                         SEQUENCE LISTING

<110>  New England Biolabs, Inc.
 
<120>  AN ISOTHERMAL DIAGNOSTIC TEST THAT UTILIZES A CAS PROTEIN AND A 
       POLYMERASE

<130>  NEB-447-PCT

<150>  63/209,008
<151>  2021-06-09

<160>  10    

<170>  PatentIn version 3.5

<210>  1
<211>  584
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic construct

<400>  1

Asp Gln Ser Leu Glu Asp Ile Asn Val Lys Thr Val Thr Asp Val Thr 
1               5                   10                  15      


Ser Asp Ile Leu Val Ser Pro Ser Ala Phe Val Val Glu Gln Ile Gly 
            20                  25                  30          


Asp Asn Tyr His Glu Glu Pro Ile Leu Gly Phe Ser Ile Val Asn Glu 
        35                  40                  45              


Thr Gly Ala Tyr Phe Ile Pro Lys Asp Ile Ala Val Glu Ser Glu Val 
    50                  55                  60                  


Phe Lys Glu Trp Val Glu Asn Asp Glu Gln Lys Lys Trp Val Phe Asp 
65                  70                  75                  80  


Ser Lys Arg Ala Val Val Ala Leu Arg Trp Gln Gly Ile Glu Leu Lys 
                85                  90                  95      


Gly Ala Glu Phe Asp Thr Leu Leu Ala Ala Tyr Ile Ile Asn Pro Gly 
            100                 105                 110         


Asn Ser Tyr Asp Asp Val Ala Ser Val Ala Lys Asp Tyr Gly Leu His 
        115                 120                 125             


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


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


Ala Ile Gln Ser Leu Arg Glu Lys Leu Val Gln Glu Leu Glu Asn Asn 
                165                 170                 175     


Asp Gln Leu Glu Leu Phe Glu Glu Leu Glu Met Pro Leu Ala Leu Ile 
            180                 185                 190         


Leu Gly Glu Met Glu Ser Thr Gly Val Lys Val Asp Val Asp Arg Leu 
        195                 200                 205             


Lys Arg Met Gly Glu Glu Leu Gly Ala Lys Leu Lys Glu Tyr Glu Glu 
    210                 215                 220                 


Lys Ile His Glu Ile Ala Gly Glu Pro Phe Asn Ile Asn Ser Pro Lys 
225                 230                 235                 240 


Gln Leu Gly Val Ile Leu Phe Glu Lys Ile Gly Leu Pro Val Val Lys 
                245                 250                 255     


Lys Thr Lys Thr Gly Tyr Ser Thr Ser Ala Asp Val Leu Glu Lys Leu 
            260                 265                 270         


Ala Asp Lys His Asp Ile Val Asp Tyr Ile Leu Gln Tyr Arg Gln Ile 
        275                 280                 285             


Gly Lys Leu Gln Ser Thr Tyr Ile Glu Gly Leu Leu Lys Val Thr Arg 
    290                 295                 300                 


Pro Asp Ser His Lys Val His Thr Arg Phe Asn Gln Ala Leu Thr Gln 
305                 310                 315                 320 


Thr Gly Arg Leu Ser Ser Thr Asp Pro Asn Leu Gln Asn Ile Pro Ile 
                325                 330                 335     


Arg Leu Glu Glu Gly Arg Lys Ile Arg Gln Ala Phe Val Pro Ser Glu 
            340                 345                 350         


Lys Asp Trp Leu Ile Phe Ala Ala Asp Tyr Ser Gln Ile Glu Leu Arg 
        355                 360                 365             


Val Leu Ala His Ile Ser Lys Asp Glu Asn Leu Ile Glu Ala Phe Thr 
    370                 375                 380                 


Asn Asp Met Asp Ile His Thr Lys Thr Ala Met Asp Val Phe His Val 
385                 390                 395                 400 


Ala Lys Asp Glu Val Thr Ser Ala Met Arg Arg Gln Ala Lys Ala Val 
                405                 410                 415     


Asn Phe Gly Ile Val Tyr Gly Ile Ser Asp Tyr Gly Leu Ser Gln Asn 
            420                 425                 430         


Leu Gly Ile Thr Arg Lys Glu Ala Gly Ala Phe Ile Asp Arg Tyr Leu 
        435                 440                 445             


Glu Ser Phe Gln Gly Val Lys Ala Tyr Met Glu Asp Ser Val Gln Glu 
    450                 455                 460                 


Ala Lys Gln Lys Gly Tyr Val Thr Thr Leu Met His Arg Arg Arg Tyr 
465                 470                 475                 480 


