               SEQUENCE LISTING

<110> Ludwig-Maximilians-Universität München

<120> CD28-targeting chimeric antigen receptor (CAR) T cells, methods of generation and uses thereof

<130> AD2709 PCT

<150> EP 20 209 327.4
<151> 2020-11-23

<160> 58

<170> BiSSAP 1.3.6

<210> 1
<211> 5
<212> PRT
<213> Artificial Sequence


<220> 
<223> possible sequence unit of hinge or linker (GGGGS)n

<400> 1
Gly Gly Gly Gly Ser 
1               5   

<210> 2
<211> 18
<212> PRT
<213> Artificial Sequence


<220> 
<223> Whitlow linker

<400> 2
Gly Ser Thr Ser Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr 
1               5                   10                  15      
Lys Gly 
        

<210> 3
<211> 21
<212> PRT
<213> Homo sapiens


<220> 
<223> human CD8 alpha signal peptide

<400> 3
Met Ala 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> 4
<211> 22
<212> PRT
<213> Homo sapiens


<220> 
<223> signal peptide of human Colony Stimulating Factor 2 Receptor
      Subunit Alpha (CSF2RA)

<400> 4
Met Leu Leu Leu Val Thr Ser Leu Leu Leu Cys Glu Leu Pro His Pro 
1               5                   10                  15      
Ala Phe Leu Leu Ile Pro 
            20          

<210> 5
<211> 55
<212> PRT
<213> Homo sapiens


<220> 
<223> exemplary “long” hinge (human CD8 alpha extracellular portion)

<400> 5
Phe Val Pro Val Phe Leu Pro Ala Lys Pro Thr Thr Thr Pro Ala Pro 
1               5                   10                  15      
Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu 
            20                  25                  30          
Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly Gly Ala Val His Thr Arg 
        35                  40                  45              
Gly Leu Asp Phe Ala Cys Asp 
    50                  55  

<210> 6
<211> 15
<212> PRT
<213> Artificial Sequence


<220> 
<223> exemplary “short” hinge: (GGGGS)n, n = 3

<400> 6
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 
1               5                   10                  15  

<210> 7
<211> 10
<212> PRT
<213> Artificial Sequence


<220> 
<223> myc-tag

<400> 7
Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu 
1               5                   10  

<210> 8
<211> 5
<212> PRT
<213> Artificial Sequence


<220> 
<223> monoclonal antibody TGN1412 - VH CDR1

<400> 8
Ser Tyr Tyr Ile His 
1               5   

<210> 9
<211> 17
<212> PRT
<213> Artificial Sequence


<220> 
<223> monoclonal antibody TGN1412 - VH CDR2

<400> 9
Cys Ile Tyr Pro Gly Asn Val Asn Thr Asn Tyr Asn Glu Lys Phe Lys 
1               5                   10                  15      
Asp 
    

<210> 10
<211> 11
<212> PRT
<213> Artificial Sequence


<220> 
<223> monoclonal antibody TGN1412 - VH CDR3

<400> 10
Ser His Tyr Gly Leu Asp Trp Asn Phe Asp Val 
1               5                   10      

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


<220> 
<223> monoclonal antibody TGN1412 - VL CDR1

<400> 11
His Ala Ser Gln Asn Ile Tyr Val Trp Leu Asn 
1               5                   10      

<210> 12
<211> 7
<212> PRT
<213> Artificial Sequence


<220> 
<223> monoclonal antibody TGN1412 - VL CDR2

<400> 12
Lys Ala Ser Asn Leu His Thr 
1               5           

<210> 13
<211> 9
<212> PRT
<213> Artificial Sequence


<220> 
<223> monoclonal antibody TGN1412 - VL CDR3

<400> 13
Gln Gln Gly Gln Thr Tyr Pro Tyr Thr 
1               5                   

<210> 14
<211> 5
<212> PRT
<213> Artificial Sequence


<220> 
<223> monoclonal antibody CD28.3 - VH CDR1

<400> 14
Glu Tyr Ile Ile His 
1               5   

<210> 15
<211> 17
<212> PRT
<213> Artificial Sequence


<220> 
<223> monoclonal antibody CD28.3 - VH CDR2

<400> 15
Trp Phe Tyr Pro Gly Ser Asn Asp Ile Gln Tyr Asn Ala Lys Phe Lys 
1               5                   10                  15      
Gly 
    

<210> 16
<211> 12
<212> PRT
<213> Artificial Sequence


<220> 
<223> monoclonal antibody CD28.3 - VH CDR3

<400> 16
Arg Asp Asp Phe Ser Gly Tyr Asp Ala Leu Pro Tyr 
1               5                   10          

<210> 17
<211> 11
<212> PRT
<213> Artificial Sequence


<220> 
<223> monoclonal antibody CD28.3 - VL CDR1

<400> 17
Arg Thr Asn Glu Asn Ile Tyr Ser Asn Leu Ala 
1               5                   10      

<210> 18
<211> 7
<212> PRT
<213> Artificial Sequence


<220> 
<223> monoclonal antibody CD28.3 - VL CDR2

<400> 18
Ala Ala Thr His Leu Val Glu 
1               5           

<210> 19
<211> 9
<212> PRT
<213> Artificial Sequence


<220> 
<223> monoclonal antibody CD28.3 - VL CDR3

<400> 19
Gln His Phe Trp Gly Thr Pro Cys Thr 
1               5                   

<210> 20
<211> 120
<212> PRT
<213> Artificial Sequence


<220> 
<223> monoclonal antibody TGN1412 VH

<400> 20
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 
1               5                   10                  15      
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 
            20                  25                  30          
Tyr Ile His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 
        35                  40                  45              
Gly Cys Ile Tyr Pro Gly Asn Val Asn Thr Asn Tyr Asn Glu Lys Phe 
    50                  55                  60                  
Lys Asp Arg Ala Thr Leu Thr Val Asp Thr Ser Ile Ser Thr Ala Tyr 
65                  70                  75                  80  
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Phe Cys 
                85                  90                  95      
Thr Arg Ser His Tyr Gly Leu Asp Trp Asn Phe Asp Val Trp Gly Gln 
            100                 105                 110         
Gly Thr Thr Val Thr Val Ser Ser 
        115                 120 

<210> 21
<211> 110
<212> PRT
<213> Artificial Sequence


<220> 
<223> monoclonal antibody TGN1412 VL

<400> 21
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 
1               5                   10                  15      
Asp Arg Val Thr Ile Thr Cys His Ala Ser Gln Asn Ile Tyr Val Trp 
            20                  25                  30          
Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 
        35                  40                  45              
Tyr Lys Ala Ser Asn Leu His Thr Gly Val Pro Ser Arg Phe Ser Gly 
    50                  55                  60                  
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 
65                  70                  75                  80  
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Gly Gln Thr Tyr Pro Tyr 
                85                  90                  95      
Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val 
            100                 105                 110 

<210> 22
<211> 120
<212> PRT
<213> Artificial Sequence


<220> 
<223> monoclonal antibody CD28.3 - VH

<400> 22
Val Lys Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala Ser 
1               5                   10                  15      
Val Arg Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Tyr Ile 
            20                  25                  30          
Ile His Trp Ile Lys Leu Arg Ser Gly Gln Gly Leu Glu Trp Ile Gly 
        35                  40                  45              
Trp Phe Tyr Pro Gly Ser Asn Asp Ile Gln Tyr Asn Ala Lys Phe Lys 
    50                  55                  60                  
Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Val Tyr Met 
65                  70                  75                  80  
Glu Leu Thr Gly Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys Ala 
                85                  90                  95      
Arg Arg Asp Asp Phe Ser Gly Tyr Asp Ala Leu Pro Tyr Trp Gly Gln 
            100                 105                 110         
Gly Thr Met Val Thr Val Ser Ser 
        115                 120 

