Glu Glu Glu Glu Glu Glu Ser Gly Ser Ser Gly 
1               5                   10                  15      


Asp Gly Gly Ser Ser Gly Gly Ser Gly Gly Ser Gly Ser Ser Gly 
            20                  25                  30      


<210>  21
<211>  31
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic

<400>  21

Gly Gly Gly Gly Gly Gly Gly Gly Gly Gly Gly Ser Gly Ser Ser Gly 
1               5                   10                  15      


Asp Gly Gly Ser Ser Gly Gly Ser Gly Gly Ser Gly Ser Ser Gly 
            20                  25                  30      


<210>  22
<211>  31
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic

<400>  22

Gly Gly His His His His His His His His His Ser Gly Ser Ser Gly 
1               5                   10                  15      


Asp Gly Gly Ser Ser Gly Gly Ser Gly Gly Ser Gly Ser Ser Gly 
            20                  25                  30      


<210>  23
<211>  31
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic

<400>  23

Gly Gly Met Met Met Met Met Met Met Met Met Ser Gly Ser Ser Gly 
1               5                   10                  15      


Asp Gly Gly Ser Ser Gly Gly Ser Gly Gly Ser Gly Ser Ser Gly 
            20                  25                  30      


<210>  24
<211>  31
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic

<400>  24

Gly Gly Asn Asn Asn Asn Asn Asn Asn Asn Asn Ser Gly Ser Ser Gly 
1               5                   10                  15      


Asp Gly Gly Ser Ser Gly Gly Ser Gly Gly Ser Gly Ser Ser Gly 
            20                  25                  30      


<210>  25
<211>  31
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic

<400>  25

Gly Gly Pro Pro Pro Pro Pro Pro Pro Pro Pro Ser Gly Ser Ser Gly 
1               5                   10                  15      


Asp Gly Gly Ser Ser Gly Gly Ser Gly Gly Ser Gly Ser Ser Gly 
            20                  25                  30      


<210>  26
<211>  31
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic

<400>  26

Gly Gly Gln Gln Gln Gln Gln Gln Gln Gln Gln Ser Gly Ser Ser Gly 
1               5                   10                  15      


Asp Gly Gly Ser Ser Gly Gly Ser Gly Gly Ser Gly Ser Ser Gly 
            20                  25                  30      


<210>  27
<211>  31
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic

<400>  27

Gly Gly Arg Arg Arg Arg Arg Arg Arg Arg Arg Ser Gly Ser Ser Gly 
1               5                   10                  15      


Asp Gly Gly Ser Ser Gly Gly Ser Gly Gly Ser Gly Ser Ser Gly 
            20                  25                  30      


<210>  28
<211>  31
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic

<400>  28

Gly Gly Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Gly Ser Ser Gly 
1               5                   10                  15      


Asp Gly Gly Ser Ser Gly Gly Ser Gly Gly Ser Gly Ser Ser Gly 
            20                  25                  30      


<210>  29
<211>  31
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic

<400>  29

Gly Gly Thr Thr Thr Thr Thr Thr Thr Thr Thr Ser Gly Ser Ser Gly 
1               5                   10                  15      


Asp Gly Gly Ser Ser Gly Gly Ser Gly Gly Ser Gly Ser Ser Gly 
            20                  25                  30      


<210>  30
<211>  31
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic

<400>  30

Gly Gly Arg Arg Gly Ser Tyr Tyr Ser Gly Gly Ser Gly Ser Ser Gly 
1               5                   10                  15      


Asp Gly Gly Ser Ser Gly Gly Ser Gly Gly Ser Gly Ser Ser Gly 
            20                  25                  30      


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

<220>
<223>  Synthetic

<400>  31

Gly Gly Arg Arg Gly Glu Tyr Tyr Ser Gly Gly Ser Gly Ser Ser Gly 
1               5                   10                  15      


Asp Gly Gly Ser Ser Gly Gly Ser Gly Gly Ser Gly Ser Ser Gly 
            20                  25                  30      


<210>  32
<211>  201
<212>  PRT
<213>  Escherichia coli

<400>  32

Met Thr Met Thr Arg Leu Lys Ile Ser Lys Thr Leu Leu Ala Val Met 
1               5                   10                  15      


Leu Thr Ser Ala Val Ala Thr Gly Ser Ala Tyr Ala Glu Asn Asn Ala 
            20                  25                  30          


