                         SEQUENCE LISTING

<110>  Beijing Marino Biotechnology Pty Ltd.
 
<120>  Construct, Genetically Modified Lymphocyte, Preparation 
Method And Usage Thereof


<130>  PIDC3154579P

<160>  6     

<170>  PatentIn version 3.3

<210>  1
<211>  255
<212>  PRT
<213>  Homo sapiens

<400>  1

Met Gly Asn Ser Cys Tyr Asn Ile Val Ala Thr Leu Leu Leu Val Leu 
1               5                   10                  15      


Asn Phe Glu Arg Thr Arg Ser Leu Gln Asp Pro Cys Ser Asn Cys Pro 
            20                  25                  30          


Ala Gly Thr Phe Cys Asp Asn Asn Arg Asn Gln Ile Cys Ser Pro Cys 
        35                  40                  45              


Pro Pro Asn Ser Phe Ser Ser Ala Gly Gly Gln Arg Thr Cys Asp Ile 
    50                  55                  60                  


Cys Arg Gln Cys Lys Gly Val Phe Arg Thr Arg Lys Glu Cys Ser Ser 
65                  70                  75                  80  


Thr Ser Asn Ala Glu Cys Asp Cys Thr Pro Gly Phe His Cys Leu Gly 
                85                  90                  95      


Ala Gly Cys Ser Met Cys Glu Gln Asp Cys Lys Gln Gly Gln Glu Leu 
            100                 105                 110         


Thr Lys Lys Gly Cys Lys Asp Cys Cys Phe Gly Thr Phe Asn Asp Gln 
        115                 120                 125             


Lys Arg Gly Ile Cys Arg Pro Trp Thr Asn Cys Ser Leu Asp Gly Lys 
    130                 135                 140                 


Ser Val Leu Val Asn Gly Thr Lys Glu Arg Asp Val Val Cys Gly Pro 
145                 150                 155                 160 


Ser Pro Ala Asp Leu Ser Pro Gly Ala Ser Ser Val Thr Pro Pro Ala 
                165                 170                 175     


Pro Ala Arg Glu Pro Gly His Ser Pro Gln Ile Ile Ser Phe Phe Leu 
            180                 185                 190         


Ala Leu Thr Ser Thr Ala Leu Leu Phe Leu Leu Phe Phe Leu Thr Leu 
        195                 200                 205             


Arg Phe Ser Val Val Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe 
    210                 215                 220                 


Lys Gln Pro Phe Met Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly 
225                 230                 235                 240 


Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu 
                245                 250                 255 


<210>  2
<211>  489
<212>  PRT
<213>  Artificial

<220>
<223>   Amino acid sequence of CD19-specific chimeric antigen receptor

<400>  2

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 Ala Ala Ala Ile Glu 
            260                 265                 270         


Val Met Tyr Pro Pro Pro Tyr Leu Asp Asn Glu Lys Ser Asn Gly Thr 
        275                 280                 285             


Ile Ile His Val Lys Gly Lys His Leu Cys Pro Ser Pro Leu Phe Pro 
    290                 295                 300                 


Gly Pro Ser Lys Pro Phe Trp Val Leu Val Val Val Gly Gly Val Leu 
305                 310                 315                 320 


Ala Cys Tyr Ser Leu Leu Val Thr Val Ala Phe Ile Ile Phe Trp Val 
                325                 330                 335     


Arg Ser Lys Arg Ser Arg Leu Leu His Ser Asp Tyr Met Asn Met Thr 
            340                 345                 350         


Pro Arg Arg Pro Gly Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala Pro 
        355                 360                 365             


Pro Arg Asp Phe Ala Ala Tyr Arg Ser Arg Val Lys Phe Ser Arg Ser 
    370                 375                 380                 


Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu 
385                 390                 395                 400 


Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg 
                405                 410                 415     


Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln 
            420                 425                 430         


Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr 
        435                 440                 445             


Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp 
    450                 455                 460                 


Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala 
465                 470                 475                 480 


Leu His Met Gln Ala Leu Pro Pro Arg 
                485                 


<210>  3
<211>  2899
<212>  DNA
<213>  Artificial

<220>
<223>  nucleic acid sequence of human CD19-targeted CAR-IRES-4-1BB

<400>  3
agatctacta gttctagaat gcttctcctg gtgacaagcc ttctgctctg tgagttacca     60

