                         SEQUENCE LISTING

<110>  Katholieke Universiteit Leuven K.U.Leuven R&D
       University of Minho
 
<120>  NOVEL ENDOLYSIN

<130>  LYS-014 PCT

<140>  not yet known
<141>  2011-11-03

<150>  GB 1018518.9
<151>  2010-11-03

<160>  31    

<170>  PatentIn version 3.3

<210>  1
<211>  235
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  wt PVPSE1gp146

<400>  1

Met Asn Ala Ala Ile Ala Glu Ile Gln Arg Met Leu Ile Glu Gly Gly 
1               5                   10                  15      


Phe Ser Val Gly Lys Ser Gly Ala Asp Gly Leu Tyr Gly Pro Ala Thr 
            20                  25                  30          


Lys Ala Ala Leu Gln Lys Cys Ile Ala Gln Ala Thr Ser Gly Asn Asn 
        35                  40                  45              


Lys Gly Gly Thr Leu Lys Leu Thr Gln Ala Gln Leu Asp Lys Ile Phe 
    50                  55                  60                  


Pro Val Gly Ala Ser Ser Gly Arg Asn Ala Lys Leu Lys Pro Leu Asn 
65                  70                  75                  80  


Asp Leu Phe Glu Lys Thr Glu Ile Asn Thr Val Asn Arg Val Ala Gly 
                85                  90                  95      


Phe Leu Ser Gln Ile Gly Val Glu Ser Ala Glu Phe Arg Tyr Val Arg 
            100                 105                 110         


Glu Leu Gly Asn Asp Ala Tyr Phe Asp Lys Tyr Asp Thr Gly Pro Ile 
        115                 120                 125             


Ala Glu Arg Leu Gly Asn Thr Pro Gln Lys Asp Gly Asp Gly Ala Lys 
    130                 135                 140                 


Tyr Lys Gly Arg Gly Leu Ile Gln Val Thr Gly Leu Ala Asn Tyr Lys 
145                 150                 155                 160 


Ala Cys Gly Lys Ala Leu Gly Leu Asp Leu Val Asn His Pro Glu Leu 
                165                 170                 175     


Leu Glu Gln Pro Glu Tyr Ala Val Ala Ser Ala Gly Trp Tyr Trp Asp 
            180                 185                 190         


Thr Arg Asn Ile Asn Ala Ala Cys Asp Ala Asp Asp Ile Val Lys Ile 
        195                 200                 205             


Thr Lys Leu Val Asn Gly Gly Thr Asn His Leu Ala Glu Arg Thr Ala 
    210                 215                 220                 


Tyr Tyr Lys Lys Ala Lys Ser Val Leu Thr Ser 
225                 230                 235 


<210>  2
<211>  711
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  wt ORF146 of PVPSE1

<400>  2
atgaatgctg caattgcgga gattcagcgt atgctgatcg aaggtgggtt tagcgtcggc     60

aagtctggtg ctgatggatt gtacggaccc gctacaaaag ccgcactgca aaagtgcatt    120

gcacaggcta ccagtggaaa caataaagga ggtactttga aactcaccca agcacaactg    180

gacaaaatct tccccgttgg tgcaagttct gggaggaatg caaaattcct gaagccgctc    240

aatgacctgt ttgaaaagac agagattaat acggtaaatc gggttgcagg attcctgtct    300

cagattggtg tggagtcggc ggagttccgg tatgtacgtg aactcggtaa cgatgcctac    360

tttgacaagt acgacactgg tcctattgca gaaagacttg gaaacacacc ccagaaagat    420

ggggatggtg ccaagtacaa ggggagaggt ctgattcagg tgaccggact cgcaaactac    480

aaggcttgcg gtaaagcact cggtcttgac ctcgttaacc accctgagtt gcttgaacag    540

cctgagtatg cagttgccag cgctggttgg tattgggaca cgagaaacat caacgccgct    600

tgcgatgctg atgatatcgt gaaaattacc aagctggtaa acggtggtac aaatcacctt    660

gccgagcgca cagcctatta caaaaaggca aaatctgttc taacctcgta a             711


<210>  3
<211>  37
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  peptidoglycan binding domain of PVPSE1

<400>  3

Ala Ala Ile Ala Glu Ile Gln Arg Met Leu Ile Glu Gly Gly Phe Ser 
1               5                   10                  15      


Val Gly Lys Ser Gly Ala Asp Gly Leu Tyr Gly Pro Ala Thr Lys Ala 
            20                  25                  30          


Ala Leu Gln Lys Cys 
        35          


<210>  4
<211>  111
<212>  DNA
<213>  Artificial sequence

<220>
<223>  pepdidoglycan binding domain of PVPSE1

<400>  4
gctgcaattg cggagattca gcgtatgctg atcgaaggtg ggtttagcgt cggcaagtct     60

ggtgctgatg gattgtacgg acccgctaca aaagccgcac tgcaaaagtg c             111


<210>  5
<211>  153
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  catalytic domain of PVPSE1

