                         SEQUENCE LISTING

<110>  Nuclera Nucleics Ltd
 
<120>  Monitoring of in vitro protein synthesis

<130>  P33326WO1

<140>  PCT/GB2021/052140
<141>  2021-08-18

<150>  GB2013063.9
<151>  2020-08-21

<160>  12    

<170>  PatentIn version 3.5

<210>  1
<211>  403
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  GFP11_TdT fusion protein sequence

<400>  1

Met Lys Arg Asp His Met Val Leu His Glu Tyr Val Asn Ala Ala Gly 
1               5                   10                  15      


Ile Thr Gly Ser Gly Gly Ser Gly Gly Lys Phe Met His His His His 
            20                  25                  30          


His His Met Glu Asn Leu Tyr Phe Gln Gly Lys Ile Ser Gln Tyr Ala 
        35                  40                  45              


Cys Gln Arg Lys Thr Thr Leu Asn Asn Tyr Asn His Ile Phe Thr Asp 
    50                  55                  60                  


Ala Phe Glu Ile Leu Ala Glu Asn Ser Glu Phe Lys Glu Asn Glu Val 
65                  70                  75                  80  


Ser Tyr Val Thr Phe Met Arg Ala Ala Ser Val Leu Lys Ser Leu Pro 
                85                  90                  95      


Phe Thr Ile Ile Ser Met Lys Asp Thr Glu Gly Ile Pro Cys Leu Gly 
            100                 105                 110         


Asp Lys Val Lys Cys Ile Ile Glu Glu Ile Ile Glu Asp Gly Glu Ser 
        115                 120                 125             


Ser Glu Val Lys Ala Val Leu Asn Asp Glu Arg Tyr Gln Ser Phe Lys 
    130                 135                 140                 


Leu Phe Thr Ser Val Phe Gly Val Gly Leu Lys Thr Ser Glu Lys Trp 
145                 150                 155                 160 


Phe Arg Met Gly Phe Arg Ser Leu Ser Lys Ile Met Ser Asp Lys Thr 
                165                 170                 175     


Leu Lys Phe Thr Lys Met Gln Lys Ala Gly Phe Leu Tyr Tyr Glu Asp 
            180                 185                 190         


Leu Val Ser Cys Val Thr Arg Ala Glu Ala Glu Ala Val Gly Val Leu 
        195                 200                 205             


Val Lys Glu Ala Val Trp Ala Phe Leu Pro Asp Ala Phe Val Thr Met 
    210                 215                 220                 


Thr Gly Gly Phe Arg Arg Gly Lys Lys Ile Gly His Asp Val Asp Phe 
225                 230                 235                 240 


Leu Ile Thr Ser Pro Gly Ser Ala Glu Asp Glu Glu Gln Leu Leu Pro 
                245                 250                 255     


Lys Val Ile Asn Leu Trp Glu Lys Lys Gly Leu Leu Leu Tyr Tyr Asp 
            260                 265                 270         


Leu Val Glu Ser Thr Phe Glu Lys Phe Lys Leu Pro Ser Arg Gln Val 
        275                 280                 285             


Asp Thr Leu Asp His Phe Gln Lys Cys Phe Leu Ile Leu Lys Leu His 
    290                 295                 300                 


His Gln Arg Val Asp Ser Ser Lys Ser Asn Gln Gln Glu Gly Lys Thr 
305                 310                 315                 320 


Trp Lys Ala Ile Arg Val Asp Leu Val Met Cys Pro Tyr Glu Asn Arg 
                325                 330                 335     


Ala Phe Ala Leu Leu Gly Trp Thr Gly Ser Arg Gln Phe Glu Arg Asp 
            340                 345                 350         


Ile Arg Arg Tyr Ala Thr His Glu Arg Lys Met Met Leu Asp Asn His 
        355                 360                 365             


Ala Leu Tyr Asp Lys Thr Lys Arg Val Phe Leu Lys Ala Glu Ser Glu 
    370                 375                 380                 


Glu Glu Ile Phe Ala His Leu Gly Leu Asp Tyr Ile Glu Pro Trp Glu 
385                 390                 395                 400 


Arg Asn Ala 
            


<210>  2
<211>  214
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  sfGFG GFP1-10 polypeptide AA sequence

<400>  2

Met Ser Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro Ile Leu Val 
1               5                   10                  15      


Glu Leu Asp Gly Asp Val Asn Gly His Lys Phe Ser Val Arg Gly Glu 
            20                  25                  30          


Gly Glu Gly Asp Ala Thr Asn Gly Lys Leu Thr Leu Lys Phe Ile Cys 
        35                  40                  45              


Thr Thr Gly Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr Leu 
    50                  55                  60                  


Thr Tyr Gly Val Gln Cys Phe Ser Arg Tyr Pro Asp His Met Lys Arg 
65                  70                  75                  80  


His Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu Arg 
                85                  90                  95      


Thr Ile Ser Phe Lys Asp Asp Gly Thr Tyr Lys Thr Arg Ala Glu Val 
            100                 105                 110         


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


Asp Phe Lys Glu Asp Gly Asn Ile Leu Gly His Lys Leu Glu Tyr Asn 
    130                 135                 140                 


Phe Asn Ser His Asn Val Tyr Ile Thr Ala Asp Lys Gln Lys Asn Gly 
145                 150                 155                 160 


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


Gln Leu Ala Asp His Tyr Gln Gln Asn Thr Pro Ile Gly Asp Gly Pro 
            180                 185                 190         


Val Leu Leu Pro Asp Asn His Tyr Leu Ser Thr Gln Ser Val Leu Ser 
        195                 200                 205             


