                         SEQUENCE LISTING

<110>  Autolus Ltd
 
<120>  CELL

<130>  P112306PCT

<150>  GB 1707779.3
<151>  2017-05-15

<160>  21    

<170>  PatentIn version 3.5

<210>  1
<211>  1085
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  polypeptide comprising two chimeric antigen receptors (CAR)

<400>  1

Met Ser Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu 
1               5                   10                  15      


His Ala Ala Arg Pro Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu 
            20                  25                  30          


Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln 
        35                  40                  45              


Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr 
    50                  55                  60                  


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


Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile 
                85                  90                  95      


Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly 
            100                 105                 110         


Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr 
        115                 120                 125             


Lys Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 
    130                 135                 140                 


Ser Gly Gly Gly Gly Ser Glu Val Lys Leu Gln Glu Ser Gly Pro Gly 
145                 150                 155                 160 


Leu Val Ala Pro Ser Gln Ser Leu Ser Val Thr Cys Thr Val Ser Gly 
                165                 170                 175     


Val Ser Leu Pro Asp Tyr Gly Val Ser Trp Ile Arg Gln Pro Pro Arg 
            180                 185                 190         


Lys Gly Leu Glu Trp Leu Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr 
        195                 200                 205             


Tyr Asn Ser Ala Leu Lys Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser 
    210                 215                 220                 


Lys Ser Gln Val Phe Leu Lys Met Asn Ser Leu Gln Thr Asp Asp Thr 
225                 230                 235                 240 


Ala Ile Tyr Tyr Cys Ala Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala 
                245                 250                 255     


Met Asp Tyr Trp Gly Gln Gly Thr Ser Val Thr Val Ser Ser Asp Pro 
            260                 265                 270         


Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala 
        275                 280                 285             


Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly 
    290                 295                 300                 


Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Phe Trp 
305                 310                 315                 320 


Val Leu Val Val Val Gly Gly Val Leu Ala Cys Tyr Ser Leu Leu Val 
                325                 330                 335     


Thr Val Ala Phe Ile Ile Phe Trp Val Arg Arg Val Lys Phe Ser Arg 
            340                 345                 350         


Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn 
        355                 360                 365             


Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg 
    370                 375                 380                 


Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro 
385                 390                 395                 400 


Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala 
                405                 410                 415     


Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His 
            420                 425                 430         


Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp 
        435                 440                 445             


Ala Leu His Met Gln Ala Leu Pro Pro Arg Arg Ala Glu Gly Arg Gly 
    450                 455                 460                 


Ser Leu Leu Thr Cys Gly Asp Val Glu Glu Asn Pro Gly Pro Met Ala 
465                 470                 475                 480 


Val Pro Thr Gln Val Leu Gly Leu Leu Leu Leu Trp Leu Thr Asp Ala 
                485                 490                 495     


Arg Cys Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser 
            500                 505                 510         


Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Asp Ile Tyr 
        515                 520                 525             


Phe Asn Leu Val Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu 
    530                 535                 540                 


Leu Ile Tyr Asp Thr Asn Arg Leu Ala Asp Gly Val Pro Ser Arg Phe 
545                 550                 555                 560 


Ser Gly Ser Gly Ser Gly Thr Gln Tyr Thr Leu Thr Ile Ser Ser Leu 
                565                 570                 575     


Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln His Tyr Lys Asn Tyr 
            580                 585                 590         


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


Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 
    610                 615                 620                 


Gly Gly Ser Arg Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu 
625                 630                 635                 640 


Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe 
                645                 650                 655     


Thr Leu Ser Asn Tyr Gly Met His Trp Ile Arg Gln Ala Pro Gly Lys 
            660                 665                 670         


Gly Leu Glu Trp Val Ser Ser Ile Ser Leu Asn Gly Gly Ser Thr Tyr 
        675                 680                 685             


Tyr Arg Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala 
    690                 695                 700                 


Lys Ser Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr 
705                 710                 715                 720 


Ala Val Tyr Tyr Cys Ala Ala Gln Asp Ala Tyr Thr Gly Gly Tyr Phe 
                725                 730                 735     


Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Met Asp Pro 
            740                 745                 750         


Ala Thr Thr Thr Lys Pro Val Leu Arg Thr Pro Ser Pro Val His Pro 
        755                 760                 765             


Thr Gly Thr Ser Gln Pro Gln Arg Pro Glu Asp Cys Arg Pro Arg Gly 
    770                 775                 780                 


Ser Val Lys Gly Thr Gly Leu Asp Phe Ala Cys Asp Ile Tyr Trp Ala 
785                 790                 795                 800 


Pro Leu Ala Gly Ile Cys Val Ala Leu Leu Leu Ser Leu Ile Ile Thr 
                805                 810                 815     


Leu Ile Cys Tyr His Arg Ser Arg Lys Arg Val Cys Lys Leu Lys Leu 
            820                 825                 830         


Leu Gln Thr Ile Gly Lys Gly Glu Phe Gly Asp Val Met Leu Gly Asp 
        835                 840                 845             


Tyr Arg Gly Asn Lys Val Ala Val Lys Cys Ile Lys Asn Asp Ala Thr 
    850                 855                 860                 


Ala Gln Ala Phe Leu Ala Glu Ala Ser Val Met Thr Gln Leu Arg His 
865                 870                 875                 880 


Ser Asn Leu Val Gln Leu Leu Gly Val Ile Val Glu Glu Lys Gly Gly 
                885                 890                 895     


Leu Tyr Ile Val Thr Glu Tyr Met Ala Lys Gly Ser Leu Val Asp Tyr 
            900                 905                 910         


Leu Arg Ser Arg Gly Arg Ser Val Leu Gly Gly Asp Cys Leu Leu Lys 
        915                 920                 925             


Phe Ser Leu Asp Val Cys Glu Ala Met Glu Tyr Leu Glu Gly Asn Asn 
    930                 935                 940                 


Phe Val His Arg Asp Leu Ala Ala Arg Asn Val Leu Val Ser Glu Asp 
945                 950                 955                 960 


Asn Val Ala Lys Val Ser Asp Phe Gly Leu Thr Lys Glu Ala Ser Ser 
                965                 970                 975     


Thr Gln Asp Thr Gly Lys Leu Pro Val Lys Trp Thr Ala Pro Glu Ala 
            980                 985                 990         


Leu Arg Glu Lys Lys Phe Ser Thr  Lys Ser Asp Val Trp  Ser Phe Gly 
        995                 1000                 1005             


