                         SEQUENCE LISTING

<110>  CellResearch Corporation Pte Ltd
       Singapore Health Services Pte Ltd.
 
<120>  A Method Of Generating A Mammalian Stem Cell Carrying a 
       Transgene, a Mammalian Stem Cell Generated By The Method and 
       Pharmaceuticals Uses Of The Mammalian Stem Cell

<130>  CRC15610PCT

<150>  US62/267,056
<151>  2015-12-14

<160>  32    

<170>  PatentIn version 3.5

<210>  1
<211>  2351
<212>  PRT
<213>  Homo sapiens

<400>  1

Met Gln Ile Glu Leu Ser Thr Cys Phe Phe Leu Cys Leu Leu Arg Phe 
1               5                   10                  15      


Cys Phe Ser Ala Thr Arg Arg Tyr Tyr Leu Gly Ala Val Glu Leu Ser 
            20                  25                  30          


Trp Asp Tyr Met Gln Ser Asp Leu Gly Glu Leu Pro Val Asp Ala Arg 
        35                  40                  45              


Phe Pro Pro Arg Val Pro Lys Ser Phe Pro Phe Asn Thr Ser Val Val 
    50                  55                  60                  


Tyr Lys Lys Thr Leu Phe Val Glu Phe Thr Asp His Leu Phe Asn Ile 
65                  70                  75                  80  


Ala Lys Pro Arg Pro Pro Trp Met Gly Leu Leu Gly Pro Thr Ile Gln 
                85                  90                  95      


Ala Glu Val Tyr Asp Thr Val Val Ile Thr Leu Lys Asn Met Ala Ser 
            100                 105                 110         


His Pro Val Ser Leu His Ala Val Gly Val Ser Tyr Trp Lys Ala Ser 
        115                 120                 125             


Glu Gly Ala Glu Tyr Asp Asp Gln Thr Ser Gln Arg Glu Lys Glu Asp 
    130                 135                 140                 


Asp Lys Val Phe Pro Gly Gly Ser His Thr Tyr Val Trp Gln Val Leu 
145                 150                 155                 160 


Lys Glu Asn Gly Pro Met Ala Ser Asp Pro Leu Cys Leu Thr Tyr Ser 
                165                 170                 175     


Tyr Leu Ser His Val Asp Leu Val Lys Asp Leu Asn Ser Gly Leu Ile 
            180                 185                 190         


Gly Ala Leu Leu Val Cys Arg Glu Gly Ser Leu Ala Lys Glu Lys Thr 
        195                 200                 205             


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


Lys Ser Trp His Ser Glu Thr Lys Asn Ser Leu Met Gln Asp Arg Asp 
225                 230                 235                 240 


Ala Ala Ser Ala Arg Ala Trp Pro Lys Met His Thr Val Asn Gly Tyr 
                245                 250                 255     


Val Asn Arg Ser Leu Pro Gly Leu Ile Gly Cys His Arg Lys Ser Val 
            260                 265                 270         


Tyr Trp His Val Ile Gly Met Gly Thr Thr Pro Glu Val His Ser Ile 
        275                 280                 285             


Phe Leu Glu Gly His Thr Phe Leu Val Arg Asn His Arg Gln Ala Ser 
    290                 295                 300                 


Leu Glu Ile Ser Pro Ile Thr Phe Leu Thr Ala Gln Thr Leu Leu Met 
305                 310                 315                 320 


Asp Leu Gly Gln Phe Leu Leu Phe Cys His Ile Ser Ser His Gln His 
                325                 330                 335     


Asp Gly Met Glu Ala Tyr Val Lys Val Asp Ser Cys Pro Glu Glu Pro 
            340                 345                 350         


Gln Leu Arg Met Lys Asn Asn Glu Glu Ala Glu Asp Tyr Asp Asp Asp 
        355                 360                 365             


Leu Thr Asp Ser Glu Met Asp Val Val Arg Phe Asp Asp Asp Asn Ser 
    370                 375                 380                 


Pro Ser Phe Ile Gln Ile Arg Ser Val Ala Lys Lys His Pro Lys Thr 
385                 390                 395                 400 


Trp Val His Tyr Ile Ala Ala Glu Glu Glu Asp Trp Asp Tyr Ala Pro 
                405                 410                 415     


Leu Val Leu Ala Pro Asp Asp Arg Ser Tyr Lys Ser Gln Tyr Leu Asn 
            420                 425                 430         


Asn Gly Pro Gln Arg Ile Gly Arg Lys Tyr Lys Lys Val Arg Phe Met 
        435                 440                 445             


Ala Tyr Thr Asp Glu Thr Phe Lys Thr Arg Glu Ala Ile Gln His Glu 
    450                 455                 460                 


Ser Gly Ile Leu Gly Pro Leu Leu Tyr Gly Glu Val Gly Asp Thr Leu 
465                 470                 475                 480 


Leu Ile Ile Phe Lys Asn Gln Ala Ser Arg Pro Tyr Asn Ile Tyr Pro 
                485                 490                 495     


His Gly Ile Thr Asp Val Arg Pro Leu Tyr Ser Arg Arg Leu Pro Lys 
            500                 505                 510         


Gly Val Lys His Leu Lys Asp Phe Pro Ile Leu Pro Gly Glu Ile Phe 
        515                 520                 525             


Lys Tyr Lys Trp Thr Val Thr Val Glu Asp Gly Pro Thr Lys Ser Asp 
    530                 535                 540                 


Pro Arg Cys Leu Thr Arg Tyr Tyr Ser Ser Phe Val Asn Met Glu Arg 
545                 550                 555                 560 


Asp Leu Ala Ser Gly Leu Ile Gly Pro Leu Leu Ile Cys Tyr Lys Glu 
                565                 570                 575     


Ser Val Asp Gln Arg Gly Asn Gln Ile Met Ser Asp Lys Arg Asn Val 
            580                 585                 590         


Ile Leu Phe Ser Val Phe Asp Glu Asn Arg Ser Trp Tyr Leu Thr Glu 
        595                 600                 605             


Asn Ile Gln Arg Phe Leu Pro Asn Pro Ala Gly Val Gln Leu Glu Asp 
    610                 615                 620                 


Pro Glu Phe Gln Ala Ser Asn Ile Met His Ser Ile Asn Gly Tyr Val 
625                 630                 635                 640 


Phe Asp Ser Leu Gln Leu Ser Val Cys Leu His Glu Val Ala Tyr Trp 
                645                 650                 655     


Tyr Ile Leu Ser Ile Gly Ala Gln Thr Asp Phe Leu Ser Val Phe Phe 
            660                 665                 670         


Ser Gly Tyr Thr Phe Lys His Lys Met Val Tyr Glu Asp Thr Leu Thr 
        675                 680                 685             


Leu Phe Pro Phe Ser Gly Glu Thr Val Phe Met Ser Met Glu Asn Pro 
    690                 695                 700                 


Gly Leu Trp Ile Leu Gly Cys His Asn Ser Asp Phe Arg Asn Arg Gly 
705                 710                 715                 720 


Met Thr Ala Leu Leu Lys Val Ser Ser Cys Asp Lys Asn Thr Gly Asp 
                725                 730                 735     


Tyr Tyr Glu Asp Ser Tyr Glu Asp Ile Ser Ala Tyr Leu Leu Ser Lys 
            740                 745                 750         


Asn Asn Ala Ile Glu Pro Arg Ser Phe Ser Gln Asn Ser Arg His Pro 
        755                 760                 765             


Ser Thr Arg Gln Lys Gln Phe Asn Ala Thr Thr Ile Pro Glu Asn Asp 
    770                 775                 780                 


Ile Glu Lys Thr Asp Pro Trp Phe Ala His Arg Thr Pro Met Pro Lys 
785                 790                 795                 800 


Ile Gln Asn Val Ser Ser Ser Asp Leu Leu Met Leu Leu Arg Gln Ser 
                805                 810                 815     


Pro Thr Pro His Gly Leu Ser Leu Ser Asp Leu Gln Glu Ala Lys Tyr 
            820                 825                 830         


Glu Thr Phe Ser Asp Asp Pro Ser Pro Gly Ala Ile Asp Ser Asn Asn 
        835                 840                 845             


Ser Leu Ser Glu Met Thr His Phe Arg Pro Gln Leu His His Ser Gly 
    850                 855                 860                 


Asp Met Val Phe Thr Pro Glu Ser Gly Leu Gln Leu Arg Leu Asn Glu 
865                 870                 875                 880 


Lys Leu Gly Thr Thr Ala Ala Thr Glu Leu Lys Lys Leu Asp Phe Lys 
                885                 890                 895     


Val Ser Ser Thr Ser Asn Asn Leu Ile Ser Thr Ile Pro Ser Asp Asn 
            900                 905                 910         


Leu Ala Ala Gly Thr Asp Asn Thr Ser Ser Leu Gly Pro Pro Ser Met 
        915                 920                 925             


Pro Val His Tyr Asp Ser Gln Leu Asp Thr Thr Leu Phe Gly Lys Lys 
    930                 935                 940                 


Ser Ser Pro Leu Thr Glu Ser Gly Gly Pro Leu Ser Leu Ser Glu Glu 
945                 950                 955                 960 


Asn Asn Asp Ser Lys Leu Leu Glu Ser Gly Leu Met Asn Ser Gln Glu 
                965                 970                 975     


Ser Ser Trp Gly Lys Asn Val Ser Ser Thr Glu Ser Gly Arg Leu Phe 
            980                 985                 990         


Lys Gly Lys Arg Ala His Gly Pro  Ala Leu Leu Thr Lys  Asp Asn Ala 
        995                 1000                 1005             


Leu Phe  Lys Val Ser Ile Ser  Leu Leu Lys Thr Asn  Lys Thr Ser 
    1010                 1015                 1020             


