               SEQUENCE LISTING

<110> B.R.A.I.N. Biotechnology Research And Information Network AG

<120> OPTIMIZATION OF THE EXPRESSION OF SERINE PROTEASES IN HOST CELLS

<130> AA1177 PCT

<150> EP17 17 3813.1
<151> 2017-05-31

<160> 12

<170> BiSSAP 1.3

<210> 1
<211> 58
<212> PRT
<213> Bos taurus

<400> 1
Arg Pro Asp Phe Cys Leu Glu Pro Pro Tyr Thr Gly Pro Cys Lys Ala 
1               5                   10                  15      
Arg Ile Ile Arg Tyr Phe Tyr Asn Ala Lys Ala Gly Leu Cys Gln Thr 
            20                  25                  30          
Phe Val Tyr Gly Gly Cys Arg Ala Lys Arg Asn Asn Phe Lys Ser Ala 
        35                  40                  45              
Glu Asp Cys Met Arg Thr Cys Gly Gly Ala 
    50                  55              

<210> 2
<211> 177
<212> DNA
<213> Artificial Sequence

<220> 
<223> Codon-optimized nucleotide sequence of aprotinin

<400> 2
agaccagact tctgtttgga accaccatac actggtccat gtaaggctag aatcatcaga     60

tacttctaca acgctaaggc tggtttgtgt cagactttcg tttacggtgg ttgtagagct    120

aagagaaaca acttcaagtc cgctgaggac tgtatgagaa cttgtggtgg tgcttaa       177


<210> 3
<211> 254
<212> PRT
<213> Lucilia sericata

<400> 3
Met Phe Arg Phe Val Ala Leu Phe Ala Phe Val Ser Cys Ala Leu Ala 
1               5                   10                  15      
Gly Ala Ile Pro Asn Asp Leu Asp Gly Arg Ile Val Asn Gly Val Asp 
            20                  25                  30          
Thr Thr Ile Gln Ala His Pro Tyr Gln Val Ser Leu Gln Thr Asn Asn 
        35                  40                  45              
Gly Phe His Phe Cys Gly Gly Ser Ile Ile Ser Glu Asp Ile Ile Val 
    50                  55                  60                  
Thr Ala Ala His Cys Met Gln Ser Tyr Lys Ala Tyr Gln Phe Lys Val 
65                  70                  75                  80  
Arg Leu Gly Ser Thr Glu Tyr Asp Asn Gly Gly Glu Leu Val Ala Val 
                85                  90                  95      
Lys Ser Phe Lys Tyr His Glu Gly Tyr Asn Pro Glu Thr Met Val Asn 
            100                 105                 110         
Asp Val Ala Val Ile Lys Leu Ala Thr Pro Val Arg Glu Ser Ser Lys 
        115                 120                 125             
Val Arg Tyr Val Lys Leu Ala Glu Lys Thr Pro Ala Thr Gly Thr Pro 
    130                 135                 140                 
Ala Val Val Thr Gly Trp Gly Ser Lys Cys Phe Leu Phe Cys Gln Thr 
145                 150                 155                 160 
Ala Pro Lys Val Leu Gln Lys Val Glu Val Asp Ile Val Asp Glu Lys 
                165                 170                 175     
Thr Cys Ala Ser Ser Glu Tyr Lys Tyr Gly Asp Asp Ile Lys Glu Thr 
            180                 185                 190         
Met Leu Cys Ala Tyr Ala Val Lys Lys Asp Ala Cys Gln Gly Asp Ser 
        195                 200                 205             
Gly Gly Pro Leu Val Ala Asn Asn Lys Leu Val Gly Val Val Ser Trp 
    210                 215                 220                 
Gly Lys Gly Cys Ala Leu Ala Gly Tyr Pro Gly Val Tyr Cys Asp Val 
225                 230                 235                 240 
Ala Thr Val Arg Ser Trp Ile Glu Lys Thr Ala Lys Ser Leu 
                245                 250                 

