                         SEQUENCE LISTING



<110>  Brigham Young University

 

<120>  METHOD FOR DETECTION OF VIRAL INFECTIONS USING SPLIT ENZYMES IN 

       AN OXIDATIVE ENVIRONMENT



<130>  82570-344340



<150>  63/121,784

<151>  2020-12-04



<150>  63/260,247

<151>  2021-08-13



<160>  6     



<170>  PatentIn version 3.5



<210>  1

<211>  481

<212>  PRT

<213>  Artificial Sequence



<220>

<223>  Split HRP 1 with COVID19 scfv



<400>  1



Met His His His His His His Met Glu Val Gln Leu Val Glu Ser Gly 

1               5                   10                  15      





Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala 

            20                  25                  30          





Ser Gly Phe Ile Val Ser Ser Asn Tyr Met Ser Trp Val Arg Gln Ala 

        35                  40                  45              





Pro Gly Lys Gly Leu Glu Trp Val Ser Val Ile Tyr Ser Gly Gly Ser 

    50                  55                  60                  





Thr Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg His 

65                  70                  75                  80  





Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu 

                85                  90                  95      





Asp Thr Ala Val Tyr Tyr Cys Ala Arg Glu Ala Tyr Gly Met Asp Val 

            100                 105                 110         





Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser 

        115                 120                 125             





Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Val Met Thr Gln 

    130                 135                 140                 





Ser Pro Ser Phe Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr 

145                 150                 155                 160 





Cys Arg Ala Ser Gln Gly Ile Ser Ser Tyr Leu Ala Trp Tyr Gln Gln 

                165                 170                 175     





Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr Ala Ala Ser Thr Leu 

            180                 185                 190         





Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu 

        195                 200                 205             





Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr 

    210                 215                 220                 





Tyr Cys Gln Gln Leu Asn Ser Tyr Pro Pro Tyr Thr Phe Gly Gln Gly 

225                 230                 235                 240 





Thr Lys Leu Glu Ile Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 

                245                 250                 255     





Gly Gly Gly Gly Ser Gln Leu Thr Pro Thr Phe Tyr Asp Asn Ser Cys 

            260                 265                 270         





Pro Asn Val Ser Asn Ile Val Arg Asp Ile Ile Val Asn Glu Leu Arg 

        275                 280                 285             





Ser Asp Pro Arg Ile Ala Ala Ser Ile Leu Arg Leu His Phe His Asp 

    290                 295                 300                 





Cys Phe Val Asn Gly Cys Asp Ala Ser Ile Leu Leu Asp Asn Thr Thr 

305                 310                 315                 320 





Ser Phe Arg Thr Glu Lys Asp Ala Phe Gly Asn Ala Asn Ser Ala Arg 

                325                 330                 335     





Gly Phe Ser Val Ile Asp Arg Met Lys Ala Ala Val Glu Ser Ala Cys 

            340                 345                 350         





Pro Gly Thr Val Ser Cys Ala Asp Leu Leu Thr Ile Ala Ala Gln Gln 

        355                 360                 365             





Ser Val Thr Leu Ala Gly Gly Pro Ser Trp Arg Val Pro Leu Gly Arg 

    370                 375                 380                 





Arg Asp Ser Leu Gln Ala Phe Leu Asp Leu Ala Asn Ala Asn Leu Pro 

385                 390                 395                 400 





Ala Pro Phe Phe Thr Leu Pro Gln Leu Lys Asp Ser Phe Arg Asn Val 

                405                 410                 415     





Gly Leu Asn Arg Ser Ser Asp Leu Val Ala Leu Ser Gly Gly His Thr 

            420                 425                 430         





Phe Gly Lys Ser Gln Cys Arg Phe Ile Met Asp Arg Leu Tyr Asn Phe 

        435                 440                 445             





Ser Asn Thr Gly Leu Pro Asp Pro Thr Leu Asn Thr Thr Tyr Leu Gln 

    450                 455                 460                 





Thr Leu Arg Gly Leu Cys Pro Leu Asn Gly Gly Ser Gly Lys Asp Glu 

465                 470                 475                 480 





Leu 

    





