                         SEQUENCE LISTING

<110>  The University of North Carolina at Chapel Hill
       Su, Lishan
 
<120>  CHIMERIC ANTIGEN RECEPTOR CONSTRUCTS AND THEIR USE IN CAR-T CELLS

<130>  5470-846WO

<150>  US 62/839,175
<151>  2019-04-26

<160>  7     

<170>  PatentIn version 3.5

<210>  1
<211>  83
<212>  PRT
<213>  Artificial

<220>
<223>  HVEM CSS domain

<400>  1

Trp Val Trp Trp Phe Leu Ser Gly Ser Leu Val Ile Val Ile Val Cys 
1               5                   10                  15      


Ser Thr Val Gly Leu Ile Ile Cys Val Lys Arg Arg Lys Pro Arg Gly 
            20                  25                  30          


Asp Val Val Lys Val Ile Val Ser Val Gln Arg Lys Arg Gln Glu Ala 
        35                  40                  45              


Glu Gly Glu Ala Thr Val Ile Glu Ala Leu Gln Ala Pro Pro Asp Val 
    50                  55                  60                  


Thr Thr Val Ala Val Glu Glu Thr Ile Pro Ser Phe Thr Gly Arg Ser 
65                  70                  75                  80  


Pro Asn His 
            


<210>  2
<211>  68
<212>  PRT
<213>  Artificial

<220>
<223>  HVEM CSS domain

<400>  2

Leu Val Ile Val Ile Val Cys Ser Thr Val Gly Leu Ile Ile Cys Val 
1               5                   10                  15      


Lys Arg Arg Lys Pro Arg Gly Asp Val Val Lys Val Ile Val Ser Val 
            20                  25                  30          


Gln Arg Lys Arg Gln Glu Ala Glu Gly Glu Ala Thr Val Ile Glu Ala 
        35                  40                  45              


Leu Gln Ala Pro Pro Asp Val Thr Thr Val Ala Val Glu Glu Thr Ile 
    50                  55                  60                  


Pro Ser Phe Thr 
65              


<210>  3
<211>  15
<212>  PRT
<213>  Artificial

<220>
<223>  linker

<400>  3

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 
1               5                   10                  15  


<210>  4
<211>  202
<212>  PRT
<213>  Artificial

<220>
<223>  soluble CD4 domain

<400>  4

Met Asn Arg Gly Val Pro Phe Arg His Leu Leu Leu Val Leu Gln Leu 
1               5                   10                  15      


Ala Leu Leu Pro Ala Ala Thr Gln Gly Lys Lys Val Val Leu Gly Lys 
            20                  25                  30          


Lys Gly Asp Thr Val Glu Leu Thr Cys Thr Ala Ser Gln Lys Lys Ser 
        35                  40                  45              


Ile Gln Phe His Trp Lys Asn Ser Asn Gln Ile Lys Ile Leu Gly Asn 
    50                  55                  60                  


Gln Gly Ser Phe Leu Thr Lys Gly Pro Ser Lys Leu Asn Asp Ala Asp 
65                  70                  75                  80  


Ser Arg Arg Ser Leu Trp Asp Gln Gly Asn Phe Pro Leu Ile Ile Lys 
                85                  90                  95      


Asn Leu Lys Ile Glu Asp Ser Asp Thr Tyr Ile Cys Glu Val Glu Asp 
            100                 105                 110         


Gln Lys Glu Glu Val Gln Leu Leu Val Phe Gly Leu Thr Ala Asn Ser 
        115                 120                 125             


Asp Thr His Leu Leu Gln Gly Gln Ser Leu Thr Leu Thr Leu Glu Ser 
    130                 135                 140                 


Pro Pro Gly Ser Ser Pro Ser Val Gln Cys Arg Ser Pro Arg Gly Lys 
145                 150                 155                 160 


Asn Ile Gln Gly Gly Lys Thr Leu Ser Val Ser Gln Leu Glu Leu Gln 
                165                 170                 175     


