                         LISTADO DE SECUENCIAS

<110>  UNIVERSIDAD DE CANTABRIA
       INSTITUT PASTEUR
       CONSEJO SUPERIOR DE INVESTIGACIONES CIENTFICAS
 
<120>  PROTENA QUIMRICA RELAXASACas

<130>  PCT3120.10

<150> ES P202030890
<151> 2020-08-28

<160>  26    

<170>  PatentIn version 3.5

<210>  1
<211>  2318
<212>  PRT
<213>  Secuencia Artificial

<220>
<223>  TrwC:Cas12a protena quimrica

<400>  1

Met Leu Ser His Met Val Leu Thr Arg Gln Asp Ile Gly Arg Ala Ala 
1               5                   10                  15      


Ser Tyr Tyr Glu Asp Gly Ala Asp Asp Tyr Tyr Ala Lys Asp Gly Asp 
            20                  25                  30          


Ala Ser Glu Trp Gln Gly Lys Gly Ala Glu Glu Leu Gly Leu Ser Gly 
        35                  40                  45              


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


Glu Gly His Arg Ile Met Arg Ser Ala Thr Arg Gln Asp Ser Lys Glu 
65                  70                  75                  80  


Arg Ile Gly Leu Asp Leu Thr Phe Ser Ala Pro Lys Ser Val Ser Leu 
                85                  90                  95      


Gln Ala Leu Val Ala Gly Asp Ala Glu Ile Ile Lys Ala His Asp Arg 
            100                 105                 110         


Ala Val Ala Arg Thr Leu Glu Gln Ala Glu Ala Arg Ala Gln Ala Arg 
        115                 120                 125             


Gln Lys Ile Gln Gly Lys Thr Arg Ile Glu Thr Thr Gly Asn Leu Val 
    130                 135                 140                 


Ile Gly Lys Phe Arg His Glu Thr Ser Arg Glu Arg Asp Pro Gln Leu 
145                 150                 155                 160 


His Thr His Ala Val Ile Leu Asn Met Thr Lys Arg Ser Asp Gly Gln 
                165                 170                 175     


Trp Arg Ala Leu Lys Asn Asp Glu Ile Val Lys Ala Thr Arg Tyr Leu 
            180                 185                 190         


Gly Ala Val Tyr Asn Ala Glu Leu Ala His Glu Leu Gln Lys Leu Gly 
        195                 200                 205             


Tyr Gln Leu Arg Tyr Gly Lys Asp Gly Asn Phe Asp Leu Ala His Ile 
    210                 215                 220                 


Asp Arg Gln Gln Ile Glu Gly Phe Ser Lys Arg Thr Glu Gln Ile Ala 
225                 230                 235                 240 


Glu Trp Tyr Ala Ala Arg Gly Leu Asp Pro Asn Ser Val Ser Leu Glu 
                245                 250                 255     


Gln Lys Gln Ala Ala Lys Val Leu Ser Arg Ala Lys Lys Thr Ser Val 
            260                 265                 270         


Asp Arg Glu Ala Leu Arg Ala Glu Trp Gln Ala Thr Ala Lys Glu Leu 
        275                 280                 285             


Gly Ile Asp Phe Ser Arg Arg Glu Trp Ser Gly Arg Glu Lys Gly Gly 
    290                 295                 300                 


Ser Glu Lys Gln Ala His Ser Phe Met Pro Ser Asp Glu Ala Ala Lys 
305                 310                 315                 320 


Arg Ala Val Arg Tyr Ala Ile Asn His Leu Thr Glu Arg Gln Ser Val 
                325                 330                 335     


Met Asp Glu Arg Glu Leu Val Asp Thr Ala Met Lys His Ala Val Gly 
            340                 345                 350         


Ala Ala Arg Leu Glu Asp Ile Gln Lys Glu Leu Leu Arg Gln Thr Glu 
        355                 360                 365             


Thr Gly Tyr Leu Ile Arg Glu Ala Pro Arg Tyr Arg Pro Gly Gly Gln 
    370                 375                 380                 


Thr Gly Pro Thr Asp Glu Pro Gly Lys Thr Arg Ala Glu Trp Val Ala 
385                 390                 395                 400 


Glu Leu Ala Ala Lys Gly Met Lys Gln Gly Ala Ala Arg Glu Arg Val 
                405                 410                 415     


Asp Asn Ala Ile Lys Thr Gly Gly Leu Val Pro Ile Glu Pro Arg Tyr 
            420                 425                 430         


Thr Thr Gln Thr Ala Leu Glu Arg Glu Lys Arg Ile Leu Gln Ile Glu 
        435                 440                 445             


Arg Asp Gly Arg Gly Ala Val Ala Pro Val Ile Ala Ala Glu Ala Ala 
    450                 455                 460                 


Arg Glu Arg Leu Ala Ser Thr Asn Leu Asn Gln Gly Gln Arg Glu Ala 
465                 470                 475                 480 


Ala Glu Leu Ile Val Ser Ala Ala Asn Arg Val Val Gly Val Gln Gly 
                485                 490                 495     


Phe Ala Gly Thr Gly Lys Ser His Met Leu Asp Thr Ala Lys Gln Met 
            500                 505                 510         


