                         SEQUENCE LISTING

<110>  INTERNATIONAL CENTRE FOR GENETIC ENGINEERING AND 
       BIOTECHNOLOGY (ICGEB)
 
<120>  RECOMBINANT PROTEINS WITH A SELECTIVE INACTIVATION ACTIVITY ON 
       TARGET PROTEINS

<130>  RBW09126

<150>  PD2010A000356
<151>  2010-11-25

<160>  10    

<170>  PatentIn version 3.3

<210>  1
<211>  12
<212>  PRT
<213>  Artificial

<220>
<223>  SV5 tag

<400>  1

Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu Asp 
1               5                   10          


<210>  2
<211>  2035
<212>  DNA
<213>  Artificial

<220>
<223>  scFv9E1-degradin-gene

<400>  2
atgggctgga gcctgatcct cctgttcctc gtcgctgtgg ctacaggtaa ggggctcaca     60

gtagcaggct tgaggtctgg acatatatat gggtgacaat gacatccact ttgcctttct    120

ctccacaggt gtgcactctc aaattgtgct gacccagtct ccagcaatca tgtctgcatc    180

tccaggggag agggtcacca taacctgcag tgtcagttca actgtaagct acatgcactg    240

gttccagcag aagccaggca cttctcccaa agtcttgatt catgacacat ccaagttggc    300

ttctggagtc cctgctcgct tcagtggcag tggttctggg acctcttact ctctcacaat    360

cagccgaatg gaggctgaag atgctgccac ttattactgc cagcaaagga gttatttccc    420

gctcacgttc ggtattggga ccaagctgga gctgaaaggc agcactagtg gtagcggcaa    480

accaggttcc ggcgaaggct cgagcaaagg ccaggtgcag ctgaaggagt cgggacctgg    540

cctggtggcg ccctcacaga gtctgtccat cacttgcact gtctctggat tttcattaac    600

cagttatggt atacactgga ttcgccagcc tccaggaaag ggtctggagt ggctgggagt    660

aatatgggct ggtggaaaca caagttataa ttcggctctc atgtccagac tgagcatcag    720

caaagacaac tccaggagcc aagttatctt acaaatgaac agtctgcaaa ctgatgacac    780

agccatatac tactgtgcca gagaggaccg gatgtattgg tacttcgatg tctggggcgc    840

agggaccacg gtcaccgtct cctccggagc tagtggcaaa ccaatcccaa acccactgct    900

gggcctggat gctagcggaa gtgacattgt acctcagagt aatgagacag ctctccacta    960

ctttaagaaa gctgctgaca tgggcaaccc agttggacag agtgggcttg gaatggccta   1020

cctctatggg agaggagttc aagttaatta tgatctagcc cttaagtatt tccagaaagc   1080

tgctgaacaa ggctgggtgg atgggcagct acagcttggt tccatgtact ataatggcat   1140

tggagtcaag agagattata aacaggcctt gaagtatttt aatttagctt ctcagggagg   1200

ccatatcttg gctttctata acctagctca gatgcatgcc agtggcaccg gcgtgatgcg   1260

atcatgtcac actgcagtgg agttgtttaa gaatgtatgt gaacgaggcc gttggtctga   1320

aaggcttatg actgcctata acagctataa agatggcgat tacaatgctg cagtgatcca   1380

gtacctcctc ctggctgaac agggctatga agtggcacaa agcaatgcag cctttattct   1440

tgatcagaga gaagcaagca ttgtaggtga gaatgaaact tatcccagag ctttgctaca   1500

ttggaacagg gccgcctctc aaggctatac tgtggctaga attaagctcg gagactacca   1560

tttctatggg tttggcaccg atgtagatta tgaaactgca tttattcatt accgtctggc   1620

ttctgagcag caacacagtg cacaagctat gtttaatctg ggatatatgc atgagaaagg   1680

actgggcatt aaacaggata ttcaccttgc gaaacgtttt tatgacatgg cagctgaagc   1740

cagcccagat gcacaagttc cagtcttcct agccctctgc aaattgggcg tcgtctattt   1800

cttgcagtac atacgggaaa caaacattcg agatatgttc acccaacttg atatggacca   1860

gcttttggga cctgagtggg acctttacct catgaccatc attgcgctgc tgttgggaac   1920

agtcatagct tacaggcaaa ggcagcacca agacatgcct gcacccaggc ctccagggcc   1980

acggccagct ccaccccagc aggaggggcc accagagcag cagccaccac agtaa        2035


<210>  3
<211>  650
<212>  PRT
<213>  Artificial

<220>
<223>  scFv9E1-degradin protein

<400>  3

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


Val His Ser Gln Ile Val Leu Thr Gln Ser Pro Ala Ile Met Ser Ala 
            20                  25                  30          


