                         SEQUENCE LISTING

<110>  A2 Biotherapeutics, Inc.
       KAMB, Carl Alexander
       OH, Julyun
       WARSHAVIAK, Dora Toledo
       MKRTICHYAN, Mika
       MUNGUIA, Melanie
 
<120>  Bifunctional Single Variable Domain T Cell Receptors and Uses 
       Thereof

<130>  A2BI-010/01WO

<150>  62/885,045
<151>  2019-08-09

<160>  34    

<170>  PatentIn version 3.5

<210>  1
<211>  13
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  NY-ESO CDR3

<400>  1

Cys Ala Ser Ser Leu Gly Leu Gly Tyr Glu Gln Tyr Phe 
1               5                   10              


<210>  2
<211>  20
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  NY-ESO CDR3

<400>  2

Cys Ala Ser Ser Leu Gly Gly Pro Arg Gly Leu Ala Gly Leu Arg Gly 
1               5                   10                  15      


Asp Glu Gln Phe 
            20  


<210>  3
<211>  12
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  NY-ESO CDR3

<400>  3

Cys Ala Ser Ser Leu Arg Arg Asp Asn Glu Gln Phe 
1               5                   10          


<210>  4
<211>  19
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  MAGE-A3 CDR3

<400>  4

Cys Ala Ser Ser Leu Glu Val Leu Leu Gly Ala Asp Phe Pro Asp Thr 
1               5                   10                  15      


Gln Tyr Phe 
            


<210>  5
<211>  18
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  MAGE-A3 CDR3

<400>  5

Cys Ala Ser Ser Phe Pro Ala Gly His Gly Ala Asp Leu Asp Asn Glu 
1               5                   10                  15      


Gln Phe 
        


<210>  6
<211>  14
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  MAGE-A3 CDR3

<400>  6

Cys Ala Ser Ser Glu Ile Thr Gly Arg Ile Gly Glu Gln Phe 
1               5                   10                  


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

<220>
<223>  MAGE-A3 CDR3

<400>  7

Cys Ala Ser Ser Leu Gly Gly Asp Glu Leu Gly Ala Asp Gly Asn Glu 
1               5                   10                  15      


Gln Phe 
        


<210>  8
<211>  45
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  CD8 alpha hinge

<400>  8

Thr Thr Thr Pro Ala Pro Arg Pro Pro Thr Pro Ala Pro Thr Ile Ala 
1               5                   10                  15      


Ser Gln Pro Leu Ser Leu Arg Pro Glu Ala Cys Arg Pro Ala Ala Gly 
            20                  25                  30          


Gly Ala Val His Thr Arg Gly Leu Asp Phe Ala Cys Asp 
        35                  40                  45  


<210>  9
<211>  135
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  CD8 alpha hinge

<400>  9
accacgacgc cagcgccgcg accaccaaca ccggcgccca ccatcgcgtc gcagcccctg       60

tccctgcgcc cagaggcgtg ccggccagcg gcggggggcg cagtgcacac gagggggctg      120

gacttcgcct gtgat                                                       135


<210>  10
<211>  41
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  CD28 hinge

<400>  10

Cys Thr Ile Glu Val Met Tyr Pro Pro Pro Tyr Leu Asp Asn Glu Lys 
1               5                   10                  15      


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


Pro Leu Phe Pro Gly Pro Ser Lys Pro 
        35                  40      


<210>  11
<211>  123
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  CD28 hinge

<400>  11
tgtaccattg aagttatgta tcctcctcct tacctagaca atgagaagag caatggaacc       60

attatccatg tgaaagggaa acacctttgt ccaagtcccc tatttcccgg accttctaag      120

ccc                                                                    123


<210>  12
<211>  23
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  TCR alpha transmembrane domain

<400>  12

Val Ile Gly Phe Arg Ile Leu Leu Leu Lys Val Ala Gly Phe Asn Leu 
1               5                   10                  15      


Leu Met Thr Leu Arg Leu Trp 
            20              


<210>  13
<211>  69
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  TCR alpha transmembrane domain

<400>  13
gtgattgggt tccgaatcct cctcctgaaa gtggccgggt ttaatctgct catgacgctg       60

cggctgtgg                                                               69


<210>  14
<211>  23
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  TCR beta transmembrane domain