<210> 4
<211> 238
<212> PRT
<213> Lucilia sericata

<400> 4
Gly Ala Ile Pro Asn Asp Leu Asp Gly Arg Ile Val Asn Gly Val Asp 
1               5                   10                  15      
Thr Thr Ile Gln Ala His Pro Tyr Gln Val Ser Leu Gln Thr Asn Asn 
            20                  25                  30          
Gly Phe His Phe Cys Gly Gly Ser Ile Ile Ser Glu Asp Ile Ile Val 
        35                  40                  45              
Thr Ala Ala His Cys Met Gln Ser Tyr Lys Ala Tyr Gln Phe Lys Val 
    50                  55                  60                  
Arg Leu Gly Ser Thr Glu Tyr Asp Asn Gly Gly Glu Leu Val Ala Val 
65                  70                  75                  80  
Lys Ser Phe Lys Tyr His Glu Gly Tyr Asn Pro Glu Thr Met Val Asn 
                85                  90                  95      
Asp Val Ala Val Ile Lys Leu Ala Thr Pro Val Arg Glu Ser Ser Lys 
            100                 105                 110         
Val Arg Tyr Val Lys Leu Ala Glu Lys Thr Pro Ala Thr Gly Thr Pro 
        115                 120                 125             
Ala Val Val Thr Gly Trp Gly Ser Lys Cys Phe Leu Phe Cys Gln Thr 
    130                 135                 140                 
Ala Pro Lys Val Leu Gln Lys Val Glu Val Asp Ile Val Asp Glu Lys 
145                 150                 155                 160 
Thr Cys Ala Ser Ser Glu Tyr Lys Tyr Gly Asp Asp Ile Lys Glu Thr 
                165                 170                 175     
Met Leu Cys Ala Tyr Ala Val Lys Lys Asp Ala Cys Gln Gly Asp Ser 
            180                 185                 190         
Gly Gly Pro Leu Val Ala Asn Asn Lys Leu Val Gly Val Val Ser Trp 
        195                 200                 205             
Gly Lys Gly Cys Ala Leu Ala Gly Tyr Pro Gly Val Tyr Cys Asp Val 
    210                 215                 220                 
Ala Thr Val Arg Ser Trp Ile Glu Lys Thr Ala Lys Ser Leu 
225                 230                 235             

<210> 5
<211> 228
<212> PRT
<213> Lucilia sericata

<400> 5
Ile Val Asn Gly Val Asp Thr Thr Ile Gln Ala His Pro Tyr Gln Val 
1               5                   10                  15      
Ser Leu Gln Thr Asn Asn Gly Phe His Phe Cys Gly Gly Ser Ile Ile 
            20                  25                  30          
Ser Glu Asp Ile Ile Val Thr Ala Ala His Cys Met Gln Ser Tyr Lys 
        35                  40                  45              
Ala Tyr Gln Phe Lys Val Arg Leu Gly Ser Thr Glu Tyr Asp Asn Gly 
    50                  55                  60                  
Gly Glu Leu Val Ala Val Lys Ser Phe Lys Tyr His Glu Gly Tyr Asn 
65                  70                  75                  80  
Pro Glu Thr Met Val Asn Asp Val Ala Val Ile Lys Leu Ala Thr Pro 
                85                  90                  95      
Val Arg Glu Ser Ser Lys Val Arg Tyr Val Lys Leu Ala Glu Lys Thr 
            100                 105                 110         
Pro Ala Thr Gly Thr Pro Ala Val Val Thr Gly Trp Gly Ser Lys Cys 
        115                 120                 125             
Phe Leu Phe Cys Gln Thr Ala Pro Lys Val Leu Gln Lys Val Glu Val 
    130                 135                 140                 
Asp Ile Val Asp Glu Lys Thr Cys Ala Ser Ser Glu Tyr Lys Tyr Gly 
145                 150                 155                 160 
Asp Asp Ile Lys Glu Thr Met Leu Cys Ala Tyr Ala Val Lys Lys Asp 
                165                 170                 175     
Ala Cys Gln Gly Asp Ser Gly Gly Pro Leu Val Ala Asn Asn Lys Leu 
            180                 185                 190         
Val Gly Val Val Ser Trp Gly Lys Gly Cys Ala Leu Ala Gly Tyr Pro 
        195                 200                 205             
Gly Val Tyr Cys Asp Val Ala Thr Val Arg Ser Trp Ile Glu Lys Thr 
    210                 215                 220                 
Ala Lys Ser Leu 
225             