Ile Leu  Leu Trp Glu Ile Tyr  Ser Phe Gly Arg Val  Pro Tyr Pro 
    1010                 1015                 1020             


Arg Ile  Pro Leu Lys Asp Val  Val Pro Arg Val Glu  Lys Gly Tyr 
    1025                 1030                 1035             


Lys Met  Asp Ala Pro Asp Gly  Cys Pro Pro Ala Val  Tyr Glu Val 
    1040                 1045                 1050             


Met Lys  Asn Cys Trp His Leu  Asp Ala Ala Met Arg  Pro Ser Phe 
    1055                 1060                 1065             


Leu Gln  Leu Arg Glu Gln Leu  Glu His Ile Lys Thr  His Glu Leu 
    1070                 1075                 1080             


His Leu  
    1085 


<210>  2
<211>  1278
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  polypeptide comprising two CARs

<400>  2

Met Ser Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu 
1               5                   10                  15      


His Ala Ala Arg Pro Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu 
            20                  25                  30          


Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln 
        35                  40                  45              


Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr 
    50                  55                  60                  


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


Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile 
                85                  90                  95      


Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly 
            100                 105                 110         


Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr 
        115                 120                 125             


Lys Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 
    130                 135                 140                 


Ser Gly Gly Gly Gly Ser Glu Val Lys Leu Gln Glu Ser Gly Pro Gly 
145                 150                 155                 160 


Leu Val Ala Pro Ser Gln Ser Leu Ser Val Thr Cys Thr Val Ser Gly 
                165                 170                 175     


Val Ser Leu Pro Asp Tyr Gly Val Ser Trp Ile Arg Gln Pro Pro Arg 
            180                 185                 190         


Lys Gly Leu Glu Trp Leu Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr 
        195                 200                 205             


Tyr Asn Ser Ala Leu Lys Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser 
    210                 215                 220                 


Lys Ser Gln Val Phe Leu Lys Met Asn Ser Leu Gln Thr Asp Asp Thr 
225                 230                 235                 240 


Ala Ile Tyr Tyr Cys Ala Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala 
                245                 250                 255     


Met Asp Tyr Trp Gly Gln Gly Thr Ser Val Thr Val Ser Ser Asp Pro 
            260                 265                 270         


Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala 
        275                 280                 285             


Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly 
    290                 295                 300                 


Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Phe Trp 
305                 310                 315                 320 


Val Leu Val Val Val Gly Gly Val Leu Ala Cys Tyr Ser Leu Leu Val 
                325                 330                 335     


Thr Val Ala Phe Ile Ile Phe Trp Val Arg Arg Val Lys Phe Ser Arg 
            340                 345                 350         


Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn 
        355                 360                 365             


Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg 
    370                 375                 380                 


Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro 
385                 390                 395                 400 


Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala 
                405                 410                 415     


Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His 
            420                 425                 430         


Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp 
        435                 440                 445             


Ala Leu His Met Gln Ala Leu Pro Pro Arg Arg Ala Glu Gly Arg Gly 
    450                 455                 460                 


Ser Leu Leu Thr Cys Gly Asp Val Glu Glu Asn Pro Gly Pro Met Ala 
465                 470                 475                 480 


Val Pro Thr Gln Val Leu Gly Leu Leu Leu Leu Trp Leu Thr Asp Ala 
                485                 490                 495     


Arg Cys Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser 
            500                 505                 510         


Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Asp Ile Tyr 
        515                 520                 525             


Phe Asn Leu Val Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu 
    530                 535                 540                 


Leu Ile Tyr Asp Thr Asn Arg Leu Ala Asp Gly Val Pro Ser Arg Phe 
545                 550                 555                 560 


Ser Gly Ser Gly Ser Gly Thr Gln Tyr Thr Leu Thr Ile Ser Ser Leu 
                565                 570                 575     


Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln His Tyr Lys Asn Tyr 
            580                 585                 590         


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


Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 
    610                 615                 620                 


Gly Gly Ser Arg Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu 
625                 630                 635                 640 


Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe 
                645                 650                 655     


Thr Leu Ser Asn Tyr Gly Met His Trp Ile Arg Gln Ala Pro Gly Lys 
            660                 665                 670         


Gly Leu Glu Trp Val Ser Ser Ile Ser Leu Asn Gly Gly Ser Thr Tyr 
        675                 680                 685             


Tyr Arg Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala 
    690                 695                 700                 


Lys Ser Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr 
705                 710                 715                 720 


Ala Val Tyr Tyr Cys Ala Ala Gln Asp Ala Tyr Thr Gly Gly Tyr Phe 
                725                 730                 735     


Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Met Asp Pro 
            740                 745                 750         


Ala Thr Thr Thr Lys Pro Val Leu Arg Thr Pro Ser Pro Val His Pro 
        755                 760                 765             


Thr Gly Thr Ser Gln Pro Gln Arg Pro Glu Asp Cys Arg Pro Arg Gly 
    770                 775                 780                 


Ser Val Lys Gly Thr Gly Leu Asp Phe Ala Cys Asp Ile Tyr Trp Ala 
785                 790                 795                 800 


Pro Leu Ala Gly Ile Cys Val Ala Leu Leu Leu Ser Leu Ile Ile Thr 
                805                 810                 815     


Leu Ile Cys Tyr His Arg Ser Arg Lys Arg Val Cys Lys Ser Ala Ile 
            820                 825                 830         


Gln Ala Ala Trp Pro Ser Gly Thr Glu Cys Ile Ala Lys Tyr Asn Phe 
        835                 840                 845             


His Gly Thr Ala Glu Gln Asp Leu Pro Phe Cys Lys Gly Asp Val Leu 
    850                 855                 860                 


Thr Ile Val Ala Val Thr Lys Asp Pro Asn Trp Tyr Lys Ala Lys Asn 
865                 870                 875                 880 


Lys Val Gly Arg Glu Gly Ile Ile Pro Ala Asn Tyr Val Gln Lys Arg 
                885                 890                 895     


Glu Gly Val Lys Ala Gly Thr Lys Leu Ser Leu Met Pro Trp Phe His 
            900                 905                 910         


Gly Lys Ile Thr Arg Glu Gln Ala Glu Arg Leu Leu Tyr Pro Pro Glu 
        915                 920                 925             


Thr Gly Leu Phe Leu Val Arg Glu Ser Thr Asn Tyr Pro Gly Asp Tyr 
    930                 935                 940                 


Thr Leu Cys Val Ser Cys Asp Gly Lys Val Glu His Tyr Arg Ile Met 
945                 950                 955                 960 