Asn Asn  Ser Ala Thr Asn Arg  Lys Thr His Ile Asp  Gly Pro Ser 
    1025                 1030                 1035             


Leu Leu  Ile Glu Asn Ser Pro  Ser Val Trp Gln Asn  Ile Leu Glu 
    1040                 1045                 1050             


Ser Asp  Thr Glu Phe Lys Lys  Val Thr Pro Leu Ile  His Asp Arg 
    1055                 1060                 1065             


Met Leu  Met Asp Lys Asn Ala  Thr Ala Leu Arg Leu  Asn His Met 
    1070                 1075                 1080             


Ser Asn  Lys Thr Thr Ser Ser  Lys Asn Met Glu Met  Val Gln Gln 
    1085                 1090                 1095             


Lys Lys  Glu Gly Pro Ile Pro  Pro Asp Ala Gln Asn  Pro Asp Met 
    1100                 1105                 1110             


Ser Phe  Phe Lys Met Leu Phe  Leu Pro Glu Ser Ala  Arg Trp Ile 
    1115                 1120                 1125             


Gln Arg  Thr His Gly Lys Asn  Ser Leu Asn Ser Gly  Gln Gly Pro 
    1130                 1135                 1140             


Ser Pro  Lys Gln Leu Val Ser  Leu Gly Pro Glu Lys  Ser Val Glu 
    1145                 1150                 1155             


Gly Gln  Asn Phe Leu Ser Glu  Lys Asn Lys Val Val  Val Gly Lys 
    1160                 1165                 1170             


Gly Glu  Phe Thr Lys Asp Val  Gly Leu Lys Glu Met  Val Phe Pro 
    1175                 1180                 1185             


Ser Ser  Arg Asn Leu Phe Leu  Thr Asn Leu Asp Asn  Leu His Glu 
    1190                 1195                 1200             


Asn Asn  Thr His Asn Gln Glu  Lys Lys Ile Gln Glu  Glu Ile Glu 
    1205                 1210                 1215             


Lys Lys  Glu Thr Leu Ile Gln  Glu Asn Val Val Leu  Pro Gln Ile 
    1220                 1225                 1230             


His Thr  Val Thr Gly Thr Lys  Asn Phe Met Lys Asn  Leu Phe Leu 
    1235                 1240                 1245             


Leu Ser  Thr Arg Gln Asn Val  Glu Gly Ser Tyr Asp  Gly Ala Tyr 
    1250                 1255                 1260             


Ala Pro  Val Leu Gln Asp Phe  Arg Ser Leu Asn Asp  Ser Thr Asn 
    1265                 1270                 1275             


Arg Thr  Lys Lys His Thr Ala  His Phe Ser Lys Lys  Gly Glu Glu 
    1280                 1285                 1290             


Glu Asn  Leu Glu Gly Leu Gly  Asn Gln Thr Lys Gln  Ile Val Glu 
    1295                 1300                 1305             


Lys Tyr  Ala Cys Thr Thr Arg  Ile Ser Pro Asn Thr  Ser Gln Gln 
    1310                 1315                 1320             


Asn Phe  Val Thr Gln Arg Ser  Lys Arg Ala Leu Lys  Gln Phe Arg 
    1325                 1330                 1335             


Leu Pro  Leu Glu Glu Thr Glu  Leu Glu Lys Arg Ile  Ile Val Asp 
    1340                 1345                 1350             


Asp Thr  Ser Thr Gln Trp Ser  Lys Asn Met Lys His  Leu Thr Pro 
    1355                 1360                 1365             


Ser Thr  Leu Thr Gln Ile Asp  Tyr Asn Glu Lys Glu  Lys Gly Ala 
    1370                 1375                 1380             


Ile Thr  Gln Ser Pro Leu Ser  Asp Cys Leu Thr Arg  Ser His Ser 
    1385                 1390                 1395             


Ile Pro  Gln Ala Asn Arg Ser  Pro Leu Pro Ile Ala  Lys Val Ser 
    1400                 1405                 1410             


Ser Phe  Pro Ser Ile Arg Pro  Ile Tyr Leu Thr Arg  Val Leu Phe 
    1415                 1420                 1425             


Gln Asp  Asn Ser Ser His Leu  Pro Ala Ala Ser Tyr  Arg Lys Lys 
    1430                 1435                 1440             


Asp Ser  Gly Val Gln Glu Ser  Ser His Phe Leu Gln  Gly Ala Lys 
    1445                 1450                 1455             


Lys Asn  Asn Leu Ser Leu Ala  Ile Leu Thr Leu Glu  Met Thr Gly 
    1460                 1465                 1470             


Asp Gln  Arg Glu Val Gly Ser  Leu Gly Thr Ser Ala  Thr Asn Ser 
    1475                 1480                 1485             


Val Thr  Tyr Lys Lys Val Glu  Asn Thr Val Leu Pro  Lys Pro Asp 
    1490                 1495                 1500             


Leu Pro  Lys Thr Ser Gly Lys  Val Glu Leu Leu Pro  Lys Val His 
    1505                 1510                 1515             


Ile Tyr  Gln Lys Asp Leu Phe  Pro Thr Glu Thr Ser  Asn Gly Ser 
    1520                 1525                 1530             


Pro Gly  His Leu Asp Leu Val  Glu Gly Ser Leu Leu  Gln Gly Thr 
    1535                 1540                 1545             


Glu Gly  Ala Ile Lys Trp Asn  Glu Ala Asn Arg Pro  Gly Lys Val 
    1550                 1555                 1560             


Pro Phe  Leu Arg Val Ala Thr  Glu Ser Ser Ala Lys  Thr Pro Ser 
    1565                 1570                 1575             


Lys Leu  Leu Asp Pro Leu Ala  Trp Asp Asn His Tyr  Gly Thr Gln 
    1580                 1585                 1590             


Ile Pro  Lys Glu Glu Trp Lys  Ser Gln Glu Lys Ser  Pro Glu Lys 
    1595                 1600                 1605             


Thr Ala  Phe Lys Lys Lys Asp  Thr Ile Leu Ser Leu  Asn Ala Cys 
    1610                 1615                 1620             


Glu Ser  Asn His Ala Ile Ala  Ala Ile Asn Glu Gly  Gln Asn Lys 
    1625                 1630                 1635             


Pro Glu  Ile Glu Val Thr Trp  Ala Lys Gln Gly Arg  Thr Glu Arg 
    1640                 1645                 1650             


Leu Cys  Ser Gln Asn Pro Pro  Val Leu Lys Arg His  Gln Arg Glu 
    1655                 1660                 1665             


Ile Thr  Arg Thr Thr Leu Gln  Ser Asp Gln Glu Glu  Ile Asp Tyr 
    1670                 1675                 1680             


Asp Asp  Thr Ile Ser Val Glu  Met Lys Lys Glu Asp  Phe Asp Ile 
    1685                 1690                 1695             


Tyr Asp  Glu Asp Glu Asn Gln  Ser Pro Arg Ser Phe  Gln Lys Lys 
    1700                 1705                 1710             


Thr Arg  His Tyr Phe Ile Ala  Ala Val Glu Arg Leu  Trp Asp Tyr 
    1715                 1720                 1725             


Gly Met  Ser Ser Ser Pro His  Val Leu Arg Asn Arg  Ala Gln Ser 
    1730                 1735                 1740             


Gly Ser  Val Pro Gln Phe Lys  Lys Val Val Phe Gln  Glu Phe Thr 
    1745                 1750                 1755             


Asp Gly  Ser Phe Thr Gln Pro  Leu Tyr Arg Gly Glu  Leu Asn Glu 
    1760                 1765                 1770             


His Leu  Gly Leu Leu Gly Pro  Tyr Ile Arg Ala Glu  Val Glu Asp 
    1775                 1780                 1785             


Asn Ile  Met Val Thr Phe Arg  Asn Gln Ala Ser Arg  Pro Tyr Ser 
    1790                 1795                 1800             


Phe Tyr  Ser Ser Leu Ile Ser  Tyr Glu Glu Asp Gln  Arg Gln Gly 
    1805                 1810                 1815             


Ala Glu  Pro Arg Lys Asn Phe  Val Lys Pro Asn Glu  Thr Lys Thr 
    1820                 1825                 1830             


Tyr Phe  Trp Lys Val Gln His  His Met Ala Pro Thr  Lys Asp Glu 
    1835                 1840                 1845             


Phe Asp  Cys Lys Ala Trp Ala  Tyr Phe Ser Asp Val  Asp Leu Glu 
    1850                 1855                 1860             


Lys Asp  Val His Ser Gly Leu  Ile Gly Pro Leu Leu  Val Cys His 
    1865                 1870                 1875             


Thr Asn  Thr Leu Asn Pro Ala  His Gly Arg Gln Val  Thr Val Gln 
    1880                 1885                 1890             


Glu Phe  Ala Leu Phe Phe Thr  Ile Phe Asp Glu Thr  Lys Ser Trp 
    1895                 1900                 1905             


Tyr Phe  Thr Glu Asn Met Glu  Arg Asn Cys Arg Ala  Pro Cys Asn 
    1910                 1915                 1920             


Ile Gln  Met Glu Asp Pro Thr  Phe Lys Glu Asn Tyr  Arg Phe His 
    1925                 1930                 1935             


Ala Ile  Asn Gly Tyr Ile Met  Asp Thr Leu Pro Gly  Leu Val Met 
    1940                 1945                 1950             


Ala Gln  Asp Gln Arg Ile Arg  Trp Tyr Leu Leu Ser  Met Gly Ser 
    1955                 1960                 1965             


Asn Glu  Asn Ile His Ser Ile  His Phe Ser Gly His  Val Phe Thr 
    1970                 1975                 1980             


