                         SEQUENCE LISTING

<110>  Seattle Children's Hospital d/b/a Seattle Children's 
       Research Institute
       Jensen, Michael
 
<120>  Bispecific Chimeric Antigen Receptors and Therapeutic Uses 
       Thereof

<130>  067505-000001WO00

<150>  US 61/598,216
<151>  2012-02-13

<160>  15    

<170>  PatentIn version 3.5

<210>  1
<211>  3273
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  GMCSFRss-CD19scFv-Gly4Ser1linker-CD20scFv-IgG4Hinge-CD28tm-41BB-C
       D3zeta-T2A-EGFRt_epHIV7

<400>  1
atgctgctgc tggtgaccag cctgctgctg tgcgagctgc cccaccccgc ctttctgctg       60

atccccatga cccagaccac ctccagcctg agcgccagcc tgggcgaccg ggtgaccatc      120

agctgccggg ccagccagga catcagcaag tacctgaact ggtatcagca gaagcccgac      180

ggcaccgtca agctgctgat ctaccacacc agccggctgc acagcggcgt gcccagccgg      240

tttagcggca gcggctccgg caccgactac agcctgacca tctccaacct ggaacaggaa      300

gatatcgcca cctacttttg ccagcagggc aacacactgc cctacacctt tggcggcgga      360

acaaagctgg aaatcaccgg cagcacctcc ggcagcggca agcctggcag cggcgagggc      420

agcaccaagg gcgaggtgaa gctgcaggaa agcggccctg gcctggtggc ccccagccag      480

agcctgagcg tgacctgcac cgtgagcggc gtgagcctgc ccgactacgg cgtgagctgg      540

atccggcagc cccccaggaa gggcctggaa tggctgggcg tgatctgggg cagcgagacc      600

acctactaca acagcgccct gaagagccgg ctgaccatca tcaaggacaa cagcaagagc      660

caggtgttcc tgaagatgaa cagcctgcag accgacgaca ccgccatcta ctactgcgcc      720

aagcactact actacggcgg cagctacgcc atggactact ggggccaggg caccagcgtg      780

accgtgagca gcggaggtgg tggatccgag gtgcagctgc agcagtctgg ggctgagctg      840

gtgaagcctg gggcctcagt gaagatgtcc tgcaaggctt ctggctacac atttaccagt      900

tacaatatgc actgggtaaa gcagacacct ggacagggcc tggaatggat tggagctatt      960

tatccaggaa atggtgatac ttcctacaat cagaagttca aaggcaaggc cacattgact     1020

gcagacaaat cctccagcac agcctacatg cagctcagca gcctgacatc tgaggactct     1080

gcggactatt actgtgcaag atctaattat tacggtagta gctactggtt cttcgatgtc     1140

tggggcgcag ggaccacggt caccgtctcc tcaggcagta ctagcggtgg tggctccggg     1200

ggcggttccg gtgggggcgg cagcagcgac attgtgctga cccaatctcc agctatcctg     1260

tctgcatctc caggggagaa ggtcacaatg acttgcaggg ccagctcaag tgtaaattac     1320

atggactggt accagaagaa gccaggatcc tcccccaaac cctggattta tgccacatcc     1380

aacctggctt ctggagtccc tgctcgcttc agtggcagtg ggtctgggac ctcttactct     1440

ctcacaatca gcagagtgga ggctgaagat gctgccactt attactgcca gcagtggagt     1500

tttaatccac ccacgttcgg aggggggacc aagctggaaa taaaagagag caagtacgga     1560

ccgccctgcc ccccttgccc tatgttctgg gtgctggtgg tggtcggagg cgtgctggcc     1620

tgctacagcc tgctggtcac cgtggccttc atcatctttt gggtgaaacg gggcagaaag     1680

aaactcctgt atatattcaa acaaccattt atgagaccag tacaaactac tcaagaggaa     1740

gatggctgta gctgccgatt tccagaagaa gaagaaggag gatgtgaact gcgggtgaag     1800

ttcagcagaa gcgccgacgc ccctgcctac cagcagggcc agaatcagct gtacaacgag     1860

ctgaacctgg gcagaaggga agagtacgac gtcctggata agcggagagg ccgggaccct     1920

gagatgggcg gcaagcctcg gcggaagaac ccccaggaag gcctgtataa cgaactgcag     1980

aaagacaaga tggccgaggc ctacagcgag atcggcatga agggcgagcg gaggcggggc     2040

aagggccacg acggcctgta tcagggcctg tccaccgcca ccaaggatac ctacgacgcc     2100

ctgcacatgc aggccctgcc cccaaggctc gagggcggcg gagagggcag aggaagtctt     2160

ctaacatgcg gtgacgtgga ggagaatccc ggccctagga tgcttctcct ggtgacaagc     2220

cttctgctct gtgagttacc acacccagca ttcctcctga tcccacgcaa agtgtgtaac     2280

ggaataggta ttggtgaatt taaagactca ctctccataa atgctacgaa tattaaacac     2340

ttcaaaaact gcacctccat cagtggcgat ctccacatcc tgccggtggc atttaggggt     2400

gactccttca cacatactcc tcctctggat ccacaggaac tggatattct gaaaaccgta     2460

aaggaaatca cagggttttt gctgattcag gcttggcctg aaaacaggac ggacctccat     2520

gcctttgaga acctagaaat catacgcggc aggaccaagc aacatggtca gttttctctt     2580

gcagtcgtca gcctgaacat aacatccttg ggattacgct ccctcaagga gataagtgat     2640

ggagatgtga taatttcagg aaacaaaaat ttgtgctatg caaatacaat aaactggaaa     2700

aaactgtttg ggacctccgg tcagaaaacc aaaattataa gcaacagagg tgaaaacagc     2760

tgcaaggcca caggccaggt ctgccatgcc ttgtgctccc ccgagggctg ctggggcccg     2820

gagcccaggg actgcgtctc ttgccggaat gtcagccgag gcagggaatg cgtggacaag     2880

tgcaaccttc tggagggtga gccaagggag tttgtggaga actctgagtg catacagtgc     2940

cacccagagt gcctgcctca ggccatgaac atcacctgca caggacgggg accagacaac     3000

tgtatccagt gtgcccacta cattgacggc ccccactgcg tcaagacctg cccggcagga     3060

gtcatgggag aaaacaacac cctggtctgg aagtacgcag acgccggcca tgtgtgccac     3120

ctgtgccatc caaactgcac ctacggatgc actgggccag gtcttgaagg ctgtccaacg     3180

aatgggccta agatcccgtc catcgccact gggatggtgg gggccctcct cttgctgctg     3240

gtggtggccc tggggatcgg cctcttcatg tga                                  3273


<210>  2
<211>  3273
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  GMCSFRss-CD19scFv-Gly4Ser1linker-CD20scFv-IgG4Hinge-CD28tm-41BB-C
       D3zeta-T2A-EGFRt_epHIV7


<220>
<221>  CDS
<222>  (1)..(3273)

<400>  2
atg ctg ctg ctg gtg acc agc ctg ctg ctg tgc gag ctg ccc cac ccc         48
Met Leu Leu Leu Val Thr Ser Leu Leu Leu Cys Glu Leu Pro His Pro           
1               5                   10                  15                

gcc ttt ctg ctg atc ccc atg acc cag acc acc tcc agc ctg agc gcc         96
Ala Phe Leu Leu Ile Pro Met Thr Gln Thr Thr Ser Ser Leu Ser Ala           
            20                  25                  30                    

agc ctg ggc gac cgg gtg acc atc agc tgc cgg gcc agc cag gac atc        144
Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Asp Ile           
        35                  40                  45                        

agc aag tac ctg aac tgg tat cag cag aag ccc gac ggc acc gtc aag        192
Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys           
    50                  55                  60                            

ctg ctg atc tac cac acc agc cgg ctg cac agc ggc gtg ccc agc cgg        240
Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg           
65                  70                  75                  80            

ttt agc ggc agc ggc tcc ggc acc gac tac agc ctg acc atc tcc aac        288
Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn           
                85                  90                  95                

ctg gaa cag gaa gat atc gcc acc tac ttt tgc cag cag ggc aac aca        336
Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly Asn Thr           
            100                 105                 110                   

ctg ccc tac acc ttt ggc ggc gga aca aag ctg gaa atc acc ggc agc        384
Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Thr Gly Ser           
        115                 120                 125                       

acc tcc ggc agc ggc aag cct ggc agc ggc gag ggc agc acc aag ggc        432
Thr Ser Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys Gly           
    130                 135                 140                           

gag gtg aag ctg cag gaa agc ggc cct ggc ctg gtg gcc ccc agc cag        480
Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln           
145                 150                 155                 160           

agc ctg agc gtg acc tgc acc gtg agc ggc gtg agc ctg ccc gac tac        528
Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr           
                165                 170                 175               

ggc gtg agc tgg atc cgg cag ccc ccc agg aag ggc ctg gaa tgg ctg        576
Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu           
            180                 185                 190                   

ggc gtg atc tgg ggc agc gag acc acc tac tac aac agc gcc ctg aag        624
Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser Ala Leu Lys           
        195                 200                 205                       

agc cgg ctg acc atc atc aag gac aac agc aag agc cag gtg ttc ctg        672
Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu           
    210                 215                 220                           

aag atg aac agc ctg cag acc gac gac acc gcc atc tac tac tgc gcc        720
Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala           
225                 230                 235                 240           

aag cac tac tac tac ggc ggc agc tac gcc atg gac tac tgg ggc cag        768
Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln           
                245                 250                 255               

ggc acc agc gtg acc gtg agc agc gga ggt ggt gga tcc gag gtg cag        816
Gly Thr Ser Val Thr Val Ser Ser Gly Gly Gly Gly Ser Glu Val Gln           
            260                 265                 270                   

ctg cag cag tct ggg gct gag ctg gtg aag cct ggg gcc tca gtg aag        864
Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala Ser Val Lys           
        275                 280                 285                       

atg tcc tgc aag gct tct ggc tac aca ttt acc agt tac aat atg cac        912
Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr Asn Met His           
    290                 295                 300                           

tgg gta aag cag aca cct gga cag ggc ctg gaa tgg att gga gct att        960
Trp Val Lys Gln Thr Pro Gly Gln Gly Leu Glu Trp Ile Gly Ala Ile           
305                 310                 315                 320           

tat cca gga aat ggt gat act tcc tac aat cag aag ttc aaa ggc aag       1008
Tyr Pro Gly Asn Gly Asp Thr Ser Tyr Asn Gln Lys Phe Lys Gly Lys           
                325                 330                 335               

gcc aca ttg act gca gac aaa tcc tcc agc aca gcc tac atg cag ctc       1056
Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr Met Gln Leu           
            340                 345                 350                   

agc agc ctg aca tct gag gac tct gcg gac tat tac tgt gca aga tct       1104
Ser Ser Leu Thr Ser Glu Asp Ser Ala Asp Tyr Tyr Cys Ala Arg Ser           
        355                 360                 365                       

aat tat tac ggt agt agc tac tgg ttc ttc gat gtc tgg ggc gca ggg       1152
Asn Tyr Tyr Gly Ser Ser Tyr Trp Phe Phe Asp Val Trp Gly Ala Gly           
    370                 375                 380                           

acc acg gtc acc gtc tcc tca ggc agt act agc ggt ggt ggc tcc ggg       1200
Thr Thr Val Thr Val Ser Ser Gly Ser Thr Ser Gly Gly Gly Ser Gly           
385                 390                 395                 400           

ggc ggt tcc ggt ggg ggc ggc agc agc gac att gtg ctg acc caa tct       1248
Gly Gly Ser Gly Gly Gly Gly Ser Ser Asp Ile Val Leu Thr Gln Ser           
                405                 410                 415               

