                         SEQUENCE LISTING

<110>  EXTEND BIOPHARMA, INC.
 
<120>  METHODS FOR SELECTING EVOLVED TRANSPEPTIDASES AND USE THEREOF FOR
        CREATING PROTEIN CONJUGATES

<130>  95489-945392

<140>  PCT/US2015/032717
<141>  2015-05-27

<150>  US 62/003,495
<151>  2014-05-27

<160>  34    

<170>  PatentIn version 3.5

<210>  1
<211>  206
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic wt S. aureus Sortase A

<400>  1

Met Lys Lys Trp Thr Asn Arg Leu Met Thr Ile Ala Gly Val Val Leu 
1               5                   10                  15      


Ile Leu Val Ala Ala Tyr Leu Phe Ala Lys Pro His Ile Asp Asn Tyr 
            20                  25                  30          


Leu His Asp Lys Asp Lys Asp Glu Lys Ile Glu Gln Tyr Asp Lys Asn 
        35                  40                  45              


Val Lys Glu Gln Ala Ser Lys Asp Lys Lys Gln Gln Ala Lys Pro Gln 
    50                  55                  60                  


Ile Pro Lys Asp Lys Ser Lys Val Ala Gly Tyr Ile Glu Ile Pro Asp 
65                  70                  75                  80  


Ala Asp Ile Lys Glu Pro Val Tyr Pro Gly Pro Ala Thr Pro Glu Gln 
                85                  90                  95      


Leu Asn Arg Gly Val Ser Phe Ala Glu Glu Asn Glu Ser Leu Asp Asp 
            100                 105                 110         


Gln Asn Ile Ser Ile Ala Gly His Thr Phe Ile Asp Arg Pro Asn Tyr 
        115                 120                 125             


Gln Phe Thr Asn Leu Lys Ala Ala Lys Lys Gly Ser Met Val Tyr Phe 
    130                 135                 140                 


Lys Val Gly Asn Glu Thr Arg Lys Tyr Lys Met Thr Ser Ile Arg Asp 
145                 150                 155                 160 


Val Lys Pro Thr Asp Val Glu Val Leu Asp Glu Gln Lys Gly Lys Asp 
                165                 170                 175     


Lys Gln Leu Thr Leu Ile Thr Cys Asp Asp Tyr Asn Glu Lys Thr Gly 
            180                 185                 190         


Val Trp Glu Lys Arg Lys Ile Phe Val Ala Thr Glu Val Lys 
        195                 200                 205     


<210>  2
<211>  5
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic Sortase A  recognition  sequence


<220>
<221>  VARIANT
<222>  (3)..(3)
<223>  X = any amino acid

<400>  2

Leu Pro Xaa Thr Gly 
1               5   


<210>  3
<211>  5
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic Sortase A  recognition  sequence

<400>  3

Leu Ser Pro Gly Lys 
1               5   


<210>  4
<211>  5
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic acceptor sequence motifs


<220>
<221>  VARIANT
<222>  (4)..(4)
<223>  X = any amino acid

<400>  4

Asp Ile Gln Xaa Thr 
1               5   


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

<220>
<223>  Synthetic acceptor sequence motifs

<400>  5

Asp Ile Gln Met Thr 
1               5   


<210>  6
<211>  5
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic acceptor sequence motifs

<400>  6

Asp Ile Gln Leu Thr 
1               5   


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

<220>
<223>  Synthetic acceptor sequence motifs


<220>
<221>  VARIANT
<222>  (1)..(1)
<223>  X = any amino acid

<220>
<221>  VARIANT
<222>  (5)..(5)
<223>  X = any amino acid

<400>  7

Xaa Val Gln Leu Xaa 
1               5   


<210>  8
<211>  5
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic acceptor sequence motifs

<400>  8

Glu Val Gln Leu Val 
1               5   


<210>  9
<211>  5
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic acceptor sequence motifs

<400>  9

Gln Val Gln Leu Val 
1               5   


<210>  10
<211>  5
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic acceptor sequence motifs

<400>  10

Gln Val Gln Leu Gln 
1               5   


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

<220>
<223>  Synthetic fluorescence resonance energy transfer (FRET) 
       substrates

<400>  11

Leu Pro Glu Thr Gly 
1               5   


<210>  12
<211>  7
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic TPase mutant primer

<400>  12

Cys Leu Pro Glu Thr Gly Gly 
1               5           


<210>  13
<211>  6
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Sytnthetic nucleophile

<400>  13

Gly Gly Gly Gly Ser Lys 
1               5       


<210>  14
<211>  5
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic Sortase B recognition sequence

<400>  14

Asn Pro Gln Thr Asn 
1               5   


<210>  15
<211>  5
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic target recognition  sequence

<400>  15

Gly Gly Gly Gly Gly 
1               5   


<210>  16
<211>  5
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic Sortase A  recognition  sequence


<220>
<221>  VARIANT
<222>  (3)..(3)
<223>  X = any amino acid

<400>  16

Leu Pro Xaa Thr Ala 
1               5   


<210>  17
<211>  7
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic Sortase A  recognition  sequence


<220>
<221>  MISC_FEATURE
<222>  (7)..(7)
<223>  X = Biotin

<400>  17

Gly Gly Gly Gly Gly Lys Xaa 
1               5           


<210>  18
<211>  5
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic acceptor sequence motifs

