
                                SEQUENCE LISTING

<110> NETT, Juergen

<120> PICHIA PASTORIS LOCI ENCODING ENZYMES IN
  THE PROLINE BIOSYNTHETIC PATHWAY
  

<130> GFI-MIS-00018-US

<150> 61/411,958          
<151> 2010-11-10  

<160> 6

<170> FastSEQ for Windows Version 4.0

<210> 1
<211> 2199
<212> DNA
<213> Pichia pastoris

<400> 1
tgcggaatat gccgagtcgc ctttgatgga acctgccctg tatgcaagtt ccccggagat 60
caatgtccaa ttggtaagtt ttttgattgt cttttttgtt ttgtaatttt gcgctcacat 120
gtgggccatg tactgacttt ttgcccagca attggctcct gccatcactc gtttcacatg 180
cattgcattt taaaatggtt ggatacagac acttcaaagg ggttgtgtcc aatgtgtaga 240
cagatattca ccttggagaa ctccaataac aaggaagtct tcgaagcact tagtgggagg 300
ccgtcccctt tggaggaagg cattctcgaa caggagggaa cgcccgtcta gaaaattacg 360
taacctcaat ttgtgcacgt ggatattgtt agcattgtag cagacttccg atcaacttac 420
gactatgaca tcataggtct agaggttcca aaattttgct gaaatgaaaa gactttttac 480
ccttaaagtt tattgcaaaa catgggtgaa aattgttaca ctatcgtcgt taaattggga 540
accagttccg tggtagatga agaaaccagg gaaccaagaa tcgcaactat gtctcgtatt 600
gtcgagaccc ttgttaagct gcgaaggaag ggccatcgaa ttgtcatcgt ctcctcaggt 660
gccatcgctg tgggcatgaa gagagtcaac atgaagcgga aaccaaaaaa gttacagcaa 720
gtgcaggcat tggctgctat aggacaaggc cgtttgatag gactttggga cgaccttttc 780
cgtcagttga atcagcctat tgcgcagatt ttactgacta gaacggattt ggtcgattac 840
acccagttta agaacgctga aaatacattg gaacagctta ttaaaatggg tattattcct 900
attgtcaatg agaatgacac cctatccatt caagaaatca aatttggtga caatgacacc 960
ttatccgcca taacagctgg tatgtgtcat gcagactacc tgtttttggt gactgatgtg 1020
gactgtcttt acacggataa ccctcgtacg aatccggacg ctgagccaat cgtgttagtt 1080
agaaatatga ggaatctaaa cgtcaatacc gaaagtggag gttccgccgt aggaacagga 1140
ggaatgacaa ctaaattgat cgcagctgat ttgggtgtat ctgcaggtgt tacaacgatt 1200
atttgcaaaa gtgaacatcc cgagcagatt ttggacattg tagagtacag tatccgtgct 1260
gatagagtcg aaaatgaggc taaatatctg gtcatcaacg aagaggaaac tgtggaacaa 1320
tttcaagaga tcaatcggtc agaactgagg gagttgaaca agctggacat tcctttgcat 1380
acacgtttcg ttggccacag ttttaatgct gttaataaca aagagttttg gttactccat 1440
ggactaaagg ccaacggagc cattatcatt gatccaggtt gttataaggc tatcactaga 1500
aaaaacaaag ctggtattct tccagctgga attatttccg tagagggtaa tttccatgaa 1560
tacgagtgtg ttgatgttaa ggtaggacta agagatccag atgacccaca ttcactagac 1620
cccaatgaag aactttacgt cgttggccgt gcccgttgta attaccccag caatcaaatc 1680
aacaaaatta agggtctaca aagctcgcag atcgagcagg ttctaggtta cgctgacggt 1740
gagtatgttg ttcacaggga caacttggct ttcccagtat ttgccgatcc agaactgttg 1800
gatgttgttg agagtaccct gtctgaacag gagagagaat ccaaaccaaa taaatagaat 1860
ttcacatatg ctgcttgatt atgtaattat accttgcgtt cgatggcatc gatttcctct 1920
tctgtcaatc gcgcatcgca ttaaaagtat actttttttt ttttcctata gtactattcg 1980
ccttattata aactttgcta gtatgagttc tacccccaag aaagagcctg atttgactcc 2040
taagaagagt cagcctccaa agaatagtct cggtgggggt aaaggcttta gtgaggaggg 2100
tttctcccaa ggggacttca gcgctaagca tatactaaat cgtcgcccta acaccgaagg 2160
ctcttctgtg gcttcgaacg tcatcagttc gtcatcatt                        2199