Val Arg  Lys Lys Glu Glu Tyr  Lys Met Ala Leu Tyr  Asn Leu Tyr 
    1985                 1990                 1995             


Pro Gly  Val Phe Glu Thr Val  Glu Met Leu Pro Ser  Lys Ala Gly 
    2000                 2005                 2010             


Ile Trp  Arg Val Glu Cys Leu  Ile Gly Glu His Leu  His Ala Gly 
    2015                 2020                 2025             


Met Ser  Thr Leu Phe Leu Val  Tyr Ser Asn Lys Cys  Gln Thr Pro 
    2030                 2035                 2040             


Leu Gly  Met Ala Ser Gly His  Ile Arg Asp Phe Gln  Ile Thr Ala 
    2045                 2050                 2055             


Ser Gly  Gln Tyr Gly Gln Trp  Ala Pro Lys Leu Ala  Arg Leu His 
    2060                 2065                 2070             


Tyr Ser  Gly Ser Ile Asn Ala  Trp Ser Thr Lys Glu  Pro Phe Ser 
    2075                 2080                 2085             


Trp Ile  Lys Val Asp Leu Leu  Ala Pro Met Ile Ile  His Gly Ile 
    2090                 2095                 2100             


Lys Thr  Gln Gly Ala Arg Gln  Lys Phe Ser Ser Leu  Tyr Ile Ser 
    2105                 2110                 2115             


Gln Phe  Ile Ile Met Tyr Ser  Leu Asp Gly Lys Lys  Trp Gln Thr 
    2120                 2125                 2130             


Tyr Arg  Gly Asn Ser Thr Gly  Thr Leu Met Val Phe  Phe Gly Asn 
    2135                 2140                 2145             


Val Asp  Ser Ser Gly Ile Lys  His Asn Ile Phe Asn  Pro Pro Ile 
    2150                 2155                 2160             


Ile Ala  Arg Tyr Ile Arg Leu  His Pro Thr His Tyr  Ser Ile Arg 
    2165                 2170                 2175             


Ser Thr  Leu Arg Met Glu Leu  Met Gly Cys Asp Leu  Asn Ser Cys 
    2180                 2185                 2190             


Ser Met  Pro Leu Gly Met Glu  Ser Lys Ala Ile Ser  Asp Ala Gln 
    2195                 2200                 2205             


Ile Thr  Ala Ser Ser Tyr Phe  Thr Asn Met Phe Ala  Thr Trp Ser 
    2210                 2215                 2220             


Pro Ser  Lys Ala Arg Leu His  Leu Gln Gly Arg Ser  Asn Ala Trp 
    2225                 2230                 2235             


Arg Pro  Gln Val Asn Asn Pro  Lys Glu Trp Leu Gln  Val Asp Phe 
    2240                 2245                 2250             


Gln Lys  Thr Met Lys Val Thr  Gly Val Thr Thr Gln  Gly Val Lys 
    2255                 2260                 2265             


Ser Leu  Leu Thr Ser Met Tyr  Val Lys Glu Phe Leu  Ile Ser Ser 
    2270                 2275                 2280             


Ser Gln  Asp Gly His Gln Trp  Thr Leu Phe Phe Gln  Asn Gly Lys 
    2285                 2290                 2295             


Val Lys  Val Phe Gln Gly Asn  Gln Asp Ser Phe Thr  Pro Val Val 
    2300                 2305                 2310             


Asn Ser  Leu Asp Pro Pro Leu  Leu Thr Arg Tyr Leu  Arg Ile His 
    2315                 2320                 2325             


Pro Gln  Ser Trp Val His Gln  Ile Ala Leu Arg Met  Glu Val Leu 
    2330                 2335                 2340             


Gly Cys  Glu Ala Gln Asp Leu  Tyr 
    2345                 2350     


<210>  2
<211>  216
<212>  PRT
<213>  Homo sapiens

<400>  2

Met Arg Ile Gln Asp Pro Gly Lys Val Phe Phe Gly Asn Val Asp Ser 
1               5                   10                  15      


Ser Gly Ile Lys His Asn Ile Phe Asn Pro Pro Ile Ile Ala Arg Tyr 
            20                  25                  30          


Ile Arg Leu His Pro Thr His Tyr Ser Ile Arg Ser Thr Leu Arg Met 
        35                  40                  45              


Glu Leu Met Gly Cys Asp Leu Asn Ser Cys Ser Met Pro Leu Gly Met 
    50                  55                  60                  


Glu Ser Lys Ala Ile Ser Asp Ala Gln Ile Thr Ala Ser Ser Tyr Phe 
65                  70                  75                  80  


Thr Asn Met Phe Ala Thr Trp Ser Pro Ser Lys Ala Arg Leu His Leu 
                85                  90                  95      


Gln Gly Arg Ser Asn Ala Trp Arg Pro Gln Val Asn Asn Pro Lys Glu 
            100                 105                 110         


Trp Leu Gln Val Asp Phe Gln Lys Thr Met Lys Val Thr Gly Val Thr 
        115                 120                 125             


Thr Gln Gly Val Lys Ser Leu Leu Thr Ser Met Tyr Val Lys Glu Phe 
    130                 135                 140                 


Leu Ile Ser Ser Ser Gln Asp Gly His Gln Trp Thr Leu Phe Phe Gln 
145                 150                 155                 160 


Asn Gly Lys Val Lys Val Phe Gln Gly Asn Gln Asp Ser Phe Thr Pro 
                165                 170                 175     


Val Val Asn Ser Leu Asp Pro Pro Leu Leu Thr Arg Tyr Leu Arg Ile 
            180                 185                 190         


His Pro Gln Ser Trp Val His Gln Ile Ala Leu Arg Met Glu Val Leu 
        195                 200                 205             


Gly Cys Glu Ala Gln Asp Leu Tyr 
    210                 215     


<210>  3
<211>  5213
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Hybrid FVIII

<400>  3
gtgtcgtgaa aactacccct aaaagccacc ggcgtggcta gcatgcaaat agagctctcc       60