cacccagcat tcctcctgat cccagacatc cagatgacac agactacatc ctccctgtct    120

gcctctctgg gagacagagt caccatcagt tgcagggcaa gtcaggacat tagtaaatat    180

ttaaattggt atcagcagaa accagatgga actgttaaac tcctgatcta ccatacatca    240

agattacact caggagtccc atcaaggttc agtggcagtg ggtctggaac agattattct    300

ctcaccatta gcaacctgga gcaagaagat attgccactt acttttgcca acagggtaat    360

acgcttccgt acacgttcgg aggggggact aagttggaaa taacaggctc cacctctgga    420

tccggcaagc ccggatctgg cgagggatcc accaagggcg aggtgaaact gcaggagtca    480

ggacctggcc tggtggcgcc ctcacagagc ctgtccgtca catgcactgt ctcaggggtc    540

tcattacccg actatggtgt aagctggatt cgccagcctc cacgaaaggg tctggagtgg    600

ctgggagtaa tatggggtag tgaaaccaca tactataatt cagctctcaa atccagactg    660

accatcatca aggacaactc caagagccaa gttttcttaa aaatgaacag tctgcaaact    720

gatgacacag ccatttacta ctgtgccaaa cattattact acggtggtag ctatgctatg    780

gactactggg gtcaaggaac ctcagtcacc gtctcctcag cggccgcaat tgaagttatg    840

tatcctcctc cttacctaga caatgagaag agcaatggaa ccattatcca tgtgaaaggg    900

aaacaccttt gtccaagtcc cctatttccc ggaccttcta agcccttttg ggtgctggtg    960

gtggttgggg gagtcctggc ttgctatagc ttgctagtaa cagtggcctt tattattttc   1020

tgggtgagga gtaagaggag caggctcctg cacagtgact acatgaacat gactccccgc   1080

cgccccgggc ccacccgcaa gcattaccag ccctatgccc caccacgcga cttcgcagcc   1140

tatcgctcca gagtgaagtt cagcaggagc gcagacgccc ccgcgtacca gcagggccag   1200

aaccagctct ataacgagct caatctagga cgaagagagg agtacgatgt tttggacaag   1260

agacgtggcc gggaccctga gatgggggga aagccgagaa ggaagaaccc tcaggaaggc   1320

ctgtacaatg aactgcagaa agataagatg gcggaggcct acagtgagat tgggatgaaa   1380

ggcgagcgcc ggaggggcaa ggggcacgat ggcctttacc agggtctcag tacagccacc   1440

aaggacacct acgacgccct tcacatgcag gccctgcccc ctcgctaaaa gcttggtacc   1500

ctcgaggtta acgaattccg cccccccccc ctaacgttac tggccgaagc cgcttggaat   1560

aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg   1620

tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc   1680

tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt   1740

cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg   1800

acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac   1860

cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg   1920

tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg   1980

ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc   2040

cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg   2100

gtctagaatg ggaaacagct gttacaacat agtagccact ctgttgctgg tcctcaactt   2160

tgagaggaca agatcattgc aggatccttg tagtaactgc ccagctggta cattctgtga   2220

taataacagg aatcagattt gcagtccctg tcctccaaat agtttctcca gcgcaggtgg   2280

acaaaggacc tgtgacatat gcaggcagtg taaaggtgtt ttcaggacca ggaaggagtg   2340

ttcctccacc agcaatgcag agtgtgactg cactccaggg tttcactgcc tgggggcagg   2400

atgcagcatg tgtgaacagg attgtaaaca aggtcaagaa ctgacaaaaa aaggttgtaa   2460

agactgttgc tttgggacat ttaacgatca gaaacgtggc atctgtcgac cctggacaaa   2520

ctgttctttg gatggaaagt ctgtgcttgt gaatgggacg aaggagaggg acgtggtctg   2580

tggaccatct ccagccgacc tctctccggg agcatcctct gtgaccccgc ctgcccctgc   2640

gagagagcca ggacactctc cgcagatcat ctccttcttt cttgcgctga cgtcgactgc   2700

gttgctcttc ctgctgttct tcctcacgct ccgtttctct gttgttaaac ggggcagaaa   2760

gaaactcctg tatatattca aacaaccatt tatgagacca gtacaaacta ctcaagagga   2820

agatggctgt agctgccgat ttccagaaga agaagaagga ggatgtgaac tgtgaatcct   2880

actgcttcga attaattaa                                                2899


<210>  4
<211>  2964
<212>  DNA
<213>  Artificial

<220>
<223>  nucleic acid of human CD19-targeted CAR-IRES-OX40

<400>  4
agatctacta gttctagaat gcttctcctg gtgacaagcc ttctgctctg tgagttacca     60

