               SEQUENCE LISTING

<110> PRINCE OF SONGKLA UNIVERSITY 

<120> CD28/CD40 CO-STIMULATORY DOMAIN OF THE CHIMERIC ANTIGEN RECEPTOR

<140> THPCT/2022/000009
<141> 2022-03-21

<160> 18

<170> BiSSAP 1.3.6

<210> 1
<211> 123
<212> DNA
<213> Artificial Sequence


<220> CD19.28.40z CAR gene sequence
<223> CD28 co-stimulatory domain

<400> 1
aggagtaaga ggagcaggct cctgcacagt gactacatga acatgactcc ccgccgcccc     60

gggcccaccc gcaagcatta ccagccctat gccccaccac gcgacttcgc agcctatcgc    120

tcc                                                                  123


<210> 2
<211> 9
<212> DNA
<213> Artificial Sequence


<220> CD19.28.40z CAR gene sequence
<223> some nucleotide sequence to link between CD28 and CD40

<400> 2
ggcggaggg                                                           9


<210> 3
<211> 186
<212> DNA
<213> Artificial Sequence


<220> CD19.28.40z CAR gene sequence
<223> CD40 co-stimulatory domain

<400> 3
aaaaaggtgg ccaagaagcc aaccaataag gccccccacc ccaagcagga accccaggag     60

atcaattttc ccgacgatct tcctggctcc aacactgctg ctccagtgca ggagacttta    120

catggatgcc aaccggtcac ccaggaggat ggcaaagaga gtcgcatctc agtgcaggag    180

agacag                                                               186


<210> 4
<211> 106
<212> PRT
<213> Artificial Sequence


<220> CD19.28.40z CAR gene sequence
<223> The translated amino acid sequence of CD28/CD40 protein

<400> 4
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 Gly Gly Gly Lys Lys Val Ala 
        35                  40                  45              
Lys Lys Pro Thr Asn Lys Ala Pro His Pro Lys Gln Glu Pro Gln Glu 
    50                  55                  60                  
Ile Asn Phe Pro Asp Asp Leu Pro Gly Ser Asn Thr Ala Ala Pro Val 
65                  70                  75                  80  
Gln Glu Thr Leu His Gly Cys Gln Pro Val Thr Gln Glu Asp Gly Lys 
                85                  90                  95      
Glu Ser Arg Ile Ser Val Gln Glu Arg Gln 
            100                 105     

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


<220> Primer for fragment 1 DNA synthesis: HindIII-CD19scFV-28TM-CD28-CD40
<223> Forward primer HindIII-19CARscFv (5'-3')

<400> 5
aagcttatgc tgctgctggt gaccagcctg ctg                                  33


<210> 6
<211> 33
<212> DNA
<213> Artificial Sequence


<220> Primer for fragment 1 DNA synthesis: HindIII-CD19scFV-28TM-CD28-CD40
<223> Reverse primer CD40-GGG-CD28IC (5'-3')

<400> 6
ctttttccct ccgccggagc gataggctgc gaa                                  33


<210> 7
<211> 33
<212> DNA
<213> Artificial Sequence


<220> Primer for Fragment 2 DNA synthesis: CD28-CD40-CD3z
<223> Forward primer CD28IC-GGG-CD40 (5'-3')

<400> 7
gcagcctatc gctccggcgg agggaaaaag gtg                                  33


<210> 8
<211> 33
<212> DNA
<213> Artificial Sequence


<220> Primer for Fragment 2 DNA synthesis: CD28-CD40-CD3z
<223> Reverse primer CD3z-CD40 (5'-3')

<400> 8
gctgaacttc acccgctgtc tctcctgcac tga                                  33


<210> 9
<211> 33
<212> DNA
<213> Artificial Sequence


<220> Primer for Fragment 3 DNA synthesis: CD40-CD3z-tEGFR-NotI 
<223> Forward primer CD40-CD3z (5'-3')

<400> 9
gtgcaggaga gacagcgggt gaagttcagc aga                                  33


<210> 10
<211> 33
<212> DNA
<213> Artificial Sequence


<220> Primer for Fragment 3 DNA synthesis: CD40-CD3z-tEGFR-NotI 
<223> Reverse primer NotI-tEGFR (5'-3')

<400> 10
gcggccgctc acatgaagag gccgatcccc agg                                  33


<210> 11
<211> 33
<212> DNA
<213> Artificial Sequence


<220> Primer to confirm the completion of the product of CD19.28.40z from 3 fragments 
<223> Forward primer HindIII-19CARscFv (5'-3')

<400> 11
aagcttatgc tgctgctggt gaccagcctg ctg                                  33


<210> 12
<211> 33
<212> DNA
<213> Artificial Sequence


<220> Primer to confirm the completion of the product of CD19.28.40z from 3 fragments
<223> Reverse primer NotI-tEGFR (5'-3')

<400> 12
gcggccgctc acatgaagag gccgatcccc agg                                  33


<210> 13
<211> 33
<212> DNA
<213> Artificial Sequence


<220> Primer to confirm the completion check of plasmid containing CD19.28.40z CAR 
<223> Forward primer LZRS-HindIII (5'-3')

<400> 13
gatcccagtg tggtggtagg gaattcaaag ctt                                  33


<210> 14
<211> 33
<212> DNA
<213> Artificial Sequence


<220> Primer to confirm the completion check of plasmid containing CD19.28.40z CAR 
<223> Reverse primer LZRS-NotI (5'-3')

<400> 14
tattttatcg tcgaccactg tgctggcggc cgc                                  33


<210> 15
<211> 33
<212> DNA
<213> Artificial Sequence


<220> Primers used in CD19.28.40z CAR sequencing
<223> Forward primer LZRS-HindIII (5'-3')

<400> 15
gatcccagtg tggtggtagg gaattcaaag ctt                                  33


<210> 16
<211> 19
<212> DNA
<213> Artificial Sequence


<220> Primers used in CD19.28.40z CAR sequencing
<223> Forward primer CD28TM (5'-3')

<400> 16
atgttctggg tgctggtgg                                                 19


<210> 17
<211> 20
<212> DNA
<213> Artificial Sequence


<220> Primers used in CD19.28.40z CAR sequencing
<223> Forward primer CD3z-ITAM2 (5'-3')

<400> 17
ccaggaaggc ctgtataacg                                                20


<210> 18
<211> 24
<212> DNA
<213> Artificial Sequence


<220> Primers used in CD19.28.40z CAR sequencing
<223> Reverse primer tEGFR (5'-3')

<400> 18
tgatggagat gtgataattt cagg                                           24