Ile Glu Gly Glu Gly Tyr His Val Arg Ala Leu Ala Ala Tyr Gly Ser 
        515                 520                 525             


Gln Val Lys Ala Leu Arg Glu Leu Asn Val Glu Ala Asn Thr Leu Ala 
    530                 535                 540                 


Ser Phe Leu Arg Ala Lys Asp Lys Asn Ile Asp Ser Arg Thr Val Leu 
545                 550                 555                 560 


Val Ile Asp Glu Ala Gly Val Val Pro Thr Arg Leu Met Glu Gln Thr 
                565                 570                 575     


Leu Lys Leu Ala Glu Lys Ala Gly Ala Arg Val Val Leu Met Gly Asp 
            580                 585                 590         


Thr Ala Gln Thr Lys Ala Ile Glu Ala Gly Arg Pro Phe Asp Gln Leu 
        595                 600                 605             


Gln Ala Ala Gly Met Gln Thr Ala His Met Arg Glu Ile Gln Arg Gln 
    610                 615                 620                 


Lys Asn Pro Glu Leu Lys Ile Ala Val Glu Leu Ala Ala Ala Gly Lys 
625                 630                 635                 640 


Ala Ser Ser Ser Leu Glu Arg Ile Lys Asp Val Thr Glu Ile Lys Asn 
                645                 650                 655     


His His Glu Arg Arg Ala Ala Val Ala Glu Ala Tyr Ile Ala Leu Lys 
            660                 665                 670         


Pro Asp Glu Arg Asp Arg Thr Leu Ile Val Ser Gly Thr Asn Glu Ala 
        675                 680                 685             


Arg Arg Glu Ile Asn Gln Ile Val Arg Glu Gly Leu Gly Thr Ala Gly 
    690                 695                 700                 


Lys Gly Ile Glu Phe Asp Thr Leu Val Arg Val Asp Thr Thr Gln Ala 
705                 710                 715                 720 


Glu Arg Arg His Ser Lys Asn Tyr Gln Val Gly His Val Ile Gln Pro 
                725                 730                 735     


Glu Arg Asp Tyr Ala Lys Thr Gly Leu Gln Arg Gly Glu Leu Tyr Arg 
            740                 745                 750         


Val Val Glu Thr Gly Pro Gly Asn Arg Leu Thr Val Ile Gly Glu His 
        755                 760                 765             


Asp Gly Gln Arg Ile Gln Phe Ser Pro Met Thr His Thr Lys Ile Ser 
    770                 775                 780                 


Val Tyr Gln Pro Glu Arg Ala Glu Leu Ala Val Gly Asp Thr Ile Arg 
785                 790                 795                 800 


Ile Thr Arg Asn Asp Lys His Leu Asp Leu Ala Asn Gly Asp Arg Met 
                805                 810                 815     


Lys Val Val Ala Val Glu Asp Arg Lys Val Thr Val Thr Asp Gly Lys 
            820                 825                 830         


Arg Asn Val Glu Leu Pro Thr Asp Lys Pro Leu His Val Asp His Ala 
        835                 840                 845             


Tyr Ala Thr Thr Val His Ser Ser Gln Gly Leu Thr Ser Asp Arg Val 
    850                 855                 860                 


Leu Ile Asp Ala His Ala Glu Ser Arg Thr Thr Ala Lys Asp Val Tyr 
865                 870                 875                 880 


Tyr Val Ala Ile Ser Arg Ala Arg Phe Glu Ala Arg Val Phe Thr Asn 
                885                 890                 895     


Asp Arg Gly Lys Leu Pro Ala Ala Ile Ala Arg Glu Asn Ile Lys Ser 
            900                 905                 910         


Ala Ala His Asp Leu Ala Arg Asp Arg Gly Gly Arg Ser Ala Ala Ala 
        915                 920                 925             


Glu Arg Gln Arg Glu Gln Gln Arg Glu Ala Glu Arg Asn Arg Gln Thr 
    930                 935                 940                 


Gln Gln Pro Ala His Asp Arg Gln Lys Ala Ala Arg Glu Ala Glu Arg 
945                 950                 955                 960 


Gly Met Glu Ala Gly Arg Met Thr Gln Phe Glu Gly Phe Thr Asn Leu 
                965                 970                 975     


Tyr Gln Val Ser Lys Thr Leu Arg Phe Glu Leu Ile Pro Gln Gly Lys 
            980                 985                 990         


Thr Leu Lys His Ile Gln Glu Gln  Gly Phe Ile Glu Glu  Asp Lys Ala 
        995                 1000                 1005             


Arg Asn  Asp His Tyr Lys Glu  Leu Lys Pro Ile Ile  Asp Arg Ile 
    1010                 1015                 1020             


Tyr Lys  Thr Tyr Ala Asp Gln  Cys Leu Gln Leu Val  Gln Leu Asp 
    1025                 1030                 1035             


Trp Glu  Asn Leu Ser Ala Ala  Ile Asp Ser Tyr Arg  Lys Glu Lys 
    1040                 1045                 1050             


Thr Glu  Glu Thr Arg Asn Ala  Leu Ile Glu Glu Gln  Ala Thr Tyr 
    1055                 1060                 1065             


Arg Asn  Ala Ile His Asp Tyr  Phe Ile Gly Arg Thr  Asp Asn Leu 
    1070                 1075                 1080             