Ser Pro Gly Glu Arg Val Thr Ile Thr Cys Ser Val Ser Ser Thr Val 
        35                  40                  45              


Ser Tyr Met His Trp Phe Gln Gln Lys Pro Gly Thr Ser Pro Lys Val 
    50                  55                  60                  


Leu Ile His Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Ala Arg Phe 
65                  70                  75                  80  


Ser Gly Ser Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Arg Met 
                85                  90                  95      


Glu Ala Glu Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Arg Ser Tyr Phe 
            100                 105                 110         


Pro Leu Thr Phe Gly Ile Gly Thr Lys Leu Glu Leu Lys Gly Ser Thr 
        115                 120                 125             


Ser Gly Ser Gly Lys Pro Gly Ser Gly Glu Gly Ser Ser Lys Gly Gln 
    130                 135                 140                 


Val Gln Leu Lys Glu Ser Gly Pro Gly Leu Val Ala Pro Ser Gln Ser 
145                 150                 155                 160 


Leu Ser Ile Thr Cys Thr Val Ser Gly Phe Ser Leu Thr Ser Tyr Gly 
                165                 170                 175     


Ile His Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Leu Gly 
            180                 185                 190         


Val Ile Trp Ala Gly Gly Asn Thr Ser Tyr Asn Ser Ala Leu Met Ser 
        195                 200                 205             


Arg Leu Ser Ile Ser Lys Asp Asn Ser Arg Ser Gln Val Ile Leu Gln 
    210                 215                 220                 


Met Asn Ser Leu Gln Thr Asp Asp Thr Ala Ile Tyr Tyr Cys Ala Arg 
225                 230                 235                 240 


Glu Asp Arg Met Tyr Trp Tyr Phe Asp Val Trp Gly Ala Gly Thr Thr 
                245                 250                 255     


Val Thr Val Ser Ser Gly Ala Ser Gly Lys Pro Ile Pro Asn Pro Leu 
            260                 265                 270         


Leu Gly Leu Asp Ala Ser Gly Ser Asp Ile Val Pro Gln Ser Asn Glu 
        275                 280                 285             


Thr Ala Leu His Tyr Phe Lys Lys Ala Ala Asp Met Gly Asn Pro Val 
    290                 295                 300                 


Gly Gln Ser Gly Leu Gly Met Ala Tyr Leu Tyr Gly Arg Gly Val Gln 
305                 310                 315                 320 


Val Asn Tyr Asp Leu Ala Leu Lys Tyr Phe Gln Lys Ala Ala Glu Gln 
                325                 330                 335     


Gly Trp Val Asp Gly Gln Leu Gln Leu Gly Ser Met Tyr Tyr Asn Gly 
            340                 345                 350         


Ile Gly Val Lys Arg Asp Tyr Lys Gln Ala Leu Lys Tyr Phe Asn Leu 
        355                 360                 365             


Ala Ser Gln Gly Gly His Ile Leu Ala Phe Tyr Asn Leu Ala Gln Met 
    370                 375                 380                 


His Ala Ser Gly Thr Gly Val Met Arg Ser Cys His Thr Ala Val Glu 
385                 390                 395                 400 


Leu Phe Lys Asn Val Cys Glu Arg Gly Arg Trp Ser Glu Arg Leu Met 
                405                 410                 415     


Thr Ala Tyr Asn Ser Tyr Lys Asp Gly Asp Tyr Asn Ala Ala Val Ile 
            420                 425                 430         


Gln Tyr Leu Leu Leu Ala Glu Gln Gly Tyr Glu Val Ala Gln Ser Asn 
        435                 440                 445             