Ile Pro Glu Leu Thr Ser Arg Asn Phe Asn Ile Arg Ser Phe Ala Glu 
                485                 490                 495     


Arg Thr Ala Met Asn Thr Pro Ile Gln Gly Ser Ala Ala Asp Ile Ile 
            500                 505                 510         


Lys Lys Ala Met Ile Asp Met Ala Ala Lys Leu Lys Glu Lys Gln Leu 
        515                 520                 525             


Lys Ala Arg Leu Leu Leu Gln Val His Asp Glu Leu Ile Phe Glu Ala 
    530                 535                 540                 


Pro Lys Glu Glu Ile Glu Ile Leu Glu Lys Leu Val Pro Glu Val Met 
545                 550                 555                 560 


Glu His Ala Leu Ala Leu Asp Val Pro Leu Lys Val Asp Phe Ala Ser 
                565                 570                 575     


Gly Pro Ser Trp Tyr Asp Ala Lys 
            580                 


<210>  2
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic construct

<400>  2
tgagtacgaa cttatgtact cat                                               23


<210>  3
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic construct

<400>  3
ttcagatttt taacacgaga gt                                                22


<210>  4
<211>  42
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic construct

<400>  4
accacgaaag caagaaaaag aagttcgttt cggaagagac ag                          42


<210>  5
<211>  44
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic construct

<400>  5
ttgctagtta cactagccat ccttaggttt tacaagactc acgt                        44


<210>  6
<211>  19
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic construct

<400>  6
gcgcttcgat tgtgtgcgt                                                    19


<210>  7
<211>  18
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic construct

<400>  7
cgctattaac tattaacg                                                     18


<210>  8
<211>  41
<212>  RNA
<213>  Artificial Sequence

<220>
<223>  Synthetic construct

<400>  8
uaauuucuac uaaguguaga ugugguauuc uugcuaguua c                           41


<210>  9
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic construct

<400>  9
cagtcagtca gtcagtcagt                                                   20


<210>  10
<211>  1246
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic construct

<400>  10

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


Lys Asn Met Ser Lys Leu Glu Lys Phe Thr Asn Cys Tyr Ser Leu Ser 
            20                  25                  30          


Lys Thr Leu Arg Phe Lys Ala Ile Pro Val Gly Lys Thr Gln Glu Asn 
        35                  40                  45              


Ile Asp Asn Lys Arg Leu Leu Val Glu Asp Glu Lys Arg Ala Glu Asp 
    50                  55                  60                  


Tyr Lys Gly Val Lys Lys Leu Leu Asp Arg Tyr Tyr Leu Ser Phe Ile 
65                  70                  75                  80  


Asn Asp Val Leu His Ser Ile Lys Leu Lys Asn Leu Asn Asn Tyr Ile 
                85                  90                  95      


Ser Leu Phe Arg Lys Lys Thr Arg Thr Glu Lys Glu Asn Lys Glu Leu 
            100                 105                 110         


Glu Asn Leu Glu Ile Asn Leu Arg Lys Glu Ile Ala Lys Ala Phe Lys 
        115                 120                 125             


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


Ile Leu Pro Glu Phe Leu Asp Asp Lys Asp Glu Ile Ala Leu Val Asn 
145                 150                 155                 160 


Ser Phe Asn Gly Phe Thr Thr Ala Phe Thr Gly Phe Phe Asp Asn Arg 
                165                 170                 175     


Glu Asn Met Phe Ser Glu Glu Ala Lys Ser Thr Ser Ile Ala Phe Arg 
            180                 185                 190         


Cys Ile Asn Glu Asn Leu Thr Arg Tyr Ile Ser Asn Met Asp Ile Phe 
        195                 200                 205             


Glu Lys Val Asp Ala Ile Phe Asp Lys His Glu Val Gln Glu Ile Lys 
    210                 215                 220                 


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


Glu Phe Phe Asn Phe Val Leu Thr Gln Glu Gly Ile Asp Val Tyr Asn 
                245                 250                 255     


Ala Ile Ile Gly Gly Phe Val Thr Glu Ser Gly Glu Lys Ile Lys Gly 
            260                 265                 270         


Leu Asn Glu Tyr Ile Asn Leu Tyr Asn Gln Lys Thr Lys Gln Lys Leu 
        275                 280                 285             


Pro Lys Phe Lys Pro Leu Tyr Lys Gln Val Leu Ser Asp Arg Glu Ser 
    290                 295                 300                 


