                         LISTAGEM DE SEQUNCIAS

<110>  FUNDAO OSWALDO CRUZ
 
<120>  "POLIPEPTDEO COM ATIVIDADE ASPARAGINASE, POLINUCLEOTDEO, CASSETE DE EXPRESSO, 
       VETOR DE EXPRESSO, CLULA HOSPEDEIRA, COMPOSIO FARMACUTICA, 
       MTODOS PARA PRODUZIR UM POLIPEPTDEO COM ATIVIDADE ASPARAGINASE 
       E PARA PREVENIR OU TRATAR NEOPLASIAS, E, USO DE UM POLIPEPTDEO"

<130>  R000607

<160>  12    

<170>  PatentIn version 3.5

<210>  1
<211>  326
<212>  PRT
<213>  Escherichia coli

<400>  1

Leu Pro Asn Ile Thr Ile Leu Ala Thr Gly Gly Thr Ile Ala Gly Gly 
1               5                   10                  15      


Gly Asp Ser Ala Thr Lys Ser Asn Tyr Thr Ala Gly Lys Val Gly Val 
            20                  25                  30          


Glu Asn Leu Val Asn Ala Val Pro Gln Leu Lys Asp Ile Ala Asn Val 
        35                  40                  45              


Lys Gly Glu Gln Val Val Asn Ile Gly Ser Gln Asp Met Asn Asp Asn 
    50                  55                  60                  


Val Trp Leu Thr Leu Ala Lys Lys Ile Asn Thr Asp Cys Asp Lys Thr 
65                  70                  75                  80  


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


Tyr Phe Leu Asp Leu Thr Val Lys Cys Asp Lys Pro Val Val Met Val 
            100                 105                 110         


Gly Ala Met Arg Pro Ser Thr Ser Met Ser Ala Asp Gly Pro Phe Asn 
        115                 120                 125             


Leu Tyr Asn Ala Val Val Thr Ala Ala Asp Lys Ala Ser Ala Asn Arg 
    130                 135                 140                 


Gly Val Leu Val Val Met Asn Asp Thr Val Leu Asp Gly Arg Asp Val 
145                 150                 155                 160 


Thr Lys Thr Asn Thr Thr Asp Val Ala Thr Phe Lys Ser Val Asn Tyr 
                165                 170                 175     


Gly Pro Leu Gly Tyr Ile His Asn Gly Lys Ile Asp Tyr Gln Arg Thr 
            180                 185                 190         


Pro Ala Arg Lys His Thr Ser Asp Thr Pro Phe Asp Val Ser Lys Leu 
        195                 200                 205             


Asn Glu Leu Pro Lys Val Gly Ile Val Tyr Asn Tyr Ala Asn Ala Ser 
    210                 215                 220                 


Asp Leu Pro Ala Lys Ala Leu Val Asp Ala Gly Tyr Asp Gly Ile Val 
225                 230                 235                 240 


Ser Ala Gly Val Gly Asn Gly Asn Leu Tyr Lys Ser Val Phe Asp Thr 
                245                 250                 255     


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


Val Pro Thr Gly Ala Thr Thr Gln Asp Ala Glu Val Asp Asp Ala Lys 
        275                 280                 285             


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


Leu Leu Gln Leu Ala Leu Thr Gln Thr Lys Asp Pro Gln Gln Ile Gln 
305                 310                 315                 320 


Gln Ile Phe Asn Gln Tyr 
                325     


<210>  2
<211>  324
<212>  PRT
<213>  Erwinia carotovora

<400>  2

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


Ala Ala Ala Asn Thr Gln Thr Thr Gly Tyr Lys Ala Gly Ala Leu Gly 
            20                  25                  30          


Val Glu Thr Leu Ile Gln Ala Val Pro Glu Leu Lys Thr Leu Ala Asn 
        35                  40                  45              


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


Asp Val Leu Leu Thr Leu Ser Lys Arg Val Asn Glu Leu Leu Ala Arg 
65                  70                  75                  80  