<400>  18

Asp Ile Leu Leu Thr 
1               5   


<210>  19
<211>  5
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic acceptor sequence motifs

<400>  19

Gln Ile Val Ser Thr 
1               5   


<210>  20
<211>  5
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic acceptor sequence motifs

<400>  20

Gln Ile Val Leu Ser 
1               5   


<210>  21
<211>  5
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic acceptor sequence motifs

<400>  21

Gln Ile Val Leu Thr 
1               5   


<210>  22
<211>  5
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic acceptor sequence motifs

<400>  22

Asp Val Val Met Thr 
1               5   


<210>  23
<211>  5
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic acceptor sequence motifs

<400>  23

Asp Val Gln Leu Thr 
1               5   


<210>  24
<211>  110
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic Linker


<220>
<221>  VARIANT
<222>  (6)..(110)
<223>  amino acids may be deleted by 5 sequential  residue increments at
       the C-terminus  up to 21 increments

<400>  24

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 
1               5                   10                  15      


Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 
            20                  25                  30          


Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 
        35                  40                  45              


Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 
    50                  55                  60                  


Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 
65                  70                  75                  80  


Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 
                85                  90                  95      


Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 
            100                 105                 110 


<210>  25
<211>  112
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic Linker


<220>
<221>  VARIANT
<222>  (5)..(112)
<223>  amino acids may be deleted by 4 sequential  residue increments at
       the C-terminus  up to 27 increments

<400>  25

Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser 
1               5                   10                  15      


Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser 
            20                  25                  30          


Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser 
        35                  40                  45              


Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser 
    50                  55                  60                  


Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser 
65                  70                  75                  80  


Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser 
                85                  90                  95      


Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser Gly Gly Gly Ser 
            100                 105                 110         


<210>  26
<211>  111
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic Linker


<220>
<221>  VARIANT
<222>  (4)..(11)
<223>  amino acids may be deleted by 3 sequential  residue increments at
       the C-terminus  up to 36 increments

<400>  26

Gly Gly Ser Gly Gly Ser Gly Gly Ser Gly Gly Ser Gly Gly Ser Gly 
1               5                   10                  15      


Gly Ser Gly Gly Ser Gly Gly Ser Gly Gly Ser Gly Gly Ser Gly Gly 
            20                  25                  30          


Ser Gly Gly Ser Gly Gly Ser Gly Gly Ser Gly Gly Ser Gly Gly Ser 
        35                  40                  45              


Gly Gly Ser Gly Gly Ser Gly Gly Ser Gly Gly Ser Gly Gly Ser Gly 
    50                  55                  60                  


Gly Ser Gly Gly Ser Gly Gly Ser Gly Gly Ser Gly Gly Ser Gly Gly 
65                  70                  75                  80  


Ser Gly Gly Ser Gly Gly Ser Gly Gly Ser Gly Gly Ser Gly Gly Ser 
                85                  90                  95      


Gly Gly Ser Gly Gly Ser Gly Gly Ser Gly Gly Ser Gly Gly Ser 
            100                 105                 110     


<210>  27
<211>  110
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic Linker


<220>
<221>  VARIANT
<222>  (3)..(110)
<223>  amino acids may be deleted by 2 sequential  residue increments at
       the C-terminus  up to 54 increments

<400>  27

Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser 
1               5                   10                  15      


Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser 
            20                  25                  30          


Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser 
        35                  40                  45              


Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser 
    50                  55                  60                  


Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser 
65                  70                  75                  80  


Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser 
                85                  90                  95      


Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser Gly Ser 
            100                 105                 110 


<210>  28
<211>  110
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic Linker


<220>
<221>  VARIANT
<222>  (1)..(110)
<223>  amino acids 1-110 represent a repeated variant that may be 
       repeated from 1 to 55 times as long as the total length is not 
       more than 110 residue

<220>
<221>  VARIANT
<222>  (2)..(109)
<223>  X = G or absent

<400>  28

Gly Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 
1               5                   10                  15      


Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 
            20                  25                  30          


Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 
        35                  40                  45              


Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 
    50                  55                  60                  


Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 
65                  70                  75                  80  


Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 
                85                  90                  95      


Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Ser 
            100                 105                 110 


<210>  29
<211>  110
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic Linker


<220>
<221>  VARIANT
<222>  (1)..(109)
<223>  X = any amino acid

<220>
<221>  VARIANT
<222>  (1)..(110)
<223>  amino acids 1-110 represent a repeated variant that may be 
       repeated from 1 to 55 times as long as the total length is not 
       more than 110 residue

<400>  29

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 
1               5                   10                  15      


Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 
            20                  25                  30          


Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 
        35                  40                  45              


Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 
    50                  55                  60                  


Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 
65                  70                  75                  80  


Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 
                85                  90                  95      


Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Ser 
            100                 105                 110 


<210>  30
<211>  5
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic Linker

<400>  30

Gly Gly Gly Gly Ser 
1               5   


<210>  31
<211>  6
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic small peptide recognition site of wt sortase

<400>  31

Cys Leu Pro Glu Thr Gly 
1               5       


<210>  32
<211>  6
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic S. Aureus Sortase A recognition site

<400>  32

Leu Pro Glu Thr Gly Gly 
1               5       


<210>  33
<211>  7
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic S. Aureus Sortase A recognition site

<400>  33

Leu Ser Leu Ser Pro Gly Lys 
1               5           


<210>  34
<211>  4
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic Tpase

<400>  34

Gly Gly Gly Gly 
1               