Ala Ala Phe Ile Leu Asp Gln Arg Glu Ala Ser Ile Val Gly Glu Asn 
    450                 455                 460                 


Glu Thr Tyr Pro Arg Ala Leu Leu His Trp Asn Arg Ala Ala Ser Gln 
465                 470                 475                 480 


Gly Tyr Thr Val Ala Arg Ile Lys Leu Gly Asp Tyr His Phe Tyr Gly 
                485                 490                 495     


Phe Gly Thr Asp Val Asp Tyr Glu Thr Ala Phe Ile His Tyr Arg Leu 
            500                 505                 510         


Ala Ser Glu Gln Gln His Ser Ala Gln Ala Met Phe Asn Leu Gly Tyr 
        515                 520                 525             


Met His Glu Lys Gly Leu Gly Ile Lys Gln Asp Ile His Leu Ala Lys 
    530                 535                 540                 


Arg Phe Tyr Asp Met Ala Ala Glu Ala Ser Pro Asp Ala Gln Val Pro 
545                 550                 555                 560 


Val Phe Leu Ala Leu Cys Lys Leu Gly Val Val Tyr Phe Leu Gln Tyr 
                565                 570                 575     


Ile Arg Glu Thr Asn Ile Arg Asp Met Phe Thr Gln Leu Asp Met Asp 
            580                 585                 590         


Gln Leu Leu Gly Pro Glu Trp Asp Leu Tyr Leu Met Thr Ile Ile Ala 
        595                 600                 605             


Leu Leu Leu Gly Thr Val Ile Ala Tyr Arg Gln Arg Gln His Gln Asp 
    610                 615                 620                 


Met Pro Ala Pro Arg Pro Pro Gly Pro Arg Pro Ala Pro Pro Gln Gln 
625                 630                 635                 640 


Glu Gly Pro Pro Glu Gln Gln Pro Pro Gln 
                645                 650 


<210>  4
<211>  1969
<212>  DNA
<213>  Artificial

<220>
<223>  LigandFcERI-degradin-gene

<400>  4
atgggctgga gcctgatcct cctgttcctc gtcgctgtgg ctacaggtaa ggggctcaca     60

