
                                SEQUENCE LISTING

<110> Sachs, David H.
      Huang, Christene A.
      The General Hospital Corporation

<120> Recombinant Cytotoxic
  T-Lymphocyte-Associated Protein 4 (CTLA4)
  

<130> 29539-0061WO1

<150> US 61/617,345       
<151> 2012-03-29  

<160> 31

<170> FastSEQ for Windows Version 4.0

<210> 1
<211> 174
<212> PRT
<213> Homo sapien

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


<210> 2
<211> 528
<212> PRT
<213> Artificial Sequence

<220> 
<223> DT390-pCTLA4-Non-N-Gly-6xHis

<400> 2
Ala Gly Ala Asp Asp Val Val Asp Ser Ser Lys Ser Phe Val Met Glu
 1               5                  10                  15      
Asn Phe Ala Ser Tyr His Gly Thr Lys Pro Gly Tyr Val Asp Ser Ile
            20                  25                  30          
Gln Lys Gly Ile Gln Lys Pro Lys Ser Gly Thr Gln Gly Asn Tyr Asp
        35                  40                  45              
Asp Asp Trp Lys Gly Phe Tyr Ser Thr Asp Asn Lys Tyr Asp Ala Ala
    50                  55                  60                  
Gly Tyr Ser Val Asp Asn Glu Asn Pro Leu Ser Gly Lys Ala Gly Gly
65                  70                  75                  80  
Val Val Lys Val Thr Tyr Pro Gly Leu Thr Lys Val Leu Ala Leu Lys
                85                  90                  95      
Val Asp Asn Ala Glu Thr Ile Lys Lys Glu Leu Gly Leu Ser Leu Thr
            100                 105                 110         
Glu Pro Leu Met Glu Gln Val Gly Thr Glu Glu Phe Ile Lys Arg Phe
        115                 120                 125             
Gly Asp Gly Ala Ser Arg Val Val Leu Ser Leu Pro Phe Ala Glu Gly
    130                 135                 140                 
Ser Ser Ser Val Glu Tyr Ile Asn Asn Trp Glu Gln Ala Lys Ala Leu
145                 150                 155                 160 
Ser Val Glu Leu Glu Ile Asn Phe Glu Thr Arg Gly Lys Arg Gly Gln
                165                 170                 175     
Asp Ala Met Tyr Glu Tyr Met Ala Gln Ala Cys Ala Gly Asn Arg Val
            180                 185                 190         
Arg Arg Ser Val Gly Ser Ser Leu Ser Cys Ile Asn Leu Asp Trp Asp
        195                 200                 205             
Val Ile Arg Asp Lys Thr Lys Thr Lys Ile Glu Ser Leu Lys Glu His
    210                 215                 220                 
Gly Pro Ile Lys Asn Lys Met Ser Glu Ser Pro Ala Lys Thr Val Ser
225                 230                 235                 240 
Glu Glu Lys Ala Lys Gln Tyr Leu Glu Glu Phe His Gln Thr Ala Leu
                245                 250                 255     
Glu His Pro Glu Leu Ser Glu Leu Lys Thr Val Thr Gly Thr Asn Pro
            260                 265                 270         
Val Phe Ala Gly Ala Asn Tyr Ala Ala Trp Ala Val Asn Val Ala Gln
        275                 280                 285             
Val Ile Asp Ser Glu Thr Ala Asp Asn Leu Glu Lys Thr Thr Ala Ala
    290                 295                 300                 
Leu Ser Ile Leu Pro Gly Ile Gly Ser Val Met Gly Ile Ala Asp Gly
305                 310                 315                 320 
Ala Val His His Asn Thr Glu Glu Ile Val Ala Gln Ser Ile Ala Leu
                325                 330                 335     
Ser Ser Leu Met Val Ala Gln Ala Ile Pro Leu Val Gly Glu Leu Val
            340                 345                 350         
Asp Ile Gly Phe Ala Ala Tyr Asn Phe Val Glu Ser Ile Ile Asn Leu
        355                 360                 365             
Phe Gln Val Val His Asn Ser Tyr Asn Arg Pro Ala Tyr Ser Pro Gly
    370                 375                 380                 
His Lys Thr Gln Pro Phe Leu Pro Trp Gly Gly Gly Gly Ser Met His
385                 390                 395                 400 
Val Ala Gln Pro Ala Val Val Leu Ala Asn Ser Arg Gly Val Ala Ser
                405                 410                 415     
Phe Val Cys Glu Tyr Gly Ser Ala Gly Lys Ala Ala Glu Val Arg Val
            420                 425                 430         
Thr Val Leu Arg Arg Ala Gly Ser Gln Met Thr Glu Val Cys Ala Ala
        435                 440                 445             
Thr Tyr Thr Val Glu Asp Glu Leu Thr Phe Leu Asp Asp Ser Thr Cys
    450                 455                 460                 
Thr Gly Thr Ser Thr Glu Asn Lys Val Ala Leu Thr Ile Gln Gly Leu
465                 470                 475                 480 
Arg Ala Val Asp Thr Gly Leu Tyr Ile Cys Lys Val Glu Leu Leu Tyr
                485                 490                 495     
Pro Pro Pro Tyr Tyr Val Gly Met Gly Ala Gly Thr Gln Ile Tyr Val
            500                 505                 510         
Ile Asp Pro Glu Pro Cys Pro Asp Ser Asp His His His His His His
        515                 520                 525             