<210> 2
<211> 451
<212> PRT
<213> Pichia pastoris

<400> 2
Met Gly Glu Asn Cys Tyr Thr Ile Val Val Lys Leu Gly Thr Ser Ser
 1               5                  10                  15      
Val Val Asp Glu Glu Thr Arg Glu Pro Arg Ile Ala Thr Met Ser Arg
            20                  25                  30          
Ile Val Glu Thr Leu Val Lys Leu Arg Arg Lys Gly His Arg Ile Val
        35                  40                  45              
Ile Val Ser Ser Gly Ala Ile Ala Val Gly Met Lys Arg Val Asn Met
    50                  55                  60                  
Lys Arg Lys Pro Lys Lys Leu Gln Gln Val Gln Ala Leu Ala Ala Ile
65                  70                  75                  80  
Gly Gln Gly Arg Leu Ile Gly Leu Trp Asp Asp Leu Phe Arg Gln Leu
                85                  90                  95      
Asn Gln Pro Ile Ala Gln Ile Leu Leu Thr Arg Thr Asp Leu Val Asp
            100                 105                 110         
Tyr Thr Gln Phe Lys Asn Ala Glu Asn Thr Leu Glu Gln Leu Ile Lys
        115                 120                 125             
Met Gly Ile Ile Pro Ile Val Asn Glu Asn Asp Thr Leu Ser Ile Gln
    130                 135                 140                 
Glu Ile Lys Phe Gly Asp Asn Asp Thr Leu Ser Ala Ile Thr Ala Gly
145                 150                 155                 160 
Met Cys His Ala Asp Tyr Leu Phe Leu Val Thr Asp Val Asp Cys Leu
                165                 170                 175     
Tyr Thr Asp Asn Pro Arg Thr Asn Pro Asp Ala Glu Pro Ile Val Leu
            180                 185                 190         
Val Arg Asn Met Arg Asn Leu Asn Val Asn Thr Glu Ser Gly Gly Ser
        195                 200                 205             
Ala Val Gly Thr Gly Gly Met Thr Thr Lys Leu Ile Ala Ala Asp Leu
    210                 215                 220                 
Gly Val Ser Ala Gly Val Thr Thr Ile Ile Cys Lys Ser Glu His Pro
225                 230                 235                 240 
Glu Gln Ile Leu Asp Ile Val Glu Tyr Ser Ile Arg Ala Asp Arg Val
                245                 250                 255     
Glu Asn Glu Ala Lys Tyr Leu Val Ile Asn Glu Glu Glu Thr Val Glu
            260                 265                 270         
Gln Phe Gln Glu Ile Asn Arg Ser Glu Leu Arg Glu Leu Asn Lys Leu
        275                 280                 285             
Asp Ile Pro Leu His Thr Arg Phe Val Gly His Ser Phe Asn Ala Val
    290                 295                 300                 
Asn Asn Lys Glu Phe Trp Leu Leu His Gly Leu Lys Ala Asn Gly Ala
305                 310                 315                 320 
Ile Ile Ile Asp Pro Gly Cys Tyr Lys Ala Ile Thr Arg Lys Asn Lys
                325                 330                 335     
Ala Gly Ile Leu Pro Ala Gly Ile Ile Ser Val Glu Gly Asn Phe His
            340                 345                 350         
Glu Tyr Glu Cys Val Asp Val Lys Val Gly Leu Arg Asp Pro Asp Asp
        355                 360                 365             
Pro His Ser Leu Asp Pro Asn Glu Glu Leu Tyr Val Val Gly Arg Ala
    370                 375                 380                 
Arg Cys Asn Tyr Pro Ser Asn Gln Ile Asn Lys Ile Lys Gly Leu Gln
385                 390                 395                 400 
Ser Ser Gln Ile Glu Gln Val Leu Gly Tyr Ala Asp Gly Glu Tyr Val
                405                 410                 415     
Val His Arg Asp Asn Leu Ala Phe Pro Val Phe Ala Asp Pro Glu Leu
            420                 425                 430         
Leu Asp Val Val Glu Ser Thr Leu Ser Glu Gln Glu Arg Glu Ser Lys
        435                 440                 445             
Pro Asn Lys
    450     