cca gct atc ctg tct gca tct cca ggg gag aag gtc aca atg act tgc       1296
Pro Ala Ile Leu Ser Ala Ser Pro Gly Glu Lys Val Thr Met Thr Cys           
            420                 425                 430                   

agg gcc agc tca agt gta aat tac atg gac tgg tac cag aag aag cca       1344
Arg Ala Ser Ser Ser Val Asn Tyr Met Asp Trp Tyr Gln Lys Lys Pro           
        435                 440                 445                       

gga tcc tcc ccc aaa ccc tgg att tat gcc aca tcc aac ctg gct tct       1392
Gly Ser Ser Pro Lys Pro Trp Ile Tyr Ala Thr Ser Asn Leu Ala Ser           
    450                 455                 460                           

gga gtc cct gct cgc ttc agt ggc agt ggg tct ggg acc tct tac tct       1440
Gly Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Ser Tyr Ser           
465                 470                 475                 480           

ctc aca atc agc aga gtg gag gct gaa gat gct gcc act tat tac tgc       1488
Leu Thr Ile Ser Arg Val Glu Ala Glu Asp Ala Ala Thr Tyr Tyr Cys           
                485                 490                 495               

cag cag tgg agt ttt aat cca ccc acg ttc gga ggg ggg acc aag ctg       1536
Gln Gln Trp Ser Phe Asn Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu           
            500                 505                 510                   

gaa ata aaa gag agc aag tac gga ccg ccc tgc ccc cct tgc cct atg       1584
Glu Ile Lys Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Met           
        515                 520                 525                       

ttc tgg gtg ctg gtg gtg gtc gga ggc gtg ctg gcc tgc tac agc ctg       1632
Phe Trp Val Leu Val Val Val Gly Gly Val Leu Ala Cys Tyr Ser Leu           
    530                 535                 540                           

ctg gtc acc gtg gcc ttc atc atc ttt tgg gtg aaa cgg ggc aga aag       1680
Leu Val Thr Val Ala Phe Ile Ile Phe Trp Val Lys Arg Gly Arg Lys           
545                 550                 555                 560           

aaa ctc ctg tat ata ttc aaa caa cca ttt atg aga cca gta caa act       1728
Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val Gln Thr           
                565                 570                 575               

act caa gag gaa gat ggc tgt agc tgc cga ttt cca gaa gaa gaa gaa       1776
Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu           
            580                 585                 590                   

gga gga tgt gaa ctg cgg gtg aag ttc agc aga agc gcc gac gcc cct       1824
Gly Gly Cys Glu Leu Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro           
        595                 600                 605                       

gcc tac cag cag ggc cag aat cag ctg tac aac gag ctg aac ctg ggc       1872
Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly           
    610                 615                 620                           

aga agg gaa gag tac gac gtc ctg gat aag cgg aga ggc cgg gac cct       1920
Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro           
625                 630                 635                 640           

gag atg ggc ggc aag cct cgg cgg aag aac ccc cag gaa ggc ctg tat       1968
Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr           
                645                 650                 655               

aac gaa ctg cag aaa gac aag atg gcc gag gcc tac agc gag atc ggc       2016
Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly           
            660                 665                 670                   

atg aag ggc gag cgg agg cgg ggc aag ggc cac gac ggc ctg tat cag       2064
Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln           
        675                 680                 685                       

ggc ctg tcc acc gcc acc aag gat acc tac gac gcc ctg cac atg cag       2112
Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln           
    690                 695                 700                           

gcc ctg ccc cca agg ctc gag ggc ggc gga gag ggc aga gga agt ctt       2160
Ala Leu Pro Pro Arg Leu Glu Gly Gly Gly Glu Gly Arg Gly Ser Leu           
705                 710                 715                 720           

cta aca tgc ggt gac gtg gag gag aat ccc ggc cct agg atg ctt ctc       2208
Leu Thr Cys Gly Asp Val Glu Glu Asn Pro Gly Pro Arg Met Leu Leu           
                725                 730                 735               

ctg gtg aca agc ctt ctg ctc tgt gag tta cca cac cca gca ttc ctc       2256
Leu Val Thr Ser Leu Leu Leu Cys Glu Leu Pro His Pro Ala Phe Leu           
            740                 745                 750                   

ctg atc cca cgc aaa gtg tgt aac gga ata ggt att ggt gaa ttt aaa       2304
Leu Ile Pro Arg Lys Val Cys Asn Gly Ile Gly Ile Gly Glu Phe Lys           
        755                 760                 765                       

gac tca ctc tcc ata aat gct acg aat att aaa cac ttc aaa aac tgc       2352
Asp Ser Leu Ser Ile Asn Ala Thr Asn Ile Lys His Phe Lys Asn Cys           
    770                 775                 780                           

acc tcc atc agt ggc gat ctc cac atc ctg ccg gtg gca ttt agg ggt       2400
Thr Ser Ile Ser Gly Asp Leu His Ile Leu Pro Val Ala Phe Arg Gly           
785                 790                 795                 800           

gac tcc ttc aca cat act cct cct ctg gat cca cag gaa ctg gat att       2448
Asp Ser Phe Thr His Thr Pro Pro Leu Asp Pro Gln Glu Leu Asp Ile           
                805                 810                 815               

ctg aaa acc gta aag gaa atc aca ggg ttt ttg ctg att cag gct tgg       2496
Leu Lys Thr Val Lys Glu Ile Thr Gly Phe Leu Leu Ile Gln Ala Trp           
            820                 825                 830                   

cct gaa aac agg acg gac ctc cat gcc ttt gag aac cta gaa atc ata       2544
Pro Glu Asn Arg Thr Asp Leu His Ala Phe Glu Asn Leu Glu Ile Ile           
        835                 840                 845                       

cgc ggc agg acc aag caa cat ggt cag ttt tct ctt gca gtc gtc agc       2592
Arg Gly Arg Thr Lys Gln His Gly Gln Phe Ser Leu Ala Val Val Ser           
    850                 855                 860                           

ctg aac ata aca tcc ttg gga tta cgc tcc ctc aag gag ata agt gat       2640
Leu Asn Ile Thr Ser Leu Gly Leu Arg Ser Leu Lys Glu Ile Ser Asp           
865                 870                 875                 880           

gga gat gtg ata att tca gga aac aaa aat ttg tgc tat gca aat aca       2688
Gly Asp Val Ile Ile Ser Gly Asn Lys Asn Leu Cys Tyr Ala Asn Thr           
                885                 890                 895               

ata aac tgg aaa aaa ctg ttt ggg acc tcc ggt cag aaa acc aaa att       2736
Ile Asn Trp Lys Lys Leu Phe Gly Thr Ser Gly Gln Lys Thr Lys Ile           
            900                 905                 910                   

ata agc aac aga ggt gaa aac agc tgc aag gcc aca ggc cag gtc tgc       2784
Ile Ser Asn Arg Gly Glu Asn Ser Cys Lys Ala Thr Gly Gln Val Cys           
        915                 920                 925                       

cat gcc ttg tgc tcc ccc gag ggc tgc tgg ggc ccg gag ccc agg gac       2832
His Ala Leu Cys Ser Pro Glu Gly Cys Trp Gly Pro Glu Pro Arg Asp           
    930                 935                 940                           

tgc gtc tct tgc cgg aat gtc agc cga ggc agg gaa tgc gtg gac aag       2880
Cys Val Ser Cys Arg Asn Val Ser Arg Gly Arg Glu Cys Val Asp Lys           
945                 950                 955                 960           

tgc aac ctt ctg gag ggt gag cca agg gag ttt gtg gag aac tct gag       2928
Cys Asn Leu Leu Glu Gly Glu Pro Arg Glu Phe Val Glu Asn Ser Glu           
                965                 970                 975               

tgc ata cag tgc cac cca gag tgc ctg cct cag gcc atg aac atc acc       2976
Cys Ile Gln Cys His Pro Glu Cys Leu Pro Gln Ala Met Asn Ile Thr           
            980                 985                 990                   

tgc aca gga cgg gga cca gac aac  tgt atc cag tgt gcc  cac tac att     3024
Cys Thr Gly Arg Gly Pro Asp Asn  Cys Ile Gln Cys Ala  His Tyr Ile         
        995                 1000                 1005                     

gac ggc  ccc cac tgc gtc aag  acc tgc ccg gca gga  gtc atg gga        3069
Asp Gly  Pro His Cys Val Lys  Thr Cys Pro Ala Gly  Val Met Gly            
    1010                 1015                 1020                        

gaa aac  aac acc ctg gtc tgg  aag tac gca gac gcc  ggc cat gtg        3114
Glu Asn  Asn Thr Leu Val Trp  Lys Tyr Ala Asp Ala  Gly His Val            
    1025                 1030                 1035                        

tgc cac  ctg tgc cat cca aac  tgc acc tac gga tgc  act ggg cca        3159
Cys His  Leu Cys His Pro Asn  Cys Thr Tyr Gly Cys  Thr Gly Pro            
    1040                 1045                 1050                        

ggt ctt  gaa ggc tgt cca acg  aat ggg cct aag atc  ccg tcc atc        3204
Gly Leu  Glu Gly Cys Pro Thr  Asn Gly Pro Lys Ile  Pro Ser Ile            
    1055                 1060                 1065                        

gcc act  ggg atg gtg ggg gcc  ctc ctc ttg ctg ctg  gtg gtg gcc        3249
Ala Thr  Gly Met Val Gly Ala  Leu Leu Leu Leu Leu  Val Val Ala            
    1070                 1075                 1080                        

ctg ggg  atc ggc ctc ttc atg  tga                                     3273
Leu Gly  Ile Gly Leu Phe Met                                              
    1085                 1090                                             


<210>  3
<211>  1090
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic Construct

<400>  3

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 Met Thr Gln Thr Thr Ser Ser Leu Ser Ala 
            20                  25                  30          


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


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


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


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


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


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


Thr Ser Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser Thr Lys Gly 
    130                 135                 140                 


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


Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu Pro Asp Tyr 
                165                 170                 175     


Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu Glu Trp Leu 
            180                 185                 190         


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


Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln Val Phe Leu 
    210                 215                 220                 


Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala 
225                 230                 235                 240 


Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr Trp Gly Gln 
                245                 250                 255     


Gly Thr Ser Val Thr Val Ser Ser Gly Gly Gly Gly Ser Glu Val Gln 
            260                 265                 270         


Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala Ser Val Lys 
        275                 280                 285             


Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr Asn Met His 
    290                 295                 300                 


Trp Val Lys Gln Thr Pro Gly Gln Gly Leu Glu Trp Ile Gly Ala Ile 
305                 310                 315                 320 


Tyr Pro Gly Asn Gly Asp Thr Ser Tyr Asn Gln Lys Phe Lys Gly Lys 
                325                 330                 335     


Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr Met Gln Leu 
            340                 345                 350         


Ser Ser Leu Thr Ser Glu Asp Ser Ala Asp Tyr Tyr Cys Ala Arg Ser 
        355                 360                 365             


Asn Tyr Tyr Gly Ser Ser Tyr Trp Phe Phe Asp Val Trp Gly Ala Gly 
    370                 375                 380                 


Thr Thr Val Thr Val Ser Ser Gly Ser Thr Ser Gly Gly Gly Ser Gly 
385                 390                 395                 400 


Gly Gly Ser Gly Gly Gly Gly Ser Ser Asp Ile Val Leu Thr Gln Ser 
                405                 410                 415     


Pro Ala Ile Leu Ser Ala Ser Pro Gly Glu Lys Val Thr Met Thr Cys 
            420                 425                 430         


Arg Ala Ser Ser Ser Val Asn Tyr Met Asp Trp Tyr Gln Lys Lys Pro 
        435                 440                 445             