<210> 3
<211> 667
<212> PRT
<213> Artificial Sequence

<220> 
<223> DT390-bi-pCTLA-4-Non-N-Gly-6xHis

<400> 3
Ala Gly Ala Asp Asp Val Val Asp Ser Ser Lys Ser Phe Val Met Glu
 1               5                  10                  15      
Asn Phe Ala Ser Tyr His Gly Thr Lys Pro Gly Tyr Val Asp Ser Ile
            20                  25                  30          
Gln Lys Gly Ile Gln Lys Pro Lys Ser Gly Thr Gln Gly Asn Tyr Asp
        35                  40                  45              
Asp Asp Trp Lys Gly Phe Tyr Ser Thr Asp Asn Lys Tyr Asp Ala Ala
    50                  55                  60                  
Gly Tyr Ser Val Asp Asn Glu Asn Pro Leu Ser Gly Lys Ala Gly Gly
65                  70                  75                  80  
Val Val Lys Val Thr Tyr Pro Gly Leu Thr Lys Val Leu Ala Leu Lys
                85                  90                  95      
Val Asp Asn Ala Glu Thr Ile Lys Lys Glu Leu Gly Leu Ser Leu Thr
            100                 105                 110         
Glu Pro Leu Met Glu Gln Val Gly Thr Glu Glu Phe Ile Lys Arg Phe
        115                 120                 125             
Gly Asp Gly Ala Ser Arg Val Val Leu Ser Leu Pro Phe Ala Glu Gly
    130                 135                 140                 
Ser Ser Ser Val Glu Tyr Ile Asn Asn Trp Glu Gln Ala Lys Ala Leu
145                 150                 155                 160 
Ser Val Glu Leu Glu Ile Asn Phe Glu Thr Arg Gly Lys Arg Gly Gln
                165                 170                 175     
Asp Ala Met Tyr Glu Tyr Met Ala Gln Ala Cys Ala Gly Asn Arg Val
            180                 185                 190         
Arg Arg Ser Val Gly Ser Ser Leu Ser Cys Ile Asn Leu Asp Trp Asp
        195                 200                 205             
Val Ile Arg Asp Lys Thr Lys Thr Lys Ile Glu Ser Leu Lys Glu His
    210                 215                 220                 
Gly Pro Ile Lys Asn Lys Met Ser Glu Ser Pro Ala Lys Thr Val Ser
225                 230                 235                 240 
Glu Glu Lys Ala Lys Gln Tyr Leu Glu Glu Phe His Gln Thr Ala Leu
                245                 250                 255     
Glu His Pro Glu Leu Ser Glu Leu Lys Thr Val Thr Gly Thr Asn Pro
            260                 265                 270         
Val Phe Ala Gly Ala Asn Tyr Ala Ala Trp Ala Val Asn Val Ala Gln
        275                 280                 285             
Val Ile Asp Ser Glu Thr Ala Asp Asn Leu Glu Lys Thr Thr Ala Ala
    290                 295                 300                 
Leu Ser Ile Leu Pro Gly Ile Gly Ser Val Met Gly Ile Ala Asp Gly
305                 310                 315                 320 
Ala Val His His Asn Thr Glu Glu Ile Val Ala Gln Ser Ile Ala Leu
