                         SEQUENCE LISTING

<110>  CureVac AG
 
<120>  Immobilized inorganic pyrophosphatase (PPase)

<130>  C 9935-WO / RN

<160>  22    

<170>  PatentIn version 3.5

<210>  1
<211>  176
<212>  PRT
<213>  Escherichia coli

<400>  1

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


Tyr Val Val Ile Glu Ile Pro Ala Asn Ala Asp Pro Ile Lys Tyr Glu 
            20                  25                  30          


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


Ala Met Phe Tyr Pro Cys Asn Tyr Gly Tyr Ile Asn His Thr Leu Ser 
    50                  55                  60                  


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


Gln Pro Gly Ser Val Ile Arg Cys Arg Pro Val Gly Val Leu Lys Met 
                85                  90                  95      


Thr Asp Glu Ala Gly Glu Asp Ala Lys Leu Val Ala Val Pro His Ser 
            100                 105                 110         


Lys Leu Ser Lys Glu Tyr Asp His Ile Lys Asp Val Asn Asp Leu Pro 
        115                 120                 125             


Glu Leu Leu Lys Ala Gln Ile Ala His Phe Phe Glu His Tyr Lys Asp 
    130                 135                 140                 


Leu Glu Lys Gly Lys Trp Val Lys Val Glu Gly Trp Glu Asn Ala Glu 
145                 150                 155                 160 


Ala Ala Lys Ala Glu Ile Val Ala Ser Phe Glu Arg Ala Lys Asn Lys 
                165                 170                 175     


<210>  2
<211>  192
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Mutated Escherichia coli PPase_ C54S,C88A,177(GGGGS)3C

<400>  2

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


Tyr Val Val Ile Glu Ile Pro Ala Asn Ala Asp Pro Ile Lys Tyr Glu 
            20                  25                  30          


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


Ala Met Phe Tyr Pro Ser Asn Tyr Gly Tyr Ile Asn His Thr Leu Ser 
    50                  55                  60                  


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


Gln Pro Gly Ser Val Ile Arg Ala Arg Pro Val Gly Val Leu Lys Met 
                85                  90                  95      


Thr Asp Glu Ala Gly Glu Asp Ala Lys Leu Val Ala Val Pro His Ser 
            100                 105                 110         


Lys Leu Ser Lys Glu Tyr Asp His Ile Lys Asp Val Asn Asp Leu Pro 
        115                 120                 125             


Glu Leu Leu Lys Ala Gln Ile Ala His Phe Phe Glu His Tyr Lys Asp 
    130                 135                 140                 


Leu Glu Lys Gly Lys Trp Val Lys Val Glu Gly Trp Glu Asn Ala Glu 
145                 150                 155                 160 


Ala Ala Lys Ala Glu Ile Val Ala Ser Phe Glu Arg Ala Lys Asn Lys 
                165                 170                 175     


Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Cys 
            180                 185                 190         


<210>  3
<211>  192
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Mutated Escherichia coli PPase_ C54A,C88A,177(GGGGS)3C

<400>  3

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


Tyr Val Val Ile Glu Ile Pro Ala Asn Ala Asp Pro Ile Lys Tyr Glu 
            20                  25                  30          


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


Ala Met Phe Tyr Pro Ala Asn Tyr Gly Tyr Ile Asn His Thr Leu Ser 
    50                  55                  60                  


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


Gln Pro Gly Ser Val Ile Arg Ala Arg Pro Val Gly Val Leu Lys Met 
                85                  90                  95      


Thr Asp Glu Ala Gly Glu Asp Ala Lys Leu Val Ala Val Pro His Ser 
            100                 105                 110         


Lys Leu Ser Lys Glu Tyr Asp His Ile Lys Asp Val Asn Asp Leu Pro 
        115                 120                 125             


Glu Leu Leu Lys Ala Gln Ile Ala His Phe Phe Glu His Tyr Lys Asp 
    130                 135                 140                 


Leu Glu Lys Gly Lys Trp Val Lys Val Glu Gly Trp Glu Asn Ala Glu 
145                 150                 155                 160 


Ala Ala Lys Ala Glu Ile Val Ala Ser Phe Glu Arg Ala Lys Asn Lys 
                165                 170                 175     


Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Cys 
            180                 185                 190         