Gly Ser Ser Pro Lys Pro Trp Ile Tyr Ala Thr Ser Asn Leu Ala Ser 
    450                 455                 460                 


Gly Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr Ser Tyr Ser 
465                 470                 475                 480 


Leu Thr Ile Ser Arg Val Glu Ala Glu Asp Ala Ala Thr Tyr Tyr Cys 
                485                 490                 495     


Gln Gln Trp Ser Phe Asn Pro Pro Thr Phe Gly Gly Gly Thr Lys Leu 
            500                 505                 510         


Glu Ile Lys Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Met 
        515                 520                 525             


Phe Trp Val Leu Val Val Val Gly Gly Val Leu Ala Cys Tyr Ser Leu 
    530                 535                 540                 


Leu Val Thr Val Ala Phe Ile Ile Phe Trp Val Lys Arg Gly Arg Lys 
545                 550                 555                 560 


Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val Gln Thr 
                565                 570                 575     


Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu 
            580                 585                 590         


Gly Gly Cys Glu Leu Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro 
        595                 600                 605             


Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly 
    610                 615                 620                 


Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro 
625                 630                 635                 640 


Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr 
                645                 650                 655     


Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly 
            660                 665                 670         


Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln 
        675                 680                 685             


Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln 
    690                 695                 700                 


Ala Leu Pro Pro Arg Leu Glu Gly Gly Gly Glu Gly Arg Gly Ser Leu 
705                 710                 715                 720 


Leu Thr Cys Gly Asp Val Glu Glu Asn Pro Gly Pro Arg Met Leu Leu 
                725                 730                 735     


Leu Val Thr Ser Leu Leu Leu Cys Glu Leu Pro His Pro Ala Phe Leu 
            740                 745                 750         


Leu Ile Pro Arg Lys Val Cys Asn Gly Ile Gly Ile Gly Glu Phe Lys 
        755                 760                 765             


Asp Ser Leu Ser Ile Asn Ala Thr Asn Ile Lys His Phe Lys Asn Cys 
    770                 775                 780                 


Thr Ser Ile Ser Gly Asp Leu His Ile Leu Pro Val Ala Phe Arg Gly 
785                 790                 795                 800 


Asp Ser Phe Thr His Thr Pro Pro Leu Asp Pro Gln Glu Leu Asp Ile 
                805                 810                 815     


Leu Lys Thr Val Lys Glu Ile Thr Gly Phe Leu Leu Ile Gln Ala Trp 
            820                 825                 830         


Pro Glu Asn Arg Thr Asp Leu His Ala Phe Glu Asn Leu Glu Ile Ile 
        835                 840                 845             


Arg Gly Arg Thr Lys Gln His Gly Gln Phe Ser Leu Ala Val Val Ser 
    850                 855                 860                 


Leu Asn Ile Thr Ser Leu Gly Leu Arg Ser Leu Lys Glu Ile Ser Asp 
865                 870                 875                 880 


Gly Asp Val Ile Ile Ser Gly Asn Lys Asn Leu Cys Tyr Ala Asn Thr 
                885                 890                 895     


Ile Asn Trp Lys Lys Leu Phe Gly Thr Ser Gly Gln Lys Thr Lys Ile 
            900                 905                 910         


Ile Ser Asn Arg Gly Glu Asn Ser Cys Lys Ala Thr Gly Gln Val Cys 
        915                 920                 925             


His Ala Leu Cys Ser Pro Glu Gly Cys Trp Gly Pro Glu Pro Arg Asp 
    930                 935                 940                 


Cys Val Ser Cys Arg Asn Val Ser Arg Gly Arg Glu Cys Val Asp Lys 
945                 950                 955                 960 


Cys Asn Leu Leu Glu Gly Glu Pro Arg Glu Phe Val Glu Asn Ser Glu 
                965                 970                 975     


Cys Ile Gln Cys His Pro Glu Cys Leu Pro Gln Ala Met Asn Ile Thr 
            980                 985                 990         


Cys Thr Gly Arg Gly Pro Asp Asn  Cys Ile Gln Cys Ala  His Tyr Ile 
        995                 1000                 1005             


Asp Gly  Pro His Cys Val Lys  Thr Cys Pro Ala Gly  Val Met Gly 
    1010                 1015                 1020             


Glu Asn  Asn Thr Leu Val Trp  Lys Tyr Ala Asp Ala  Gly His Val 
    1025                 1030                 1035             


Cys His  Leu Cys His Pro Asn  Cys Thr Tyr Gly Cys  Thr Gly Pro 
    1040                 1045                 1050             


Gly Leu  Glu Gly Cys Pro Thr  Asn Gly Pro Lys Ile  Pro Ser Ile 
    1055                 1060                 1065             


Ala Thr  Gly Met Val Gly Ala  Leu Leu Leu Leu Leu  Val Val Ala 
    1070                 1075                 1080             


Leu Gly  Ile Gly Leu Phe Met  
    1085                 1090 


<210>  4
<211>  582
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  IgG4hinge-CD28tm-41BB-CD3Zeta

<400>  4
gagagcaagt acggaccgcc ctgcccccct tgccctatgt tctgggtgct ggtggtggtc       60

ggaggcgtgc tggcctgcta cagcctgctg gtcaccgtgg ccttcatcat cttttgggtg      120

aaacggggca gaaagaaact cctgtatata ttcaaacaac catttatgag accagtacaa      180

actactcaag aggaagatgg ctgtagctgc cgatttccag aagaagaaga aggaggatgt      240

gaactgcggg tgaagttcag cagaagcgcc gacgcccctg cctaccagca gggccagaat      300

cagctgtaca acgagctgaa cctgggcaga agggaagagt acgacgtcct ggataagcgg      360

agaggccggg accctgagat gggcggcaag cctcggcgga agaaccccca ggaaggcctg      420

tataacgaac tgcagaaaga caagatggcc gaggcctaca gcgagatcgg catgaagggc      480

gagcggaggc ggggcaaggg ccacgacggc ctgtatcagg gcctgtccac cgccaccaag      540

gatacctacg acgccctgca catgcaggcc ctgcccccaa gg                         582


<210>  5
<211>  582
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  IgG4hinge-CD28tm-41BB-CD3Zeta


<220>
<221>  CDS
<222>  (1)..(582)

<400>  5
gag agc aag tac gga ccg ccc tgc ccc cct tgc cct atg ttc tgg gtg         48
Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Met Phe Trp Val           
1               5                   10                  15                

ctg gtg gtg gtc gga ggc gtg ctg gcc tgc tac agc ctg ctg gtc acc         96
Leu Val Val Val Gly Gly Val Leu Ala Cys Tyr Ser Leu Leu Val Thr           
            20                  25                  30                    

gtg gcc ttc atc atc ttt tgg gtg aaa cgg ggc aga aag aaa ctc ctg        144
Val Ala Phe Ile Ile Phe Trp Val Lys Arg Gly Arg Lys Lys Leu Leu           
        35                  40                  45                        

tat ata ttc aaa caa cca ttt atg aga cca gta caa act act caa gag        192
Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val Gln Thr Thr Gln Glu           
    50                  55                  60                            

gaa gat ggc tgt agc tgc cga ttt cca gaa gaa gaa gaa gga gga tgt        240
Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys           
65                  70                  75                  80            

gaa ctg cgg gtg aag ttc agc aga agc gcc gac gcc cct gcc tac cag        288
Glu Leu Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln           
                85                  90                  95                

cag ggc cag aat cag ctg tac aac gag ctg aac ctg ggc aga agg gaa        336
Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu           
            100                 105                 110                   

gag tac gac gtc ctg gat aag cgg aga ggc cgg gac cct gag atg ggc        384
Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly           
        115                 120                 125                       

ggc aag cct cgg cgg aag aac ccc cag gaa ggc ctg tat aac gaa ctg        432
Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu           
    130                 135                 140                           

cag aaa gac aag atg gcc gag gcc tac agc gag atc ggc atg aag ggc        480
Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly           
145                 150                 155                 160           

gag cgg agg cgg ggc aag ggc cac gac ggc ctg tat cag ggc ctg tcc        528
Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser           
                165                 170                 175               

acc gcc acc aag gat acc tac gac gcc ctg cac atg cag gcc ctg ccc        576
Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro           
            180                 185                 190                   

cca agg                                                                582
Pro Arg                                                                   
                                                                          


<210>  6
<211>  194
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic Construct

<400>  6

Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Met Phe Trp Val 
1               5                   10                  15      


Leu Val Val Val Gly Gly Val Leu Ala Cys Tyr Ser Leu Leu Val Thr 
            20                  25                  30          


Val Ala Phe Ile Ile Phe Trp Val Lys Arg Gly Arg Lys Lys Leu Leu 
        35                  40                  45              


Tyr Ile Phe Lys Gln Pro Phe Met Arg Pro Val Gln Thr Thr Gln Glu 
    50                  55                  60                  


Glu Asp Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys 
65                  70                  75                  80  


Glu Leu Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln 
                85                  90                  95      


Gln Gly Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu 
            100                 105                 110         


Glu Tyr Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly 
        115                 120                 125             


Gly Lys Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu 
    130                 135                 140                 


Gln Lys Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly 
145                 150                 155                 160 


Glu Arg Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser 
                165                 170                 175     


Thr Ala Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro 
            180                 185                 190         


Pro Arg 
        


<210>  7
<211>  2910
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  GMCSFRss-CD19scFv-Gly4serlinker-CD20scFv-huIgG4hinge/CH2/CH3-CD28
       tm/CD28cyto-41BB-CD3Zeta

<400>  7
atgctgctgc tggtgaccag cctgctgctg tgcgagctgc cccaccccgc ctttctgctg       60

