               SEQUENCE LISTING

<110> Xbrane Biopharma AB
      STADA Arzneimittel AG

<120> METHOD OF PRODUCING A RECOMBINANT PROTEIN IN A HOST CELL WHICH HAS A DISABLED RHAMNOSE METABOLISM AS WELL AS EXPRESSION VECTORS, HOST CELLS AND RECOMBINANT PROTEINS THEREOF 

<130> S12481WO/RN

<150> EP 20201096.3
<151> 2020-10-09

<160> 21

<170> BiSSAP 1.3.6

<210> 1
<211> 4673
<212> DNA
<213> Artificial Sequence


<220> 
<223> KTXHIS

<400> 1
atctttctgc gaattgagat gacgccactg gctgggcgtc atcccggttt cccgggtaaa       60

caccaccgaa aaatagttac tatcttcaaa gccacattcg gtcgaaatat cactgattaa      120

caggcggcta tgctggagaa gatattgcgc atgacacact ctgacctgtc gcagatattg      180

attgatggtc attccagtct gctggcgaaa ttgctgacgc aaaacgcgct cactgcacga      240

tgcctcatca caaaatttat ccagcgcaaa gggacttttc aggctagccg ccagccgggt      300

aatcagctta tccagcaacg tttcgctgga tgttggcggc aacgaatcac tggtgtaacg      360

atggcgattc agcaacatca ccaactgccc gaacagcaac tcagccattt cgttagcaaa      420

cggcacatgc tgactacttt catgctcaag ctgaccaata acctgccgcg cctgcgccat      480

ccccatgcta cctaagcgcc agtgtggttg ccctgcgctg gcgttaaatc ccggaatcgc      540

cccctgccag tcaagattca gcttcagacg ctccgggcaa taaataatat tctgcaaaac      600

cagatcgtta acggaagcgt aggagtgttt atcatcagca tgaatgtaaa agagatcgcc      660

acgggtaatg cgataagggc gatcgttgag tacatgcagg ccattaccgc gccagacaat      720

caccagctca caaaaatcat gtgtatgttc agcaaagaca tcttgcggat aacggtcagc      780

cacagcgact gcctgctggt cgctggcaaa aaaatcatct ttgagaagtt ttaactgatg      840

cgccaccgtg gctacctcgg ccagagaacg aagttgatta ttcgcaatat ggcgtacaaa      900

tacgttgaga agattcgcgt tattgcagaa agccatcccg tccctggcga atatcacgcg      960

gtgaccagtt aaactctcgg cgaaaaagcg tcgaaaagtg gttactgtcg ctgaatccac     1020

agcgataggc gatgtcagta acgctggcct cgctgtggcg tagcagatgt cgggctttca     1080

tcagtcgcag gcggttcagg tatcgctgag gcgtcagtcc cgtttgctgc ttaagctgcc     1140

gatgtagcgt acgcagtgaa agagaaaatt gatccgccac ggcatcccaa ttcacctcat     1200

cggcaaaatg gtcctccagc caggccagaa gcaagttgag acgtgatgcg ctgttttcca     1260

ggttctcctg caaactgctt ttacgcagca agagcagtaa ttgcataaac aagatctcgc     1320

gactggcggt cgagggtaaa tcattttccc cttcctgctg ttccatctgt gcaaccagct     1380

gtcgcacctg ctgcaatacg ctgtggttaa cgcgccagtg agacggatac tgcccatcca     1440

gctcttgtgg cagcaactga ttcagcccgg cgagaaactg aaatcgatcc ggcgagcgat     1500

acagcacatt ggtcagacac agattatcgg tatgttcata cagatgccga tcatgatcgc     1560

gtacgaaaca gaccgtgcca ccggtgatgg tatagggctg cccattaaac acatgaatac     1620

ccgtgccatg ttcgacaatc acaatttcat gaaaatcatg atgatgttca ggaaaatccg     1680

cctgcgggag ccggggttct atcgccacgg acgcgttacc agacggaaaa aaatccacac     1740

tatgtaatac ggtcatactg gcctcctgat gtcgtcaaca cggcgaaata gtaatcacga     1800

ggtcaggttc ttaccttaaa ttttcgacgg aaaaccacgt aaaaaacgtc gatttttcaa     1860

gatacagcgt gaattttcag gaaatgcggt gagcatcaca tcaccacaat tcagcaaatt     1920

gtgaacatca tcacgttcat ctttccctgg ttgccaatgg cccattttct tgtcagtaac     1980

gagaaggtcg cgaatccagg cgctttttag actggtcgta atgaaattca ggaggaattc     2040

catatggcgg ccgcactcga gcaccaccac caccaccact gatgaagacg gccgcctagg     2100

ggtaccgagc tcggatcctc tagagtcgac ctgcaggcat gcaagcttgg ctgttttggc     2160

ggatgagaga agattttcag cctgatacag attaaatcag aacgcagaag cggtctgata     2220

aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat gccgaactca     2280

gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc cccatgcgag agtagggaac     2340

tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg     2400

ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt     2460

tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggcatca     2520

aattaagcag aaggccatcc tgacggatgg cctttttgcg tttctacaaa ctcttttgtt     2580

tatttttcta aatacattca aatatgtatc cgctcatgag actaggcttc cgcgccctca     2640

tccgaaaggg cgtattcata tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa     2700

taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg     2760

ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg     2820

gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag     2880

gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga     2940

cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct     3000

ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc     3060

tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg     3120

gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc     3180

tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca     3240

ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag     3300

ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct     3360

ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc     3420

accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga     3480

tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca     3540

cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat     3600

taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttag     3660

aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca     3720

tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg     3780

atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt     3840

aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa     3900

tccggtgaga atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca     3960

ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc     4020

tgagcgagac gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc     4080

aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct     4140

tctaatacct ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca tgcatcatca     4200

ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt     4260

ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac     4320

tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta     4380

tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa tcgcggccta     4440

gagcaagacg tttcccgttg aatatggctc atactcttcc tttttcaata ttattgaagc     4500

atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa     4560

caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt     4620

attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tct            4673


<210> 2
<211> 767
<212> DNA
<213> Artificial Sequence


<220> 
<223> frame shift mutation

<400> 2
gttggccccg ctccacgcca gtagcgactc gtcccagtcg tcgctattga tattgaccag      60

ttgcgtggtc gtggcgttgg tatattccca gttcatcttg ccggtcaggc gataactgaa     120

gtaatccggc atcagcagag cgtgagcaat gtgtggaata agttcaggtt gttgctccgt     180

cagcgcacgc aactgataaa gcgtattgaa gggcagaaac tggatgccgc tacgttgata     240

aatatcgcgt ttgccgagtt gttgttgtgc ctgcgccatt aggccattgg tgcggctatc     300

gcgataagca acgggcaggc ccacacgctg accctgttgg tcgagcagca caaagtccac     360

gccccaggta tcaatcccaa tgctatcgcg atacgaatcc cttcctcgca caccttgttt     420

aatccaaggc gaatggcact ttccaggcta tccacatccc aggtgacata gccgttctga     480

ctatgcagcc cattgttaaa acgatggatt tcgcgcagcg tcaggctgcg gcattcacgc     540

tcgtaacgcg ccagcatcac gcgcccactg gatgcgccga gatcgacggc gacacaattg     600

cgaaaggtca taatgtgatc ctgctgaatt tcattacgac cagtctaaaa agcgcctgaa     660

ttcgcgacct tctcgttact gacaggaaaa tgggccattg gcaaccaggg aaagatgaac     720

gtgatgatgt tcacaatttg ctgaattgtg gtgatgtgat gctcacc                   767


<210> 3
<211> 214
<212> PRT
<213> Artificial Sequence


<220> 
<223> Ranibizumab light chain

<400> 3
Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 
1               5                   10                  15      
Asp Arg Val Thr Ile Thr Cys Ser Ala Ser Gln Asp Ile Ser Asn Tyr 
            20                  25                  30          
Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Val Leu Ile 
        35                  40                  45              
Tyr Phe Thr Ser Ser Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly 
    50                  55                  60                  
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 
65                  70                  75                  80  
Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Ser Thr Val Pro Trp 
                85                  90                  95      
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala 
            100                 105                 110         
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 
        115                 120                 125             
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 
    130                 135                 140                 
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 
145                 150                 155                 160 
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 
                165                 170                 175     
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 
            180                 185                 190         
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 
        195                 200                 205             
Phe Asn Arg Gly Glu Cys 
    210                 

