                         SEQUENCE LISTING

<110>  Oxford University Innovation Limited
 
<120>  Polypeptide with enhanced rate of spontaneous isopeptide bond 
       formation with its peptide tag partner and uses thereof

<130>  20.11.142409/01

<150>  GB 1903479.2
<151>  2019-03-14

<160>  13    

<170>  PatentIn version 3.5

<210>  1
<211>  113
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  SpyCatcher003

<400>  1

Val Thr Thr Leu Ser Gly Leu Ser Gly Glu Gln Gly Pro Ser Gly Asp 
1               5                   10                  15      


Met Thr Thr Glu Glu Asp Ser Ala Thr His Ile Lys Phe Ser Lys Arg 
            20                  25                  30          


Asp Glu Asp Gly Arg Glu Leu Ala Gly Ala Thr Met Glu Leu Arg Asp 
        35                  40                  45              


Ser Ser Gly Lys Thr Ile Ser Thr Trp Ile Ser Asp Gly His Val Lys 
    50                  55                  60                  


Asp Phe Tyr Leu Tyr Pro Gly Lys Tyr Thr Phe Val Glu Thr Ala Ala 
65                  70                  75                  80  


Pro Asp Gly Tyr Glu Val Ala Thr Pro Ile Glu Phe Thr Val Asn Glu 
                85                  90                  95      


Asp Gly Gln Val Thr Val Asp Gly Glu Ala Thr Glu Gly Asp Ala His 
            100                 105                 110         


Thr 
    


<210>  2
<211>  92
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Truncated SpyCatcher003

<400>  2

Asp Ser Ala Thr His Ile Lys Phe Ser Lys Arg Asp Glu Asp Gly Arg 
1               5                   10                  15      


Glu Leu Ala Gly Ala Thr Met Glu Leu Arg Asp Ser Ser Gly Lys Thr 
            20                  25                  30          


Ile Ser Thr Trp Ile Ser Asp Gly His Val Lys Asp Phe Tyr Leu Tyr 
        35                  40                  45              


Pro Gly Lys Tyr Thr Phe Val Glu Thr Ala Ala Pro Asp Gly Tyr Glu 
    50                  55                  60                  


Val Ala Thr Pro Ile Glu Phe Thr Val Asn Glu Asp Gly Gln Val Thr 
65                  70                  75                  80  


Val Asp Gly Glu Ala Thr Glu Gly Asp Ala His Thr 
                85                  90          


<210>  3
<211>  16
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  SpyTag003

<400>  3

Arg Gly Val Pro His Ile Val Met Val Asp Ala Tyr Lys Arg Tyr Lys 
1               5                   10                  15      


<210>  4
<211>  14
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  SpyTag002

<400>  4

Val Pro Thr Ile Val Met Val Asp Ala Tyr Lys Arg Tyr Lys 
1               5                   10                  


<210>  5
<211>  13
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  SpyTag

<400>  5

Ala His Ile Val Met Val Asp Ala Tyr Lys Pro Thr Lys 
1               5                   10              


<210>  6
<211>  16
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  SpyTag variant


<220>
<221>  MISC_FEATURE
<222>  (1)..(1)
<223>  X is arginine or no amino acid

<220>
<221>  MISC_FEATURE
<222>  (1)..(2)
<223>  When X at position 1 is no amino acid, X at position 2 is no 
       amino acid

<220>
<221>  MISC_FEATURE
<222>  (2)..(2)
<223>  X is glycine or no amino acid

<220>
<221>  MISC_FEATURE
<222>  (5)..(5)
<223>  X is histidine or threonine, preferably hisitidine

<220>
<221>  MISC_FEATURE
<222>  (11)..(11)
<223>  X is alanine, glycine or valine, preferably alanine

<220>
<221>  MISC_FEATURE
<222>  (14)..(14)
<223>  X is arginine or lysine, preferably arginine

<400>  6

Xaa Xaa Val Pro Xaa Ile Val Met Val Asp Xaa Tyr Lys Xaa Tyr Lys 
1               5                   10                  15      


<210>  7
<211>  113
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  SpyCatcher002

<400>  7

Val Thr Thr Leu Ser Gly Leu Ser Gly Glu Gln Gly Pro Ser Gly Asp 
1               5                   10                  15      


Met Thr Thr Glu Glu Asp Ser Ala Thr His Ile Lys Phe Ser Lys Arg 
            20                  25                  30          


Asp Glu Asp Gly Arg Glu Leu Ala Gly Ala Thr Met Glu Leu Arg Asp 
        35                  40                  45              


Ser Ser Gly Lys Thr Ile Ser Thr Trp Ile Ser Asp Gly His Val Lys 
    50                  55                  60                  


Asp Phe Tyr Leu Tyr Pro Gly Lys Tyr Thr Phe Val Glu Thr Ala Ala 
65                  70                  75                  80  


Pro Asp Gly Tyr Glu Val Ala Thr Ala Ile Thr Phe Thr Val Asn Glu 
                85                  90                  95      


