                         SEQUENCE LISTING

<110>  Autolus Limited
 
<120>  Chimeric protein

<130>  P117641PCT

<150>  GB 1906126.6
<151>  2019-05-01

<160>  13    

<170>  PatentIn version 3.5

<210>  1
<211>  404
<212>  PRT
<213>  Homo sapiens

<400>  1

Met Ala Asp Lys Val Leu Lys Glu Lys Arg Lys Leu Phe Ile Arg Ser 
1               5                   10                  15      


Met Gly Glu Gly Thr Ile Asn Gly Leu Leu Asp Glu Leu Leu Gln Thr 
            20                  25                  30          


Arg Val Leu Asn Lys Glu Glu Met Glu Lys Val Lys Arg Glu Asn Ala 
        35                  40                  45              


Thr Val Met Asp Lys Thr Arg Ala Leu Ile Asp Ser Val Ile Pro Lys 
    50                  55                  60                  


Gly Ala Gln Ala Cys Gln Ile Cys Ile Thr Tyr Ile Cys Glu Glu Asp 
65                  70                  75                  80  


Ser Tyr Leu Ala Gly Thr Leu Gly Leu Ser Ala Asp Gln Thr Ser Gly 
                85                  90                  95      


Asn Tyr Leu Asn Met Gln Asp Ser Gln Gly Val Leu Ser Ser Phe Pro 
            100                 105                 110         


Ala Pro Gln Ala Val Gln Asp Asn Pro Ala Met Pro Thr Ser Ser Gly 
        115                 120                 125             


Ser Glu Gly Asn Val Lys Leu Cys Ser Leu Glu Glu Ala Gln Arg Ile 
    130                 135                 140                 


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


Arg Thr Arg Leu Ala Leu Ile Ile Cys Asn Glu Glu Phe Asp Ser Ile 
                165                 170                 175     


Pro Arg Arg Thr Gly Ala Glu Val Asp Ile Thr Gly Met Thr Met Leu 
            180                 185                 190         


Leu Gln Asn Leu Gly Tyr Ser Val Asp Val Lys Lys Asn Leu Thr Ala 
        195                 200                 205             


Ser Asp Met Thr Thr Glu Leu Glu Ala Phe Ala His Arg Pro Glu His 
    210                 215                 220                 


Lys Thr Ser Asp Ser Thr Phe Leu Val Phe Met Ser His Gly Ile Arg 
225                 230                 235                 240 


Glu Gly Ile Cys Gly Lys Lys His Ser Glu Gln Val Pro Asp Ile Leu 
                245                 250                 255     


Gln Leu Asn Ala Ile Phe Asn Met Leu Asn Thr Lys Asn Cys Pro Ser 
            260                 265                 270         


Leu Lys Asp Lys Pro Lys Val Ile Ile Ile Gln Ala Cys Arg Gly Asp 
        275                 280                 285             


Ser Pro Gly Val Val Trp Phe Lys Asp Ser Val Gly Val Ser Gly Asn 
    290                 295                 300                 


Leu Ser Leu Pro Thr Thr Glu Glu Phe Glu Asp Asp Ala Ile Lys Lys 
305                 310                 315                 320 


Ala His Ile Glu Lys Asp Phe Ile Ala Phe Cys Ser Ser Thr Pro Asp 
                325                 330                 335     


Asn Val Ser Trp Arg His Pro Thr Met Gly Ser Val Phe Ile Gly Arg 
            340                 345                 350         


Leu Ile Glu His Met Gln Glu Tyr Ala Cys Ser Cys Asp Val Glu Glu 
        355                 360                 365             


Ile Phe Arg Lys Val Arg Phe Ser Phe Glu Gln Pro Asp Gly Arg Ala 
    370                 375                 380                 


Gln Met Pro Thr Thr Glu Arg Val Thr Leu Thr Arg Cys Phe Tyr Leu 
385                 390                 395                 400 


Phe Pro Gly His 
                


<210>  2
<211>  285
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  truncated human Caspase-1

<400>  2

Asn Pro Ala Met Pro Thr Ser Ser Gly Ser Glu Gly Asn Val Lys Leu 
1               5                   10                  15      


Cys Ser Leu Glu Glu Ala Gln Arg Ile Trp Lys Gln Lys Ser Ala Glu 
            20                  25                  30          


Ile Tyr Pro Ile Met Asp Lys Ser Ser Arg Thr Arg Leu Ala Leu Ile 
        35                  40                  45              


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


Val Asp Ile Thr Gly Met Thr Met Leu Leu Gln Asn Leu Gly Tyr Ser 
65                  70                  75                  80  