gtagcaggct tgaggtctgg acatatatat gggtgacaat gacatccact ttgcctttct    120

ctccacaggt gtgcactcgg aaggcaaacc aatcccaaac ccactgctgg gcctggatag    180

ttgtgcagat tcgaacccga gaggggtgag cgcctaccta agccggccca gcccgttcga    240

cctgttcatc cgcaagtcgc ccacgatcac ctgtctggtg gtggacctgg cacccagcaa    300

ggggaccgtg aacctgacct ggtcccgggc cagtgggaag cctgtgaacc actccaccag    360

aaaggaggag aagcagcgca atggcacgtt aaccgtcacg tccaccctgc cggtgggcac    420

ccgagactgg atcgaggggg agacctacca gtgcagggtg acccaccccc acctgcccag    480

ggccctcatg cggtccacga ccaagaccag cggcccgcgt gctgccccgg aagtctatgc    540

gtttgcgacg ccggagtggc cggggagccg ggacaagcgc accctcgcct gcctgatcca    600

gaacttcatg cctgaggaca tctcggtgca gtggctgcac aacgaggtgc agctcccgga    660

cgcccggcac agcacgacgc agccccgcaa gaccaagggc tccggcttct tcgtcttcag    720

ccgcctggag gtgaccaggg ccgaatggga gcagaaagat gagttcatct gccgtgcagt    780

ccatgaggca gcgagcccct cacagaccgt ccagcgagcg gtgtctgtaa atcccggggg    840

atccgctagc ggaagtgaca ttgtacctca gagtaatgag acagctctcc actactttaa    900

gaaagctgct gacatgggca acccagttgg acagagtggg cttggaatgg cctacctcta    960

tgggagagga gttcaagtta attatgatct agcccttaag tatttccaga aagctgctga   1020

acaaggctgg gtggatgggc agctacagct tggttccatg tactataatg gcattggagt   1080

caagagagat tataaacagg ccttgaagta ttttaattta gcttctcagg gaggccatat   1140

cttggctttc tataacctag ctcagatgca tgccagtggc accggcgtga tgcgatcatg   1200

tcacactgca gtggagttgt ttaagaatgt atgtgaacga ggccgttggt ctgaaaggct   1260

tatgactgcc tataacagct ataaagatgg cgattacaat gctgcagtga tccagtacct   1320

cctcctggct gaacagggct atgaagtggc acaaagcaat gcagccttta ttcttgatca   1380

gagagaagca agcattgtag gtgagaatga aacttatccc agagctttgc tacattggaa   1440

cagggccgcc tctcaaggct atactgtggc tagaattaag ctcggagact accatttcta   1500

tgggtttggc accgatgtag attatgaaac tgcatttatt cattaccgtc tggcttctga   1560

gcagcaacac agtgcacaag ctatgtttaa tctgggatat atgcatgaga aaggactggg   1620

cattaaacag gatattcacc ttgcgaaacg tttttatgac atggcagctg aagccagccc   1680

agatgcacaa gttccagtct tcctagccct ctgcaaattg ggcgtcgtct atttcttgca   1740

gtacatacgg gaaacaaaca ttcgagatat gttcacccaa cttgatatgg accagctttt   1800

gggacctgag tgggaccttt acctcatgac catcattgcg ctgctgttgg gaacagtcat   1860

agcttacagg caaaggcagc accaagacat gcctgcaccc aggcctccag ggccacggcc   1920

agctccaccc cagcaggagg ggccaccaga gcagcagcca ccacagtaa               1969


<210>  5
<211>  618
<212>  PRT
<213>  Artificial

<220>
<223>  LigandFcERI-degradin protein

<400>  5

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


Val His Ser Glu Gly Lys Pro Ile Pro Asn Pro Leu Leu Gly Leu Asp 
            20                  25                  30          


Ser Cys Ala Asp Ser Asn Pro Arg Gly Val Ser Ala Tyr Leu Ser Arg 
        35                  40                  45              


Pro Ser Pro Phe Asp Leu Phe Ile Arg Lys Ser Pro Thr Ile Thr Cys 
    50                  55                  60                  


Leu Val Val Asp Leu Ala Pro Ser Lys Gly Thr Val Asn Leu Thr Trp 
65                  70                  75                  80  


Ser Arg Ala Ser Gly Lys Pro Val Asn His Ser Thr Arg Lys Glu Glu 
                85                  90                  95      


Lys Gln Arg Asn Gly Thr Leu Thr Val Thr Ser Thr Leu Pro Val Gly 
            100                 105                 110         


Thr Arg Asp Trp Ile Glu Gly His Leu Pro Arg Ala Leu Met Arg Ser 
        115                 120                 125             


Thr Thr Lys Thr Ser Gly Pro Arg Ala Ala Pro Glu Val Tyr Ala Phe 
    130                 135                 140                 


Ala Thr Pro Glu Trp Pro Gly Ser Arg Asp Lys Arg Thr Leu Ala Cys 
145                 150                 155                 160 


Leu Ile Gln Asn Phe Met Pro Glu Asp Ile Ser Val Gln Trp Leu His 
                165                 170                 175     


Asn Glu Val Gln Leu Pro Asp Ala Arg His Ser Thr Thr Gln Pro Arg 
            180                 185                 190         


Lys Thr Lys Gly Ser Gly Phe Phe Val Phe Ser Arg Leu Glu Val Thr 
        195                 200                 205             


Arg Ala Glu Trp Glu Gln Lys Asp Glu Phe Ile Cys Arg Ala Val His 
    210                 215                 220                 


Glu Ala Ala Ser Pro Ser Gln Thr Val Gln Arg Ala Val Ser Val Asn 
225                 230                 235                 240 


Pro Gly Gly Ser Ala Ser Gly Ser Asp Ile Val Pro Gln Ser Asn Glu 
                245                 250                 255     


Thr Ala Leu His Tyr Phe Lys Lys Ala Ala Asp Met Gly Asn Pro Val 
            260                 265                 270         


Gly Gln Ser Gly Leu Gly Met Ala Tyr Leu Tyr Gly Arg Gly Val Gln 
        275                 280                 285             