<210> 23
<211> 108
<212> PRT
<213> Artificial Sequence


<220> 
<223> monoclonal antibody CD28.3 - VL

<400> 23
Asp Ile Gln Met Thr Gln Ser Pro Ala Ser Leu Ser Val Ser Val Gly 
1               5                   10                  15      
Glu Thr Val Thr Ile Thr Cys Arg Thr Asn Glu Asn Ile Tyr Ser Asn 
            20                  25                  30          
Leu Ala Trp Tyr Gln Gln Lys Gln Gly Lys Ser Pro Gln Leu Leu Ile 
        35                  40                  45              
Tyr Ala Ala Thr His Leu Val Glu Gly Val Pro Ser Arg Phe Ser Gly 
    50                  55                  60                  
Ser Gly Ser Gly Thr Gln Tyr Ser Leu Lys Ile Thr Ser Leu Gln Ser 
65                  70                  75                  80  
Glu Asp Phe Gly Asn Tyr Tyr Cys Gln His Phe Trp Gly Thr Pro Cys 
                85                  90                  95      
Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg 
            100                 105             

<210> 24
<211> 248
<212> PRT
<213> Artificial Sequence


<220> 
<223> scFv of construct CD28_CAR_2 (without signal peptide)

<400> 24
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 
1               5                   10                  15      
Asp Arg Val Thr Ile Thr Cys His Ala Ser Gln Asn Ile Tyr Val Trp 
            20                  25                  30          
Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 
        35                  40                  45              
Tyr Lys Ala Ser Asn Leu His Thr Gly Val Pro Ser Arg Phe Ser Gly 
    50                  55                  60                  
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 
65                  70                  75                  80  
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Gly Gln Thr Tyr Pro Tyr 
                85                  90                  95      
Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Gly Ser 
            100                 105                 110         
Thr Ser Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys Gly 
        115                 120                 125             
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 
    130                 135                 140                 
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 
145                 150                 155                 160 
Tyr Ile His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 
                165                 170                 175     
Gly Cys Ile Tyr Pro Gly Asn Val Asn Thr Asn Tyr Asn Glu Lys Phe 
            180                 185                 190         
Lys Asp Arg Ala Thr Leu Thr Val Asp Thr Ser Ile Ser Thr Ala Tyr 
        195                 200                 205             
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Phe Cys 
    210                 215                 220                 
Thr Arg Ser His Tyr Gly Leu Asp Trp Asn Phe Asp Val Trp Gly Gln 
225                 230                 235                 240 
Gly Thr Thr Val Thr Val Ser Ser 
                245             

<210> 25
<211> 248
<212> PRT
<213> Artificial Sequence


<220> 
<223> scFv of construct CD28_CAR_12 (without signal peptide)

<400> 25
Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 
1               5                   10                  15      
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 
            20                  25                  30          
Tyr Ile His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 
        35                  40                  45              
Gly Cys Ile Tyr Pro Gly Asn Val Asn Thr Asn Tyr Asn Glu Lys Phe 
    50                  55                  60                  
Lys Asp Arg Ala Thr Leu Thr Val Asp Thr Ser Ile Ser Thr Ala Tyr 
65                  70                  75                  80  
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Phe Cys 
                85                  90                  95      
Thr Arg Ser His Tyr Gly Leu Asp Trp Asn Phe Asp Val Trp Gly Gln 
            100                 105                 110         
Gly Thr Thr Val Thr Val Ser Ser Gly Ser Thr Ser Gly Ser Gly Lys 
        115                 120                 125             
Pro Gly Ser Gly Glu Gly Ser Thr Lys Gly Asp Ile Gln Met Thr Gln 
    130                 135                 140                 
Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr 
145                 150                 155                 160 
Cys His Ala Ser Gln Asn Ile Tyr Val Trp Leu Asn Trp Tyr Gln Gln 
                165                 170                 175     
Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr Lys Ala Ser Asn Leu 
            180                 185                 190         
His Thr Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp 
        195                 200                 205             
Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr 
    210                 215                 220                 
Tyr Cys Gln Gln Gly Gln Thr Tyr Pro Tyr Thr Phe Gly Gly Gly Thr 
225                 230                 235                 240 
Lys Val Glu Ile Lys Arg Thr Val 
                245             

<210> 26
<211> 246
<212> PRT
<213> Artificial Sequence


<220> 
<223> scFv of construct CD28_CAR_14 (without signal peptide)

<400> 26
Val Lys Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala Ser 
1               5                   10                  15      
Val Arg Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Glu Tyr Ile 
            20                  25                  30          
Ile His Trp Ile Lys Leu Arg Ser Gly Gln Gly Leu Glu Trp Ile Gly 
        35                  40                  45              
Trp Phe Tyr Pro Gly Ser Asn Asp Ile Gln Tyr Asn Ala Lys Phe Lys 
    50                  55                  60                  
Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Val Tyr Met 
65                  70                  75                  80  
Glu Leu Thr Gly Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys Ala 
                85                  90                  95      
Arg Arg Asp Asp Phe Ser Gly Tyr Asp Ala Leu Pro Tyr Trp Gly Gln 
            100                 105                 110         
Gly Thr Met Val Thr Val Ser Ser Gly Ser Thr Ser Gly Ser Gly Lys 
        115                 120                 125             
Pro Gly Ser Gly Glu Gly Ser Thr Lys Gly Asp Ile Gln Met Thr Gln 
    130                 135                 140                 
Ser Pro Ala Ser Leu Ser Val Ser Val Gly Glu Thr Val Thr Ile Thr 
145                 150                 155                 160 
Cys Arg Thr Asn Glu Asn Ile Tyr Ser Asn Leu Ala Trp Tyr Gln Gln 
                165                 170                 175     
Lys Gln Gly Lys Ser Pro Gln Leu Leu Ile Tyr Ala Ala Thr His Leu 
            180                 185                 190         
Val Glu Gly Val Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Gln 
        195                 200                 205             
Tyr Ser Leu Lys Ile Thr Ser Leu Gln Ser Glu Asp Phe Gly Asn Tyr 
    210                 215                 220                 
Tyr Cys Gln His Phe Trp Gly Thr Pro Cys Thr Phe Gly Gly Gly Thr 
225                 230                 235                 240 
Lys Leu Glu Ile Lys Arg 
                245     

<210> 27
<211> 41
<212> PRT
<213> Homo sapiens


<220> 
<223> human CD28 signaling domain

<400> 27
Arg Ser Lys Arg Ser Arg Leu Leu His Ser Asp Tyr Met Asn Met Thr 
1               5                   10                  15      
Pro Arg Arg Pro Gly Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala Pro 
            20                  25                  30          
Pro Arg Asp Phe Ala Ala Tyr Arg Ser 
        35                  40      