<210> 3
<211> 2451
<212> DNA
<213> Pichia pastoris

<400> 3
atcaatctgt aatcgcagat ggcacctcca taccggaagt atccgacaga ttgtccaaac 60
ttgatgtatc cagaatgtat gccataggta caaactgctg cagcatcagc actgctaacg 120
gggcagtgga gctattctct aaacatacaa accttccatt gatcgtttac cccaattctg 180
gggagagata tgacaaaact gaaaaaaaat ggctccctgg tgaatgcgac caaaaaataa 240
ctgacatcgt agtcaattgg ctgcaactga atgtaaagat cattggagga tgttgtcgga 300
ccaatcccca ctttattcgt caacttagag atatagtcga caacagtagc agaagtggat 360
gaagatatga catccgacat ttaacatctc agtaattttt ttttctcgac ctgacatctc 420
atgaatagca tgaatagggt atgacgaaat gacgcactcg aattattttc atctacctct 480
ttctacccta tttatataaa atggctgacg ttattgctaa gaatgcccat ttggcatcca 540
cagtgttgaa aactttgact gacaatgaga ggtctgatgc cttggtaaag gttgagcgtg 600
ccttgaaact gaacaaggat cgaatcttgg aggccaactt ggtggacatg gagacagcaa 660
agaagaataa tctaagtgac tcgttgatta aaagattgga tttatccaag aacgacaagt 720
ttgattccat gtgccagggt gtgttggatg ttgcccaact ggaagaccct ttaaacaaga 780
tcactttagc cacaaagatt gatgacggac tgaatttgta tcgattaact tgccctgttg 840
gagtgttgtt gatcatattc gaaagtcgtc cagaagttat tgccaatatc actgctttgg 900
caatcaaatc tggtaatgct gccattctga aaggtggtaa ggagagtttc aacactttca 960
aggcattgtc tgacgtaatc aatgagactc tgactaaaga gacgaaagta ccagagaact 1020
ccattcaact tatttccacc agaagcgatg tcagtgactt gttgagtcag gacaagtaca 1080
tcgatctggt gattcccaga ggctccaacg cattagttcg taacatcaag gacaatacca 1140
agattcctgt cttgggacat gctgatggaa tttgctctat ctacgtggat gagtttgccc 1200
aacttgataa agctaccaag attcttatcg actctaagac caactatcca gctggttgta 1260
atgctgtgga gaaatgcctc atcgacgaga ctttagtaaa gtcgaactta gacgataagg 1320
ttgttcctct attgaaaagt ttgatagaag ctggtatcac tctccacatc gatcctgaac 1380
tgcttaaatt gttacctcaa gagatttctg ctaacgaaaa gactgttgca gcaacggaag 1440
acgacttttc cagagagttc ctgtcctttg accttgccgt caaggccgtt aagaatgttc 1500
aagaggctat tctccacata aatgagcatt cttcaaaaca tacggactgt atcataaccg 1560
agaacaaaga aaatgccgat agattcctac aaggtatcga ctcctcagga gtttactgga 1620
attgttctac cagatttgct gatggtttcc gttatggatt tggtgctgaa gttggtatct 1680
ccaccaacaa aattcattct cgtggacctg tgggactaga gggattagtt tgctactact 1740
accaaatgcg tggagatggc cacattgctg gcgattacgc tggtgccgga ggggttcggg 1800
ttttcaagca tgagaaaatc gaccccaatt cagtagtgtt gaactaaagt aatgccatgt 1860
ttagatcgta tatagcggca actatgacat gtttactctc attcctcgac cgaatgtgtc 1920
tgccaagata agactaataa gtttgctgca tgtttatgca tattttactt tatcaccatg 1980
gaaacataca tattcttgta gctttggcgt tctctgcccg tcatcgaaac agtcgtttat 2040
gcaaacagac ttaacctgtt ctcgaagcgg tctccagcta tcatcatgcc aatacttgct 2100
gtacctgtag gagcttgtcg tgggttatct cttccatcat gaaatgacta acagaacccc 2160
catctaccat agaactaata ggaatgcacc tgaaacgaaa gtggttatca gtcattactt 2220
atcagtaaaa ggactacctg taattaatgc ttcaaatgac tagttgagtg catcatcatg 2280
tacagtaaaa aaaagaaaag ttgcgctagt aaattaaaaa gaagcgtggc cctgcttgcc 2340
atgtggcccc cacttgactt tctcctttat tggcaccagt aaaaccggta cacaaattcc 2400
actccagcaa ctgtcagaca gtgacatatt ttttattgca ccacttactg a          2451

