                         SEQUENCE LISTING

<110>  The General Hospital Corporation
 
<120>  Methods and Compositions for Enzymatic Polymerization of N3' -> 
       P5' Phosphoramidate DNA

<130>  29539-0479WO1

<150>  US 62/939,724
<151>  2019-11-25

<160>  7     

<170>  PatentIn version 3.5

<210>  1
<211>  876
<212>  PRT
<213>  Bacillus stearothermophilus

<400>  1

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


Ala Phe Phe Ala Leu Pro Leu Leu His Asn Asp Lys Gly Ile His Thr 
            20                  25                  30          


Asn Ala Val Tyr Gly Phe Thr Met Met Leu Asn Lys Ile Leu Ala Glu 
        35                  40                  45              


Glu Gln Pro Thr His Ile Leu Val Ala Phe Asp Ala Gly Lys Thr Thr 
    50                  55                  60                  


Phe Arg His Glu Thr Phe Gln Asp Tyr Lys Gly Gly Arg Gln Gln Thr 
65                  70                  75                  80  


Pro Pro Glu Leu Ser Glu Gln Phe Pro Leu Leu Arg Glu Leu Leu Lys 
                85                  90                  95      


Ala Tyr Arg Ile Pro Ala Tyr Glu Leu Asp His Tyr Glu Ala Asp Asp 
            100                 105                 110         


Ile Ile Gly Thr Met Ala Ala Arg Ala Glu Arg Glu Gly Phe Ala Val 
        115                 120                 125             


Lys Val Ile Ser Gly Asp Arg Asp Leu Thr Gln Leu Ala Ser Pro Gln 
    130                 135                 140                 


Val Thr Val Glu Ile Thr Lys Lys Gly Ile Thr Asp Ile Glu Ser Tyr 
145                 150                 155                 160 


Thr Pro Glu Thr Val Val Glu Lys Tyr Gly Leu Thr Pro Glu Gln Ile 
                165                 170                 175     


Val Asp Leu Lys Gly Leu Met Gly Asp Lys Ser Asp Asn Ile Pro Gly 
            180                 185                 190         


Val Pro Gly Ile Gly Glu Lys Thr Ala Val Lys Leu Leu Lys Gln Phe 
        195                 200                 205             


Gly Thr Val Glu Asn Val Leu Ala Ser Ile Asp Glu Ile Lys Gly Glu 
    210                 215                 220                 


Lys Leu Lys Glu Asn Leu Arg Gln Tyr Arg Asp Leu Ala Leu Leu Ser 
225                 230                 235                 240 


Lys Gln Leu Ala Ala Ile Cys Arg Asp Ala Pro Val Glu Leu Thr Leu 
                245                 250                 255     


Asp Asp Ile Val Tyr Lys Gly Glu Asp Arg Glu Lys Val Val Ala Leu 
            260                 265                 270         


Phe Gln Glu Leu Gly Phe Gln Ser Phe Leu Asp Lys Met Ala Val Gln 
        275                 280                 285             


Thr Asp Glu Gly Glu Lys Pro Leu Ala Gly Met Asp Phe Ala Ile Ala 
    290                 295                 300                 


Asp Ser Val Thr Asp Glu Met Leu Ala Asp Lys Ala Ala Leu Val Val 
305                 310                 315                 320 


Glu Val Val Gly Asp Asn Tyr His His Ala Pro Ile Val Gly Ile Ala 
                325                 330                 335     


Leu Ala Asn Glu Arg Gly Arg Phe Phe Leu Arg Pro Glu Thr Ala Leu 
            340                 345                 350         


Ala Asp Pro Lys Phe Leu Ala Trp Leu Gly Asp Glu Thr Lys Lys Lys 
        355                 360                 365             


Thr Met Phe Asp Ser Lys Arg Ala Ala Val Ala Leu Lys Trp Lys Gly 
    370                 375                 380                 


Ile Glu Leu Arg Gly Val Val Phe Asp Leu Leu Leu Ala Ala Tyr Leu 
385                 390                 395                 400 


Leu Asp Pro Ala Gln Ala Ala Gly Asp Val Ala Ala Val Ala Lys Met 
                405                 410                 415     


His Gln Tyr Glu Ala Val Arg Ser Asp Glu Ala Val Tyr Gly Lys Gly 
            420                 425                 430         


Ala Lys Arg Thr Val Pro Asp Glu Pro Thr Leu Ala Glu His Leu Val 
        435                 440                 445             


Arg Lys Ala Ala Ala Ile Trp Ala Leu Glu Glu Pro Leu Met Asp Glu 
    450                 455                 460                 