                325                 330                 335     
Ser Ser Leu Met Val Ala Gln Ala Ile Pro Leu Val Gly Glu Leu Val
            340                 345                 350         
Asp Ile Gly Phe Ala Ala Tyr Asn Phe Val Glu Ser Ile Ile Asn Leu
        355                 360                 365             
Phe Gln Val Val His Asn Ser Tyr Asn Arg Pro Ala Tyr Ser Pro Gly
    370                 375                 380                 
His Lys Thr Gln Pro Phe Leu Pro Trp Gly Gly Gly Gly Ser Met His
385                 390                 395                 400 
Val Ala Gln Pro Ala Val Val Leu Ala Asn Ser Arg Gly Val Ala Ser
                405                 410                 415     
Phe Val Cys Glu Tyr Gly Ser Ala Gly Lys Ala Ala Glu Val Arg Val
            420                 425                 430         
Thr Val Leu Arg Arg Ala Gly Ser Gln Met Thr Glu Val Cys Ala Ala
        435                 440                 445             
Thr Tyr Thr Val Glu Asp Glu Leu Thr Phe Leu Asp Asp Ser Thr Cys
    450                 455                 460                 
Thr Gly Thr Ser Thr Glu Asn Lys Val Ala Leu Thr Ile Gln Gly Leu
465                 470                 475                 480 
Arg Ala Val Asp Thr Gly Leu Tyr Ile Cys Lys Val Glu Leu Leu Tyr
                485                 490                 495     
Pro Pro Pro Tyr Tyr Val Gly Met Gly Ala Gly Thr Gln Ile Tyr Val
            500                 505                 510         
Ile Asp Pro Glu Pro Cys Pro Asp Ser Asp Gly Gly Gly Gly Ser Gly
        515                 520                 525             
Gly Gly Gly Ser Gly Gly Gly Gly Ser Met His Val Ala Gln Pro Ala
    530                 535                 540                 
Val Val Leu Ala Asn Ser Arg Gly Val Ala Ser Phe Val Cys Glu Tyr
545                 550                 555                 560 
Gly Ser Ala Gly Lys Ala Ala Glu Val Arg Val Thr Val Leu Arg Arg
                565                 570                 575     
Ala Gly Ser Gln Met Thr Glu Val Cys Ala Ala Thr Tyr Thr Val Glu
            580                 585                 590         
Asp Glu Leu Thr Phe Leu Asp Asp Ser Thr Cys Thr Gly Thr Ser Thr
        595                 600                 605             
Glu Asn Lys Val Ala Leu Thr Ile Gln Gly Leu Arg Ala Val Asp Thr
    610                 615                 620                 
Gly Leu Tyr Ile Cys Lys Val Glu Leu Leu Tyr Pro Pro Pro Tyr Tyr
625                 630                 635                 640 
Val Gly Met Gly Ala Gly Thr Gln Ile Tyr Val Ile Asp Pro Glu Pro
                645                 650                 655     
Cys Pro Asp Ser Asp His His His His His His
            660                 665         