<210>  2

<211>  379

<212>  PRT

<213>  Artificial Sequence



<220>

<223>  Split HRP 2 with COVID19 scfv



<400>  2



Met His His His His His His Met Gln Val Gln Leu Val Gln Ser Gly 

1               5                   10                  15      





Ala Glu Val Lys Lys Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala 

            20                  25                  30          





Ser Gly Tyr Thr Phe Thr Gly Tyr Tyr Met His Trp Val Arg Gln Ala 

        35                  40                  45              





Pro Gly Gln Gly Leu Glu Trp Met Gly Arg Ile Asn Pro Asn Ser Gly 

    50                  55                  60                  





Gly Thr Asn Tyr Ala Gln Lys Phe Gln Gly Arg Val Thr Met Thr Arg 

65                  70                  75                  80  





Asp Thr Ser Ile Ser Thr Ala Tyr Met Glu Leu Ser Arg Leu Arg Ser 

                85                  90                  95      





Asp Asp Thr Ala Val Tyr Tyr Cys Ala Arg Val Pro Tyr Cys Ser Ser 

            100                 105                 110         





Thr Ser Cys His Arg Asp Trp Tyr Phe Asp Leu Trp Gly Arg Gly Thr 

        115                 120                 125             





Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 

    130                 135                 140                 





Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Leu Ser Leu 

145                 150                 155                 160 





Pro Val Thr Pro Gly Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln 

                165                 170                 175     





Ser Leu Leu Asp Ser Asp Asp Gly Asn Thr Tyr Leu Asp Trp Tyr Leu 

            180                 185                 190         





Gln Lys Pro Gly Gln Ser Pro Gln Leu Leu Ile Tyr Thr Leu Ser Tyr 

        195                 200                 205             





Arg Ala Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr 

    210                 215                 220                 





Asp Phe Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val 

225                 230                 235                 240 





Tyr Tyr Cys Met Gln Arg Ile Glu Phe Pro Leu Thr Phe Gly Gly Gly 

                245                 250                 255     





Thr Lys Val Glu Ile Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 

            260                 265                 270         





Gly Gly Gly Gly Ser Asn Leu Ser Ala Leu Val Asp Phe Asp Leu Arg 

        275                 280                 285             





Thr Pro Thr Ile Phe Asp Asn Lys Tyr Tyr Val Asn Leu Glu Glu Gln 

    290                 295                 300                 





Lys Gly Leu Ile Gln Ser Asp Gln Glu Leu Phe Ser Ser Pro Asp Ala 

305                 310                 315                 320 





Thr Asp Thr Ile Pro Leu Val Arg Ser Phe Ala Asn Ser Thr Gln Thr 

                325                 330                 335     





Phe Phe Asn Ala Phe Val Glu Ala Met Asp Arg Met Gly Asn Ile Thr 

            340                 345                 350         





Pro Leu Thr Gly Thr Gln Gly Gln Ile Arg Arg Asn Cys Arg Val Val 

        355                 360                 365             





Asn Ser Asn Ser Gly Ser Gly Lys Asp Glu Leu 

    370                 375                 





<210>  3

<211>  534

<212>  PRT

<213>  Artificial Sequence



<220>

<223>  Split Apex 1 with COVID19 scfv



<400>  3



Met Gly Leu Asn Asp Ile Phe Glu Ala Gln Lys Ile Glu Trp His Glu 

1               5                   10                  15      





Gly Gly Gly Gly Ser Met Gly Lys Ser Tyr Pro Thr Val Ser Ala Asp 

            20                  25                  30          





Tyr Gln Asp Ala Val Glu Lys Ala Lys Lys Arg Leu Gly Gly Phe Ile 

        35                  40                  45              





Ala Glu Lys Arg Cys Ala Pro Leu Met Leu Arg Leu Ala Phe His Ser 

    50                  55                  60                  





Ala Gly Thr Phe Asp Lys Arg Thr Lys Thr Gly Gly Pro Phe Gly Thr 

65                  70                  75                  80  





Ile Arg Tyr Pro Ala Glu Leu Ala His Ser Ala Asn Ser Gly Leu Asp 

                85                  90                  95      





Ile Ala Val Arg Leu Leu Glu Pro Leu Lys Ala Glu Phe Pro Ile Leu 

            100                 105                 110         





Ser Tyr Ala Asp Phe Tyr Gln Leu Ala Gly Val Val Ala Val Glu Val 

        115                 120                 125             





Thr Gly Gly Pro Lys Val Pro Phe His Pro Gly Arg Glu Asp Lys Pro 

    130                 135                 140                 