acctgcttct ttctgtgcct tttgcgattc tgctttagtg ccatcaggag atactacctg      120

ggcgcagtgg aactgtcctg ggactaccgg caaagtgaac tcctccgtga gctgcacgtg      180

gacaccagat ttcctgctac agcgccagga gctcttccgt tgggcccgtc agtcctgtac      240

aaaaagactg tgttcgtaga gttcacggat caacttttca gcgttgccag gcccaggcca      300

ccatggatgg gtctgctggg tcctaccatc caggctgagg tttacgacac ggtggtcgtt      360

accctgaaga acatggcttc tcatcccgtt agtcttcacg ctgtcggcgt ctccttctgg      420

aaatcttccg aaggcgctga atatgaggat cacaccagcc aaagggagaa ggaagacgat      480

aaagtccttc ccggtaaaag ccaaacctac gtctggcagg tcctgaaaga aaatggtcca      540

acagcctctg acccaccatg tctcacctac tcatacctgt ctcacgtgga cctggtgaaa      600

gacctgaatt cgggcctcat tggagccctg ctggtttgta gagaagggag tctgaccaga      660

gaaaggaccc agaacctgca cgaatttgta ctactttttg ctgtctttga tgaagggaaa      720

agttggcact cagcaagaaa tgactcctgg acacgggcca tggatcccgc acctgccagg      780

gcccagcctg caatgcacac agtcaatggc tatgtcaaca ggtctctgcc aggtctgatc      840

ggatgtcata agaaatcagt ctactggcac gtgattggaa tgggcaccag cccggaagtg      900

cactccattt ttcttgaagg ccacacgttt ctcgtgaggc accatcgcca ggcttccttg      960

gagatctcgc cactaacttt cctcactgct cagacattcc tgatggacct tggccagttc     1020

ctactgtttt gtcatatctc ttcccaccac catggtggca tggaggctca cgtcagagta     1080

gaaagctgcg ccgaggagcc ccagctgcgg aggaaagctg atgaagagga agattatgat     1140

gacaatttgt acgactcgga catggacgtg gtccggctcg atggtgacga cgtgtctccc     1200

tttatccaaa tccgctcagt tgccaagaag catcctaaaa cttgggtaca ttacattgct     1260

gctgaagagg aggactggga ctatgctccc ttagtcctcg cccccgatga cagaagttat     1320

aaaagtcaat atttgaacaa tggccctcag cggattggta ggaagtacaa aaaagtccga     1380

tttatggcat acacagatga aacctttaag actcgtgaag ctattcagca tgaatcagga     1440

atcttgggac ctttacttta tggggaagtt ggagacacac tgttgattat atttaagaat     1500

caagcaagca gaccatataa catctaccct cacggaatca ctgatgtccg tcctttgtat     1560

tcaaggagat taccaaaagg tgtaaaacat ttgaaggatt ttccaattct gccaggagaa     1620

atattcaaat ataaatggac agtgactgta gaagatgggc caactaaatc agatcctcgg     1680

tgcctgaccc gctattactc tagtttcgtt aatatggaga gagatctagc ttcaggactc     1740

attggccctc tcctcatctg ctacaaagaa tctgtagatc aaagaggaaa ccagataatg     1800

tcagacaaga ggaatgtcat cctgttttct gtatttgatg agaaccgaag ctggtacctc     1860

acagagaata tacaacgctt tctccccaat ccagctggag tgcagcttga ggatccagag     1920

ttccaagcct ccaacatcat gcacagcatc aatggctatg tttttgatag tttgcagttg     1980

tcagtttgtt tgcatgaggt ggcatactgg tacattctaa gcattggagc acagactgac     2040

ttcctttctg tcttcttctc tggatatacc ttcaaacaca aaatggtcta tgaagacaca     2100

ctcaccctat tcccattctc aggagaaact gtcttcatgt cgatggaaaa cccaggtcta     2160

tggattctgg ggtgccacaa ctcagacttt cggaacagag gcatgaccgc cttactgaag     2220

gtttctagtt gtgacaagaa cactggtgat tattacgagg acagttatga agatatttca     2280

gcatacttgc tgagtaaaaa caatgccatt gaaccaagaa gcttctccca gaattcaaga     2340

caccctagca ctaggcaaaa gcaatttaat gccaccacaa ttccagaaaa tgacatagag     2400

aagactgacc cttggtttgc acacagaaca cctatgccta aaatacaaaa tgtctcctct     2460

agtgatttgt tgatgctctt gcgacagagt cctactccac atgggctatc cttatctgat     2520

ctccaagaag ccaaatatga gactttttct gatgatccat cacctggagc aatagacagt     2580

aataacagcc tgtctgaaat gacacacttc aggccacagc tccatcacag tggggacatg     2640

gtatttaccc ctgagtcagg cctccaatta agattaaatg agaaactggg gacaactgca     2700

gcaacagagt tgaagaaact tgatttcaaa gtttctagta catcaaataa tctgatttca     2760

acaattccat cagacaattt ggcagcaggt actgataata caagttcctt aggaccccca     2820

agtatgccag ttcattatga tagtcaatta gataccactc tatttggcaa aaagtcatct     2880

ccccttactg agtctggtgg acctctgagc ttgagtgaag aaaataatga ttcaaagttg     2940

ttagaatcag gtttaatgaa tagccaagaa agttcatggg gaaaaaatgt atcgtcaaca     3000

gagagtggta ggttatttaa agggaaaaga gctcatggac ctgctttgtt gactaaagat     3060

gggaggactg aaaggctgtg ctctcaaaac ccaccagtct tgaaacgcca tcaacgggaa     3120

ataactcgta ctactcttca gtcagatcaa gaggaaattg actatgatga taccatatca     3180

gttgaaatga agaaggaaga ttttgacatt tatgatgagg atgaaaatca gagcccccgc     3240

agctttcaga agagaacccg acactatttc attgctgcgg tggagcagct ctgggattac     3300

gggatgagcg aatccccccg ggcgctaaga aacagggctc agaacggaga ggtgcctcgg     3360

ttcaagaagg tggtcttccg ggaatttgct gacggctcct tcacgcagcc gtcgtaccgc     3420

ggggaactca acaaacactt ggggctcttg ggaccctaca tcagagcgga agttgaagac     3480

aacatcatgg taactttcaa aaaccaggcg tctcgtccct attccttcta ctcgagcctt     3540

atttcttatc cggatgatca ggagcaaggg gcagaacctc gacacaactt cgtccagcca     3600

aatgaaacca gaacttactt ttggaaagtg cagcatcaca tggcacccac agaagacgag     3660

tttgactgca aagcctgggc ctacttttct gatgttgacc tggaaaaaga tgtgcactca     3720

ggcttgatcg gcccccttct gatctgccgc gccaacaccc tgaacgctgc tcacggtaga     3780

caagtgaccg tgcaagaatt tgctctgttt ttcactattt ttgatgagac aaagagctgg     3840

tacttcactg aaaatgtgga aaggaactgc cgggccccct gccacctgca gatggaggac     3900

cccactctga aagaaaacta tcgcttccat gcaatcaatg gctatgtgat ggatacactc     3960

cctggcttag taatggctca gaatcaaagg atccgatggt atctgctcag catgggcagc     4020

aatgaaaata tccattcgat tcattttagc ggacacgtgt tcagtgtacg gaaaaaggag     4080

gagtataaaa tggccgtgta caatctctat ccgggtgtct ttgagacagt ggaaatgcta     4140

ccgtccaaag ttggaatttg gcgaatagaa tgcctgattg gcgagcacct gcaagctggg     4200

atgagcacga ctttcctggt gtacagcaag aagtgtcaga ctcccctggg aatggcttct     4260

ggacacatta gagattttca gattacagct tcaggacaat atggacagtg ggccccaaag     4320

ctggccagac ttcattattc cggatcaatc aatgcctgga gcaccaagga gcccttttct     4380

tggatcaagg tggatctgtt ggcaccaatg attattcacg gcatcaagac ccagggtgcc     4440

cgtcagaagt tctccagcct ctacatctct cagtttatca tcatgtatag tcttgatggg     4500

aagaagtggc agacttatcg aggaaattcc actggaacct taatggtctt ctttggcaat     4560

gtggattcat ctgggataaa acacaatatt tttaaccctc caattattgc tcgatacatc     4620

cgtttgcacc caactcatta tagcattcgc agcactcttc gcatggagtt gatgggctgt     4680

gatttaaata gttgcagcat gccattggga atggagagta aagcaatatc agatgcacag     4740

attactgctt catcctactt taccaatatg tttgccacct ggtctccttc aaaagctcga     4800

cttcacctcc aagggaggag taatgcctgg agacctcagg tgaataatcc aaaagagtgg     4860

ctgcaagtgg acttccagaa gacaatgaaa gtcacaggag taactactca gggagtaaaa     4920

tctctgctta ccagcatgta tgtgaaggag ttcctcatct ccagcagtca agatggccat     4980

cagtggactc tcttttttca gaatggcaaa gtaaaggttt ttcagggaaa tcaagactcc     5040

ttcacacctg tggtgaactc tctagaccca ccgttactga ctcgctacct tcgaattcac     5100

ccccagagtt gggtgcacca gattgccctg aggatggagg ttctgggctg cgaggcacag     5160

gacctctact gagctcgagt ctagagggcc cgtttaaacc cgctgatcag cct            5213


<210>  4
<211>  1709
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Hybrid FVIII

<400>  4

Met Gln Ile Glu Leu Ser Thr Cys Phe Phe Leu Cys Leu Leu Arg Phe 
1               5                   10                  15      


Cys Phe Ser Ala Ile Arg Arg Tyr Tyr Leu Gly Ala Val Glu Leu Ser 
            20                  25                  30          


Trp Asp Tyr Arg Gln Ser Glu Leu Leu Arg Glu Leu His Val Asp Thr 
        35                  40                  45              


Arg Phe Pro Ala Thr Ala Pro Gly Ala Leu Pro Leu Gly Pro Ser Val 
    50                  55                  60                  


Leu Tyr Lys Lys Thr Val Phe Val Glu Phe Thr Asp Gln Leu Phe Ser 
65                  70                  75                  80  


Val Ala Arg Pro Arg Pro Pro Trp Met Gly Leu Leu Gly Pro Thr Ile 
                85                  90                  95      


Gln Ala Glu Val Tyr Asp Thr Val Val Val Thr Leu Lys Asn Met Ala 
            100                 105                 110         


Ser His Pro Val Ser Leu His Ala Val Gly Val Ser Phe Trp Lys Ser 
        115                 120                 125             


Ser Glu Gly Ala Glu Tyr Glu Asp His Thr Ser Gln Arg Glu Lys Glu 
    130                 135                 140                 


Asp Asp Lys Val Leu Pro Gly Lys Ser Gln Thr Tyr Val Trp Gln Val 
145                 150                 155                 160 


Leu Lys Glu Asn Gly Pro Thr Ala Ser Asp Pro Pro Cys Leu Thr Tyr 
                165                 170                 175     


Ser Tyr Leu Ser His Val Asp Leu Val Lys Asp Leu Asn Ser Gly Leu 
            180                 185                 190         


Ile Gly Ala Leu Leu Val Cys Arg Glu Gly Ser Leu Thr Arg Glu Arg 
        195                 200                 205             


Thr Gln Asn Leu His Glu Phe Val Leu Leu Phe Ala Val Phe Asp Glu 
    210                 215                 220                 


Gly Lys Ser Trp His Ser Ala Arg Asn Asp Ser Trp Thr Arg Ala Met 
225                 230                 235                 240 


Asp Pro Ala Pro Ala Arg Ala Gln Pro Ala Met His Thr Val Asn Gly 
                245                 250                 255     


Tyr Val Asn Arg Ser Leu Pro Gly Leu Ile Gly Cys His Lys Lys Ser 
            260                 265                 270         


Val Tyr Trp His Val Ile Gly Met Gly Thr Ser Pro Glu Val His Ser 
        275                 280                 285             


Ile Phe Leu Glu Gly His Thr Phe Leu Val Arg His His Arg Gln Ala 
    290                 295                 300                 


Ser Leu Glu Ile Ser Pro Leu Thr Phe Leu Thr Ala Gln Thr Phe Leu 
305                 310                 315                 320 


Met Asp Leu Gly Gln Phe Leu Leu Phe Cys His Ile Ser Ser His His 
                325                 330                 335     


His Gly Gly Met Glu Ala His Val Arg Val Glu Ser Cys Ala Glu Glu 
            340                 345                 350         


Pro Gln Leu Arg Arg Lys Ala Asp Glu Glu Glu Asp Tyr Asp Asp Asn 
        355                 360                 365             


Leu Tyr Asp Ser Asp Met Asp Val Val Arg Leu Asp Gly Asp Asp Val 
    370                 375                 380                 


Ser Pro Phe Ile Gln Ile Arg Ser Val Ala Lys Lys His Pro Lys Thr 
385                 390                 395                 400 


Trp Val His Tyr Ile Ala Ala Glu Glu Glu Asp Trp Asp Tyr Ala Pro 
                405                 410                 415     


Leu Val Leu Ala Pro Asp Asp Arg Ser Tyr Lys Ser Gln Tyr Leu Asn 
            420                 425                 430         


Asn Gly Pro Gln Arg Ile Gly Arg Lys Tyr Lys Lys Val Arg Phe Met 
        435                 440                 445             


Ala Tyr Thr Asp Glu Thr Phe Lys Thr Arg Glu Ala Ile Gln His Glu 
    450                 455                 460                 