<210>  4
<211>  192
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Mutated Escherichia coli PPase_ C54A,C88S,177(GGGGS)3C

<400>  4

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


Tyr Val Val Ile Glu Ile Pro Ala Asn Ala Asp Pro Ile Lys Tyr Glu 
            20                  25                  30          


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


Ala Met Phe Tyr Pro Ala Asn Tyr Gly Tyr Ile Asn His Thr Leu Ser 
    50                  55                  60                  


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


Gln Pro Gly Ser Val Ile Arg Ser Arg Pro Val Gly Val Leu Lys Met 
                85                  90                  95      


Thr Asp Glu Ala Gly Glu Asp Ala Lys Leu Val Ala Val Pro His Ser 
            100                 105                 110         


Lys Leu Ser Lys Glu Tyr Asp His Ile Lys Asp Val Asn Asp Leu Pro 
        115                 120                 125             


Glu Leu Leu Lys Ala Gln Ile Ala His Phe Phe Glu His Tyr Lys Asp 
    130                 135                 140                 


Leu Glu Lys Gly Lys Trp Val Lys Val Glu Gly Trp Glu Asn Ala Glu 
145                 150                 155                 160 


Ala Ala Lys Ala Glu Ile Val Ala Ser Phe Glu Arg Ala Lys Asn Lys 
                165                 170                 175     


Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Cys 
            180                 185                 190         


<210>  5
<211>  192
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Mutated Escherichia coli PPase_ C54S,C88S,177(GGGGS)3C

<400>  5

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


Tyr Val Val Ile Glu Ile Pro Ala Asn Ala Asp Pro Ile Lys Tyr Glu 
            20                  25                  30          


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


Ala Met Phe Tyr Pro Ser Asn Tyr Gly Tyr Ile Asn His Thr Leu Ser 
    50                  55                  60                  


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


Gln Pro Gly Ser Val Ile Arg Ser Arg Pro Val Gly Val Leu Lys Met 
                85                  90                  95      


Thr Asp Glu Ala Gly Glu Asp Ala Lys Leu Val Ala Val Pro His Ser 
            100                 105                 110         


Lys Leu Ser Lys Glu Tyr Asp His Ile Lys Asp Val Asn Asp Leu Pro 
        115                 120                 125             


Glu Leu Leu Lys Ala Gln Ile Ala His Phe Phe Glu His Tyr Lys Asp 
    130                 135                 140                 


Leu Glu Lys Gly Lys Trp Val Lys Val Glu Gly Trp Glu Asn Ala Glu 
145                 150                 155                 160 


Ala Ala Lys Ala Glu Ile Val Ala Ser Phe Glu Arg Ala Lys Asn Lys 
                165                 170                 175     


Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Cys 
            180                 185                 190         


<210>  6
<211>  177
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Mutated Escherichia coli PPase_ C54S,C88A,177(GGGGS)3C

<400>  6

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


Tyr Val Val Ile Glu Ile Pro Ala Asn Ala Asp Pro Ile Lys Tyr Glu 
            20                  25                  30          


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


Ala Met Phe Tyr Pro Ser Asn Tyr Gly Tyr Ile Asn His Thr Leu Ser 
    50                  55                  60                  


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


Gln Pro Gly Ser Val Ile Arg Ala Arg Pro Val Gly Val Leu Lys Met 
                85                  90                  95      


Thr Asp Glu Ala Gly Glu Asp Ala Lys Leu Val Ala Val Pro His Ser 
            100                 105                 110         


Lys Leu Ser Lys Glu Tyr Asp His Ile Lys Asp Val Asn Asp Leu Pro 
        115                 120                 125             


Glu Leu Leu Lys Ala Gln Ile Ala His Phe Phe Glu His Tyr Lys Asp 
    130                 135                 140                 


Leu Glu Lys Gly Lys Trp Val Lys Val Glu Gly Trp Glu Asn Ala Glu 
145                 150                 155                 160 


Ala Ala Lys Ala Glu Ile Val Ala Ser Phe Glu Arg Ala Lys Asn Lys 
                165                 170                 175     


Cys 
    


<210>  7
<211>  177
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Mutated Escherichia coli PPase_ C54A,C88A,177C

<400>  7

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


Tyr Val Val Ile Glu Ile Pro Ala Asn Ala Asp Pro Ile Lys Tyr Glu 
            20                  25                  30          