Glu Leu Pro Pro Glu Gly Arg Leu Pro Asp Pro Thr Lys Gly Ser Asp 

145                 150                 155                 160 





His Leu Arg Asp Val Phe Gly Lys Ala Met Gly Leu Thr Asp Gln Asp 

                165                 170                 175     





Ile Val Ala Leu Ser Gly Gly His Thr Leu Gly Ala Ala His Lys Glu 

            180                 185                 190         





Arg Ser Gly Phe Glu Gly Pro Trp Thr Ser Asn Pro Leu Val Phe Asp 

        195                 200                 205             





Asn Ser Tyr Phe Thr Glu Leu Leu Ser Gly Glu Lys Glu Gly Gly Gly 

    210                 215                 220                 





Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Cys Ser Gly Gly 

225                 230                 235                 240 





Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 

                245                 250                 255     





Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Met Asp Ile Gln 

            260                 265                 270         





Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly Glu Pro Ala 

        275                 280                 285             





Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu Asp Ser Asp Asp Gly 

    290                 295                 300                 





Asn Thr Tyr Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser Pro Gln 

305                 310                 315                 320 





Leu Leu Ile Tyr Thr Leu Ser Tyr Arg Ala Ser Gly Val Pro Asp Arg 

                325                 330                 335     





Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile Ser Arg 

            340                 345                 350         





Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Arg Ile Glu 

        355                 360                 365             





Phe Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Ser Gly 

    370                 375                 380                 





Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gln Val 

385                 390                 395                 400 





Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala Ser Val 

                405                 410                 415     





Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Gly Tyr Tyr Met 

            420                 425                 430         





His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met Gly Arg 

        435                 440                 445             





Ile Asn Pro Asn Ser Gly Gly Thr Asn Tyr Ala Gln Lys Phe Gln Gly 

    450                 455                 460                 





Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr Met Glu 

465                 470                 475                 480 





Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys Ala Arg 

                485                 490                 495     





Val Pro Tyr Cys Ser Ser Thr Ser Cys His Arg Asp Trp Tyr Phe Asp 

            500                 505                 510         





Leu Trp Gly Arg Gly Thr Leu Val Thr Val Ser Gly Gly Gly Gly Ser 

        515                 520                 525             





His His His His His His 

    530                 





<210>  4

<211>  362

<212>  PRT

<213>  Artificial Sequence



<220>

<223>  Split Apex 2 with COVID19 scfv



<400>  4



Met His His His His His His Gly Gly Gly Gly Ser Met Glu Val Gln 

1               5                   10                  15      





Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg 

            20                  25                  30          





Leu Ser Cys Ala Ala Ser Gly Phe Ile Val Ser Ser Asn Tyr Met Ser 

        35                  40                  45              





Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Val Ile 

    50                  55                  60                  





Tyr Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe 

65                  70                  75                  80  





Thr Ile Ser Arg His Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn 

                85                  90                  95      





Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Glu Ala 

            100                 105                 110         





Tyr Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 

        115                 120                 125             





Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp 

    130                 135                 140                 