Tyr His Ala Ser Lys Leu Ser Ile Asp Glu Glu Val Tyr Phe Glu Asn 
                965                 970                 975     


Leu Met Gln Leu Val Glu His Tyr Thr Ser Asp Ala Asp Gly Leu Cys 
            980                 985                 990         


Thr Arg Leu Ile Lys Pro Lys Val  Met Glu Gly Thr Val  Ala Ala Gln 
        995                 1000                 1005             


Asp Glu  Phe Tyr Arg Ser Gly  Trp Ala Leu Asn Met  Lys Glu Leu 
    1010                 1015                 1020             


Lys Leu  Leu Gln Thr Ile Gly  Lys Gly Glu Phe Gly  Asp Val Met 
    1025                 1030                 1035             


Leu Gly  Asp Tyr Arg Gly Asn  Lys Val Ala Val Lys  Cys Ile Lys 
    1040                 1045                 1050             


Asn Asp  Ala Thr Ala Gln Ala  Phe Leu Ala Glu Ala  Ser Val Met 
    1055                 1060                 1065             


Thr Gln  Leu Arg His Ser Asn  Leu Val Gln Leu Leu  Gly Val Ile 
    1070                 1075                 1080             


Val Glu  Glu Lys Gly Gly Leu  Tyr Ile Val Thr Glu  Tyr Met Ala 
    1085                 1090                 1095             


Lys Gly  Ser Leu Val Asp Tyr  Leu Arg Ser Arg Gly  Arg Ser Val 
    1100                 1105                 1110             


Leu Gly  Gly Asp Cys Leu Leu  Lys Phe Ser Leu Asp  Val Cys Glu 
    1115                 1120                 1125             


Ala Met  Glu Tyr Leu Glu Gly  Asn Asn Phe Val His  Arg Asp Leu 
    1130                 1135                 1140             


Ala Ala  Arg Asn Val Leu Val  Ser Glu Asp Asn Val  Ala Lys Val 
    1145                 1150                 1155             


Ser Asp  Phe Gly Leu Thr Lys  Glu Ala Ser Ser Thr  Gln Asp Thr 
    1160                 1165                 1170             


Gly Lys  Leu Pro Val Lys Trp  Thr Ala Pro Glu Ala  Leu Arg Glu 
    1175                 1180                 1185             


Lys Lys  Phe Ser Thr Lys Ser  Asp Val Trp Ser Phe  Gly Ile Leu 
    1190                 1195                 1200             


Leu Trp  Glu Ile Tyr Ser Phe  Gly Arg Val Pro Tyr  Pro Arg Ile 
    1205                 1210                 1215             


Pro Leu  Lys Asp Val Val Pro  Arg Val Glu Lys Gly  Tyr Lys Met 
    1220                 1225                 1230             


Asp Ala  Pro Asp Gly Cys Pro  Pro Ala Val Tyr Glu  Val Met Lys 
    1235                 1240                 1245             


Asn Cys  Trp His Leu Asp Ala  Ala Met Arg Pro Ser  Phe Leu Gln 
    1250                 1255                 1260             


Leu Arg  Glu Gln Leu Glu His  Ile Lys Thr His Glu  Leu His Leu 
    1265                 1270                 1275             


<210>  3
<211>  21
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  signal peptide

<400>  3

Met Gly Thr Ser Leu Leu Cys Trp Met Ala Leu Cys Leu Leu Gly Ala 
1               5                   10                  15      


Asp His Ala Asp Gly 
            20      


<210>  4
<211>  21
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  signal peptide derived from IgG1

<400>  4

Met Ser Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu 
1               5                   10                  15      


His Ala Ala Arg Pro 
            20      


<210>  5
<211>  20
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  signal peptide derived from CD8

<400>  5

Met Ala Val Pro Thr Gln Val Leu Gly Leu Leu Leu Leu Trp Leu Thr 
1               5                   10                  15      


Asp Ala Arg Cys 
            20  


<210>  6
<211>  234
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  spacer sequence, hinge-CH2CH3 of human IgG1

<400>  6

Ala Glu Pro Lys Ser Pro Asp Lys Thr His Thr Cys Pro Pro Cys Pro 
1               5                   10                  15      


Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro 
            20                  25                  30          


Lys Asp Thr Leu Met Ile Ala Arg Thr Pro Glu Val Thr Cys Val Val 
        35                  40                  45              


Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val 
    50                  55                  60                  


Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln 
65                  70                  75                  80  


Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln 
                85                  90                  95      


Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala 
            100                 105                 110         


Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro 
        115                 120                 125             


Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr 
    130                 135                 140                 


Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser 
145                 150                 155                 160 


Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr 
                165                 170                 175     


Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr 
            180                 185                 190         


Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe 
        195                 200                 205             


Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys 
    210                 215                 220                 


Ser Leu Ser Leu Ser Pro Gly Lys Lys Asp 
225                 230                 


<210>  7
<211>  46
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  spacer sequence, human CD8 stalk

<400>  7

Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala 
1               5                   10                  15      


Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly 
            20                  25                  30          


Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile 
        35                  40                  45      


<210>  8
<211>  44
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  spacer sequence, mouse CD8a stalk

<400>  8

Ala Thr Thr Thr Lys Pro Val Leu Arg Thr Pro Ser Pro Val His Pro 
1               5                   10                  15      


Thr Gly Thr Ser Gln Pro Gln Arg Pro Glu Asp Cys Arg Pro Arg Gly 
            20                  25                  30          


Ser Val Lys Gly Thr Gly Leu Asp Phe Ala Cys Asp 
        35                  40                  


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

<220>
<223>  spacer sequence, human IgG1 hinge

<400>  9

Ala Glu Pro Lys Ser Pro Asp Lys Thr His Thr Cys Pro Pro Cys Pro 
1               5                   10                  15      


Lys Asp Pro Lys 
            20  


<210>  10
<211>  185
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  spacer sequence, CD2 ectodomain

<400>  10

Lys Glu Ile Thr Asn Ala Leu Glu Thr Trp Gly Ala Leu Gly Gln Asp 
1               5                   10                  15      


Ile Asn Leu Asp Ile Pro Ser Phe Gln Met Ser Asp Asp Ile Asp Asp 
            20                  25                  30          


Ile Lys Trp Glu Lys Thr Ser Asp Lys Lys Lys Ile Ala Gln Phe Arg 
        35                  40                  45              


Lys Glu Lys Glu Thr Phe Lys Glu Lys Asp Thr Tyr Lys Leu Phe Lys 
    50                  55                  60                  


