                         SEQUENCE LISTING

<110>  VIB VZW
       VRIJE UNIVERSITEIT BRUSSEL
 
<120>  Secretion and functional display of chimeric polypeptides

<130>  HR/CURLI/484

<150>  EP 13199513.6
<151>  2013-12-24

<160>  77    

<170>  PatentIn version 3.5

<210>  1
<211>  151
<212>  PRT
<213>  Escherichia coli

<400>  1

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


Ser Ala Leu Ala Gly Val Val Pro Gln Tyr Gly Gly Gly Gly Asn His 
            20                  25                  30          


Gly Gly Gly Gly Asn Asn Ser Gly Pro Asn Ser Glu Leu Asn Ile Tyr 
        35                  40                  45              


Gln Tyr Gly Gly Gly Asn Ser Ala Leu Ala Leu Gln Thr Asp Ala Arg 
    50                  55                  60                  


Asn Ser Asp Leu Thr Ile Thr Gln His Gly Gly Gly Asn Gly Ala Asp 
65                  70                  75                  80  


Val Gly Gln Gly Ser Asp Asp Ser Ser Ile Asp Leu Thr Gln Arg Gly 
                85                  90                  95      


Phe Gly Asn Ser Ala Thr Leu Asp Gln Trp Asn Gly Lys Asn Ser Glu 
            100                 105                 110         


Met Thr Val Lys Gln Phe Gly Gly Gly Asn Gly Ala Ala Val Asp Gln 
        115                 120                 125             


Thr Ala Ser Asn Ser Ser Val Asn Val Thr Gln Val Gly Phe Gly Asn 
    130                 135                 140                 


Asn Ala Thr Ala His Gln Tyr 
145                 150     


<210>  2
<211>  131
<212>  PRT
<213>  Escherichia coli

<400>  2

Gly Val Val Pro Gln Tyr Gly Gly Gly Gly Asn His Gly Gly Gly Gly 
1               5                   10                  15      


Asn Asn Ser Gly Pro Asn Ser Glu Leu Asn Ile Tyr Gln Tyr Gly Gly 
            20                  25                  30          


Gly Asn Ser Ala Leu Ala Leu Gln Thr Asp Ala Arg Asn Ser Asp Leu 
        35                  40                  45              


Thr Ile Thr Gln His Gly Gly Gly Asn Gly Ala Asp Val Gly Gln Gly 
    50                  55                  60                  


Ser Asp Asp Ser Ser Ile Asp Leu Thr Gln Arg Gly Phe Gly Asn Ser 
65                  70                  75                  80  


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


Gln Phe Gly Gly Gly Asn Gly Ala Ala Val Asp Gln Thr Ala Ser Asn 
            100                 105                 110         


Ser Ser Val Asn Val Thr Gln Val Gly Phe Gly Asn Asn Ala Thr Ala 
        115                 120                 125             


His Gln Tyr 
    130     


<210>  3
<211>  109
<212>  PRT
<213>  Escherichia coli

<400>  3

Ser Glu Leu Asn Ile Tyr Gln Tyr Gly Gly Gly Asn Ser Ala Leu Ala 
1               5                   10                  15      


Leu Gln Thr Asp Ala Arg Asn Ser Asp Leu Thr Ile Thr Gln His Gly 
            20                  25                  30          


Gly Gly Asn Gly Ala Asp Val Gly Gln Gly Ser Asp Asp Ser Ser Ile 
        35                  40                  45              


Asp Leu Thr Gln Arg Gly Phe Gly Asn Ser Ala Thr Leu Asp Gln Trp 
    50                  55                  60                  


Asn Gly Lys Asn Ser Glu Met Thr Val Lys Gln Phe Gly Gly Gly Asn 
65                  70                  75                  80  


Gly Ala Ala Val Asp Gln Thr Ala Ser Asn Ser Ser Val Asn Val Thr 
                85                  90                  95      


Gln Val Gly Phe Gly Asn Asn Ala Thr Ala His Gln Tyr 
            100                 105                 


<210>  4
<211>  23
<212>  PRT
<213>  Escherichia coli

<400>  4

Ser Glu Leu Asn Ile Tyr Gln Tyr Gly Gly Gly Asn Ser Ala Leu Ala 
1               5                   10                  15      


Leu Gln Thr Asp Ala Arg Asn 
            20              


<210>  5
<211>  22
<212>  PRT
<213>  Escherichia coli

<400>  5

Ser Asp Leu Thr Ile Thr Gln His Gly Gly Gly Asn Gly Ala Asp Val 
1               5                   10                  15      


Gly Gln Gly Ser Asp Asp 
            20          


<210>  6
<211>  23
<212>  PRT
<213>  Escherichia coli

<400>  6

Ser Ser Ile Asp Leu Thr Gln Arg Gly Phe Gly Asn Ser Ala Thr Leu 
1               5                   10                  15      


Asp Gln Trp Asn Gly Lys Asn 
            20              


<210>  7
<211>  22
<212>  PRT
<213>  Escherichia coli

<400>  7

Ser Glu Met Thr Val Lys Gln Phe Gly Gly Gly Asn Gly Ala Ala Val 
1               5                   10                  15      