Val Asp Val Lys Lys Asn Leu Thr Ala Ser Asp Met Thr Thr Glu Leu 
                85                  90                  95      


Glu Ala Phe Ala His Arg Pro Glu His Lys Thr Ser Asp Ser Thr Phe 
            100                 105                 110         


Leu Val Phe Met Ser His Gly Ile Arg Glu Gly Ile Cys Gly Lys Lys 
        115                 120                 125             


His Ser Glu Gln Val Pro Asp Ile Leu Gln Leu Asn Ala Ile Phe Asn 
    130                 135                 140                 


Met Leu Asn Thr Lys Asn Cys Pro Ser Leu Lys Asp Lys Pro Lys Val 
145                 150                 155                 160 


Ile Ile Ile Gln Ala Cys Arg Gly Asp Ser Pro Gly Val Val Trp Phe 
                165                 170                 175     


Lys Asp Ser Val Gly Val Ser Gly Asn Leu Ser Leu Pro Thr Thr Glu 
            180                 185                 190         


Glu Phe Glu Asp Asp Ala Ile Lys Lys Ala His Ile Glu Lys Asp Phe 
        195                 200                 205             


Ile Ala Phe Cys Ser Ser Thr Pro Asp Asn Val Ser Trp Arg His Pro 
    210                 215                 220                 


Thr Met Gly Ser Val Phe Ile Gly Arg Leu Ile Glu His Met Gln Glu 
225                 230                 235                 240 


Tyr Ala Cys Ser Cys Asp Val Glu Glu Ile Phe Arg Lys Val Arg Phe 
                245                 250                 255     


Ser Phe Glu Gln Pro Asp Gly Arg Ala Gln Met Pro Thr Thr Glu Arg 
            260                 265                 270         


Val Thr Leu Thr Arg Cys Phe Tyr Leu Phe Pro Gly His 
        275                 280                 285 


<210>  3
<211>  516
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  chimeric protein sequence (FRB-L1-FKBP12-L2-dCasp1)

<400>  3

Met Ala Ser Arg Ile Leu Trp His Glu Met Trp His Glu Gly Leu Glu 
1               5                   10                  15      


Glu Ala Ser Arg Leu Tyr Phe Gly Glu Arg Asn Val Lys Gly Met Phe 
            20                  25                  30          


Glu Val Leu Glu Pro Leu His Ala Met Met Glu Arg Gly Pro Gln Thr 
        35                  40                  45              


Leu Lys Glu Thr Ser Phe Asn Gln Ala Tyr Gly Arg Asp Leu Met Glu 
    50                  55                  60                  


Ala Gln Glu Trp Cys Arg Lys Tyr Met Lys Ser Gly Asn Val Lys Asp 
65                  70                  75                  80  


Leu Leu Gln Ala Trp Asp Leu Tyr Tyr His Val Phe Arg Arg Ile Ser 
                85                  90                  95      


Lys Leu Glu Tyr Ser Gly Gly Gly Ser Leu Glu Gly Val Gln Val Glu 
            100                 105                 110         


Thr Ile Ser Pro Gly Asp Gly Arg Thr Phe Pro Lys Arg Gly Gln Thr 
        115                 120                 125             


Cys Val Val His Tyr Thr Gly Met Leu Glu Asp Gly Lys Lys Phe Asp 
    130                 135                 140                 


Ser Ser Arg Asp Arg Asn Lys Pro Phe Lys Phe Met Leu Gly Lys Gln 
145                 150                 155                 160 


Glu Val Ile Arg Gly Trp Glu Glu Gly Val Ala Gln Met Ser Val Gly 
                165                 170                 175     


Gln Arg Ala Lys Leu Thr Ile Ser Pro Asp Tyr Ala Tyr Gly Ala Thr 
            180                 185                 190         


Gly His Pro Gly Ile Ile Pro Pro His Ala Thr Leu Val Phe Asp Val 
        195                 200                 205             


Glu Leu Leu Lys Leu Glu Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 
    210                 215                 220                 


Ser Gly Gly Gly Gly Ser Gly Asn Pro Ala Met Pro Thr Ser Ser Gly 
225                 230                 235                 240 


Ser Glu Gly Asn Val Lys Leu Cys Ser Leu Glu Glu Ala Gln Arg Ile 
                245                 250                 255     


Trp Lys Gln Lys Ser Ala Glu Ile Tyr Pro Ile Met Asp Lys Ser Ser 
            260                 265                 270         


Arg Thr Arg Leu Ala Leu Ile Ile Cys Asn Glu Glu Phe Asp Ser Ile 
        275                 280                 285             