Asn Gly Thr Leu Lys Ile Lys His Leu Lys Thr Asp Asp Gln Asp Ile 
65                  70                  75                  80  


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


Ile Phe Asp Leu Lys Ile Gln Glu Arg Val Ser Lys Pro Lys Ile Ser 
            100                 105                 110         


Trp Thr Cys Ile Asn Thr Thr Leu Thr Cys Glu Val Met Asn Gly Thr 
        115                 120                 125             


Asp Pro Glu Leu Asn Leu Tyr Gln Asp Gly Lys His Leu Lys Leu Ser 
    130                 135                 140                 


Gln Arg Val Ile Thr His Lys Trp Thr Thr Ser Leu Ser Ala Lys Phe 
145                 150                 155                 160 


Lys Cys Thr Ala Gly Asn Lys Val Ser Lys Glu Ser Ser Val Glu Pro 
                165                 170                 175     


Val Ser Cys Pro Glu Lys Gly Leu Asp 
            180                 185 


<210>  11
<211>  259
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  spacer sequence, CD34 ectodomain

<400>  11

Ser Leu Asp Asn Asn Gly Thr Ala Thr Pro Glu Leu Pro Thr Gln Gly 
1               5                   10                  15      


Thr Phe Ser Asn Val Ser Thr Asn Val Ser Tyr Gln Glu Thr Thr Thr 
            20                  25                  30          


Pro Ser Thr Leu Gly Ser Thr Ser Leu His Pro Val Ser Gln His Gly 
        35                  40                  45              


Asn Glu Ala Thr Thr Asn Ile Thr Glu Thr Thr Val Lys Phe Thr Ser 
    50                  55                  60                  


Thr Ser Val Ile Thr Ser Val Tyr Gly Asn Thr Asn Ser Ser Val Gln 
65                  70                  75                  80  


Ser Gln Thr Ser Val Ile Ser Thr Val Phe Thr Thr Pro Ala Asn Val 
                85                  90                  95      


Ser Thr Pro Glu Thr Thr Leu Lys Pro Ser Leu Ser Pro Gly Asn Val 
            100                 105                 110         


Ser Asp Leu Ser Thr Thr Ser Thr Ser Leu Ala Thr Ser Pro Thr Lys 
        115                 120                 125             


Pro Tyr Thr Ser Ser Ser Pro Ile Leu Ser Asp Ile Lys Ala Glu Ile 
    130                 135                 140                 


Lys Cys Ser Gly Ile Arg Glu Val Lys Leu Thr Gln Gly Ile Cys Leu 
145                 150                 155                 160 


Glu Gln Asn Lys Thr Ser Ser Cys Ala Glu Phe Lys Lys Asp Arg Gly 
                165                 170                 175     


Glu Gly Leu Ala Arg Val Leu Cys Gly Glu Glu Gln Ala Asp Ala Asp 
            180                 185                 190         


Ala Gly Ala Gln Val Cys Ser Leu Leu Leu Ala Gln Ser Glu Val Arg 
        195                 200                 205             


Pro Gln Cys Leu Leu Leu Val Leu Ala Asn Arg Thr Glu Ile Ser Ser 
    210                 215                 220                 


Lys Leu Gln Leu Met Lys Lys His Gln Ser Asp Leu Lys Lys Leu Gly 
225                 230                 235                 240 


Ile Leu Asp Phe Thr Glu Gln Asp Val Ala Ser His Gln Ser Tyr Ser 
                245                 250                 255     


Gln Lys Thr 
            


<210>  12
<211>  140
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  sequence comprising CD28 transmembrane domain and CD3 Z 
       endodomain

<400>  12

Phe Trp Val Leu Val Val Val Gly Gly Val Leu Ala Cys Tyr Ser Leu 
1               5                   10                  15      


Leu Val Thr Val Ala Phe Ile Ile Phe Trp Val Arg Arg Val Lys Phe 
            20                  25                  30          


Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu 
        35                  40                  45              


Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp 
    50                  55                  60                  


Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys 
65                  70                  75                  80  


Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala 
                85                  90                  95      


Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys 
            100                 105                 110         


Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr 
        115                 120                 125             


Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg 
    130                 135                 140 


<210>  13
<211>  180
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  sequence comprising CD28 transmembrane domain and CD28 and CD3 
       Zeta endodomains

<400>  13

Phe Trp Val Leu Val Val Val Gly Gly Val Leu Ala Cys Tyr Ser Leu 
1               5                   10                  15      


Leu Val Thr Val Ala Phe Ile Ile Phe Trp Val Arg Ser Lys Arg Ser 
            20                  25                  30          


Arg Leu Leu His Ser Asp Tyr Met Asn Met Thr Pro Arg Arg Pro Gly 
        35                  40                  45              


Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala Pro Pro Arg Asp Phe Ala 
    50                  55                  60                  


Ala Tyr Arg Ser Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala 
65                  70                  75                  80  


Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg 
                85                  90                  95      


Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu 
            100                 105                 110         


Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn 
        115                 120                 125             


Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met 
    130                 135                 140                 


Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly 
145                 150                 155                 160 


Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala 
                165                 170                 175     


Leu Pro Pro Arg 
            180 


<210>  14
<211>  216
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  sequence comprising CD28 transmembrane domain and CD28, OX40 and 
       CD3 Zeta endodomains

<400>  14

Phe Trp Val Leu Val Val Val Gly Gly Val Leu Ala Cys Tyr Ser Leu 
1               5                   10                  15      


Leu Val Thr Val Ala Phe Ile Ile Phe Trp Val Arg Ser Lys Arg Ser 
            20                  25                  30          


Arg Leu Leu His Ser Asp Tyr Met Asn Met Thr Pro Arg Arg Pro Gly 
        35                  40                  45              


Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala Pro Pro Arg Asp Phe Ala 
    50                  55                  60                  


Ala Tyr Arg Ser Arg Asp Gln Arg Leu Pro Pro Asp Ala His Lys Pro 
65                  70                  75                  80  


Pro Gly Gly Gly Ser Phe Arg Thr Pro Ile Gln Glu Glu Gln Ala Asp 
                85                  90                  95      


Ala His Ser Thr Leu Ala Lys Ile Arg Val Lys Phe Ser Arg Ser Ala 
            100                 105                 110         


Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu 
        115                 120                 125             


Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly 
    130                 135                 140                 


Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu 
145                 150                 155                 160 


Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser 
                165                 170                 175     


Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly 
            180                 185                 190         


Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu 
        195                 200                 205             


