                         SEQUENCE LISTING

<110>  The University of Manchester
 
<120>  METHODS FOR ENZYME ENGINEERING

<130>  P328769WO

<150>  2108011.4
<151>  2021-06-04

<160>  7     

<170>  PatentIn version 3.5

<210>  1
<211>  1380
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  HDNO D2

<400>  1
atggttagca gcaaactggc aacaccgctg agcattcagg gtgaagttat ttatccggat       60

gatagcggtt ttgatgccat tgccaatatt tgggatggtc gtcatctgca gcgtccgagc      120

ctgattgcac gttgtctgag tgccggtgat gttgcaaaaa gcgttcgtta tgcatgtgat      180

aatggtctgg aaatttcagt tcgtagcggt ggtcataatc cgaatggtta tgcaaccaat      240

gatggtggta ttgttctgga tctgcgtctg atgaatagca ttcatattga taccgcaggt      300

agccgtgcac gtattggtgg tggtgttatt agcggtgatc tggttaaaga agcagcaaaa      360

tttggtctgg cagcagttac cggtatgcat ccgaaagttg gtttttgtgg tctggccctg      420

aatggtggtg tgggttttct gaccccgaaa tatggcctgg caagcgataa cattctgggt      480

gcaaccctgg ttaccgcaac aggtgatgtg atttattgta gtgatgatga acgtccggaa      540

ctgttttggg cagttcgtgg tgcaggtccg aattttggtg ttgttaccga agttgaagtt      600

cagctgtatg aactgcctcg taaaatgctg gcaggtttta ttacctgggc accgagcgtt      660

agcgaactgg caggtctgct gaccagcctg ctggatgcac tgaatgaaat ggcagatcat      720

atctatccga gcgtttttgt tggtgtggat gaaaatcgtg caccgagtgt taccgtttgt      780

gttggtcatc tgggtggtct ggatattgca gaacgtgata ttgcacgtct gcgtggcctg      840

ggtcgtaccg ttagcgatag cattgccgtt cgtagctatg atgaagttgt tgcgctgaat      900

gcagaagttg gtagctttga agatggtatg agcaatctgt ggattgatcg tgaaattgca      960

atgccgaatg cacgttttgc agaagcaatt gcaggtaacc tggataaatt tgtgagcgaa     1020

ccggcaagcg gtggtagcgt taaactgttg attgatggta tgccgtttgg taatccgaaa     1080

cgtacaccgg cacgtcatcg tgatgcaatg ggtgttctgg cactggcaga atggtcaggt     1140

gcagcaccgg gtagcgagaa atatcctgaa ctggcacgtg aactggatgc agcactgctg     1200

cgtgcgggtg ttaccaccag tggttttggc ctgctgaata ataacagcga agttaccgca     1260

gaaatggttg ccgaagtgta taaaccggaa gtttatagtc gcctggcagc cgttaaacgt     1320

gaatatgatc cggaaaatcg ttttcgccac aactataaca tcgatccgga aggtagctaa     1380


<210>  2
<211>  459
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  HDNO D2

<400>  2

Met Val Ser Ser Lys Leu Ala Thr Pro Leu Ser Ile Gln Gly Glu Val 
1               5                   10                  15      


Ile Tyr Pro Asp Asp Ser Gly Phe Asp Ala Ile Ala Asn Ile Trp Asp 
            20                  25                  30          


Gly Arg His Leu Gln Arg Pro Ser Leu Ile Ala Arg Cys Leu Ser Ala 
        35                  40                  45              


Gly Asp Val Ala Lys Ser Val Arg Tyr Ala Cys Asp Asn Gly Leu Glu 
    50                  55                  60                  


Ile Ser Val Arg Ser Gly Gly His Asn Pro Asn Gly Tyr Ala Thr Asn 
65                  70                  75                  80  


Asp Gly Gly Ile Val Leu Asp Leu Arg Leu Met Asn Ser Ile His Ile 
                85                  90                  95      


Asp Thr Ala Gly Ser Arg Ala Arg Ile Gly Gly Gly Val Ile Ser Gly 
            100                 105                 110         


Asp Leu Val Lys Glu Ala Ala Lys Phe Gly Leu Ala Ala Val Thr Gly 
        115                 120                 125             


Met His Pro Lys Val Gly Phe Cys Gly Leu Ala Leu Asn Gly Gly Val 
    130                 135                 140                 


Gly Phe Leu Thr Pro Lys Tyr Gly Leu Ala Ser Asp Asn Ile Leu Gly 
145                 150                 155                 160 


Ala Thr Leu Val Thr Ala Thr Gly Asp Val Ile Tyr Cys Ser Asp Asp 
                165                 170                 175     


Glu Arg Pro Glu Leu Phe Trp Ala Val Arg Gly Ala Gly Pro Asn Phe 
            180                 185                 190         


Gly Val Val Thr Glu Val Glu Val Gln Leu Tyr Glu Leu Pro Arg Lys 
        195                 200                 205             


