                         SEQUENCE LISTING

<110>  Children's Hospital of Los Angeles
 
<120>  NOVEL ASPARAGINASE AND METHODS FOR TREATING DISEASES ASSOCIATED 
       WITH ASPARAGINE DEPENDENCE

<130>  043833.019

<150>  61/545,238
<151>  2011-10-10

<160>  10    

<170>  PatentIn version 3.5

<210>  1
<211>  331
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Derived from W. succinogenes

<400>  1

Met Met Ala Lys Pro Gln Val Thr Ile Leu Ala Thr Gly Gly Thr Ile 
1               5                   10                  15      


Ala Gly Ser Gly Glu Ser Ser Val Lys Ser Ser Tyr Ser Ala Gly Ala 
            20                  25                  30          


Val Thr Val Asp Lys Leu Leu Ala Ala Val Pro Ala Ile Asn Asp Leu 
        35                  40                  45              


Ala Thr Ile Lys Gly Glu Gln Ile Ser Ser Ile Gly Ser Gln Glu Met 
    50                  55                  60                  


Thr Gly Lys Val Trp Leu Lys Leu Ala Lys Arg Val Asn Glu Leu Leu 
65                  70                  75                  80  


Ala Gln Lys Glu Thr Glu Ala Val Ile Ile Thr His Gly Thr Asp Thr 
                85                  90                  95      


Met Glu Glu Thr Ala Phe Phe Leu Asn Leu Thr Val Lys Ser Gln Lys 
            100                 105                 110         


Pro Val Val Leu Val Gly Ala Met Arg Ser Gly Ser Ser Met Ser Ala 
        115                 120                 125             


Asp Gly Pro Met Asn Leu Tyr Asn Ala Val Asn Val Ala Ile Asn Lys 
    130                 135                 140                 


Ala Ser Thr Asn Lys Gly Val Val Ile Val Met Asn Asp Glu Ile His 
145                 150                 155                 160 


Ala Ala Arg Glu Ala Thr Lys Leu Asn Thr Thr Ala Val Asn Ala Phe 
                165                 170                 175     


Ala Ser Pro Asn Thr Gly Lys Ile Gly Thr Val Tyr Tyr Gly Lys Val 
            180                 185                 190         


Glu Tyr Phe Thr Gln Ser Val Arg Pro His Thr Leu Ala Ser Glu Phe 
        195                 200                 205             


Asp Ile Ser Lys Ile Glu Glu Leu Pro Arg Val Asp Ile Leu Tyr Ala 
    210                 215                 220                 


His Pro Asp Asp Thr Asp Val Leu Val Asn Ala Ala Leu Gln Ala Gly 
225                 230                 235                 240 


Ala Lys Gly Ile Ile His Ala Gly Met Gly Asn Gly Asn Pro Phe Pro 
                245                 250                 255     


Leu Thr Gln Asn Ala Leu Glu Lys Ala Ala Lys Ser Gly Val Val Val 
            260                 265                 270         


Ala Arg Ser Ser Arg Val Gly Ser Gly Ser Thr Thr Gln Glu Ala Glu 
        275                 280                 285             


Val Asp Asp Lys Lys Leu Gly Phe Val Ala Thr Glu Ser Leu Asn Pro 
    290                 295                 300                 


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


Arg Glu Ala Ile Gln Lys Ile Phe Ser Thr Tyr 
                325                 330     


<210>  2
<211>  330
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Derived from W. succinogenes

<400>  2

Met Ala Lys Pro Gln Val Thr Ile Leu Ala Thr Gly Gly Thr Ile Ala 
1               5                   10                  15      


Gly Ser Gly Glu Ser Ser Val Lys Ser Ser Tyr Ser Ala Gly Ala Val 
            20                  25                  30          


Thr Val Asp Lys Leu Leu Ala Ala Val Pro Ala Ile Asn Asp Leu Ala 
        35                  40                  45              


Thr Ile Lys Gly Glu Gln Ile Ser Ser Ile Gly Ser Gln Glu Met Thr 
    50                  55                  60                  


Gly Lys Val Trp Leu Lys Leu Ala Lys Arg Val Asn Glu Leu Leu Ala 
65                  70                  75                  80  


Gln Lys Glu Thr Glu Ala Val Ile Ile Thr His Gly Thr Asp Thr Met 
                85                  90                  95      


Glu Glu Thr Ala Phe Phe Leu Asn Leu Thr Val Lys Ser Gln Lys Pro 
            100                 105                 110         