His Met Gln Ala Leu Pro Pro Arg 
    210                 215     


<210>  15
<211>  449
<212>  PRT
<213>  Homo sapiens


<220>
<221>  MISC_FEATURE
<223>  full length C-terminal Src kinase (CSK)

<400>  15

Ser Ala Ile Gln Ala Ala Trp Pro Ser Gly Thr Glu Cys Ile Ala Lys 
1               5                   10                  15      


Tyr Asn Phe His Gly Thr Ala Glu Gln Asp Leu Pro Phe Cys Lys Gly 
            20                  25                  30          


Asp Val Leu Thr Ile Val Ala Val Thr Lys Asp Pro Asn Trp Tyr Lys 
        35                  40                  45              


Ala Lys Asn Lys Val Gly Arg Glu Gly Ile Ile Pro Ala Asn Tyr Val 
    50                  55                  60                  


Gln Lys Arg Glu Gly Val Lys Ala Gly Thr Lys Leu Ser Leu Met Pro 
65                  70                  75                  80  


Trp Phe His Gly Lys Ile Thr Arg Glu Gln Ala Glu Arg Leu Leu Tyr 
                85                  90                  95      


Pro Pro Glu Thr Gly Leu Phe Leu Val Arg Glu Ser Thr Asn Tyr Pro 
            100                 105                 110         


Gly Asp Tyr Thr Leu Cys Val Ser Cys Asp Gly Lys Val Glu His Tyr 
        115                 120                 125             


Arg Ile Met Tyr His Ala Ser Lys Leu Ser Ile Asp Glu Glu Val Tyr 
    130                 135                 140                 


Phe Glu Asn Leu Met Gln Leu Val Glu His Tyr Thr Ser Asp Ala Asp 
145                 150                 155                 160 


Gly Leu Cys Thr Arg Leu Ile Lys Pro Lys Val Met Glu Gly Thr Val 
                165                 170                 175     


Ala Ala Gln Asp Glu Phe Tyr Arg Ser Gly Trp Ala Leu Asn Met Lys 
            180                 185                 190         


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


Met Leu Gly Asp Tyr Arg Gly Asn Lys Val Ala Val Lys Cys Ile Lys 
    210                 215                 220                 


Asn Asp Ala Thr Ala Gln Ala Phe Leu Ala Glu Ala Ser Val Met Thr 
225                 230                 235                 240 


Gln Leu Arg His Ser Asn Leu Val Gln Leu Leu Gly Val Ile Val Glu 
                245                 250                 255     


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


Leu Val Asp Tyr Leu Arg Ser Arg Gly Arg Ser Val Leu Gly Gly Asp 
        275                 280                 285             


Cys Leu Leu Lys Phe Ser Leu Asp Val Cys Glu Ala Met Glu Tyr Leu 
    290                 295                 300                 


Glu Gly Asn Asn Phe Val His Arg Asp Leu Ala Ala Arg Asn Val Leu 
305                 310                 315                 320 


Val Ser Glu Asp Asn Val Ala Lys Val Ser Asp Phe Gly Leu Thr Lys 
                325                 330                 335     


Glu Ala Ser Ser Thr Gln Asp Thr Gly Lys Leu Pro Val Lys Trp Thr 
            340                 345                 350         


Ala Pro Glu Ala Leu Arg Glu Lys Lys Phe Ser Thr Lys Ser Asp Val 
        355                 360                 365             


Trp Ser Phe Gly Ile Leu Leu Trp Glu Ile Tyr Ser Phe Gly Arg Val 
    370                 375                 380                 


Pro Tyr Pro Arg Ile Pro Leu Lys Asp Val Val Pro Arg Val Glu Lys 
385                 390                 395                 400 


Gly Tyr Lys Met Asp Ala Pro Asp Gly Cys Pro Pro Ala Val Tyr Glu 
                405                 410                 415     


Val Met Lys Asn Cys Trp His Leu Asp Ala Ala Met Arg Pro Ser Phe 
            420                 425                 430         


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


Leu 
    


<210>  16
<211>  256
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  sequence of tyrosine kinase domain of CSK

<400>  16

Leu Lys Leu Leu Gln Thr Ile Gly Lys Gly Glu Phe Gly Asp Val Met 
1               5                   10                  15      


Leu Gly Asp Tyr Arg Gly Asn Lys Val Ala Val Lys Cys Ile Lys Asn 
            20                  25                  30          


Asp Ala Thr Ala Gln Ala Phe Leu Ala Glu Ala Ser Val Met Thr Gln 
        35                  40                  45              


Leu Arg His Ser Asn Leu Val Gln Leu Leu Gly Val Ile Val Glu Glu 
    50                  55                  60                  


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


Val Asp Tyr Leu Arg Ser Arg Gly Arg Ser Val Leu Gly Gly Asp Cys 
                85                  90                  95      


Leu Leu Lys Phe Ser Leu Asp Val Cys Glu Ala Met Glu Tyr Leu Glu 
            100                 105                 110         


Gly Asn Asn Phe Val His Arg Asp Leu Ala Ala Arg Asn Val Leu Val 
        115                 120                 125             


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


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


Pro Glu Ala Leu Arg Glu Lys Lys Phe Ser Thr Lys Ser Asp Val Trp 
                165                 170                 175     


Ser Phe Gly Ile Leu Leu Trp Glu Ile Tyr Ser Phe Gly Arg Val Pro 
            180                 185                 190         


Tyr Pro Arg Ile Pro Leu Lys Asp Val Val Pro Arg Val Glu Lys Gly 
        195                 200                 205             


Tyr Lys Met Asp Ala Pro Asp Gly Cys Pro Pro Ala Val Tyr Glu Val 
    210                 215                 220                 


Met Lys Asn Cys Trp His Leu Asp Ala Ala Met Arg Pro Ser Phe Leu 
225                 230                 235                 240 


Gln Leu Arg Glu Gln Leu Glu His Ile Lys Thr His Glu Leu His Leu 
                245                 250                 255     


<210>  17
<211>  20
<212>  PRT
<213>  Foot-and-mouth disease virus

<400>  17

Arg Ala Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly Asp Val Glu Glu 
1               5                   10                  15      


Asn Pro Gly Pro 
            20  


<210>  18
<211>  3255
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  dual CAR system using tyrosine kinase domain CSK as endodomain 
       (CD19 and CD33)