Gln Thr Thr Asn Glu Ser Ala Gly Gln Lys Val Asp Ser Ser Met Asn 
        35                  40                  45              


Lys Val Gly Asn Phe Met Asp Asp Ser Ala Ile Thr Ala Lys Val Lys 
    50                  55                  60                  


Ala Ala Leu Val Asp His Asp Asn Ile Lys Ser Thr Asp Ile Ser Val 
65                  70                  75                  80  


Lys Thr Asp Gln Lys Val Val Thr Leu Ser Gly Phe Val Glu Ser Gln 
                85                  90                  95      


Ala Gln Ala Glu Glu Ala Val Lys Val Ala Lys Gly Val Glu Gly Val 
            100                 105                 110         


Thr Ser Val Ser Asp Lys Leu His Val Arg Asp Ala Lys Glu Gly Ser 
        115                 120                 125             


Val Lys Gly Tyr Ala Gly Asp Thr Ala Thr Thr Ser Glu Ile Lys Ala 
    130                 135                 140                 


Lys Leu Leu Ala Asp Asp Ile Val Pro Ser Arg His Val Lys Val Glu 
145                 150                 155                 160 


Thr Thr Asp Gly Val Val Gln Leu Ser Gly Thr Val Asp Ser Gln Ala 
                165                 170                 175     


Gln Ser Asp Arg Ala Glu Ser Ile Ala Lys Ala Val Asp Gly Val Lys 
            180                 185                 190         


Ser Val Lys Asn Asp Leu Lys Thr Lys 
        195                 200     


<210>  33
<211>  14
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic

<400>  33

Met Gly His His His His His His His His His His Gly Ser 
1               5                   10                  


<210>  34
<211>  96
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic

<400>  34

Leu Gln Asp Ser Glu Val Asn Gln Glu Ala Lys Pro Glu Val Lys Pro 
1               5                   10                  15      


Glu Val Lys Pro Glu Thr His Ile Asn Leu Lys Val Ser Asp Gly Ser 
            20                  25                  30          


Ser Glu Ile Phe Phe Lys Ile Lys Lys Thr Thr Pro Leu Arg Arg Leu 
        35                  40                  45              


Met Glu Ala Phe Ala Lys Arg Gln Gly Lys Glu Met Asp Ser Leu Arg 
    50                  55                  60                  


Phe Leu Tyr Asp Gly Ile Arg Ile Gln Ala Asp Gln Ala Pro Glu Asp 
65                  70                  75                  80  


Leu Asp Met Glu Asp Asn Asp Ile Ile Glu Ala His Arg Glu Gln Ile 
                85                  90                  95      


<210>  35
<211>  47
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic

<400>  35

Asp Gly Gly Ser Ser Gly Gly Ser Gly Gly Asp Gly Ser Ser Gly Asp 
1               5                   10                  15      


Gly Gly Ser Asp Gly Asp Ser Asp Gly Ser Asp Gly Asp Gly Asp Ser 
            20                  25                  30          


Asp Gly Asp Asp Gly Gly Asp Asp Glu Asp Asp Gly Ser Asp Asp 
        35                  40                  45          


<210>  36
<211>  118
<212>  PRT
<213>  Escherichia coli

<400>  36

Met Lys Lys Arg Gly Ala Phe Leu Gly Leu Leu Leu Val Ser Ala Cys 
1               5                   10                  15      


Ala Ser Val Phe Ala Ala Asn Asn Glu Thr Ser Lys Ser Val Thr Phe 
            20                  25                  30          


Pro Lys Cys Glu Asp Leu Asp Ala Ala Gly Ile Ala Ala Ser Val Lys 
        35                  40                  45              


Arg Asp Tyr Gln Gln Asn Arg Val Ala Arg Trp Ala Asp Asp Gln Lys 
    50                  55                  60                  


Ile Val Gly Gln Ala Asp Pro Val Ala Trp Val Ser Leu Gln Asp Ile 
65                  70                  75                  80  


Gln Gly Lys Asp Asp Lys Trp Ser Val Pro Leu Thr Val Arg Gly Lys 
                85                  90                  95      


Ser Ala Asp Ile His Tyr Gln Val Ser Val Asp Cys Lys Ala Gly Met 
            100                 105                 110         


Ala Glu Tyr Gln Arg Arg 
        115             