<210> 28
<211> 41
<212> PRT
<213> Mus musculus


<220> 
<223> murine CD28 signaling domain

<400> 28
Asn Ser Arg Arg Asn Arg Leu Leu Gln Ser Asp Tyr Met Asn Met Thr 
1               5                   10                  15      
Pro Arg Arg Pro Gly Leu Thr Arg Lys Pro Tyr Gln Pro Tyr Ala Pro 
            20                  25                  30          
Ala Arg Asp Phe Ala Ala Tyr Arg Pro 
        35                  40      

<210> 29
<211> 41
<212> PRT
<213> Rattus norvegicus


<220> 
<223> rat CD28 signaling domain

<400> 29
Asn Ser Arg Arg Asn Arg Leu Leu Gln Ser Asp Tyr Met Asn Met Thr 
1               5                   10                  15      
Pro Arg Arg Leu Gly Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala Pro 
            20                  25                  30          
Ala Arg Asp Phe Ala Ala Tyr Arg Pro 
        35                  40      

<210> 30
<211> 112
<212> PRT
<213> Homo sapiens


<220> 
<223> human CD3ζ signaling domain

<400> 30
Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly 
1               5                   10                  15      
Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr 
            20                  25                  30          
Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys 
        35                  40                  45              
Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys 
    50                  55                  60                  
Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg 
65                  70                  75                  80  
Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala 
                85                  90                  95      
Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg 
            100                 105                 110         



<210> 31
<211> 143
<212> PRT
<213> Mus musculus


<220> 
<223> murine CD3ζ signaling domain

<400> 31
Gln Ser Phe Gly Leu Leu Asp Pro Lys Leu Cys Tyr Leu Leu Asp Gly 
1               5                   10                  15      
Ile Leu Phe Ile Tyr Gly Val Ile Ile Thr Ala Leu Tyr Leu Arg Ala 
            20                  25                  30          
Lys Phe Ser Arg Ser Ala Glu Thr Ala Ala Asn Leu Gln Asp Pro Asn 
        35                  40                  45              
Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val 
    50                  55                  60                  
Leu Glu Lys Lys Arg Ala Arg Asp Pro Glu Met Gly Gly Lys Gln Gln 
65                  70                  75                  80  
Arg Arg Arg Asn Pro Gln Glu Gly Val Tyr Asn Ala Leu Gln Lys Asp 
                85                  90                  95      
Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Thr Lys Gly Glu Arg Arg 
            100                 105                 110         
Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr 
        115                 120                 125             
Lys Asp Thr Tyr Asp Ala Leu His Met Gln Thr Leu Ala Pro Arg 
    130                 135                 140             

<210> 32
<211> 143
<212> PRT
<213> Rattus norvegicus


<220> 
<223> rat CD3ζ signaling domain

<400> 32
Gln Ser Phe Gly Leu Leu Asp Pro Lys Leu Cys Tyr Met Leu Asp Gly 
1               5                   10                  15      
Ile Leu Phe Ile Tyr Gly Val Ile Val Thr Ala Leu Tyr Leu Arg Ala 
            20                  25                  30          
Lys Phe Ser Arg Ser Ala Asp Ala Ala Ala Tyr Leu Gln Asp Pro Asn 
        35                  40                  45              
Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val 
    50                  55                  60                  
Leu Asp Lys Lys Arg Pro Arg Asp Pro Glu Met Gly Gly Lys Gln Gln 
65                  70                  75                  80  
Arg Arg Arg Asn Pro Gln Glu Gly Val Tyr Asn Ala Leu Gln Lys Asp 
                85                  90                  95      
Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg 
            100                 105                 110         
Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr 
        115                 120                 125             
Lys Asp Thr Tyr Asp Ala Leu His Met Gln Thr Leu Pro Pro Arg 
    130                 135                 140             

<210> 33
<211> 21
<212> PRT
<213> Homo sapiens


<220> 
<223> human CD8 alpha transmembrane domain

<400> 33
Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu 
1               5                   10                  15      
Ser Leu Val Ile Thr 
            20      

<210> 34
<211> 21
<212> PRT
<213> Mus musculus


<220> 
<223> murine CD8 alpha transmembrane domain

<400> 34
Ile Trp Ala Pro Leu Ala Gly Ile Cys Val Ala Leu Leu Leu Ser Leu 
1               5                   10                  15      
Ile Ile Thr Leu Ile 
            20      

<210> 35
<211> 21
<212> PRT
<213> Rattus norvegicus


<220> 
<223> rat CD8 alpha transmembrane domain

<400> 35
Ile Trp Ala Pro Leu Ala Gly Ile Cys Ala Val Leu Leu Leu Ser Leu 
1               5                   10                  15      
Val Ile Thr Leu Ile 
            20      

<210> 36
<211> 7
<212> PRT
<213> Homo sapiens


<220> 
<223> human CD8 alpha intracellular peptide

<400> 36
Leu Tyr Cys Asn His Arg Asn 
1               5           

<210> 37
<211> 7
<212> PRT
<213> Mus musculus


<220> 
<223> murine CD8 alpha intracellular peptide

<400> 37
Cys Tyr His Arg Ser Arg Lys 
1               5           

<210> 38
<211> 7
<212> PRT
<213> Rattus norvegicus


<220> 
<223> rat CD8 alpha intracellular peptide

<400> 38
Cys Cys His Arg Asn Arg Arg 
1               5           

<210> 39
<211> 181
<212> PRT
<213> Artificial Sequence


<220> 
<223> hCD8 alpha TM + intracell. peptide, hCD28 signaling domain, hCD3ζ
      signaling domain

<400> 39
Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu 
1               5                   10                  15      
Ser Leu Val Ile Thr Leu Tyr Cys Asn His Arg Asn Arg Ser Lys Arg 
            20                  25                  30          
Ser Arg Leu Leu His Ser Asp Tyr Met Asn Met Thr Pro Arg Arg Pro 
        35                  40                  45              
Gly Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala Pro Pro Arg Asp Phe 
    50                  55                  60                  
Ala Ala Tyr Arg Ser Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro 
65                  70                  75                  80  
Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly 
                85                  90                  95      
Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro 
            100                 105                 110         
Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr 
        115                 120                 125             
Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly 
    130                 135                 140                 
Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln 
145                 150                 155                 160 
Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln 
                165                 170                 175     
Ala Leu Pro Pro Arg 
            180     

<210> 40
<211> 476
<212> PRT
<213> Artificial Sequence


<220> 
<223> CD28_CAR_1 (including signal peptide)