Met Leu Ala Gly Phe Ile Thr Trp Ala Pro Ser Val Ser Glu Leu Ala 
    210                 215                 220                 


Gly Leu Leu Thr Ser Leu Leu Asp Ala Leu Asn Glu Met Ala Asp His 
225                 230                 235                 240 


Ile Tyr Pro Ser Val Phe Val Gly Val Asp Glu Asn Arg Ala Pro Ser 
                245                 250                 255     


Val Thr Val Cys Val Gly His Leu Gly Gly Leu Asp Ile Ala Glu Arg 
            260                 265                 270         


Asp Ile Ala Arg Leu Arg Gly Leu Gly Arg Thr Val Ser Asp Ser Ile 
        275                 280                 285             


Ala Val Arg Ser Tyr Asp Glu Val Val Ala Leu Asn Ala Glu Val Gly 
    290                 295                 300                 


Ser Phe Glu Asp Gly Met Ser Asn Leu Trp Ile Asp Arg Glu Ile Ala 
305                 310                 315                 320 


Met Pro Asn Ala Arg Phe Ala Glu Ala Ile Ala Gly Asn Leu Asp Lys 
                325                 330                 335     


Phe Val Ser Glu Pro Ala Ser Gly Gly Ser Val Lys Leu Leu Ile Asp 
            340                 345                 350         


Gly Met Pro Phe Gly Asn Pro Lys Arg Thr Pro Ala Arg His Arg Asp 
        355                 360                 365             


Ala Met Gly Val Leu Ala Leu Ala Glu Trp Ser Gly Ala Ala Pro Gly 
    370                 375                 380                 


Ser Glu Lys Tyr Pro Glu Leu Ala Arg Glu Leu Asp Ala Ala Leu Leu 
385                 390                 395                 400 


Arg Ala Gly Val Thr Thr Ser Gly Phe Gly Leu Leu Asn Asn Asn Ser 
                405                 410                 415     


Glu Val Thr Ala Glu Met Val Ala Glu Val Tyr Lys Pro Glu Val Tyr 
            420                 425                 430         


Ser Arg Leu Ala Ala Val Lys Arg Glu Tyr Asp Pro Glu Asn Arg Phe 
        435                 440                 445             


Arg His Asn Tyr Asn Ile Asp Pro Glu Gly Ser 
    450                 455                 


<210>  3
<211>  1380
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  HDNO D3

<400>  3
atggttagca gcaaactggc aacaccgctg agcattcagg gtgaagttat ttatccggat       60

gatagcggtt ttgatgccat tgccaatatt tgggatggtc gtcatctgca gcgtccgagc      120

ctgattgcac gttgtctgag tgccggtgat gttgcaaaaa gcgttcgtta tgcatgtgat      180

aatggtctgg aaatttcagt tcgtagcggt ggtcataatc cgaatggtta tgcaaccaat      240

gatggtggta ttgttctgga tctgcgtctg atgaatagca ttcatattga taccgcaggt      300

agccgtgcac gtattggtgg tggtgttatt agcggtgatc tggttaaaga agcagcaaaa      360

tttggtctgg cagcagttac cggtatgcat ccgaaagttg gtttttgtgg tctggccctg      420

aatggtggtg tgggttttct gaccccgaaa tatggcctgg caagcgataa cattctgggt      480

gcaaccctgg ttaccgcaac aggtgatgtg atttattgta gtgatgatga acgtccggaa      540

ctgttttggg cagttcgtgg tgcaggtccg aattttggtg ttgttaccga agttgaagtt      600

cagctgtatg aactgcctcg taaaatgctg gcaggtttta ttacctgggc accgagcgtt      660

agcgaactgg caggtctgct gaccagcctg ctggatgcac tgaatgaaat ggcagatcat      720

atctatccga gcgtttttgt tggtgtggat gaaaatcgtg caccgagtgt taccgtttgt      780

gttggtcatc tgggtggtct ggatattgca gaacgtgata ttgcacgtct gcgtggcctg      840

ggtcgtaccg ttagcgatag cattgccgtt cgtagctatg atgaagttgt tgcgctgaat      900

gcagaagttg gtagctttga agatggtatg agcaatctgt ggattgatcg tgaaattgca      960

atgccgaatg cacgttttgc agaagcaatt gcaggtaacc tggataaatt tgtgagcgaa     1020

ccggcaagcg gtggtagcgt taaactgttg attgatggta tgccgtttgg taatccgaaa     1080

cgtacaccgg cacgtcatcg tgatgcaatg ggtgttctgg cactggcaga atggtcaggt     1140

gcagcaccgg gtagcgagaa atatcctgaa ctggcacgtg aactggatgc agcactgctg     1200

cgtgcgggtg ttaccaccag tggttttggc ctgctgaatc ataacagcga agttaccgca     1260

gaaatggttg ccgaagtgta taaaccggaa gtttatagtc gcctggcagc cgttaaacgt     1320

gaatatgatc cggaaaatcg ttttcgccac aactataaca tcgatccgga aggtagctaa     1380


<210>  4
<211>  459
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  HDNO D3