<400>  5

Asp Leu Phe Glu Lys Thr Glu Ile Asn Thr Val Asn Arg Val Ala Gly 
1               5                   10                  15      


Phe Leu Ser Gln Ile Gly Val Glu Ser Ala Glu Phe Arg Tyr Val Arg 
            20                  25                  30          


Glu Leu Gly Asn Asp Ala Tyr Phe Asp Lys Tyr Asp Thr Gly Pro Ile 
        35                  40                  45              


Ala Glu Arg Leu Gly Asn Thr Pro Gln Lys Asp Gly Asp Gly Ala Lys 
    50                  55                  60                  


Tyr Lys Gly Arg Gly Leu Ile Gln Val Thr Gly Leu Ala Asn Tyr Lys 
65                  70                  75                  80  


Ala Cys Gly Lys Ala Leu Gly Leu Asp Leu Val Asn His Pro Glu Leu 
                85                  90                  95      


Leu Glu Gln Pro Glu Tyr Ala Val Ala Ser Ala Gly Trp Tyr Trp Asp 
            100                 105                 110         


Thr Arg Asn Ile Asn Ala Ala Cys Asp Ala Asp Asp Ile Val Lys Ile 
        115                 120                 125             


Thr Lys Leu Val Asn Gly Gly Thr Asn His Leu Ala Glu Arg Thr Ala 
    130                 135                 140                 


Tyr Tyr Lys Lys Ala Lys Ser Val Leu 
145                 150             


<210>  6
<211>  459
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  catalytic domain of PVPSE1

<400>  6
gacctgtttg aaaagacaga gattaatacg gtaaatcggg ttgcaggatt cctgtctcag     60

attggtgtgg agtcggcgga gttccggtat gtacgtgaac tcggtaacga tgcctacttt    120

gacaagtacg acactggtcc tattgcagaa agacttggaa acacacccca gaaagatggg    180

gatggtgcca agtacaaggg gagaggtctg attcaggtga ccggactcgc aaactacaag    240

gcttgcggta aagcactcgg tcttgacctc gttaaccacc ctgagttgct tgaacagcct    300

gagtatgcag ttgccagcgc tggttggtat tgggacacga gaaacatcaa cgccgcttgc    360

gatgctgatg atatcgtgaa aattaccaag ctggtaaacg gtggtacaaa tcaccttgcc    420

gagcgcacag cctattacaa aaaggcaaaa tctgttcta                           459


<210>  7
<211>  245
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  recombinant PVPSE1gp146

<400>  7

Met Gly Ser Asn Ala Ala Ile Ala Glu Ile Gln Arg Met Leu Ile Glu 
1               5                   10                  15      


Gly Gly Phe Ser Val Gly Lys Ser Gly Ala Asp Gly Leu Tyr Gly Pro 
            20                  25                  30          


Ala Thr Lys Ala Ala Leu Gln Lys Cys Ile Ala Gln Ala Thr Ser Gly 
        35                  40                  45              


Asn Asn Lys Gly Gly Thr Leu Lys Leu Thr Gln Ala Gln Leu Asp Lys 
    50                  55                  60                  


Ile Phe Pro Val Gly Ala Ser Ser Gly Arg Asn Ala Lys Leu Lys Pro 
65                  70                  75                  80  


Leu Asn Asp Leu Phe Glu Lys Thr Glu Ile Asn Thr Val Asn Arg Val 
                85                  90                  95      


Ala Gly Phe Leu Ser Gln Ile Gly Val Glu Ser Ala Glu Phe Arg Tyr 
            100                 105                 110         


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


Pro Ile Ala Glu Arg Leu Gly Asn Thr Pro Gln Lys Asp Gly Asp Gly 
    130                 135                 140                 


Ala Lys Tyr Lys Gly Arg Gly Leu Ile Gln Val Thr Gly Leu Ala Asn 
145                 150                 155                 160 


Tyr Lys Ala Cys Gly Lys Ala Leu Gly Leu Asp Leu Val Asn His Pro 
                165                 170                 175     


Glu Leu Leu Glu Gln Pro Glu Tyr Ala Val Ala Ser Ala Gly Trp Tyr 
            180                 185                 190         


Trp Asp Thr Arg Asn Ile Asn Ala Ala Cys Asp Ala Asp Asp Ile Val 
        195                 200                 205             


Lys Ile Thr Lys Leu Val Asn Gly Gly Thr Asn His Leu Ala Glu Arg 
    210                 215                 220                 


Thr Ala Tyr Tyr Lys Lys Ala Lys Ser Val Leu Thr Ser Lys Gly His 
225                 230                 235                 240 