<210> 4
<211> 448
<212> PRT
<213> Pichia pastoris

<400> 4
Met Ala Asp Val Ile Ala Lys Asn Ala His Leu Ala Ser Thr Val Leu
 1               5                  10                  15      
Lys Thr Leu Thr Asp Asn Glu Arg Ser Asp Ala Leu Val Lys Val Glu
            20                  25                  30          
Arg Ala Leu Lys Leu Asn Lys Asp Arg Ile Leu Glu Ala Asn Leu Val
        35                  40                  45              
Asp Met Glu Thr Ala Lys Lys Asn Asn Leu Ser Asp Ser Leu Ile Lys
    50                  55                  60                  
Arg Leu Asp Leu Ser Lys Asn Asp Lys Phe Asp Ser Met Cys Gln Gly
65                  70                  75                  80  
Val Leu Asp Val Ala Gln Leu Glu Asp Pro Leu Asn Lys Ile Thr Leu
                85                  90                  95      
Ala Thr Lys Ile Asp Asp Gly Leu Asn Leu Tyr Arg Leu Thr Cys Pro
            100                 105                 110         
Val Gly Val Leu Leu Ile Ile Phe Glu Ser Arg Pro Glu Val Ile Ala
        115                 120                 125             
Asn Ile Thr Ala Leu Ala Ile Lys Ser Gly Asn Ala Ala Ile Leu Lys
    130                 135                 140                 
Gly Gly Lys Glu Ser Phe Asn Thr Phe Lys Ala Leu Ser Asp Val Ile
145                 150                 155                 160 
Asn Glu Thr Leu Thr Lys Glu Thr Lys Val Pro Glu Asn Ser Ile Gln
                165                 170                 175     
Leu Ile Ser Thr Arg Ser Asp Val Ser Asp Leu Leu Ser Gln Asp Lys
            180                 185                 190         
Tyr Ile Asp Leu Val Ile Pro Arg Gly Ser Asn Ala Leu Val Arg Asn
        195                 200                 205             
Ile Lys Asp Asn Thr Lys Ile Pro Val Leu Gly His Ala Asp Gly Ile
    210                 215                 220                 
Cys Ser Ile Tyr Val Asp Glu Phe Ala Gln Leu Asp Lys Ala Thr Lys
225                 230                 235                 240 
Ile Leu Ile Asp Ser Lys Thr Asn Tyr Pro Ala Gly Cys Asn Ala Val
                245                 250                 255     
Glu Lys Cys Leu Ile Asp Glu Thr Leu Val Lys Ser Asn Leu Asp Asp
            260                 265                 270         
Lys Val Val Pro Leu Leu Lys Ser Leu Ile Glu Ala Gly Ile Thr Leu
        275                 280                 285             
His Ile Asp Pro Glu Leu Leu Lys Leu Leu Pro Gln Glu Ile Ser Ala
    290                 295                 300                 
Asn Glu Lys Thr Val Ala Ala Thr Glu Asp Asp Phe Ser Arg Glu Phe
305                 310                 315                 320 
Leu Ser Phe Asp Leu Ala Val Lys Ala Val Lys Asn Val Gln Glu Ala
                325                 330                 335     
Ile Leu His Ile Asn Glu His Ser Ser Lys His Thr Asp Cys Ile Ile
            340                 345                 350         
Thr Glu Asn Lys Glu Asn Ala Asp Arg Phe Leu Gln Gly Ile Asp Ser
        355                 360                 365             
Ser Gly Val Tyr Trp Asn Cys Ser Thr Arg Phe Ala Asp Gly Phe Arg
    370                 375                 380                 
Tyr Gly Phe Gly Ala Glu Val Gly Ile Ser Thr Asn Lys Ile His Ser
385                 390                 395                 400 
Arg Gly Pro Val Gly Leu Glu Gly Leu Val Cys Tyr Tyr Tyr Gln Met
                405                 410                 415     
Arg Gly Asp Gly His Ile Ala Gly Asp Tyr Ala Gly Ala Gly Gly Val
            420                 425                 430         
Arg Val Phe Lys His Glu Lys Ile Asp Pro Asn Ser Val Val Leu Asn
        435                 440                 445             