<400>  4

Met Val Ser Ser Lys Leu Ala Thr Pro Leu Ser Ile Gln Gly Glu Val 
1               5                   10                  15      


Ile Tyr Pro Asp Asp Ser Gly Phe Asp Ala Ile Ala Asn Ile Trp Asp 
            20                  25                  30          


Gly Arg His Leu Gln Arg Pro Ser Leu Ile Ala Arg Cys Leu Ser Ala 
        35                  40                  45              


Gly Asp Val Ala Lys Ser Val Arg Tyr Ala Cys Asp Asn Gly Leu Glu 
    50                  55                  60                  


Ile Ser Val Arg Ser Gly Gly His Asn Pro Asn Gly Tyr Ala Thr Asn 
65                  70                  75                  80  


Asp Gly Gly Ile Val Leu Asp Leu Arg Leu Met Asn Ser Ile His Ile 
                85                  90                  95      


Asp Thr Ala Gly Ser Arg Ala Arg Ile Gly Gly Gly Val Ile Ser Gly 
            100                 105                 110         


Asp Leu Val Lys Glu Ala Ala Lys Phe Gly Leu Ala Ala Val Thr Gly 
        115                 120                 125             


Met His Pro Lys Val Gly Phe Cys Gly Leu Ala Leu Asn Gly Gly Val 
    130                 135                 140                 


Gly Phe Leu Thr Pro Lys Tyr Gly Leu Ala Ser Asp Asn Ile Leu Gly 
145                 150                 155                 160 


Ala Thr Leu Val Thr Ala Thr Gly Asp Val Ile Tyr Cys Ser Asp Asp 
                165                 170                 175     


Glu Arg Pro Glu Leu Phe Trp Ala Val Arg Gly Ala Gly Pro Asn Phe 
            180                 185                 190         


Gly Val Val Thr Glu Val Glu Val Gln Leu Tyr Glu Leu Pro Arg Lys 
        195                 200                 205             


Met Leu Ala Gly Phe Ile Thr Trp Ala Pro Ser Val Ser Glu Leu Ala 
    210                 215                 220                 


Gly Leu Leu Thr Ser Leu Leu Asp Ala Leu Asn Glu Met Ala Asp His 
225                 230                 235                 240 


Ile Tyr Pro Ser Val Phe Val Gly Val Asp Glu Asn Arg Ala Pro Ser 
                245                 250                 255     


Val Thr Val Cys Val Gly His Leu Gly Gly Leu Asp Ile Ala Glu Arg 
            260                 265                 270         


Asp Ile Ala Arg Leu Arg Gly Leu Gly Arg Thr Val Ser Asp Ser Ile 
        275                 280                 285             


Ala Val Arg Ser Tyr Asp Glu Val Val Ala Leu Asn Ala Glu Val Gly 
    290                 295                 300                 


Ser Phe Glu Asp Gly Met Ser Asn Leu Trp Ile Asp Arg Glu Ile Ala 
305                 310                 315                 320 


Met Pro Asn Ala Arg Phe Ala Glu Ala Ile Ala Gly Asn Leu Asp Lys 
                325                 330                 335     


Phe Val Ser Glu Pro Ala Ser Gly Gly Ser Val Lys Leu Leu Ile Asp 
            340                 345                 350         


Gly Met Pro Phe Gly Asn Pro Lys Arg Thr Pro Ala Arg His Arg Asp 
        355                 360                 365             


Ala Met Gly Val Leu Ala Leu Ala Glu Trp Ser Gly Ala Ala Pro Gly 
    370                 375                 380                 


Ser Glu Lys Tyr Pro Glu Leu Ala Arg Glu Leu Asp Ala Ala Leu Leu 
385                 390                 395                 400 


Arg Ala Gly Val Thr Thr Ser Gly Phe Gly Leu Leu Asn His Asn Ser 
                405                 410                 415     


Glu Val Thr Ala Glu Met Val Ala Glu Val Tyr Lys Pro Glu Val Tyr 
            420                 425                 430         


Ser Arg Leu Ala Ala Val Lys Arg Glu Tyr Asp Pro Glu Asn Arg Phe 
        435                 440                 445             


Arg His Asn Tyr Asn Ile Asp Pro Glu Gly Ser 
    450                 455                 


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

<220>
<223>  look up table sequence

<400>  5

Gly Met Phe Trp Lys Ala Ile Cys 
1               5               


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

<220>
<223>  First 8 residues removed from PDB sequence

<400>  6

Met Glu Leu Gln Gln Val Leu Arg 
1               5               


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

<220>
<223>  start of PDB protein sequence

<400>  7

Met Asn Gly Gly 
1               


