                         SEQUENCE LISTING

<110>  National Research Council of Canada
       Zwaagstraa, John C.
       O'Connor-McCourt, Maureen D.
       Sulea, Traian
       Collins, Catherine
       Banville, Myriam
       Jaramillo, Maria L.
 
<120>  COVALENTLY DIMERIZED BIVALENT BINDING AGENTS

<130>  12281-98

<150>  US 61/417,628
<151>  2010-11-29

<160>  26    

<170>  PatentIn version 3.5

<210>  1
<211>  7
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Heptad repeat used in E-coil

<400>  1

Glu Val Ser Ala Leu Glu Lys 
1               5           


<210>  2
<211>  7
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Heptad repeat used in E-coil

<400>  2

Glu Val Ser Ala Leu Glu Cys 
1               5           


<210>  3
<211>  7
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Heptad repeat used in E-coil

<400>  3

Glu Val Glu Ala Leu Glu Lys 
1               5           


<210>  4
<211>  7
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Heptad repeat used in E-coil

<400>  4

Glu Val Glu Ala Leu Gln Lys 
1               5           


<210>  5
<211>  7
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Heptad repeat used in K-coil

<400>  5

Lys Val Ser Ala Leu Lys Glu 
1               5           


<210>  6
<211>  7
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Heptad repeat used in K-coil

<400>  6

Lys Val Ser Ala Leu Lys Cys 
1               5           


<210>  7
<211>  7
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Heptad repeat used in K-coil

<400>  7

Lys Val Glu Ala Leu Lys Lys 
1               5           


<210>  8
<211>  6
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  TEV cleavage site

<400>  8

Glu Asn Leu Tyr Phe Gln 
1               5       


<210>  9
<211>  4
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Thrombin cleavage site

<400>  9

Phe Asn Pro Arg 
1               


<210>  10
<211>  4
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Factor Xa cleavage site


<220>
<221>  VARIANT
<222>  (2)..(2)
<223>  Xaa is glutamic acid or asparagine

<400>  10

Ile Xaa Gly Arg 
1               


<210>  11
<211>  5
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Enterokinase cleavage site

<400>  11

Asn Asn Asn Asn Lys 
1               5   


<210>  12
<211>  181
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  TbetaR2ECD with linker cassette

<400>  12

Met Gly Arg Gly Leu Leu Arg Gly Leu Trp Pro Leu His Ile Val Leu 
1               5                   10                  15      


Trp Thr Arg Ile Ala Ser Thr Ile Pro Pro Glu Gln Lys Leu Ile Ser 
            20                  25                  30          


Glu Glu Asp Leu Leu His Val Gln Lys Ser Val Asn Asn Asp Met Ile 
        35                  40                  45              


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


Cys Asp Val Arg Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser 
65                  70                  75                  80  


Asn Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val 
                85                  90                  95      


Ala Val Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys 
            100                 105                 110         


His Asp Pro Lys Leu Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala 
        115                 120                 125             


Ser Pro Lys Cys Ile Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe 
    130                 135                 140                 


Phe Met Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe 
145                 150                 155                 160 


Ser Glu Glu Tyr Asn Thr Ser Asn Pro Asp Gly Gly Arg Gly Gly Gly 
                165                 170                 175     


Gly Ser Gly Gly Gly 
            180     


<210>  13
<211>  56
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  cE5 fusion peptide

<400>  13

Cys Gly Gly Glu Asn Leu Tyr Phe Gln Gly Gly Gly Glu Val Ser Ala 
1               5                   10                  15      


Leu Glu Lys Glu Val Ser Ala Leu Glu Lys Glu Val Ser Ala Leu Glu 
            20                  25                  30          


Lys Glu Val Ser Ala Leu Glu Lys Glu Val Ser Ala Leu Glu Lys Gly 
        35                  40                  45              


Gly Gly His His His His His His 
    50                  55      


<210>  14
<211>  56
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  aE5 fusion peptide

<400>  14

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


Leu Glu Lys Glu Val Ser Ala Leu Glu Lys Glu Val Ser Ala Leu Glu 
            20                  25                  30          


Lys Glu Val Ser Ala Leu Glu Lys Glu Val Ser Ala Leu Glu Lys Gly 
        35                  40                  45              


Gly Gly His His His His His His 
    50                  55      


<210>  15
<211>  56
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  cK5 fusion peptide

<400>  15

Cys Gly Gly Glu Asn Leu Tyr Phe Gln Gly Gly Gly Lys Val Ser Ala 
1               5                   10                  15      


Leu Lys Glu Lys Val Ser Ala Leu Lys Glu Lys Val Ser Ala Leu Lys 
            20                  25                  30          


