                         SEQUENCE LISTING

<110>  New England Biolabs, Inc.
 
<120>  TARGET INITIATION AND AMPLIFICATION OF LONG DNA WITH NUCLEASE AND
        REPLISOME ENZYMES

<130>  NEB-439-PCT

<150>  63/208,534
<151>  2021-06-09

<150>  63208621
<151>  2021-06-09

<160>  2     

<170>  PatentIn version 3.5

<210>  1
<211>  564
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic construct

<400>  1

Asp Asn Ser His Asp Ser Asp Ser Val Phe Leu Tyr His Ile Pro Cys 
1               5                   10                  15      


Asp Asn Cys Gly Ser Ser Asp Gly Asn Ser Leu Phe Ser Asp Gly His 
            20                  25                  30          


Thr Phe Cys Tyr Val Cys Glu Lys Trp Thr Ala Gly Asn Glu Asp Thr 
        35                  40                  45              


Lys Glu Arg Ala Ser Lys Arg Lys Pro Ser Gly Gly Lys Pro Gly Thr 
    50                  55                  60                  


Tyr Asn Val Trp Asn Phe Gly Glu Ser Asn Gly Arg Tyr Ser Ala Leu 
65                  70                  75                  80  


Thr Ala Arg Gly Ile Ser Lys Glu Thr Cys Gln Lys Ala Gly Tyr Trp 
                85                  90                  95      


Ile Ala Lys Val Asp Gly Val Met Tyr Gln Val Ala Asp Tyr Arg Asp 
            100                 105                 110         


Gln Asn Gly Asn Ile Val Ser Gln Lys Val Arg Asp Lys Asp Lys Asn 
        115                 120                 125             


Phe Lys Thr Thr Gly Ser His Lys Ser Asp Ala Leu Phe Gly Lys His 
    130                 135                 140                 


Leu Trp Asn Gly Gly Lys Lys Ile Val Val Thr Glu Gly Glu Ile Asp 
145                 150                 155                 160 


Met Leu Thr Val Met Glu Leu Gln Asp Cys Lys Tyr Pro Val Val Ser 
                165                 170                 175     


Leu Gly His Gly Ala Ser Ala Ala Lys Lys Thr Cys Ala Ala Asn Tyr 
            180                 185                 190         


Glu Tyr Phe Asp Gln Phe Glu Gln Ile Ile Leu Met Phe Asp Met Asp 
        195                 200                 205             


Glu Ala Gly Arg Lys Ala Val Glu Glu Ala Ala Gln Val Leu Pro Ala 
    210                 215                 220                 


Gly Lys Val Arg Val Ala Val Leu Pro Cys Lys Asp Ala Asn Glu Cys 
225                 230                 235                 240 


His Leu Asn Gly His Asp Arg Glu Ile Met Glu Gln Val Trp Asn Ala 
                245                 250                 255     


Gly Pro Trp Ile Pro Asp Gly Val Val Ser Ala Leu Ser Leu Arg Glu 
            260                 265                 270         


Arg Ile Arg Glu His Leu Ser Ser Glu Glu Ser Val Gly Leu Leu Phe 
        275                 280                 285             


Ser Gly Cys Thr Gly Ile Asn Asp Lys Thr Leu Gly Ala Arg Gly Gly 
    290                 295                 300                 


Glu Val Ile Met Val Thr Ser Gly Ser Gly Met Gly Lys Ser Thr Phe 
305                 310                 315                 320 


Val Arg Gln Gln Ala Leu Gln Trp Gly Thr Ala Met Gly Lys Lys Val 
                325                 330                 335     


Gly Leu Ala Met Leu Glu Glu Ser Val Glu Glu Thr Ala Glu Asp Leu 
            340                 345                 350         


Ile Gly Leu His Asn Arg Val Arg Leu Arg Gln Ser Asp Ser Leu Lys 
        355                 360                 365             


Arg Glu Ile Ile Glu Asn Gly Lys Phe Asp Gln Trp Phe Asp Glu Leu 
    370                 375                 380                 


Phe Gly Asn Asp Thr Phe His Leu Tyr Asp Ser Phe Ala Glu Ala Glu 
385                 390                 395                 400 


Thr Asp Arg Leu Leu Ala Lys Leu Ala Tyr Met Arg Ser Gly Leu Gly 
                405                 410                 415     


Cys Asp Val Ile Ile Leu Asp His Ile Ser Ile Val Val Ser Ala Ser 
            420                 425                 430         


Gly Glu Ser Asp Glu Arg Lys Met Ile Asp Asn Leu Met Thr Lys Leu 
        435                 440                 445             


Lys Gly Phe Ala Lys Ser Thr Gly Val Val Leu Val Val Ile Cys His 
    450                 455                 460                 


Leu Lys Asn Pro Asp Lys Gly Lys Ala His Glu Glu Gly Arg Pro Val 
465                 470                 475                 480 


Ser Ile Thr Asp Leu Arg Gly Ser Gly Ala Leu Arg Gln Leu Ser Asp 
                485                 490                 495     