Asp Gln Thr Ala Ser Asn 
            20          


<210>  8
<211>  19
<212>  PRT
<213>  Escherichia coli

<400>  8

Ser Ser Val Asn Val Thr Gln Val Gly Phe Gly Asn Asn Ala Thr Ala 
1               5                   10                  15      


His Gln Tyr 
            


<210>  9
<211>  45
<212>  PRT
<213>  Escherichia coli

<400>  9

Ser Glu Leu Asn Ile Tyr Gln Tyr Gly Gly Gly Asn Ser Ala Leu Ala 
1               5                   10                  15      


Leu Gln Thr Asp Ala Arg Asn Ser Asp Leu Thr Ile Thr Gln His Gly 
            20                  25                  30          


Gly Gly Asn Gly Ala Asp Val Gly Gln Gly Ser Asp Asp 
        35                  40                  45  


<210>  10
<211>  68
<212>  PRT
<213>  Escherichia coli

<400>  10

Ser Glu Leu Asn Ile Tyr Gln Tyr Gly Gly Gly Asn Ser Ala Leu Ala 
1               5                   10                  15      


Leu Gln Thr Asp Ala Arg Asn Ser Asp Leu Thr Ile Thr Gln His Gly 
            20                  25                  30          


Gly Gly Asn Gly Ala Asp Val Gly Gln Gly Ser Asp Asp Ser Ser Ile 
        35                  40                  45              


Asp Leu Thr Gln Arg Gly Phe Gly Asn Ser Ala Thr Leu Asp Gln Trp 
    50                  55                  60                  


Asn Gly Lys Asn 
65              


<210>  11
<211>  90
<212>  PRT
<213>  Escherichia coli

<400>  11

Ser Glu Leu Asn Ile Tyr Gln Tyr Gly Gly Gly Asn Ser Ala Leu Ala 
1               5                   10                  15      


Leu Gln Thr Asp Ala Arg Asn Ser Asp Leu Thr Ile Thr Gln His Gly 
            20                  25                  30          


Gly Gly Asn Gly Ala Asp Val Gly Gln Gly Ser Asp Asp Ser Ser Ile 
        35                  40                  45              


Asp Leu Thr Gln Arg Gly Phe Gly Asn Ser Ala Thr Leu Asp Gln Trp 
    50                  55                  60                  


Asn Gly Lys Asn Ser Glu Met Thr Val Lys Gln Phe Gly Gly Gly Asn 
65                  70                  75                  80  


Gly Ala Ala Val Asp Gln Thr Ala Ser Asn 
                85                  90  


<210>  12
<211>  86
<212>  PRT
<213>  Escherichia coli

<400>  12

Ser Asp Leu Thr Ile Thr Gln His Gly Gly Gly Asn Gly Ala Asp Val 
1               5                   10                  15      


Gly Gln Gly Ser Asp Asp Ser Ser Ile Asp Leu Thr Gln Arg Gly Phe 
            20                  25                  30          


Gly Asn Ser Ala Thr Leu Asp Gln Trp Asn Gly Lys Asn Ser Glu Met 
        35                  40                  45              


Thr Val Lys Gln Phe Gly Gly Gly Asn Gly Ala Ala Val Asp Gln Thr 
    50                  55                  60                  


Ala Ser Asn Ser Ser Val Asn Val Thr Gln Val Gly Phe Gly Asn Asn 
65                  70                  75                  80  


Ala Thr Ala His Gln Tyr 
                85      


<210>  13
<211>  130
<212>  PRT
<213>  Lama glama

<400>  13

Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly 
1               5                   10                  15      


Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Gly Thr Asp Ser Asn Tyr 
            20                  25                  30          


Tyr Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Ile Val 
        35                  40                  45              


Ala Ala Ile Ser Trp Ile Gly Val Ile Glu Arg Tyr Thr Asp Ser Val 
    50                  55                  60                  


Lys Gly Arg Phe Thr Ile Ser Arg Glu Asn Ala Lys Asn Thr Val Ala 
65                  70                  75                  80  


Leu Gln Met Asn Ser Leu Asn Pro Glu Asp Thr Ala Val Tyr Tyr Cys 
                85                  90                  95      


Ala Ala Gly Arg Asn Asn Arg Gly Tyr Ser Asn Ser Trp Ser Arg Val 
            100                 105                 110         


Ala Ser Tyr Asp Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 
        115                 120                 125             


Gly Arg 
    130 


<210>  14
<211>  130
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  beta-lactamase

<400>  14

Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly 
1               5                   10                  15      


Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Gly Thr Asp Ser Asn Tyr 
            20                  25                  30          


Tyr Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Ile Val 
        35                  40                  45              


Ala Ala Ile Ser Trp Ile Gly Val Ile Glu Arg Tyr Thr Asp Ser Val 
    50                  55                  60                  


Lys Gly Arg Phe Thr Ile Ser Arg Glu Asn Ala Lys Asn Thr Val Ala 
65                  70                  75                  80  


Leu Gln Met Asn Ser Leu Asn Pro Glu Asp Thr Ala Val Tyr Tyr Cys 
                85                  90                  95      