Val Val Leu Val Gly Ala Met Arg Ser Gly Ser Ser Met Ser Ala Asp 
        115                 120                 125             


Gly Pro Met Asn Leu Tyr Asn Ala Val Asn Val Ala Ile Asn Lys Ala 
    130                 135                 140                 


Ser Thr Asn Lys Gly Val Val Ile Val Met Asn Asp Glu Ile His Ala 
145                 150                 155                 160 


Ala Arg Glu Ala Thr Lys Leu Asn Thr Thr Ala Val Asn Ala Phe Ala 
                165                 170                 175     


Ser Pro Asn Thr Gly Lys Ile Gly Thr Val Tyr Tyr Gly Lys Val Glu 
            180                 185                 190         


Tyr Phe Thr Gln Ser Val Arg Pro His Thr Leu Ala Ser Glu Phe Asp 
        195                 200                 205             


Ile Ser Lys Ile Glu Glu Leu Pro Arg Val Asp Ile Leu Tyr Ala His 
    210                 215                 220                 


Pro Asp Asp Thr Asp Val Leu Val Asn Ala Ala Leu Gln Ala Gly Ala 
225                 230                 235                 240 


Lys Gly Ile Ile His Ala Gly Met Gly Asn Gly Asn Pro Phe Pro Leu 
                245                 250                 255     


Thr Gln Asn Ala Leu Glu Lys Ala Ala Lys Ser Gly Val Val Val Ala 
            260                 265                 270         


Arg Ser Ser Arg Val Gly Ser Gly Ser Thr Thr Gln Glu Ala Glu Val 
        275                 280                 285             


Asp Asp Lys Lys Leu Gly Phe Val Ala Thr Glu Ser Leu Asn Pro Gln 
    290                 295                 300                 


Lys Ala Arg Val Leu Leu Met Leu Ala Leu Thr Lys Thr Ser Asp Arg 
305                 310                 315                 320 


Glu Ala Ile Gln Lys Ile Phe Ser Thr Tyr 
                325                 330 


<210>  3
<211>  329
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Derived from W. succinogenes

<400>  3

Ala Lys Pro Gln Val Thr Ile Leu Ala Thr Gly Gly Thr Ile Ala Gly 
1               5                   10                  15      


Ser Gly Glu Ser Ser Val Lys Ser Ser Tyr Ser Ala Gly Ala Val Thr 
            20                  25                  30          


Val Asp Lys Leu Leu Ala Ala Val Pro Ala Ile Asn Asp Leu Ala Thr 
        35                  40                  45              


Ile Lys Gly Glu Gln Ile Ser Ser Ile Gly Ser Gln Glu Met Thr Gly 
    50                  55                  60                  


Lys Val Trp Leu Lys Leu Ala Lys Arg Val Asn Glu Leu Leu Ala Gln 
65                  70                  75                  80  


Lys Glu Thr Glu Ala Val Ile Ile Thr His Gly Thr Asp Thr Met Glu 
                85                  90                  95      


Glu Thr Ala Phe Phe Leu Asn Leu Thr Val Lys Ser Gln Lys Pro Val 
            100                 105                 110         


Val Leu Val Gly Ala Met Arg Ser Gly Ser Ser Met Ser Ala Asp Gly 
        115                 120                 125             


Pro Met Asn Leu Tyr Asn Ala Val Asn Val Ala Ile Asn Lys Ala Ser 
    130                 135                 140                 


Thr Asn Lys Gly Val Val Ile Val Met Asn Asp Glu Ile His Ala Ala 
145                 150                 155                 160 


Arg Glu Ala Thr Lys Leu Asn Thr Thr Ala Val Asn Ala Phe Ala Ser 
                165                 170                 175     


Pro Asn Thr Gly Lys Ile Gly Thr Val Tyr Tyr Gly Lys Val Glu Tyr 
            180                 185                 190         


Phe Thr Gln Ser Val Arg Pro His Thr Leu Ala Ser Glu Phe Asp Ile 
        195                 200                 205             


Ser Lys Ile Glu Glu Leu Pro Arg Val Asp Ile Leu Tyr Ala His Pro 
    210                 215                 220                 


Asp Asp Thr Asp Val Leu Val Asn Ala Ala Leu Gln Ala Gly Ala Lys 
225                 230                 235                 240 