<210> 6
<211> 765
<212> DNA
<213> Lucilia sericata

<400> 6
atgttccggt ttgtagctct attcgctttc gttagctgtg ccttggcggg cgctattccc      60

aatgatttgg atggccgcat tgtcaatggt gtggatacca ccattcaggc ccatccctat     120

caggtttctt tgcaaaccaa caatggtttc catttctgcg gtggttccat catcagcgaa     180

gacattattg taactgctgc tcattgcatg caatcctaca aggcctacca attcaaagta     240

cgtttgggtt ccactgaata cgataatggt ggtgaattgg ttgccgtcaa gtctttcaaa     300

taccacgaag gttacaatcc cgaaaccatg gttaatgatg ttgccgttat caaattagcc     360

actccagtgc gtgaatcttc caaggtacgt tatgttaaat tggctgagaa gacacctgct     420

actggcaccc cagctgtcgt tactggttgg ggttctaagt gcttcttgtt ctgccaaact     480

gcccctaaag ttttgcaaaa ggttgaggtc gatattgttg atgagaagac ctgcgcttcc     540

agcgaataca aatatggtga tgacatcaag gaaactatgt tgtgtgctta tgctgttaag     600

aaggatgctt gccaaggtga ttctggtggt cctttggttg ccaacaacaa attggtcggt     660

gttgtttcct ggggtaaagg ttgtgccctt gctggctatc ccggtgtata ctgcgatgtt     720

gctactgtcc gcagctggat tgaaaagact gccaagagtt tgtaa                     765


<210> 7
<211> 717
<212> DNA
<213> Lucilia sericata

<400> 7
ggcgctattc ccaatgattt ggatggccgc attgtcaatg gtgtggatac caccattcag      60

gcccatccct atcaggtttc tttgcaaacc aacaatggtt tccatttctg cggtggttcc     120

atcatcagcg aagacattat tgtaactgct gctcattgca tgcaatccta caaggcctac     180

caattcaaag tacgtttggg ttccactgaa tacgataatg gtggtgaatt ggttgccgtc     240

aagtctttca aataccacga aggttacaat cccgaaacca tggttaatga tgttgccgtt     300

atcaaattag ccactccagt gcgtgaatct tccaaggtac gttatgttaa attggctgag     360

aagacacctg ctactggcac cccagctgtc gttactggtt ggggttctaa gtgcttcttg     420

ttctgccaaa ctgcccctaa agttttgcaa aaggttgagg tcgatattgt tgatgagaag     480

acctgcgctt ccagcgaata caaatatggt gatgacatca aggaaactat gttgtgtgct     540

tatgctgtta agaaggatgc ttgccaaggt gattctggtg gtcctttggt tgccaacaac     600

aaattggtcg gtgttgtttc ctggggtaaa ggttgtgccc ttgctggcta tcccggtgta     660

tactgcgatg ttgctactgt ccgcagctgg attgaaaaga ctgccaagag tttgtaa        717


<210> 8
<211> 687
<212> DNA
<213> Lucilia sericata

<400> 8
attgtcaatg gtgtggatac caccattcag gcccatccct atcaggtttc tttgcaaacc      60