<210> 4
<211> 231
<212> PRT
<213> Artificial Sequence


<220> 
<223> Ranibizumab heavy chain

<400> 4
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 
1               5                   10                  15      
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Asp Phe Thr His Tyr 
            20                  25                  30          
Gly Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 
        35                  40                  45              
Gly Trp Ile Asn Thr Tyr Thr Gly Glu Pro Thr Tyr Ala Ala Asp Phe 
    50                  55                  60                  
Lys Arg Arg Phe Thr Phe Ser Leu Asp Thr Ser Lys Ser Thr Ala Tyr 
65                  70                  75                  80  
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 
                85                  90                  95      
Ala Lys Tyr Pro Tyr Tyr Tyr Gly Thr Ser His Trp Tyr Phe Asp Val 
            100                 105                 110         
Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly 
        115                 120                 125             
Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly 
    130                 135                 140                 
Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val 
145                 150                 155                 160 
Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe 
                165                 170                 175     
Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val 
            180                 185                 190         
Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val 
        195                 200                 205             
Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys 
    210                 215                 220                 
Ser Cys Asp Lys Thr His Leu 
225                 230     

<210> 5
<211> 645
<212> DNA
<213> Artificial Sequence


<220> 
<223> ranibizumab light chain

<400> 5
gatattcagc tgacccagag cccgagcagc ctgagcgcaa gcgttggtga tcgtgttacc      60

attacctgta gcgcaagcca ggatattagc aattatctga attggtatca gcagaaaccg     120

ggtaaagcac cgaaagtgct gatctatttt accagcagcc tgcatagcgg tgttccgagc     180

cgttttagcg gtagcggtag tggcaccgat tttaccctga ccattagcag cctgcagccg     240

gaagattttg caacctatta ttgtcagcag tatagcaccg ttccgtggac ctttggtcag     300

ggcaccaaag ttgaaattaa acgtaccgtt gcagcaccga gcgtttttat ctttccgcct     360

agtgatgaac agctgaaaag cggcaccgca agcgttgttt gtctgctgaa taacttttat     420

ccgcgtgaag caaaagttca gtggaaagtt gataatgcac tgcagagcgg taatagccaa     480

gaaagcgtta ccgaacagga tagcaaagat agcacctata gcctgagcag caccctgacc     540

ctgagcaaag cagattatga aaaacacaaa gtgtatgcct gcgaagttac ccatcagggt     600

ctgagcagtc cggttaccaa aagttttaat cgtggtgaat gctaa                     645


<210> 6
<211> 696
<212> DNA
<213> Artificial Sequence


<220> 
<223> ranibizumab heavy chain

<400> 6
gaagttcagc tggttgaaag cggtggtggt ctggttcagc ctggtggtag cctgcgtctg      60