Asp Ser Gly Thr Trp Thr Cys Thr Val Leu Gln Asn Gln Lys Lys Val 
            180                 185                 190         


Glu Phe Lys Ile Asp Ile Val Val Leu Ala 
        195                 200         


<210>  5
<211>  75
<212>  PRT
<213>  Artificial

<220>
<223>  CD8 hinge region

<400>  5

Ala Gly Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu Gly Ala Leu Ser 
1               5                   10                  15      


Asn Ser Ile Met Tyr Phe Ser His Phe Val Pro Val Phe Leu Pro Ala 
            20                  25                  30          


Lys Pro Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr 
        35                  40                  45              


Ile Ala Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Ser Arg Pro Ala 
    50                  55                  60                  


Ala Gly Gly Ala Val His Thr Arg Gly Leu Asp 
65                  70                  75  


<210>  6
<211>  473
<212>  PRT
<213>  Artificial

<220>
<223>  chimeric antigen receptor

<400>  6

Met Asn Arg Gly Val Pro Phe Arg His Leu Leu Leu Val Leu Gln Leu 
1               5                   10                  15      


Ala Leu Leu Pro Ala Ala Thr Gln Gly Lys Lys Val Val Leu Gly Lys 
            20                  25                  30          


Lys Gly Asp Thr Val Glu Leu Thr Cys Thr Ala Ser Gln Lys Lys Ser 
        35                  40                  45              


Ile Gln Phe His Trp Lys Asn Ser Asn Gln Ile Lys Ile Leu Gly Asn 
    50                  55                  60                  


Gln Gly Ser Phe Leu Thr Lys Gly Pro Ser Lys Leu Asn Asp Arg Ala 
65                  70                  75                  80  


Asp Ser Arg Arg Ser Leu Trp Asp Gln Gly Asn Phe Pro Leu Ile Ile 
                85                  90                  95      


Lys Asn Leu Lys Ile Glu Asp Ser Asp Thr Tyr Ile Cys Glu Val Glu 
            100                 105                 110         


Asp Gln Lys Glu Glu Val Gln Leu Leu Val Phe Gly Leu Thr Ala Asn 
        115                 120                 125             


Ser Asp Thr His Leu Leu Gln Gly Gln Ser Leu Thr Leu Thr Leu Glu 
    130                 135                 140                 


Ser Pro Pro Gly Ser Ser Pro Ser Val Gln Cys Arg Ser Pro Arg Gly 
145                 150                 155                 160 


Lys Asn Ile Gln Gly Gly Lys Thr Leu Ser Val Ser Gln Leu Glu Leu 
                165                 170                 175     


Gln Asp Ser Gly Thr Trp Thr Cys Thr Val Leu Gln Asn Gln Lys Lys 
            180                 185                 190         


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


Leu Ile Ser Glu Glu Asp Leu Gly Ala Leu Ser Asn Ser Ile Met Tyr 
    210                 215                 220                 


Phe Ser His Phe Val Pro Val Phe Leu Pro Ala Lys Pro Thr Thr Thr 
225                 230                 235                 240 


Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala Ser Gln Pro 
                245                 250                 255     


Leu Ser Leu Arg Pro Glu Ala Ser Arg Pro Ala Ala Gly Gly Ala Val 
            260                 265                 270         


His Thr Arg Gly Leu Asp Trp Val Trp Trp Phe Leu Ser Gly Ser Leu 
        275                 280                 285             


Val Ile Val Ile Val Cys Ser Thr Val Gly Leu Ile Ile Cys Val Lys 
    290                 295                 300                 


Arg Arg Lys Pro Arg Gly Asp Val Val Lys Val Ile Val Ser Val Gln 
305                 310                 315                 320 


Arg Lys Arg Gln Glu Ala Glu Gly Glu Ala Thr Val Ile Glu Ala Leu 
                325                 330                 335     


Gln Ala Pro Pro Asp Val Thr Thr Val Ala Val Glu Glu Thr Ile Pro 
            340                 345                 350         