<400> 40
Met Leu Leu Leu Val Thr Ser Leu Leu Leu Cys Glu Leu Pro His Pro 
1               5                   10                  15      
Ala Phe Leu Leu Ile Pro Asp Ile Gln Met Thr Gln Ser Pro Ser Ser 
            20                  25                  30          
Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys His Ala Ser 
        35                  40                  45              
Gln Asn Ile Tyr Val Trp Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys 
    50                  55                  60                  
Ala Pro Lys Leu Leu Ile Tyr Lys Ala Ser Asn Leu His Thr Gly Val 
65                  70                  75                  80  
Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 
                85                  90                  95      
Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln 
            100                 105                 110         
Gly Gln Thr Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Val Glu Ile 
        115                 120                 125             
Lys Arg Thr Val Gly Ser Thr Ser Gly Ser Gly Lys Pro Gly Ser Gly 
    130                 135                 140                 
Glu Gly Ser Thr Lys Gly Gln Val Gln Leu Val Gln Ser Gly Ala Glu 
145                 150                 155                 160 
Val Lys Lys Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly 
                165                 170                 175     
Tyr Thr Phe Thr Ser Tyr Tyr Ile His Trp Val Arg Gln Ala Pro Gly 
            180                 185                 190         
Gln Gly Leu Glu Trp Ile Gly Cys Ile Tyr Pro Gly Asn Val Asn Thr 
        195                 200                 205             
Asn Tyr Asn Glu Lys Phe Lys Asp Arg Ala Thr Leu Thr Val Asp Thr 
    210                 215                 220                 
Ser Ile Ser Thr Ala Tyr Met Glu Leu Ser Arg Leu Arg Ser Asp Asp 
225                 230                 235                 240 
Thr Ala Val Tyr Phe Cys Thr Arg Ser His Tyr Gly Leu Asp Trp Asn 
                245                 250                 255     
Phe Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Glu Gln 
            260                 265                 270         
Lys Leu Ile Ser Glu Glu Asp Leu Gly Gly Gly Gly Ser Gly Gly Gly 
        275                 280                 285             
Gly Ser Gly Gly Gly Gly Ser Ile Tyr Ile Trp Ala Pro Leu Ala Gly 
    290                 295                 300                 
Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Asn 
305                 310                 315                 320 
His Arg Asn Arg Ser Lys Arg Ser Arg Leu Leu His Ser Asp Tyr Met 
                325                 330                 335     
Asn Met Thr Pro Arg Arg Pro Gly Pro Thr Arg Lys His Tyr Gln Pro 
            340                 345                 350         
Tyr Ala Pro Pro Arg Asp Phe Ala Ala Tyr Arg Ser Arg Val Lys Phe 
        355                 360                 365             
Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu 
    370                 375                 380                 
Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp 
385                 390                 395                 400 
Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys 
                405                 410                 415     
Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala 
            420                 425                 430         
Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys 
        435                 440                 445             
Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr 
    450                 455                 460                 
Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg 
465                 470                 475     

<210> 41
<211> 516
<212> PRT
<213> Artificial Sequence


<220> 
<223> CD28_CAR_2 (including signal peptide)

<400> 41
Met Leu Leu Leu Val Thr Ser Leu Leu Leu Cys Glu Leu Pro His Pro 
1               5                   10                  15      
Ala Phe Leu Leu Ile Pro Asp Ile Gln Met Thr Gln Ser Pro Ser Ser 
            20                  25                  30          
Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys His Ala Ser 
        35                  40                  45              
Gln Asn Ile Tyr Val Trp Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys 
    50                  55                  60                  
Ala Pro Lys Leu Leu Ile Tyr Lys Ala Ser Asn Leu His Thr Gly Val 
65                  70                  75                  80  
Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 
                85                  90                  95      
Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln 
            100                 105                 110         
Gly Gln Thr Tyr Pro Tyr Thr Phe Gly Gly Gly Thr Lys Val Glu Ile 
        115                 120                 125             
Lys Arg Thr Val Gly Ser Thr Ser Gly Ser Gly Lys Pro Gly Ser Gly 
    130                 135                 140                 
Glu Gly Ser Thr Lys Gly Gln Val Gln Leu Val Gln Ser Gly Ala Glu 
145                 150                 155                 160 
Val Lys Lys Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly 
                165                 170                 175     
Tyr Thr Phe Thr Ser Tyr Tyr Ile His Trp Val Arg Gln Ala Pro Gly 
            180                 185                 190         
Gln Gly Leu Glu Trp Ile Gly Cys Ile Tyr Pro Gly Asn Val Asn Thr 
        195                 200                 205             
Asn Tyr Asn Glu Lys Phe Lys Asp Arg Ala Thr Leu Thr Val Asp Thr 
    210                 215                 220                 
Ser Ile Ser Thr Ala Tyr Met Glu Leu Ser Arg Leu Arg Ser Asp Asp 
225                 230                 235                 240 
Thr Ala Val Tyr Phe Cys Thr Arg Ser His Tyr Gly Leu Asp Trp Asn 
                245                 250                 255     
Phe Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Glu Gln 
            260                 265                 270         
Lys Leu Ile Ser Glu Glu Asp Leu Phe Val Pro Val Phe Leu Pro Ala 
        275                 280                 285             
Lys Pro Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr 
    290                 295                 300                 
Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala 
305                 310                 315                 320 
Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile 
                325                 330                 335     
Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser 
            340                 345                 350         
Leu Val Ile Thr Leu Tyr Cys Asn His Arg Asn Arg Ser Lys Arg Ser 
        355                 360                 365             
Arg Leu Leu His Ser Asp Tyr Met Asn Met Thr Pro Arg Arg Pro Gly 
    370                 375                 380                 
Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala Pro Pro Arg Asp Phe Ala 
385                 390                 395                 400 
Ala Tyr Arg Ser Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala 
                405                 410                 415     
Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg 
            420                 425                 430         
Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu 
        435                 440                 445             
Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn 
    450                 455                 460                 
Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met 
465                 470                 475                 480 
Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly 
                485                 490                 495     
Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala 
            500                 505                 510         
Leu Pro Pro Arg 
        515     

<210> 42
<211> 476
<212> PRT
<213> Artificial Sequence


<220> 
<223> CD28_CAR_11 (including signal peptide)

<400> 42
Met Leu Leu Leu Val Thr Ser Leu Leu Leu Cys Glu Leu Pro His Pro 
1               5                   10                  15      
Ala Phe Leu Leu Ile Pro Gln Val Gln Leu Val Gln Ser Gly Ala Glu 
            20                  25                  30          
Val Lys Lys Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly 
        35                  40                  45              
Tyr Thr Phe Thr Ser Tyr Tyr Ile His Trp Val Arg Gln Ala Pro Gly 
    50                  55                  60                  
Gln Gly Leu Glu Trp Ile Gly Cys Ile Tyr Pro Gly Asn Val Asn Thr 
65                  70                  75                  80  
Asn Tyr Asn Glu Lys Phe Lys Asp Arg Ala Thr Leu Thr Val Asp Thr 
                85                  90                  95      
Ser Ile Ser Thr Ala Tyr Met Glu Leu Ser Arg Leu Arg Ser Asp Asp 
            100                 105                 110         
Thr Ala Val Tyr Phe Cys Thr Arg Ser His Tyr Gly Leu Asp Trp Asn 
        115                 120                 125             
Phe Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Ser 
    130                 135                 140                 
Thr Ser Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys Gly 
145                 150                 155                 160 
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 
                165                 170                 175     
Asp Arg Val Thr Ile Thr Cys His Ala Ser Gln Asn Ile Tyr Val Trp 
            180                 185                 190         
Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 
        195                 200                 205             
Tyr Lys Ala Ser Asn Leu His Thr Gly Val Pro Ser Arg Phe Ser Gly 
    210                 215                 220                 
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 
225                 230                 235                 240 
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Gly Gln Thr Tyr Pro Tyr 
                245                 250                 255     
Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Glu Gln 
            260                 265                 270         
Lys Leu Ile Ser Glu Glu Asp Leu Gly Gly Gly Gly Ser Gly Gly Gly 
        275                 280                 285             
Gly Ser Gly Gly Gly Gly Ser Ile Tyr Ile Trp Ala Pro Leu Ala Gly 
    290                 295                 300                 
Thr Cys Gly Val Leu Leu Leu Ser Leu Val Ile Thr Leu Tyr Cys Asn 
305                 310                 315                 320 
His Arg Asn Arg Ser Lys Arg Ser Arg Leu Leu His Ser Asp Tyr Met 
                325                 330                 335     
Asn Met Thr Pro Arg Arg Pro Gly Pro Thr Arg Lys His Tyr Gln Pro 
            340                 345                 350         
Tyr Ala Pro Pro Arg Asp Phe Ala Ala Tyr Arg Ser Arg Val Lys Phe 
        355                 360                 365             
Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu 
    370                 375                 380                 
Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp 
385                 390                 395                 400 
Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys 
                405                 410                 415     
Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala 
            420                 425                 430         
Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys 
        435                 440                 445             
Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr 
    450                 455                 460                 
Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg 
465                 470                 475     