Gly Ile Ile His Ala Gly Met Gly Asn Gly Asn Pro Phe Pro Leu Thr 
                245                 250                 255     


Gln Asn Ala Leu Glu Lys Ala Ala Lys Ser Gly Val Val Val Ala Arg 
            260                 265                 270         


Ser Ser Arg Val Gly Ser Gly Ser Thr Thr Gln Glu Ala Glu Val Asp 
        275                 280                 285             


Asp Lys Lys Leu Gly Phe Val Ala Thr Glu Ser Leu Asn Pro Gln Lys 
    290                 295                 300                 


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


Ala Ile Gln Lys Ile Phe Ser Thr Tyr 
                325                 


<210>  4
<211>  328
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Derived from W. succinogenes

<400>  4

Lys Pro Gln Val Thr Ile Leu Ala Thr Gly Gly Thr Ile Ala Gly Ser 
1               5                   10                  15      


Gly Glu Ser Ser Val Lys Ser Ser Tyr Ser Ala Gly Ala Val Thr Val 
            20                  25                  30          


Asp Lys Leu Leu Ala Ala Val Pro Ala Ile Asn Asp Leu Ala Thr Ile 
        35                  40                  45              


Lys Gly Glu Gln Ile Ser Ser Ile Gly Ser Gln Glu Met Thr Gly Lys 
    50                  55                  60                  


Val Trp Leu Lys Leu Ala Lys Arg Val Asn Glu Leu Leu Ala Gln Lys 
65                  70                  75                  80  


Glu Thr Glu Ala Val Ile Ile Thr His Gly Thr Asp Thr Met Glu Glu 
                85                  90                  95      


Thr Ala Phe Phe Leu Asn Leu Thr Val Lys Ser Gln Lys Pro Val Val 
            100                 105                 110         


Leu Val Gly Ala Met Arg Ser Gly Ser Ser Met Ser Ala Asp Gly Pro 
        115                 120                 125             


Met Asn Leu Tyr Asn Ala Val Asn Val Ala Ile Asn Lys Ala Ser Thr 
    130                 135                 140                 


Asn Lys Gly Val Val Ile Val Met Asn Asp Glu Ile His Ala Ala Arg 
145                 150                 155                 160 


Glu Ala Thr Lys Leu Asn Thr Thr Ala Val Asn Ala Phe Ala Ser Pro 
                165                 170                 175     


Asn Thr Gly Lys Ile Gly Thr Val Tyr Tyr Gly Lys Val Glu Tyr Phe 
            180                 185                 190         


Thr Gln Ser Val Arg Pro His Thr Leu Ala Ser Glu Phe Asp Ile Ser 
        195                 200                 205             


Lys Ile Glu Glu Leu Pro Arg Val Asp Ile Leu Tyr Ala His Pro Asp 
    210                 215                 220                 


Asp Thr Asp Val Leu Val Asn Ala Ala Leu Gln Ala Gly Ala Lys Gly 
225                 230                 235                 240 


Ile Ile His Ala Gly Met Gly Asn Gly Asn Pro Phe Pro Leu Thr Gln 
                245                 250                 255     


Asn Ala Leu Glu Lys Ala Ala Lys Ser Gly Val Val Val Ala Arg Ser 
            260                 265                 270         


Ser Arg Val Gly Ser Gly Ser Thr Thr Gln Glu Ala Glu Val Asp Asp 
        275                 280                 285             


Lys Lys Leu Gly Phe Val Ala Thr Glu Ser Leu Asn Pro Gln Lys Ala 
    290                 295                 300                 


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


Ile Gln Lys Ile Phe Ser Thr Tyr 
                325             


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

<220>
<223>  SRP signal sequence

<400>  5

Lys Lys Ile Trp Leu Ala Leu Ala Gly Leu Val Leu Ala Phe Ser Ala 
1               5                   10                  15      


Ser Ala 
        


<210>  6
<211>  25
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  MalE signal sequence

<400>  6

Lys Ile Lys Thr Gly Ala Arg Ile Leu Ala Leu Ser Ala Leu Thr Thr 
1               5                   10                  15      


Met Met Phe Ser Ala Ser Ala Leu Ala 
            20                  25  


<210>  7
<211>  30
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Forward W. succinogenes asparaginase-specific PCR primer

<400>  7
tccggatcca gcgcctctgt tttgatggct                                        30


<210>  8
<211>  30
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Reverse W. succinogenes asparaginase-specific PCR primer

<400>  8
tgggaattcg gtggagaaga tcttttggat                                        30