His His His His His 
                245 


<210>  8
<211>  741
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  recombinant PVPSE1gp146

<400>  8
atgggatcca atgctgcaat tgcggagatt cagcgtatgc tgatcgaagg tgggtttagc     60

gtcggcaagt ctggtgctga tggattgtac ggacccgcta caaaagccgc actgcaaaag    120

tgcattgcac aggctaccag tggaaacaat aaaggaggta ctttgaaact cacccaagca    180

caactggaca aaatcttccc cgttggtgca agttctggga ggaatgcaaa attcctgaag    240

ccgctcaatg acctgtttga aaagacagag attaatacgg taaatcgggt tgcaggattc    300

ctgtctcaga ttggtgtgga gtcggcggag ttccggtatg tacgtgaact cggtaacgat    360

gcctactttg acaagtacga cactggtcct attgcagaaa gacttggaaa cacaccccag    420

aaagatgggg atggtgccaa gtacaagggg agaggtctga ttcaggtgac cggactcgca    480

aactacaagg cttgcggtaa agcactcggt cttgacctcg ttaaccaccc tgagttgctt    540

gaacagcctg agtatgcagt tgccagcgct ggttggtatt gggacacgag aaacatcaac    600

gccgcttgcg atgctgatga tatcgtgaaa attaccaagc tggtaaacgg tggtacaaat    660

caccttgccg agcgcacagc ctattacaaa aaggcaaaat ctgttctaac ctcgaagggt    720

catcatcacc atcaccattg a                                              741


<210>  9
<211>  28
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  ORF146 forward primer

<400>  9
atgggatcca atgctgcaat tgcggaga                                        28


<210>  10
<211>  23
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  ORF146 reverse primer

<400>  10
cgaggttaga acagattttg cct                                             23


<210>  11
<211>  55
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  extended ORF146 forward primer

<400>  11
atgggatcca aacgcaagaa acgtaagaaa cgcaaaaatg ctgcaattgc ggaga          55


<210>  12
<211>  768
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  DNA sequence of recombinant PK-PVPSE1gp146

<400>  12
atgggatcca aacgcaagaa acgtaagaaa cgcaaaaatg ctgcaattgc ggagattcag     60

cgtatgctga tcgaaggtgg gtttagcgtc ggcaagtctg gtgctgatgg attgtacgga    120

cccgctacaa aagccgcact gcaaaagtgc attgcacagg ctaccagtgg aaacaataaa    180

ggaggtactt tgaaactcac ccaagcacaa ctggacaaaa tcttccccgt tggtgcaagt    240

tctgggagga atgcaaaatt cctgaagccg ctcaatgacc tgtttgaaaa gacagagatt    300

aatacggtaa atcgggttgc aggattcctg tctcagattg gtgtggagtc ggcggagttc    360

cggtatgtac gtgaactcgg taacgatgcc tactttgaca agtacgacac tggtcctatt    420

gcagaaagac ttggaaacac accccagaaa gatggggatg gtgccaagta caaggggaga    480

ggtctgattc aggtgaccgg actcgcaaac tacaaggctt gcggtaaagc actcggtctt    540

gacctcgtta accaccctga gttgcttgaa cagcctgagt atgcagttgc cagcgctggt    600

tggtattggg acacgagaaa catcaacgcc gcttgcgatg ctgatgatat cgtgaaaatt    660

accaagctgg taaacggtgg tacaaatcac cttgccgagc gcacagccta ttacaaaaag    720

gcaaaatctg ttctaacctc gaagggtcat catcaccatc accattga                 768


<210>  13
<211>  254
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  amino acid sequence of recombinant PK-PVPSE1gp146

<400>  13

Met Gly Ser Lys Arg Lys Lys Arg Lys Lys Arg Lys Asn Ala Ala Ile 
1               5                   10                  15      


Ala Glu Ile Gln Arg Met Leu Ile Glu Gly Gly Phe Ser Val Gly Lys 
            20                  25                  30          


Ser Gly Ala Asp Gly Leu Tyr Gly Pro Ala Thr Lys Ala Ala Leu Gln 
        35                  40                  45              


Lys Cys Ile Ala Gln Ala Thr Ser Gly Asn Asn Lys Gly Gly Thr Leu 
    50                  55                  60                  


Lys Leu Thr Gln Ala Gln Leu Asp Lys Ile Phe Pro Val Gly Ala Ser 
65                  70                  75                  80  


Ser Gly Arg Asn Ala Lys Leu Lys Pro Leu Asn Asp Leu Phe Glu Lys 
                85                  90                  95      


Thr Glu Ile Asn Thr Val Asn Arg Val Ala Gly Phe Leu Ser Gln Ile 
            100                 105                 110         


Gly Val Glu Ser Ala Glu Phe Arg Tyr Val Arg Glu Leu Gly Asn Asp 
        115                 120                 125             


