                         SEQUENCE LISTING

<110>  GeneOn Biotechnology
 
<120>  A MUTANT PET DEGRADING ENZYME

<130>  3

<160>  8     

<170>  PatentIn version 3.5

<210>  1
<211>  297
<212>  PRT
<213>  Ideonella sakaiensis

<400>  1

Met Met Asn Phe Pro Arg Ala Ser Arg Leu Met Gln Ala Ala Val Leu 
1               5                   10                  15      


Gly Gly Leu Met Ala Val Ser Ala Ala Ala Thr Ala Gln Thr Asn Pro 
            20                  25                  30          


Tyr Ala Arg Gly Pro Asn Pro Thr Ala Ala Ser Leu Glu Ala Ser Ala 
        35                  40                  45              


Gly Pro Phe Thr Val Arg Ser Phe Thr Val Ser Arg Pro Ser Gly Tyr 
    50                  55                  60                  


Gly Ala Gly Thr Val Tyr Tyr Pro Thr Asn Ala Gly Gly Thr Val Gly 
65                  70                  75                  80  


Ala Ile Ala Ile Val Pro Gly Tyr Thr Ala Arg Gln Ser Ser Ile Lys 
                85                  90                  95      


Trp Trp Gly Pro Arg Leu Ala Ser His Gly Phe Val Val Ile Thr Ile 
            100                 105                 110         


Asp Thr Asn Ser Thr Leu Asp Gln Pro Ser Ser Arg Ser Ser Gln Gln 
        115                 120                 125             


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


Pro Ile Tyr Gly Lys Val Asp Thr Ala Arg Met Gly Val Met Gly Trp 
145                 150                 155                 160 


Ser Met Gly Gly Gly Gly Ser Leu Ile Ser Ala Ala Asn Asn Pro Ser 
                165                 170                 175     


Leu Lys Ala Ala Ala Pro Gln Ala Pro Trp Asp Ser Ser Thr Asn Phe 
            180                 185                 190         


Ser Ser Val Thr Val Pro Thr Leu Ile Phe Ala Cys Glu Asn Asp Ser 
        195                 200                 205             


Ile Ala Pro Val Asn Ser Ser Ala Leu Pro Ile Tyr Asp Ser Met Ser 
    210                 215                 220                 


Arg Asn Ala Lys Gln Phe Leu Glu Ile Asn Gly Gly Ser His Ser Cys 
225                 230                 235                 240 


Ala Asn Ser Gly Asn Ser Asn Gln Ala Leu Ile Gly Lys Lys Gly Val 
                245                 250                 255     


Ala Trp Met Lys Arg Phe Met Asp Asn Asp Thr Arg Tyr Ser Thr Phe 
            260                 265                 270         


Ala Cys Glu Asn Pro Asn Ser Thr Arg Val Ser Asp Phe Arg Thr Ala 
        275                 280                 285             


Asn Cys Ser His His His His His His 
    290                 295         


<210>  2
<211>  297
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  mPETase Amino Acid Sequence

<400>  2

Met Met Asn Phe Pro Arg Ala Ser Arg Leu Met Gln Ala Ala Val Leu 
1               5                   10                  15      


Gly Gly Leu Met Ala Val Ser Ala Ala Ala Thr Ala Gln Thr Asn Pro 
            20                  25                  30          


Tyr Ala Arg Gly Pro Asn Pro Thr Ala Ala Ser Leu Glu Ala Ser Ala 
        35                  40                  45              


Gly Pro Phe Thr Val Arg Ser Phe Thr Val Ser Arg Pro Ser Gly Tyr 
    50                  55                  60                  


Gly Ala Gly Thr Val Tyr Tyr Pro Thr Asn Ala Gly Gly Thr Val Gly 
65                  70                  75                  80  


Ala Ile Ala Ile Val Pro Gly Tyr Thr Ala Arg Gln Ser Ser Ile Lys 
                85                  90                  95      


Trp Trp Gly Pro Arg Leu Ala Ser His Gly Phe Val Val Ile Thr Ile 
            100                 105                 110         


Asp Thr Asn Ser Thr Leu Asp Gln Pro Ser Ser Arg Ser Ser Gln Gln 
        115                 120                 125             


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


Pro Ile Tyr Gly Lys Val Asp Thr Ala Arg Met Gly Val Met Gly Trp 
145                 150                 155                 160 


Ser Met Gly Gly Gly Gly Ser Leu Ile Ser Ala Ala Asn Asn Pro Ser 
                165                 170                 175     


Leu Lys Ala Ala Ala Pro Gln Ala Pro Trp Asp Ser Ser Thr Asn Phe 
            180                 185                 190         


Ser Ser Val Thr Val Pro Thr Leu Ile Phe Ala Cys Glu Asn Asp Ser 
        195                 200                 205             


Ile Ala Pro Val Asn Ser Ser Ala Leu Pro Ile Tyr Asp Ser Met Ser 
    210                 215                 220                 