<400>  14

Thr Ile Leu Tyr Glu Ile Leu Leu Gly Lys Ala Thr Leu Tyr Ala Val 
1               5                   10                  15      


Leu Val Ser Ala Leu Val Leu 
            20              


<210>  15
<211>  69
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  TCR beta transmembrane domain

<400>  15
accatcctct atgagatctt gctagggaag gccaccttgt atgccgtgct ggtcagtgcc       60

ctcgtgctg                                                               69


<210>  16
<211>  21
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  CD3 zeta transmembrane domain

<400>  16

Leu Cys Tyr Leu Leu Asp Gly Ile Leu Phe Ile Tyr Gly Val Ile Leu 
1               5                   10                  15      


Thr Ala Leu Phe Leu 
            20      


<210>  17
<211>  89
<212>  PRT
<213>  CD3 delta intracellular domain

<400>  17

Gly His Glu Thr Gly Arg Leu Ser Gly Ala Ala Asp Thr Gln Ala Leu 
1               5                   10                  15      


Leu Arg Asn Asp Gln Val Tyr Gln Pro Leu Arg Asp Arg Asp Asp Ala 
            20                  25                  30          


Gln Tyr Ser His Leu Gly Gly Asn Trp Ala Arg Asn Lys Gly Gly Ser 
        35                  40                  45              


Arg Ser Lys Arg Ser Arg Leu Leu His Ser Asp Tyr Met Asn Met Thr 
    50                  55                  60                  


Pro Arg Arg Pro Gly Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala Pro 
65                  70                  75                  80  


Pro Arg Asp Phe Ala Ala Tyr Arg Ser 
                85                  


<210>  18
<211>  269
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  CD3 delta intracelllular domain

<400>  18
ggacatgaga ctggaaggct gtctggggct gccgacacac aagctctgtt gaggaatgac       60

caggtctatc agcccctccg agatcgagat gatgctcagt acagccacct tggaggaaac      120

tgggctcgga acaagggcgg aagcaggagc aagcggagca gactgctgca cagcgactac      180

atgaacatga ccccccggag gcctggcccc acccggaagc actaccagcc ctacgcccct      240

cccagggatt tcgccgccta ccggagcta                                        269


<210>  19
<211>  99
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  CD3 epsilon intracellular domain

<400>  19

Lys Asn Arg Lys Ala Lys Ala Lys Pro Val Thr Arg Gly Ala Gly Ala 
1               5                   10                  15      


Gly Gly Arg Gln Arg Gly Gln Asn Lys Glu Arg Pro Pro Pro Val Pro 
            20                  25                  30          


Asn Pro Asp Tyr Glu Pro Ile Arg Lys Gly Gln Arg Asp Leu Tyr Ser 
        35                  40                  45              


Gly Leu Asn Gln Arg Arg Ile Gly Gly Ser Arg Ser Lys Arg Ser Arg 
    50                  55                  60                  


Leu Leu His Ser Asp Tyr Met Asn Met Thr Pro Arg Arg Pro Gly Pro 
65                  70                  75                  80  


Thr Arg Lys His Tyr Gln Pro Tyr Ala Pro Pro Arg Asp Phe Ala Ala 
                85                  90                  95      


Tyr Arg Ser 
            


<210>  20
<211>  300
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  CD3 epsilon intracellular domain

<400>  20
aagaatagaa aggccaaggc caagcctgtg acacgaggag cgggtgctgg cggcaggcaa       60

aggggacaaa acaaggagag gccaccacct gttcccaacc cagactatga gcccatccgg      120

aaaggccagc gggacctgta ttctggcctg aatcagcgca gaatcggcgg aagcaggagc      180

aagcggagca gactgctgca cagcgactac atgaacatga ccccccggag gcctggcccc      240

acccggaagc actaccagcc ctacgcccct cccagggatt tcgccgccta ccggagctag      300


<210>  21
<211>  89
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  CD3 gamma intracellular domain

<400>  21

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


Leu Pro Asn Asp Gln Leu Tyr Gln Pro Leu Lys Asp Arg Glu Asp Asp 
            20                  25                  30          


Gln Tyr Ser His Leu Gln Gly Asn Gln Leu Arg Arg Asn Gly Gly Ser 
        35                  40                  45              


Arg Ser Lys Arg Ser Arg Leu Leu His Ser Asp Tyr Met Asn Met Thr 
    50                  55                  60                  