Ala Tyr Phe Asp Lys Tyr Asp Thr Gly Pro Ile Ala Glu Arg Leu Gly 
    130                 135                 140                 


Asn Thr Pro Gln Lys Asp Gly Asp Gly Ala Lys Tyr Lys Gly Arg Gly 
145                 150                 155                 160 


Leu Ile Gln Val Thr Gly Leu Ala Asn Tyr Lys Ala Cys Gly Lys Ala 
                165                 170                 175     


Leu Gly Leu Asp Leu Val Asn His Pro Glu Leu Leu Glu Gln Pro Glu 
            180                 185                 190         


Tyr Ala Val Ala Ser Ala Gly Trp Tyr Trp Asp Thr Arg Asn Ile Asn 
        195                 200                 205             


Ala Ala Cys Asp Ala Asp Asp Ile Val Lys Ile Thr Lys Leu Val Asn 
    210                 215                 220                 


Gly Gly Thr Asn His Leu Ala Glu Arg Thr Ala Tyr Tyr Lys Lys Ala 
225                 230                 235                 240 


Lys Ser Val Leu Thr Ser Lys Gly His His His His His His 
                245                 250                 


<210>  14
<211>  13
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  alpha4-helix of T4-lysozyme

<400>  14

Pro Asn Arg Ala Lys Arg Val Ile Thr Thr Phe Arg Thr 
1               5                   10              


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

<220>
<223>  Pentapeptide

<400>  15

Phe Phe Val Ala Pro 
1               5   


<210>  16
<211>  18
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Artilysin 1

<400>  16

Gly Phe Phe Ile Pro Ala Val Ile Leu Pro Ser Ile Ala Phe Leu Ile 
1               5                   10                  15      


Val Pro 
        


<210>  17
<211>  25
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Artilysin 2

<400>  17

Gly Lys Pro Gly Trp Leu Ile Lys Lys Ala Leu Val Phe Lys Lys Leu 
1               5                   10                  15      


Ile Arg Arg Pro Leu Lys Arg Leu Ala 
            20                  25  


<210>  18
<211>  19
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Parasin 1

<400>  18

Lys Gly Arg Gly Lys Gln Gly Gly Lys Val Arg Ala Lys Ala Lys Thr 
1               5                   10                  15      


Arg Ser Ser 
            


<210>  19
<211>  25
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Lycotoxin 1

<400>  19

Ile Trp Leu Thr Ala Leu Lys Phe Leu Gly Lys His Ala Ala Lys Lys 
1               5                   10                  15      


Leu Ala Lys Gln Gln Leu Ser Lys Leu 
            20                  25  


<210>  20
<211>  39
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  forward primer of alpha4-helix of T4-lysozyme

<400>  20
ttggaatggg gagcccgaac cgtgcaaaac gtgtaatca                            39


<210>  21
<211>  39
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  reverse primer of alpha4-helix of T4-lysozyme

<400>  21
cattggagga gccggtacgg aaggtggtga ttacacgtt                            39


<210>  22
<211>  39
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  forward primer of Artilysin 1

<400>  22
ttatgggctt cttcatcccg gcagtaatcc tgccctcca                            39


<210>  23
<211>  46
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  reverse primer of Artilysin 1

<400>  23
cattggagga gcccggtacg atcaggaatg cgatggaggg caggat                    46


<210>  24
<211>  47
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  forward primer of Artilysin 2

<400>  24
ttatgggcaa accgggctgg ctgatcaaaa ggcactggta ttcaaga                   47


<210>  25
<211>  57
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  reverse primer of Artilysin 2

<400>  25
cattggagga gcctgccagt ctcttcagcg gacgacggat cagtttcttg aatacca        57


<210>  26
<211>  48
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  forward primer of Parasin 1

<400>  26
ttggaatggg gagcaaaggc cgtggcaagc agggaggcaa agtacgtg                  48


<210>  27
<211>  48
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  reverse primer of Parasin 1

<400>  27
cattggagga gcctgaggaa cgggtctttg cttttgcacg tactttgc                  48


<210>  28
<211>  55
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  forward primer of Lycotoxin 1

<400>  28
ggaatgggga gcatctggct gaccgcactg aaattcctcg gcaaacacgc cgcaa          55


<210>  29
<211>  57
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  reverse primer of Lycotoxin 1

<400>  29
cattggagga gcccagtttg gataattgct gttttgccag tttctttgcg gcgtgtt        57


<210>  30
<211>  38
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  5' forward primer for tail PCR

<400>  30
ggaatgggga gctcctccaa tgctgcaatt gcggagat                             38


<210>  31
<211>  44
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  extended 5' forward primer for pentapeptide

<400>  31
atgggatcct tcttcgtagc accgggctcc tccaatgctg caat                      44