atccccgaca tccagatgac ccagaccacc tccagcctga gcgccagcct gggcgaccgg      120

gtgaccatca gctgccgggc cagccaggac atcagcaagt acctgaactg gtatcagcag      180

aagcccgacg gcaccgtcaa gctgctgatc taccacacca gccggctgca cagcggcgtg      240

cccagccggt ttagcggcag cggctccggc accgactaca gcctgaccat ctccaacctg      300

gaacaggaag atatcgccac ctacttttgc cagcagggca acacactgcc ctacaccttt      360

ggcggcggaa caaagctgga aatcaccggc agcacctccg gcagcggcaa gcctggcagc      420

ggcgagggca gcaccaaggg cgaggtgaag ctgcaggaaa gcggccctgg cctggtggcc      480

cccagccaga gcctgagcgt gacctgcacc gtgagcggcg tgagcctgcc cgactacggc      540

gtgagctgga tccggcagcc ccccaggaag ggcctggaat ggctgggcgt gatctggggc      600

agcgagacca cctactacaa cagcgccctg aagagccggc tgaccatcat caaggacaac      660

agcaagagcc aggtgttcct gaagatgaac agcctgcaga ccgacgacac cgccatctac      720

tactgcgcca agcactacta ctacggcggc agctacgcca tggactactg gggccagggc      780

accagcgtga ccgtgagcag cggaggtggt ggatccgagg tgcagctgca gcagtctggg      840

gctgagctgg tgaagcctgg ggcctcagtg aagatgtcct gcaaggcttc tggctacaca      900

tttaccagtt acaatatgca ctgggtaaag cagacacctg gacagggcct ggaatggatt      960

ggagctattt atccaggaaa tggtgatact tcctacaatc agaagttcaa aggcaaggcc     1020

acattgactg cagacaaatc ctccagcaca gcctacatgc agctcagcag cctgacatct     1080

gaggactctg cggactatta ctgtgcaaga tctaattatt acggtagtag ctactggttc     1140

ttcgatgtct ggggcgcagg gaccacggtc accgtctcct caggcagtac tagcggtggt     1200

ggctccgggg gcggttccgg tgggggcggc agcagcgaca ttgtgctgac ccaatctcca     1260

gctatcctgt ctgcatctcc aggggagaag gtcacaatga cttgcagggc cagctcaagt     1320

gtaaattaca tggactggta ccagaagaag ccaggatcct cccccaaacc ctggatttat     1380

gccacatcca acctggcttc tggagtccct gctcgcttca gtggcagtgg gtctgggacc     1440

tcttactctc tcacaatcag cagagtggag gctgaagatg ctgccactta ttactgccag     1500

cagtggagtt ttaatccacc cacgttcgga ggggggacca agctggaaat aaaagagagc     1560

aagtacggac cgccctgccc cccttgccct gcccccgagt tcctgggcgg acccagcgtg     1620

ttcctgttcc cccccaagcc caaggacacc ctgatgatca gccggacccc cgaggtgacc     1680

tgcgtggtgg tggacgtgag ccaggaagat cccgaggtcc agttcaattg gtacgtggac     1740

ggcgtggaag tgcacaacgc caagaccaag cccagagagg aacagttcaa cagcacctac     1800

cgggtggtgt ctgtgctgac cgtgctgcac caggactggc tgaacggcaa agaatacaag     1860

tgcaaggtgt ccaacaaggg cctgcccagc agcatcgaaa agaccatcag caaggccaag     1920

ggccagcctc gcgagcccca ggtgtacacc ctgcctccct cccaggaaga gatgaccaag     1980

aaccaggtgt ccctgacctg cctggtgaag ggcttctacc ccagcgacat cgccgtggag     2040

tgggagagca acggccagcc tgagaacaac tacaagacca cccctcccgt gctggacagc     2100

gacggcagct tcttcctgta cagccggctg accgtggaca agagccggtg gcaggaaggc     2160

aacgtcttta gctgcagcgt gatgcacgag gccctgcaca accactacac ccagaagagc     2220

ctgagcctgt ccctgggcaa gatgttctgg gtgctggtgg tggtgggcgg ggtgctggcc     2280

tgctacagcc tgctggtgac agtggccttc atcatctttt gggtgcggag caagcggagc     2340

agaggcggcc acagcgacta catgaacatg acccccagac ggcctggccc cacccggaag     2400

cactaccagc cctacgcccc acccagggac tttgccgcct acagaagcaa acggggcaga     2460

aagaaactcc tgtatatatt caaacaacca tttatgagac cagtacaaac tactcaagag     2520

gaagatggct gtagctgccg atttccagaa gaagaagaag gaggatgtga actgcgggtg     2580

aagttcagca gaagcgccga cgcccctgcc taccagcagg gccagaatca gctgtacaac     2640

gagctgaacc tgggcagaag ggaagagtac gacgtcctgg ataagcggag aggccgggac     2700

cctgagatgg gcggcaagcc tcggcggaag aacccccagg aaggcctgta taacgaactg     2760

cagaaagaca agatggccga ggcctacagc gagatcggca tgaagggcga gcggaggcgg     2820

ggcaagggcc acgacggcct gtatcagggc ctgtccaccg ccaccaagga tacctacgac     2880

gccctgcaca tgcaggccct gcccccaagg                                      2910


<210>  8
<211>  2910
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  GMCSFR-ssCD19scFv-Gly4serlinker-CD20scFv-huIgG4hinge/CH2/CH3-CD28
       tm/CD28cyto-41BB-CD3Zeta


<220>
<221>  CDS
<222>  (1)..(2910)

<400>  8
atg ctg ctg ctg gtg acc agc ctg ctg ctg tgc gag ctg ccc cac ccc         48
Met Leu Leu Leu Val Thr Ser Leu Leu Leu Cys Glu Leu Pro His Pro           
1               5                   10                  15                

gcc ttt ctg ctg atc ccc gac atc cag atg acc cag acc acc tcc agc         96
Ala Phe Leu Leu Ile Pro Asp Ile Gln Met Thr Gln Thr Thr Ser Ser           
            20                  25                  30                    

ctg agc gcc agc ctg ggc gac cgg gtg acc atc agc tgc cgg gcc agc        144
Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala Ser           
        35                  40                  45                        

cag gac atc agc aag tac ctg aac tgg tat cag cag aag ccc gac ggc        192
Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly           
    50                  55                  60                            

acc gtc aag ctg ctg atc tac cac acc agc cgg ctg cac agc ggc gtg        240
Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly Val           
65                  70                  75                  80            

ccc agc cgg ttt agc ggc agc ggc tcc ggc acc gac tac agc ctg acc        288
Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr           
                85                  90                  95                

atc tcc aac ctg gaa cag gaa gat atc gcc acc tac ttt tgc cag cag        336
Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln           
            100                 105                 110                   

ggc aac aca ctg ccc tac acc ttt ggc ggc gga aca aag ctg gaa atc        384
Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile           
        115                 120                 125                       

acc ggc agc acc tcc ggc agc ggc aag cct ggc agc ggc gag ggc agc        432
Thr Gly Ser Thr Ser Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser           
    130                 135                 140                           

acc aag ggc gag gtg aag ctg cag gaa agc ggc cct ggc ctg gtg gcc        480
Thr Lys Gly Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala           
145                 150                 155                 160           

ccc agc cag agc ctg agc gtg acc tgc acc gtg agc ggc gtg agc ctg        528
Pro Ser Gln Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu           
                165                 170                 175               

ccc gac tac ggc gtg agc tgg atc cgg cag ccc ccc agg aag ggc ctg        576
Pro Asp Tyr Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu           
            180                 185                 190                   

gaa tgg ctg ggc gtg atc tgg ggc agc gag acc acc tac tac aac agc        624
Glu Trp Leu Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser           
        195                 200                 205                       

gcc ctg aag agc cgg ctg acc atc atc aag gac aac agc aag agc cag        672
Ala Leu Lys Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln           
    210                 215                 220                           

gtg ttc ctg aag atg aac agc ctg cag acc gac gac acc gcc atc tac        720
Val Phe Leu Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr           
225                 230                 235                 240           

tac tgc gcc aag cac tac tac tac ggc ggc agc tac gcc atg gac tac        768
Tyr Cys Ala Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr           
                245                 250                 255               

tgg ggc cag ggc acc agc gtg acc gtg agc agc gga ggt ggt gga tcc        816
Trp Gly Gln Gly Thr Ser Val Thr Val Ser Ser Gly Gly Gly Gly Ser           
            260                 265                 270                   

gag gtg cag ctg cag cag tct ggg gct gag ctg gtg aag cct ggg gcc        864
Glu Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala           
        275                 280                 285                       

tca gtg aag atg tcc tgc aag gct tct ggc tac aca ttt acc agt tac        912
Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr           
    290                 295                 300                           

aat atg cac tgg gta aag cag aca cct gga cag ggc ctg gaa tgg att        960
Asn Met His Trp Val Lys Gln Thr Pro Gly Gln Gly Leu Glu Trp Ile           
305                 310                 315                 320           

gga gct att tat cca gga aat ggt gat act tcc tac aat cag aag ttc       1008
Gly Ala Ile Tyr Pro Gly Asn Gly Asp Thr Ser Tyr Asn Gln Lys Phe           
                325                 330                 335               

aaa ggc aag gcc aca ttg act gca gac aaa tcc tcc agc aca gcc tac       1056
Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr           
            340                 345                 350                   

atg cag ctc agc agc ctg aca tct gag gac tct gcg gac tat tac tgt       1104
Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Asp Tyr Tyr Cys           
        355                 360                 365                       

gca aga tct aat tat tac ggt agt agc tac tgg ttc ttc gat gtc tgg       1152
Ala Arg Ser Asn Tyr Tyr Gly Ser Ser Tyr Trp Phe Phe Asp Val Trp           
    370                 375                 380                           

ggc gca ggg acc acg gtc acc gtc tcc tca ggc agt act agc ggt ggt       1200
Gly Ala Gly Thr Thr Val Thr Val Ser Ser Gly Ser Thr Ser Gly Gly           
385                 390                 395                 400           

ggc tcc ggg ggc ggt tcc ggt ggg ggc ggc agc agc gac att gtg ctg       1248
Gly Ser Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Asp Ile Val Leu           
                405                 410                 415               

acc caa tct cca gct atc ctg tct gca tct cca ggg gag aag gtc aca       1296
Thr Gln Ser Pro Ala Ile Leu Ser Ala Ser Pro Gly Glu Lys Val Thr           
            420                 425                 430                   

atg act tgc agg gcc agc tca agt gta aat tac atg gac tgg tac cag       1344
Met Thr Cys Arg Ala Ser Ser Ser Val Asn Tyr Met Asp Trp Tyr Gln           
        435                 440                 445                       

aag aag cca gga tcc tcc ccc aaa ccc tgg att tat gcc aca tcc aac       1392
Lys Lys Pro Gly Ser Ser Pro Lys Pro Trp Ile Tyr Ala Thr Ser Asn           
    450                 455                 460                           

ctg gct tct gga gtc cct gct cgc ttc agt ggc agt ggg tct ggg acc       1440
Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr           
465                 470                 475                 480           

tct tac tct ctc aca atc agc aga gtg gag gct gaa gat gct gcc act       1488
Ser Tyr Ser Leu Thr Ile Ser Arg Val Glu Ala Glu Asp Ala Ala Thr           
                485                 490                 495               

tat tac tgc cag cag tgg agt ttt aat cca ccc acg ttc gga ggg ggg       1536
Tyr Tyr Cys Gln Gln Trp Ser Phe Asn Pro Pro Thr Phe Gly Gly Gly           
            500                 505                 510                   

acc aag ctg gaa ata aaa gag agc aag tac gga ccg ccc tgc ccc cct       1584
Thr Lys Leu Glu Ile Lys Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro           
        515                 520                 525                       

tgc cct gcc ccc gag ttc ctg ggc gga ccc agc gtg ttc ctg ttc ccc       1632
Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro           
    530                 535                 540                           

ccc aag ccc aag gac acc ctg atg atc agc cgg acc ccc gag gtg acc       1680
Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr           
545                 550                 555                 560           

tgc gtg gtg gtg gac gtg agc cag gaa gat ccc gag gtc cag ttc aat       1728
Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn           
                565                 570                 575               

tgg tac gtg gac ggc gtg gaa gtg cac aac gcc aag acc aag ccc aga       1776
Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg           
            580                 585                 590                   

gag gaa cag ttc aac agc acc tac cgg gtg gtg tct gtg ctg acc gtg       1824
Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val           
        595                 600                 605                       

ctg cac cag gac tgg ctg aac ggc aaa gaa tac aag tgc aag gtg tcc       1872
Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser           
    610                 615                 620                           

aac aag ggc ctg ccc agc agc atc gaa aag acc atc agc aag gcc aag       1920
Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys           
625                 630                 635                 640           

ggc cag cct cgc gag ccc cag gtg tac acc ctg cct ccc tcc cag gaa       1968
Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu           
                645                 650                 655               

gag atg acc aag aac cag gtg tcc ctg acc tgc ctg gtg aag ggc ttc       2016
Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe           
            660                 665                 670                   

tac ccc agc gac atc gcc gtg gag tgg gag agc aac ggc cag cct gag       2064
Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu           
        675                 680                 685                       

aac aac tac aag acc acc cct ccc gtg ctg gac agc gac ggc agc ttc       2112
Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe           
    690                 695                 700                           

ttc ctg tac agc cgg ctg acc gtg gac aag agc cgg tgg cag gaa ggc       2160
Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly           
705                 710                 715                 720           

aac gtc ttt agc tgc agc gtg atg cac gag gcc ctg cac aac cac tac       2208
Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr           
                725                 730                 735               

acc cag aag agc ctg agc ctg tcc ctg ggc aag atg ttc tgg gtg ctg       2256
Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Met Phe Trp Val Leu           
            740                 745                 750                   