Glu Lys Val Ser Ala Leu Lys Glu Lys Val Ser Ala Leu Lys Glu Gly 
        35                  40                  45              


Gly Gly His His His His His His 
    50                  55      


<210>  16
<211>  56
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  aK5 fusion peptide

<400>  16

Ala Gly Gly Glu Asn Leu Tyr Phe Gln Gly Gly Gly Lys Val Ser Ala 
1               5                   10                  15      


Leu Lys Glu Lys Val Ser Ala Leu Lys Glu Lys Val Ser Ala Leu Lys 
            20                  25                  30          


Glu Lys Val Ser Ala Leu Lys Glu Lys Val Ser Ala Leu Lys Glu Gly 
        35                  40                  45              


Gly Gly His His His His His His 
    50                  55      


<210>  17
<211>  21
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  cdel fusion peptide

<400>  17

Cys Gly Gly Glu Asn Leu Tyr Phe Gln Gly Gly Gly Gly Gly Gly His 
1               5                   10                  15      


His His His His His 
            20      


<210>  18
<211>  21
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  adel fusion peptide

<400>  18

Ala Gly Gly Glu Asn Leu Tyr Phe Gln Gly Gly Gly Gly Gly Gly His 
1               5                   10                  15      


His His His His His 
            20      


<210>  19
<211>  220
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  R2Ec fusion peptide

<400>  19

Met Gly Arg Gly Leu Leu Arg Gly Leu Trp Pro Leu His Ile Val Leu 
1               5                   10                  15      


Trp Thr Arg Ile Ala Ser Thr Ile Pro Pro Glu Gln Lys Leu Ile Ser 
            20                  25                  30          


Glu Glu Asp Leu Leu His Val Gln Lys Ser Val Asn Asn Asp Met Ile 
        35                  40                  45              


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


Cys Asp Val Arg Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser 
65                  70                  75                  80  


Asn Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val 
                85                  90                  95      


Ala Val Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys 
            100                 105                 110         


His Asp Pro Lys Leu Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala 
        115                 120                 125             


Ser Pro Lys Cys Ile Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe 
    130                 135                 140                 


Phe Met Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe 
145                 150                 155                 160 


Ser Glu Glu Tyr Asn Thr Ser Asn Pro Asp Gly Gly Arg Gly Gly Gly 
                165                 170                 175     


Gly Ser Gly Gly Gly Glu Val Ser Ala Leu Glu Lys Glu Val Ser Ala 
            180                 185                 190         


Leu Glu Lys Glu Val Ser Ala Leu Glu Lys Glu Val Ser Ala Leu Glu 
        195                 200                 205             


Lys Glu Val Ser Ala Leu Glu Lys Gly Gly Gly Cys 
    210                 215                 220 


<210>  20
<211>  219
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  R2E fusion peptide

<400>  20

Met Gly Arg Gly Leu Leu Arg Gly Leu Trp Pro Leu His Ile Val Leu 
1               5                   10                  15      


Trp Thr Arg Ile Ala Ser Thr Ile Pro Pro Glu Gln Lys Leu Ile Ser 
            20                  25                  30          


Glu Glu Asp Leu Leu His Val Gln Lys Ser Val Asn Asn Asp Met Ile 
        35                  40                  45              


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


Cys Asp Val Arg Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser 
65                  70                  75                  80  


Asn Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val 
                85                  90                  95      


Ala Val Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys 
            100                 105                 110         


His Asp Pro Lys Leu Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala 
        115                 120                 125             


Ser Pro Lys Cys Ile Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe 
    130                 135                 140                 


Phe Met Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe 
145                 150                 155                 160 


Ser Glu Glu Tyr Asn Thr Ser Asn Pro Asp Gly Gly Arg Gly Gly Gly 
                165                 170                 175     


Gly Ser Gly Gly Gly Glu Val Ser Ala Leu Glu Lys Glu Val Ser Ala 
            180                 185                 190         


Leu Glu Lys Glu Val Ser Ala Leu Glu Lys Glu Val Ser Ala Leu Glu 
        195                 200                 205             


Lys Glu Val Ser Ala Leu Glu Lys Gly Gly Gly 
    210                 215                 


<210>  21
<211>  220
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  R2Kc fusion peptide

<400>  21

Met Gly Arg Gly Leu Leu Arg Gly Leu Trp Pro Leu His Ile Val Leu 
1               5                   10                  15      


Trp Thr Arg Ile Ala Ser Thr Ile Pro Pro Glu Gln Lys Leu Ile Ser 
            20                  25                  30          


Glu Glu Asp Leu Leu His Val Gln Lys Ser Val Asn Asn Asp Met Ile 
        35                  40                  45              


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


Cys Asp Val Arg Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser 
65                  70                  75                  80  