Ala Ala Gly Arg Asn Asn Arg Gly Tyr Ser Asn Ser Trp Ser Arg Val 
            100                 105                 110         


Ala Ser Tyr Asp Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 
        115                 120                 125             


Gly Arg 
    130 


<210>  15
<211>  445
<212>  PRT
<213>  Escherichia coli

<400>  15

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


Gly Ala Arg Arg Leu Thr Gly Asp Gln Thr Ala Ala Leu Arg Asp Ser 
            20                  25                  30          


Leu Ser Asp Lys Pro Ala Lys Asn Ile Ile Leu Leu Ile Gly Asp Gly 
        35                  40                  45              


Met Gly Asp Ser Glu Ile Thr Ala Ala Arg Asn Tyr Ala Glu Gly Ala 
    50                  55                  60                  


Gly Gly Phe Phe Lys Gly Ile Asp Ala Leu Pro Leu Thr Gly Gln Tyr 
65                  70                  75                  80  


Thr His Tyr Ala Leu Asn Lys Lys Thr Gly Lys Pro Asp Tyr Val Thr 
                85                  90                  95      


Asp Ser Ala Ala Ser Ala Thr Ala Trp Ser Thr Gly Val Lys Thr Tyr 
            100                 105                 110         


Asn Gly Ala Leu Gly Val Asp Ile His Glu Lys Asp His Pro Thr Ile 
        115                 120                 125             


Leu Glu Met Ala Lys Ala Ala Gly Leu Ala Thr Gly Asn Val Ser Thr 
    130                 135                 140                 


Ala Glu Leu Gln Asp Ala Thr Pro Ala Ala Leu Val Ala His Val Thr 
145                 150                 155                 160 


Ser Arg Lys Cys Tyr Gly Pro Ser Ala Thr Ser Glu Lys Cys Pro Gly 
                165                 170                 175     


Asn Ala Leu Glu Lys Gly Gly Lys Gly Ser Ile Thr Glu Gln Leu Leu 
            180                 185                 190         


Asn Ala Arg Ala Asp Val Thr Leu Gly Gly Gly Ala Lys Thr Phe Ala 
        195                 200                 205             


Glu Thr Ala Thr Ala Gly Glu Trp Gln Gly Lys Thr Leu Arg Glu Gln 
    210                 215                 220                 


Ala Gln Ala Arg Gly Tyr Gln Leu Val Ser Asp Ala Ala Ser Leu Asn 
225                 230                 235                 240 


Ser Val Thr Glu Ala Asn Gln Gln Lys Pro Leu Leu Gly Leu Phe Ala 
                245                 250                 255     


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


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


Asn Asp Ser Val Pro Thr Leu Ala Gln Met Thr Asp Lys Ala Ile Glu 
    290                 295                 300                 


Leu Leu Ser Lys Asn Glu Lys Gly Phe Phe Leu Gln Val Glu Gly Ala 
305                 310                 315                 320 


Ser Ile Asp Lys Gln Asp His Ala Ala Asn Pro Cys Gly Gln Ile Gly 
                325                 330                 335     


Glu Thr Val Asp Leu Asp Glu Ala Val Gln Arg Ala Leu Glu Phe Ala 
            340                 345                 350         


Lys Lys Glu Gly Asn Thr Leu Val Ile Val Thr Ala Asp His Ala His 
        355                 360                 365             


Ala Ser Gln Ile Val Ala Pro Asp Thr Lys Ala Pro Gly Leu Thr Gln 
    370                 375                 380                 


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


Ser Glu Glu Asp Ser Gln Glu His Thr Gly Ser Gln Leu Arg Ile Ala 
                405                 410                 415     


Ala Tyr Gly Pro His Ala Ala Asn Val Val Gly Leu Thr Asp Gln Thr 
            420                 425                 430         


Asp Leu Phe Tyr Thr Met Lys Ala Ala Leu Gly Leu Lys 
        435                 440                 445 


<210>  16
<211>  205
<212>  PRT
<213>  Escherichia coli

<400>  16

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


Gln Glu Gln Leu Ala Val Thr Asn Asn Asp Glu Asn Ser Thr Tyr Leu 
            20                  25                  30          


Ile Gln Ser Trp Val Glu Asn Ala Asp Gly Val Lys Asp Gly Arg Phe 
        35                  40                  45              


Ile Val Thr Pro Pro Leu Phe Ala Met Lys Gly Lys Lys Glu Asn Thr 
    50                  55                  60                  


Leu Arg Ile Leu Asp Ala Thr Asn Asn Gln Leu Pro Gln Asp Arg Glu 
65                  70                  75                  80  


Ser Leu Phe Trp Met Asn Val Lys Ala Ile Pro Ser Met Asp Lys Ser 
                85                  90                  95      


Lys Leu Thr Glu Asn Thr Leu Gln Leu Ala Ile Ile Ser Arg Ile Lys 
            100                 105                 110         


Leu Tyr Tyr Arg Pro Ala Lys Leu Ala Leu Pro Pro Asp Gln Ala Ala 
        115                 120                 125             