<210>  9
<211>  1133
<212>  DNA
<213>  Wolinella succinogenes


<220>
<221>  CDS
<222>  (118)..(1110)

<400>  9
atgggcagca gccatcatca tcatcatcat agcagcggcc tggtgccgcg cggcagccat       60

atggctagca tgactggtgg acagcaaatg ggtcgcggat ccagcgcctc tgttttg         117

atg gct aaa ccc caa gtg act atc cta gcc aca gga ggc acc atc gct        165
Met Ala Lys Pro Gln Val Thr Ile Leu Ala Thr Gly Gly Thr Ile Ala           
1               5                   10                  15                

ggt tcg ggg gaa tct agc gtc aag agt agc tac tct gct gga gca gtc        213
Gly Ser Gly Glu Ser Ser Val Lys Ser Ser Tyr Ser Ala Gly Ala Val           
            20                  25                  30                    

acc gtt gat aag ctt ctt gca gcc gtc cct gcc atc aac gac cta gcc        261
Thr Val Asp Lys Leu Leu Ala Ala Val Pro Ala Ile Asn Asp Leu Ala           
        35                  40                  45                        

acc atc aag ggt gaa cag atc tca agc att ggc tcc caa gag atg acg        309
Thr Ile Lys Gly Glu Gln Ile Ser Ser Ile Gly Ser Gln Glu Met Thr           
    50                  55                  60                            

ggt aag gtg tgg ctt aaa cta gcc aag cgt gtc aat gag ctc ctc gcc        357
Gly Lys Val Trp Leu Lys Leu Ala Lys Arg Val Asn Glu Leu Leu Ala           
65                  70                  75                  80            

caa aaa gag acc gaa gcc gtg atc atc acc cat gga act gac acc atg        405
Gln Lys Glu Thr Glu Ala Val Ile Ile Thr His Gly Thr Asp Thr Met           
                85                  90                  95                

gaa gag acc gct ttc ttc ctc aac ctc acg gtg aaa agc caa aaa cct        453
Glu Glu Thr Ala Phe Phe Leu Asn Leu Thr Val Lys Ser Gln Lys Pro           
            100                 105                 110                   

gtc gtc ctt gta ggc gcc atg cgt cca ggc tct tcc atg agt gct gat        501
Val Val Leu Val Gly Ala Met Arg Pro Gly Ser Ser Met Ser Ala Asp           
        115                 120                 125                       

ggc ccc atg aat ctc tat aac gcc gtg aat gta gcg atc aac aaa gcc        549
Gly Pro Met Asn Leu Tyr Asn Ala Val Asn Val Ala Ile Asn Lys Ala           
    130                 135                 140                           

tct act aac aaa gga gtg gtg att gtg atg aac gat gag att cac gcc        597
Ser Thr Asn Lys Gly Val Val Ile Val Met Asn Asp Glu Ile His Ala           
145                 150                 155                 160           

gcc aga gaa gcg acc aag ctc aac acc acc gca gtc aat gca ttt gct        645
Ala Arg Glu Ala Thr Lys Leu Asn Thr Thr Ala Val Asn Ala Phe Ala           
                165                 170                 175               

tcg ccc aac aca ggt aaa atc ggc aca gtc tat tat ggc aaa gtc gag        693
Ser Pro Asn Thr Gly Lys Ile Gly Thr Val Tyr Tyr Gly Lys Val Glu           
            180                 185                 190                   

tat ttc act caa tcc gtt cga cct cac acc ctt gca agt gag ttt gat        741
Tyr Phe Thr Gln Ser Val Arg Pro His Thr Leu Ala Ser Glu Phe Asp           
        195                 200                 205                       

att agc aaa atc gaa gaa ctc ccc aga gtc gat att ctt tac gct cac        789
Ile Ser Lys Ile Glu Glu Leu Pro Arg Val Asp Ile Leu Tyr Ala His           
    210                 215                 220                           

ccc gat gat act gat gtt tta gtc aat gca gcc ctt cag gca gga gcc        837
Pro Asp Asp Thr Asp Val Leu Val Asn Ala Ala Leu Gln Ala Gly Ala           
225                 230                 235                 240           

aaa gga atc atc cat gca ggc atg ggc aat ggg aac cct ttc cct ttg        885
Lys Gly Ile Ile His Ala Gly Met Gly Asn Gly Asn Pro Phe Pro Leu           
                245                 250                 255               