Pro Arg Arg Thr Gly Ala Glu Val Asp Ile Thr Gly Met Thr Met Leu 
    290                 295                 300                 


Leu Gln Asn Leu Gly Tyr Ser Val Asp Val Lys Lys Asn Leu Thr Ala 
305                 310                 315                 320 


Ser Asp Met Thr Thr Glu Leu Glu Ala Phe Ala His Arg Pro Glu His 
                325                 330                 335     


Lys Thr Ser Asp Ser Thr Phe Leu Val Phe Met Ser His Gly Ile Arg 
            340                 345                 350         


Glu Gly Ile Cys Gly Lys Lys His Ser Glu Gln Val Pro Asp Ile Leu 
        355                 360                 365             


Gln Leu Asn Ala Ile Phe Asn Met Leu Asn Thr Lys Asn Cys Pro Ser 
    370                 375                 380                 


Leu Lys Asp Lys Pro Lys Val Ile Ile Ile Gln Ala Cys Arg Gly Asp 
385                 390                 395                 400 


Ser Pro Gly Val Val Trp Phe Lys Asp Ser Val Gly Val Ser Gly Asn 
                405                 410                 415     


Leu Ser Leu Pro Thr Thr Glu Glu Phe Glu Asp Asp Ala Ile Lys Lys 
            420                 425                 430         


Ala His Ile Glu Lys Asp Phe Ile Ala Phe Cys Ser Ser Thr Pro Asp 
        435                 440                 445             


Asn Val Ser Trp Arg His Pro Thr Met Gly Ser Val Phe Ile Gly Arg 
    450                 455                 460                 


Leu Ile Glu His Met Gln Glu Tyr Ala Cys Ser Cys Asp Val Glu Glu 
465                 470                 475                 480 


Ile Phe Arg Lys Val Arg Phe Ser Phe Glu Gln Pro Asp Gly Arg Ala 
                485                 490                 495     


Gln Met Pro Thr Thr Glu Arg Val Thr Leu Thr Arg Cys Phe Tyr Leu 
            500                 505                 510         


Phe Pro Gly His 
        515     


<210>  4
<211>  108
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  dimerization domain (FKBP12 with an F36V mutation)

<400>  4

Met Gly Val Gln Val Glu Thr Ile Ser Pro Gly Asp Gly Arg Thr Phe 
1               5                   10                  15      


Pro Lys Arg Gly Gln Thr Cys Val Val His Tyr Thr Gly Met Leu Glu 
            20                  25                  30          


Asp Gly Lys Lys Val Asp Ser Ser Arg Asp Arg Asn Lys Pro Phe Lys 
        35                  40                  45              


Phe Met Leu Gly Lys Gln Glu Val Ile Arg Gly Trp Glu Glu Gly Val 
    50                  55                  60                  


Ala Gln Met Ser Val Gly Gln Arg Ala Lys Leu Thr Ile Ser Pro Asp 
65                  70                  75                  80  


Tyr Ala Tyr Gly Ala Thr Gly His Pro Gly Ile Ile Pro Pro His Ala 
                85                  90                  95      


Thr Leu Val Phe Asp Val Glu Leu Leu Lys Leu Glu 
            100                 105             


<210>  5
<211>  109
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  chimeric protein heterodimerization domain, FKBP12 domain

<400>  5

Leu Glu Gly Val Gln Val Glu Thr Ile Ser Pro Gly Asp Gly Arg Thr 
1               5                   10                  15      


Phe Pro Lys Arg Gly Gln Thr Cys Val Val His Tyr Thr Gly Met Leu 
            20                  25                  30          


Glu Asp Gly Lys Lys Phe Asp Ser Ser Arg Asp Arg Asn Lys Pro Phe 
        35                  40                  45              


Lys Phe Met Leu Gly Lys Gln Glu Val Ile Arg Gly Trp Glu Glu Gly 
    50                  55                  60                  


Val Ala Gln Met Ser Val Gly Gln Arg Ala Lys Leu Thr Ile Ser Pro 
65                  70                  75                  80  


Asp Tyr Ala Tyr Gly Ala Thr Gly His Pro Gly Ile Ile Pro Pro His 
                85                  90                  95      


Ala Thr Leu Val Phe Asp Val Glu Leu Leu Lys Leu Glu 
            100                 105                 


<210>  6
<211>  99
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  chimeric protein heterodimerization domain, wild-type FRB segment
       of mTOR

<400>  6

Met Ala Ser Arg Ile Leu Trp His Glu Met Trp His Glu Gly Leu Glu 
1               5                   10                  15      