agctgtgcag caagcggtta tgattttacc cattatggta tgaattgggt tcgtcaggca     120

ccgggtaaag gtctggaatg ggttggttgg attaatacct ataccggtga accgacctat     180

gcagcagatt ttaaacgtcg ttttaccttt agcctggata ccagcaaaag caccgcatat     240

ctgcagatga atagcctgcg tgcagaggat accgcagtgt attattgtgc aaaatatccg     300

tattattacg gcaccagcca ttggtatttc gatgtttggg gtcagggcac cctggttacc     360

gttagcagcg caagcaccaa aggtccgagc gtttttccgc tggcaccgag cagcaaaagt     420

accagcggtg gcaccgcagc actgggttgt ctggttaaag attattttcc ggaaccggtt     480

accgtgagct ggaatagcgg tgcactgacc agcggtgttc atacctttcc ggcagttctg     540

cagagcagcg gtctgtatag cctgagcagc gttgttaccg ttccgagcag cagcctgggc     600

acccagacct atatttgtaa tgttaatcat aaaccgagca ataccaaagt ggataaaaaa     660

gtggaaccga aaagctgcga taaaacccat ctgtaa                               696


<210> 7
<211> 66
<212> DNA
<213> Artificial Sequence


<220> 
<223> PelB signal peptide - DNA

<400> 7
atgaaatatc tgctgccgac cgcagcagcg ggtctgctgc tgctggcagc acagcctgca     60

atggca                                                                66


<210> 8
<211> 119
<212> DNA
<213> Artificial Sequence


<220> 
<223> rhaBAD promoter

<400> 8
caccacaatt cagcaaattg tgaacatcat cacgttcatc tttccctggt tgccaatggc     60

ccattttctt gtcagtaacg agaaggtcgc gaatccaggc gctttttaga ctggtcgta     119


<210> 9
<211> 936
<212> DNA
<213> Artificial Sequence


<220> 
<223> RhaR transcription activator

<400> 9
atggctttct gcaataacgc gaatcttctc aacgtatttg tacgccatat tgcgaataat      60

caacttcgtt ctctggccga ggtagccacg gtggcgcatc agttaaaact tctcaaagat     120

gatttttttg ccagcgacca gcaggcagtc gctgtggctg accgttatcc gcaagatgtc     180

tttgctgaac atacacatga tttttgtgag ctggtgattg tctggcgcgg taatggcctg     240

catgtactca acgatcgccc ttatcgcatt acccgtggcg atctctttta cattcatgct     300

gatgataaac actcctacgc ttccgttaac gatctggttt tgcagaatat tatttattgc     360

ccggagcgtc tgaagctgaa tcttgactgg cagggggcga ttccgggatt taacgccagc     420

gcagggcaac cacactggcg cttaggtagc atggggatgg cgcaggcgcg gcaggttatt     480

ggtcagcttg agcatgaaag tagtcagcat gtgccgtttg ctaacgaaat ggctgagttg     540

ctgttcgggc agttggtgat gttgctgaat cgccatcgtt acaccagtga ttcgttgccg     600

ccaacatcca gcgaaacgtt gctggataag ctgattaccc ggctggcggc tagcctgaaa     660

agtccctttg cgctggataa attttgtgat gaggcatcgt gcagtgagcg cgttttgcgt     720

cagcaatttc gccagcagac tggaatgacc atcaatcaat atctgcgaca ggtcagagtg     780

tgtcatgcgc aatatcttct ccagcatagc cgcctgttaa tcagtgatat ttcgaccgaa     840

tgtggctttg aagatagtaa ctatttttcg gtggtgttta cccgggaaac cgggatgacg     900

cccagccagt ggcgtcatct caattcgcag aaagat                               936


<210> 10
<211> 36
<212> DNA
<213> Artificial Sequence


<220> 
<223> RhaR extended

<400> 10
agacgaaagg gcctcgtgat acgcctattt ttatag                               36


<210> 11
<211> 837
<212> DNA
<213> Artificial Sequence


<220> 
<223> RhaS transcription activator

<400> 11
atgaccgtat tacatagtgt ggattttttt ccgtctggta acgcgtccgt ggcgatagaa      60