Thr Asp  Ala Ile Asn Lys Arg  His Ala Glu Ile Tyr  Lys Gly Leu 
    1085                 1090                 1095             


Phe Lys  Ala Glu Leu Phe Asn  Gly Lys Val Leu Lys  Gln Leu Gly 
    1100                 1105                 1110             


Thr Val  Thr Thr Thr Glu His  Glu Asn Ala Leu Leu  Arg Ser Phe 
    1115                 1120                 1125             


Asp Lys  Phe Thr Thr Tyr Phe  Ser Gly Phe Tyr Glu  Asn Arg Lys 
    1130                 1135                 1140             


Asn Val  Phe Ser Ala Glu Asp  Ile Ser Thr Ala Ile  Pro His Arg 
    1145                 1150                 1155             


Ile Val  Gln Asp Asn Phe Pro  Lys Phe Lys Glu Asn  Cys His Ile 
    1160                 1165                 1170             


Phe Thr  Arg Leu Ile Thr Ala  Val Pro Ser Leu Arg  Glu His Phe 
    1175                 1180                 1185             


Glu Asn  Val Lys Lys Ala Ile  Gly Ile Phe Val Ser  Thr Ser Ile 
    1190                 1195                 1200             


Glu Glu  Val Phe Ser Phe Pro  Phe Tyr Asn Gln Leu  Leu Thr Gln 
    1205                 1210                 1215             


Thr Gln  Ile Asp Leu Tyr Asn  Gln Leu Leu Gly Gly  Ile Ser Arg 
    1220                 1225                 1230             


Glu Ala  Gly Thr Glu Lys Ile  Lys Gly Leu Asn Glu  Val Leu Asn 
    1235                 1240                 1245             


Leu Ala  Ile Gln Lys Asn Asp  Glu Thr Ala His Ile  Ile Ala Ser 
    1250                 1255                 1260             


Leu Pro  His Arg Phe Ile Pro  Leu Phe Lys Gln Ile  Leu Ser Asp 
    1265                 1270                 1275             


Arg Asn  Thr Leu Ser Phe Ile  Leu Glu Glu Phe Lys  Ser Asp Glu 
    1280                 1285                 1290             


Glu Val  Ile Gln Ser Phe Cys  Lys Tyr Lys Thr Leu  Leu Arg Asn 
    1295                 1300                 1305             


Glu Asn  Val Leu Glu Thr Ala  Glu Ala Leu Phe Asn  Glu Leu Asn 
    1310                 1315                 1320             


Ser Ile  Asp Leu Thr His Ile  Phe Ile Ser His Lys  Lys Leu Glu 
    1325                 1330                 1335             


Thr Ile  Ser Ser Ala Leu Cys  Asp His Trp Asp Thr  Leu Arg Asn 
    1340                 1345                 1350             


Ala Leu  Tyr Glu Arg Arg Ile  Ser Glu Leu Thr Gly  Lys Ile Thr 
    1355                 1360                 1365             


Lys Ser  Ala Lys Glu Lys Val  Gln Arg Ser Leu Lys  His Glu Asp 
    1370                 1375                 1380             


Ile Asn  Leu Gln Glu Ile Ile  Ser Ala Ala Gly Lys  Glu Leu Ser 
    1385                 1390                 1395             


Glu Ala  Phe Lys Gln Lys Thr  Ser Glu Ile Leu Ser  His Ala His 
    1400                 1405                 1410             


Ala Ala  Leu Asp Gln Pro Leu  Pro Thr Thr Leu Lys  Lys Gln Glu 
    1415                 1420                 1425             


Glu Lys  Glu Ile Leu Lys Ser  Gln Leu Asp Ser Leu  Leu Gly Leu 
    1430                 1435                 1440             


Tyr His  Leu Leu Asp Trp Phe  Ala Val Asp Glu Ser  Asn Glu Val 
    1445                 1450                 1455             


Asp Pro  Glu Phe Ser Ala Arg  Leu Thr Gly Ile Lys  Leu Glu Met 
    1460                 1465                 1470             


Glu Pro  Ser Leu Ser Phe Tyr  Asn Lys Ala Arg Asn  Tyr Ala Thr 
    1475                 1480                 1485             


Lys Lys  Pro Tyr Ser Val Glu  Lys Phe Lys Leu Asn  Phe Gln Met 
    1490                 1495                 1500             


Pro Thr  Leu Ala Ser Gly Trp  Asp Val Asn Lys Glu  Lys Asn Asn 
    1505                 1510                 1515             


Gly Ala  Ile Leu Phe Val Lys  Asn Gly Leu Tyr Tyr  Leu Gly Ile 
    1520                 1525                 1530             


Met Pro  Lys Gln Lys Gly Arg  Tyr Lys Ala Leu Ser  Phe Glu Pro 
    1535                 1540                 1545             


Thr Glu  Lys Thr Ser Glu Gly  Phe Asp Lys Met Tyr  Tyr Asp Tyr 
    1550                 1555                 1560             


Phe Pro  Asp Ala Ala Lys Met  Ile Pro Lys Cys Ser  Thr Gln Leu 
    1565                 1570                 1575             


Lys Ala  Val Thr Ala His Phe  Gln Thr His Thr Thr  Pro Ile Leu 
    1580                 1585                 1590             