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


Ala Met Phe Tyr Pro Ala Asn Tyr Gly Tyr Ile Asn His Thr Leu Ser 
    50                  55                  60                  


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


Gln Pro Gly Ser Val Ile Arg Ala Arg Pro Val Gly Val Leu Lys Met 
                85                  90                  95      


Thr Asp Glu Ala Gly Glu Asp Ala Lys Leu Val Ala Val Pro His Ser 
            100                 105                 110         


Lys Leu Ser Lys Glu Tyr Asp His Ile Lys Asp Val Asn Asp Leu Pro 
        115                 120                 125             


Glu Leu Leu Lys Ala Gln Ile Ala His Phe Phe Glu His Tyr Lys Asp 
    130                 135                 140                 


Leu Glu Lys Gly Lys Trp Val Lys Val Glu Gly Trp Glu Asn Ala Glu 
145                 150                 155                 160 


Ala Ala Lys Ala Glu Ile Val Ala Ser Phe Glu Arg Ala Lys Asn Lys 
                165                 170                 175     


Cys 
    


<210>  8
<211>  177
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Mutated Escherichia coli PPase_ C54A,C88S,177C

<400>  8

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


Tyr Val Val Ile Glu Ile Pro Ala Asn Ala Asp Pro Ile Lys Tyr Glu 
            20                  25                  30          


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


Ala Met Phe Tyr Pro Ala Asn Tyr Gly Tyr Ile Asn His Thr Leu Ser 
    50                  55                  60                  


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


Gln Pro Gly Ser Val Ile Arg Ser Arg Pro Val Gly Val Leu Lys Met 
                85                  90                  95      


Thr Asp Glu Ala Gly Glu Asp Ala Lys Leu Val Ala Val Pro His Ser 
            100                 105                 110         


Lys Leu Ser Lys Glu Tyr Asp His Ile Lys Asp Val Asn Asp Leu Pro 
        115                 120                 125             


Glu Leu Leu Lys Ala Gln Ile Ala His Phe Phe Glu His Tyr Lys Asp 
    130                 135                 140                 


Leu Glu Lys Gly Lys Trp Val Lys Val Glu Gly Trp Glu Asn Ala Glu 
145                 150                 155                 160 


Ala Ala Lys Ala Glu Ile Val Ala Ser Phe Glu Arg Ala Lys Asn Lys 
                165                 170                 175     


Cys 
    


<210>  9
<211>  182
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Mutated Escherichia coli PPase_ C54A,C88A,177GGGGGC

<400>  9

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


Tyr Val Val Ile Glu Ile Pro Ala Asn Ala Asp Pro Ile Lys Tyr Glu 
            20                  25                  30          


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


Ala Met Phe Tyr Pro Ala Asn Tyr Gly Tyr Ile Asn His Thr Leu Ser 
    50                  55                  60                  


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


Gln Pro Gly Ser Val Ile Arg Ala Arg Pro Val Gly Val Leu Lys Met 
                85                  90                  95      


Thr Asp Glu Ala Gly Glu Asp Ala Lys Leu Val Ala Val Pro His Ser 
            100                 105                 110         


Lys Leu Ser Lys Glu Tyr Asp His Ile Lys Asp Val Asn Asp Leu Pro 
        115                 120                 125             


Glu Leu Leu Lys Ala Gln Ile Ala His Phe Phe Glu His Tyr Lys Asp 
    130                 135                 140                 


Leu Glu Lys Gly Lys Trp Val Lys Val Glu Gly Trp Glu Asn Ala Glu 
145                 150                 155                 160 


Ala Ala Lys Ala Glu Ile Val Ala Ser Phe Glu Arg Ala Lys Asn Lys 
                165                 170                 175     


Gly Gly Gly Gly Gly Cys 
            180         


<210>  10
<211>  175
<212>  PRT
<213>  Thermus aquaticus

<400>  10

Met Ala Asn Leu Lys Ser Leu Pro Val Gly Lys Asn Ala Pro Gln Val 
1               5                   10                  15      


Val His Met Val Ile Glu Val Pro Arg Gly Ser Gly Asn Lys Tyr Glu 
            20                  25                  30          


Tyr Asp Pro Glu Leu Gly Val Val Lys Leu Asp Arg Val Leu Pro Gly 
        35                  40                  45              