<210> 43
<211> 516
<212> PRT
<213> Artificial Sequence


<220> 
<223> CD28_CAR_12 (including signal peptide)

<400> 43
Met Leu Leu Leu Val Thr Ser Leu Leu Leu Cys Glu Leu Pro His Pro 
1               5                   10                  15      
Ala Phe Leu Leu Ile Pro Gln Val Gln Leu Val Gln Ser Gly Ala Glu 
            20                  25                  30          
Val Lys Lys Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly 
        35                  40                  45              
Tyr Thr Phe Thr Ser Tyr Tyr Ile His Trp Val Arg Gln Ala Pro Gly 
    50                  55                  60                  
Gln Gly Leu Glu Trp Ile Gly Cys Ile Tyr Pro Gly Asn Val Asn Thr 
65                  70                  75                  80  
Asn Tyr Asn Glu Lys Phe Lys Asp Arg Ala Thr Leu Thr Val Asp Thr 
                85                  90                  95      
Ser Ile Ser Thr Ala Tyr Met Glu Leu Ser Arg Leu Arg Ser Asp Asp 
            100                 105                 110         
Thr Ala Val Tyr Phe Cys Thr Arg Ser His Tyr Gly Leu Asp Trp Asn 
        115                 120                 125             
Phe Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Gly Ser 
    130                 135                 140                 
Thr Ser Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys Gly 
145                 150                 155                 160 
Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 
                165                 170                 175     
Asp Arg Val Thr Ile Thr Cys His Ala Ser Gln Asn Ile Tyr Val Trp 
            180                 185                 190         
Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 
        195                 200                 205             
Tyr Lys Ala Ser Asn Leu His Thr Gly Val Pro Ser Arg Phe Ser Gly 
    210                 215                 220                 
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 
225                 230                 235                 240 
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Gly Gln Thr Tyr Pro Tyr 
                245                 250                 255     
Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Glu Gln 
            260                 265                 270         
Lys Leu Ile Ser Glu Glu Asp Leu Phe Val Pro Val Phe Leu Pro Ala 
        275                 280                 285             
Lys Pro Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr 
    290                 295                 300                 
Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala 
305                 310                 315                 320 
Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile 
                325                 330                 335     
Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser 
            340                 345                 350         
Leu Val Ile Thr Leu Tyr Cys Asn His Arg Asn Arg Ser Lys Arg Ser 
        355                 360                 365             
Arg Leu Leu His Ser Asp Tyr Met Asn Met Thr Pro Arg Arg Pro Gly 
    370                 375                 380                 
Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala Pro Pro Arg Asp Phe Ala 
385                 390                 395                 400 
Ala Tyr Arg Ser Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala 
                405                 410                 415     
Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg 
            420                 425                 430         
Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu 
        435                 440                 445             
Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn 
    450                 455                 460                 
Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met 
465                 470                 475                 480 
Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly 
                485                 490                 495     
Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala 
            500                 505                 510         
Leu Pro Pro Arg 
        515     

<210> 44
<211> 514
<212> PRT
<213> Artificial Sequence


<220> 
<223> CD28_CAR_14 (including signal peptide)

<400> 44
Met Leu Leu Leu Val Thr Ser Leu Leu Leu Cys Glu Leu Pro His Pro 
1               5                   10                  15      
Ala Phe Leu Leu Ile Pro Val Lys Leu Gln Gln Ser Gly Ala Glu Leu 
            20                  25                  30          
Val Lys Pro Gly Ala Ser Val Arg Leu Ser Cys Lys Ala Ser Gly Tyr 
        35                  40                  45              
Thr Phe Thr Glu Tyr Ile Ile His Trp Ile Lys Leu Arg Ser Gly Gln 
    50                  55                  60                  
Gly Leu Glu Trp Ile Gly Trp Phe Tyr Pro Gly Ser Asn Asp Ile Gln 
65                  70                  75                  80  
Tyr Asn Ala Lys Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser 
                85                  90                  95      
Ser Ser Thr Val Tyr Met Glu Leu Thr Gly Leu Thr Ser Glu Asp Ser 
            100                 105                 110         
Ala Val Tyr Phe Cys Ala Arg Arg Asp Asp Phe Ser Gly Tyr Asp Ala 
        115                 120                 125             
Leu Pro Tyr Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Gly Ser 
    130                 135                 140                 
Thr Ser Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys Gly 
145                 150                 155                 160 
Asp Ile Gln Met Thr Gln Ser Pro Ala Ser Leu Ser Val Ser Val Gly 
                165                 170                 175     
Glu Thr Val Thr Ile Thr Cys Arg Thr Asn Glu Asn Ile Tyr Ser Asn 
            180                 185                 190         
Leu Ala Trp Tyr Gln Gln Lys Gln Gly Lys Ser Pro Gln Leu Leu Ile 
        195                 200                 205             
Tyr Ala Ala Thr His Leu Val Glu Gly Val Pro Ser Arg Phe Ser Gly 
    210                 215                 220                 
Ser Gly Ser Gly Thr Gln Tyr Ser Leu Lys Ile Thr Ser Leu Gln Ser 
225                 230                 235                 240 
Glu Asp Phe Gly Asn Tyr Tyr Cys Gln His Phe Trp Gly Thr Pro Cys 
                245                 250                 255     
Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Glu Gln Lys Leu 
            260                 265                 270         
Ile Ser Glu Glu Asp Leu Phe Val Pro Val Phe Leu Pro Ala Lys Pro 
        275                 280                 285             
Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala 
    290                 295                 300                 
Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly 
305                 310                 315                 320 
Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp Ile Tyr Ile 
                325                 330                 335     
Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu Leu Ser Leu Val 
            340                 345                 350         
Ile Thr Leu Tyr Cys Asn His Arg Asn Arg Ser Lys Arg Ser Arg Leu 
        355                 360                 365             
Leu His Ser Asp Tyr Met Asn Met Thr Pro Arg Arg Pro Gly Pro Thr 
    370                 375                 380                 
Arg Lys His Tyr Gln Pro Tyr Ala Pro Pro Arg Asp Phe Ala Ala Tyr 
385                 390                 395                 400 
Arg Ser Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln 
                405                 410                 415     
Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu 
            420                 425                 430         
Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly 
        435                 440                 445             
Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu 
    450                 455                 460                 
Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly 
465                 470                 475                 480 
Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser 
                485                 490                 495     
Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro 
            500                 505                 510         
Pro Arg 
        