Asn Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val 
                85                  90                  95      


Ala Val Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys 
            100                 105                 110         


His Asp Pro Lys Leu Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala 
        115                 120                 125             


Ser Pro Lys Cys Ile Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe 
    130                 135                 140                 


Phe Met Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe 
145                 150                 155                 160 


Ser Glu Glu Tyr Asn Thr Ser Asn Pro Asp Gly Gly Arg Gly Gly Gly 
                165                 170                 175     


Gly Ser Gly Gly Gly Lys Val Ser Ala Leu Lys Glu Lys Val Ser Ala 
            180                 185                 190         


Leu Lys Glu Lys Val Ser Ala Leu Lys Glu Lys Val Ser Ala Leu Lys 
        195                 200                 205             


Glu Lys Val Ser Ala Leu Lys Glu Gly Gly Gly Cys 
    210                 215                 220 


<210>  22
<211>  219
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  R2K fusion peptide

<400>  22

Met Gly Arg Gly Leu Leu Arg Gly Leu Trp Pro Leu His Ile Val Leu 
1               5                   10                  15      


Trp Thr Arg Ile Ala Ser Thr Ile Pro Pro Glu Gln Lys Leu Ile Ser 
            20                  25                  30          


Glu Glu Asp Leu Leu His Val Gln Lys Ser Val Asn Asn Asp Met Ile 
        35                  40                  45              


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


Cys Asp Val Arg Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser 
65                  70                  75                  80  


Asn Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val 
                85                  90                  95      


Ala Val Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys 
            100                 105                 110         


His Asp Pro Lys Leu Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala 
        115                 120                 125             


Ser Pro Lys Cys Ile Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe 
    130                 135                 140                 


Phe Met Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe 
145                 150                 155                 160 


Ser Glu Glu Tyr Asn Thr Ser Asn Pro Asp Gly Gly Arg Gly Gly Gly 
                165                 170                 175     


Gly Ser Gly Gly Gly Lys Val Ser Ala Leu Lys Glu Lys Val Ser Ala 
            180                 185                 190         


Leu Lys Glu Lys Val Ser Ala Leu Lys Glu Lys Val Ser Ala Leu Lys 
        195                 200                 205             


Glu Lys Val Ser Ala Leu Lys Glu Gly Gly Gly 
    210                 215                 


<210>  23
<211>  182
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  R1Ec fusion peptide

<400>  23

Met Glu Ala Ala Ser Ala Ala Leu Arg Arg Cys Leu Leu Leu Ile Val 
1               5                   10                  15      


Leu Val Ala Ala Ala Thr Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu 
            20                  25                  30          


Leu Pro Gly Ala Lys Ala Leu Gln Cys Phe Cys His Leu Cys Thr Lys 
        35                  40                  45              


Asp Asn Phe Thr Cys Glu Thr Asp Gly Leu Cys Phe Val Ser Val Thr 
    50                  55                  60                  


Glu Thr Thr Asp Lys Val Ile His Asn Ser Met Cys Ile Ala Glu Ile 
65                  70                  75                  80  


Asp Leu Ile Pro Arg Asp Arg Pro Phe Val Cys Ala Pro Ser Ser Lys 
                85                  90                  95      


Thr Gly Ala Val Thr Tyr Cys Cys Asn Gln Asp His Cys Asn Lys Ile 
            100                 105                 110         


Glu Leu Pro Thr Thr Gly Pro Phe Ser Glu Lys Gln Ser Ala Gly Leu 
        115                 120                 125             


Gly Pro Val Glu Gly Gly Arg Gly Gly Gly Gly Ser Gly Gly Gly Glu 
    130                 135                 140                 


Val Ser Ala Leu Glu Lys Glu Val Ser Ala Leu Glu Lys Glu Val Ser 
145                 150                 155                 160 


Ala Leu Glu Lys Glu Val Ser Ala Leu Glu Lys Glu Val Ser Ala Leu 
                165                 170                 175     


Glu Lys Gly Gly Gly Cys 
            180         


<210>  24
<211>  181
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  R1E fusion peptide

<400>  24

Met Glu Ala Ala Ser Ala Ala Leu Arg Arg Cys Leu Leu Leu Ile Val 
1               5                   10                  15      


Leu Val Ala Ala Ala Thr Glu Gln Lys Leu Ile Ser Glu Glu Asp Leu 
            20                  25                  30          


Leu Pro Gly Ala Lys Ala Leu Gln Cys Phe Cys His Leu Cys Thr Lys 
        35                  40                  45              


Asp Asn Phe Thr Cys Glu Thr Asp Gly Leu Cys Phe Val Ser Val Thr 
    50                  55                  60                  