Ala Gln Phe Tyr Pro Gly Asp Tyr Gly Phe Ile Pro Ser Thr Leu Ala 
    50                  55                  60                  


Glu Asp Gly Asp Pro Leu Asp Gly Leu Ile Leu Ser Thr Tyr Pro Leu 
65                  70                  75                  80  


Leu Pro Gly Val Val Val Glu Val Arg Pro Val Gly Leu Leu Leu Met 
                85                  90                  95      


Glu Asp Glu Lys Gly Gly Asp Ala Lys Ile Leu Gly Val Val Ala Glu 
            100                 105                 110         


Asp Gln Arg Leu Asp His Ile Gln Asp Ile Gly Asp Val Pro Glu Gly 
        115                 120                 125             


Val Lys Gln Glu Ile Gln His Phe Phe Glu Thr Tyr Lys Ala Leu Glu 
    130                 135                 140                 


Ala Lys Lys Gly Lys Trp Val Arg Val Thr Gly Trp Arg Asp Arg Gln 
145                 150                 155                 160 


Ala Ala Leu Glu Glu Ile Gln Ala Ala Ile Ala Arg Tyr Gly Arg 
                165                 170                 175 


<210>  11
<211>  185
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Mutated inorganic pyrophosphatase [Thermus 
       aquaticus]176GGGGSGGGGC

<400>  11

Met Ala Asn Leu Lys Ser Leu Pro Val Gly Lys Asn Ala Pro Gln Val 
1               5                   10                  15      


Val His Met Val Ile Glu Val Pro Arg Gly Ser Gly Asn Lys Tyr Glu 
            20                  25                  30          


Tyr Asp Pro Glu Leu Gly Val Val Lys Leu Asp Arg Val Leu Pro Gly 
        35                  40                  45              


Ala Gln Phe Tyr Pro Gly Asp Tyr Gly Phe Ile Pro Ser Thr Leu Ala 
    50                  55                  60                  


Glu Asp Gly Asp Pro Leu Asp Gly Leu Ile Leu Ser Thr Tyr Pro Leu 
65                  70                  75                  80  


Leu Pro Gly Val Val Val Glu Val Arg Pro Val Gly Leu Leu Leu Met 
                85                  90                  95      


Glu Asp Glu Lys Gly Gly Asp Ala Lys Ile Leu Gly Val Val Ala Glu 
            100                 105                 110         


Asp Gln Arg Leu Asp His Ile Gln Asp Ile Gly Asp Val Pro Glu Gly 
        115                 120                 125             


Val Lys Gln Glu Ile Gln His Phe Phe Glu Thr Tyr Lys Ala Leu Glu 
    130                 135                 140                 


Ala Lys Lys Gly Lys Trp Val Arg Val Thr Gly Trp Arg Asp Arg Gln 
145                 150                 155                 160 


Ala Ala Leu Glu Glu Ile Gln Ala Ala Ile Ala Arg Tyr Gly Arg Gly 
                165                 170                 175     


Gly Gly Gly Ser Gly Gly Gly Gly Cys 
            180                 185 


<210>  12
<211>  181
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Mutated inorganic pyrophosphatase [Thermus aquaticus]176GGGGGC

<400>  12

Met Ala Asn Leu Lys Ser Leu Pro Val Gly Lys Asn Ala Pro Gln Val 
1               5                   10                  15      


Val His Met Val Ile Glu Val Pro Arg Gly Ser Gly Asn Lys Tyr Glu 
            20                  25                  30          


Tyr Asp Pro Glu Leu Gly Val Val Lys Leu Asp Arg Val Leu Pro Gly 
        35                  40                  45              


Ala Gln Phe Tyr Pro Gly Asp Tyr Gly Phe Ile Pro Ser Thr Leu Ala 
    50                  55                  60                  


Glu Asp Gly Asp Pro Leu Asp Gly Leu Ile Leu Ser Thr Tyr Pro Leu 
65                  70                  75                  80  


Leu Pro Gly Val Val Val Glu Val Arg Pro Val Gly Leu Leu Leu Met 
                85                  90                  95      


Glu Asp Glu Lys Gly Gly Asp Ala Lys Ile Leu Gly Val Val Ala Glu 
            100                 105                 110         