<210> 4
<211> 528
<212> PRT
<213> Artificial Sequence

<220> 
<223> DT390-pCTLA-4-Gly-6xHis

<400> 4
Ala Gly Ala Asp Asp Val Val Asp Ser Ser Lys Ser Phe Val Met Glu
 1               5                  10                  15      
Asn Phe Ala Ser Tyr His Gly Thr Lys Pro Gly Tyr Val Asp Ser Ile
            20                  25                  30          
Gln Lys Gly Ile Gln Lys Pro Lys Ser Gly Thr Gln Gly Asn Tyr Asp
        35                  40                  45              
Asp Asp Trp Lys Gly Phe Tyr Ser Thr Asp Asn Lys Tyr Asp Ala Ala
    50                  55                  60                  
Gly Tyr Ser Val Asp Asn Glu Asn Pro Leu Ser Gly Lys Ala Gly Gly
65                  70                  75                  80  
Val Val Lys Val Thr Tyr Pro Gly Leu Thr Lys Val Leu Ala Leu Lys
                85                  90                  95      
Val Asp Asn Ala Glu Thr Ile Lys Lys Glu Leu Gly Leu Ser Leu Thr
            100                 105                 110         
Glu Pro Leu Met Glu Gln Val Gly Thr Glu Glu Phe Ile Lys Arg Phe
        115                 120                 125             
Gly Asp Gly Ala Ser Arg Val Val Leu Ser Leu Pro Phe Ala Glu Gly
    130                 135                 140                 
Ser Ser Ser Val Glu Tyr Ile Asn Asn Trp Glu Gln Ala Lys Ala Leu
145                 150                 155                 160 
Ser Val Glu Leu Glu Ile Asn Phe Glu Thr Arg Gly Lys Arg Gly Gln
                165                 170                 175     
Asp Ala Met Tyr Glu Tyr Met Ala Gln Ala Cys Ala Gly Asn Arg Val
            180                 185                 190         
Arg Arg Ser Val Gly Ser Ser Leu Ser Cys Ile Asn Leu Asp Trp Asp
        195                 200                 205             
Val Ile Arg Asp Lys Thr Lys Thr Lys Ile Glu Ser Leu Lys Glu His
    210                 215                 220                 
Gly Pro Ile Lys Asn Lys Met Ser Glu Ser Pro Ala Lys Thr Val Ser
225                 230                 235                 240 
Glu Glu Lys Ala Lys Gln Tyr Leu Glu Glu Phe His Gln Thr Ala Leu
                245                 250                 255     
Glu His Pro Glu Leu Ser Glu Leu Lys Thr Val Thr Gly Thr Asn Pro
            260                 265                 270         
Val Phe Ala Gly Ala Asn Tyr Ala Ala Trp Ala Val Asn Val Ala Gln
        275                 280                 285             
Val Ile Asp Ser Glu Thr Ala Asp Asn Leu Glu Lys Thr Thr Ala Ala
    290                 295                 300                 
Leu Ser Ile Leu Pro Gly Ile Gly Ser Val Met Gly Ile Ala Asp Gly
305                 310                 315                 320 
Ala Val His His Asn Thr Glu Glu Ile Val Ala Gln Ser Ile Ala Leu
                325                 330                 335     
Ser Ser Leu Met Val Ala Gln Ala Ile Pro Leu Val Gly Glu Leu Val
            340                 345                 350         
Asp Ile Gly Phe Ala Ala Tyr Asn Phe Val Glu Ser Ile Ile Asn Leu
        355                 360                 365             
Phe Gln Val Val His Asn Ser Tyr Asn Arg Pro Ala Tyr Ser Pro Gly
    370                 375                 380                 
His Lys Thr Gln Pro Phe Leu Pro Trp Gly Gly Gly Gly Ser Met His
385                 390                 395                 400 
Val Ala Gln Pro Ala Val Val Leu Ala Asn Ser Arg Gly Val Ala Ser
                405                 410                 415     
Phe Val Cys Glu Tyr Gly Ser Ala Gly Lys Ala Ala Glu Val Arg Val
            420                 425                 430         
Thr Val Leu Arg Arg Ala Gly Ser Gln Met Thr Glu Val Cys Ala Ala
        435                 440                 445             
Thr Tyr Thr Val Glu Asp Glu Leu Thr Phe Leu Asp Asp Ser Thr Cys
    450                 455                 460                 
Thr Gly Thr Ser Thr Glu Asn Lys Val Asn Leu Thr Ile Gln Gly Leu
465                 470                 475                 480 
Arg Ala Val Asp Thr Gly Leu Tyr Ile Cys Lys Val Glu Leu Leu Tyr
                485                 490                 495     
Pro Pro Pro Tyr Tyr Val Gly Met Gly Asn Gly Thr Gln Ile Tyr Val
            500                 505                 510         
Ile Asp Pro Glu Pro Cys Pro Asp Ser Asp His His His His His His
        515                 520                 525             


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

<220> 
<223> synthetically generated peptides

<400> 5
Asp Tyr Lys Asp Asp Asp Asp Lys
 1               5              


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

<220> 
<223> synthetically generated peptides

<400> 6
Tyr Pro Tyr Asp Val Pro
 1               5      


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

<220> 
<223> synthetically generated peptides

<400> 7
Ile Leu Lys Lys Ala Thr Ala Tyr Ile Leu
 1               5                  10  


<210> 8
<211> 10
<212> PRT
<213> Artificial Sequence

<220> 
<223> synthetically generated peptides

<400> 8
Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu
 1               5                  10  


<210> 9
<211> 57
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetically generated PCR primers

<400> 9
ccgctcgaga agagagaggc tgaagctatg cacgttgctc aaccagctgt tgtcttg    57

<210> 10
<211> 63
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetically generated PCR primers

<400> 10
accgtactca caaacgaaag aagcaacacc tctagagtta gccaagacaa cagctggttg 60
agc                                                               63

<210> 11
<211> 63
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetically generated PCR primers

<400> 11
tctttcgttt gtgagtacgg ttctgctggt aaggctgctg aggttagagt tactgttttg 60
aga                                                               63