<210> 45
<211> 1431
<212> DNA
<213> Artificial Sequence


<220> 
<223> CD28_CAR_1 encoding nucleotide sequence

<400> 45
atgttgctgt tagttacatc cttgctcctg tgcgaattgc cacatccagc attccttttg      60

atccccgata tccaaatgac acaaagcccg agttccctct ctgccagcgt aggagatcgc     120

gtaactataa cttgccacgc tagccaaaat atttacgtat ggctgaactg gtatcagcag     180

aaaccaggaa aagcaccgaa acttctcatt tataaagcca gcaatttgca cacaggggtt     240

ccgtcccgct tttccgggtc aggctcaggt acggacttca ccctgacgat ttccagcctt     300

caacccgagg actttgccac ttattattgt caacagggtc aaacataccc ctataccttc     360

ggcggtggga ctaaagtcga aattaagagg accgttggca gtactagcgg ctccggcaaa     420

ccagggtccg gcgaaggatc caccaaggga caagttcagc tggtacaaag tggcgcagag     480

gtaaagaaac ctggcgccag cgtgaaggtt tcctgtaagg catccggata cacctttaca     540

agctactata ttcattgggt tcggcaagct cctggccaag gtttggaatg gattgggtgc     600

atttaccccg gtaatgtcaa caccaattac aacgaaaagt ttaaggaccg ggccacgttg     660

actgttgata cgtctatctc cacagcgtac atggagctct ccagacttag atctgatgat     720

acagcagtgt atttctgtac gagatctcat tatggtcttg actggaactt tgacgtctgg     780

ggacagggaa cgaccgtcac tgtttctagt gaacagaaac tgatctccga ggaagatctc     840

ggaggaggcg ggtcaggtgg ggggggttct ggggggggcg ggtcaatata tatatgggca     900

ccactggccg ggacttgtgg ggtccttctc ctgtcactgg ttatcaccct ttactgcaac     960

cacaggaaca ggagtaagag gagcaggctc ctgcacagtg actacatgaa catgactccc    1020

cgccgccccg ggcccacccg caagcattac cagccctatg ccccaccacg cgacttcgca    1080

gcctatcgct ccagagtgaa gttcagcagg agcgcagacg cccccgcgta ccagcagggc    1140

cagaaccagc tctataacga gctcaatcta ggacgaagag aggagtacga tgttttggac    1200

aagagacgtg gccgggaccc tgagatgggg ggaaagccga gaaggaagaa ccctcaggaa    1260

ggcctgtaca atgaactgca gaaagataag atggcggagg cctacagtga gattgggatg    1320

aaaggcgagc gccggagggg caaggggcac gatggccttt accagggtct cagtacagcc    1380

accaaggaca cctacgacgc ccttcacatg caggccctgc cccctcgcta a             1431


<210> 46
<211> 1551
<212> DNA
<213> Artificial Sequence


<220> 
<223> CD28_CAR_2 encoding nucleotide sequence

<400> 46
atgttgctgt tagttacatc cttgctcctg tgcgaactcc cacatcccgc ttttctcctg      60

atcccagata ttcaaatgac ccaaagccct tcttctctta gcgcaagcgt gggtgatcgg     120

gtcacaatta cgtgccacgc gtctcaaaat atatacgtct ggctgaactg gtatcagcag     180

aaaccaggaa aagcaccaaa actgcttatc tataaagcaa gcaatctcca cactggggta     240

cctagccgct tttccggcag cggctcaggc acagacttca ccttgactat ttccagtctg     300

caacctgagg actttgctac atattattgt caacagggtc aaacataccc ctatactttc     360

ggggggggga ctaaagtgga aattaagcgc accgtaggct ccacttcagg gagcggcaaa     420

cctgggagcg gcgaaggcag cactaaggga caagtccagc ttgtgcaatc tggagcagag     480

gtcaagaaac cgggcgcctc agttaaggtc tcctgtaagg ctagcggata cacatttact     540

tcctactata tacattgggt gcgacaagct ccgggacaag ggcttgaatg gattggttgc     600

atttaccctg gaaatgtcaa caccaattac aacgaaaagt ttaaggacag agccaccttg     660

acggtggata cgtcaatatc cactgcttac atggagctga gcagattgag gtccgatgat     720

acggctgttt acttttgtac taggagtcat tatgggcttg actggaactt tgacgtatgg     780

ggacagggta ctaccgtcac agttagttca gaacagaaac tgatcagtga ggaagatctg     840

tttgtccctg tgtttcttcc cgccaaacct actaccactc ctgcacccag gccacccacc     900

ccagccccaa caattgcctc tcaacctctc tctctccgac ctgaagcctg taggccggct     960

gctggcggag ctgtacatac acgaggtctc gattttgctt gcgacattta tatttgggcc    1020

ccactcgccg ggacttgtgg ggtccttctc ctgtcactgg ttatcaccct ttactgcaac    1080

cacaggaaca ggagtaagag gagcaggctc ctgcacagtg actacatgaa catgactccc    1140

cgccgccccg ggcccacccg caagcattac cagccctatg ccccaccacg cgacttcgca    1200

gcctatcgct ccagagtgaa gttcagcagg agcgcagacg cccccgcgta ccagcagggc    1260

cagaaccagc tctataacga gctcaatcta ggacgaagag aggagtacga tgttttggac    1320

aagagacgtg gccgggaccc tgagatgggg ggaaagccga gaaggaagaa ccctcaggaa    1380

ggcctgtaca atgaactgca gaaagataag atggcggagg cctacagtga gattgggatg    1440

aaaggcgagc gccggagggg caaggggcac gatggccttt accagggtct cagtacagcc    1500

accaaggaca cctacgacgc ccttcacatg caggccctgc cccctcgcta a             1551


<210> 47
<211> 1431
<212> DNA
<213> Artificial Sequence


<220> 
<223> CD28_CAR_11 encoding nucleotide sequence

<400> 47
atgttgctgt tagttacatc cttgctcctg tgcgaattgc cacatccagc tttccttctg      60

atcccacaag tccagctggt gcaaagtggc gccgaggtta agaaacctgg cgcatccgtg     120

aaggtaagct gtaaggctag cggctacacc tttacaagct actatattca ttgggtgaga     180

caagcaccag gacaaggact cgaatggata ggctgcattt accctggcaa tgtaaacacg     240

aattacaacg aaaagtttaa ggaccgagcc accctgacag ttgatactag catcagtact     300

gcgtacatgg agcttagccg tcttaggagc gatgatactg cggtttactt ttgtaccagg     360

tcacattatg ggcttgactg gaactttgac gtctggggac agggaacaac tgtcaccgtt     420

tcttccggtt ccacttcagg ctccggtaaa ccagggtctg gcgaaggcag caccaagggc     480

gatatccaaa tgacacaatc tccctcatca ttgagtgcaa gtgtgggtga tcgcgtgact     540

attacatgcc acgcctctca aaatatttac gtgtggctga actggtatca gcagaaaccc     600

gggaaagcac cgaaactgct catttataaa gcatctaatt tgcacaccgg tgtaccctca     660

agatttagcg gatcaggctc aggcacagac ttcacactga caatttcaag ccttcaaccc     720

gaggactttg ccacatatta ttgtcaacag gggcaaacat acccttatac tttcggcggc     780

gggacgaaag tggaaattaa gcgcaccgtt gaacagaaac tgatctccga ggaagatctc     840

ggcggggggg gatccggtgg aggagggtcc ggaggaggtg gttccattta tatttgggca     900

ccactggccg ggacttgtgg ggtccttctc ctgtcactgg ttatcaccct ttactgcaac     960

cacaggaaca ggagtaagag gagcaggctc ctgcacagtg actacatgaa catgactccc    1020

cgccgccccg ggcccacccg caagcattac cagccctatg ccccaccacg cgacttcgca    1080

gcctatcgct ccagagtgaa gttcagcagg agcgcagacg cccccgcgta ccagcagggc    1140

cagaaccagc tctataacga gctcaatcta ggacgaagag aggagtacga tgttttggac    1200

aagagacgtg gccgggaccc tgagatgggg ggaaagccga gaaggaagaa ccctcaggaa    1260

ggcctgtaca atgaactgca gaaagataag atggcggagg cctacagtga gattgggatg    1320

aaaggcgagc gccggagggg caaggggcac gatggccttt accagggtct cagtacagcc    1380

accaaggaca cctacgacgc ccttcacatg caggccctgc cccctcgcta a             1431


<210> 48
<211> 1551
<212> DNA
<213> Artificial Sequence


<220> 
<223> CD28_CAR_12 encoding nucleotide sequence

<400> 48
atgttgctgt tagttacatc cttgctcctg tgcgaactgc cccatccagc gttcctgctg      60