Asp Gln Arg Leu Asp His Ile Gln Asp Ile Gly Asp Val Pro Glu Gly 
        115                 120                 125             


Val Lys Gln Glu Ile Gln His Phe Phe Glu Thr Tyr Lys Ala Leu Glu 
    130                 135                 140                 


Ala Lys Lys Gly Lys Trp Val Arg Val Thr Gly Trp Arg Asp Arg Gln 
145                 150                 155                 160 


Ala Ala Leu Glu Glu Ile Gln Ala Ala Ile Ala Arg Tyr Gly Arg Gly 
                165                 170                 175     


Gly Gly Gly Gly Cys 
            180     


<210>  13
<211>  176
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Mutated inorganic pyrophosphatase [Thermus aquaticus]176C

<400>  13

Met Ala Asn Leu Lys Ser Leu Pro Val Gly Lys Asn Ala Pro Gln Val 
1               5                   10                  15      


Val His Met Val Ile Glu Val Pro Arg Gly Ser Gly Asn Lys Tyr Glu 
            20                  25                  30          


Tyr Asp Pro Glu Leu Gly Val Val Lys Leu Asp Arg Val Leu Pro Gly 
        35                  40                  45              


Ala Gln Phe Tyr Pro Gly Asp Tyr Gly Phe Ile Pro Ser Thr Leu Ala 
    50                  55                  60                  


Glu Asp Gly Asp Pro Leu Asp Gly Leu Ile Leu Ser Thr Tyr Pro Leu 
65                  70                  75                  80  


Leu Pro Gly Val Val Val Glu Val Arg Pro Val Gly Leu Leu Leu Met 
                85                  90                  95      


Glu Asp Glu Lys Gly Gly Asp Ala Lys Ile Leu Gly Val Val Ala Glu 
            100                 105                 110         


Asp Gln Arg Leu Asp His Ile Gln Asp Ile Gly Asp Val Pro Glu Gly 
        115                 120                 125             


Val Lys Gln Glu Ile Gln His Phe Phe Glu Thr Tyr Lys Ala Leu Glu 
    130                 135                 140                 


Ala Lys Lys Gly Lys Trp Val Arg Val Thr Gly Trp Arg Asp Arg Gln 
145                 150                 155                 160 


Ala Ala Leu Glu Glu Ile Gln Ala Ala Ile Ala Arg Tyr Gly Arg Cys 
                165                 170                 175     


<210>  14
<211>  191
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Mutated inorganic pyrophosphatase [Thermus aquaticus]176(GGGGS)3C

<400>  14

Met Ala Asn Leu Lys Ser Leu Pro Val Gly Lys Asn Ala Pro Gln Val 
1               5                   10                  15      


Val His Met Val Ile Glu Val Pro Arg Gly Ser Gly Asn Lys Tyr Glu 
            20                  25                  30          


Tyr Asp Pro Glu Leu Gly Val Val Lys Leu Asp Arg Val Leu Pro Gly 
        35                  40                  45              


Ala Gln Phe Tyr Pro Gly Asp Tyr Gly Phe Ile Pro Ser Thr Leu Ala 
    50                  55                  60                  


Glu Asp Gly Asp Pro Leu Asp Gly Leu Ile Leu Ser Thr Tyr Pro Leu 
65                  70                  75                  80  


Leu Pro Gly Val Val Val Glu Val Arg Pro Val Gly Leu Leu Leu Met 
                85                  90                  95      


Glu Asp Glu Lys Gly Gly Asp Ala Lys Ile Leu Gly Val Val Ala Glu 
            100                 105                 110         


Asp Gln Arg Leu Asp His Ile Gln Asp Ile Gly Asp Val Pro Glu Gly 
        115                 120                 125             


Val Lys Gln Glu Ile Gln His Phe Phe Glu Thr Tyr Lys Ala Leu Glu 
    130                 135                 140                 


Ala Lys Lys Gly Lys Trp Val Arg Val Thr Gly Trp Arg Asp Arg Gln 
145                 150                 155                 160 


Ala Ala Leu Glu Glu Ile Gln Ala Ala Ile Ala Arg Tyr Gly Arg Gly 
                165                 170                 175     


Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Cys 
            180                 185                 190     


<210>  15
<211>  175
<212>  PRT
<213>  Thermus thermophilus

<400>  15

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


Val His Met Val Ile Glu Val Pro Arg Gly Ser Gly Asn Lys Tyr Glu 
            20                  25                  30          