Lys Asp Pro Asn Glu Lys 
    210                 


<210>  3
<211>  214
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Mutated sfGFP GFP1-10 polynucleotide AA sequence

<400>  3

Met Ser Lys Gly Glu Glu Leu Phe Thr Gly Val Val Pro Ile Leu Val 
1               5                   10                  15      


Glu Leu Asp Gly Asp Val Asn Gly His Lys Phe Ser Val Arg Gly Glu 
            20                  25                  30          


Gly Glu Gly Asp Ala Thr Ile Gly Lys Leu Thr Leu Lys Phe Ile Cys 
        35                  40                  45              


Thr Thr Gly Lys Leu Pro Val Pro Trp Pro Thr Leu Val Thr Thr Leu 
    50                  55                  60                  


Thr Tyr Gly Val Gln Cys Phe Ser Arg Tyr Pro Asp His Met Lys Arg 
65                  70                  75                  80  


His Asp Phe Phe Lys Ser Ala Met Pro Glu Gly Tyr Val Gln Glu Arg 
                85                  90                  95      


Thr Ile Ser Phe Lys Asp Asp Gly Lys Tyr Lys Thr Arg Ala Val Val 
            100                 105                 110         


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


Asp Phe Lys Glu Asp Gly Asn Ile Leu Gly His Lys Leu Glu Tyr Asn 
    130                 135                 140                 


Phe Asn Ser His Asn Val Tyr Ile Thr Ala Asp Lys Gln Lys Asn Gly 
145                 150                 155                 160 


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


Gln Leu Ala Asp His Tyr Gln Gln Asn Thr Pro Ile Gly Asp Gly Pro 
            180                 185                 190         


Val Leu Leu Pro Asp Asn His Tyr Leu Ser Thr Gln Thr Val Leu Ser 
        195                 200                 205             


Lys Asp Pro Asn Glu Lys 
    210                 


<210>  4
<211>  19
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  GFP11 1.0 sequence

<400>  4

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


Thr Gly Thr 
            


<210>  5
<211>  19
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  GFP11 2.0 sequence

<400>  5

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


Thr Gly Thr 
            


<210>  6
<211>  17
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  GFP11 3.0 sequence

<400>  6

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


Thr 
    


<210>  7
<211>  16
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  GFP11 4.0 sequence

<400>  7

Arg Asp His Met Val Leu His Glu Tyr Val Asn Ala Ala Gly Ile Thr 
1               5                   10                  15      


<210>  8
<211>  208
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  sfCherry1-10 polypeptide AA sequence

<400>  8

Met Glu Glu Asp Asn Met Ala Ile Ile Lys Glu Phe Met Arg Phe Lys 
1               5                   10                  15      


Val His Met Glu Gly Ser Val Asn Gly His Glu Phe Glu Ile Glu Gly 
            20                  25                  30          


Glu Gly Glu Gly His Pro Tyr Glu Gly Thr Gln Thr Ala Lys Leu Lys 
        35                  40                  45              


Val Thr Lys Gly Gly Pro Leu Pro Phe Ala Trp Asp Ile Leu Ser Pro 
    50                  55                  60                  


Gln Phe Met Tyr Gly Ser Lys Ala Tyr Val Lys His Pro Ala Asp Ile 
65                  70                  75                  80  


Pro Asp Tyr Leu Lys Leu Ser Phe Pro Glu Gly Phe Thr Trp Glu Arg 
                85                  90                  95      


Val Met Asn Phe Glu Asp Gly Gly Val Val Thr Val Thr Gln Asp Ser 
            100                 105                 110         


Ser Leu Gln Asp Gly Glu Phe Ile Tyr Lys Val Lys Leu Leu Gly Thr 
        115                 120                 125             


Asn Phe Pro Ser Asp Gly Pro Val Met Gln Lys Lys Thr Met Gly Trp 
    130                 135                 140                 


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


Glu Ile Asn Gln Arg Leu Lys Leu Lys Asp Gly Gly His Tyr Asp Ala 
                165                 170                 175     


Glu Val Lys Thr Thr Tyr Lys Ala Lys Lys Pro Val Gln Leu Pro Gly 
            180                 185                 190         


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


<210>  9
<211>  18
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  sfCherry11 sequence

<400>  9

Tyr Thr Ile Val Glu Gln Tyr Glu Arg Ala Glu Gly Arg His Ser Thr 
1               5                   10                  15      


Gly Gly 
        


<210>  10
<211>  114
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  NFAST sequence

<400>  10

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


Lys Met Asp Asp Gly Gln Leu Asp Gly Leu Ala Phe Gly Ala Ile Gln 
            20                  25                  30          


Leu Asp Gly Asp Gly Asn Ile Leu Gln Tyr Asn Ala Ala Glu Gly Asp 
        35                  40                  45              


Ile Thr Gly Arg Asp Pro Lys Gln Val Ile Gly Lys Asn Phe Phe Lys 
    50                  55                  60                  


Asp Val Ala Pro Gly Thr Asp Ser Pro Glu Phe Tyr Gly Lys Phe Lys 
65                  70                  75                  80  


Glu Gly Val Ala Ser Gly Asn Leu Asn Thr Met Phe Glu Trp Met Ile 
                85                  90                  95      


Pro Thr Ser Arg Gly Pro Thr Lys Val Lys Val His Met Lys Lys Ala 
            100                 105                 110         


Leu Ser 
        


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

<220>
<223>  CFAST11 sequence

<400>  11

Gly Asp Ser Tyr Trp Val Phe Val Lys Arg Val 
1               5                   10      


<210>  12
<211>  10
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  CFAST10 sequence

<400>  12

Gly Asp Ser Tyr Trp Val Phe Val Lys Arg 
1               5                   10  