Ser Gly Ile Leu Gly Pro Leu Leu Tyr Gly Glu Val Gly Asp Thr Leu 
465                 470                 475                 480 


Leu Ile Ile Phe Lys Asn Gln Ala Ser Arg Pro Tyr Asn Ile Tyr Pro 
                485                 490                 495     


His Gly Ile Thr Asp Val Arg Pro Leu Tyr Ser Arg Arg Leu Pro Lys 
            500                 505                 510         


Gly Val Lys His Leu Lys Asp Phe Pro Ile Leu Pro Gly Glu Ile Phe 
        515                 520                 525             


Lys Tyr Lys Trp Thr Val Thr Val Glu Asp Gly Pro Thr Lys Ser Asp 
    530                 535                 540                 


Pro Arg Cys Leu Thr Arg Tyr Tyr Ser Ser Phe Val Asn Met Glu Arg 
545                 550                 555                 560 


Asp Leu Ala Ser Gly Leu Ile Gly Pro Leu Leu Ile Cys Tyr Lys Glu 
                565                 570                 575     


Ser Val Asp Gln Arg Gly Asn Gln Ile Met Ser Asp Lys Arg Asn Val 
            580                 585                 590         


Ile Leu Phe Ser Val Phe Asp Glu Asn Arg Ser Trp Tyr Leu Thr Glu 
        595                 600                 605             


Asn Ile Gln Arg Phe Leu Pro Asn Pro Ala Gly Val Gln Leu Glu Asp 
    610                 615                 620                 


Pro Glu Phe Gln Ala Ser Asn Ile Met His Ser Ile Asn Gly Tyr Val 
625                 630                 635                 640 


Phe Asp Ser Leu Gln Leu Ser Val Cys Leu His Glu Val Ala Tyr Trp 
                645                 650                 655     


Tyr Ile Leu Ser Ile Gly Ala Gln Thr Asp Phe Leu Ser Val Phe Phe 
            660                 665                 670         


Ser Gly Tyr Thr Phe Lys His Lys Met Val Tyr Glu Asp Thr Leu Thr 
        675                 680                 685             


Leu Phe Pro Phe Ser Gly Glu Thr Val Phe Met Ser Met Glu Asn Pro 
    690                 695                 700                 


Gly Leu Trp Ile Leu Gly Cys His Asn Ser Asp Phe Arg Asn Arg Gly 
705                 710                 715                 720 


Met Thr Ala Leu Leu Lys Val Ser Ser Cys Asp Lys Asn Thr Gly Asp 
                725                 730                 735     


Tyr Tyr Glu Asp Ser Tyr Glu Asp Ile Ser Ala Tyr Leu Leu Ser Lys 
            740                 745                 750         


Asn Asn Ala Ile Glu Pro Arg Ser Phe Ser Gln Asn Ser Arg His Pro 
        755                 760                 765             


Ser Thr Arg Gln Lys Gln Phe Asn Ala Thr Thr Ile Pro Glu Asn Asp 
    770                 775                 780                 


Ile Glu Lys Thr Asp Pro Trp Phe Ala His Arg Thr Pro Met Pro Lys 
785                 790                 795                 800 


Ile Gln Asn Val Ser Ser Ser Asp Leu Leu Met Leu Leu Arg Gln Ser 
                805                 810                 815     


Pro Thr Pro His Gly Leu Ser Leu Ser Asp Leu Gln Glu Ala Lys Tyr 
            820                 825                 830         


Glu Thr Phe Ser Asp Asp Pro Ser Pro Gly Ala Ile Asp Ser Asn Asn 
        835                 840                 845             


Ser Leu Ser Glu Met Thr His Phe Arg Pro Gln Leu His His Ser Gly 
    850                 855                 860                 


Asp Met Val Phe Thr Pro Glu Ser Gly Leu Gln Leu Arg Leu Asn Glu 
865                 870                 875                 880 


Lys Leu Gly Thr Thr Ala Ala Thr Glu Leu Lys Lys Leu Asp Phe Lys 
                885                 890                 895     


Val Ser Ser Thr Ser Asn Asn Leu Ile Ser Thr Ile Pro Ser Asp Asn 
            900                 905                 910         


Leu Ala Ala Gly Thr Asp Asn Thr Ser Ser Leu Gly Pro Pro Ser Met 
        915                 920                 925             


Pro Val His Tyr Asp Ser Gln Leu Asp Thr Thr Leu Phe Gly Lys Lys 
    930                 935                 940                 


Ser Ser Pro Leu Thr Glu Ser Gly Gly Pro Leu Ser Leu Ser Glu Glu 
945                 950                 955                 960 


Asn Asn Asp Ser Lys Leu Leu Glu Ser Gly Leu Met Asn Ser Gln Glu 
                965                 970                 975     


Ser Ser Trp Gly Lys Asn Val Ser Ser Thr Glu Ser Gly Arg Leu Phe 
            980                 985                 990         


Lys Gly Lys Arg Ala His Gly Pro  Ala Leu Leu Thr Lys  Asp Gly Arg 
        995                 1000                 1005             


Thr Glu  Arg Leu Cys Ser Gln  Asn Pro Pro Val Leu  Lys Arg His 
    1010                 1015                 1020             


Gln Arg  Glu Ile Thr Arg Thr  Thr Leu Gln Ser Asp  Gln Glu Glu 
    1025                 1030                 1035             


Ile Asp  Tyr Asp Asp Thr Ile  Ser Val Glu Met Lys  Lys Glu Asp 
    1040                 1045                 1050             


Phe Asp  Ile Tyr Asp Glu Asp  Glu Asn Gln Ser Pro  Arg Ser Phe 
    1055                 1060                 1065             


Gln Lys  Arg Thr Arg His Tyr  Phe Ile Ala Ala Val  Glu Gln Leu 
    1070                 1075                 1080             


Trp Asp  Tyr Gly Met Ser Glu  Ser Pro Arg Ala Leu  Arg Asn Arg 
    1085                 1090                 1095             


Ala Gln  Asn Gly Glu Val Pro  Arg Phe Lys Lys Val  Val Phe Arg 
    1100                 1105                 1110             


Glu Phe  Ala Asp Gly Ser Phe  Thr Gln Pro Ser Tyr  Arg Gly Glu 
    1115                 1120                 1125             


Leu Asn  Lys His Leu Gly Leu  Leu Gly Pro Tyr Ile  Arg Ala Glu 
    1130                 1135                 1140             


Val Glu  Asp Asn Ile Met Val  Thr Phe Lys Asn Gln  Ala Ser Arg 
    1145                 1150                 1155             


Pro Tyr  Ser Phe Tyr Ser Ser  Leu Ile Ser Tyr Pro  Asp Asp Gln 
    1160                 1165                 1170             


Glu Gln  Gly Ala Glu Pro Arg  His Asn Phe Val Gln  Pro Asn Glu 
    1175                 1180                 1185             


Thr Arg  Thr Tyr Phe Trp Lys  Val Gln His His Met  Ala Pro Thr 
    1190                 1195                 1200             


Glu Asp  Glu Phe Asp Cys Lys  Ala Trp Ala Tyr Phe  Ser Asp Val 
    1205                 1210                 1215             


Asp Leu  Glu Lys Asp Val His  Ser Gly Leu Ile Gly  Pro Leu Leu 
    1220                 1225                 1230             


Ile Cys  Arg Ala Asn Thr Leu  Asn Ala Ala His Gly  Arg Gln Val 
    1235                 1240                 1245             


Thr Val  Gln Glu Phe Ala Leu  Phe Phe Thr Ile Phe  Asp Glu Thr 
    1250                 1255                 1260             


Lys Ser  Trp Tyr Phe Thr Glu  Asn Val Glu Arg Asn  Cys Arg Ala 
    1265                 1270                 1275             


Pro Cys  His Leu Gln Met Glu  Asp Pro Thr Leu Lys  Glu Asn Tyr 
    1280                 1285                 1290             


Arg Phe  His Ala Ile Asn Gly  Tyr Val Met Asp Thr  Leu Pro Gly 
    1295                 1300                 1305             


Leu Val  Met Ala Gln Asn Gln  Arg Ile Arg Trp Tyr  Leu Leu Ser 
    1310                 1315                 1320             


Met Gly  Ser Asn Glu Asn Ile  His Ser Ile His Phe  Ser Gly His 
    1325                 1330                 1335             


Val Phe  Ser Val Arg Lys Lys  Glu Glu Tyr Lys Met  Ala Val Tyr 
    1340                 1345                 1350             


Asn Leu  Tyr Pro Gly Val Phe  Glu Thr Val Glu Met  Leu Pro Ser 
    1355                 1360                 1365             


Lys Val  Gly Ile Trp Arg Ile  Glu Cys Leu Ile Gly  Glu His Leu 
    1370                 1375                 1380             


Gln Ala  Gly Met Ser Thr Thr  Phe Leu Val Tyr Ser  Lys Lys Cys 
    1385                 1390                 1395             


Gln Thr  Pro Leu Gly Met Ala  Ser Gly His Ile Arg  Asp Phe Gln 
    1400                 1405                 1410             


Ile Thr  Ala Ser Gly Gln Tyr  Gly Gln Trp Ala Pro  Lys Leu Ala 
    1415                 1420                 1425             


Arg Leu  His Tyr Ser Gly Ser  Ile Asn Ala Trp Ser  Thr Lys Glu 
    1430                 1435                 1440             


Pro Phe  Ser Trp Ile Lys Val  Asp Leu Leu Ala Pro  Met Ile Ile 
    1445                 1450                 1455             


His Gly  Ile Lys Thr Gln Gly  Ala Arg Gln Lys Phe  Ser Ser Leu 
    1460                 1465                 1470             


Tyr Ile  Ser Gln Phe Ile Ile  Met Tyr Ser Leu Asp  Gly Lys Lys 
    1475                 1480                 1485             