<210> 12
<211> 63
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetically generated PCR primers

<400> 12
gtaagtagca gcacaaacct cagtcatttg agaaccagct cttctcaaaa cagtaactct 60
aac                                                               63

<210> 13
<211> 63
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetically generated PCR primers

<400> 13
gaggtttgtg ctgctactta cactgttgag gacgagttga ctttcttgga cgactctact 60
tgt                                                               63

<210> 14
<211> 63
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetically generated PCR primers

<400> 14
ttgaatagtc aagttaacct tgttctcagt agaagtacca gtacaagtag agtcgtccaa 60
gaa                                                               63

<210> 15
<211> 63
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetically generated PCR primers

<400> 15
aaggttaact tgactattca aggtttgaga gctgtcgaca ccggtttgta catctgtaag 60
gtc                                                               63

<210> 16
<211> 63
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetically generated PCR primers

<400> 16
gttacccata ccaacgtagt atggaggtgg gtacaacaat tcgaccttac agatgtacaa 60
acc                                                               63

<210> 17
<211> 63
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetically generated PCR primers

<400> 17
tactacgttg gtatgggtaa cggtactcaa atttacgtta ttgaccctga accatgtcct 60
gac                                                               63

<210> 18
<211> 57
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetically generated PCR primers

<400> 18
ccggaattct tagtggtggt ggtggtggtg gtcagagtca ggacatggtt cagggtc    57

<210> 19
<211> 45
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetically generated PCR primers

<400> 19
acttctactg agaacaaggt tgctttgact attcaaggtt tgaga                 45

<210> 20
<211> 45
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetically generated PCR primers

<400> 20
tctcaaacct tgaatagtca aagcaacctt gttctcagta gaagt                 45

<210> 21
<211> 45
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetically generated PCR primers

<400> 21
ccatactacg ttggtatggg tgctggtact caaatttacg ttatt                 45

<210> 22
<211> 45
<212> DNA
<213> Artificial Sequence

<220> 
<223> synthetically generated PCR primers

<400> 22
aataacgtaa atttgagtac cagcacccat accaacgtag tatgg                 45

<210> 23
<211> 390
<212> DNA
<213> Artificial Sequence

<220> 
<223> codon optimized pCTLA-4

<400> 23
atgcacgttg ctcaaccagc tgttgtcttg gctaactcta gaggtgttgc ttctttcgtt 60
tgtgagtacg gttctgctgg taaggctgct gaggttagag ttactgtttt gagaagagct 120
ggttctcaaa tgactgaggt ttgtgctgct acttacactg ttgaggacga gttgactttc 180
ttggacgact ctacttgtac tggtacttct actgagaaca aggttaactt gactattcaa 240
ggtttgagag ctgtcgacac cggtttgtac atctgtaagg tcgaattgtt gtacccacct 300
ccatactacg ttggtatggg taacggtact caaatttacg ttattgaccc tgaaccatgt 360
cctgactctg accaccacca ccaccaccac                                  390

<210> 24
<211> 130
<212> PRT
<213> Artificial Sequence

<220> 
<223> porcine CTLA-4

<400> 24
Met His Val Ala Gln Pro Ala Val Val Leu Ala Asn Ser Arg Gly Val
 1               5                  10                  15      
Ala Ser Phe Val Cys Glu Tyr Gly Ser Ala Gly Lys Ala Ala Glu Val
            20                  25                  30          
Arg Val Thr Val Leu Arg Arg Ala Gly Ser Gln Met Thr Glu Val Cys
        35                  40                  45              
Ala Ala Thr Tyr Thr Val Glu Asp Glu Leu Thr Phe Leu Asp Asp Ser
    50                  55                  60                  
Thr Cys Thr Gly Thr Ser Thr Glu Asn Lys Val Asn Leu Thr Ile Gln
65                  70                  75                  80  
Gly Leu Arg Ala Val Asp Thr Gly Leu Tyr Ile Cys Lys Val Glu Leu
                85                  90                  95      
Leu Tyr Pro Pro Pro Tyr Tyr Val Gly Met Gly Asn Gly Thr Gln Ile
            100                 105                 110         
Tyr Val Ile Asp Pro Glu Pro Cys Pro Asp Ser Asp His His His His
        115                 120                 125             
His His
    130 