atcccccaag tccagctggt gcaaagtggc gccgaggtca agaagccagg agccagcgtt     120

aaggtatctt gtaaggcatc aggctacacc tttacatctt actatattca ttgggtgaga     180

caagctccgg gacaagggtt ggaatggata gggtgcatat accctggtaa tgtcaacact     240

aattacaacg aaaagtttaa ggaccgggcc acactcactg tggatacgtc aatcagtacg     300

gcatacatgg agctgagccg gctccggagc gatgatacag cggtgtactt ttgtacccgt     360

tcccattatg ggctcgactg gaactttgac gtatggggcc aggggacaac cgttaccgtc     420

agtagtggca gcacctcagg gagcggtaaa ccaggatctg gcgaaggctc cactaaggga     480

gatattcaaa tgacgcaatc accctcatca ctgtctgcca gtgtgggaga tcgtgtcacg     540

attacttgcc acgcttctca aaatatttac gtgtggttga actggtatca gcaaaagccc     600

gggaaagcac caaaactctt gatctataaa gcaagcaatc tccacacagg ggtcccgagc     660

cgattttctg ggtccggcag tgggacagac ttcacactga cgatttcatc actccaaccg     720

gaagactttg ccacttatta ttgtcaacag ggccaaacct acccctatac tttcggcggc     780

gggaccaaag tcgaaataaa gcggaccgtt gaacagaaat tgatctccga ggaagatctg     840

tttgtcccag tgtttcttcc tgctaaacca actactactc ccgcccccag acccccgact     900

cccgccccaa ctattgcttc tcaacctttg agtctcagac ccgaagcctg taggccggcc     960

gccggtgggg ccgtccatac tcgcggtttg gattttgcat gcgacattta tatatgggcc    1020

cctctggccg ggacttgtgg ggtccttctc ctgtcactgg ttatcaccct ttactgcaac    1080

cacaggaaca ggagtaagag gagcaggctc ctgcacagtg actacatgaa catgactccc    1140

cgccgccccg ggcccacccg caagcattac cagccctatg ccccaccacg cgacttcgca    1200

gcctatcgct ccagagtgaa gttcagcagg agcgcagacg cccccgcgta ccagcagggc    1260

cagaaccagc tctataacga gctcaatcta ggacgaagag aggagtacga tgttttggac    1320

aagagacgtg gccgggaccc tgagatgggg ggaaagccga gaaggaagaa ccctcaggaa    1380

ggcctgtaca atgaactgca gaaagataag atggcggagg cctacagtga gattgggatg    1440

aaaggcgagc gccggagggg caaggggcac gatggccttt accagggtct cagtacagcc    1500

accaaggaca cctacgacgc ccttcacatg caggccctgc cccctcgcta a             1551


<210> 49
<211> 1545
<212> DNA
<213> Artificial Sequence


<220> 
<223> CD28_CAR_14 encoding nucleotide sequence

<400> 49
atgttgctgt tagttacatc cttgctcctg tgcgaactgc cccatcccgc gttccttctc      60

atcccagtaa aacttcaaca atctggtgct gaattggtca aaccaggggc ttccgtacgg     120

ctttcttgta aagcttccgg ttatactttt acggaatata ttatccattg gataaaactt     180

cgtagtggtc agggactcga gtggattggt tggttttatc ccggatccaa tgatatacaa     240

tataatgcaa aatttaaagg gaaagctact ctcactgcag ataagtcaag ttccacagtc     300

tacatggaac ttactggact cacatccgaa gattctgctg tctacttttg tgcacggcgc     360

gatgatttct ctggatatga tgctcttcca tattggggac aaggaacaat ggtaacagtt     420

tcctctggta gcacttcagg atctggcaaa ccgggttctg gggaaggttc caccaagggc     480

gatatacaaa tgacacaaag tccagctagt ctttccgtct ccgtcggaga aacggtcacg     540

attacgtgtc gaaccaatga aaatatatat tcaaatctcg catggtatca acaaaagcaa     600

ggaaaatctc cacaacttct gatatatgcg gcgactcatc tcgttgaagg ggtaccgtct     660

cggttttcag ggtccggatc cggaacacaa tatagtctca aaataacaag tctccaatct     720

gaggactttg ggaattatta ttgtcaacat ttctggggta ctccatgtac atttggagga     780

ggaactaaac ttgaaataaa acgagaacag aaattgatat ccgaggaaga tcttttcgtt     840

cccgtgtttc tccctgcaaa accgacaact actcctgctc caaggccgcc cactcccgcc     900

cctactatag catcccaacc acttagtttg agacctgaag cctgtaggcc cgccgcaggt     960

ggagctgtcc atacacgcgg cctcgatttt gcttgcgaca tttatatttg ggcccctctg    1020

gccgggactt gtggggtcct tctcctgtca ctggttatca ccctttactg caaccacagg    1080

aacaggagta agaggagcag gctcctgcac agtgactaca tgaacatgac tccccgccgc    1140

cccgggccca cccgcaagca ttaccagccc tatgccccac cacgcgactt cgcagcctat    1200

cgctccagag tgaagttcag caggagcgca gacgcccccg cgtaccagca gggccagaac    1260

cagctctata acgagctcaa tctaggacga agagaggagt acgatgtttt ggacaagaga    1320

cgtggccggg accctgagat ggggggaaag ccgagaagga agaaccctca ggaaggcctg    1380

tacaatgaac tgcagaaaga taagatggcg gaggcctaca gtgagattgg gatgaaaggc    1440

gagcgccgga ggggcaaggg gcacgatggc ctttaccagg gtctcagtac agccaccaag    1500

gacacctacg acgcccttca catgcaggcc ctgccccctc gctaa                    1545


<210> 50
<211> 20
<212> RNA
<213> Artificial Sequence


<220> 
<223> spacer sequence of crRNA (targeting exon 2 of human CD28-gene)

<400> 50
gcuuguagcg uacgacaaug                                                20


<210> 51
<211> 36
<212> RNA
<213> Artificial Sequence


<220> 
<223> crRNA (targeting exon 2 of human CD28-gene)

<400> 51
gcuuguagcg uacgacaaug guuuuagagc uaugcu                               36