Glu Lys Leu Arg Phe Arg Arg Ser Ala Asn Ser Leu Thr Leu Ile Asn 
    130                 135                 140                 


Pro Thr Pro Tyr Tyr Leu Thr Val Thr Glu Leu Asn Ala Gly Thr Arg 
145                 150                 155                 160 


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


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


Tyr Gly Ala Leu Thr Pro Lys Met Thr Gly Val Met Glu 
        195                 200                 205 


<210>  17
<211>  151
<212>  PRT
<213>  Escherichia coli

<400>  17

Asn Ser Ser Ala Ser Ser Ala Gln Val Thr Gly Thr Leu Leu Gly Thr 
1               5                   10                  15      


Gly Lys Thr Asn Thr Thr Gln Met Pro Ala Leu Tyr Thr Trp Gln His 
            20                  25                  30          


Gln Ile Tyr Asn Val Asn Phe Ile Pro Ser Ser Ser Gly Thr Leu Thr 
        35                  40                  45              


Cys Gln Ala Gly Thr Ile Leu Val Trp Lys Asn Gly Arg Glu Thr Gln 
    50                  55                  60                  


Tyr Ala Leu Glu Cys Arg Val Ser Ile His His Ser Ser Gly Ser Ile 
65                  70                  75                  80  


Asn Glu Ser Gln Trp Gly Gln Gln Ser Gln Val Gly Phe Gly Thr Ala 
                85                  90                  95      


Cys Gly Asn Lys Lys Cys Arg Phe Thr Gly Phe Glu Ile Ser Leu Arg 
            100                 105                 110         


Ile Pro Pro Asn Ala Gln Thr Tyr Pro Leu Ser Ser Gly Asp Leu Lys 
        115                 120                 125             


Gly Ser Phe Ser Leu Thr Asn Lys Glu Val Asn Trp Ser Ala Ser Ile 
    130                 135                 140                 


Tyr Val Pro Ala Ile Ala Lys 
145                 150     


<210>  18
<211>  245
<212>  PRT
<213>  Escherichia coli

<400>  18

Leu Ala Leu Gln Ala Lys Gln Tyr Gly Asp Phe Asp Arg Tyr Val Leu 
1               5                   10                  15      


Ala Leu Ser Trp Gln Thr Gly Phe Cys Gln Ser Gln His Asp Arg Asn 
            20                  25                  30          


Arg Asn Glu Arg Asp Glu Cys Arg Leu Gln Thr Glu Thr Thr Asn Lys 
        35                  40                  45              


Ala Asp Phe Leu Thr Val His Gly Leu Trp Pro Gly Leu Pro Lys Ser 
    50                  55                  60                  


Val Ala Ala Arg Gly Val Asp Glu Arg Arg Trp Met Arg Phe Gly Cys 
65                  70                  75                  80  


Ala Thr Arg Pro Ile Pro Asn Leu Pro Glu Ala Arg Ala Ser Arg Met 
                85                  90                  95      


Cys Ser Ser Pro Glu Thr Gly Leu Ser Leu Glu Thr Ala Ala Lys Leu 
            100                 105                 110         


Ser Glu Val Met Pro Gly Ala Gly Gly Arg Ser Cys Leu Glu Arg Tyr 
        115                 120                 125             


Glu Tyr Ala Lys His Gly Ala Cys Phe Gly Phe Asp Pro Asp Ala Tyr 
    130                 135                 140                 


Phe Gly Thr Met Val Arg Leu Asn Gln Glu Ile Lys Glu Ser Glu Ala 
145                 150                 155                 160 


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


Phe Asp Ala Ala Phe Ala Lys Ser Trp Gly Lys Glu Asn Val Lys Ala 
            180                 185                 190         


Val Lys Leu Thr Cys Gln Gly Asn Pro Ala Tyr Leu Thr Glu Ile Gln 
        195                 200                 205             


Ile Ser Ile Lys Ala Asp Ala Ile Asn Ala Pro Leu Ser Ala Asn Ser 
    210                 215                 220                 


Phe Leu Pro Gln Pro His Pro Gly Asn Cys Gly Lys Thr Phe Val Ile 
225                 230                 235                 240 


Asp Lys Ala Gly Tyr 
                245 


<210>  19
<211>  235
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  mCherry

<400>  19

Val Ser Lys Gly Glu Glu Asp Asn Met Ala Ile Ile Lys Glu Phe Met 
1               5                   10                  15      


Arg Phe Lys Val His Met Glu Gly Ser Val Asn Gly His Glu Phe Glu 
            20                  25                  30          


Ile Glu Gly Glu Gly Glu Gly Arg Pro Tyr Glu Gly Thr Gln Thr Ala 
        35                  40                  45              


Lys Leu Lys Val Thr Lys Gly Gly Pro Leu Pro Phe Ala Trp Asp Ile 
    50                  55                  60                  


Leu Ser Pro Gln Phe Met Tyr Gly Ser Lys Ala Tyr Val Lys His Pro 
65                  70                  75                  80  


Ala Asp Ile Pro Asp Tyr Leu Lys Leu Ser Phe Pro Glu Gly Phe Lys 
                85                  90                  95      