<210> 25
<211> 390
<212> DNA
<213> Artificial Sequence

<220> 
<223> codon optimized mutant pCTLA-4

<400> 25
atgcacgttg ctcaaccagc tgttgtcttg gctaactcta gaggtgttgc ttctttcgtt 60
tgtgagtacg gttctgctgg taaggctgct gaggttagag ttactgtttt gagaagagct 120
ggttctcaaa tgactgaggt ttgtgctgct acttacactg ttgaggacga gttgactttc 180
ttggacgact ctacttgtac tggtacttct actgagaaca aggttgcttt gactattcaa 240
ggtttgagag ctgtcgacac cggtttgtac atctgtaagg tcgaattgtt gtacccacct 300
ccatactacg ttggtatggg tgctggtact caaatttacg ttattgaccc tgaaccatgt 360
cctgactctg accaccacca ccaccaccac                                  390

<210> 26
<211> 130
<212> PRT
<213> Artificial Sequence

<220> 
<223> mutant pCTLA4

<400> 26
Met His Val Ala Gln Pro Ala Val Val Leu Ala Asn Ser Arg Gly Val
 1               5                  10                  15      
Ala Ser Phe Val Cys Glu Tyr Gly Ser Ala Gly Lys Ala Ala Glu Val
            20                  25                  30          
Arg Val Thr Val Leu Arg Arg Ala Gly Ser Gln Met Thr Glu Val Cys
        35                  40                  45              
Ala Ala Thr Tyr Thr Val Glu Asp Glu Leu Thr Phe Leu Asp Asp Ser
    50                  55                  60                  
Thr Cys Thr Gly Thr Ser Thr Glu Asn Lys Val Ala Leu Thr Ile Gln
65                  70                  75                  80  
Gly Leu Arg Ala Val Asp Thr Gly Leu Tyr Ile Cys Lys Val Glu Leu
                85                  90                  95      
Leu Tyr Pro Pro Pro Tyr Tyr Val Gly Met Gly Ala Gly Thr Gln Ile
            100                 105                 110         
Tyr Val Ile Asp Pro Glu Pro Cys Pro Asp Ser Asp His His His His
        115                 120                 125             
His His
    130 


<210> 27
<211> 1584
<212> DNA
<213> Artificial Sequence

<220> 
<223> DT390-pCTLA-4-Gly-6xHis

<400> 27
gctggtgctg acgacgtcgt cgactcctcc aagtccttcg tcatggagaa cttcgcttcc 60
taccacggga ccaagccagg ttacgtcgac tccatccaga agggtatcca gaagccaaag 120
tccggcaccc aaggtaacta cgacgacgac tggaaggggt tctactccac cgacaacaag 180
tacgacgctg cgggatactc tgtagataat gaaaacccgc tctctggaaa agctggaggc 240
gtggtcaagg tcacctaccc aggtctgact aaggtcttgg ctttgaaggt cgacaacgct 300
gagaccatca agaaggagtt gggtttgtcc ttgactgagc cattgatgga gcaagtcggt 360
accgaagagt tcatcaagag attcggtgac ggtgcttcca gagtcgtctt gtccttgcca 420
ttcgctgagg gttcttctag cgttgaatat attaataact gggaacaggc taaggctttg 480
tctgttgaat tggagattaa cttcgaaacc agaggtaaga gaggtcaaga tgcgatgtat 540
gagtatatgg ctcaagcctg tgctggtaac agagtcagac gttctgttgg ttcctctttg 600
tcctgtatca acctagactg ggacgtcatc agagacaaga ctaagaccaa gatcgagtct 660
ttgaaagagc atggcccaat caagaacaag atgtccgaat cccccgctaa gaccgtctcc 720
gaggaaaagg ccaagcaata cctagaagag ttccaccaaa ccgccttgga gcatcctgaa 780
ttgtcagaac ttaaaaccgt tactgggacc aatcctgtat tcgctggggc taactatgcg 840
gcgtgggcag taaacgttgc gcaagttatc gatagcgaaa cagctgataa tttggaaaag 900
acaactgctg ctctttcgat acttcctggt atcggtagcg taatgggcat tgcagacggt 960
gccgttcacc acaatacaga agagatagtg gcacaatcca tcgctttgtc ctctttgatg 1020
gttgctcaag ctatcccatt ggtcggtgag ttggttgaca tcggtttcgc tgcctacaac 1080
ttcgtcgagt ccatcatcaa cttgttccaa gtcgtccaca actcctacaa ccgtccggct 1140
tactccccag gtcacaagac ccaaccattc ttgccatggg gtggtggtgg ttctatgcac 1200
gttgctcaac cagctgttgt cttggctaac tctagaggtg ttgcttcttt cgtttgtgag 1260
tacggttctg ctggtaaggc tgctgaggtt agagttactg ttttgagaag agctggttct 1320
caaatgactg aggtttgtgc tgctacttac actgttgagg acgagttgac tttcttggac 1380
gactctactt gtactggtac ttctactgag aacaaggtta acttgactat tcaaggtttg 1440
agagctgtcg acaccggttt gtacatctgt aaggtcgaat tgttgtaccc acctccatac 1500
tacgttggta tgggtaacgg tactcaaatt tacgttattg accctgaacc atgtcctgac 1560
tctgaccacc accaccacca ccac                                        1584