cacccagcat tcctcctgat cccagacatc cagatgacac agactacatc ctccctgtct    120

gcctctctgg gagacagagt caccatcagt tgcagggcaa gtcaggacat tagtaaatat    180

ttaaattggt atcagcagaa accagatgga actgttaaac tcctgatcta ccatacatca    240

agattacact caggagtccc atcaaggttc agtggcagtg ggtctggaac agattattct    300

ctcaccatta gcaacctgga gcaagaagat attgccactt acttttgcca acagggtaat    360

acgcttccgt acacgttcgg aggggggact aagttggaaa taacaggctc cacctctgga    420

tccggcaagc ccggatctgg cgagggatcc accaagggcg aggtgaaact gcaggagtca    480

ggacctggcc tggtggcgcc ctcacagagc ctgtccgtca catgcactgt ctcaggggtc    540

tcattacccg actatggtgt aagctggatt cgccagcctc cacgaaaggg tctggagtgg    600

ctgggagtaa tatggggtag tgaaaccaca tactataatt cagctctcaa atccagactg    660

accatcatca aggacaactc caagagccaa gttttcttaa aaatgaacag tctgcaaact    720

gatgacacag ccatttacta ctgtgccaaa cattattact acggtggtag ctatgctatg    780

gactactggg gtcaaggaac ctcagtcacc gtctcctcag cggccgcaat tgaagttatg    840

tatcctcctc cttacctaga caatgagaag agcaatggaa ccattatcca tgtgaaaggg    900

aaacaccttt gtccaagtcc cctatttccc ggaccttcta agcccttttg ggtgctggtg    960

gtggttgggg gagtcctggc ttgctatagc ttgctagtaa cagtggcctt tattattttc   1020

tgggtgagga gtaagaggag caggctcctg cacagtgact acatgaacat gactccccgc   1080

cgccccgggc ccacccgcaa gcattaccag ccctatgccc caccacgcga cttcgcagcc   1140

tatcgctcca gagtgaagtt cagcaggagc gcagacgccc ccgcgtacca gcagggccag   1200

aaccagctct ataacgagct caatctagga cgaagagagg agtacgatgt tttggacaag   1260

agacgtggcc gggaccctga gatgggggga aagccgagaa ggaagaaccc tcaggaaggc   1320

ctgtacaatg aactgcagaa agataagatg gcggaggcct acagtgagat tgggatgaaa   1380

ggcgagcgcc ggaggggcaa ggggcacgat ggcctttacc agggtctcag tacagccacc   1440

aaggacacct acgacgccct tcacatgcag gccctgcccc ctcgctaaaa gcttggtacc   1500

ctcgaggtta acgaattccg cccccccccc ctaacgttac tggccgaagc cgcttggaat   1560

aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg   1620

tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc   1680

tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt   1740

cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg   1800

acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac   1860

cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg   1920

tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg   1980

ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc   2040

cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg   2100

gtctagaatg tgcgtggggg ctcggcggct gggccgcggg ccgtgtgcgg ctctgctcct   2160

cctgggcctg gggctgagca ccgtgacggg gctccactgt gtcggggaca cctaccccag   2220

caacgaccgg tgctgccacg agtgcaggcc aggcaacggg atggtgagcc gctgcagccg   2280

ctcccagaac acggtgtgcc gtccgtgcgg gccgggcttc tacaacgacg tggtcagctc   2340

caagccgtgc aagccctgca cgtggtgtaa cctcagaagt gggagtgagc ggaagcagct   2400

gtgcacggcc acacaggaca cagtctgccg ctgccgggcg ggcacccagc ccctggacag   2460

ctacaagcct ggagttgact gtgccccctg ccctccaggg cacttctccc caggcgacaa   2520

ccaggcctgc aagccctgga ccaactgcac cttggctggg aagcacaccc tgcagccggc   2580

cagcaatagc tcggacgcaa tctgtgagga cagggacccc ccagccacgc agccccagga   2640

gacccagggc cccccggcca ggcccatcac tgtccagccc actgaagcct ggcccagaac   2700

ctcacaggga ccctccaccc ggcccgtgga ggtccccggg ggccgtgcgg ttgccgccat   2760

cctgggcctg ggcctggtgc tggggctgct gggccccctg gccatcctgc tggccctgta   2820

cctgctccgg agggaccaga ggctgccccc cgatgcccac aagccccctg ggggaggcag   2880

tttccggacc cccatccaag aggagcaggc cgacgcccac tccaccctgg ccaagatctg   2940

atcctactgc ttcgaattaa ttaa                                          2964


<210>  5
<211>  589
<212>  DNA
<213>  Artificial

<220>
<223>  nucleic acid sequence of IRES

<400>  5
gttaacgaat tccgcccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc     60

ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg    120

cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca    180

aaggaatgca aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa    240

gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt    300

gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt    360

gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca    420

acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc    480

ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc    540

acggggacgt ggttttcctt tgaaaaacac gatgataata tggccacaa                589


<210>  6
<211>  22
<212>  PRT
<213>  Artificial

<220>
<223>   Amino acid sequence of the cleavable linker peptide

<400>  6

Gly Ser Gly Ala Thr Asn Phe Ser Leu Leu Lys Gln Ala Gly Asp Val 
1               5                   10                  15      


Glu Glu Asn Pro Gly Pro 
            20          