Leu Ser  Asn Asn Phe Ile Glu  Pro Leu Glu Ile Thr  Lys Glu Ile 
    1595                 1600                 1605             


Tyr Asp  Leu Asn Asn Pro Glu  Lys Glu Pro Lys Lys  Phe Gln Thr 
    1610                 1615                 1620             


Ala Tyr  Ala Lys Lys Thr Gly  Asp Gln Lys Gly Tyr  Arg Glu Ala 
    1625                 1630                 1635             


Leu Cys  Lys Trp Ile Asp Phe  Thr Arg Asp Phe Leu  Ser Lys Tyr 
    1640                 1645                 1650             


Thr Lys  Thr Thr Ser Ile Asp  Leu Ser Ser Leu Arg  Pro Ser Ser 
    1655                 1660                 1665             


Gln Tyr  Lys Asp Leu Gly Glu  Tyr Tyr Ala Glu Leu  Asn Pro Leu 
    1670                 1675                 1680             


Leu Tyr  His Ile Ser Phe Gln  Arg Ile Ala Glu Lys  Glu Ile Met 
    1685                 1690                 1695             


Asp Ala  Val Glu Thr Gly Lys  Leu Tyr Leu Phe Gln  Ile Tyr Asn 
    1700                 1705                 1710             


Lys Asp  Phe Ala Lys Gly His  His Gly Lys Pro Asn  Leu His Thr 
    1715                 1720                 1725             


Leu Tyr  Trp Thr Gly Leu Phe  Ser Pro Glu Asn Leu  Ala Lys Thr 
    1730                 1735                 1740             


Ser Ile  Lys Leu Asn Gly Gln  Ala Glu Leu Phe Tyr  Arg Pro Lys 
    1745                 1750                 1755             


Ser Arg  Met Lys Arg Met Ala  His Arg Leu Gly Glu  Lys Met Leu 
    1760                 1765                 1770             


Asn Lys  Lys Leu Lys Asp Gln  Lys Thr Pro Ile Pro  Asp Thr Leu 
    1775                 1780                 1785             


Tyr Gln  Glu Leu Tyr Asp Tyr  Val Asn His Arg Leu  Ser His Asp 
    1790                 1795                 1800             


Leu Ser  Asp Glu Ala Arg Ala  Leu Leu Pro Asn Val  Ile Thr Lys 
    1805                 1810                 1815             


Glu Val  Ser His Glu Ile Ile  Lys Asp Arg Arg Phe  Thr Ser Asp 
    1820                 1825                 1830             


Lys Phe  Phe Phe His Val Pro  Ile Thr Leu Asn Tyr  Gln Ala Ala 
    1835                 1840                 1845             


Asn Ser  Pro Ser Lys Phe Asn  Gln Arg Val Asn Ala  Tyr Leu Lys 
    1850                 1855                 1860             


Glu His  Pro Glu Thr Pro Ile  Ile Gly Ile Asp Arg  Gly Glu Arg 
    1865                 1870                 1875             


Asn Leu  Ile Tyr Ile Thr Val  Ile Asp Ser Thr Gly  Lys Ile Leu 
    1880                 1885                 1890             


Glu Gln  Arg Ser Leu Asn Thr  Ile Gln Gln Phe Asp  Tyr Gln Lys 
    1895                 1900                 1905             


Lys Leu  Asp Asn Arg Glu Lys  Glu Arg Val Ala Ala  Arg Gln Ala 
    1910                 1915                 1920             


Trp Ser  Val Val Gly Thr Ile  Lys Asp Leu Lys Gln  Gly Tyr Leu 
    1925                 1930                 1935             


Ser Gln  Val Ile His Glu Ile  Val Asp Leu Met Ile  His Tyr Gln 
    1940                 1945                 1950             


Ala Val  Val Val Leu Glu Asn  Leu Asn Phe Gly Phe  Lys Ser Lys 
    1955                 1960                 1965             


Arg Thr  Gly Ile Ala Glu Lys  Ala Val Tyr Gln Gln  Phe Glu Lys 
    1970                 1975                 1980             


Met Leu  Ile Asp Lys Leu Asn  Cys Leu Val Leu Lys  Asp Tyr Pro 
    1985                 1990                 1995             


Ala Glu  Lys Val Gly Gly Val  Leu Asn Pro Tyr Gln  Leu Thr Asp 
    2000                 2005                 2010             


Gln Phe  Thr Ser Phe Ala Lys  Met Gly Thr Gln Ser  Gly Phe Leu 
    2015                 2020                 2025             


Phe Tyr  Val Pro Ala Pro Tyr  Thr Ser Lys Ile Asp  Pro Leu Thr 
    2030                 2035                 2040             


Gly Phe  Val Asp Pro Phe Val  Trp Lys Thr Ile Lys  Asn His Glu 
    2045                 2050                 2055             


Ser Arg  Lys His Phe Leu Glu  Gly Phe Asp Phe Leu  His Tyr Asp 
    2060                 2065                 2070             


Val Lys  Thr Gly Asp Phe Ile  Leu His Phe Lys Met  Asn Arg Asn 
    2075                 2080                 2085             


Leu Ser  Phe Gln Arg Gly Leu  Pro Gly Phe Met Pro  Ala Trp Asp 
    2090                 2095                 2100             