<210> 28
<211> 1578
<212> DNA
<213> Artificial Sequence

<220> 
<223> DT390-pCTLA4-Non-N-Gly-6xHis

<400> 28
gctggtgctg acgacgtcgt cgactcctcc aagtccttcg tcatggagaa cttcgcttcc 60
taccacggga ccaagccagg ttacgtcgac tccatccaga agggtatcca gaagccaaag 120
tccggcaccc aaggtaacta cgacgacgac tggaaggggt tctactccac cgacaacaag 180
tacgacgctg cgggatactc tgtagataat gaaaacccgc tctctggaaa agctggaggc 240
gtggtcaagg tcacctaccc aggtctgact aaggtcttgg ctttgaaggt cgacaacgct 300
gagaccatca agaaggagtt gggtttgtcc ttgactgagc cattgatgga gcaagtcggt 360
accgaagagt tcatcaagag attcggtgac ggtgcttcca gagtcgtctt gtccttgcca 420
ttcgctgagg gttcttctag cgttgaatat attaataact gggaacaggc taaggctttg 480
tctgttgaat tggagattaa cttcgaaacc agaggtaaga gaggtcaaga tgcgatgtat 540
gagtatatgg ctcaagcctg tgctggtaac agagtcagac gttctgttgg ttcctctttg 600
tcctgtatca acctagactg ggacgtcatc agagacaaga ctaagaccaa gatcgagtct 660
ttgaaagagc atggcccaat caagaacaag atgtccgaat cccccgctaa gaccgtctcc 720
gaggaaaagg ccaagcaata cctagaagag ttccaccaaa ccgccttgga gcatcctgaa 780
ttgtcagaac ttaaaaccgt tactgggacc aatcctgtat tcgctggggc taactatgcg 840
gcgtgggcag taaacgttgc gcaagttatc gatagcgaaa cagctgataa tttggaaaag 900
acaactgctg ctctttcgat acttcctggt atcggtagcg taatgggcat tgcagacggt 960
gccgttcacc acaatacaga agagatagtg gcacaatcca tcgctttgtc ctctttgatg 1020
gttgctcaag ctatcccatt ggtcggtgag ttggttgaca tcggtttcgc tgcctacaac 1080
ttcgtcgagt ccatcatcaa cttgttccaa gtcgtccaca actcctacaa ccgtccggct 1140
tactccccag gtcacaagac ccaaccattc ttgggtggtg gtggttctat gcacgttgct 1200
caaccagctg ttgtcttggc taactctaga ggtgttgctt ctttcgtttg tgagtacggt 1260
tctgctggta aggctgctga ggttagagtt actgttttga gaagagctgg ttctcaaatg 1320
actgaggttt gtgctgctac ttacactgtt gaggacgagt tgactttctt ggacgactct 1380
acttgtactg gtacttctac tgagaacaag gttgctttga ctattcaagg tttgagagct 1440
gtcgacaccg gtttgtacat ctgtaaggtc gaattgttgt acccacctcc atactacgtt 1500
ggtatgggtg ctggtactca aatttacgtt attgaccctg aaccatgtcc tgactctgac 1560
caccaccacc accaccac                                               1578