Glu Thr Thr Asp Lys Val Ile His Asn Ser Met Cys Ile Ala Glu Ile 
65                  70                  75                  80  


Asp Leu Ile Pro Arg Asp Arg Pro Phe Val Cys Ala Pro Ser Ser Lys 
                85                  90                  95      


Thr Gly Ala Val Thr Tyr Cys Cys Asn Gln Asp His Cys Asn Lys Ile 
            100                 105                 110         


Glu Leu Pro Thr Thr Gly Pro Phe Ser Glu Lys Gln Ser Ala Gly Leu 
        115                 120                 125             


Gly Pro Val Glu Gly Gly Arg Gly Gly Gly Gly Ser Gly Gly Gly Glu 
    130                 135                 140                 


Val Ser Ala Leu Glu Lys Glu Val Ser Ala Leu Glu Lys Glu Val Ser 
145                 150                 155                 160 


Ala Leu Glu Lys Glu Val Ser Ala Leu Glu Lys Glu Val Ser Ala Leu 
                165                 170                 175     


Glu Lys Gly Gly Gly 
            180     


<210>  25
<211>  235
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  R2cK

<400>  25

Met Gly Arg Gly Leu Leu Arg Gly Leu Trp Pro Leu His Ile Val Leu 
1               5                   10                  15      


Trp Thr Arg Ile Ala Ser Thr Ile Pro Pro Glu Gln Lys Leu Ile Ser 
            20                  25                  30          


Glu Glu Asp Leu Leu His Val Gln Lys Ser Val Asn Asn Asp Met Ile 
        35                  40                  45              


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


Cys Asp Val Arg Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser 
65                  70                  75                  80  


Asn Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val 
                85                  90                  95      


Ala Val Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys 
            100                 105                 110         


His Asp Pro Lys Leu Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala 
        115                 120                 125             


Ser Pro Lys Cys Ile Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe 
    130                 135                 140                 


Phe Met Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe 
145                 150                 155                 160 


Ser Glu Glu Tyr Asn Thr Ser Asn Pro Asp Gly Gly Arg Gly Gly Gly 
                165                 170                 175     


Gly Ser Gly Gly Gly Cys Gly Gly Phe Asn Pro Arg Gly Gly Gly Lys 
            180                 185                 190         


Val Ser Ala Leu Lys Glu Lys Val Ser Ala Leu Lys Glu Lys Val Ser 
        195                 200                 205             


Ala Leu Lys Glu Lys Val Ser Ala Leu Lys Glu Lys Val Ser Ala Leu 
    210                 215                 220                 


Lys Glu Gly Gly Gly His His His His His His 
225                 230                 235 


<210>  26
<211>  235
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  R2cE

<400>  26

Met Gly Arg Gly Leu Leu Arg Gly Leu Trp Pro Leu His Ile Val Leu 
1               5                   10                  15      


Trp Thr Arg Ile Ala Ser Thr Ile Pro Pro Glu Gln Lys Leu Ile Ser 
            20                  25                  30          


Glu Glu Asp Leu Leu His Val Gln Lys Ser Val Asn Asn Asp Met Ile 
        35                  40                  45              


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


Cys Asp Val Arg Phe Ser Thr Cys Asp Asn Gln Lys Ser Cys Met Ser 
65                  70                  75                  80  


Asn Cys Ser Ile Thr Ser Ile Cys Glu Lys Pro Gln Glu Val Cys Val 
                85                  90                  95      


Ala Val Trp Arg Lys Asn Asp Glu Asn Ile Thr Leu Glu Thr Val Cys 
            100                 105                 110         


His Asp Pro Lys Leu Pro Tyr His Asp Phe Ile Leu Glu Asp Ala Ala 
        115                 120                 125             


Ser Pro Lys Cys Ile Met Lys Glu Lys Lys Lys Pro Gly Glu Thr Phe 
    130                 135                 140                 


Phe Met Cys Ser Cys Ser Ser Asp Glu Cys Asn Asp Asn Ile Ile Phe 
145                 150                 155                 160 


Ser Glu Glu Tyr Asn Thr Ser Asn Pro Asp Gly Gly Arg Gly Gly Gly 
                165                 170                 175     


Gly Ser Gly Gly Gly Cys Gly Gly Phe Asn Pro Arg Gly Gly Gly Glu 
            180                 185                 190         


Val Ser Ala Leu Glu Lys Glu Val Ser Ala Leu Glu Lys Glu Val Ser 
        195                 200                 205             


Ala Leu Glu Lys Glu Val Ser Ala Leu Glu Lys Glu Val Ser Ala Leu 
    210                 215                 220                 


Glu Lys Gly Gly Gly His His His His His His 
225                 230                 235 