Trp Glu Arg Val Met Asn Phe Glu Asp Gly Gly Val Val Thr Val Thr 
            100                 105                 110         


Gln Asp Ser Ser Leu Gln Asp Gly Glu Phe Ile Tyr Lys Val Lys Leu 
        115                 120                 125             


Arg Gly Thr Asn Phe Pro Ser Asp Gly Pro Val Met Gln Lys Lys Thr 
    130                 135                 140                 


Met Gly Trp Glu Ala Ser Ser Glu Arg Met Tyr Pro Glu Asp Gly Ala 
145                 150                 155                 160 


Leu Lys Gly Glu Ile Lys Gln Arg Leu Lys Leu Lys Asp Gly Gly His 
                165                 170                 175     


Tyr Asp Ala Glu Val Lys Thr Thr Tyr Lys Ala Lys Lys Pro Val Gln 
            180                 185                 190         


Leu Pro Gly Ala Tyr Asn Val Asn Ile Lys Leu Asp Ile Thr Ser His 
        195                 200                 205             


Asn Glu Asp Tyr Thr Ile Val Glu Gln Tyr Glu Arg Ala Glu Gly Arg 
    210                 215                 220                 


His Ser Thr Gly Gly Met Asp Glu Leu Tyr Lys 
225                 230                 235 


<210>  20
<211>  260
<212>  PRT
<213>  Arabidopsis thaliana

<400>  20

Met Ala Glu Glu Tyr Lys Asn Thr Val Pro Glu Gln Glu Thr Pro Lys 
1               5                   10                  15      


Val Ala Thr Glu Glu Ser Ser Ala Pro Glu Ile Lys Glu Arg Gly Met 
            20                  25                  30          


Phe Asp Phe Leu Lys Lys Lys Glu Glu Val Lys Pro Gln Glu Thr Thr 
        35                  40                  45              


Thr Leu Ala Ser Glu Phe Glu His Lys Thr Gln Ile Ser Glu Pro Glu 
    50                  55                  60                  


Ser Phe Val Ala Lys His Glu Glu Glu Glu His Lys Pro Thr Leu Leu 
65                  70                  75                  80  


Glu Gln Leu His Gln Lys His Glu Glu Glu Glu Glu Asn Lys Pro Ser 
                85                  90                  95      


Leu Leu Asp Lys Leu His Arg Ser Asn Ser Ser Ser Ser Ser Ser Ser 
            100                 105                 110         


Asp Glu Glu Gly Glu Asp Gly Glu Lys Lys Lys Lys Glu Lys Lys Lys 
        115                 120                 125             


Lys Ile Val Glu Gly Asp His Val Lys Thr Val Glu Glu Glu Asn Gln 
    130                 135                 140                 


Gly Val Met Asp Arg Ile Lys Glu Lys Phe Pro Leu Gly Glu Lys Pro 
145                 150                 155                 160 


Gly Gly Asp Asp Val Pro Val Val Thr Thr Met Pro Ala Pro His Ser 
                165                 170                 175     


Val Glu Asp His Lys Pro Glu Glu Glu Glu Lys Lys Gly Phe Met Asp 
            180                 185                 190         


Lys Ile Lys Glu Lys Leu Pro Gly His Ser Lys Lys Pro Glu Asp Ser 
        195                 200                 205             


Gln Val Val Asn Thr Thr Pro Leu Val Glu Thr Ala Thr Pro Ile Ala 
    210                 215                 220                 


Asp Ile Pro Glu Glu Lys Lys Gly Phe Met Asp Lys Ile Lys Glu Lys 
225                 230                 235                 240 


Leu Pro Gly Tyr His Ala Lys Thr Thr Gly Glu Glu Glu Lys Lys Glu 
                245                 250                 255     


Lys Val Ser Asp 
            260 


<210>  21
<211>  28
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer

<400>  21
ccccggtacc cgttaatttc cattcgac                                          28


<210>  22
<211>  54
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer

<400>  22
cccctctaga ctaatggtga tggtgatggt gcccggggta ctgatgagcg atcg             54


<210>  23
<211>  48
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer

<400>  23
gctcatcagt acccctctgg ttctggttct ggtcaggtgc agctgcag                    48


<210>  24
<211>  151
<212>  PRT
<213>  Escherichia coli

<400>  24

Met Lys Asn Lys Leu Leu Phe Met Met Leu Thr Ile Leu Gly Ala Pro 
1               5                   10                  15      


Gly Ile Ala Ala Ala Ala Gly Tyr Asp Leu Ala Asn Ser Glu Tyr Asn 
            20                  25                  30          


Phe Ala Val Asn Glu Leu Ser Lys Ser Ser Phe Asn Gln Ala Ala Ile 
        35                  40                  45              


Ile Gly Gln Ala Gly Thr Asn Asn Ser Ala Gln Leu Arg Gln Gly Gly 
    50                  55                  60                  


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


Lys Ile Asp Gln Thr Gly Asp Tyr Asn Leu Ala Tyr Ile Asp Gln Ala 
                85                  90                  95      


Gly Ser Ala Asn Asp Ala Ser Ile Ser Gln Gly Ala Tyr Gly Asn Thr 
            100                 105                 110         