Trp Gln  Thr Tyr Arg Gly Asn  Ser Thr Gly Thr Leu  Met Val Phe 
    1490                 1495                 1500             


Phe Gly  Asn Val Asp Ser Ser  Gly Ile Lys His Asn  Ile Phe Asn 
    1505                 1510                 1515             


Pro Pro  Ile Ile Ala Arg Tyr  Ile Arg Leu His Pro  Thr His Tyr 
    1520                 1525                 1530             


Ser Ile  Arg Ser Thr Leu Arg  Met Glu Leu Met Gly  Cys Asp Leu 
    1535                 1540                 1545             


Asn Ser  Cys Ser Met Pro Leu  Gly Met Glu Ser Lys  Ala Ile Ser 
    1550                 1555                 1560             


Asp Ala  Gln Ile Thr Ala Ser  Ser Tyr Phe Thr Asn  Met Phe Ala 
    1565                 1570                 1575             


Thr Trp  Ser Pro Ser Lys Ala  Arg Leu His Leu Gln  Gly Arg Ser 
    1580                 1585                 1590             


Asn Ala  Trp Arg Pro Gln Val  Asn Asn Pro Lys Glu  Trp Leu Gln 
    1595                 1600                 1605             


Val Asp  Phe Gln Lys Thr Met  Lys Val Thr Gly Val  Thr Thr Gln 
    1610                 1615                 1620             


Gly Val  Lys Ser Leu Leu Thr  Ser Met Tyr Val Lys  Glu Phe Leu 
    1625                 1630                 1635             


Ile Ser  Ser Ser Gln Asp Gly  His Gln Trp Thr Leu  Phe Phe Gln 
    1640                 1645                 1650             


Asn Gly  Lys Val Lys Val Phe  Gln Gly Asn Gln Asp  Ser Phe Thr 
    1655                 1660                 1665             


Pro Val  Val Asn Ser Leu Asp  Pro Pro Leu Leu Thr  Arg Tyr Leu 
    1670                 1675                 1680             


Arg Ile  His Pro Gln Ser Trp  Val His Gln Ile Ala  Leu Arg Met 
    1685                 1690                 1695             


Glu Val  Leu Gly Cys Glu Ala  Gln Asp Leu Tyr 
    1700                 1705                 


<210>  5
<211>  1467
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Hybrid FVIII

<400>  5

Met Gln Leu Glu Leu Ser Thr Cys Val Phe Leu Cys Leu Leu Pro Leu 
1               5                   10                  15      


Gly Phe Ser Ala Ile Arg Arg Tyr Tyr Leu Gly Ala Val Glu Leu Ser 
            20                  25                  30          


Trp Asp Tyr Arg Gln Ser Glu Leu Leu Arg Glu Leu His Val Asp Thr 
        35                  40                  45              


Arg Phe Pro Ala Thr Ala Pro Gly Ala Leu Pro Leu Gly Pro Ser Val 
    50                  55                  60                  


Leu Tyr Lys Lys Thr Val Phe Val Glu Phe Thr Asp Gln Leu Phe Ser 
65                  70                  75                  80  


Val Ala Arg Pro Arg Pro Pro Trp Met Gly Leu Leu Gly Pro Thr Ile 
                85                  90                  95      


Gln Ala Glu Val Tyr Asp Thr Val Val Val Thr Leu Lys Asn Met Ala 
            100                 105                 110         


Ser His Pro Val Ser Leu His Ala Val Gly Val Ser Phe Trp Lys Ser 
        115                 120                 125             


Ser Glu Gly Ala Glu Tyr Glu Asp His Thr Ser Gln Arg Glu Lys Glu 
    130                 135                 140                 


Asp Asp Lys Val Leu Pro Gly Lys Ser Gln Thr Tyr Val Trp Gln Val 
145                 150                 155                 160 


Leu Lys Glu Asn Gly Pro Thr Ala Ser Asp Pro Pro Cys Leu Thr Tyr 
                165                 170                 175     


Ser Tyr Leu Ser His Val Asp Leu Val Lys Asp Leu Asn Ser Gly Leu 
            180                 185                 190         


Ile Gly Ala Leu Leu Val Cys Arg Glu Gly Ser Leu Thr Arg Glu Arg 
        195                 200                 205             


Thr Gln Asn Leu His Glu Phe Val Leu Leu Phe Ala Val Phe Asp Glu 
    210                 215                 220                 


Gly Lys Ser Trp His Ser Ala Arg Asn Asp Ser Trp Thr Arg Ala Met 
225                 230                 235                 240 


Asp Pro Ala Pro Ala Arg Ala Gln Pro Ala Met His Thr Val Asn Gly 
                245                 250                 255     


Tyr Val Asn Arg Ser Leu Pro Gly Leu Ile Gly Cys His Lys Lys Ser 
            260                 265                 270         


Val Tyr Trp His Val Ile Gly Met Gly Thr Ser Pro Glu Val His Ser 
        275                 280                 285             


Ile Phe Leu Glu Gly His Thr Phe Leu Val Arg His His Arg Gln Ala 
    290                 295                 300                 


Ser Leu Glu Ile Ser Pro Leu Thr Phe Leu Thr Ala Gln Thr Phe Leu 
305                 310                 315                 320 


Met Asp Leu Gly Gln Phe Leu Leu Phe Cys His Ile Ser Ser His His 
                325                 330                 335     


His Gly Gly Met Glu Ala His Val Arg Val Glu Ser Cys Ala Glu Glu 
            340                 345                 350         


Pro Gln Leu Arg Arg Lys Ala Asp Glu Glu Glu Asp Tyr Asp Asp Asn 
        355                 360                 365             


Leu Tyr Asp Ser Asp Met Asp Val Val Arg Leu Asp Gly Asp Asp Val 
    370                 375                 380                 


Ser Pro Phe Ile Gln Ile Arg Ser Val Ala Lys Lys His Pro Lys Thr 
385                 390                 395                 400 


Trp Val His Tyr Ile Ala Ala Glu Glu Glu Asp Trp Asp Tyr Ala Pro 
                405                 410                 415     


Leu Val Leu Ala Pro Asp Asp Arg Ser Tyr Lys Ser Gln Tyr Leu Asn 
            420                 425                 430         


Asn Gly Pro Gln Arg Ile Gly Arg Lys Tyr Lys Lys Val Arg Phe Met 
        435                 440                 445             


Ala Tyr Thr Asp Glu Thr Phe Lys Thr Arg Glu Ala Ile Gln His Glu 
    450                 455                 460                 


Ser Gly Ile Leu Gly Pro Leu Leu Tyr Gly Glu Val Gly Asp Thr Leu 
465                 470                 475                 480 


Leu Ile Ile Phe Lys Asn Gln Ala Ser Arg Pro Tyr Asn Ile Tyr Pro 
                485                 490                 495     


His Gly Ile Thr Asp Val Arg Pro Leu Tyr Ser Arg Arg Leu Pro Lys 
            500                 505                 510         


Gly Val Lys His Leu Lys Asp Phe Pro Ile Leu Pro Gly Glu Ile Phe 
        515                 520                 525             


Lys Tyr Lys Trp Thr Val Thr Val Glu Asp Gly Pro Thr Lys Ser Asp 
    530                 535                 540                 


Pro Arg Cys Leu Thr Arg Tyr Tyr Ser Ser Phe Val Asn Met Glu Arg 
545                 550                 555                 560 


Asp Leu Ala Ser Gly Leu Ile Gly Pro Leu Leu Ile Cys Tyr Lys Glu 
                565                 570                 575     


Ser Val Asp Gln Arg Gly Asn Gln Ile Met Ser Asp Lys Arg Asn Val 
            580                 585                 590         


Ile Leu Phe Ser Val Phe Asp Glu Asn Arg Ser Trp Tyr Leu Thr Glu 
        595                 600                 605             


Asn Ile Gln Arg Phe Leu Pro Asn Pro Ala Gly Val Gln Leu Glu Asp 
    610                 615                 620                 


Pro Glu Phe Gln Ala Ser Asn Ile Met His Ser Ile Asn Gly Tyr Val 
625                 630                 635                 640 


Phe Asp Ser Leu Gln Leu Ser Val Cys Leu His Glu Val Ala Tyr Trp 
                645                 650                 655     


Tyr Ile Leu Ser Ile Gly Ala Gln Thr Asp Phe Leu Ser Val Phe Phe 
            660                 665                 670         


Ser Gly Tyr Thr Phe Lys His Lys Met Val Tyr Glu Asp Thr Leu Thr 
        675                 680                 685             


Leu Phe Pro Phe Ser Gly Glu Thr Val Phe Met Ser Met Glu Asn Pro 
    690                 695                 700                 


Gly Leu Trp Ile Leu Gly Cys His Asn Ser Asp Phe Arg Asn Arg Gly 
705                 710                 715                 720 


Met Thr Ala Leu Leu Lys Val Ser Ser Cys Asp Lys Asn Thr Gly Asp 
                725                 730                 735     


Tyr Tyr Glu Asp Ser Tyr Glu Asp Ile Ser Ala Tyr Leu Leu Ser Lys 
            740                 745                 750         


Asn Asn Ala Ile Glu Pro Arg Ser Phe Ala Gln Asn Ser Arg Pro Pro 
        755                 760                 765             


Ser Ala Ser Ala Pro Lys Pro Pro Val Leu Arg Arg His Gln Arg Asp 
    770                 775                 780                 


Ile Ser Leu Pro Thr Phe Gln Pro Glu Glu Asp Lys Met Asp Tyr Asp 
785                 790                 795                 800 


Asp Ile Phe Ser Thr Glu Thr Lys Gly Glu Asp Phe Asp Ile Tyr Gly 
                805                 810                 815     