Pro Arg Arg Pro Gly Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala Pro 
65                  70                  75                  80  


Pro Arg Asp Phe Ala Ala Tyr Arg Ser 
                85                  


<210>  22
<211>  270
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  CD3 gamma intracellular domain

<400>  22
ggacaggatg gagttcgcca gtcgagagct tcagacaagc agactctgtt gcccaatgac       60

cagctctacc agcccctcaa ggatcgagaa gatgaccagt acagccacct tcaaggaaac      120

cagttgagga ggaatggcgg aagcaggagc aagcggagca gactgctgca cagcgactac      180

atgaacatga ccccccggag gcctggcccc acccggaagc actaccagcc ctacgcccct      240

cccagggatt tcgccgccta ccggagctag                                       270


<210>  23
<211>  42
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  CD3 zeta intracellular domain

<400>  23

Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met 
1               5                   10                  15      


Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe 
            20                  25                  30          


Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu 
        35                  40          


<210>  24
<211>  10
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  TCR beta intracellular domain

<400>  24

Met Ala Met Val Lys Arg Lys Asp Ser Arg 
1               5                   10  


<210>  25
<211>  30
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  TCR beta intracellular domain

<400>  25
atggccatgg tcaagagaaa ggattccaga                                        30


<210>  26
<211>  41
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  CD28 intracellular domain

<400>  26

Arg Ser Lys Arg Ser Arg Leu Leu His Ser Asp Tyr Met Asn Met Thr 
1               5                   10                  15      


Pro Arg Arg Pro Gly Pro Thr Arg Lys His Tyr Gln Pro Tyr Ala Pro 
            20                  25                  30          


Pro Arg Asp Phe Ala Ala Tyr Arg Ser 
        35                  40      


<210>  27
<211>  123
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  CD28 intracellular domain

<400>  27
aggagcaagc ggagcagact gctgcacagc gactacatga acatgacccc ccggaggcct       60

ggccccaccc ggaagcacta ccagccctac gcccctccca gggatttcgc cgcctaccgg      120

agc                                                                    123


<210>  28
<211>  42
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  4-1BB intracellular domain

<400>  28

Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe Lys Gln Pro Phe Met 
1               5                   10                  15      


Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly Cys Ser Cys Arg Phe 
            20                  25                  30          


Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu 
        35                  40          


<210>  29
<211>  126
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  4-1BB intracellular domain

<400>  29
aaacggggca gaaagaaact cctgtatata ttcaaacaac catttatgag gccagtacaa       60

actactcaag aggaagatgg ctgtagctgc cgatttccag aagaagaaga aggaggatgt      120

gaactg                                                                 126


<210>  30
<211>  114
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  TRBV5-8 Vbeta

<400>  30

Met Gly Pro Arg Leu Leu Phe Trp Ala Leu Leu Cys Leu Leu Gly Thr 
1               5                   10                  15      


Gly Pro Val Glu Ala Gly Val Thr Gln Ser Pro Thr His Leu Ile Lys 
            20                  25                  30          


Thr Arg Gly Gln Gln Ala Thr Leu Arg Cys Ser Pro Ile Ser Gly His 
        35                  40                  45              


Thr Ser Val Tyr Trp Tyr Gln Gln Ala Leu Gly Leu Gly Leu Gln Phe 
    50                  55                  60                  


Leu Leu Trp Tyr Asp Glu Gly Glu Glu Arg Asn Arg Gly Asn Phe Pro 
65                  70                  75                  80  


Pro Arg Phe Ser Gly Arg Gln Phe Pro Asn Tyr Ser Ser Glu Leu Asn 
                85                  90                  95      


Val Asn Ala Leu Glu Leu Glu Asp Ser Ala Leu Tyr Leu Cys Ala Ser 
            100                 105                 110         


Ser Leu 
        


<210>  31
<211>  7
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  linkger

<400>  31

Gly Gly Gly Gly Ser Gly Gly 
1               5           


<210>  32
<211>  4
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  linker

<400>  32

Gly Gly Gly Ser 
1               


<210>  33
<211>  5
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  linker

<400>  33

Gly Gly Gly Gly Ser 
1               5   


<210>  34
<211>  9
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  peptide

<400>  34

Ser Leu Leu Met Trp Ile Thr Gln Cys 
1               5                   