Ile Val  Phe Glu Lys Asn Glu  Thr Gln Phe Asp Ala  Lys Gly Thr 
    2105                 2110                 2115             


Pro Phe  Ile Ala Gly Lys Arg  Ile Val Pro Val Ile  Glu Asn His 
    2120                 2125                 2130             


Arg Phe  Thr Gly Arg Tyr Arg  Asp Leu Tyr Pro Ala  Asn Glu Leu 
    2135                 2140                 2145             


Ile Ala  Leu Leu Glu Glu Lys  Gly Ile Val Phe Arg  Asp Gly Ser 
    2150                 2155                 2160             


Asn Ile  Leu Pro Lys Leu Leu  Glu Asn Asp Asp Ser  His Ala Ile 
    2165                 2170                 2175             


Asp Thr  Met Val Ala Leu Ile  Arg Ser Val Leu Gln  Met Arg Asn 
    2180                 2185                 2190             


Ser Asn  Ala Ala Thr Gly Glu  Asp Tyr Ile Asn Ser  Pro Val Arg 
    2195                 2200                 2205             


Asp Leu  Asn Gly Val Cys Phe  Asp Ser Arg Phe Gln  Asn Pro Glu 
    2210                 2215                 2220             


Trp Pro  Met Asp Ala Asp Ala  Asn Gly Ala Tyr His  Ile Ala Leu 
    2225                 2230                 2235             


Lys Gly  Gln Leu Leu Leu Asn  His Leu Lys Glu Ser  Lys Asp Leu 
    2240                 2245                 2250             


Lys Leu  Gln Asn Gly Ile Ser  Asn Gln Asp Trp Leu  Ala Tyr Ile 
    2255                 2260                 2265             


Gln Glu  Leu Arg Asn Lys Arg  Pro Ala Ala Thr Lys  Lys Ala Gly 
    2270                 2275                 2280             


Gln Ala  Lys Lys Lys Lys Gly  Ser Tyr Pro Tyr Asp  Val Pro Asp 
    2285                 2290                 2295             


Tyr Ala  Tyr Pro Tyr Asp Val  Pro Asp Tyr Ala Tyr  Pro Tyr Asp 
    2300                 2305                 2310             


Val Pro  Asp Tyr Ala 
    2315             


<210>  2
<211>  6957
<212>  DNA
<213>  Secuencia Artificial

<220>
<223>  TrwC:Cas12 - DNA

<400>  2
atgctcagtc acatggtatt gacccgacag gacatagggc gggctgcaag ctattacgag       60