<400>  18
atgagcctgc ccgtgaccgc cctgctgctg cccctggccc tgctgctgca cgccgccaga       60

ccagacatcc agatgaccca gaccaccagc agcctgagcg ccagcctggg cgaccgggtg      120

accatcagct gcagagccag ccaggacatc agcaagtacc tgaactggta ccagcagaag      180

cccgacggca ccgtgaagct gctgatctac cacaccagcc ggctgcacag cggcgtgccc      240

agccggttca gcggcagcgg cagcggcacc gactacagcc tgaccatcag caacctggag      300

caggaggaca tcgccaccta cttctgccag cagggcaaca ccctgcccta caccttcgga      360

ggcggcacca agctggagat caccaaggcc ggaggcggag gctctggcgg aggcggctct      420

ggcggaggcg gctctggcgg aggcggcagc gaggtgaagc tgcaggagtc tggcccaggc      480

ctggtggccc caagccagag cctgagcgtg acctgcaccg tgagcggcgt gagcctgccc      540

gactacggcg tgagctggat caggcagccc ccacggaagg gcctggagtg gctgggcgtg      600

atctggggca gcgagaccac ctactacaac agcgccctga agagccggct gaccatcatc      660

aaggacaaca gcaagagcca ggtgttcctg aagatgaaca gcctgcagac cgacgacacc      720

gccatctact actgcgccaa gcactactac tatggcggca gctacgctat ggactactgg      780

ggccagggca ccagcgtgac cgtgagctca gatcccacca cgacgccagc gccgcgacca      840

ccaacaccgg cgcccaccat cgcgtcgcag cccctgtccc tgcgcccaga ggcgtgccgg      900

ccagcggcgg ggggcgcagt gcacacgagg gggctggact tcgcctgtga tatcttttgg      960

gtgctggtgg tggttggtgg agtcctggct tgctatagct tgctagtaac agtggccttt     1020

attattttct gggtgaggag agtgaagttc agcaggagcg cagacgcccc cgcgtaccag     1080

cagggccaga accagctcta taacgagctc aatctaggac gaagagagga gtacgatgtt     1140

ttggacaaga gacgtggccg ggaccctgag atggggggaa agccgagaag gaagaaccct     1200

caggaaggcc tgtacaatga actgcagaaa gataagatgg cggaggccta cagtgagatt     1260

gggatgaaag gcgagcgccg gaggggcaag gggcacgatg gcctttacca gggtctcagt     1320

acagccacca aggacaccta cgacgccctt cacatgcagg ccctgcctcc tcgcagagcc     1380

gagggcaggg gaagtcttct aacatgcggg gacgtggagg aaaatcccgg gcccatggcc     1440

gtgcccactc aggtcctggg gttgttgcta ctgtggctta cagatgccag atgtgacatc     1500

cagatgacac agtctccatc ttccctgtct gcatctgtcg gagatcgcgt caccatcacc     1560

tgtcgagcaa gtgaggacat ttattttaat ttagtgtggt atcagcagaa accaggaaag     1620

gcccctaagc tcctgatcta tgatacaaat cgcttggcag atggggtccc atcacggttc     1680

agtggctctg gatctggcac acagtatact ctaaccataa gtagcctgca acccgaagat     1740

ttcgcaacct attattgtca acactataag aattatccgc tcacgttcgg tcaggggacc     1800

aagctggaaa tcaaaagatc tggtggcgga gggtcaggag gcggaggcag cggaggcggt     1860

ggctcgggag gcggaggctc gagatctgag gtgcagttgg tggagtctgg gggcggcttg     1920

gtgcagcctg gagggtccct gaggctctcc tgtgcagcct caggattcac tctcagtaat     1980

tatggcatgc actggatcag gcaggctcca gggaagggtc tggagtgggt ctcgtctatt     2040

agtcttaatg gtggtagcac ttactatcga gactccgtga agggccgatt cactatctcc     2100

agggacaatg caaaaagcac cctctacctt caaatgaata gtctgagggc cgaggacacg     2160

gccgtctatt actgtgcagc acaggacgct tatacgggag gttactttga ttactggggc     2220

caaggaacgc tggtcacagt ctcgtctatg gatcccgcca ccacaaccaa gcccgtgctg     2280

cggaccccaa gccctgtgca ccctaccggc accagccagc ctcagagacc cgaggactgc     2340

cggcctcggg gcagcgtgaa gggcaccggc ctggacttcg cctgcgacat ctactgggca     2400

cctctggccg gaatatgcgt ggcactgctg ctgagcctca tcatcaccct gatctgttat     2460

caccgaagcc gcaagcgggt gtgtaaactg aagctgctgc agaccatcgg caagggcgag     2520

tttggagatg tgatgctggg cgactaccgg ggcaacaagg tggcagtgaa gtgcatcaag     2580

aacgacgcta cagcccaggc ttttctggcc gaagccagcg tgatgaccca gctgagacac     2640

agcaatctgg tgcagctgct gggcgtgatc gtggaagaaa aaggcggcct gtatatcgtg     2700

accgagtaca tggccaaggg cagcctggtg gactacctga gaagtagagg cagaagcgtg     2760

ctcggaggcg actgcctgct gaagtttagc ctggatgtgt gcgaggctat ggaatacctg     2820

gaaggcaaca acttcgtgca ccgcgatctg gccgccagaa atgtgctggt gtccgaggac     2880

aacgtggcca aggtgtccga tttcggcctg accaaagagg ccagcagcac ccaggataca     2940

ggcaagctgc ccgtgaaatg gacagcccct gaggctctga gagagaagaa gttcagcacc     3000

aagagcgacg tgtggtcctt cggcatcctg ctgtgggaaa tctacagctt cggcagagtg     3060

ccctatccta gaatccctct gaaggacgtg gtgcccagag tggaaaaggg ctacaagatg     3120

gatgcccctg acggctgtcc tcctgccgtg tacgaagtga tgaagaactg ctggcacctg     3180

gacgccgcta tgaggccatc tttcctgcag ctgagagagc agctggaaca catcaagacc     3240

cacgagctgc acctg                                                      3255


<210>  19
<211>  3834
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  dual CAR system using full sequence CSK as endodomain (CD19 and 
       CD33)