Ser Phe Thr Gly Arg Ser Pro Asn His Arg Val Lys Phe Ser Arg Ser 
        355                 360                 365             


Ala Asp Ala Pro Ala Tyr Gln Gln Gly Gln Asn Gln Leu Tyr Asn Glu 
    370                 375                 380                 


Leu Asn Leu Gly Arg Arg Glu Glu Tyr Asp Val Leu Asp Lys Arg Arg 
385                 390                 395                 400 


Gly Arg Asp Pro Glu Met Gly Gly Lys Pro Arg Arg Lys Asn Pro Gln 
                405                 410                 415     


Glu Gly Leu Tyr Asn Glu Leu Gln Lys Asp Lys Met Ala Glu Ala Tyr 
            420                 425                 430         


Ser Glu Ile Gly Met Lys Gly Glu Arg Arg Arg Gly Lys Gly His Asp 
        435                 440                 445             


Gly Leu Tyr Gln Gly Leu Ser Thr Ala Thr Lys Asp Thr Tyr Asp Ala 
    450                 455                 460                 


Leu His Met Gln Ala Leu Pro Pro Arg 
465                 470             


<210>  7
<211>  1422
<212>  DNA
<213>  Artificial

<220>
<223>  chimeric antigen receptor

<400>  7
atgaaccggg gagtcccttt taggcacttg cttctggtgc tgcaactggc gctcctccca       60

gcagccactc agggaaagaa agtggtgctg ggcaaaaaag gggatacagt ggaactgacc      120

tgtacagctt cccagaagaa gagcatacaa ttccactgga aaaactccaa ccagataaag      180

attctgggaa atcagggctc cttcttaact aaaggtccat ccaagctgaa tgatcgcgct      240

gactcaagaa gaagcctttg ggaccaagga aacttccccc tgatcatcaa gaatcttaag      300

atagaagact cagatactta catctgtgaa gtggaggacc agaaggagga ggtgcaattg      360

ctagtgttcg gattgactgc caactctgac acccacctgc ttcaggggca gagcctgacc      420

ctgaccttgg agagcccccc tggtagtagc ccctcagtgc aatgtaggag tccaaggggt      480

aaaaacatac agggggggaa gaccctctcc gtgtctcagc tggagctcca ggatagtggc      540

acctggacat gcactgtctt gcagaaccag aagaaggtgg agttcaaaat agacatcgtg      600

gtgctagctg aattcgagca gaagctgatc agcgaggagg acctgggcgc cctgagcaac      660

agcatcatgt acttcagcca cttcgtgccc gtgttcctgc ccgccaagcc caccaccacc      720

cccgcccccc gcccccccac ccccgccccc accatcgcca gccagcccct gagcctgcgc      780

cccgaggcca gccgccccgc cgccggcggc gccgtgcaca cccgcggcct ggactgggtg      840

tggtggttcc tgagcggcag cctggtgatc gtgatcgtgt gcagcaccgt gggcctgatc      900

atctgcgtga agcgccgcaa gccccgcggc gacgtggtga aggtgatcgt gagcgtgcag      960

cgcaagcgcc aggaggccga gggcgaggcc accgtgatcg aggccctgca ggcccccccc     1020

gacgtgacca ccgtggccgt ggaggagacc atccccagct tcaccggccg cagccccaac     1080

caccgcgtga agttcagccg cagcgccgac gcccccgcct accagcaggg ccagaaccag     1140

ctgtacaacg agctgaacct gggccgccgc gaggagtacg acgtgctgga caagcgccgc     1200

ggccgcgacc ccgagatggg cggcaagccc cgccgcaaga acccccagga gggcctgtac     1260

aacgagctgc agaaggacaa gatggccgag gcctacagcg agatcggcat gaagggcgag     1320

cgccgccgcg gcaagggcca cgacggcctg taccagggcc tgagcaccgc caccaaggac     1380

acctacgacg ccctgcacat gcaggccctg cccccccgct aa                        1422