Ser Asp Val Asp Gly Val Val Ile Thr His Gly Thr Asp Thr Leu Asp 
                85                  90                  95      


Glu Ser Pro Tyr Phe Leu Asn Leu Thr Val Lys Ser Asp Lys Pro Val 
            100                 105                 110         


Val Phe Val Ala Ala Met Arg Pro Ala Thr Ala Ile Ser Ala Asp Gly 
        115                 120                 125             


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


Arg Gly Arg Gly Val Leu Val Val Leu Asn Asp Arg Ile Gly Ser Ala 
145                 150                 155                 160 


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


Pro Glu Glu Gly Tyr Leu Gly Val Ile Ile Gly Asp Lys Ile Tyr Tyr 
            180                 185                 190         


Gln Thr Arg Leu Asp Lys Val His Thr Thr Arg Ser Val Phe Asp Val 
        195                 200                 205             


Thr Asn Val Asp Lys Leu Pro Ala Val Asp Ile Ile Tyr Gly Tyr Gln 
    210                 215                 220                 


Asp Asp Pro Glu Tyr Met Tyr Asp Ala Ser Ile Lys His Gly Val Lys 
225                 230                 235                 240 


Gly Ile Val Tyr Ala Gly Met Gly Ala Gly Ser Val Ser Lys Arg Gly 
                245                 250                 255     


Asp Ala Gly Ile Arg Lys Ala Glu Ser Lys Gly Ile Val Val Val Arg 
            260                 265                 270         


Ser Ser Arg Thr Gly Ser Gly Ile Val Pro Pro Asp Ala Gly Gln Pro 
        275                 280                 285             


Gly Leu Val Ala Asp Ser Leu Ser Pro Ala Lys Ser Arg Ile Leu Leu 
    290                 295                 300                 


Met Leu Ala Leu Thr Lys Thr Thr Asn Pro Ala Val Ile Gln Asp Tyr 
305                 310                 315                 320 


Phe His Ala Tyr 
                


<210>  3
<211>  326
<212>  PRT
<213>  Sequncia Artificial

<220>
<223>  Sequncia de aminocidos da asparaginase mutada de E. coli

<400>  3

Leu Pro Asn Ile Thr Ile Leu Ala Thr Gly Gly Thr Ile Ala Gly Gly 
1               5                   10                  15      


Gly Asp Ser Ala Thr Lys Ser Asn Tyr Thr Ala Gly Lys Val Gly Val 
            20                  25                  30          


Glu Asn Leu Val Asn Ala Val Pro Gln Leu Lys Asp Ile Ala Asn Val 
        35                  40                  45              


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


Val Leu Leu Lys Leu Ser Lys Arg Val Asn Glu Asp Cys Asp Lys Thr 
65                  70                  75                  80  


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


Tyr Phe Leu Asp Leu Thr Val Lys Cys Asp Lys Pro Val Val Met Val 
            100                 105                 110         


Gly Ala Met Arg Pro Ser Thr Ser Met Ser Ala Asp Gly Pro Phe Asn 
        115                 120                 125             


Leu Tyr Asn Ala Val Val Thr Ala Ala Asp Lys Ala Ser Ala Asn Arg 
    130                 135                 140                 


Gly Val Leu Val Val Met Asn Asp Thr Val Leu Asp Gly Arg Asp Val 
145                 150                 155                 160 


Thr Lys Thr Asn Thr Thr Asp Val Ala Thr Phe Lys Ser Val Asn Tyr 
                165                 170                 175     


Gly Pro Leu Gly Tyr Ile His Asn Gly Lys Ile Asp Tyr Gln Arg Thr 
            180                 185                 190         


Pro Ala Arg Lys His Thr Ser Asp Thr Pro Phe Asp Val Ser Lys Leu 
        195                 200                 205             


Asn Glu Leu Pro Lys Val Gly Ile Val Tyr Asn Tyr Ala Asn Ala Ser 
    210                 215                 220                 