Glu Asp Glu Asn Gln Asp Pro Arg Ser Phe Gln Lys Arg Thr Arg His 
            820                 825                 830         


Tyr Phe Ile Ala Ala Val Glu Gln Leu Trp Asp Tyr Gly Met Ser Glu 
        835                 840                 845             


Ser Pro Arg Ala Leu Arg Asn Arg Ala Gln Asn Gly Glu Val Pro Arg 
    850                 855                 860                 


Phe Lys Lys Val Val Phe Arg Glu Phe Ala Asp Gly Ser Phe Thr Gln 
865                 870                 875                 880 


Pro Ser Tyr Arg Gly Glu Leu Asn Lys His Leu Gly Leu Leu Gly Pro 
                885                 890                 895     


Tyr Ile Arg Ala Glu Val Glu Asp Asn Ile Met Val Thr Phe Lys Asn 
            900                 905                 910         


Gln Ala Ser Arg Pro Tyr Ser Phe Tyr Ser Ser Leu Ile Ser Tyr Pro 
        915                 920                 925             


Asp Asp Gln Glu Gln Gly Ala Glu Pro Arg His Asn Phe Val Gln Pro 
    930                 935                 940                 


Asn Glu Thr Arg Thr Tyr Phe Trp Lys Val Gln His His Met Ala Pro 
945                 950                 955                 960 


Thr Glu Asp Glu Phe Asp Cys Lys Ala Trp Ala Tyr Phe Ser Asp Val 
                965                 970                 975     


Asp Leu Glu Lys Asp Val His Ser Gly Leu Ile Gly Pro Leu Leu Ile 
            980                 985                 990         


Cys Arg Ala Asn Thr Leu Asn Ala  Ala His Gly Arg Gln  Val Thr Val 
        995                 1000                 1005             


Gln Glu  Phe Ala Leu Phe Phe  Thr Ile Phe Asp Glu  Thr Lys Ser 
    1010                 1015                 1020             


Trp Tyr  Phe Thr Glu Asn Val  Glu Arg Asn Cys Arg  Ala Pro Cys 
    1025                 1030                 1035             


His Leu  Gln Met Glu Asp Pro  Thr Leu Lys Glu Asn  Tyr Arg Phe 
    1040                 1045                 1050             


His Ala  Ile Asn Gly Tyr Val  Met Asp Thr Leu Pro  Gly Leu Val 
    1055                 1060                 1065             


Met Ala  Gln Asn Gln Arg Ile  Arg Trp Tyr Leu Leu  Ser Met Gly 
    1070                 1075                 1080             


Ser Asn  Glu Asn Ile His Ser  Ile His Phe Ser Gly  His Val Phe 
    1085                 1090                 1095             


Ser Val  Arg Lys Lys Glu Glu  Tyr Lys Met Ala Val  Tyr Asn Leu 
    1100                 1105                 1110             


Tyr Pro  Gly Val Phe Glu Thr  Val Glu Met Leu Pro  Ser Lys Val 
    1115                 1120                 1125             


Gly Ile  Trp Arg Ile Glu Cys  Leu Ile Gly Glu His  Leu Gln Ala 
    1130                 1135                 1140             


Gly Met  Ser Thr Thr Phe Leu  Val Tyr Ser Lys Lys  Cys Gln Thr 
    1145                 1150                 1155             


Pro Leu  Gly Met Ala Ser Gly  His Ile Arg Asp Phe  Gln Ile Thr 
    1160                 1165                 1170             


Ala Ser  Gly Gln Tyr Gly Gln  Trp Ala Pro Lys Leu  Ala Arg Leu 
    1175                 1180                 1185             


His Tyr  Ser Gly Ser Ile Asn  Ala Trp Ser Thr Lys  Glu Pro Phe 
    1190                 1195                 1200             


Ser Trp  Ile Lys Val Asp Leu  Leu Ala Pro Met Ile  Ile His Gly 
    1205                 1210                 1215             


Ile Lys  Thr Gln Gly Ala Arg  Gln Lys Phe Ser Ser  Leu Tyr Ile 
    1220                 1225                 1230             


Ser Gln  Phe Ile Ile Met Tyr  Ser Leu Asp Gly Lys  Lys Trp Gln 
    1235                 1240                 1245             


Thr Tyr  Arg Gly Asn Ser Thr  Gly Thr Leu Met Val  Phe Phe Gly 
    1250                 1255                 1260             


Asn Val  Asp Ser Ser Gly Ile  Lys His Asn Ile Phe  Asn Pro Pro 
    1265                 1270                 1275             


Ile Ile  Ala Arg Tyr Ile Arg  Leu His Pro Thr His  Tyr Ser Ile 
    1280                 1285                 1290             


Arg Ser  Thr Leu Arg Met Glu  Leu Met Gly Cys Asp  Leu Asn Ser 
    1295                 1300                 1305             


Cys Ser  Met Pro Leu Gly Met  Glu Ser Lys Ala Ile  Ser Asp Ala 
    1310                 1315                 1320             


Gln Ile  Thr Ala Ser Ser Tyr  Phe Thr Asn Met Phe  Ala Thr Trp 
    1325                 1330                 1335             


Ser Pro  Ser Lys Ala Arg Leu  His Leu Gln Gly Arg  Ser Asn Ala 
    1340                 1345                 1350             


Trp Arg  Pro Gln Val Asn Asn  Pro Lys Glu Trp Leu  Gln Val Asp 
    1355                 1360                 1365             


Phe Gln  Lys Thr Met Lys Val  Thr Gly Val Thr Thr  Gln Gly Val 
    1370                 1375                 1380             


Lys Ser  Leu Leu Thr Ser Met  Tyr Val Lys Glu Phe  Leu Ile Ser 
    1385                 1390                 1395             


Ser Ser  Gln Asp Gly His Gln  Trp Thr Leu Phe Phe  Gln Asn Gly 
    1400                 1405                 1410             


Lys Val  Lys Val Phe Gln Gly  Asn Gln Asp Ser Phe  Thr Pro Val 
    1415                 1420                 1425             


Val Asn  Ser Leu Asp Pro Pro  Leu Leu Thr Arg Tyr  Leu Arg Ile 
    1430                 1435                 1440             


His Pro  Gln Ser Trp Val His  Gln Ile Ala Leu Arg  Met Glu Val 
    1445                 1450                 1455             


Leu Gly  Cys Glu Ala Gln Asp  Leu Tyr 
    1460                 1465         


<210>  6
<211>  579
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Fok I wildtype

<400>  6

Met Val Ser Lys Ile Arg Thr Phe Gly Trp Val Gln Asn Pro Gly Lys 
1               5                   10                  15      


Phe Glu Asn Leu Lys Arg Val Val Gln Val Phe Asp Arg Asn Ser Lys 
            20                  25                  30          


Val His Asn Glu Val Lys Asn Ile Lys Ile Pro Thr Leu Val Lys Glu 
        35                  40                  45              


Ser Lys Ile Gln Lys Glu Leu Val Ala Ile Met Asn Gln His Asp Leu 
    50                  55                  60                  


Ile Tyr Thr Tyr Lys Glu Leu Val Gly Thr Gly Thr Ser Ile Arg Ser 
65                  70                  75                  80  


Glu Ala Pro Cys Asp Ala Ile Ile Gln Ala Thr Ile Ala Asp Gln Gly 
                85                  90                  95      


Asn Lys Lys Gly Tyr Ile Asp Asn Trp Ser Ser Asp Gly Phe Leu Arg 
            100                 105                 110         


Trp Ala His Ala Leu Gly Phe Ile Glu Tyr Ile Asn Lys Ser Asp Ser 
        115                 120                 125             


Phe Val Ile Thr Asp Val Gly Leu Ala Tyr Ser Lys Ser Ala Asp Gly 
    130                 135                 140                 


Ser Ala Ile Glu Lys Glu Ile Leu Ile Glu Ala Ile Ser Ser Tyr Pro 
145                 150                 155                 160 


Pro Ala Ile Arg Ile Leu Thr Leu Leu Glu Asp Gly Gln His Leu Thr 
                165                 170                 175     


Lys Phe Asp Leu Gly Lys Asn Leu Gly Phe Ser Gly Glu Ser Gly Phe 
            180                 185                 190         


Thr Ser Leu Pro Glu Gly Ile Leu Leu Asp Thr Leu Ala Asn Ala Met 
        195                 200                 205             


Pro Lys Asp Lys Gly Glu Ile Arg Asn Asn Trp Glu Gly Ser Ser Asp 
    210                 215                 220                 


Lys Tyr Ala Arg Met Ile Gly Gly Trp Leu Asp Lys Leu Gly Leu Val 
225                 230                 235                 240 


Lys Gln Gly Lys Lys Glu Phe Ile Ile Pro Thr Leu Gly Lys Pro Asp 
                245                 250                 255     


Asn Lys Glu Phe Ile Ser His Ala Phe Lys Ile Thr Gly Glu Gly Leu 
            260                 265                 270         


Lys Val Leu Arg Arg Ala Lys Gly Ser Thr Lys Phe Thr Arg Val Pro 
        275                 280                 285             


Lys Arg Val Tyr Trp Glu Met Leu Ala Thr Asn Leu Thr Asp Lys Glu 
    290                 295                 300                 


Tyr Val Arg Thr Arg Arg Ala Leu Ile Leu Glu Ile Leu Ile Lys Ala 
305                 310                 315                 320 


Gly Ser Leu Lys Ile Glu Gln Ile Gln Asp Asn Leu Lys Lys Leu Gly 
                325                 330                 335     


Phe Asp Glu Val Ile Glu Thr Ile Glu Asn Asp Ile Lys Gly Leu Ile 
            340                 345                 350         


Asn Thr Gly Ile Phe Ile Glu Ile Lys Gly Arg Phe Tyr Gln Leu Lys 
        355                 360                 365             