Ala Met Ile Ile Gln Lys Gly Ser Gly Asn Lys Ala Asn Ile Thr Gln 
        115                 120                 125             


Tyr Gly Thr Gln Lys Thr Ala Ile Val Val Gln Arg Gln Ser Gln Met 
    130                 135                 140                 


Ala Ile Arg Val Thr Gln Arg 
145                 150     


<210>  25
<211>  216
<212>  PRT
<213>  Escherichia coli

<400>  25

Met Phe Asn Glu Val His Ser Ile His Gly His Thr Leu Leu Leu Ile 
1               5                   10                  15      


Thr Lys Ser Ser Leu Gln Ala Thr Ala Leu Leu Gln His Leu Lys Gln 
            20                  25                  30          


Ser Leu Ala Ile Thr Gly Lys Leu His Asn Ile Gln Arg Ser Leu Asp 
        35                  40                  45              


Asp Ile Ser Ser Gly Ser Ile Ile Leu Leu Asp Met Met Glu Ala Asp 
    50                  55                  60                  


Lys Lys Leu Ile His Tyr Trp Gln Asp Thr Leu Ser Arg Lys Asn Asn 
65                  70                  75                  80  


Asn Ile Lys Ile Leu Leu Leu Asn Thr Pro Glu Asp Tyr Pro Tyr Arg 
                85                  90                  95      


Asp Ile Glu Asn Trp Pro His Ile Asn Gly Val Phe Tyr Ser Met Glu 
            100                 105                 110         


Asp Gln Glu Arg Val Val Asn Gly Leu Gln Gly Val Leu Arg Gly Glu 
        115                 120                 125             


Cys Tyr Phe Thr Gln Lys Leu Ala Ser Tyr Leu Ile Thr His Ser Gly 
    130                 135                 140                 


Asn Tyr Arg Tyr Asn Ser Thr Glu Ser Ala Leu Leu Thr His Arg Glu 
145                 150                 155                 160 


Lys Glu Ile Leu Asn Lys Leu Arg Ile Gly Ala Ser Asn Asn Glu Ile 
                165                 170                 175     


Ala Arg Ser Leu Phe Ile Ser Glu Asn Thr Val Lys Thr His Leu Tyr 
            180                 185                 190         


Asn Leu Phe Lys Lys Ile Ala Val Lys Asn Arg Thr Gln Ala Val Ser 
        195                 200                 205             


Trp Ala Asn Asp Asn Leu Arg Arg 
    210                 215     


<210>  26
<211>  129
<212>  PRT
<213>  Escherichia coli

<400>  26

Met Lys Arg Tyr Leu Arg Trp Ile Val Ala Ala Glu Phe Leu Phe Ala 
1               5                   10                  15      


Ala Gly Asn Leu His Ala Val Glu Val Glu Val Pro Gly Leu Leu Thr 
            20                  25                  30          


Asp His Thr Val Ser Ser Ile Gly His Asp Phe Tyr Arg Ala Phe Ser 
        35                  40                  45              


Asp Lys Trp Glu Ser Asp Tyr Thr Gly Asn Leu Thr Ile Asn Glu Arg 
    50                  55                  60                  


Pro Ser Ala Arg Trp Gly Ser Trp Ile Thr Ile Thr Val Asn Gln Asp 
65                  70                  75                  80  


Val Ile Phe Gln Thr Phe Leu Phe Pro Leu Lys Arg Asp Phe Glu Lys 
                85                  90                  95      


Thr Val Val Phe Ala Leu Ile Gln Thr Glu Glu Ala Leu Asn Arg Arg 
            100                 105                 110         


Gln Ile Asn Gln Ala Leu Leu Ser Thr Gly Asp Leu Ala His Asp Glu 
        115                 120                 125             


Phe 
    


<210>  27
<211>  138
<212>  PRT
<213>  Escherichia coli

<400>  27

Met Arg Val Lys His Ala Val Val Leu Leu Met Leu Ile Ser Pro Leu 
1               5                   10                  15      


Ser Trp Ala Gly Thr Met Thr Phe Gln Phe Arg Asn Pro Asn Phe Gly 
            20                  25                  30          


Gly Asn Pro Asn Asn Gly Ala Phe Leu Leu Asn Ser Ala Gln Ala Gln 
        35                  40                  45              


Asn Ser Tyr Lys Asp Pro Ser Tyr Asn Asp Asp Phe Gly Ile Glu Thr 
    50                  55                  60                  


Pro Ser Ala Leu Asp Asn Phe Thr Gln Ala Ile Gln Ser Gln Ile Leu 
65                  70                  75                  80  


Gly Gly Leu Leu Ser Asn Ile Asn Thr Gly Lys Pro Gly Arg Met Val 
                85                  90                  95      


Thr Asn Asp Tyr Ile Val Asp Ile Ala Asn Arg Asp Gly Gln Leu Gln 
            100                 105                 110         


Leu Asn Val Thr Asp Arg Lys Thr Gly Gln Thr Ser Thr Ile Gln Val 
        115                 120                 125             