Asp Leu Pro Ala Lys Ala Leu Val Asp Ala Gly Tyr Asp Gly Ile Val 
225                 230                 235                 240 


Ser Ala Gly Val Gly Asn Gly Asn Leu Tyr Lys Ser Val Phe Asp Thr 
                245                 250                 255     


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


Val Pro Thr Gly Ala Thr Thr Gln Asp Ala Glu Val Asp Asp Ala Lys 
        275                 280                 285             


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


Leu Leu Gln Leu Ala Leu Thr Gln Thr Lys Asp Pro Gln Gln Ile Gln 
305                 310                 315                 320 


Gln Ile Phe Asn Gln Tyr 
                325     


<210>  4
<211>  984
<212>  DNA
<213>  Sequncia Artificial

<220>
<223>  Sequncia de nucleotdeos da asparagina mutada de E.coli

<400>  4
catatgttac ccaatatcac cattttagca accggcggga ccattgccgg tggtggtgac       60

tccgcaacca aatctaacta cacagcgggt aaagttggcg tagaaaatct ggttaatgcg      120

gtgccgcaac tgaaggacat tgcgaacgtt aaaggcgagc aggtcgcttc gatcggctcc      180

gaaaatatga cctctgatgt tctgctgaaa ctgagtaaac gtgtcaacga agattgcgac      240

aaaaccgatg gttttgtgat tacccatggc accgacacga tggaagaaac cgcttacttc      300

ctcgacctga cggtgaaatg cgacaaaccg gtggtgatgg tcggtgcaat gcgtccgtcc      360

acgtctatga gcgcagacgg tccattcaac ctgtataacg cggtagtgac tgcagctgat      420

aaagcctccg ctaatcgtgg cgtactggta gtgatgaacg acaccgtgct tgatggccgt      480

gatgtcacca aaaccaacac caccgatgta gcgaccttca agtctgttaa ctacggtcct      540

ctgggttaca ttcacaacgg taagattgac taccaacgta ccccggcacg taagcacacc      600

agcgacacgc cgttcgatgt ctctaagctg aatgaactgc cgaaagtcgg cattgtttat      660

aactacgcta acgcatccga tcttccggct aaagcactgg tagatgcggg ctatgatggc      720

atcgttagcg ctggcgtggg taacggcaac ctgtataaat ccgtatttga cacccttgca      780

accgctgcga aaaacggcac tgcagtagtg cgttcttccc gcgtaccgac gggcgctacc      840

actcaggatg ccgaagtgga tgatgcgaaa tacggcttcg tcgcctctgg cacgttgaac      900

ccgcaaaaag cgcgcgttct gctgcaactg gctctgacgc aaactaaaga tccgcagcag      960

atccaacaaa tcttcaatca gtac                                             984


<210>  5
<211>  980
<212>  DNA
<213>  Escherichia coli

<400>  5
gcttacccaa tatcaccatt ttagcaaccg gcgggaccat tgccggtggt ggtgactccg       60