Ile Val Met Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly Asp 

145                 150                 155                 160 





Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Tyr Leu 

                165                 170                 175     





Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 

            180                 185                 190         





Ala Ala Ser Thr Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 

        195                 200                 205             





Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 

    210                 215                 220                 





Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Leu Asn Ser Tyr Pro Pro Tyr 

225                 230                 235                 240 





Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Gly Gly Gly Gly Ser 

                245                 250                 255     





Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 

            260                 265                 270         





Gly Gly Gly Ser Gly Gly Gly Gly Ser Cys Ser Gly Gly Gly Gly Ser 

        275                 280                 285             





Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala Ser Gly Leu Leu Gln 

    290                 295                 300                 





Leu Pro Ser Asp Lys Ala Leu Leu Ser Asp Pro Val Phe Arg Pro Leu 

305                 310                 315                 320 





Val Asp Lys Tyr Ala Ala Asp Glu Asp Ala Phe Phe Ala Asp Tyr Ala 

                325                 330                 335     





Glu Ala His Gln Lys Leu Ser Glu Leu Gly Phe Ala Asp Ala Leu Gln 

            340                 345                 350         





Leu Pro Pro Leu Glu Arg Leu Thr Leu Asp 

        355                 360         





<210>  5

<211>  364

<212>  PRT

<213>  Artificial Sequence



<220>

<223>  Split GFP 1 with COVID19 scfv



<400>  5



Met His His His His His His Met Glu Val Gln Leu Val Glu Ser Gly 

1               5                   10                  15      





Gly Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala 

            20                  25                  30          





Ser Gly Phe Ile Val Ser Ser Asn Tyr Met Ser Trp Val Arg Gln Ala 

        35                  40                  45              





Pro Gly Lys Gly Leu Glu Trp Val Ser Val Ile Tyr Ser Gly Gly Ser 

    50                  55                  60                  





Thr Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg His 

65                  70                  75                  80  





Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu 

                85                  90                  95      





Asp Thr Ala Val Tyr Tyr Cys Ala Arg Glu Ala Tyr Gly Met Asp Val 

            100                 105                 110         





Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Gly Gly Gly Ser 

        115                 120                 125             





Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Asp Ile Val Met Thr Gln 

    130                 135                 140                 





Ser Pro Ser Phe Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr 

145                 150                 155                 160 





Cys Arg Ala Ser Gln Gly Ile Ser Ser Tyr Leu Ala Trp Tyr Gln Gln 

                165                 170                 175     





Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr Ala Ala Ser Thr Leu 

            180                 185                 190         





Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu 

        195                 200                 205             





Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr 

    210                 215                 220                 





Tyr Cys Gln Gln Leu Asn Ser Tyr Pro Pro Tyr Thr Phe Gly Gln Gly 

225                 230                 235                 240 





Thr Lys Leu Glu Ile Lys Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 

                245                 250                 255     





Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 

            260                 265                 270         





Gly Gly Gly Gly Ser Cys Ser Gly Gly Ser Gly Lys Asn Gly Ile Lys 

        275                 280                 285             





Val Asn Phe Lys Thr Arg His Asn Ile Glu Asp Gly Ser Val Gln Leu 

    290                 295                 300                 





Ala Asp His Tyr Gln Gln Asn Thr Pro Ile Gly Asp Gly Pro Val Leu 