<210> 5
<211> 1672
<212> DNA
<213> Pichia pastoris

<400> 5
tcatatatgt gacgaacaca acatcatcag tagacaaagt gaacttttat tcatcctctt 60
cttcctcgga agcttcattg gatgaaaaca aacaaatcgc agaatccaaa gagaccactc 120
cacaattgga ccgcttgtcc tttgtcgaat tcagaacggg aagacctaac cttgaggaag 180
ttctcaatag agatatcagc gagtccaatg gaaccgtctc tgtattaact tgtggccatc 240
cacatatgtg tgatcacgtc agagatgcca tagctcgtaa caccccaaac tataagaaga 300
gagtagatta ctacgaatca gtgcaagtgt ggtaatctga cttgcccatc ccattgcctt 360
atataatact atcattatca ttaagatcta tatatgtgtt tcgcgcctcc catgtatcta 420
ccagatcggc atttgaattt gcaatatttc actaacttct gtcttatctc ccactttagt 480
ttgcttaatc actaataaag atgtcttcaa ttggtaacga atatacaatc actatcctag 540
ggtgcggagt gatgggaacc gcagttttat ctgctatttt gaactcagag ttctcccctt 600
atccctcaag aattatctgc tgcacaggct cccagaagtc cgctgaaaaa ttggccagca 660
cttattccaa gattgaaact agctacggat ctgagcaaaa catcagagcc gtcaaagagt 720
cagatatcat aattctaggg tgcaagccgt tctattgtga acaaatcatg gaacaggtca 780
aagagggctt acacgatcaa ttgctaattt cacttttggc tggctggacg ttaacccaac 840
tagaaaggta ctctaaatat gtagcaaggg taatgaccaa taccccggcc aaatttggat 900
gtggaatggc ttgtgtatct ctatcttcgc acgccgagca atacgaagat ctagtactca 960
agctggttgg cccagtgggc aaggctatta gattgcccga aaagaatatg gatgccgcta 1020
cagctcttgt aggttcaggt cctgcattct gcttgttgat gatggaatct ctcatcgacg 1080
gagccgtcca aaagggttta ccatttgcta ttgctaaaga tgcagctgct aaagttatgg 1140
aaggcacagc aaggatggtt ctcgagactg gagaacatcc tgccgcacta aaaagtgctg 1200
tgtgcacccc aggaggtacc actattggtg gtcttatggc catggaagac cgaggagtca 1260
gaagtggtat cgctcgaggt gtggaagaag ctgccaacat tgccactagc ttaggaaaga 1320
aatagggtaa attcactgac tgcctctttc tttcaaatca gacatgatat tctcaaattc 1380
ttgctctcct aactcattga tgatttcttc cacgtcgtcc cgatctcctc gtaaaatctg 1440
gtcttgaatc actttggcac tgattgattc ttgctgtgat tctttctctt tttcctcatt 1500
caaaagagaa tacaggaaaa agttgaacca aatctccatg aataagaaac taaagatcaa 1560
cccattgcaa agatagtacg atgaacccac atagcagtag aatatcagga tcagaaacag 1620
tgaaacccga actgggacaa ttgatctgaa gtagacgaca tttccttcta at         1672