Leu Ser Phe Tyr Gly Glu Gly Tyr Thr Ser Asp Glu Glu Val Leu Glu 
305                 310                 315                 320 


Val Phe Arg Asn Thr Leu Asn Lys Asn Ser Glu Ile Phe Ser Ser Ile 
                325                 330                 335     


Lys Lys Leu Glu Lys Leu Phe Lys Asn Phe Asp Glu Tyr Ser Ser Ala 
            340                 345                 350         


Gly Ile Phe Val Lys Asn Gly Pro Ala Ile Ser Thr Ile Ser Lys Asp 
        355                 360                 365             


Ile Phe Gly Glu Trp Asn Val Ile Arg Asp Lys Trp Asn Ala Glu Tyr 
    370                 375                 380                 


Asp Asp Ile His Leu Lys Lys Lys Ala Val Val Thr Glu Lys Tyr Glu 
385                 390                 395                 400 


Asp Asp Arg Arg Lys Ser Phe Lys Lys Ile Gly Ser Phe Ser Leu Glu 
                405                 410                 415     


Gln Leu Gln Glu Tyr Ala Asp Ala Asp Leu Ser Val Val Glu Lys Leu 
            420                 425                 430         


Lys Glu Ile Ile Ile Gln Lys Val Asp Glu Ile Tyr Lys Val Tyr Gly 
        435                 440                 445             


Ser Ser Glu Lys Leu Phe Asp Ala Asp Phe Val Leu Glu Lys Ser Leu 
    450                 455                 460                 


Lys Lys Asn Asp Ala Val Val Ala Ile Met Lys Asp Leu Leu Asp Ser 
465                 470                 475                 480 


Val Lys Ser Phe Glu Asn Tyr Ile Lys Ala Phe Phe Gly Glu Gly Lys 
                485                 490                 495     


Glu Thr Asn Arg Asp Glu Ser Phe Tyr Gly Asp Phe Val Leu Ala Tyr 
            500                 505                 510         


Asp Ile Leu Leu Lys Val Asp His Ile Tyr Asp Ala Ile Arg Asn Tyr 
        515                 520                 525             


Val Thr Gln Lys Pro Tyr Ser Lys Asp Lys Phe Lys Leu Tyr Phe Gln 
    530                 535                 540                 


Asn Pro Gln Phe Met Gly Gly Trp Asp Lys Asp Lys Glu Thr Asp Tyr 
545                 550                 555                 560 


Arg Ala Thr Ile Leu Arg Tyr Gly Ser Lys Tyr Tyr Leu Ala Ile Met 
                565                 570                 575     


Asp Lys Lys Tyr Ala Lys Cys Leu Gln Lys Ile Asp Lys Asp Asp Val 
            580                 585                 590         


Asn Gly Asn Tyr Glu Lys Ile Asn Tyr Lys Leu Leu Pro Gly Pro Asn 
        595                 600                 605             


Lys Met Leu Pro Lys Val Phe Phe Ser Lys Lys Trp Met Ala Tyr Tyr 
    610                 615                 620                 


Asn Pro Ser Glu Asp Ile Gln Lys Ile Tyr Lys Asn Gly Thr Phe Lys 
625                 630                 635                 640 


Lys Gly Asp Met Phe Asn Leu Asn Asp Cys His Lys Leu Ile Asp Phe 
                645                 650                 655     


Phe Lys Asp Ser Ile Ser Arg Tyr Pro Lys Trp Ser Asn Ala Tyr Asp 
            660                 665                 670         


Phe Asn Phe Ser Glu Thr Glu Lys Tyr Lys Asp Ile Ala Gly Phe Tyr 
        675                 680                 685             


Arg Glu Val Glu Glu Gln Gly Tyr Lys Val Ser Phe Glu Ser Ala Ser 
    690                 695                 700                 


Lys Lys Glu Val Asp Lys Leu Val Glu Glu Gly Lys Leu Tyr Met Phe 
705                 710                 715                 720 


Gln Ile Tyr Asn Lys Asp Phe Ser Asp Lys Ser His Gly Thr Pro Asn 
                725                 730                 735     


Leu His Thr Met Tyr Phe Lys Leu Leu Phe Asp Glu Asn Asn His Gly 
            740                 745                 750         


Gln Ile Arg Leu Ser Gly Gly Ala Glu Leu Phe Met Arg Arg Ala Ser 
        755                 760                 765             


Leu Lys Lys Glu Glu Leu Val Val His Pro Ala Asn Ser Pro Ile Ala 
    770                 775                 780                 