Tyr Asp Pro Asp Leu Gly Ala Ile Lys Leu Asp Arg Val Leu Pro Gly 
        35                  40                  45              


Ala Gln Phe Tyr Pro Gly Asp Tyr Gly Phe Ile Pro Ser Thr Leu Ala 
    50                  55                  60                  


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


Leu Pro Gly Val Val Val Glu Val Arg Val Val Gly Leu Leu Leu Met 
                85                  90                  95      


Glu Asp Glu Lys Gly Gly Asp Ala Lys Val Ile Gly Val Val Ala Glu 
            100                 105                 110         


Asp Gln Arg Leu Asp His Ile Gln Asp Ile Gly Asp Val Pro Glu Gly 
        115                 120                 125             


Val Lys Gln Glu Ile Gln His Phe Phe Glu Thr Tyr Lys Ala Leu Glu 
    130                 135                 140                 


Ala Lys Lys Gly Lys Trp Val Lys Val Thr Gly Trp Arg Asp Arg Lys 
145                 150                 155                 160 


Ala Ala Leu Glu Glu Val Arg Ala Cys Ile Ala Arg Tyr Lys Gly 
                165                 170                 175 


<210>  16
<211>  176
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Mutated inorganic pyrophosphatase [Thermus 
       thermophilus]_C169A,176C

<400>  16

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


Val His Met Val Ile Glu Val Pro Arg Gly Ser Gly Asn Lys Tyr Glu 
            20                  25                  30          


Tyr Asp Pro Asp Leu Gly Ala Ile Lys Leu Asp Arg Val Leu Pro Gly 
        35                  40                  45              


Ala Gln Phe Tyr Pro Gly Asp Tyr Gly Phe Ile Pro Ser Thr Leu Ala 
    50                  55                  60                  


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


Leu Pro Gly Val Val Val Glu Val Arg Val Val Gly Leu Leu Leu Met 
                85                  90                  95      


Glu Asp Glu Lys Gly Gly Asp Ala Lys Val Ile Gly Val Val Ala Glu 
            100                 105                 110         


Asp Gln Arg Leu Asp His Ile Gln Asp Ile Gly Asp Val Pro Glu Gly 
        115                 120                 125             


Val Lys Gln Glu Ile Gln His Phe Phe Glu Thr Tyr Lys Ala Leu Glu 
    130                 135                 140                 


Ala Lys Lys Gly Lys Trp Val Lys Val Thr Gly Trp Arg Asp Arg Lys 
145                 150                 155                 160 


Ala Ala Leu Glu Glu Val Arg Ala Ala Ile Ala Arg Tyr Lys Gly Cys 
                165                 170                 175     


<210>  17
<211>  176
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Mutated inorganic pyrophosphatase [Thermus 
       thermophilus]_C169S,176C

<400>  17

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


Val His Met Val Ile Glu Val Pro Arg Gly Ser Gly Asn Lys Tyr Glu 
            20                  25                  30          


Tyr Asp Pro Asp Leu Gly Ala Ile Lys Leu Asp Arg Val Leu Pro Gly 
        35                  40                  45              


Ala Gln Phe Tyr Pro Gly Asp Tyr Gly Phe Ile Pro Ser Thr Leu Ala 
    50                  55                  60                  


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


Leu Pro Gly Val Val Val Glu Val Arg Val Val Gly Leu Leu Leu Met 
                85                  90                  95      


Glu Asp Glu Lys Gly Gly Asp Ala Lys Val Ile Gly Val Val Ala Glu 
            100                 105                 110         


Asp Gln Arg Leu Asp His Ile Gln Asp Ile Gly Asp Val Pro Glu Gly 
        115                 120                 125             


Val Lys Gln Glu Ile Gln His Phe Phe Glu Thr Tyr Lys Ala Leu Glu 
    130                 135                 140                 


Ala Lys Lys Gly Lys Trp Val Lys Val Thr Gly Trp Arg Asp Arg Lys 
145                 150                 155                 160 


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


<210>  18
<211>  191
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Mutated inorganic pyrophosphatase [Thermus 
       thermophilus]_C169A,176(GGGGS)3C

<400>  18

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


Val His Met Val Ile Glu Val Pro Arg Gly Ser Gly Asn Lys Tyr Glu 
            20                  25                  30          