act caa aat gct ctt gaa aaa gca gcc aaa tca ggc gta gtc gtc gct        933
Thr Gln Asn Ala Leu Glu Lys Ala Ala Lys Ser Gly Val Val Val Ala           
            260                 265                 270                   

cga agc tct aga gtg ggc agt ggt tcc acc acc caa gag gct gaa gtg        981
Arg Ser Ser Arg Val Gly Ser Gly Ser Thr Thr Gln Glu Ala Glu Val           
        275                 280                 285                       

gat gat aag aaa ctt ggt ttt gtg gct aca gag agt ctc aac cct caa       1029
Asp Asp Lys Lys Leu Gly Phe Val Ala Thr Glu Ser Leu Asn Pro Gln           
    290                 295                 300                           

aaa gcc aga gtg ctt ctt atg tta gcc ctc acc aaa act agt gat aga       1077
Lys Ala Arg Val Leu Leu Met Leu Ala Leu Thr Lys Thr Ser Asp Arg           
305                 310                 315                 320           

gag gcg atc caa aag atc ttc tcc acc tat taa tccaagaaag ggaatctctt     1130
Glu Ala Ile Gln Lys Ile Phe Ser Thr Tyr                                   
                325                 330                                   

cac                                                                   1133


<210>  10
<211>  330
<212>  PRT
<213>  Wolinella succinogenes

<400>  10

Met Ala Lys Pro Gln Val Thr Ile Leu Ala Thr Gly Gly Thr Ile Ala 
1               5                   10                  15      


Gly Ser Gly Glu Ser Ser Val Lys Ser Ser Tyr Ser Ala Gly Ala Val 
            20                  25                  30          


Thr Val Asp Lys Leu Leu Ala Ala Val Pro Ala Ile Asn Asp Leu Ala 
        35                  40                  45              


Thr Ile Lys Gly Glu Gln Ile Ser Ser Ile Gly Ser Gln Glu Met Thr 
    50                  55                  60                  


Gly Lys Val Trp Leu Lys Leu Ala Lys Arg Val Asn Glu Leu Leu Ala 
65                  70                  75                  80  


Gln Lys Glu Thr Glu Ala Val Ile Ile Thr His Gly Thr Asp Thr Met 
                85                  90                  95      


Glu Glu Thr Ala Phe Phe Leu Asn Leu Thr Val Lys Ser Gln Lys Pro 
            100                 105                 110         


Val Val Leu Val Gly Ala Met Arg Pro Gly Ser Ser Met Ser Ala Asp 
        115                 120                 125             


Gly Pro Met Asn Leu Tyr Asn Ala Val Asn Val Ala Ile Asn Lys Ala 
    130                 135                 140                 


Ser Thr Asn Lys Gly Val Val Ile Val Met Asn Asp Glu Ile His Ala 
145                 150                 155                 160 


Ala Arg Glu Ala Thr Lys Leu Asn Thr Thr Ala Val Asn Ala Phe Ala 
                165                 170                 175     


Ser Pro Asn Thr Gly Lys Ile Gly Thr Val Tyr Tyr Gly Lys Val Glu 
            180                 185                 190         


Tyr Phe Thr Gln Ser Val Arg Pro His Thr Leu Ala Ser Glu Phe Asp 
        195                 200                 205             


Ile Ser Lys Ile Glu Glu Leu Pro Arg Val Asp Ile Leu Tyr Ala His 
    210                 215                 220                 


Pro Asp Asp Thr Asp Val Leu Val Asn Ala Ala Leu Gln Ala Gly Ala 
225                 230                 235                 240 


Lys Gly Ile Ile His Ala Gly Met Gly Asn Gly Asn Pro Phe Pro Leu 
                245                 250                 255     


Thr Gln Asn Ala Leu Glu Lys Ala Ala Lys Ser Gly Val Val Val Ala 
            260                 265                 270         


Arg Ser Ser Arg Val Gly Ser Gly Ser Thr Thr Gln Glu Ala Glu Val 
        275                 280                 285             


Asp Asp Lys Lys Leu Gly Phe Val Ala Thr Glu Ser Leu Asn Pro Gln 
    290                 295                 300                 


Lys Ala Arg Val Leu Leu Met Leu Ala Leu Thr Lys Thr Ser Asp Arg 
305                 310                 315                 320 


Glu Ala Ile Gln Lys Ile Phe Ser Thr Tyr 
                325                 330 