Ser Gly Leu Gln Asn Asn Ser Thr Asp Phe 
    130                 135             


<210>  28
<211>  277
<212>  PRT
<213>  Escherichia coli

<400>  28

Met Gln Arg Leu Phe Leu Leu Val Ala Val Met Leu Leu Ser Gly Cys 
1               5                   10                  15      


Leu Thr Ala Pro Pro Lys Glu Ala Ala Arg Pro Thr Leu Met Pro Arg 
            20                  25                  30          


Ala Gln Ser Tyr Lys Asp Leu Thr His Leu Pro Ala Pro Thr Gly Lys 
        35                  40                  45              


Ile Phe Val Ser Val Tyr Asn Ile Gln Asp Glu Thr Gly Gln Phe Lys 
    50                  55                  60                  


Pro Tyr Pro Ala Ser Asn Phe Ser Thr Ala Val Pro Gln Ser Ala Thr 
65                  70                  75                  80  


Ala Met Leu Val Thr Ala Leu Lys Asp Ser Arg Trp Phe Ile Pro Leu 
                85                  90                  95      


Glu Arg Gln Gly Leu Gln Asn Leu Leu Asn Glu Arg Lys Ile Ile Arg 
            100                 105                 110         


Ala Ala Gln Glu Asn Gly Thr Val Ala Ile Asn Asn Arg Ile Pro Leu 
        115                 120                 125             


Gln Ser Leu Thr Ala Ala Asn Ile Met Val Glu Gly Ser Ile Ile Gly 
    130                 135                 140                 


Tyr Glu Ser Asn Val Lys Ser Gly Gly Val Gly Ala Arg Tyr Phe Gly 
145                 150                 155                 160 


Ile Gly Ala Asp Thr Gln Tyr Gln Leu Asp Gln Ile Ala Val Asn Leu 
                165                 170                 175     


Arg Val Val Asn Val Ser Thr Gly Glu Ile Leu Ser Ser Val Asn Thr 
            180                 185                 190         


Ser Lys Thr Ile Leu Ser Tyr Glu Val Gln Ala Gly Val Phe Arg Phe 
        195                 200                 205             


Ile Asp Tyr Gln Arg Leu Leu Glu Gly Glu Val Gly Tyr Thr Ser Asn 
    210                 215                 220                 


Glu Pro Val Met Leu Cys Leu Met Ser Ala Ile Glu Thr Gly Val Ile 
225                 230                 235                 240 


Phe Leu Ile Asn Asp Gly Ile Asp Arg Gly Leu Trp Asp Leu Gln Asn 
                245                 250                 255     


Lys Ala Glu Arg Gln Asn Asp Ile Leu Val Lys Tyr Arg His Met Ser 
            260                 265                 270         


Val Pro Pro Glu Ser 
        275         


<210>  29
<211>  18
<212>  PRT
<213>  Escherichia coli


<220>
<221>  misc_feature
<222>  (2)..(6)
<223>  Xaa can be any naturally occurring amino acid

<220>
<221>  misc_feature
<222>  (8)..(11)
<223>  Xaa can be any naturally occurring amino acid

<220>
<221>  misc_feature
<222>  (13)..(17)
<223>  Xaa can be any naturally occurring amino acid

<400>  29

Ser Xaa Xaa Xaa Xaa Xaa Gln Xaa Xaa Xaa Xaa Asn Xaa Xaa Xaa Xaa 
1               5                   10                  15      


Xaa Gln 
        


<210>  30
<211>  20
<212>  PRT
<213>  Escherichia coli

<400>  30

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


Ser Ala Leu Ala 
            20  


<210>  31
<211>  22
<212>  PRT
<213>  Escherichia coli

<400>  31

Gly Val Val Pro Gln Tyr Gly Gly Gly Gly Asn His Gly Gly Gly Gly 
1               5                   10                  15      


Asn Asn Ser Gly Pro Asn 
            20          


<210>  32
<211>  14
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Carrier polypeptide


<220>
<221>  MISC_FEATURE
<222>  (1)..(1)
<223>  X can be Val, Ile or Leu

<220>
<221>  misc_feature
<222>  (2)..(2)
<223>  Xaa can be any naturally occurring amino acid

<220>
<221>  misc_feature
<222>  (4)..(4)
<223>  Xaa can be any naturally occurring amino acid

<220>
<221>  misc_feature
<222>  (6)..(7)
<223>  Xaa can be any naturally occurring amino acid

<220>
<221>  MISC_FEATURE
<222>  (8)..(8)
<223>  X can be Asn or Gln

<220>
<221>  misc_feature
<222>  (9)..(9)
<223>  Xaa can be any naturally occurring amino acid

<220>
<221>  MISC_FEATURE
<222>  (10)..(10)
<223>  X can be Ala, Val, Ile or Leu

<220>
<221>  misc_feature
<222>  (11)..(13)
<223>  Xaa can be any naturally occurring amino acid

<400>  32

Xaa Xaa Gln Xaa Gly Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Gln 
1               5                   10                  


<210>  33
<211>  30
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  33
atggtgatgg tgcccgctgg agacggtgac                                        30


<210>  34
<211>  47
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  34
gctcatcagt acccctctgg ttctggttct ggtttagcgt tgcaggc                     47