Asp His Ile Leu Gln Phe Val Ile Pro Asn Arg Gly Val Thr Lys Gln 
    370                 375                 380                 


Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys 
385                 390                 395                 400 


Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg 
                405                 410                 415     


Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe 
            420                 425                 430         


Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys 
        435                 440                 445             


Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val 
    450                 455                 460                 


Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly 
465                 470                 475                 480 


Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn 
                485                 490                 495     


Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val 
            500                 505                 510         


Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr 
        515                 520                 525             


Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala 
    530                 535                 540                 


Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala 
545                 550                 555                 560 


Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu 
                565                 570                 575     


Ile Asn Phe 
            


<210>  7
<211>  195
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  OH-ZFN-right homology arm

(catalytic domain)

<400>  7

Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
1               5                   10                  15      


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
            20                  25                  30          


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
        35                  40                  45              


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
    50                  55                  60                  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
65                  70                  75                  80  


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
                85                  90                  95      


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Lys Glu Asn Gln Thr Arg 
            100                 105                 110         


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
        115                 120                 125             


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
    130                 135                 140                 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Lys Thr Asn Cys Asn Gly 
145                 150                 155                 160 


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
                165                 170                 175     


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
            180                 185                 190         


Glu Ile Asn 
        195 


<210>  8
<211>  196
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  OH-ZFN-left homology arm

(catalytic domain)

<400>  8

Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
1               5                   10                  15      


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
            20                  25                  30          


Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
        35                  40                  45              


Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg 
    50                  55                  60                  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
65                  70                  75                  80  


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
                85                  90                  95      


Gly Gln Ala Asp Glu Met Glu Arg Tyr Val Glu Glu Asn Gln Thr Arg 
            100                 105                 110         


Asn Lys His Leu Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
        115                 120                 125             


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
    130                 135                 140                 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
145                 150                 155                 160 


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
                165                 170                 175     


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
            180                 185                 190         


Glu Ile Asn Phe 
        195     


<210>  9
<211>  196
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Sharkey

(catalytic domain)

<400>  9

Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
1               5                   10                  15      


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
            20                  25                  30          


Arg Asn Pro Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
        35                  40                  45              


Phe Met Lys Val Tyr Gly Tyr Arg Gly Glu His Leu Gly Gly Ser Arg 
    50                  55                  60                  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
65                  70                  75                  80  


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
                85                  90                  95      


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
            100                 105                 110         


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
        115                 120                 125             


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
    130                 135                 140                 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
145                 150                 155                 160 


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
                165                 170                 175     


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
            180                 185                 190         


Glu Ile Asn Phe 
        195     


<210>  10
<211>  196
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Enhanced sharkey

FokI right monomer

(catalytic domain)

<400>  10

Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
1               5                   10                  15      


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
            20                  25                  30          


Arg Asn Pro Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
        35                  40                  45              


Phe Met Lys Val Tyr Gly Tyr Arg Gly Glu His Leu Gly Gly Ser Arg 
    50                  55                  60                  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
65                  70                  75                  80  


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
                85                  90                  95      


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
            100                 105                 110         


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
        115                 120                 125             


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
    130                 135                 140                 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn Arg Ile Thr Asn Cys Asn Gly 
145                 150                 155                 160 


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
                165                 170                 175     


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
            180                 185                 190         


Glu Ile Asn Phe 
        195     


<210>  11
<211>  196
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Enhanced sharkey left

FokI monomer 

(catalytic domain)

<400>  11

Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
1               5                   10                  15      


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
            20                  25                  30          


Arg Asn Pro Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
        35                  40                  45              


Phe Met Lys Val Tyr Gly Tyr Arg Gly Glu His Leu Gly Gly Ser Arg 
    50                  55                  60                  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
65                  70                  75                  80  


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
                85                  90                  95      


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg 
            100                 105                 110         


Asp Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
        115                 120                 125             


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
    130                 135                 140                 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
145                 150                 155                 160 


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
                165                 170                 175     


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
            180                 185                 190         


Glu Ile Asn Phe 
        195     


<210>  12
<211>  195
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  OH-ZFN-right homology arm + S418P, K441E

(catalytic domain)

<400>  12

Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
1               5                   10                  15      


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
            20                  25                  30          


Arg Asn Pro Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
        35                  40                  45              


Phe Met Lys Val Tyr Gly Tyr Arg Gly Glu His Leu Gly Gly Ser Arg 
    50                  55                  60                  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
65                  70                  75                  80  


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
                85                  90                  95      


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Lys Glu Asn Gln Thr Arg 
            100                 105                 110         


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
        115                 120                 125             


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
    130                 135                 140                 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Lys Thr Asn Cys Asn Gly 
145                 150                 155                 160 


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
                165                 170                 175     


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
            180                 185                 190         


Glu Ile Asn 
        195 


<210>  13
<211>  196
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  OH-ZFN-left homology arm +

S418P, K441E (catalytic domain)

<400>  13

Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
1               5                   10                  15      


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
            20                  25                  30          


Arg Asn Pro Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
        35                  40                  45              


Phe Met Lys Val Tyr Gly Tyr Arg Gly Glu His Leu Gly Gly Ser Arg 
    50                  55                  60                  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
65                  70                  75                  80  


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
                85                  90                  95      


Gly Gln Ala Asp Glu Met Glu Arg Tyr Val Glu Glu Asn Gln Thr Arg 
            100                 105                 110         


Asn Lys His Leu Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
        115                 120                 125             


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
    130                 135                 140                 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
145                 150                 155                 160 


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
                165                 170                 175     


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
            180                 185                 190         


Glu Ile Asn Phe 
        195     


<210>  14
<211>  196
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  OH-ZFN-right homology arm + S418P, K441E and H537R

       (catalytic domain)

<400>  14

Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
1               5                   10                  15      


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
            20                  25                  30          


Arg Asn Pro Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
        35                  40                  45              


Phe Met Lys Val Tyr Gly Tyr Arg Gly Glu His Leu Gly Gly Ser Arg 
    50                  55                  60                  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
65                  70                  75                  80  


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
                85                  90                  95      


Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Lys Glu Asn Gln Thr Arg 
            100                 105                 110         


Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
        115                 120                 125             


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
    130                 135                 140                 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn Arg Lys Thr Asn Cys Asn Gly 
145                 150                 155                 160 


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
                165                 170                 175     


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
            180                 185                 190         


Glu Ile Asn Phe 
        195     


<210>  15
<211>  196
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  OH-ZFN-left homology arm +

       S418P, K441E and N496D 

(catalytic domain)

<400>  15

Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His 
1               5                   10                  15      


Lys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala 
            20                  25                  30          


Arg Asn Pro Thr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe 
        35                  40                  45              


Phe Met Lys Val Tyr Gly Tyr Arg Gly Glu His Leu Gly Gly Ser Arg 
    50                  55                  60                  


Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly 
65                  70                  75                  80  


Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile 
                85                  90                  95      


Gly Gln Ala Asp Glu Met Glu Arg Tyr Val Glu Glu Asn Gln Thr Arg 
            100                 105                 110         


Asp Lys His Leu Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser 
        115                 120                 125             


Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn 
    130                 135                 140                 


Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn Gly 
145                 150                 155                 160 


Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys 
                165                 170                 175     


Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly 
            180                 185                 190         


Glu Ile Asn Phe 
        195     


<210>  16
<211>  8
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  zinc finger protein (amino acid sequence motif)


<220>
<221>  MISC_FEATURE
<222>  (1)..(1)
<223>  Xaa represents in total 2 amino acids, which can be any amino 
       acid

<220>
<221>  MISC_FEATURE
<222>  (3)..(3)
<223>  Xaa represents in total 2 or 4 amino acids, which can be any 
       amino acid

<220>
<221>  MISC_FEATURE
<222>  (5)..(5)
<223>  Xaa represents in total 3, 4 or 5 amino acids, which can be any 
       amino acid

<220>
<221>  misc_feature
<222>  (7)..(7)
<223>  Xaa can be any naturally occurring amino acid

<400>  16

Xaa Cys Xaa Cys Xaa His Xaa His 
1               5               


<210>  17
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Control 1 forward

<400>  17
cctgccttaa acccagccag                                                   20


<210>  18
<211>  26
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Control 1 reverse

<400>  18
atgacctcat gctcttggcc ctcgta                                            26


<210>  19
<211>  21
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Control 2 forward

<400>  19
tcccctccca gaaagacctg c                                                 21


<210>  20
<211>  21
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Control 2 reverse

<400>  20
tcccctccca gaaagacctg c                                                 21


<210>  21
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  probe

<400>  21
tacctaacgc actcctgggt ga                                                22


<210>  22
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Left genome specific forward

<400>  22
ttcgggtcac ctctcactcc                                                   20


<210>  23
<211>  21
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Left vector specific reverse

<400>  23
gacgcgcgtg aggaagagtt c                                                 21


<210>  24
<211>  19
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  probe

<400>  24
aggcgcaccg tgggcttgt                                                    19


<210>  25
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Right vector specific forward

<400>  25
ctgtggaatg tgtgtcagtt ag                                                22


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

<220>
<223>  Right genome specific reverse

<400>  26
ggctccatcg taagcaaacc                                                   20


<210>  27
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Right probe

<400>  27
cgcctctgcc tctgagctat                                                   20


<210>  28
<211>  19
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  vector specific PCR forward

<400>  28
cgagatgacc gagtacaag                                                    19


<210>  29
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  vector specific reverse

<400>  29
gctcgtagaa ggggaggttg                                                   20


<210>  30
<211>  19
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  probe

<400>  30
tcaccgagct gcaagaact                                                    19


<210>  31
<211>  12
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  left ZFN half-site

<400>  31
atcctgtccc ta                                                           12


<210>  32
<211>  12
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  right ZFN half-site

<400>  32
accccacagt gg                                                           12