<400>  19
atgagcctgc ccgtgaccgc cctgctgctg cccctggccc tgctgctgca cgccgccaga       60

ccagacatcc agatgaccca gaccaccagc agcctgagcg ccagcctggg cgaccgggtg      120

accatcagct gcagagccag ccaggacatc agcaagtacc tgaactggta ccagcagaag      180

cccgacggca ccgtgaagct gctgatctac cacaccagcc ggctgcacag cggcgtgccc      240

agccggttca gcggcagcgg cagcggcacc gactacagcc tgaccatcag caacctggag      300

caggaggaca tcgccaccta cttctgccag cagggcaaca ccctgcccta caccttcgga      360

ggcggcacca agctggagat caccaaggcc ggaggcggag gctctggcgg aggcggctct      420

ggcggaggcg gctctggcgg aggcggcagc gaggtgaagc tgcaggagtc tggcccaggc      480

ctggtggccc caagccagag cctgagcgtg acctgcaccg tgagcggcgt gagcctgccc      540

gactacggcg tgagctggat caggcagccc ccacggaagg gcctggagtg gctgggcgtg      600

atctggggca gcgagaccac ctactacaac agcgccctga agagccggct gaccatcatc      660

aaggacaaca gcaagagcca ggtgttcctg aagatgaaca gcctgcagac cgacgacacc      720

gccatctact actgcgccaa gcactactac tatggcggca gctacgctat ggactactgg      780

ggccagggca ccagcgtgac cgtgagctca gatcccacca cgacgccagc gccgcgacca      840

ccaacaccgg cgcccaccat cgcgtcgcag cccctgtccc tgcgcccaga ggcgtgccgg      900

ccagcggcgg ggggcgcagt gcacacgagg gggctggact tcgcctgtga tatcttttgg      960

gtgctggtgg tggttggtgg agtcctggct tgctatagct tgctagtaac agtggccttt     1020

attattttct gggtgaggag agtgaagttc agcaggagcg cagacgcccc cgcgtaccag     1080

cagggccaga accagctcta taacgagctc aatctaggac gaagagagga gtacgatgtt     1140

ttggacaaga gacgtggccg ggaccctgag atggggggaa agccgagaag gaagaaccct     1200

caggaaggcc tgtacaatga actgcagaaa gataagatgg cggaggccta cagtgagatt     1260

gggatgaaag gcgagcgccg gaggggcaag gggcacgatg gcctttacca gggtctcagt     1320

acagccacca aggacaccta cgacgccctt cacatgcagg ccctgcctcc tcgcagagcc     1380

gagggcaggg gaagtcttct aacatgcggg gacgtggagg aaaatcccgg gcccatggcc     1440

gtgcccactc aggtcctggg gttgttgcta ctgtggctta cagatgccag atgtgacatc     1500

cagatgacac agtctccatc ttccctgtct gcatctgtcg gagatcgcgt caccatcacc     1560

tgtcgagcaa gtgaggacat ttattttaat ttagtgtggt atcagcagaa accaggaaag     1620

gcccctaagc tcctgatcta tgatacaaat cgcttggcag atggggtccc atcacggttc     1680

agtggctctg gatctggcac acagtatact ctaaccataa gtagcctgca acccgaagat     1740

ttcgcaacct attattgtca acactataag aattatccgc tcacgttcgg tcaggggacc     1800

aagctggaaa tcaaaagatc tggtggcgga gggtcaggag gcggaggcag cggaggcggt     1860

ggctcgggag gcggaggctc gagatctgag gtgcagttgg tggagtctgg gggcggcttg     1920

gtgcagcctg gagggtccct gaggctctcc tgtgcagcct caggattcac tctcagtaat     1980

tatggcatgc actggatcag gcaggctcca gggaagggtc tggagtgggt ctcgtctatt     2040

agtcttaatg gtggtagcac ttactatcga gactccgtga agggccgatt cactatctcc     2100

agggacaatg caaaaagcac cctctacctt caaatgaata gtctgagggc cgaggacacg     2160

gccgtctatt actgtgcagc acaggacgct tatacgggag gttactttga ttactggggc     2220

caaggaacgc tggtcacagt ctcgtctatg gatcccgcca ccacaaccaa gcccgtgctg     2280

cggaccccaa gccctgtgca ccctaccggc accagccagc ctcagagacc cgaggactgc     2340

cggcctcggg gcagcgtgaa gggcaccggc ctggacttcg cctgcgacat ctactgggca     2400

cctctggccg gaatatgcgt ggcactgctg ctgagcctca tcatcaccct gatctgttat     2460

caccgaagcc gcaagcgggt gtgtaaaagc gccattcagg ccgcttggcc ttctggcaca     2520

gagtgtatcg ccaagtacaa cttccacggc accgccgagc aggacctgcc tttctgtaaa     2580

ggcgacgtgc tgaccatcgt ggccgtgaca aaggacccca actggtacaa ggccaagaac     2640

aaagtgggca gagagggcat catccccgcc aactatgtgc agaagagaga gggcgttaag     2700

gccggcacca agctgtctct gatgccctgg tttcacggca agatcaccag agagcaggcc     2760

gagagactgc tgtaccctcc tgaaaccggc ctgttcctcg tgcgcgagag cacaaattac     2820

cccggcgact acaccctgtg tgtgtcctgt gatggcaagg tggaacacta ccggatcatg     2880

taccacgcca gcaagctgag catcgacgag gaagtgtact tcgagaacct gatgcagctg     2940

gtcgagcact acacctccga tgccgatggc ctgtgcacca gactgatcaa gcccaaagtg     3000

atggaaggca ccgtggccgc tcaggacgag ttttacagat ccggctgggc tctgaacatg     3060

aaggaactga agctgctgca gaccatcggc aagggcgagt ttggagatgt gatgctgggc     3120

gactaccggg gcaacaaggt ggcagtgaag tgcatcaaga acgacgctac agcccaggct     3180

tttctggccg aagccagcgt gatgacccag ctgagacaca gcaatctggt gcagctgctg     3240

ggcgtgatcg tggaagaaaa aggcggcctg tatatcgtga ccgagtacat ggccaagggc     3300

agcctggtgg actacctgag aagtagaggc agaagcgtgc tcggaggcga ctgcctgctg     3360

aagtttagcc tggatgtgtg cgaggctatg gaatacctgg aaggcaacaa cttcgtgcac     3420

cgcgatctgg ccgccagaaa tgtgctggtg tccgaggaca acgtggccaa ggtgtccgat     3480

ttcggcctga ccaaagaggc cagcagcacc caggatacag gcaagctgcc cgtgaaatgg     3540

acagcccctg aggctctgag agagaagaag ttcagcacca agagcgacgt gtggtccttc     3600

ggcatcctgc tgtgggaaat ctacagcttc ggcagagtgc cctatcctag aatccctctg     3660

aaggacgtgg tgcccagagt ggaaaagggc tacaagatgg atgcccctga cggctgtcct     3720

cctgccgtgt acgaagtgat gaagaactgc tggcacctgg acgccgctat gaggccatct     3780

ttcctgcagc tgagagagca gctggaacac atcaagaccc acgagctgca cctg           3834


<210>  20
<211>  967
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  sequence of CAR construct comprising tyrosine kinase domain of 
       CSK (tkCSK)