Asn Lys Asn Pro Asp Asn Pro Lys Lys Thr Thr Thr Leu Ser Tyr Asp 
785                 790                 795                 800 


Val Tyr Lys Asp Lys Arg Phe Ser Glu Asp Gln Tyr Glu Leu His Ile 
                805                 810                 815     


Pro Ile Ala Ile Asn Lys Cys Pro Lys Asn Ile Phe Lys Ile Asn Thr 
            820                 825                 830         


Glu Val Arg Val Leu Leu Lys His Asp Asp Asn Pro Tyr Val Ile Gly 
        835                 840                 845             


Ile Asp Arg Gly Glu Arg Asn Leu Leu Tyr Ile Val Val Val Asp Gly 
    850                 855                 860                 


Lys Gly Asn Ile Val Glu Gln Tyr Ser Leu Asn Glu Ile Ile Asn Asn 
865                 870                 875                 880 


Phe Asn Gly Ile Arg Ile Lys Thr Asp Tyr His Ser Leu Leu Asp Lys 
                885                 890                 895     


Lys Glu Lys Glu Arg Phe Glu Ala Arg Gln Asn Trp Thr Ser Ile Glu 
            900                 905                 910         


Asn Ile Lys Glu Leu Lys Ala Gly Tyr Ile Ser Gln Val Val His Lys 
        915                 920                 925             


Ile Cys Glu Leu Val Glu Lys Tyr Asp Ala Val Ile Ala Leu Glu Asp 
    930                 935                 940                 


Leu Asn Ser Gly Phe Lys Asn Ser Arg Val Lys Val Glu Lys Gln Val 
945                 950                 955                 960 


Tyr Gln Lys Phe Glu Lys Met Leu Ile Asp Lys Leu Asn Tyr Met Val 
                965                 970                 975     


Asp Lys Lys Ser Asn Pro Cys Ala Thr Gly Gly Ala Leu Lys Gly Tyr 
            980                 985                 990         


Gln Ile Thr Asn Lys Phe Glu Ser  Phe Lys Ser Met Ser  Thr Gln Asn 
        995                 1000                 1005             


Gly Phe  Ile Phe Tyr Ile Pro  Ala Trp Leu Thr Ser  Lys Ile Asp 
    1010                 1015                 1020             


Pro Ser  Thr Gly Phe Val Asn  Leu Leu Lys Thr Lys  Tyr Thr Ser 
    1025                 1030                 1035             


Ile Ala  Asp Ser Lys Lys Phe  Ile Ser Ser Phe Asp  Arg Ile Met 
    1040                 1045                 1050             


Tyr Val  Pro Glu Glu Asp Leu  Phe Glu Phe Ala Leu  Asp Tyr Lys 
    1055                 1060                 1065             


Asn Phe  Ser Arg Thr Asp Ala  Asp Tyr Ile Lys Lys  Trp Lys Leu 
    1070                 1075                 1080             


Tyr Ser  Tyr Gly Asn Arg Ile  Arg Ile Phe Arg Asn  Pro Lys Lys 
    1085                 1090                 1095             


Asn Asn  Val Phe Asp Trp Glu  Glu Val Cys Leu Thr  Ser Ala Tyr 
    1100                 1105                 1110             


Lys Glu  Leu Phe Asn Lys Tyr  Gly Ile Asn Tyr Gln  Gln Gly Asp 
    1115                 1120                 1125             


Ile Arg  Ala Leu Leu Cys Glu  Gln Ser Asp Lys Ala  Phe Tyr Ser 
    1130                 1135                 1140             


Ser Phe  Met Ala Leu Met Ser  Leu Met Leu Gln Met  Arg Asn Ser 
    1145                 1150                 1155             


Ile Thr  Gly Arg Thr Asp Val  Asp Phe Leu Ile Ser  Pro Val Lys 
    1160                 1165                 1170             


Asn Ser  Asp Gly Ile Phe Tyr  Asp Ser Arg Asn Tyr  Glu Ala Gln 
    1175                 1180                 1185             


Glu Asn  Ala Ile Leu Pro Lys  Asn Ala Asp Ala Asn  Gly Ala Tyr 
    1190                 1195                 1200             


Asn Ile  Ala Arg Lys Val Leu  Trp Ala Ile Gly Gln  Phe Lys Lys 
    1205                 1210                 1215             


Ala Glu  Asp Glu Lys Leu Asp  Lys Val Lys Ile Ala  Ile Ser Asn 
    1220                 1225                 1230             


Lys Glu  Trp Leu Glu Tyr Ala  Gln Thr Ser Val Lys  His 
    1235                 1240                 1245     