Glu Ala Ser Arg Leu Tyr Phe Gly Glu Arg Asn Val Lys Gly Met Phe 
            20                  25                  30          


Glu Val Leu Glu Pro Leu His Ala Met Met Glu Arg Gly Pro Gln Thr 
        35                  40                  45              


Leu Lys Glu Thr Ser Phe Asn Gln Ala Tyr Gly Arg Asp Leu Met Glu 
    50                  55                  60                  


Ala Gln Glu Trp Cys Arg Lys Tyr Met Lys Ser Gly Asn Val Lys Asp 
65                  70                  75                  80  


Leu Thr Gln Ala Trp Asp Leu Tyr Tyr His Val Phe Arg Arg Ile Ser 
                85                  90                  95      


Lys Leu Glu 
            


<210>  7
<211>  99
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  chimeric protein heterodimerization domain, FRB with T to L 
       substitution at 2098 which allows binding to AP21967

<400>  7

Met Ala Ser Arg Ile Leu Trp His Glu Met Trp His Glu Gly Leu Glu 
1               5                   10                  15      


Glu Ala Ser Arg Leu Tyr Phe Gly Glu Arg Asn Val Lys Gly Met Phe 
            20                  25                  30          


Glu Val Leu Glu Pro Leu His Ala Met Met Glu Arg Gly Pro Gln Thr 
        35                  40                  45              


Leu Lys Glu Thr Ser Phe Asn Gln Ala Tyr Gly Arg Asp Leu Met Glu 
    50                  55                  60                  


Ala Gln Glu Trp Cys Arg Lys Tyr Met Lys Ser Gly Asn Val Lys Asp 
65                  70                  75                  80  


Leu Leu Gln Ala Trp Asp Leu Tyr Tyr His Val Phe Arg Arg Ile Ser 
                85                  90                  95      


Lys Leu Glu 
            


<210>  8
<211>  99
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  chimeric protein heterodimerization domain, FRB segment of mTOR 
       with T to H substitution at 2098 and to W at F at residue 2101

<400>  8

Met Ala Ser Arg Ile Leu Trp His Glu Met Trp His Glu Gly Leu Glu 
1               5                   10                  15      


Glu Ala Ser Arg Leu Tyr Phe Gly Glu Arg Asn Val Lys Gly Met Phe 
            20                  25                  30          


Glu Val Leu Glu Pro Leu His Ala Met Met Glu Arg Gly Pro Gln Thr 
        35                  40                  45              


Leu Lys Glu Thr Ser Phe Asn Gln Ala Tyr Gly Arg Asp Leu Met Glu 
    50                  55                  60                  


Ala Gln Glu Trp Cys Arg Lys Tyr Met Lys Ser Gly Asn Val Lys Asp 
65                  70                  75                  80  


Leu His Gln Ala Phe Asp Leu Tyr Tyr His Val Phe Arg Arg Ile Ser 
                85                  90                  95      


Lys Leu Glu 
            


<210>  9
<211>  100
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  chimeric protein heterodimerization domain, FRB segment of mTOR 
       with K to P substitution at residue 2095 of the full mTOR

<400>  9

Met Ala Ser Arg Ile Leu Trp His Glu Met Trp His Glu Gly Leu Glu 
1               5                   10                  15      


Glu Ala Ser Arg Leu Tyr Phe Gly Glu Arg Asn Val Lys Gly Met Phe 
            20                  25                  30          


Glu Val Leu Glu Pro Leu His Ala Met Met Glu Arg Gly Pro Gln Thr 
        35                  40                  45              


Leu Lys Glu Thr Ser Phe Asn Gln Ala Tyr Gly Arg Asp Leu Met Glu 
    50                  55                  60                  


Ala Gln Glu Trp Cys Arg Lys Tyr Met Lys Ser Gly Asn Val Pro Asp 
65                  70                  75                  80  


Leu Thr Gln Ala Trp Asp Leu Tyr Tyr His Val Phe Arg Arg Ile Ser 
                85                  90                  95      


Lys Leu Glu Ser 
            100 


<210>  10
<211>  20
<212>  PRT
<213>  Foot-and-mouth disease virus

<400>  10

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


Asn Pro Gly Pro 
            20  


<210>  11
<211>  20
<212>  PRT
<213>  Foot-and-mouth disease virus

<400>  11

Gln Cys Thr Asn Tyr Ala Leu Leu Lys Leu Ala Gly Asp Val Glu Ser 
1               5                   10                  15      


Asn Pro Gly Pro 
            20  


<210>  12
<211>  517
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  RapCasp9 suicide gene

<400>  12

Met Ala Ser Arg Ile Leu Trp His Glu Met Trp His Glu Gly Leu Glu 
1               5                   10                  15      