Arg Asn Ala Lys Gln Phe Leu Glu Ile Asn Gly Gly Ser His Ser Cys 
225                 230                 235                 240 


Ala Asn Ser Gly Asn Thr Asn Gln Ala Leu Ile Gly Lys Lys Gly Val 
                245                 250                 255     


Ala Trp Met Lys Arg Phe Met Asp Asn Asp Thr Arg Tyr Ser Thr Phe 
            260                 265                 270         


Ala Cys Glu Asn Pro Asn Ser Thr Arg Val Ser Asp Phe Arg Thr Ala 
        275                 280                 285             


Asn Cys Ser His His His His His His 
    290                 295         


<210>  3
<211>  915
<212>  DNA
<213>  Ideonella sakaiensis

<400>  3
taagcacata tgatgaattt cccccgtgcc tcgcgtttaa tgcaagctgc tgtcctcggc       60

ggtttaatgg ccgtatcagc tgccgccaca gcccaaacta atccttacgc acgtggtccc      120

aatcctacag cagcttccct ggaagcgtct gccggacctt tcacggttcg ttccttcacg      180

gtttcgcgtc cttcaggtta cggtgccggt acagtttact accccactaa tgccggtggt      240

acagttgggg caatcgctat cgttccagga tacacagccc gtcaatcgtc tatcaagtgg      300

tggggacctc gtttggccag tcacggtttc gtcgttatca cgatcgacac aaattcgacg      360

ctcgaccaac ccagttcacg tagttcccaa caaatggcag cattacgtca agttgcctcg      420

ttaaatggga cttcctcatc gcccatctac gggaaggttg acacggcacg tatgggtgta      480

atggggtggt ctatgggtgg tggtggatca cttatctccg ctgcaaataa tccaagtctt      540

aaggctgctg ccccgcaagc cccatgggac agttcaacga atttctcgtc agtcacggtt      600

ccaactttga tcttcgcttg tgagaatgac agtatcgccc ctgtaaattc atctgcactt      660

ccaatctacg acagtatgtc ccgtaatgcc aagcaattcc ttgagatcaa tggtgggtcg      720

cacagttgtg caaatagtgg aaatagcaat caagcattaa tcggtaagaa gggagttgcc      780

tggatgaagc gtttcatgga caatgacacg cgttactcga cattcgcctg tgagaatcca      840

aatagtactc gtgtaagtga cttccgtact gctaattgtt cgcatcacca ccatcatcac      900

tgaaagcttt aagca                                                       915


<210>  4
<211>  915
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  mPETase Nucleotide Sequence

<400>  4
taagcacata tgatgaattt cccccgtgcc tcgcgtttaa tgcaagctgc tgtcctcggc       60

ggtttaatgg ccgtatcagc tgccgccaca gcccaaacta atccttacgc acgtggtccc      120

aatcctacag cagcttccct ggaagcgtct gccggacctt tcacggttcg ttccttcacg      180

gtttcgcgtc cttcaggtta cggtgccggt acagtttact accccactaa tgccggtggt      240

acagttgggg caatcgctat cgttccagga tacacagccc gtcaatcgtc tatcaagtgg      300

tggggacctc gtttggccag tcacggtttc gtcgttatca cgatcgacac aaattcgacg      360

ctcgaccaac ccagttcacg tagttcccaa caaatggcag cattacgtca agttgcctcg      420

ttaaatggga cttcctcatc gcccatctac gggaaggttg acacggcacg tatgggtgta      480

atggggtggt ctatgggtgg tggtggatca cttatctccg ctgcaaataa tccaagtctt      540

aaggctgctg ccccgcaagc cccatgggac agttcaacga atttctcgtc agtcacggtt      600

ccaactttga tcttcgcttg tgagaatgac agtatcgccc ctgtaaattc atctgcactt      660

ccaatctacg acagtatgtc ccgtaatgcc aagcaattcc ttgagatcaa tggtgggtcg      720

cacagttgtg caaatagtgg aaataccaat caagcattaa tcggtaagaa gggagttgcc      780

tggatgaagc gtttcatgga caatgacacg cgttactcga cattcgcctg tgagaatcca      840

aatagtactc gtgtaagtga cttccgtact gctaattgtt cgcatcacca ccatcatcac      900

tgaaagcttt aagca                                                       915


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

<220>
<223>  T7 promoter forward primer

<400>  5
taatacgact cactataggg                                                   20


<210>  6
<211>  19
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  T7 terminator reverse primer

<400>  6
gctagttatt gctcagcgg                                                    19


<210>  7
<211>  50
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Site-directed mutagenesis Forward primer (S245T-F)

<400>  7
gggtcgcaca gttgtgcaaa tagtggaaat accaatcaag cattaatcgg                  50


<210>  8
<211>  50
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Site-directed mutagenesis Reverse primer (S245T-R)

<400>  8
ccgattaatg cttgattggt atttccacta tttgcacaac tgtgcgaccc                  50