gacggcgcgg acgattacta tgccaaggat ggcgatgcat cagaatggca ggggaagggt      120

gcggaagaac tcggcttgag tggggaggtg gacagcaaac ggttccgcga gctgctggcc      180

gggaatatcg gcgagggcca ccgcatcatg cgctcggcta ctcgacagga cagcaaagag      240

cgcatcggcc ttgacctcac attcagcgcg cccaagagcg tatcgcttca agcccttgtt      300

gcgggcgatg ctgaaatcat caaagcgcac gaccgcgcag ttgctcgcac cttggagcag      360

gccgaagccc gcgcacaggc ccgccaaaag atacaaggca agacccgcat cgagacgacc      420

gggaatttgg tcatcggtaa atttcgccat gagacaagcc gcgagcgcga tccgcagcta      480

catacgcacg cggtcattct caacatgacc aaacgtagtg acggccaatg gcgggcgttg      540

aaaaacgacg agattgtgaa ggctacgcgc taccttggag ccgtctataa cgcggagctg      600

gcccacgaac tgcaaaagct cggttatcag cttcggtacg gcaaagacgg taatttcgac      660

ctagcacaca tcgaccgaca gcaaatcgag ggattcagca aacgcacgga acagattgcc      720

gaatggtatg ccgcccgtgg tcttgaccct aatagcgtat ctttggaaca gaaacaggcg      780

gccaaggttt tgagccgtgc taagaaaact tcggtagacc gggaggcatt gcgggctgaa      840

tggcaagcca cggcaaaaga actcggcata gatttcagcc gccgggaatg gtcaggccgg      900

gagaagggcg gttcagaaaa acaggcccat tcattcatgc cttccgacga ggcggccaag      960

cgggccgttc gctatgccat caaccatctg acagagcggc aatcagtcat ggacgagcgc     1020

gagcttgtcg atacggccat gaagcatgcc gtaggcgcag cccggttaga ggacattcag     1080

aaggagcttt tgcgtcagac cgaaaccggc tacctcattc gagaagcccc gcgctatcgc     1140

ccaggcgggc agaccgggcc aacggatgaa cccggcaaga cccgcgccga gtgggttgca     1200

gagctggccg ctaagggcat gaagcaaggc gcagcacgcg agcgcgtcga taacgccatc     1260

aagaccggcg ggcttgtacc gatagaaccg cgttacacca cacaaacggc gcttgagcgc     1320

gagaaacgca tcttgcagat cgaacgtgac ggacgcggcg cagttgcgcc ggtcattgcc     1380

gccgaggccg cccgcgagcg gcttgccagc acgaatctaa accaaggcca gcgcgaagcg     1440

gccgagttga tcgtttcggc tgccaaccgg gttgtcggtg tgcagggttt tgcgggaacc     1500

ggcaagagcc acatgcttga taccgccaag caaatgatcg agggcgaggg ttatcacgtt     1560

cgcgcactgg ccgcctatgg cagtcaggtc aaagcattgc gcgagttgaa cgttgaggcg     1620

aacaccttgg catcgtttct ccgggccaag gataagaaca tcgacagccg caccgtgctt     1680

gtcattgacg aggcgggcgt tgttccaacg cgcttgatgg agcaaacgct aaagctggca     1740

gagaaggccg gggcgcgtgt tgtgcttatg ggtgataccg cgcaaaccaa ggccatcgag     1800

gcgggccgac ctttcgacca gctacaggcc gccgggatgc agaccgcgca catgcgggag     1860

attcagcgcc agaagaaccc cgagctgaaa atcgccgtcg agctggccgc agcaggcaag     1920

gcaagcagct cgctagagcg catcaaagac gtaacggaaa tcaaaaacca tcatgaacgt     1980

agggctgccg tggccgaggc atatatcgcg ctcaagccag acgagcgcga ccggacgctg     2040

attgtatcgg gcaccaacga ggcacgccgt gaaatcaatc agatcgtgcg cgagggcttg     2100

ggaacggcag ggaagggcat cgagttcgat acccttgtgc gcgtggacac gacgcaggcc     2160

gagcgccgcc actcgaaaaa ctaccaagtc gggcatgtca tccagccaga gcgggactat     2220

gccaaaaccg gcttgcagcg tggcgagttg taccgcgtag tcgaaaccgg gccgggaaac     2280

cggctaaccg ttatcggcga gcatgacggc caacgaatcc agttcagccc catgacacac     2340

acaaagattt cggtctatca gccggagcgg gccgagctgg ccgttggcga cacaatacgc     2400

atcacgcgaa acgacaaaca ccttgacctg gctaacggcg accgaatgaa agttgtcgcg     2460

gttgaagatc ggaaagtcac cgtgacggac ggaaagcgaa acgtcgagct gcccaccgat     2520

aagccgctgc acgtcgatca tgcctatgcg acgactgttc acagcagcca agggctaacg     2580

tctgatcggg tgttgatcga cgcccatgcc gagagccgta ccacggccaa ggacgtttac     2640

tatgtggcga tttcccgcgc ccgttttgag gcacgggttt tcacgaacga ccggggcaag     2700

ctgcccgccg ccattgcgcg ggaaaacatc aagagcgccg cgcatgacct cgcccgcgac     2760

cggggaggcc gtagcgccgc agcagagcgc cagagggagc aacagcgcga agcagagcgc     2820

aaccggcaga cgcaacagcc cgcgcatgat cggcagaagg ccgccagaga agccgagcgc     2880

ggcatggagg ccggaaggat gacacagttc gagggcttta ccaacctgta tcaggtgagc     2940

aagacactgc ggtttgagct gatcccacag ggcaagaccc tgaagcacat ccaggagcag     3000

ggcttcatcg aggaggacaa ggcccgcaat gatcactaca aggagctgaa gcccatcatc     3060

gatcggatct acaagaccta tgccgaccag tgcctgcagc tggtgcagct ggattgggag     3120

aacctgagcg ccgccatcga ctcctataga aaggagaaaa ccgaggagac aaggaacgcc     3180

ctgatcgagg agcaggccac atatcgcaat gccatccacg actacttcat cggccggaca     3240

gacaacctga ccgatgccat caataagaga cacgccgaga tctacaaggg cctgttcaag     3300

gccgagctgt ttaatggcaa ggtgctgaag cagctgggca ccgtgaccac aaccgagcac     3360

gagaacgccc tgctgcggag cttcgacaag tttacaacct acttctccgg cttttatgag     3420

aacaggaaga acgtgttcag cgccgaggat atcagcacag ccatcccaca ccgcatcgtg     3480

caggacaact tccccaagtt taaggagaat tgtcacatct tcacacgcct gatcaccgcc     3540

gtgcccagcc tgcgggagca ctttgagaac gtgaagaagg ccatcggcat cttcgtgagc     3600