Gln Gly Gln Val Thr Val Asn Gly Glu Ala Thr Lys Gly Asp Ala His 
            100                 105                 110         


Thr 
    


<210>  8
<211>  113
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  SpyCatcher

<400>  8

Val Asp Thr Leu Ser Gly Leu Ser Ser Glu Gln Gly Gln Ser Gly Asp 
1               5                   10                  15      


Met Thr Ile Glu Glu Asp Ser Ala Thr His Ile Lys Phe Ser Lys Arg 
            20                  25                  30          


Asp Glu Asp Gly Lys Glu Leu Ala Gly Ala Thr Met Glu Leu Arg Asp 
        35                  40                  45              


Ser Ser Gly Lys Thr Ile Ser Thr Trp Ile Ser Asp Gly Gln Val Lys 
    50                  55                  60                  


Asp Phe Tyr Leu Tyr Pro Gly Lys Tyr Thr Phe Val Glu Thr Ala Ala 
65                  70                  75                  80  


Pro Asp Gly Tyr Glu Val Ala Thr Ala Ile Thr Phe Thr Val Asn Glu 
                85                  90                  95      


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


Ile 
    


<210>  9
<211>  339
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Bacterial codon optimised SpyCatcher003

<400>  9
gtaaccacct tatcaggttt atcaggtgag caaggtccgt ccggtgatat gacaactgaa       60

gaagatagtg ctacccatat taaattctca aaacgtgatg aggacggccg tgagttagct      120

ggtgcaacta tggagttgcg tgattcatct ggtaaaacta ttagtacatg gatttcagat      180

ggacatgtga aggatttcta cctgtatcca ggaaaatata catttgtcga aaccgcagca      240

ccagacggtt atgaggtagc aactccaatt gaatttacag ttaatgagga cggtcaggtt      300

actgtagatg gtgaagcaac tgaaggtgac gctcatact                             339


<210>  10
<211>  339
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Mammallian codon optimised SpyCatcher003

<400>  10
gtgaccacac tgtccggact gtctggagag cagggaccat ccggcgacat gaccacagag       60

gaggattctg ccacacacat caagttcagc aagagggacg aggacggaag agagctggca      120

ggagcaacca tggagctgag ggatagctcc ggcaagacca tcagcacatg gatctccgac      180

ggccacgtga aggatttcta cctgtatccc ggcaagtaca cctttgtgga gacagcagca      240

ccagacggat atgaggtggc aacccctatc gagtttacag tgaacgagga cggacaggtg      300

accgtggatg gagaggcaac agagggcgat gcacacacc                             339


<210>  11
<211>  16
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  SpyTag003-DA

<400>  11

Arg Gly Val Pro His Ile Val Met Val Ala Ala Tyr Lys Arg Tyr Lys 
1               5                   10                  15      


<210>  12
<211>  113
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  SpyCatcher003 S49C

<400>  12

Val Thr Thr Leu Ser Gly Leu Ser Gly Glu Gln Gly Pro Ser Gly Asp 
1               5                   10                  15      


Met Thr Thr Glu Glu Asp Ser Ala Thr His Ile Lys Phe Ser Lys Arg 
            20                  25                  30          


Asp Glu Asp Gly Arg Glu Leu Ala Gly Ala Thr Met Glu Leu Arg Asp 
        35                  40                  45              


Cys Ser Gly Lys Thr Ile Ser Thr Trp Ile Ser Asp Gly His Val Lys 
    50                  55                  60                  


Asp Phe Tyr Leu Tyr Pro Gly Lys Tyr Thr Phe Val Glu Thr Ala Ala 
65                  70                  75                  80  


Pro Asp Gly Tyr Glu Val Ala Thr Pro Ile Glu Phe Thr Val Asn Glu 
                85                  90                  95      


Asp Gly Gln Val Thr Val Asp Gly Glu Ala Thr Glu Gly Asp Ala His 
            100                 105                 110         


Thr 
    


<210>  13
<211>  92
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Truncated SpyCatcher003 S28C

<400>  13

Asp Ser Ala Thr His Ile Lys Phe Ser Lys Arg Asp Glu Asp Gly Arg 
1               5                   10                  15      


Glu Leu Ala Gly Ala Thr Met Glu Leu Arg Asp Cys Ser Gly Lys Thr 
            20                  25                  30          


Ile Ser Thr Trp Ile Ser Asp Gly His Val Lys Asp Phe Tyr Leu Tyr 
        35                  40                  45              


Pro Gly Lys Tyr Thr Phe Val Glu Thr Ala Ala Pro Asp Gly Tyr Glu 
    50                  55                  60                  


Val Ala Thr Pro Ile Glu Phe Thr Val Asn Glu Asp Gly Gln Val Thr 
65                  70                  75                  80  


Val Asp Gly Glu Ala Thr Glu Gly Asp Ala His Thr 
                85                  90          