gtg gtg gtg ggc ggg gtg ctg gcc tgc tac agc ctg ctg gtg aca gtg       2304
Val Val Val Gly Gly Val Leu Ala Cys Tyr Ser Leu Leu Val Thr Val           
        755                 760                 765                       

gcc ttc atc atc ttt tgg gtg cgg agc aag cgg agc aga ggc ggc cac       2352
Ala Phe Ile Ile Phe Trp Val Arg Ser Lys Arg Ser Arg Gly Gly His           
    770                 775                 780                           

agc gac tac atg aac atg acc ccc aga cgg cct ggc ccc acc cgg aag       2400
Ser Asp Tyr Met Asn Met Thr Pro Arg Arg Pro Gly Pro Thr Arg Lys           
785                 790                 795                 800           

cac tac cag ccc tac gcc cca ccc agg gac ttt gcc gcc tac aga agc       2448
His Tyr Gln Pro Tyr Ala Pro Pro Arg Asp Phe Ala Ala Tyr Arg Ser           
                805                 810                 815               

aaa cgg ggc aga aag aaa ctc ctg tat ata ttc aaa caa cca ttt atg       2496
Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met           
            820                 825                 830                   

aga cca gta caa act act caa gag gaa gat ggc tgt agc tgc cga ttt       2544
Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe           
        835                 840                 845                       

cca gaa gaa gaa gaa gga gga tgt gaa ctg cgg gtg aag ttc agc aga       2592
Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu Arg Val Lys Phe Ser Arg           
    850                 855                 860                           

agc gcc gac gcc cct gcc tac cag cag ggc cag aat cag ctg tac aac       2640
Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn           
865                 870                 875                 880           

gag ctg aac ctg ggc aga agg gaa gag tac gac gtc ctg gat aag cgg       2688
Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg           
                885                 890                 895               

aga ggc cgg gac cct gag atg ggc ggc aag cct cgg cgg aag aac ccc       2736
Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro           
            900                 905                 910                   

cag gaa ggc ctg tat aac gaa ctg cag aaa gac aag atg gcc gag gcc       2784
Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala           
        915                 920                 925                       

tac agc gag atc ggc atg aag ggc gag cgg agg cgg ggc aag ggc cac       2832
Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His           
    930                 935                 940                           

gac ggc ctg tat cag ggc ctg tcc acc gcc acc aag gat acc tac gac       2880
Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp           
945                 950                 955                 960           

gcc ctg cac atg cag gcc ctg ccc cca agg                               2910
Ala Leu His Met Gln Ala Leu Pro Pro Arg                                   
                965                 970                                   


<210>  9
<211>  970
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic Construct

<400>  9

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 Thr Thr Ser Ser 
            20                  25                  30          


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


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


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


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


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


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


Thr Gly Ser Thr Ser Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser 
    130                 135                 140                 


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


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


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


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


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


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


Tyr Cys Ala Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr 
                245                 250                 255     


Trp Gly Gln Gly Thr Ser Val Thr Val Ser Ser Gly Gly Gly Gly Ser 
            260                 265                 270         


Glu Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala 
        275                 280                 285             


Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 
    290                 295                 300                 


Asn Met His Trp Val Lys Gln Thr Pro Gly Gln Gly Leu Glu Trp Ile 
305                 310                 315                 320 


Gly Ala Ile Tyr Pro Gly Asn Gly Asp Thr Ser Tyr Asn Gln Lys Phe 
                325                 330                 335     


Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr 
            340                 345                 350         


Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Asp Tyr Tyr Cys 
        355                 360                 365             


Ala Arg Ser Asn Tyr Tyr Gly Ser Ser Tyr Trp Phe Phe Asp Val Trp 
    370                 375                 380                 


Gly Ala Gly Thr Thr Val Thr Val Ser Ser Gly Ser Thr Ser Gly Gly 
385                 390                 395                 400 


Gly Ser Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Asp Ile Val Leu 
                405                 410                 415     


Thr Gln Ser Pro Ala Ile Leu Ser Ala Ser Pro Gly Glu Lys Val Thr 
            420                 425                 430         


Met Thr Cys Arg Ala Ser Ser Ser Val Asn Tyr Met Asp Trp Tyr Gln 
        435                 440                 445             


Lys Lys Pro Gly Ser Ser Pro Lys Pro Trp Ile Tyr Ala Thr Ser Asn 
    450                 455                 460                 


Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr 
465                 470                 475                 480 


Ser Tyr Ser Leu Thr Ile Ser Arg Val Glu Ala Glu Asp Ala Ala Thr 
                485                 490                 495     


Tyr Tyr Cys Gln Gln Trp Ser Phe Asn Pro Pro Thr Phe Gly Gly Gly 
            500                 505                 510         


Thr Lys Leu Glu Ile Lys Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro 
        515                 520                 525             


Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro 
    530                 535                 540                 


Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr 
545                 550                 555                 560 


Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn 
                565                 570                 575     


Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg 
            580                 585                 590         


Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val 
        595                 600                 605             


Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser 
    610                 615                 620                 


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


Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu 
                645                 650                 655     


Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe 
            660                 665                 670         


Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu 
        675                 680                 685             


Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe 
    690                 695                 700                 


Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly 
705                 710                 715                 720 


Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr 
                725                 730                 735     


Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Met Phe Trp Val Leu 
            740                 745                 750         


Val Val Val Gly Gly Val Leu Ala Cys Tyr Ser Leu Leu Val Thr Val 
        755                 760                 765             


Ala Phe Ile Ile Phe Trp Val Arg Ser Lys Arg Ser Arg Gly Gly His 
    770                 775                 780                 


Ser Asp Tyr Met Asn Met Thr Pro Arg Arg Pro Gly Pro Thr Arg Lys 
785                 790                 795                 800 


His Tyr Gln Pro Tyr Ala Pro Pro Arg Asp Phe Ala Ala Tyr Arg Ser 
                805                 810                 815     


Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met 
            820                 825                 830         


Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe 
        835                 840                 845             


Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu Arg Val Lys Phe Ser Arg 
    850                 855                 860                 


Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn 
865                 870                 875                 880 


Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg 
                885                 890                 895     


Arg Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro 
            900                 905                 910         


Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala 
        915                 920                 925             


Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His 
    930                 935                 940                 


Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp 
945                 950                 955                 960 


Ala Leu His Met Gln Ala Leu Pro Pro Arg 
                965                 970 


<210>  10
<211>  3402
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  GMCSFRss-CD19scFv-Gly4serlinker-CD20scFv-CD8alphaHinge-CD8alphaTM
       -41BB-CD3Zeta-T2A-EGFRt

<400>  10
atgctgctgc tggtgaccag cctgctgctg tgcgagctgc cccaccccgc ctttctgctg       60

atccccgaca tccagatgac ccagaccacc tccagcctga gcgccagcct gggcgaccgg      120

gtgaccatca gctgccgggc cagccaggac atcagcaagt acctgaactg gtatcagcag      180

aagcccgacg gcaccgtcaa gctgctgatc taccacacca gccggctgca cagcggcgtg      240

cccagccggt ttagcggcag cggctccggc accgactaca gcctgaccat ctccaacctg      300

gaacaggaag atatcgccac ctacttttgc cagcagggca acacactgcc ctacaccttt      360

ggcggcggaa caaagctgga aatcaccggc agcacctccg gcagcggcaa gcctggcagc      420

ggcgagggca gcaccaaggg cgaggtgaag ctgcaggaaa gcggccctgg cctggtggcc      480

cccagccaga gcctgagcgt gacctgcacc gtgagcggcg tgagcctgcc cgactacggc      540

gtgagctgga tccggcagcc ccccaggaag ggcctggaat ggctgggcgt gatctggggc      600

agcgagacca cctactacaa cagcgccctg aagagccggc tgaccatcat caaggacaac      660

agcaagagcc aggtgttcct gaagatgaac agcctgcaga ccgacgacac cgccatctac      720

tactgcgcca agcactacta ctacggcggc agctacgcca tggactactg gggccagggc      780

accagcgtga ccgtgagcag cggaggtggt ggatccgagg tgcagctgca gcagtctggg      840

gctgagctgg tgaagcctgg ggcctcagtg aagatgtcct gcaaggcttc tggctacaca      900

tttaccagtt acaatatgca ctgggtaaag cagacacctg gacagggcct ggaatggatt      960

ggagctattt atccaggaaa tggtgatact tcctacaatc agaagttcaa aggcaaggcc     1020

acattgactg cagacaaatc ctccagcaca gcctacatgc agctcagcag cctgacatct     1080

gaggactctg cggactatta ctgtgcaaga tctaattatt acggtagtag ctactggttc     1140

ttcgatgtct ggggcgcagg gaccacggtc accgtctcct caggcagtac tagcggtggt     1200

ggctccgggg gcggttccgg tgggggcggc agcagcgaca ttgtgctgac ccaatctcca     1260

gctatcctgt ctgcatctcc aggggagaag gtcacaatga cttgcagggc cagctcaagt     1320

gtaaattaca tggactggta ccagaagaag ccaggatcct cccccaaacc ctggatttat     1380

gccacatcca acctggcttc tggagtccct gctcgcttca gtggcagtgg gtctgggacc     1440

tcttactctc tcacaatcag cagagtggag gctgaagatg ctgccactta ttactgccag     1500

cagtggagtt ttaatccacc cacgttcgga ggggggacca agctggaaat aaaagagagc     1560

aagtacggac cgccctgccc cccttgccct aagcctacca ccacccctgc ccctagacct     1620

ccaacacccg ccccaacaat cgccagccag cctctgtctc tgaggcccga ggcttgtaga     1680

ccagctgctg gcggagccgt gcacaccaga ggactggatt tcgcctgcga catctacatc     1740

tgggcccctc tggccggcac atgtggcgtg ctgctgctga gcctcgtgat caccaagcgg     1800

ggcagaaaga aactgctgta catctttaag cagcccttca tgcggcccgt gcagaccacc     1860

caggaagagg acggctgctc ctgcagattc cccgaggaag aagaaggcgg ctgcgagctg     1920

agagtgaagt tcagcagatc cgccgacgcc cctgcctacc agcagggaca gaaccagctg     1980

tacaacgagc tgaacctggg cagacgggaa gagtacgacg tgctggacaa gcggagaggc     2040

cgggaccctg agatgggcgg aaagcccaga agaaagaacc cccaggaagg cctgtataac     2100

gaactgcaga aagacaagat ggccgaggcc tacagcgaga tcggaatgaa gggcgagcgg     2160

agaagaggca agggccacga tggcctgtac cagggcctga gcaccgccac caaggacacc     2220

tatgacgccc tgcacatgca ggccctgcct ccaagactcg agggcggcgg agagggcaga     2280

ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctaggat gcttctcctg     2340

gtgacaagcc ttctgctctg tgagttacca cacccagcat tcctcctgat cccacgcaaa     2400

gtgtgtaacg gaataggtat tggtgaattt aaagactcac tctccataaa tgctacgaat     2460

attaaacact tcaaaaactg cacctccatc agtggcgatc tccacatcct gccggtggca     2520

tttaggggtg actccttcac acatactcct cctctggatc cacaggaact ggatattctg     2580

aaaaccgtaa aggaaatcac agggtttttg ctgattcagg cttggcctga aaacaggacg     2640

gacctccatg cctttgagaa cctagaaatc atacgcggca ggaccaagca acatggtcag     2700

ttttctcttg cagtcgtcag cctgaacata acatccttgg gattacgctc cctcaaggag     2760

ataagtgatg gagatgtgat aatttcagga aacaaaaatt tgtgctatgc aaatacaata     2820

aactggaaaa aactgtttgg gacctccggt cagaaaacca aaattataag caacagaggt     2880

gaaaacagct gcaaggccac aggccaggtc tgccatgcct tgtgctcccc cgagggctgc     2940

tggggcccgg agcccaggga ctgcgtctct tgccggaatg tcagccgagg cagggaatgc     3000

gtggacaagt gcaaccttct ggagggtgag ccaagggagt ttgtggagaa ctctgagtgc     3060

atacagtgcc acccagagtg cctgcctcag gccatgaaca tcacctgcac aggacgggga     3120

ccagacaact gtatccagtg tgcccactac attgacggcc cccactgcgt caagacctgc     3180

ccggcaggag tcatgggaga aaacaacacc ctggtctgga agtacgcaga cgccggccat     3240

gtgtgccacc tgtgccatcc aaactgcacc tacggatgca ctgggccagg tcttgaaggc     3300

tgtccaacga atgggcctaa gatcccgtcc atcgccactg ggatggtggg ggccctcctc     3360

ttgctgctgg tggtggccct ggggatcggc ctcttcatgt ga                        3402


<210>  11
<211>  3402
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  GMCSFRss-CD19scFv-Gly4serlinker-CD20scFv-CD8alphaHinge-CD8alphaTM
       -41BB-CD3Zeta-T2A-EGFRt