Tyr Asp Pro Asp Leu Gly Ala Ile Lys Leu Asp Arg Val Leu Pro Gly 
        35                  40                  45              


Ala Gln Phe Tyr Pro Gly Asp Tyr Gly Phe Ile Pro Ser Thr Leu Ala 
    50                  55                  60                  


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


Leu Pro Gly Val Val Val Glu Val Arg Val Val Gly Leu Leu Leu Met 
                85                  90                  95      


Glu Asp Glu Lys Gly Gly Asp Ala Lys Val Ile Gly Val Val Ala Glu 
            100                 105                 110         


Asp Gln Arg Leu Asp His Ile Gln Asp Ile Gly Asp Val Pro Glu Gly 
        115                 120                 125             


Val Lys Gln Glu Ile Gln His Phe Phe Glu Thr Tyr Lys Ala Leu Glu 
    130                 135                 140                 


Ala Lys Lys Gly Lys Trp Val Lys Val Thr Gly Trp Arg Asp Arg Lys 
145                 150                 155                 160 


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


Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Cys 
            180                 185                 190     


<210>  19
<211>  191
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Mutated inorganic pyrophosphatase [Thermus thermophilus]_C169S, 
       176(GGGGS)3C

<400>  19

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


Val His Met Val Ile Glu Val Pro Arg Gly Ser Gly Asn Lys Tyr Glu 
            20                  25                  30          


Tyr Asp Pro Asp Leu Gly Ala Ile Lys Leu Asp Arg Val Leu Pro Gly 
        35                  40                  45              


Ala Gln Phe Tyr Pro Gly Asp Tyr Gly Phe Ile Pro Ser Thr Leu Ala 
    50                  55                  60                  


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


Leu Pro Gly Val Val Val Glu Val Arg Val Val Gly Leu Leu Leu Met 
                85                  90                  95      


Glu Asp Glu Lys Gly Gly Asp Ala Lys Val Ile Gly Val Val Ala Glu 
            100                 105                 110         


Asp Gln Arg Leu Asp His Ile Gln Asp Ile Gly Asp Val Pro Glu Gly 
        115                 120                 125             


Val Lys Gln Glu Ile Gln His Phe Phe Glu Thr Tyr Lys Ala Leu Glu 
    130                 135                 140                 


Ala Lys Lys Gly Lys Trp Val Lys Val Thr Gly Trp Arg Asp Arg Lys 
145                 150                 155                 160 


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


Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Cys 
            180                 185                 190     


<210>  20
<211>  181
<212>  PRT
<213>  Artificial Sequnece

<220>
<223>  Mutated inorganic pyrophosphatase [Thermus 
       thermophilus]_C169A,176GGGGGC

<400>  20

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


Val His Met Val Ile Glu Val Pro Arg Gly Ser Gly Asn Lys Tyr Glu 
            20                  25                  30          


Tyr Asp Pro Asp Leu Gly Ala Ile Lys Leu Asp Arg Val Leu Pro Gly 
        35                  40                  45              


Ala Gln Phe Tyr Pro Gly Asp Tyr Gly Phe Ile Pro Ser Thr Leu Ala 
    50                  55                  60                  


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


Leu Pro Gly Val Val Val Glu Val Arg Val Val Gly Leu Leu Leu Met 
                85                  90                  95      


Glu Asp Glu Lys Gly Gly Asp Ala Lys Val Ile Gly Val Val Ala Glu 
            100                 105                 110         


Asp Gln Arg Leu Asp His Ile Gln Asp Ile Gly Asp Val Pro Glu Gly 
        115                 120                 125             


Val Lys Gln Glu Ile Gln His Phe Phe Glu Thr Tyr Lys Ala Leu Glu 
    130                 135                 140                 


Ala Lys Lys Gly Lys Trp Val Lys Val Thr Gly Trp Arg Asp Arg Lys 
145                 150                 155                 160 


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


Gly Gly Gly Gly Cys 
            180     


<210>  21
<211>  181
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Inorganic pyrophosphatase [Thermus thermophilus]_C169A, 176GGGGGC

<400>  21

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


Val His Met Val Ile Glu Val Pro Arg Gly Ser Gly Asn Lys Tyr Glu 
            20                  25                  30          