<210> 6
<211> 274
<212> PRT
<213> Pichia pastoris

<400> 6
Met Ser Ser Ile Gly Asn Glu Tyr Thr Ile Thr Ile Leu Gly Cys Gly
 1               5                  10                  15      
Val Met Gly Thr Ala Val Leu Ser Ala Ile Leu Asn Ser Glu Phe Ser
            20                  25                  30          
Pro Tyr Pro Ser Arg Ile Ile Cys Cys Thr Gly Ser Gln Lys Ser Ala
        35                  40                  45              
Glu Lys Leu Ala Ser Thr Tyr Ser Lys Ile Glu Thr Ser Tyr Gly Ser
    50                  55                  60                  
Glu Gln Asn Ile Arg Ala Val Lys Glu Ser Asp Ile Ile Ile Leu Gly
65                  70                  75                  80  
Cys Lys Pro Phe Tyr Cys Glu Gln Ile Met Glu Gln Val Lys Glu Gly
                85                  90                  95      
Leu His Asp Gln Leu Leu Ile Ser Leu Leu Ala Gly Trp Thr Leu Thr
            100                 105                 110         
Gln Leu Glu Arg Tyr Ser Lys Tyr Val Ala Arg Val Met Thr Asn Thr
        115                 120                 125             
Pro Ala Lys Phe Gly Cys Gly Met Ala Cys Val Ser Leu Ser Ser His
    130                 135                 140                 
Ala Glu Gln Tyr Glu Asp Leu Val Leu Lys Leu Val Gly Pro Val Gly
145                 150                 155                 160 
Lys Ala Ile Arg Leu Pro Glu Lys Asn Met Asp Ala Ala Thr Ala Leu
                165                 170                 175     
Val Gly Ser Gly Pro Ala Phe Cys Leu Leu Met Met Glu Ser Leu Ile
            180                 185                 190         
Asp Gly Ala Val Gln Lys Gly Leu Pro Phe Ala Ile Ala Lys Asp Ala
        195                 200                 205             
Ala Ala Lys Val Met Glu Gly Thr Ala Arg Met Val Leu Glu Thr Gly
    210                 215                 220                 
Glu His Pro Ala Ala Leu Lys Ser Ala Val Cys Thr Pro Gly Gly Thr
225                 230                 235                 240 
Thr Ile Gly Gly Leu Met Ala Met Glu Asp Arg Gly Val Arg Ser Gly
                245                 250                 255     
Ile Ala Arg Gly Val Glu Glu Ala Ala Asn Ile Ala Thr Ser Leu Gly
            260                 265                 270         
Lys Lys
        