<220>
<221>  CDS
<222>  (1)..(3402)

<400>  11
atg ctg ctg ctg gtg acc agc ctg ctg ctg tgc gag ctg ccc cac ccc         48
Met Leu Leu Leu Val Thr Ser Leu Leu Leu Cys Glu Leu Pro His Pro           
1               5                   10                  15                

gcc ttt ctg ctg atc ccc gac atc cag atg acc cag acc acc tcc agc         96
Ala Phe Leu Leu Ile Pro Asp Ile Gln Met Thr Gln Thr Thr Ser Ser           
            20                  25                  30                    

ctg agc gcc agc ctg ggc gac cgg gtg acc atc agc tgc cgg gcc agc        144
Leu Ser Ala Ser Leu Gly Asp Arg Val Thr Ile Ser Cys Arg Ala Ser           
        35                  40                  45                        

cag gac atc agc aag tac ctg aac tgg tat cag cag aag ccc gac ggc        192
Gln Asp Ile Ser Lys Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly           
    50                  55                  60                            

acc gtc aag ctg ctg atc tac cac acc agc cgg ctg cac agc ggc gtg        240
Thr Val Lys Leu Leu Ile Tyr His Thr Ser Arg Leu His Ser Gly Val           
65                  70                  75                  80            

ccc agc cgg ttt agc ggc agc ggc tcc ggc acc gac tac agc ctg acc        288
Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr           
                85                  90                  95                

atc tcc aac ctg gaa cag gaa gat atc gcc acc tac ttt tgc cag cag        336
Ile Ser Asn Leu Glu Gln Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln           
            100                 105                 110                   

ggc aac aca ctg ccc tac acc ttt ggc ggc gga aca aag ctg gaa atc        384
Gly Asn Thr Leu Pro Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile           
        115                 120                 125                       

acc ggc agc acc tcc ggc agc ggc aag cct ggc agc ggc gag ggc agc        432
Thr Gly Ser Thr Ser Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser           
    130                 135                 140                           

acc aag ggc gag gtg aag ctg cag gaa agc ggc cct ggc ctg gtg gcc        480
Thr Lys Gly Glu Val Lys Leu Gln Glu Ser Gly Pro Gly Leu Val Ala           
145                 150                 155                 160           

ccc agc cag agc ctg agc gtg acc tgc acc gtg agc ggc gtg agc ctg        528
Pro Ser Gln Ser Leu Ser Val Thr Cys Thr Val Ser Gly Val Ser Leu           
                165                 170                 175               

ccc gac tac ggc gtg agc tgg atc cgg cag ccc ccc agg aag ggc ctg        576
Pro Asp Tyr Gly Val Ser Trp Ile Arg Gln Pro Pro Arg Lys Gly Leu           
            180                 185                 190                   

gaa tgg ctg ggc gtg atc tgg ggc agc gag acc acc tac tac aac agc        624
Glu Trp Leu Gly Val Ile Trp Gly Ser Glu Thr Thr Tyr Tyr Asn Ser           
        195                 200                 205                       

gcc ctg aag agc cgg ctg acc atc atc aag gac aac agc aag agc cag        672
Ala Leu Lys Ser Arg Leu Thr Ile Ile Lys Asp Asn Ser Lys Ser Gln           
    210                 215                 220                           

gtg ttc ctg aag atg aac agc ctg cag acc gac gac acc gcc atc tac        720
Val Phe Leu Lys Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr           
225                 230                 235                 240           

tac tgc gcc aag cac tac tac tac ggc ggc agc tac gcc atg gac tac        768
Tyr Cys Ala Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr           
                245                 250                 255               

tgg ggc cag ggc acc agc gtg acc gtg agc agc gga ggt ggt gga tcc        816
Trp Gly Gln Gly Thr Ser Val Thr Val Ser Ser Gly Gly Gly Gly Ser           
            260                 265                 270                   

gag gtg cag ctg cag cag tct ggg gct gag ctg gtg aag cct ggg gcc        864
Glu Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala           
        275                 280                 285                       

tca gtg aag atg tcc tgc aag gct tct ggc tac aca ttt acc agt tac        912
Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr           
    290                 295                 300                           

aat atg cac tgg gta aag cag aca cct gga cag ggc ctg gaa tgg att        960
Asn Met His Trp Val Lys Gln Thr Pro Gly Gln Gly Leu Glu Trp Ile           
305                 310                 315                 320           

gga gct att tat cca gga aat ggt gat act tcc tac aat cag aag ttc       1008
Gly Ala Ile Tyr Pro Gly Asn Gly Asp Thr Ser Tyr Asn Gln Lys Phe           
                325                 330                 335               

aaa ggc aag gcc aca ttg act gca gac aaa tcc tcc agc aca gcc tac       1056
Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr           
            340                 345                 350                   

atg cag ctc agc agc ctg aca tct gag gac tct gcg gac tat tac tgt       1104
Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Asp Tyr Tyr Cys           
        355                 360                 365                       

gca aga tct aat tat tac ggt agt agc tac tgg ttc ttc gat gtc tgg       1152
Ala Arg Ser Asn Tyr Tyr Gly Ser Ser Tyr Trp Phe Phe Asp Val Trp           
    370                 375                 380                           

ggc gca ggg acc acg gtc acc gtc tcc tca ggc agt act agc ggt ggt       1200
Gly Ala Gly Thr Thr Val Thr Val Ser Ser Gly Ser Thr Ser Gly Gly           
385                 390                 395                 400           

ggc tcc ggg ggc ggt tcc ggt ggg ggc ggc agc agc gac att gtg ctg       1248
Gly Ser Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Asp Ile Val Leu           
                405                 410                 415               

acc caa tct cca gct atc ctg tct gca tct cca ggg gag aag gtc aca       1296
Thr Gln Ser Pro Ala Ile Leu Ser Ala Ser Pro Gly Glu Lys Val Thr           
            420                 425                 430                   

atg act tgc agg gcc agc tca agt gta aat tac atg gac tgg tac cag       1344
Met Thr Cys Arg Ala Ser Ser Ser Val Asn Tyr Met Asp Trp Tyr Gln           
        435                 440                 445                       

aag aag cca gga tcc tcc ccc aaa ccc tgg att tat gcc aca tcc aac       1392
Lys Lys Pro Gly Ser Ser Pro Lys Pro Trp Ile Tyr Ala Thr Ser Asn           
    450                 455                 460                           

ctg gct tct gga gtc cct gct cgc ttc agt ggc agt ggg tct ggg acc       1440
Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr           
465                 470                 475                 480           

tct tac tct ctc aca atc agc aga gtg gag gct gaa gat gct gcc act       1488
Ser Tyr Ser Leu Thr Ile Ser Arg Val Glu Ala Glu Asp Ala Ala Thr           
                485                 490                 495               

tat tac tgc cag cag tgg agt ttt aat cca ccc acg ttc gga ggg ggg       1536
Tyr Tyr Cys Gln Gln Trp Ser Phe Asn Pro Pro Thr Phe Gly Gly Gly           
            500                 505                 510                   

acc aag ctg gaa ata aaa gag agc aag tac gga ccg ccc tgc ccc cct       1584
Thr Lys Leu Glu Ile Lys Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro           
        515                 520                 525                       

tgc cct aag cct acc acc acc cct gcc cct aga cct cca aca ccc gcc       1632
Cys Pro Lys Pro Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala           
    530                 535                 540                           

cca aca atc gcc agc cag cct ctg tct ctg agg ccc gag gct tgt aga       1680
Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg           
545                 550                 555                 560           

cca gct gct ggc gga gcc gtg cac acc aga gga ctg gat ttc gcc tgc       1728
Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys           
                565                 570                 575               

gac atc tac atc tgg gcc cct ctg gcc ggc aca tgt ggc gtg ctg ctg       1776
Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu           
            580                 585                 590                   

ctg agc ctc gtg atc acc aag cgg ggc aga aag aaa ctg ctg tac atc       1824
Leu Ser Leu Val Ile Thr Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile           
        595                 600                 605                       

ttt aag cag ccc ttc atg cgg ccc gtg cag acc acc cag gaa gag gac       1872
Phe Lys Gln Pro Phe Met Arg Pro Val Gln Thr Thr Gln Glu Glu Asp           
    610                 615                 620                           

ggc tgc tcc tgc aga ttc ccc gag gaa gaa gaa ggc ggc tgc gag ctg       1920
Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu           
625                 630                 635                 640           

aga gtg aag ttc agc aga tcc gcc gac gcc cct gcc tac cag cag gga       1968
Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly           
                645                 650                 655               

cag aac cag ctg tac aac gag ctg aac ctg ggc aga cgg gaa gag tac       2016
Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr           
            660                 665                 670                   

gac gtg ctg gac aag cgg aga ggc cgg gac cct gag atg ggc gga aag       2064
Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys           
        675                 680                 685                       

ccc aga aga aag aac ccc cag gaa ggc ctg tat aac gaa ctg cag aaa       2112
Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys           
    690                 695                 700                           

gac aag atg gcc gag gcc tac agc gag atc gga atg aag ggc gag cgg       2160
Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg           
705                 710                 715                 720           

aga aga ggc aag ggc cac gat ggc ctg tac cag ggc ctg agc acc gcc       2208
Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala           
                725                 730                 735               

acc aag gac acc tat gac gcc ctg cac atg cag gcc ctg cct cca aga       2256
Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg           
            740                 745                 750                   

ctc gag ggc ggc gga gag ggc aga gga agt ctt cta aca tgc ggt gac       2304
Leu Glu Gly Gly Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly Asp           
        755                 760                 765                       

gtg gag gag aat ccc ggc cct agg atg ctt ctc ctg gtg aca agc ctt       2352
Val Glu Glu Asn Pro Gly Pro Arg Met Leu Leu Leu Val Thr Ser Leu           
    770                 775                 780                           

ctg ctc tgt gag tta cca cac cca gca ttc ctc ctg atc cca cgc aaa       2400
Leu Leu Cys Glu Leu Pro His Pro Ala Phe Leu Leu Ile Pro Arg Lys           
785                 790                 795                 800           

gtg tgt aac gga ata ggt att ggt gaa ttt aaa gac tca ctc tcc ata       2448
Val Cys Asn Gly Ile Gly Ile Gly Glu Phe Lys Asp Ser Leu Ser Ile           
                805                 810                 815               

aat gct acg aat att aaa cac ttc aaa aac tgc acc tcc atc agt ggc       2496
Asn Ala Thr Asn Ile Lys His Phe Lys Asn Cys Thr Ser Ile Ser Gly           
            820                 825                 830                   