<400>  20

Met Ser Leu Pro Val Thr Ala Leu Leu Leu Pro Leu Ala Leu Leu Leu 
1               5                   10                  15      


His Ala Ala Arg Pro Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu 
            20                  25                  30          


Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln 
        35                  40                  45              


Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr 
    50                  55                  60                  


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


Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile 
                85                  90                  95      


Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly 
            100                 105                 110         


Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr 
        115                 120                 125             


Lys Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 
    130                 135                 140                 


Ser Gly Gly Gly Gly Ser Glu Val Lys Leu Gln Glu Ser Gly Pro Gly 
145                 150                 155                 160 


Leu Val Ala Pro Ser Gln Ser Leu Ser Val Thr Cys Thr Val Ser Gly 
                165                 170                 175     


Val Ser Leu Pro Asp Tyr Gly Val Ser Trp Ile Arg Gln Pro Pro Arg 
            180                 185                 190         


Lys Gly Leu Glu Trp Leu Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr 
        195                 200                 205             


Tyr Asn Ser Ala Leu Lys Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser 
    210                 215                 220                 


Lys Ser Gln Val Phe Leu Lys Met Asn Ser Leu Gln Thr Asp Asp Thr 
225                 230                 235                 240 


Ala Ile Tyr Tyr Cys Ala Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala 
                245                 250                 255     


Met Asp Tyr Trp Gly Gln Gly Thr Ser Val Thr Val Ser Ser Asp Pro 
            260                 265                 270         


Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala 
        275                 280                 285             


Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly 
    290                 295                 300                 


Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Phe Trp 
305                 310                 315                 320 


Val Leu Val Val Val Gly Gly Val Leu Ala Cys Tyr Ser Leu Leu Val 
                325                 330                 335     


Thr Val Ala Phe Ile Ile Phe Trp Val Arg Arg Val Lys Phe Ser Arg 
            340                 345                 350         


Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn 
        355                 360                 365             


Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg 
    370                 375                 380                 


Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro 
385                 390                 395                 400 


Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala 
                405                 410                 415     


Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His 
            420                 425                 430         


Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp 
        435                 440                 445             


Ala Leu His Met Gln Ala Leu Pro Pro Arg Arg Ala Glu Gly Arg Gly 
    450                 455                 460                 


Ser Leu Leu Thr Cys Gly Asp Val Glu Glu Asn Pro Gly Pro Met Ala 
465                 470                 475                 480 


Val Pro Thr Gln Val Leu Gly Leu Leu Leu Leu Trp Leu Thr Asp Ala 
                485                 490                 495     


Arg Cys Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser 
            500                 505                 510         


Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Glu Asp Ile Tyr 
        515                 520                 525             


Phe Asn Leu Val Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu 
    530                 535                 540                 


Leu Ile Tyr Asp Thr Asn Arg Leu Ala Asp Gly Val Pro Ser Arg Phe 
545                 550                 555                 560 


Ser Gly Ser Gly Ser Gly Thr Gln Tyr Thr Leu Thr Ile Ser Ser Leu 
                565                 570                 575     


Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln His Tyr Lys Asn Tyr 
            580                 585                 590         


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


Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 
    610                 615                 620                 


Gly Gly Ser Arg Ser Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu 
625                 630                 635                 640 


Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe 
                645                 650                 655     


Thr Leu Ser Asn Tyr Gly Met His Trp Ile Arg Gln Ala Pro Gly Lys 
            660                 665                 670         


Gly Leu Glu Trp Val Ser Ser Ile Ser Leu Asn Gly Gly Ser Thr Tyr 
        675                 680                 685             


Tyr Arg Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala 
    690                 695                 700                 


Lys Ser Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr 
705                 710                 715                 720 


Ala Val Tyr Tyr Cys Ala Ala Gln Asp Ala Tyr Thr Gly Gly Tyr Phe 
                725                 730                 735     


Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Met Asp Pro 
            740                 745                 750         


Ala Thr Thr Thr Lys Pro Val Leu Arg Thr Pro Ser Pro Val His Pro 
        755                 760                 765             


Thr Gly Thr Ser Gln Pro Gln Arg Pro Glu Asp Cys Arg Pro Arg Gly 
    770                 775                 780                 


Ser Val Lys Gly Thr Gly Leu Asp Phe Ala Cys Asp Ile Tyr Trp Ala 
785                 790                 795                 800 


Pro Leu Ala Gly Ile Cys Val Ala Leu Leu Leu Ser Leu Ile Ile Thr 
                805                 810                 815     


Leu Ile Cys Tyr His Arg Ser Arg Lys Arg Val Cys Lys Leu Lys Leu 
            820                 825                 830         


Leu Gln Thr Ile Gly Lys Gly Glu Phe Gly Asp Val Met Leu Gly Asp 
        835                 840                 845             


Tyr Arg Gly Asn Lys Val Ala Val Lys Cys Ile Lys Asn Asp Ala Thr 
    850                 855                 860                 


Ala Gln Ala Phe Leu Ala Glu Ala Ser Val Met Thr Gln Leu Arg His 
865                 870                 875                 880 


Ser Asn Leu Val Gln Leu Leu Gly Val Ile Val Glu Glu Lys Gly Gly 
                885                 890                 895     


Leu Tyr Ile Val Thr Glu Tyr Met Ala Lys Gly Ser Leu Val Asp Tyr 
            900                 905                 910         


Leu Arg Ser Arg Gly Arg Ser Val Leu Gly Gly Asp Cys Leu Leu Lys 
        915                 920                 925             


Phe Ser Leu Asp Val Cys Glu Ala Met Glu Tyr Leu Glu Gly Asn Asn 
    930                 935                 940                 


Phe Val His Arg Asp Leu Ala Ala Arg Asn Val Leu Val Ser Glu Asp 
945                 950                 955                 960 


Asn Val Ala Lys Val Ser Asp 
                965         


<210>  21
<211>  4
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  ITAM motif


<220>
<221>  misc_feature
<222>  (2)..(3)
<223>  Xaa can be any naturally occurring amino acid

<220>
<221>  MISC_FEATURE
<222>  (4)..(4)
<223>  Xaa may be Leu or Ile

<400>  21

Tyr Xaa Xaa Xaa 
1               