acctccatcg aggaggtgtt ttccttccct ttttataacc agctgctgac acagacccag     3660

atcgacctgt ataaccagct gctgggagga atctctcggg aggcaggcac cgagaagatc     3720

aagggcctga acgaggtgct gaatctggcc atccagaaga atgatgagac agcccacatc     3780

atcgcctccc tgccacacag attcatcccc ctgtttaagc agatcctgtc cgataggaac     3840

accctgtctt tcatcctgga ggagtttaag agcgacgagg aagtgatcca gtccttctgc     3900

aagtacaaga cactgctgag aaacgagaac gtgctggaga cagccgaggc cctgtttaac     3960

gagctgaaca gcatcgacct gacacacatc ttcatcagcc acaagaagct ggagacaatc     4020

agcagcgccc tgtgcgacca ctgggataca ctgaggaatg ccctgtatga gcggagaatc     4080

tccgagctga caggcaagat caccaagtct gccaaggaga aggtgcagcg cagcctgaag     4140

cacgaggata tcaacctgca ggagatcatc tctgccgcag gcaaggagct gagcgaggcc     4200

ttcaagcaga aaaccagcga gatcctgtcc cacgcacacg ccgccctgga tcagccactg     4260

cctacaaccc tgaagaagca ggaggagaag gagatcctga agtctcagct ggacagcctg     4320

ctgggcctgt accacctgct ggactggttt gccgtggatg agtccaacga ggtggacccc     4380

gagttctctg cccggctgac cggcatcaag ctggagatgg agccttctct gagcttctac     4440

aacaaggcca gaaattatgc caccaagaag ccctactccg tggagaagtt caagctgaac     4500

tttcagatgc ctacactggc ctctggctgg gacgtgaata aggagaagaa caatggcgcc     4560

atcctgtttg tgaagaacgg cctgtactat ctgggcatca tgccaaagca gaagggcagg     4620

tataaggccc tgagcttcga gcccacagag aaaaccagcg agggctttga taagatgtac     4680

tatgactact tccctgatgc cgccaagatg atcccaaagt gcagcaccca gctgaaggcc     4740

gtgacagccc actttcagac ccacacaacc cccatcctgc tgtccaacaa tttcatcgag     4800

cctctggaga tcacaaagga gatctacgac ctgaacaatc ctgagaagga gccaaagaag     4860

tttcagacag cctacgccaa gaaaaccggc gaccagaagg gctacagaga ggccctgtgc     4920

aagtggatcg acttcacaag ggattttctg tccaagtata ccaagacaac ctctatcgat     4980

ctgtctagcc tgcggccatc ctctcagtat aaggacctgg gcgagtacta tgccgagctg     5040

aatcccctgc tgtaccacat cagcttccag agaatcgccg agaaggagat catggatgcc     5100

gtggagacag gcaagctgta cctgttccag atctataaca aggactttgc caagggccac     5160

cacggcaagc ctaatctgca cacactgtat tggaccggcc tgttttctcc agagaacctg     5220

gccaagacaa gcatcaagct gaatggccag gccgagctgt tctaccgccc taagtccagg     5280

atgaagagga tggcacaccg gctgggagag aagatgctga acaagaagct gaaggatcag     5340

aaaaccccaa tccccgacac cctgtaccag gagctgtacg actatgtgaa tcacagactg     5400

tcccacgacc tgtctgatga ggccagggcc ctgctgccca acgtgatcac caaggaggtg     5460

tctcacgaga tcatcaagga taggcgcttt accagcgaca agttcttttt ccacgtgcct     5520

atcacactga actatcaggc cgccaattcc ccatctaagt tcaaccagag ggtgaatgcc     5580

tacctgaagg agcaccccga gacacctatc atcggcatcg atcggggcga gagaaacctg     5640

atctatatca cagtgatcga ctccaccggc aagatcctgg agcagcggag cctgaacacc     5700

atccagcagt ttgattacca gaagaagctg gacaacaggg agaaggagag ggtggcagca     5760

aggcaggcct ggtctgtggt gggcacaatc aaggatctga agcagggcta tctgagccag     5820

gtcatccacg agatcgtgga cctgatgatc cactaccagg ccgtggtggt gctggagaac     5880

ctgaatttcg gctttaagag caagaggacc ggcatcgccg agaaggccgt gtaccagcag     5940

ttcgagaaga tgctgatcga taagctgaat tgcctggtgc tgaaggacta tccagcagag     6000

aaagtgggag gcgtgctgaa cccataccag ctgacagacc agttcacctc ctttgccaag     6060

atgggcaccc agtctggctt cctgttttac gtgcctgccc catatacatc taagatcgat     6120

cccctgaccg gcttcgtgga ccccttcgtg tggaaaacca tcaagaatca cgagagccgc     6180

aagcacttcc tggagggctt cgactttctg cactacgacg tgaaaaccgg cgacttcatc     6240

ctgcacttta agatgaacag aaatctgtcc ttccagaggg gcctgcccgg ctttatgcct     6300

gcatgggata tcgtgttcga gaagaacgag acacagtttg acgccaaggg cacccctttc     6360

atcgccggca agagaatcgt gccagtgatc gagaatcaca gattcaccgg cagataccgg     6420

gacctgtatc ctgccaacga gctgatcgcc ctgctggagg agaagggcat cgtgttcagg     6480

gatggctcca acatcctgcc aaagctgctg gagaatgacg attctcacgc catcgacacc     6540

atggtggccc tgatccgcag cgtgctgcag atgcggaact ccaatgccgc cacaggcgag     6600

gactatatca acagccccgt gcgcgatctg aatggcgtgt gcttcgactc ccggtttcag     6660

aacccagagt ggcccatgga cgccgatgcc aatggcgcct accacatcgc cctgaagggc     6720

cagctgctgc tgaatcacct gaaggagagc aaggatctga agctgcagaa cggcatctcc     6780

aatcaggact ggctggccta catccaggag ctgcgcaaca aaaggccggc ggccacgaaa     6840

aaggccggcc aggcaaaaaa gaaaaaggga tcctacccat acgatgttcc agattacgct     6900

tatccctacg acgtgcctga ttatgcatac ccatatgatg tccccgacta tgcctaa        6957


<210>  3
<211>  41
<212>  DNA
<213>  Secuencia Artificial

<220>
<223>  pY010_TrwC_Forward

<400>  3
ttaagcttgg taccgccacc atgctcagtc acatggtatt g                           41