gat ctc cac atc ctg ccg gtg gca ttt agg ggt gac tcc ttc aca cat       2544
Asp Leu His Ile Leu Pro Val Ala Phe Arg Gly Asp Ser Phe Thr His           
        835                 840                 845                       

act cct cct ctg gat cca cag gaa ctg gat att ctg aaa acc gta aag       2592
Thr Pro Pro Leu Asp Pro Gln Glu Leu Asp Ile Leu Lys Thr Val Lys           
    850                 855                 860                           

gaa atc aca ggg ttt ttg ctg att cag gct tgg cct gaa aac agg acg       2640
Glu Ile Thr Gly Phe Leu Leu Ile Gln Ala Trp Pro Glu Asn Arg Thr           
865                 870                 875                 880           

gac ctc cat gcc ttt gag aac cta gaa atc ata cgc ggc agg acc aag       2688
Asp Leu His Ala Phe Glu Asn Leu Glu Ile Ile Arg Gly Arg Thr Lys           
                885                 890                 895               

caa cat ggt cag ttt tct ctt gca gtc gtc agc ctg aac ata aca tcc       2736
Gln His Gly Gln Phe Ser Leu Ala Val Val Ser Leu Asn Ile Thr Ser           
            900                 905                 910                   

ttg gga tta cgc tcc ctc aag gag ata agt gat gga gat gtg ata att       2784
Leu Gly Leu Arg Ser Leu Lys Glu Ile Ser Asp Gly Asp Val Ile Ile           
        915                 920                 925                       

tca gga aac aaa aat ttg tgc tat gca aat aca ata aac tgg aaa aaa       2832
Ser Gly Asn Lys Asn Leu Cys Tyr Ala Asn Thr Ile Asn Trp Lys Lys           
    930                 935                 940                           

ctg ttt ggg acc tcc ggt cag aaa acc aaa att ata agc aac aga ggt       2880
Leu Phe Gly Thr Ser Gly Gln Lys Thr Lys Ile Ile Ser Asn Arg Gly           
945                 950                 955                 960           

gaa aac agc tgc aag gcc aca ggc cag gtc tgc cat gcc ttg tgc tcc       2928
Glu Asn Ser Cys Lys Ala Thr Gly Gln Val Cys His Ala Leu Cys Ser           
                965                 970                 975               

ccc gag ggc tgc tgg ggc ccg gag ccc agg gac tgc gtc tct tgc cgg       2976
Pro Glu Gly Cys Trp Gly Pro Glu Pro Arg Asp Cys Val Ser Cys Arg           
            980                 985                 990                   

aat gtc agc cga ggc agg gaa tgc  gtg gac aag tgc aac  ctt ctg gag     3024
Asn Val Ser Arg Gly Arg Glu Cys  Val Asp Lys Cys Asn  Leu Leu Glu         
        995                 1000                 1005                     

ggt gag  cca agg gag ttt gtg  gag aac tct gag tgc  ata cag tgc        3069
Gly Glu  Pro Arg Glu Phe Val  Glu Asn Ser Glu Cys  Ile Gln Cys            
    1010                 1015                 1020                        

cac cca  gag tgc ctg cct cag  gcc atg aac atc acc  tgc aca gga        3114
His Pro  Glu Cys Leu Pro Gln  Ala Met Asn Ile Thr  Cys Thr Gly            
    1025                 1030                 1035                        

cgg gga  cca gac aac tgt atc  cag tgt gcc cac tac  att gac ggc        3159
Arg Gly  Pro Asp Asn Cys Ile  Gln Cys Ala His Tyr  Ile Asp Gly            
    1040                 1045                 1050                        

ccc cac  tgc gtc aag acc tgc  ccg gca gga gtc atg  gga gaa aac        3204
Pro His  Cys Val Lys Thr Cys  Pro Ala Gly Val Met  Gly Glu Asn            
    1055                 1060                 1065                        

aac acc  ctg gtc tgg aag tac  gca gac gcc ggc cat  gtg tgc cac        3249
Asn Thr  Leu Val Trp Lys Tyr  Ala Asp Ala Gly His  Val Cys His            
    1070                 1075                 1080                        

ctg tgc  cat cca aac tgc acc  tac gga tgc act ggg  cca ggt ctt        3294
Leu Cys  His Pro Asn Cys Thr  Tyr Gly Cys Thr Gly  Pro Gly Leu            
    1085                 1090                 1095                        

gaa ggc  tgt cca acg aat ggg  cct aag atc ccg tcc  atc gcc act        3339
Glu Gly  Cys Pro Thr Asn Gly  Pro Lys Ile Pro Ser  Ile Ala Thr            
    1100                 1105                 1110                        

ggg atg  gtg ggg gcc ctc ctc  ttg ctg ctg gtg gtg  gcc ctg ggg        3384
Gly Met  Val Gly Ala Leu Leu  Leu Leu Leu Val Val  Ala Leu Gly            
    1115                 1120                 1125                        

atc ggc  ctc ttc atg tga                                              3402
Ile Gly  Leu Phe Met                                                      
    1130                                                                  


<210>  12
<211>  1133
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic Construct

<400>  12

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 Thr Thr Ser Ser 
            20                  25                  30          


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


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


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


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


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


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


Thr Gly Ser Thr Ser Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser 
    130                 135                 140                 


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


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


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


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


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


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


Tyr Cys Ala Lys His Tyr Tyr Tyr Gly Gly Ser Tyr Ala Met Asp Tyr 
                245                 250                 255     


Trp Gly Gln Gly Thr Ser Val Thr Val Ser Ser Gly Gly Gly Gly Ser 
            260                 265                 270         


Glu Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Lys Pro Gly Ala 
        275                 280                 285             


Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 
    290                 295                 300                 


Asn Met His Trp Val Lys Gln Thr Pro Gly Gln Gly Leu Glu Trp Ile 
305                 310                 315                 320 


Gly Ala Ile Tyr Pro Gly Asn Gly Asp Thr Ser Tyr Asn Gln Lys Phe 
                325                 330                 335     


Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr 
            340                 345                 350         


Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Asp Tyr Tyr Cys 
        355                 360                 365             


Ala Arg Ser Asn Tyr Tyr Gly Ser Ser Tyr Trp Phe Phe Asp Val Trp 
    370                 375                 380                 


Gly Ala Gly Thr Thr Val Thr Val Ser Ser Gly Ser Thr Ser Gly Gly 
385                 390                 395                 400 


Gly Ser Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Asp Ile Val Leu 
                405                 410                 415     


Thr Gln Ser Pro Ala Ile Leu Ser Ala Ser Pro Gly Glu Lys Val Thr 
            420                 425                 430         


Met Thr Cys Arg Ala Ser Ser Ser Val Asn Tyr Met Asp Trp Tyr Gln 
        435                 440                 445             


Lys Lys Pro Gly Ser Ser Pro Lys Pro Trp Ile Tyr Ala Thr Ser Asn 
    450                 455                 460                 


Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser Gly Ser Gly Thr 
465                 470                 475                 480 


Ser Tyr Ser Leu Thr Ile Ser Arg Val Glu Ala Glu Asp Ala Ala Thr 
                485                 490                 495     


Tyr Tyr Cys Gln Gln Trp Ser Phe Asn Pro Pro Thr Phe Gly Gly Gly 
            500                 505                 510         


Thr Lys Leu Glu Ile Lys Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro 
        515                 520                 525             


Cys Pro Lys Pro Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala 
    530                 535                 540                 


Pro Thr Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg 
545                 550                 555                 560 


Pro Ala Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys 
                565                 570                 575     


Asp Ile Tyr Ile Trp Ala Pro Leu Ala Gly Thr Cys Gly Val Leu Leu 
            580                 585                 590         


Leu Ser Leu Val Ile Thr Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile 
        595                 600                 605             


Phe Lys Gln Pro Phe Met Arg Pro Val Gln Thr Thr Gln Glu Glu Asp 
    610                 615                 620                 


Gly Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu 
625                 630                 635                 640 


Arg Val Lys Phe Ser Arg Ser Ala Asp Ala Pro Ala Tyr Gln Gln Gly 
                645                 650                 655     


Gln Asn Gln Leu Tyr Asn Glu Leu Asn Leu Gly Arg Arg Glu Glu Tyr 
            660                 665                 670         


Asp Val Leu Asp Lys Arg Arg Gly Arg Asp Pro Glu Met Gly Gly Lys 
        675                 680                 685             


Pro Arg Arg Lys Asn Pro Gln Glu Gly Leu Tyr Asn Glu Leu Gln Lys 
    690                 695                 700                 


Asp Lys Met Ala Glu Ala Tyr Ser Glu Ile Gly Met Lys Gly Glu Arg 
705                 710                 715                 720 


Arg Arg Gly Lys Gly His Asp Gly Leu Tyr Gln Gly Leu Ser Thr Ala 
                725                 730                 735     


Thr Lys Asp Thr Tyr Asp Ala Leu His Met Gln Ala Leu Pro Pro Arg 
            740                 745                 750         


Leu Glu Gly Gly Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly Asp 
        755                 760                 765             


Val Glu Glu Asn Pro Gly Pro Arg Met Leu Leu Leu Val Thr Ser Leu 
    770                 775                 780                 


Leu Leu Cys Glu Leu Pro His Pro Ala Phe Leu Leu Ile Pro Arg Lys 
785                 790                 795                 800 


Val Cys Asn Gly Ile Gly Ile Gly Glu Phe Lys Asp Ser Leu Ser Ile 
                805                 810                 815     


Asn Ala Thr Asn Ile Lys His Phe Lys Asn Cys Thr Ser Ile Ser Gly 
            820                 825                 830         


Asp Leu His Ile Leu Pro Val Ala Phe Arg Gly Asp Ser Phe Thr His 
        835                 840                 845             


Thr Pro Pro Leu Asp Pro Gln Glu Leu Asp Ile Leu Lys Thr Val Lys 
    850                 855                 860                 


Glu Ile Thr Gly Phe Leu Leu Ile Gln Ala Trp Pro Glu Asn Arg Thr 
865                 870                 875                 880 


Asp Leu His Ala Phe Glu Asn Leu Glu Ile Ile Arg Gly Arg Thr Lys 
                885                 890                 895     


Gln His Gly Gln Phe Ser Leu Ala Val Val Ser Leu Asn Ile Thr Ser 
            900                 905                 910         


Leu Gly Leu Arg Ser Leu Lys Glu Ile Ser Asp Gly Asp Val Ile Ile 
        915                 920                 925             


Ser Gly Asn Lys Asn Leu Cys Tyr Ala Asn Thr Ile Asn Trp Lys Lys 
    930                 935                 940                 


Leu Phe Gly Thr Ser Gly Gln Lys Thr Lys Ile Ile Ser Asn Arg Gly 
945                 950                 955                 960 


Glu Asn Ser Cys Lys Ala Thr Gly Gln Val Cys His Ala Leu Cys Ser 
                965                 970                 975     


Pro Glu Gly Cys Trp Gly Pro Glu Pro Arg Asp Cys Val Ser Cys Arg 
            980                 985                 990         


Asn Val Ser Arg Gly Arg Glu Cys  Val Asp Lys Cys Asn  Leu Leu Glu 
        995                 1000                 1005             


Gly Glu  Pro Arg Glu Phe Val  Glu Asn Ser Glu Cys  Ile Gln Cys 
    1010                 1015                 1020             


His Pro  Glu Cys Leu Pro Gln  Ala Met Asn Ile Thr  Cys Thr Gly 
    1025                 1030                 1035             


Arg Gly  Pro Asp Asn Cys Ile  Gln Cys Ala His Tyr  Ile Asp Gly 
    1040                 1045                 1050             


Pro His  Cys Val Lys Thr Cys  Pro Ala Gly Val Met  Gly Glu Asn 
    1055                 1060                 1065             