Val Asn Tyr Asp Leu Ala Leu Lys Tyr Phe Gln Lys Ala Ala Glu Gln 
    290                 295                 300                 


Gly Trp Val Asp Gly Gln Leu Gln Leu Gly Ser Met Tyr Tyr Asn Gly 
305                 310                 315                 320 


Ile Gly Val Lys Arg Asp Tyr Lys Gln Ala Leu Lys Tyr Phe Asn Leu 
                325                 330                 335     


Ala Ser Gln Gly Gly His Ile Leu Ala Phe Tyr Asn Leu Ala Gln Met 
            340                 345                 350         


His Ala Ser Gly Thr Gly Val Met Arg Ser Cys His Thr Ala Val Glu 
        355                 360                 365             


Leu Phe Lys Asn Val Cys Glu Arg Gly Arg Trp Ser Glu Arg Leu Met 
    370                 375                 380                 


Thr Ala Tyr Asn Ser Tyr Lys Asp Gly Asp Tyr Asn Ala Ala Val Ile 
385                 390                 395                 400 


Gln Tyr Leu Leu Leu Ala Glu Gln Gly Tyr Glu Val Ala Gln Ser Asn 
                405                 410                 415     


Ala Ala Phe Ile Leu Asp Gln Arg Glu Ala Ser Ile Val Gly Glu Asn 
            420                 425                 430         


Glu Thr Tyr Pro Arg Ala Leu Leu His Trp Asn Arg Ala Ala Ser Gln 
        435                 440                 445             


Gly Tyr Thr Val Ala Arg Ile Lys Leu Gly Asp Tyr His Phe Tyr Gly 
    450                 455                 460                 


Phe Gly Thr Asp Val Asp Tyr Glu Thr Ala Phe Ile His Tyr Arg Leu 
465                 470                 475                 480 


Ala Ser Glu Gln Gln His Ser Ala Gln Ala Met Phe Asn Leu Gly Tyr 
                485                 490                 495     


Met His Glu Lys Gly Leu Gly Ile Lys Gln Asp Ile His Leu Ala Lys 
            500                 505                 510         


Arg Phe Tyr Asp Met Ala Ala Glu Ala Ser Pro Asp Ala Gln Val Pro 
        515                 520                 525             


Val Phe Leu Ala Leu Cys Lys Leu Gly Val Val Tyr Phe Leu Gln Tyr 
    530                 535                 540                 


Ile Arg Glu Thr Asn Ile Arg Asp Met Phe Thr Gln Leu Asp Met Asp 
545                 550                 555                 560 


Gln Leu Leu Gly Pro Glu Trp Asp Leu Tyr Leu Met Thr Ile Ile Ala 
                565                 570                 575     


Leu Leu Leu Gly Thr Val Ile Ala Tyr Arg Gln Arg Gln His Gln Asp 
            580                 585                 590         


Met Pro Ala Pro Arg Pro Pro Gly Pro Arg Pro Ala Pro Pro Gln Gln 
        595                 600                 605             


Glu Gly Pro Pro Glu Gln Gln Pro Pro Gln 
    610                 615             


<210>  6
<211>  30
<212>  DNA
<213>  Artificial

<220>
<223>  primer SEL1L-NheI

<400>  6
agtagctagc ggaagtgaca ttgtacctca                                      30


<210>  7
<211>  31
<212>  DNA
<213>  Artificial

<220>
<223>  primer SEL1L-EcoRI

<400>  7
tcagaattct tactgtggtg gctgctgctc t                                    31


<210>  8
<211>  48
<212>  DNA
<213>  Artificial

<220>
<223>  linker encoding the SV5 tag

<400>  8
gctagtggca aaccaatccc aaacccactg ctgggcctgg atgctagc                  48


<210>  9
<211>  31
<212>  DNA
<213>  Artificial

<220>
<223>  primer SEL1L Delta-NheI

<400>  9
agtagctagc gatatggacc agcttttggg a                                    31


<210>  10
<211>  69
<212>  DNA
<213>  Artificial

<220>
<223>  linker encoding the SV5 tag followed by the KDEL sequence

<400>  10
tccggaggca aaccaatccc aaacccactg ctgggcctgg atagtactaa agatgagctg     60

tagctcgag                                                             69