ccccggctcc cgcaggcgga ttttcctgaa catcatcatg attttcatga aattgtgatt     120

gtcgaacatg gcacgggtat tcatgtgttt aatgggcagc cctataccat caccggtggc     180

acggtctgtt tcgtacgcga tcatgatcgg catctgtatg aacataccga taatctgtgt     240

ctgaccaatg tgctgtatcg ctcgccggat cgatttcagt ttctcgccgg gctgaatcag     300

ttgctgccac aagagctgga tgggcagtat ccgtctcact ggcgcgttaa ccacagcgta     360

ttgcagcagg tgcgacagct ggttgcacag atggaacagc aggaagggga aaatgattta     420

ccctcgaccg ccagtcgcga gatcttgttt atgcaattac tgctcttgct gcgtaaaagc     480

agtttgcagg agaacctgga aaacagcgca tcacgtctca acttgcttct ggcctggctg     540

gaggaccatt ttgccgatga ggtgaattgg gatgccgtgg cggatcaatt ttctctttca     600

ctgcgtacgc tacatcggca gcttaagcag caaacgggac tgacgcctca gcgatacctg     660

aaccgcctgc gactgatgaa agcccgacat ctgctacgcc acagcgaggc cagcgttact     720

gacatcgcct atcgctgtgg attcagcgac agtaaccact tttcgacgct ttttcgccga     780

gagtttaact ggtcaccgcg tgatattcgc cagggacggg atggctttct gcaataa        837


<210> 12
<211> 816
<212> DNA
<213> Artificial Sequence


<220> 
<223> KanR

<400> 12
atgagccata ttcaacggga aacgtcttgc tctaggccgc gattaaattc caacatggat      60

gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc     120

tatcgattgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc     180

gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct     240

cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg     300

atccccggga aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt     360

gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct     420

tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg     480

gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa     540

gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca     600

cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc     660

ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct     720

ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa     780

ttgcagtttc atttgatgct cgatgagttt ttctaa                               816


<210> 13
<211> 105
<212> DNA
<213> Artificial Sequence


<220> 
<223> AmpR promoter

<400> 13
cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga     60

caataaccct gataaatgct tcaataatat tgaaaaagga agagt                    105


<210> 14
<211> 87
<212> DNA
<213> Artificial Sequence


<220> 
<223> rrnB T1 terminator

<400> 14
caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg     60

gtgaacgctc tcctgagtag gacaaat                                         87


<210> 15
<211> 28
<212> DNA
<213> Artificial Sequence


<220> 
<223> rrnB T2 terminator

<400> 15
agaaggccat cctgacggat ggcctttt                                       28


<210> 16
<211> 589
<212> DNA
<213> Artificial Sequence


<220> 
<223> pMB1 origin of replication

<400> 16
tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg      60

gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg     120

ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag     180

cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc     240

caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa     300

ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg     360

taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc     420

taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac     480

cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg     540

tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaa                 589


<210> 17
<211> 6088
<212> DNA
<213> Artificial Sequence


<220> 
<223> KTXHIS-pelbLC-pelbHC

<400> 17
atctttctgc gaattgagat gacgccactg gctgggcgtc atcccggttt cccgggtaaa       60