Asn Thr  Leu Val Trp Lys Tyr  Ala Asp Ala Gly His  Val Cys His 
    1070                 1075                 1080             


Leu Cys  His Pro Asn Cys Thr  Tyr Gly Cys Thr Gly  Pro Gly Leu 
    1085                 1090                 1095             


Glu Gly  Cys Pro Thr Asn Gly  Pro Lys Ile Pro Ser  Ile Ala Thr 
    1100                 1105                 1110             


Gly Met  Val Gly Ala Leu Leu  Leu Leu Leu Val Val  Ala Leu Gly 
    1115                 1120                 1125             


Ile Gly  Leu Phe Met 
    1130             


<210>  13
<211>  1146
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  T2A-EGFRt

<400>  13
ctcgagggcg gcggagaggg cagaggaagt cttctaacat gcggtgacgt ggaggagaat       60

cccggcccta ggatgcttct cctggtgaca agccttctgc tctgtgagtt accacaccca      120

gcattcctcc tgatcccacg caaagtgtgt aacggaatag gtattggtga atttaaagac      180

tcactctcca taaatgctac gaatattaaa cacttcaaaa actgcacctc catcagtggc      240

gatctccaca tcctgccggt ggcatttagg ggtgactcct tcacacatac tcctcctctg      300

gatccacagg aactggatat tctgaaaacc gtaaaggaaa tcacagggtt tttgctgatt      360

caggcttggc ctgaaaacag gacggacctc catgcctttg agaacctaga aatcatacgc      420

ggcaggacca agcaacatgg tcagttttct cttgcagtcg tcagcctgaa cataacatcc      480

ttgggattac gctccctcaa ggagataagt gatggagatg tgataatttc aggaaacaaa      540

aatttgtgct atgcaaatac aataaactgg aaaaaactgt ttgggacctc cggtcagaaa      600

accaaaatta taagcaacag aggtgaaaac agctgcaagg ccacaggcca ggtctgccat      660

gccttgtgct cccccgaggg ctgctggggc ccggagccca gggactgcgt ctcttgccgg      720

aatgtcagcc gaggcaggga atgcgtggac aagtgcaacc ttctggaggg tgagccaagg      780

gagtttgtgg agaactctga gtgcatacag tgccacccag agtgcctgcc tcaggccatg      840

aacatcacct gcacaggacg gggaccagac aactgtatcc agtgtgccca ctacattgac      900

ggcccccact gcgtcaagac ctgcccggca ggagtcatgg gagaaaacaa caccctggtc      960

tggaagtacg cagacgccgg ccatgtgtgc cacctgtgcc atccaaactg cacctacgga     1020

tgcactgggc caggtcttga aggctgtcca acgaatgggc ctaagatccc gtccatcgcc     1080

actgggatgg tgggggccct cctcttgctg ctggtggtgg ccctggggat cggcctcttc     1140

atgtga                                                                1146


<210>  14
<211>  1146
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  T2A-EGFRt


<220>
<221>  CDS
<222>  (1)..(1146)

<400>  14
ctc gag ggc ggc gga gag ggc aga gga agt ctt cta aca tgc ggt gac         48
Leu Glu Gly Gly Gly Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly Asp           
1               5                   10                  15                

gtg gag gag aat ccc ggc cct agg atg ctt ctc ctg gtg aca agc ctt         96
Val Glu Glu Asn Pro Gly Pro Arg Met Leu Leu Leu Val Thr Ser Leu           
            20                  25                  30                    

ctg ctc tgt gag tta cca cac cca gca ttc ctc ctg atc cca cgc aaa        144
Leu Leu Cys Glu Leu Pro His Pro Ala Phe Leu Leu Ile Pro Arg Lys           
        35                  40                  45                        

gtg tgt aac gga ata ggt att ggt gaa ttt aaa gac tca ctc tcc ata        192
Val Cys Asn Gly Ile Gly Ile Gly Glu Phe Lys Asp Ser Leu Ser Ile           
    50                  55                  60                            

aat gct acg aat att aaa cac ttc aaa aac tgc acc tcc atc agt ggc        240
Asn Ala Thr Asn Ile Lys His Phe Lys Asn Cys Thr Ser Ile Ser Gly           
65                  70                  75                  80            

gat ctc cac atc ctg ccg gtg gca ttt agg ggt gac tcc ttc aca cat        288
Asp Leu His Ile Leu Pro Val Ala Phe Arg Gly Asp Ser Phe Thr His           
                85                  90                  95                

act cct cct ctg gat cca cag gaa ctg gat att ctg aaa acc gta aag        336
Thr Pro Pro Leu Asp Pro Gln Glu Leu Asp Ile Leu Lys Thr Val Lys           
            100                 105                 110                   

gaa atc aca ggg ttt ttg ctg att cag gct tgg cct gaa aac agg acg        384
Glu Ile Thr Gly Phe Leu Leu Ile Gln Ala Trp Pro Glu Asn Arg Thr           
        115                 120                 125                       

gac ctc cat gcc ttt gag aac cta gaa atc ata cgc ggc agg acc aag        432
Asp Leu His Ala Phe Glu Asn Leu Glu Ile Ile Arg Gly Arg Thr Lys           
    130                 135                 140                           

caa cat ggt cag ttt tct ctt gca gtc gtc agc ctg aac ata aca tcc        480
Gln His Gly Gln Phe Ser Leu Ala Val Val Ser Leu Asn Ile Thr Ser           
145                 150                 155                 160           

ttg gga tta cgc tcc ctc aag gag ata agt gat gga gat gtg ata att        528
Leu Gly Leu Arg Ser Leu Lys Glu Ile Ser Asp Gly Asp Val Ile Ile           
                165                 170                 175               

tca gga aac aaa aat ttg tgc tat gca aat aca ata aac tgg aaa aaa        576
Ser Gly Asn Lys Asn Leu Cys Tyr Ala Asn Thr Ile Asn Trp Lys Lys           
            180                 185                 190                   

ctg ttt ggg acc tcc ggt cag aaa acc aaa att ata agc aac aga ggt        624
Leu Phe Gly Thr Ser Gly Gln Lys Thr Lys Ile Ile Ser Asn Arg Gly           
        195                 200                 205                       

gaa aac agc tgc aag gcc aca ggc cag gtc tgc cat gcc ttg tgc tcc        672
Glu Asn Ser Cys Lys Ala Thr Gly Gln Val Cys His Ala Leu Cys Ser           
    210                 215                 220                           

ccc gag ggc tgc tgg ggc ccg gag ccc agg gac tgc gtc tct tgc cgg        720
Pro Glu Gly Cys Trp Gly Pro Glu Pro Arg Asp Cys Val Ser Cys Arg           
225                 230                 235                 240           

aat gtc agc cga ggc agg gaa tgc gtg gac aag tgc aac ctt ctg gag        768
Asn Val Ser Arg Gly Arg Glu Cys Val Asp Lys Cys Asn Leu Leu Glu           
                245                 250                 255               

ggt gag cca agg gag ttt gtg gag aac tct gag tgc ata cag tgc cac        816
Gly Glu Pro Arg Glu Phe Val Glu Asn Ser Glu Cys Ile Gln Cys His           
            260                 265                 270                   

cca gag tgc ctg cct cag gcc atg aac atc acc tgc aca gga cgg gga        864
Pro Glu Cys Leu Pro Gln Ala Met Asn Ile Thr Cys Thr Gly Arg Gly           
        275                 280                 285                       

cca gac aac tgt atc cag tgt gcc cac tac att gac ggc ccc cac tgc        912
Pro Asp Asn Cys Ile Gln Cys Ala His Tyr Ile Asp Gly Pro His Cys           
    290                 295                 300                           

gtc aag acc tgc ccg gca gga gtc atg gga gaa aac aac acc ctg gtc        960
Val Lys Thr Cys Pro Ala Gly Val Met Gly Glu Asn Asn Thr Leu Val           
305                 310                 315                 320           

tgg aag tac gca gac gcc ggc cat gtg tgc cac ctg tgc cat cca aac       1008
Trp Lys Tyr Ala Asp Ala Gly His Val Cys His Leu Cys His Pro Asn           
                325                 330                 335               

tgc acc tac gga tgc act ggg cca ggt ctt gaa ggc tgt cca acg aat       1056
Cys Thr Tyr Gly Cys Thr Gly Pro Gly Leu Glu Gly Cys Pro Thr Asn           
            340                 345                 350                   

ggg cct aag atc ccg tcc atc gcc act ggg atg gtg ggg gcc ctc ctc       1104
Gly Pro Lys Ile Pro Ser Ile Ala Thr Gly Met Val Gly Ala Leu Leu           
        355                 360                 365                       

ttg ctg ctg gtg gtg gcc ctg ggg atc ggc ctc ttc atg tga               1146
Leu Leu Leu Val Val Ala Leu Gly Ile Gly Leu Phe Met                       
    370                 375                 380                           


<210>  15
<211>  381
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic Construct

<400>  15

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


Val Glu Glu Asn Pro Gly Pro Arg Met Leu Leu Leu Val Thr Ser Leu 
            20                  25                  30          


Leu Leu Cys Glu Leu Pro His Pro Ala Phe Leu Leu Ile Pro Arg Lys 
        35                  40                  45              


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


Asn Ala Thr Asn Ile Lys His Phe Lys Asn Cys Thr Ser Ile Ser Gly 
65                  70                  75                  80  


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


Thr Pro Pro Leu Asp Pro Gln Glu Leu Asp Ile Leu Lys Thr Val Lys 
            100                 105                 110         


Glu Ile Thr Gly Phe Leu Leu Ile Gln Ala Trp Pro Glu Asn Arg Thr 
        115                 120                 125             


Asp Leu His Ala Phe Glu Asn Leu Glu Ile Ile Arg Gly Arg Thr Lys 
    130                 135                 140                 


Gln His Gly Gln Phe Ser Leu Ala Val Val Ser Leu Asn Ile Thr Ser 
145                 150                 155                 160 


Leu Gly Leu Arg Ser Leu Lys Glu Ile Ser Asp Gly Asp Val Ile Ile 
                165                 170                 175     


Ser Gly Asn Lys Asn Leu Cys Tyr Ala Asn Thr Ile Asn Trp Lys Lys 
            180                 185                 190         


Leu Phe Gly Thr Ser Gly Gln Lys Thr Lys Ile Ile Ser Asn Arg Gly 
        195                 200                 205             


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


Pro Glu Gly Cys Trp Gly Pro Glu Pro Arg Asp Cys Val Ser Cys Arg 
225                 230                 235                 240 


Asn Val Ser Arg Gly Arg Glu Cys Val Asp Lys Cys Asn Leu Leu Glu 
                245                 250                 255     


Gly Glu Pro Arg Glu Phe Val Glu Asn Ser Glu Cys Ile Gln Cys His 
            260                 265                 270         


Pro Glu Cys Leu Pro Gln Ala Met Asn Ile Thr Cys Thr Gly Arg Gly 
        275                 280                 285             


Pro Asp Asn Cys Ile Gln Cys Ala His Tyr Ile Asp Gly Pro His Cys 
    290                 295                 300                 


Val Lys Thr Cys Pro Ala Gly Val Met Gly Glu Asn Asn Thr Leu Val 
305                 310                 315                 320 


Trp Lys Tyr Ala Asp Ala Gly His Val Cys His Leu Cys His Pro Asn 
                325                 330                 335     


Cys Thr Tyr Gly Cys Thr Gly Pro Gly Leu Glu Gly Cys Pro Thr Asn 
            340                 345                 350         


Gly Pro Lys Ile Pro Ser Ile Ala Thr Gly Met Val Gly Ala Leu Leu 
        355                 360                 365             


Leu Leu Leu Val Val Ala Leu Gly Ile Gly Leu Phe Met 
    370                 375                 380     