Glu Ala Ser Arg Leu Tyr Phe Gly Glu Arg Asn Val Lys Gly Met Phe 
            20                  25                  30          


Glu Val Leu Glu Pro Leu His Ala Met Met Glu Arg Gly Pro Gln Thr 
        35                  40                  45              


Leu Lys Glu Thr Ser Phe Asn Gln Ala Tyr Gly Arg Asp Leu Met Glu 
    50                  55                  60                  


Ala Gln Glu Trp Cys Arg Lys Tyr Met Lys Ser Gly Asn Val Lys Asp 
65                  70                  75                  80  


Leu Leu Gln Ala Trp Asp Leu Tyr Tyr His Val Phe Arg Arg Ile Ser 
                85                  90                  95      


Lys Leu Glu Tyr Ser Gly Gly Gly Ser Leu Glu Gly Val Gln Val Glu 
            100                 105                 110         


Thr Ile Ser Pro Gly Asp Gly Arg Thr Phe Pro Lys Arg Gly Gln Thr 
        115                 120                 125             


Cys Val Val His Tyr Thr Gly Met Leu Glu Asp Gly Lys Lys Phe Asp 
    130                 135                 140                 


Ser Ser Arg Asp Arg Asn Lys Pro Phe Lys Phe Met Leu Gly Lys Gln 
145                 150                 155                 160 


Glu Val Ile Arg Gly Trp Glu Glu Gly Val Ala Gln Met Ser Val Gly 
                165                 170                 175     


Gln Arg Ala Lys Leu Thr Ile Ser Pro Asp Tyr Ala Tyr Gly Ala Thr 
            180                 185                 190         


Gly His Pro Gly Ile Ile Pro Pro His Ala Thr Leu Val Phe Asp Val 
        195                 200                 205             


Glu Leu Leu Lys Leu Glu Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 
    210                 215                 220                 


Ser Gly Gly Gly Gly Ser Gly Val Asp Gly Phe Gly Asp Val Gly Ala 
225                 230                 235                 240 


Leu Glu Ser Leu Arg Gly Asn Ala Asp Leu Ala Tyr Ile Leu Ser Met 
                245                 250                 255     


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


Glu Ser Gly Leu Arg Thr Arg Thr Gly Ser Asn Ile Asp Cys Glu Lys 
        275                 280                 285             


Leu Arg Arg Arg Phe Ser Ser Leu His Phe Met Val Glu Val Lys Gly 
    290                 295                 300                 


Asp Leu Thr Ala Lys Lys Met Val Leu Ala Leu Leu Glu Leu Ala Gln 
305                 310                 315                 320 


Gln Asp His Gly Ala Leu Asp Cys Cys Val Val Val Ile Leu Ser His 
                325                 330                 335     


Gly Cys Gln Ala Ser His Leu Gln Phe Pro Gly Ala Val Tyr Gly Thr 
            340                 345                 350         


Asp Gly Cys Pro Val Ser Val Glu Lys Ile Val Asn Ile Phe Asn Gly 
        355                 360                 365             


Thr Ser Cys Pro Ser Leu Gly Gly Lys Pro Lys Leu Phe Phe Ile Gln 
    370                 375                 380                 


Ala Cys Gly Gly Glu Gln Lys Asp His Gly Phe Glu Val Ala Ser Thr 
385                 390                 395                 400 


Ser Pro Glu Asp Glu Ser Pro Gly Ser Asn Pro Glu Pro Asp Ala Thr 
                405                 410                 415     


Pro Phe Gln Glu Gly Leu Arg Thr Phe Asp Gln Leu Asp Ala Ile Ser 
            420                 425                 430         


Ser Leu Pro Thr Pro Ser Asp Ile Phe Val Ser Tyr Ser Thr Phe Pro 
        435                 440                 445             


Gly Phe Val Ser Trp Arg Asp Pro Lys Ser Gly Ser Trp Tyr Val Glu 
    450                 455                 460                 


Thr Leu Asp Asp Ile Phe Glu Gln Trp Ala His Ser Glu Asp Leu Gln 
465                 470                 475                 480 


Ser Leu Leu Leu Arg Val Ala Asn Ala Val Ser Val Lys Gly Ile Tyr 
                485                 490                 495     


Lys Gln Met Pro Gly Cys Phe Asn Phe Leu Arg Lys Lys Leu Phe Phe 
            500                 505                 510         


Lys Thr Ser Ala Ser 
        515         


<210>  13
<211>  5
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  linker

<400>  13

Ser Gly Gly Gly Ser 
1               5   