<210>  35
<211>  30
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  35
atggtgatgg tgcccataac ccgctttatc                                        30


<210>  36
<211>  48
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  36
gctcatcagt acccctctgg ttctggttct ggtcctgttc tggaaaac                    48


<210>  37
<211>  30
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  37
atggtgatgg tgccctttca gccccagagc                                        30


<210>  38
<211>  50
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  38
gctcatcagt acccctctgg ttctggttct ggtaattcta gtgcgagtag                  50


<210>  39
<211>  30
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  39
atggtgatgg tgcccttttg caatcgcagg                                        30


<210>  40
<211>  49
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  40
gctcatcagt acccctctgg ttctggttct ggtcacccag aaacgctgg                   49


<210>  41
<211>  32
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  41
atggtgatgg tgcccccaat gcttaatcag tg                                     32


<210>  42
<211>  49
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  42
gctcatcagt acccctctgg ttctggttct ggtggagtgg ccttaggtg                   49


<210>  43
<211>  31
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  43
atggtgatgg tgcccttcca ttacgcccgt c                                      31


<210>  44
<211>  49
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  44
gctcatcagt acccctctgg ttctggttct ggtgtgagca agggcgagg                   49


<210>  45
<211>  31
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  45
atggtgatgg tgccccttgt acagctcgtc c                                      31


<210>  46
<211>  51
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  46
gctcatcagt acccctctgg ttctggttct ggtgcagaag agtacaagaa c                51


<210>  47
<211>  34
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  47
atggtgatgg tgcccatcag acactttttc tttc                                   34


<210>  48
<211>  34
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  48
ctctctgaga ctctctgctg tagcctctgg aggc                                   34


<210>  49
<211>  34
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  49
gcctccagag gctacagagg agagtctcag agag                                   34


<210>  50
<211>  28
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  50
gcagaagagt acaagaacac cgttccag                                          28


<210>  51
<211>  19
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  51
tgccagagcg ctaccggag                                                    19


<210>  52
<211>  67
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  52
aataactcaa ccgattttta agccccagct tcataaggaa aataatcgtg taggctggag       60

ctgcttc                                                                 67


<210>  53
<211>  67
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  53
cgcttaaaca gtaaaatgcc ggatgataat tccggctttt ttatctgcat atgaatatcc       60

tccttag                                                                 67


<210>  54
<211>  24
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  54
tctgagctga acatttacca gtac                                              24


<210>  55
<211>  19
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  55
tgccagagcg ctaccggag                                                    19


<210>  56
<211>  22
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  56
tctgacttga ctattaccca gc                                                22


<210>  57
<211>  23
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  57
atttgggccg ctattattac cgc                                               23


<210>  58
<211>  27
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  58
ccctctggtt ctggttctgg tcaggtg                                           27


<210>  59
<211>  21
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  59
gttagatgca gtctggtcaa c                                                 21


<210>  60
<211>  24
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  60
atttttgccg ttccactgat caag                                              24


<210>  61
<211>  18
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  61
gtcatctgag ccctgacc                                                     18


<210>  62
<211>  21
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  62
gttacgggca tcagtttgca g                                                 21


<210>  63
<211>  32
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  63
agctcaatcg atctgaccca acgtggcttc gg                                     32


<210>  64
<211>  29
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  64
tctgaaatga cggttaaaca gttcggtgg                                         29


<210>  65
<211>  27
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  65
tcctccgtca acgtgactca ggttggc                                           27


<210>  66
<211>  30
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  66
cccccatatg gttgttcctc agtacggcgg                                        30


<210>  67
<211>  52
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  67
ccccgaattc ctaatggtga tggtgaatgg tggtactgat gagcggtcgc gt               52


<210>  68
<211>  38
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  68
ggtgatggtg atggtgggat ccgtactgat gagcggtc                               38


<210>  69
<211>  38
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  69
gaccgctcat cagtacggat cccaccatca ccatcacc                               38


<210>  70
<211>  46
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  70
tcatcagtac ggatcctctg gttctggttc tggtcaggtg cagctg                      46


<210>  71
<211>  35
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  71
ggtgatggtg ggatccgctg gagacggtga cctgg                                  35


<210>  72
<211>  73
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  72
ggggacaagt ttgtacaaaa aagcaggctt gaaaggagga ataattaatg aaacttttaa       60

aagtagcagc aat                                                          73


<210>  73
<211>  52
<212>  DNA
<213>  artificial

<220>
<223>  Primer

<400>  73
ggggaccact ttgtacaaga aagctgggta ctaatggtga tggtgatggt gc               52


<210>  74
<211>  4
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Factor Xa cleavage site

<400>  74

Ile Glu Gly Arg 
1               


<210>  75
<211>  4
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  thrombin cleavage site

<400>  75

Leu Val Pro Arg 
1               


<210>  76
<211>  5
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  enterokinase cleaving site

<400>  76

Asp Asp Asp Asp Lys 
1               5   


<210>  77
<211>  8
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  PreScission cleavage site

<400>  77

Leu Glu Val Leu Phe Gln Gly Pro 
1               5               