<210>  4
<211>  35
<212>  DNA
<213>  Secuencia Artificial

<220>
<223>  Cpf1_TrwC_R

<400>  4
aagccctcga actgtgtcat ccttccggcc tccat                                  35


<210>  5
<211>  15
<212>  DNA
<213>  Secuencia Artificial

<220>
<223>  Cpf1_1_F

<400>  5
ggtggcggta ccaag                                                        15


<210>  6
<211>  17
<212>  DNA
<213>  Secuencia Artificial

<220>
<223>  pY010_929_R

<400>  6
atgacacagt tcgaggg                                                      17


<210>  7
<211>  50
<212>  DNA
<213>  Secuencia Artificial

<220>
<223>  Cpf1_pBBR6_F

<400>  7
tgagcggata acaatttcac acaggaaaca gctatgacac agttcgaggg                  50


<210>  8
<211>  47
<212>  DNA
<213>  Secuencia Artificial

<220>
<223>  pLG15_Cpf1_R

<400>  8
tttaacgcga attttaacaa aatattaacg cggcatagtc ggggaca                     47


<210>  9
<211>  24
<212>  DNA
<213>  Secuencia Artificial

<220>
<223>  pBBR6_1440_F

<400>  9
gcgttaatat tttgttaaaa ttcg                                              24


<210>  10
<211>  19
<212>  DNA
<213>  Secuencia Artificial

<220>
<223>  pBBR6_1074_R

<400>  10
agctgtttcc tgtgtgaaa                                                    19


<210>  11
<211>  107
<212>  DNA
<213>  Secuencia Artificial

<220>
<223>  lacZ-gRNA

<400>  11
gaattcgtca aaagaccttt ttaatttcta ctcttgtaga tccgaccgca cgccgcatcc       60

agcgctgtca aaagaccttt ttaatttcta ctcttgtaga taagctt                    107


<210>  12
<211>  105
<212>  DNA
<213>  Secuencia Artificial

<220>
<223>  sacB-gRNA

<400>  12
gaattcgtca aaagaccttt ttaatttcta ctcttgtaga tggacagctg gccattacaa       60

aacggtcaaa agaccttttt aatttctact cttgtagata agctt                      105


<210>  13
<211>  24
<212>  DNA
<213>  Secuencia Artificial

<220>
<223>  tetR_F

<400>  13
ttaagaccca ctttcacatt taag                                              24


<210>  14
<211>  22
<212>  DNA
<213>  Secuencia Artificial

<220>
<223>  ptet_R

<400>  14
ttttgcctcc taactaggtc at                                                22


<210>  15
<211>  62
<212>  DNA
<213>  Secuencia Artificial

<220>
<223>  ptet_TrwC_F

<400>  15
tatccggagg catatcaaat gacctagtta ggaggcaaaa atgctcagtc acatggtatt       60

ga                                                                      62


<210>  16
<211>  58
<212>  DNA
<213>  Secuencia Artificial

<220>
<223>  TrwC_Cpf1_R

<400>  16
tcacctgata caggttggta aagccctcga actgtgtcat tgaccttccg gcctccat         58


<210>  17
<211>  19
<212>  DNA
<213>  Secuencia Artificial

<220>
<223>  Cpf1_F

<400>  17
atgacacagt tcgagggct                                                    19


<210>  18
<211>  16
<212>  DNA
<213>  Secuencia Artificial

<220>
<223>  pLG14_R

<400>  18
gcgtagcacc aggcgt                                                       16


<210>  19
<211>  20
<212>  DNA
<213>  Secuencia Artificial

<220>
<223>  pLG14_F

<400>  19
accaataggc cgactgcgat                                                   20


<210>  20
<211>  62
<212>  DNA
<213>  Secuencia Artificial

<220>
<223>  pLG14_2_R

<400>  20
cggattagaa aaacaactta aatgtgaaag tgggtcttaa ttaggtggcg gtacttgggt       60

cg                                                                      62


<210>  21
<211>  24
<212>  DNA
<213>  Secuencia Artificial

<220>
<223>  U6_crRNA_F

<400>  21
gagggcctat ttcccatgat tcct                                              24


<210>  22
<211>  66
<212>  DNA
<213>  Secuencia Artificial

<220>
<223>  DNMT1_crRNA3_R

<400>  22
gagtaacaga catggaccat cagatctaca agagtagaaa ttacggtgtt tcgtcctttc       60

cacaag                                                                  66


<210>  23
<211>  20
<212>  DNA
<213>  Secuencia Artificial

<220>
<223>  DNMT1_F

<400>  23
ctgggactca ggcgggtcac                                                   20


<210>  24
<211>  25
<212>  DNA
<213>  Secuencia Artificial

<220>
<223>  DNMT1_R

<400>  24
cctcacacaa cagcttcatg tcagc                                             25


<210>  25
<211>  18
<212>  DNA
<213>  Secuencia Artificial

<220>
<223>  SacB_F

<400>  25
ctaccgcact gctggcag                                                     18


<210>  26
<211>  20
<212>  DNA
<213>  Secuencia Artificial

<220>
<223>  SacB_R

<400>  26
gatgctgtct ttgacaacag                                                   20