Leu Arg Arg Asn Glu Gln Asp Arg Leu Leu Thr Glu Leu Glu Gln Pro 
465                 470                 475                 480 


Leu Ala Gly Ile Leu Ala Asn Met Glu Phe Thr Gly Val Lys Val Asp 
                485                 490                 495     


Thr Lys Arg Leu Glu Gln Met Gly Ala Glu Leu Thr Glu Gln Leu Gln 
            500                 505                 510         


Ala Val Glu Arg Arg Ile Tyr Glu Leu Ala Gly Gln Glu Phe Asn Ile 
        515                 520                 525             


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


Pro Val Leu Lys Lys Thr Lys Thr Gly Tyr Ser Thr Ser Ala Asp Val 
545                 550                 555                 560 


Leu Glu Lys Leu Ala Pro His His Glu Ile Val Glu His Ile Leu His 
                565                 570                 575     


Tyr Arg Gln Leu Gly Lys Leu Gln Ser Thr Tyr Ile Glu Gly Leu Leu 
            580                 585                 590         


Lys Val Val His Pro Val Thr Gly Lys Val His Thr Met Phe Asn Gln 
        595                 600                 605             


Ala Leu Thr Gln Thr Gly Arg Leu Ser Ser Val Glu Pro Asn Leu Gln 
    610                 615                 620                 


Asn Ile Pro Ile Arg Leu Glu Glu Gly Arg Lys Ile Arg Gln Ala Phe 
625                 630                 635                 640 


Val Pro Ser Glu Pro Asp Trp Leu Ile Phe Ala Ala Asp Tyr Ser Gln 
                645                 650                 655     


Ile Glu Leu Arg Val Leu Ala His Ile Ala Glu Asp Asp Asn Leu Ile 
            660                 665                 670         


Glu Ala Phe Arg Arg Gly Leu Asp Ile His Thr Lys Thr Ala Met Asp 
        675                 680                 685             


Ile Phe His Val Ser Glu Glu Asp Val Thr Ala Asn Met Arg Arg Gln 
    690                 695                 700                 


Ala Lys Ala Val Asn Phe Gly Ile Val Tyr Gly Ile Ser Asp Tyr Gly 
705                 710                 715                 720 


Leu Ala Gln Asn Leu Asn Ile Thr Arg Lys Glu Ala Ala Glu Phe Ile 
                725                 730                 735     


Glu Arg Tyr Phe Ala Ser Phe Pro Gly Val Lys Gln Tyr Met Asp Asn 
            740                 745                 750         


Ile Val Gln Glu Ala Lys Gln Lys Gly Tyr Val Thr Thr Leu Leu His 
        755                 760                 765             


Arg Arg Arg Tyr Leu Pro Asp Ile Thr Ser Arg Asn Phe Asn Val Arg 
    770                 775                 780                 


Ser Phe Ala Glu Arg Thr Ala Met Asn Thr Pro Ile Gln Gly Ser Ala 
785                 790                 795                 800 


Ala Asp Ile Ile Lys Lys Ala Met Ile Asp Leu Ser Val Arg Leu Arg 
                805                 810                 815     


Glu Glu Arg Leu Gln Ala Arg Leu Leu Leu Gln Val His Asp Glu Leu 
            820                 825                 830         


Ile Leu Glu Ala Pro Lys Glu Glu Ile Glu Arg Leu Cys Arg Leu Val 
        835                 840                 845             


Pro Glu Val Met Glu Gln Ala Val Thr Leu Arg Val Pro Leu Lys Val 
    850                 855                 860                 


Asp Tyr His Tyr Gly Pro Thr Trp Tyr Asp Ala Lys 
865                 870                 875     


<210>  2
<211>  9
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic primer


<220>
<221>  modified_base
<222>  (9)..(9)
<223>  3'-amino-2',3'-dideoxycytidine (nC) or 2',3'-Dideoxycytidine 
       (ddC)

<400>  2
gcgatcagc                                                                9


<210>  3
<211>  14
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic primer

<400>  3
acacgctgat cgca                                                         14


<210>  4
<211>  18
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  3'-amino terminal DNA primer

<400>  4
gcgtccattc gttacctg                                                     18


<210>  5
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic template

<400>  5
cgcaggtaag caatggacgg act                                               23


<210>  6
<211>  46
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synethetic template

<400>  6
cgcaggtaag caatggacgg actacgcact cataagttgt aaaggc                      46


<210>  7
<211>  24
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  synthetic template

<400>  7
cgcgaattcg cgcgcgaatt cgcg                                              24