caaccaaatc taactacaca gcgggtaaag ttggcgtaga aaatctggtt aatgcggtgc      120

cgcaactaaa agacattgcg aacgttaaag gcgagcaggt agtgaatatc ggctcccagg      180

acatgaacga taatgtctgg ctgacactgg cgaaaaaaat taacaccgac tgcgataaaa      240

ccgacggctt cgtcattacc cacggtaccg acacgatgga agaaaccgct tacttcctcg      300

acctgacggt gaaatgcgac aaaccggtgg tgatggtcgg cgcaatgcgc ccgtccacgt      360

ccatgagcgc agacggtcca ttcaacctgt ataacgcggt agtgaccgca gctgataaag      420

cctccgctaa tcgtggcgtg ctggtggtga tgaacgacac cgtactggac ggtcgcgatg      480

tcaccaaaac caacaccacc gacgtagcga ccttcaagtc tgttaactac ggtcctctgg      540

gatacattca caacggtaag attgactacc aacgtacccc ggcacgtaag cacaccagcg      600

atacgccatt cgatgtctct aagctgaatg agctgccgaa agtcggcatc gtttataact      660

acgctaacgc atccgatctt ccggctaaag cactggtaga tgcgggctat gatggcatcg      720

ttagcgctgg tgtgggtaat ggtaacctgt ataaatccgt gttcgacacc ctggcaaccg      780

ccgcgaaaaa cggcactgca gtagtgcgtt cttcccgcgt accgacgggt gctaccactc      840

aggatgctga agtggatgat gcgaaatacg gcttcgtcgc ctctggcacg ctgaacccgc      900

aaaaagcgcg cgtcctgctg cagctggctc tgacgcaaac caaagatccg cagcagatcc      960

agcagatctt caatcagtac                                                  980


<210>  6
<211>  975
<212>  DNA
<213>  Erwinia carotovora

<400>  6
aacctaccta atattgttat tctggcaacg ggcggtacga ttgccggttc cgctgcggcc       60

aatacgcaaa ccacggggta taaagcgggt gcgctgggcg tagagacgct gattcaagcg      120

gtgccagagc tgaaaacgct tgccaatatt tcaggcgagc aggttgccag catcggcagt      180

gaaaatatga ccagcgatgt tctgctgaag ctgagcaagc gcgtgaacga gctgctggca      240

cgcagcgatg ttgatggcgt ggtgattacg cacggtacgg atacgctcga cgaatcgcct      300

tacttcctga acctgacggt gaagagtgac aaaccggtgg tctttgccgc cgcgatgcgt      360

ccggcaacgg caatcagtgc tgacggcccg atgaacctgt acggtgcagt aaaagtggca      420

gcggataaaa actctcgcgg ccgcggcgtg ctggtcgtgc ttaacgaccg tattggttct      480

gctcgtttca tcagcaaaac caacgcttct acgctggata cctttaaagc gccagaagaa      540

ggctatctgg gcgtgattat cggtgacaaa gtttactacc agacgcgtct ggataaagtt      600

cacaccacgc gttccgtatt tgacgtgacc aacgtcgata aactgccagc cgttgacatt      660

atttatggct atcaggacga tccagaatac atgtatgacg ccgccatcaa acacggcgcg      720

aaagggattg tgtatgcggg catgggcgca ggtagcgtat ccaaacgcgg cgatgctggc      780

atccgtaaag cggaaagcaa aggcgttgtt gttgtacgct ccagccgtac cggcagcggc      840

atcgtaccac cggatgcggg ccagcctggt ctggttgctg attctctgag cccagctaaa      900

tcacgtattc tgctgatgct ggcgctgaca aaaacgacga accccgcggt gattcaggat      960

tatttccacg cgtat                                                       975


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

<220>
<223>  Iniciador frag 1/2 direto

<400>  7
catatgttac ccaatatcac cattttagca                                        30


<210>  8
<211>  26
<212>  DNA
<213>  Sequncia Artificial

<220>
<223>  Iniciador frag 1/2 reverso

<400>  8
catgccatgg gtaatcacaa accatc                                            26


<210>  9
<211>  28
<212>  DNA
<213>  Sequncia Artificial

<220>
<223>  Iniciador frag 3 direto

<400>  9
catgccatgg caccgacacg atggaaga                                          28


<210>  10
<211>  27
<212>  DNA
<213>  Sequncia Artificial

<220>
<223>  Iniciador frag 3 reverso

<400>  10
ctcgagtcag tattgattga agatttg                                           27


<210>  11
<211>  27
<212>  DNA
<213>  Sequncia Artificial

<220>
<223>  Iniciador Ecar-LANS direto

<400>  11
catatgaacc tgccgaacat tgtgatt                                           27


<210>  12
<211>  30
<212>  DNA
<213>  Sequncia Artificial

<220>
<223>  Iniciador Ecar-LANS reverso

<400>  12
ctggagttag taagcgtgga agtagtcttg                                        30