caccaccgaa aaatagttac tatcttcaaa gccacattcg gtcgaaatat cactgattaa      120

caggcggcta tgctggagaa gatattgcgc atgacacact ctgacctgtc gcagatattg      180

attgatggtc attccagtct gctggcgaaa ttgctgacgc aaaacgcgct cactgcacga      240

tgcctcatca caaaatttat ccagcgcaaa gggacttttc aggctagccg ccagccgggt      300

aatcagctta tccagcaacg tttcgctgga tgttggcggc aacgaatcac tggtgtaacg      360

atggcgattc agcaacatca ccaactgccc gaacagcaac tcagccattt cgttagcaaa      420

cggcacatgc tgactacttt catgctcaag ctgaccaata acctgccgcg cctgcgccat      480

ccccatgcta cctaagcgcc agtgtggttg ccctgcgctg gcgttaaatc ccggaatcgc      540

cccctgccag tcaagattca gcttcagacg ctccgggcaa taaataatat tctgcaaaac      600

cagatcgtta acggaagcgt aggagtgttt atcatcagca tgaatgtaaa agagatcgcc      660

acgggtaatg cgataagggc gatcgttgag tacatgcagg ccattaccgc gccagacaat      720

caccagctca caaaaatcat gtgtatgttc agcaaagaca tcttgcggat aacggtcagc      780

cacagcgact gcctgctggt cgctggcaaa aaaatcatct ttgagaagtt ttaactgatg      840

cgccaccgtg gctacctcgg ccagagaacg aagttgatta ttcgcaatat ggcgtacaaa      900

tacgttgaga agattcgcgt tattgcagaa agccatcccg tccctggcga atatcacgcg      960

gtgaccagtt aaactctcgg cgaaaaagcg tcgaaaagtg gttactgtcg ctgaatccac     1020

agcgataggc gatgtcagta acgctggcct cgctgtggcg tagcagatgt cgggctttca     1080

tcagtcgcag gcggttcagg tatcgctgag gcgtcagtcc cgtttgctgc ttaagctgcc     1140

gatgtagcgt acgcagtgaa agagaaaatt gatccgccac ggcatcccaa ttcacctcat     1200

cggcaaaatg gtcctccagc caggccagaa gcaagttgag acgtgatgcg ctgttttcca     1260

ggttctcctg caaactgctt ttacgcagca agagcagtaa ttgcataaac aagatctcgc     1320

gactggcggt cgagggtaaa tcattttccc cttcctgctg ttccatctgt gcaaccagct     1380

gtcgcacctg ctgcaatacg ctgtggttaa cgcgccagtg agacggatac tgcccatcca     1440

gctcttgtgg cagcaactga ttcagcccgg cgagaaactg aaatcgatcc ggcgagcgat     1500

acagcacatt ggtcagacac agattatcgg tatgttcata cagatgccga tcatgatcgc     1560

gtacgaaaca gaccgtgcca ccggtgatgg tatagggctg cccattaaac acatgaatac     1620

ccgtgccatg ttcgacaatc acaatttcat gaaaatcatg atgatgttca ggaaaatccg     1680

cctgcgggag ccggggttct atcgccacgg acgcgttacc agacggaaaa aaatccacac     1740

tatgtaatac ggtcatactg gcctcctgat gtcgtcaaca cggcgaaata gtaatcacga     1800

ggtcaggttc ttaccttaaa ttttcgacgg aaaaccacgt aaaaaacgtc gatttttcaa     1860

gatacagcgt gaattttcag gaaatgcggt gagcatcaca tcaccacaat tcagcaaatt     1920

gtgaacatca tcacgttcat ctttccctgg ttgccaatgg cccattttct tgtcagtaac     1980

gagaaggtcg cgaatccagg cgctttttag actggtcgta atgaaattca ggaggaattc     2040

ctaatgaaat atctgctgcc gaccgcagca gcgggtctgc tgctgctggc agcacagcct     2100

gcaatggcag atattcagct gacccagagc ccgagcagcc tgagcgcaag cgttggtgat     2160

cgtgttacca ttacctgtag cgcaagccag gatattagca attatctgaa ttggtatcag     2220

cagaaaccgg gtaaagcacc gaaagtgctg atctatttta ccagcagcct gcatagcggt     2280

gttccgagcc gttttagcgg tagcggtagt ggcaccgatt ttaccctgac cattagcagc     2340

ctgcagccgg aagattttgc aacctattat tgtcagcagt atagcaccgt tccgtggacc     2400

tttggtcagg gcaccaaagt tgaaattaaa cgtaccgttg cagcaccgag cgtttttatc     2460

tttccgccta gtgatgaaca gctgaaaagc ggcaccgcaa gcgttgtttg tctgctgaat     2520

aacttttatc cgcgtgaagc aaaagttcag tggaaagttg ataatgcact gcagagcggt     2580

aatagccaag aaagcgttac cgaacaggat agcaaagata gcacctatag cctgagcagc     2640

accctgaccc tgagcaaagc agattatgaa aaacacaaag tgtatgcctg cgaagttacc     2700

catcagggtc tgagcagtcc ggttaccaaa agttttaatc gtggtgaatg ctaataatct     2760

agaactgtta tcgatatcag gaggattagc atatgaaata tctgctgccg accgcagcag     2820

cgggtctgct gctgctggca gcacagcctg caatggcaga agttcagctg gttgaaagcg     2880

gtggtggtct ggttcagcct ggtggtagcc tgcgtctgag ctgtgcagca agcggttatg     2940

attttaccca ttatggtatg aattgggttc gtcaggcacc gggtaaaggt ctggaatggg     3000

ttggttggat taatacctat accggtgaac cgacctatgc agcagatttt aaacgtcgtt     3060

ttacctttag cctggatacc agcaaaagca ccgcatatct gcagatgaat agcctgcgtg     3120

cagaggatac cgcagtgtat tattgtgcaa aatatccgta ttattacggc accagccatt     3180

ggtatttcga tgtttggggt cagggcaccc tggttaccgt tagcagcgca agcaccaaag     3240

gtccgagcgt ttttccgctg gcaccgagca gcaaaagtac cagcggtggc accgcagcac     3300

tgggttgtct ggttaaagat tattttccgg aaccggttac cgtgagctgg aatagcggtg     3360