<210> 29
<211> 2001
<212> DNA
<213> Artificial Sequence

<220> 
<223> DT390-bi-pCTLA-4-Non-N-Gly-6xHis

<400> 29
gctggtgctg acgacgtcgt cgactcctcc aagtccttcg tcatggagaa cttcgcttcc 60
taccacggga ccaagccagg ttacgtcgac tccatccaga agggtatcca gaagccaaag 120
tccggcaccc aaggtaacta cgacgacgac tggaaggggt tctactccac cgacaacaag 180
tacgacgctg cgggatactc tgtagataat gaaaacccgc tctctggaaa agctggaggc 240
gtggtcaagg tcacctaccc aggtctgact aaggtcttgg ctttgaaggt cgacaacgct 300
gagaccatca agaaggagtt gggtttgtcc ttgactgagc cattgatgga gcaagtcggt 360
accgaagagt tcatcaagag attcggtgac ggtgcttcca gagtcgtctt gtccttgcca 420
ttcgctgagg gttcttctag cgttgaatat attaataact gggaacaggc taaggctttg 480
tctgttgaat tggagattaa cttcgaaacc agaggtaaga gaggtcaaga tgcgatgtat 540
gagtatatgg ctcaagcctg tgctggtaac agagtcagac gttctgttgg ttcctctttg 600
tcctgtatca acctagactg ggacgtcatc agagacaaga ctaagaccaa gatcgagtct 660
ttgaaagagc atggcccaat caagaacaag atgtccgaat cccccgctaa gaccgtctcc 720
gaggaaaagg ccaagcaata cctagaagag ttccaccaaa ccgccttgga gcatcctgaa 780
ttgtcagaac ttaaaaccgt tactgggacc aatcctgtat tcgctggggc taactatgcg 840
gcgtgggcag taaacgttgc gcaagttatc gatagcgaaa cagctgataa tttggaaaag 900
acaactgctg ctctttcgat acttcctggt atcggtagcg taatgggcat tgcagacggt 960
gccgttcacc acaatacaga agagatagtg gcacaatcca tcgctttgtc ctctttgatg 1020
gttgctcaag ctatcccatt ggtcggtgag ttggttgaca tcggtttcgc tgcctacaac 1080
ttcgtcgagt ccatcatcaa cttgttccaa gtcgtccaca actcctacaa ccgtccggct 1140
tactccccag gtcacaagac ccaaccattc ttgccatggg gtggtggtgg ttctatgcac 1200
gttgctcaac cagctgttgt cttggctaac tctagaggtg ttgcttcttt cgtttgtgag 1260
tacggttctg ctggtaaggc tgctgaggtt agagttactg ttttgagaag agctggttct 1320
caaatgactg aggtttgtgc tgctacttac actgttgagg acgagttgac tttcttggac 1380
gactctactt gtactggtac ttctactgag aacaaggttg ctttgactat tcaaggtttg 1440
agagctgtcg acaccggttt gtacatctgt aaggtcgaat tgttgtaccc acctccatac 1500
tacgttggta tgggtgctgg tactcaaatt tacgttattg accctgaacc atgtcctgac 1560
tctgacggtg gtggtggttc tggtggtggt ggatccggtg gtggtggttc tatgcacgtt 1620
gctcaaccag ctgttgtctt ggctaactct agaggtgttg cttctttcgt ttgtgagtac 1680
ggttctgctg gtaaggctgc tgaggttaga gttactgttt tgagaagagc tggttctcaa 1740
atgactgagg tttgtgctgc tacttacact gttgaggacg agttgacttt cttggacgac 1800
tctacttgta ctggtacttc tactgagaac aaggttgctt tgactattca aggtttgaga 1860
gctgtcgaca ccggtttgta catctgtaag gtcgaattgt tgtacccacc tccatactac 1920
gttggtatgg gtgctggtac tcaaatttac gttattgacc ctgaaccatg tcctgactct 1980
gaccaccacc accaccacca c                                           2001

<210> 30
<211> 396
<212> DNA
<213> Artificial Sequence

<220> 
<223> codon optimized Non-gly-Human CTLA-4-6His

<400> 30
aaggctatgc acgttgctca accagctgtt gtcttggctt cttccagagg tattgcttct 60
ttcgtttgtg agtacgcttc tccaggtaag gctactgagg ttagagttac tgtcttgaga 120
caagctgact ctcaagttac tgaggtttgt gctgctactt acatgatggg taacgagttg 180
actttcttgg acgactctat ttgtactggt acttcttccg gtaaccaagt tgctttgact 240
attcaaggtt tgagagctat ggacactggt ttgtacattt gtaaggttga gttgatgtac 300
ccacctccat actacttggg tattggtgct ggtactcaaa tttacgttat tgacccagag 360
ccttgtccag actctgacca ccaccaccac caccac                           396

<210> 31
<211> 132
<212> PRT
<213> Artificial Sequence

<220> 
<223> Non-gly-Human CTLA-4-6His

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