Tyr Asp Pro Asp Leu Gly Ala Ile Lys Leu Asp Arg Val Leu Pro Gly 
        35                  40                  45              


Ala Gln Phe Tyr Pro Gly Asp Tyr Gly Phe Ile Pro Ser Thr Leu Ala 
    50                  55                  60                  


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


Leu Pro Gly Val Val Val Glu Val Arg Val Val Gly Leu Leu Leu Met 
                85                  90                  95      


Glu Asp Glu Lys Gly Gly Asp Ala Lys Val Ile Gly Val Val Ala Glu 
            100                 105                 110         


Asp Gln Arg Leu Asp His Ile Gln Asp Ile Gly Asp Val Pro Glu Gly 
        115                 120                 125             


Val Lys Gln Glu Ile Gln His Phe Phe Glu Thr Tyr Lys Ala Leu Glu 
    130                 135                 140                 


Ala Lys Lys Gly Lys Trp Val Lys Val Thr Gly Trp Arg Asp Arg Lys 
145                 150                 155                 160 


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


Gly Gly Gly Gly Cys 
            180     


<210>  22
<211>  1653
<212>  RNA
<213>  Artificial sequence

<220>
<223>  PPLuc modified coding sequence

<400>  22
auggaggacg ccaagaacau caagaagggc ccggcgcccu ucuacccgcu ggaggacggg       60

accgccggcg agcagcucca caaggccaug aagcgguacg cccuggugcc gggcacgauc      120

gccuucaccg acgcccacau cgaggucgac aucaccuacg cggaguacuu cgagaugagc      180

gugcgccugg ccgaggccau gaagcgguac ggccugaaca ccaaccaccg gaucguggug      240

ugcucggaga acagccugca guucuucaug ccggugcugg gcgcccucuu caucggcgug      300

gccgucgccc cggcgaacga caucuacaac gagcgggagc ugcugaacag cauggggauc      360

agccagccga ccgugguguu cgugagcaag aagggccugc agaagauccu gaacgugcag      420

aagaagcugc ccaucaucca gaagaucauc aucauggaca gcaagaccga cuaccagggc      480

uuccagucga uguacacguu cgugaccagc caccucccgc cgggcuucaa cgaguacgac      540

uucgucccgg agagcuucga ccgggacaag accaucgccc ugaucaugaa cagcagcggc      600

agcaccggcc ugccgaaggg gguggcccug ccgcaccgga ccgccugcgu gcgcuucucg      660

cacgcccggg accccaucuu cggcaaccag aucaucccgg acaccgccau ccugagcgug      720

gugccguucc accacggcuu cggcauguuc acgacccugg gcuaccucau cugcggcuuc      780

cggguggucc ugauguaccg guucgaggag gagcuguucc ugcggagccu gcaggacuac      840

aagauccaga gcgcgcugcu cgugccgacc cuguucagcu ucuucgccaa gagcacccug      900

aucgacaagu acgaccuguc gaaccugcac gagaucgcca gcgggggcgc cccgcugagc      960

aaggaggugg gcgaggccgu ggccaagcgg uuccaccucc cgggcauccg ccagggcuac     1020

ggccugaccg agaccacgag cgcgauccug aucacccccg agggggacga caagccgggc     1080

gccgugggca aggugguccc guucuucgag gccaaggugg uggaccugga caccggcaag     1140

acccugggcg ugaaccagcg gggcgagcug ugcgugcggg ggccgaugau caugagcggc     1200

uacgugaaca acccggaggc caccaacgcc cucaucgaca aggacggcug gcugcacagc     1260

ggcgacaucg ccuacuggga cgaggacgag cacuucuuca ucgucgaccg gcugaagucg     1320

cugaucaagu acaagggcua ccagguggcg ccggccgagc uggagagcau ccugcuccag     1380

caccccaaca ucuucgacgc cggcguggcc gggcugccgg acgacgacgc cggcgagcug     1440

ccggccgcgg ugguggugcu ggagcacggc aagaccauga cggagaagga gaucgucgac     1500

uacguggcca gccaggugac caccgccaag aagcugcggg gcggcguggu guucguggac     1560

gaggucccga agggccugac cgggaagcuc gacgcccgga agauccgcga gauccugauc     1620

aaggccaaga agggcggcaa gaucgccgug uga                                  1653