cactgaccag cggtgttcat acctttccgg cagttctgca gagcagcggt ctgtatagcc     3420

tgagcagcgt tgttaccgtt ccgagcagca gcctgggcac ccagacctat atttgtaatg     3480

ttaatcataa accgagcaat accaaagtgg ataaaaaagt ggaaccgaaa agctgcgata     3540

aaacccatct gtaataaaag cttggctgtt ttggcggatg agagaagatt ttcagcctga     3600

tacagattaa atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta     3660

gcgcggtggt cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg     3720

gtagtgtggg gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag     3780

gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg     3840

agtaggacaa atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg     3900

cgggcaggac gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg     3960

gatggccttt ttgcgtttct acaaactctt ttgtttattt ttctaaatac attcaaatat     4020

gtatccgctc atgagactag gcttccgcgc cctcatccga aagggcgtat tcatatatgc     4080

ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt     4140

cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact     4200

caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag     4260

caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata     4320

ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc     4380

cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg     4440

ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc     4500

tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg     4560

gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc     4620

ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga     4680

ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg     4740

gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa     4800

aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg     4860

tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt     4920

ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat     4980

tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct     5040

aaagtatata tgagtaaact tggtctgaca gttagaaaaa ctcatcgagc atcaaatgaa     5100

actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta     5160

atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg     5220

cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt     5280

tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat     5340

gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg     5400

catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc     5460

tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg     5520

catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc     5580

cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg     5640

tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat     5700

tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca     5760

atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata     5820

aatcagcatc catgttggaa tttaatcgcg gcctagagca agacgtttcc cgttgaatat     5880

ggctcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga     5940

gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc     6000

cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa     6060

ataggcgtat cacgaggccc tttcgtct                                        6088


<210> 18
<211> 22
<212> PRT
<213> Erwinia


<220> 
<223> PelB signal peptide - peptide

<400> 18
Met Lys Tyr Leu Leu Pro Thr Ala Ala Ala Gly Leu Leu Leu Leu Ala 
1               5                   10                  15      
Ala Gln Pro Ala Met Ala 
            20          

<210> 19
<211> 32
<212> DNA
<213> Artificial Sequence


<220> 
<223> SBamHI1680

<400> 19
gtacgctagg atcctgtggc agcaactgat tc                                   32


<210> 20
<211> 32
<212> DNA
<213> Artificial Sequence


<220> 
<223> BSalI1681

<400> 20
gtagataggt cgacagcgat cgtcattggg at                                   32


<210> 21
<211> 180
<212> DNA
<213> Artificial Sequence


<220> 
<223> Figure 2 - Multiple cloning site

<400> 21
gagaaggtcg cgaatccagg cgctttttag actggtcgta atgaaattca ggaggaattc     60

catatggcgg ccgcactcga gcaccaccac caccaccact gatgaagacg gccgcctagg    120

ggtaccgagc tcggatcctc tagagtcgac ctgcaggcat gcaagcttgg ctgttttggc    180