305                 310                 315                 320 





Leu Pro Asp Asn His Tyr Leu Ser Thr Gln Ser Ala Leu Ser Lys Asp 

                325                 330                 335     





Pro Asn Glu Lys Arg Asp His Met Val Leu Leu Glu Phe Val Thr Ala 

            340                 345                 350         





Ala Gly Ile Thr His Gly Met Asp Glu Leu Tyr Asn 

        355                 360                 





<210>  6

<211>  483

<212>  PRT

<213>  Artificial Sequence



<220>

<223>  Split GFP 2 with COVID19 scfv



<400>  6



Met Gly Leu Asn Asp Ile Phe Glu Ala Gln Lys Ile Glu Trp His Glu 

1               5                   10                  15      





Gly Gly Gly Gly Ser Met Ala Ser Lys Gly Glu Glu Leu Phe Thr Gly 

            20                  25                  30          





Val Val Pro Ile Leu Val Glu Leu Asp Gly Asp Val Asn Gly His Lys 

        35                  40                  45              





Phe Ser Val Ser Gly Glu Gly Glu Gly Asp Ala Thr Tyr Gly Lys Leu 

    50                  55                  60                  





Thr Leu Lys Phe Ile Cys Thr Thr Gly Lys Leu Pro Val Pro Trp Pro 

65                  70                  75                  80  





Thr Leu Val Thr Thr Leu Cys Tyr Gly Val Gln Cys Phe Ser Arg Tyr 

                85                  90                  95      





Pro Asp His Met Lys Arg His Asp Phe Phe Lys Ser Ala Met Pro Glu 

            100                 105                 110         





Gly Tyr Val Gln Glu Arg Thr Ile Phe Phe Lys Asp Asp Gly Asn Tyr 

        115                 120                 125             





Lys Thr Arg Ala Glu Val Lys Phe Glu Gly Asp Thr Leu Val Asn Arg 

    130                 135                 140                 





Ile Glu Leu Lys Gly Ile Asp Phe Lys Glu Asp Gly Asn Ile Leu Gly 

145                 150                 155                 160 





His Lys Leu Glu Tyr Asn Tyr Asn His Asn Val Leu Ile Met Ala Asp 

                165                 170                 175     





Lys Gln Gly Gly Ser Gly Ser Gly Cys Ser Gly Gly Gly Gly Ser Gly 

            180                 185                 190         





Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 

        195                 200                 205             





Gly Gly Ser Gly Gly Gly Gly Ser Gly Met Gln Val Gln Leu Val Gln 

    210                 215                 220                 





Ser Gly Ala Glu Val Lys Lys Pro Gly Ala Ser Val Lys Val Ser Cys 

225                 230                 235                 240 





Lys Ala Ser Gly Tyr Thr Phe Thr Gly Tyr Tyr Met His Trp Val Arg 

                245                 250                 255     





Gln Ala Pro Gly Gln Gly Leu Glu Trp Met Gly Arg Ile Asn Pro Asn 

            260                 265                 270         





Ser Gly Gly Thr Asn Tyr Ala Gln Lys Phe Gln Gly Arg Val Thr Met 

        275                 280                 285             





Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr Met Glu Leu Ser Arg Leu 

    290                 295                 300                 





Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys Ala Arg Val Pro Tyr Cys 

305                 310                 315                 320 





Ser Ser Thr Ser Cys His Arg Asp Trp Tyr Phe Asp Leu Trp Gly Arg 

                325                 330                 335     





Gly Thr Leu Val Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly 

            340                 345                 350         





Gly Ser Gly Gly Gly Gly Ser Asp Ile Gln Met Thr Gln Ser Pro Leu 

        355                 360                 365             





Ser Leu Pro Val Thr Pro Gly Glu Pro Ala Ser Ile Ser Cys Arg Ser 

    370                 375                 380                 





Ser Gln Ser Leu Leu Asp Ser Asp Asp Gly Asn Thr Tyr Leu Asp Trp 

385                 390                 395                 400 





Tyr Leu Gln Lys Pro Gly Gln Ser Pro Gln Leu Leu Ile Tyr Thr Leu 

                405                 410                 415     





Ser Tyr Arg Ala Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser 

            420                 425                 430         





Gly Thr Asp Phe Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val 

        435                 440                 445             





Gly Val Tyr Tyr Cys Met Gln Arg Ile Glu Phe Pro Leu Thr Phe Gly 

    450                 455                 460                 





Gly Gly Thr Lys Val Glu Ile Lys Ser Gly Gly Gly Ser His His His 

465                 470                 475                 480 





His His His 

            