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


Val Leu Val Arg Ile Leu Lys Cys Arg Phe Thr Gly Asp Thr Gly Ile 
        515                 520                 525             


Ala Gly Tyr Met Glu Tyr Asn Lys Glu Thr Gly Trp Leu Glu Pro Ser 
    530                 535                 540                 


Ser Tyr Ser Gly Glu Glu Glu Ser His Ser Glu Ser Thr Asp Trp Ser 
545                 550                 555                 560 


Asn Asp Thr Asp 
                


<210>  2
<211>  502
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic construct

<400>  2

Thr Tyr Asn Val Trp Asn Phe Gly Glu Ser Asn Gly Arg Tyr Ser Ala 
1               5                   10                  15      


Leu Thr Ala Arg Gly Ile Ser Lys Glu Thr Cys Gln Lys Ala Gly Tyr 
            20                  25                  30          


Trp Ile Ala Lys Val Asp Gly Val Met Tyr Gln Val Ala Asp Tyr Arg 
        35                  40                  45              


Asp Gln Asn Gly Asn Ile Val Ser Gln Lys Val Arg Asp Lys Asp Lys 
    50                  55                  60                  


Asn Phe Lys Thr Thr Gly Ser His Lys Ser Asp Ala Leu Phe Gly Lys 
65                  70                  75                  80  


His Leu Trp Asn Gly Gly Lys Lys Ile Val Val Thr Glu Gly Glu Ile 
                85                  90                  95      


Asp Met Leu Thr Val Met Glu Leu Gln Asp Cys Lys Tyr Pro Val Val 
            100                 105                 110         


Ser Leu Gly His Gly Ala Ser Ala Ala Lys Lys Thr Cys Ala Ala Asn 
        115                 120                 125             


Tyr Glu Tyr Phe Asp Gln Phe Glu Gln Ile Ile Leu Met Phe Asp Met 
    130                 135                 140                 


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


Ala Gly Lys Val Arg Val Ala Val Leu Pro Cys Lys Asp Ala Asn Glu 
                165                 170                 175     


Cys His Leu Asn Gly His Asp Arg Glu Ile Met Glu Gln Val Trp Asn 
            180                 185                 190         


Ala Gly Pro Trp Ile Pro Asp Gly Val Val Ser Ala Leu Ser Leu Arg 
        195                 200                 205             


Glu Arg Ile Arg Glu His Leu Ser Ser Glu Glu Ser Val Gly Leu Leu 
    210                 215                 220                 


Phe Ser Gly Cys Thr Gly Ile Asn Asp Lys Thr Leu Gly Ala Arg Gly 
225                 230                 235                 240 


Gly Glu Val Ile Met Val Thr Ser Gly Ser Gly Met Gly Lys Ser Thr 
                245                 250                 255     


Phe Val Arg Gln Gln Ala Leu Gln Trp Gly Thr Ala Met Gly Lys Lys 
            260                 265                 270         


Val Gly Leu Ala Met Leu Glu Glu Ser Val Glu Glu Thr Ala Glu Asp 
        275                 280                 285             


Leu Ile Gly Leu His Asn Arg Val Arg Leu Arg Gln Ser Asp Ser Leu 
    290                 295                 300                 


Lys Arg Glu Ile Ile Glu Asn Gly Lys Phe Asp Gln Trp Phe Asp Glu 
305                 310                 315                 320 


Leu Phe Gly Asn Asp Thr Phe His Leu Tyr Asp Ser Phe Ala Glu Ala 
                325                 330                 335     


Glu Thr Asp Arg Leu Leu Ala Lys Leu Ala Tyr Met Arg Ser Gly Leu 
            340                 345                 350         


Gly Cys Asp Val Ile Ile Leu Asp His Ile Ser Ile Val Val Ser Ala 
        355                 360                 365             


Ser Gly Glu Ser Asp Glu Arg Lys Met Ile Asp Asn Leu Met Thr Lys 
    370                 375                 380                 


Leu Lys Gly Phe Ala Lys Ser Thr Gly Val Val Leu Val Val Ile Cys 
385                 390                 395                 400 


His Leu Lys Asn Pro Asp Lys Gly Lys Ala His Glu Glu Gly Arg Pro 
                405                 410                 415     


Val Ser Ile Thr Asp Leu Arg Gly Ser Gly Ala Leu Arg Gln Leu Ser 
            420                 425                 430         


Asp Thr Ile Ile Ala Leu Glu Arg Asn Gln Gln Gly Asp Met Pro Asn 
        435                 440                 445             


Leu Val Leu Val Arg Ile Leu Lys Cys Arg Phe Thr Gly Asp Thr Gly 
    450                 455                 460                 


Ile Ala Gly Tyr Met Glu Tyr Asn Lys Glu Thr Gly Trp Leu Glu Pro 
465                 470                 475                 480 


Ser Ser Tyr Ser Gly Glu Glu Glu Ser His Ser Glu Ser Thr Asp Trp 
                485                 490                 495     


Ser Asn Asp Thr Asp Phe 
            500         