<210> 52
<211> 4
<212> PRT
<213> Artificial Sequence


<220> 
<223> sequence motif in ITAM

<220> 
<221> VARIANT
<222> 2
<223> any amino acid

<220> 
<221> VARIANT
<222> 3
<223> any amino acid

<220> 
<221> VARIANT
<222> 4
<223> Leu or Ile

<400> 52
Tyr Xaa Xaa Leu 
1               

<210> 53
<211> 14
<212> PRT
<213> Artificial Sequence


<220> 
<223> ITAM conserved motif

<220> 
<221> VARIANT
<222> 2
<223> any amino acid

<220> 
<221> VARIANT
<222> 3
<223> any amino acid

<220> 
<221> VARIANT
<222> 4
<223> Leu or Ile

<220> 
<221> VARIANT
<222> 5
<223> any amino acid

<220> 
<221> VARIANT
<222> 6
<223> any amino acid

<220> 
<221> VARIANT
<222> 7
<223> any amino acid

<220> 
<221> VARIANT
<222> 8
<223> any amino acid

<220> 
<221> VARIANT
<222> 9
<223> any amino acid

<220> 
<221> VARIANT
<222> 10
<223> Xaa or XaaXaa or XaaXaaXaa, wherein Xaa can be any amino acid

<220> 
<221> VARIANT
<222> 12
<223> any amino acid

<220> 
<221> VARIANT
<222> 13
<223> any amino acid

<220> 
<221> VARIANT
<222> 14
<223> Leu or Ile

<400> 53
Tyr Xaa Xaa Leu Xaa Xaa Xaa Xaa Xaa Xaa Tyr Xaa Xaa Leu 
1               5                   10                  

<210> 54
<211> 100
<212> DNA
<213> Artificial Sequence


<220> 
<223> 100 bp upstream of ATG start codon of pMP71

<400> 54
tcgagtaata cgactcacta tagggagacc caagctggct aggtaagctt gatcaacaag     60

tttgtacaaa aaagcaggct ccgcggccgc ccccttcacc                          100


<210> 55
<211> 356
<212> PRT
<213> Artificial Sequence


<220> 
<223> truncated human epidermal growth factor receptor (EGFRt)
      epitope-tag (including signal peptide (amino acids 1-18))

<400> 55
Leu Leu Leu Val Thr Ser Leu Leu Leu Cys Glu Leu Pro His Pro Ala 
1               5                   10                  15      
Phe Leu Leu Ile Pro Arg Lys Val Cys Asn Gly Ile Gly Ile Gly Glu 
            20                  25                  30          
Phe Lys Asp Ser Leu Ser Ile Asn Ala Thr Asn Ile Lys His Phe Lys 
        35                  40                  45              
Asn Cys Thr Ser Ile Ser Gly Asp Leu His Ile Leu Pro Val Ala Phe 
    50                  55                  60                  
Arg Gly Asp Ser Phe Thr His Thr Pro Pro Leu Asp Pro Gln Glu Leu 
65                  70                  75                  80  
Asp Ile Leu Lys Thr Val Lys Glu Ile Thr Gly Phe Leu Leu Ile Gln 
                85                  90                  95      
Ala Trp Pro Glu Asn Arg Thr Asp Leu His Ala Phe Glu Asn Leu Glu 
            100                 105                 110         
Ile Ile Arg Gly Arg Thr Lys Gln His Gly Gln Phe Ser Leu Ala Val 
        115                 120                 125             
Val Ser Leu Asn Ile Thr Ser Leu Gly Leu Arg Ser Leu Lys Glu Ile 
    130                 135                 140                 
Ser Asp Gly Asp Val Ile Ile Ser Gly Asn Lys Asn Leu Cys Tyr Ala 
145                 150                 155                 160 
Asn Thr Ile Asn Trp Lys Lys Leu Phe Gly Thr Ser Gly Gln Lys Thr 
                165                 170                 175     
Lys Ile Ile Ser Asn Arg Gly Glu Asn Ser Cys Lys Ala Thr Gly Gln 
            180                 185                 190         
Val Cys His Ala Leu Cys Ser Pro Glu Gly Cys Trp Gly Pro Glu Pro 
        195                 200                 205             
Arg Asp Cys Val Ser Cys Arg Asn Val Ser Arg Gly Arg Glu Cys Val 
    210                 215                 220                 
Asp Lys Cys Asn Leu Leu Glu Gly Glu Pro Arg Glu Phe Val Glu Asn 
225                 230                 235                 240 
Ser Glu Cys Ile Gln Cys His Pro Glu Cys Leu Pro Gln Ala Met Asn 
                245                 250                 255     
Ile Thr Cys Thr Gly Arg Gly Pro Asp Asn Cys Ile Gln Cys Ala His 
            260                 265                 270         
Tyr Ile Asp Gly Pro His Cys Val Lys Thr Cys Pro Ala Gly Val Met 
        275                 280                 285             
Gly Glu Asn Asn Thr Leu Val Trp Lys Tyr Ala Asp Ala Gly His Val 
    290                 295                 300                 
Cys His Leu Cys His Pro Asn Cys Thr Tyr Gly Cys Thr Gly Pro Gly 
305                 310                 315                 320 
Leu Glu Gly Cys Pro Thr Asn Gly Pro Lys Ile Pro Ser Ile Ala Thr 
                325                 330                 335     
Gly Met Val Gly Ala Leu Leu Leu Leu Leu Val Val Ala Leu Gly Ile 
            340                 345                 350         
Gly Leu Phe Met 
        355     

<210> 56
<211> 21
<212> PRT
<213> Artificial Sequence


<220> 
<223> T2A linker peptide

<400> 56
Gly Ser Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly Asp Val Glu 
1               5                   10                  15      
Glu Asn Pro Gly Pro 
            20      

<210> 57
<211> 123
<212> PRT
<213> Artificial Sequence


<220> 
<223> VH of an anti-CD7 single-chain variable fragment (scFv)

<400> 57
Gln Val Gln Leu Gln Glu Ser Gly Ala Glu Leu Val Lys Pro Gly Ala 
1               5                   10                  15      
Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 
            20                  25                  30          
Trp Met His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 
        35                  40                  45              
Gly Lys Ile Asn Pro Ser Asn Gly Arg Thr Asn Tyr Asn Glu Lys Phe 
    50                  55                  60                  
Lys Ser Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr 
65                  70                  75                  80  
Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 
                85                  90                  95      
Ala Arg Gly Gly Val Tyr Tyr Asp Leu Tyr Tyr Tyr Ala Leu Asp Tyr 
            100                 105                 110         
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 
        115                 120             

<210> 58
<211> 108
<212> PRT
<213> Artificial Sequence


<220> 
<223> VL of an anti-CD7 single-chain variable fragment (scFv)

<400> 58
Asp Ile Glu Leu Thr Gln Ser Pro Ala Thr Leu Ser Val Thr Pro Gly 
1               5                   10                  15      
Asp Ser Val Ser Leu Ser Cys Arg Ala Ser Gln Ser Ile Ser Asn Asn 
            20                  25                  30          
Leu His Trp Tyr Gln Gln Lys Ser His Glu Ser Pro Arg Leu Leu Ile 
        35                  40                  45              
Lys Ser Ala Ser Gln Ser Ile Ser Gly Ile Pro Ser Arg Phe Ser Gly 
    50                  55                  60                  
Ser Gly Ser Gly Thr Asp Phe Thr Leu Ser Ile Asn Ser Val Glu Thr 
65                  70                  75                  80  
Glu Asp Phe Gly Met Tyr Phe Cys Gln Gln Ser Asn Ser Trp Pro Tyr 
                85                  90                  95      
Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg 
            100                 105             