aacaatggtt tccatttctg cggtggttcc atcatcagcg aagacattat tgtaactgct     120

gctcattgca tgcaatccta caaggcctac caattcaaag tacgtttggg ttccactgaa     180

tacgataatg gtggtgaatt ggttgccgtc aagtctttca aataccacga aggttacaat     240

cccgaaacca tggttaatga tgttgccgtt atcaaattag ccactccagt gcgtgaatct     300

tccaaggtac gttatgttaa attggctgag aagacacctg ctactggcac cccagctgtc     360

gttactggtt ggggttctaa gtgcttcttg ttctgccaaa ctgcccctaa agttttgcaa     420

aaggttgagg tcgatattgt tgatgagaag acctgcgctt ccagcgaata caaatatggt     480

gatgacatca aggaaactat gttgtgtgct tatgctgtta agaaggatgc ttgccaaggt     540

gattctggtg gtcctttggt tgccaacaac aaattggtcg gtgttgtttc ctggggtaaa     600

ggttgtgccc ttgctggcta tcccggtgta tactgcgatg ttgctactgt ccgcagctgg     660

attgaaaaga ctgccaagag tttgtaa                                         687


<210> 9
<211> 60
<212> PRT
<213> Artificial Sequence

<220> 
<223> Aprotinin fused to EA-Tag

<400> 9
Glu Ala Arg Pro Asp Phe Cys Leu Glu Pro Pro Tyr Thr Gly Pro Cys 
1               5                   10                  15      
Lys Ala Arg Ile Ile Arg Tyr Phe Tyr Asn Ala Lys Ala Gly Leu Cys 
            20                  25                  30          
Gln Thr Phe Val Tyr Gly Gly Cys Arg Ala Lys Arg Asn Asn Phe Lys 
        35                  40                  45              
Ser Ala Glu Asp Cys Met Arg Thr Cys Gly Gly Ala 
    50                  55                  60  

<210> 10
<211> 183
<212> DNA
<213> Artificial Sequence

<220> 
<223> Aprotinin fused to GAAGCT-Tag

<400> 10
gaagctagac cagacttctg tttggaacca ccatacactg gtccatgtaa ggctagaatc     60

atcagatact tctacaacgc taaggctggt ttgtgtcaga ctttcgttta cggtggttgt    120

agagctaaga gaaacaactt caagtccgct gaggactgta tgagaacttg tggtggtgct    180

taa                                                                  183


<210> 11
<211> 438
<212> DNA
<213> Artificial Sequence

<220> 
<223> Aprotinin sequence fused to sequence of the alpha mating factor
      from S. cerevisiae

<400> 11
atgagattcc catctatttt caccgctgtc ttgttcgctg cctcctctgc attggctgcc      60

cctgttaaca ctaccactga agacgagact gctcaaattc cagctgaagc agttatcggt     120

tactctgacc ttgagggtga tttcgacgtc gctgttttgc ctttctctaa ctccactaac     180

aacggtttgt tgttcattaa caccactatc gcttccattg ctgctaagga agagggtgtc     240

tctctcgaga aaagagaagc tagaccagac ttctgtttgg aaccaccata cactggtcca     300

tgtaaggcta gaatcatcag atacttctac aacgctaagg ctggtttgtg tcagactttc     360

gtttacggtg gttgtagagc taagagaaac aacttcaagt ccgctgagga ctgtatgaga     420

acttgtggtg gtgcttaa                                                   438


<210> 12
<211> 145
<212> PRT
<213> Artificial Sequence

<220> 
<223> Aprotinin sequence fused to sequence of the alpha mating factor
      from S. cerevisiae

<400> 12
Met Arg Phe Pro Ser Ile Phe Thr Ala Val Leu Phe Ala Ala Ser Ser 
1               5                   10                  15      
Ala Leu Ala Ala Pro Val Asn Thr Thr Thr Glu Asp Glu Thr Ala Gln 
            20                  25                  30          
Ile Pro Ala Glu Ala Val Ile Gly Tyr Ser Asp Leu Glu Gly Asp Phe 
        35                  40                  45              
Asp Val Ala Val Leu Pro Phe Ser Asn Ser Thr Asn Asn Gly Leu Leu 
    50                  55                  60                  
Phe Ile Asn Thr Thr Ile Ala Ser Ile Ala Ala Lys Glu Glu Gly Val 
65                  70                  75                  80  
Ser Leu Glu Lys Arg Glu Ala Arg Pro Asp Phe Cys Leu Glu Pro Pro 
                85                  90                  95      
Tyr Thr Gly Pro Cys Lys Ala Arg Ile Ile Arg Tyr Phe Tyr Asn Ala 
            100                 105                 110         
Lys Ala Gly Leu Cys Gln Thr Phe Val Tyr Gly Gly Cys Arg Ala Lys 
        115                 120                 125             
Arg Asn Asn Phe Lys Ser Ala Glu Asp Cys Met Arg Thr Cys Gly Gly 
    130                 135                 140                 
Ala 
145 

