                         SEQUENCE LISTING

<110>  Merck Sharp and Dohme
 
<120>  CRZ1 Mutant Fungal Cells

<130>  23341

<160>  9     

<170>  PatentIn version 3.5

<210>  1
<211>  676
<212>  PRT
<213>  Saccharomyces cerevisiae

<400>  1

Met Ser Phe Ser Asn Gly Asn Met Ala Ser Tyr Met Thr Ser Ser Asn 
1               5                   10                  15      


Gly Glu Glu Gln Ser Ile Asn Asn Lys Asn Asp Ile Asp Asp Asn Ser 
            20                  25                  30          


Ala Tyr Arg Arg Asn Asn Phe Arg Asn Ser Ser Asn Ser Gly Ser His 
        35                  40                  45              


Thr Phe Gln Leu Ser Asp Leu Asp Leu Asp Val Asp Met Arg Met Asp 
    50                  55                  60                  


Ser Ala Asn Ser Ser Glu Lys Ile Ser Lys Asn Leu Ser Ser Gly Ile 
65                  70                  75                  80  


Pro Asp Ser Phe Asp Ser Asn Val Asn Ser Leu Leu Ser Pro Ser Ser 
                85                  90                  95      


Gly Ser Tyr Ser Ala Asp Leu Asn Tyr Gln Ser Leu Tyr Lys Pro Asp 
            100                 105                 110         


Leu Pro Gln Gln Gln Leu Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln 
        115                 120                 125             


Gln Gln Gln Gln Gln Gln Gln Gln Lys Gln Thr Pro Thr Leu Lys Val 
    130                 135                 140                 


Glu Gln Ser Asp Thr Phe Gln Trp Asp Asp Ile Leu Thr Pro Ala Asp 
145                 150                 155                 160 


Asn Gln His Arg Pro Ser Leu Thr Asn Gln Phe Leu Ser Pro Arg Ser 
                165                 170                 175     


Asn Tyr Asp Gly Thr Thr Arg Ser Ser Gly Ile Asp Ser Asn Tyr Ser 
            180                 185                 190         


Asp Thr Glu Ser Asn Tyr His Thr Pro Tyr Leu Tyr Pro Gln Asp Leu 
        195                 200                 205             


Val Ser Ser Pro Ala Met Ser His Leu Thr Ala Asn Asn Asp Asp Phe 
    210                 215                 220                 


Asp Asp Leu Leu Ser Val Ala Ser Met Asn Ser Asn Tyr Leu Leu Pro 
225                 230                 235                 240 


Val Asn Ser His Gly Tyr Lys His Ile Ser Asn Leu Asp Glu Leu Asp 
                245                 250                 255     


Asp Leu Leu Ser Leu Thr Tyr Ser Asp Asn Asn Leu Leu Ser Ala Ser 
            260                 265                 270         


Asn Asn Ser Asp Phe Asn Asn Ser Asn Asn Gly Ile Ile Asn Thr Ala 
        275                 280                 285             


Asp Thr Gln Asn Ser Thr Ile Ala Ile Asn Lys Ser Lys Val Gly Thr 
    290                 295                 300                 


Asn Gln Lys Met Leu Leu Thr Ile Pro Thr Ser Ser Thr Pro Ser Pro 
305                 310                 315                 320 


Ser Thr His Ala Ala Pro Val Thr Pro Ile Ile Ser Ile Gln Glu Phe 
                325                 330                 335     


Asn Glu Gly His Phe Pro Val Lys Asn Glu Asp Asp Gly Thr Leu Gln 
            340                 345                 350         


Leu Lys Val Arg Asp Asn Glu Ser Tyr Ser Ala Thr Asn Asn Asn Asn 
        355                 360                 365             


Leu Leu Arg Pro Asp Asp Asn Asp Tyr Asn Asn Glu Ala Leu Ser Asp 
    370                 375                 380                 


Ile Asp Arg Ser Phe Glu Asp Ile Ile Asn Gly Arg Lys Leu Lys Leu 
385                 390                 395                 400 


Lys Lys Ser Arg Arg Arg Ser Ser Gln Thr Ser Asn Asn Ser Phe Thr 
                405                 410                 415     


Ser Arg Arg Ser Ser Arg Ser Arg Ser Ile Ser Pro Asp Glu Lys Ala 
            420                 425                 430         


Lys Ser Ile Ser Ala Asn Arg Glu Lys Leu Leu Glu Met Ala Asp Leu 
        435                 440                 445             


Leu Pro Ser Ser Glu Asn Asp Asn Asn Arg Glu Arg Tyr Asp Asn Asp 
    450                 455                 460                 


Ser Lys Thr Ser Tyr Asn Thr Ile Asn Ser Ser Asn Phe Asn Glu Asp 
465                 470                 475                 480 


Asn Asn Asn Asn Asn Leu Leu Thr Ser Lys Pro Lys Ile Glu Ser Gly 
                485                 490                 495     


Ile Val Asn Ile Lys Asn Glu Leu Asp Asp Thr Ser Lys Asp Leu Gly 
            500                 505                 510         


Ile Leu Leu Asp Ile Asp Ser Leu Gly Gln Phe Glu Gln Lys Val Gly 
        515                 520                 525             


Phe Lys Asn Asp Asp Asn His Glu Asn Asn Asp Asn Gly Thr Phe Ser 
    530                 535                 540                 


Val Lys Lys Asn Asp Asn Leu Glu Lys Leu Asp Ser Val Thr Asn Asn 
545                 550                 555                 560 


Arg Lys Asn Pro Ala Asn Phe Ala Cys Asp Val Cys Gly Lys Lys Phe 
                565                 570                 575     


Thr Arg Pro Tyr Asn Leu Lys Ser His Leu Arg Thr His Thr Asn Glu 
            580                 585                 590         


Arg Pro Phe Ile Cys Ser Ile Cys Gly Lys Ala Phe Ala Arg Gln His 
        595                 600                 605             


Asp Arg Lys Arg His Glu Asp Leu His Thr Gly Lys Lys Arg Tyr Val 
    610                 615                 620                 


Cys Gly Gly Lys Leu Lys Asp Gly Lys Pro Trp Gly Cys Gly Lys Lys 
625                 630                 635                 640 


Phe Ala Arg Ser Asp Ala Leu Gly Arg His Phe Lys Thr Glu Ser Gly 
                645                 650                 655     


Arg Arg Cys Ile Thr Pro Leu Tyr Glu Glu Ala Arg Gln Glu Lys Ser 
            660                 665                 670         


Gly Gln Glu Ser 
        675     


<210>  2
<211>  1557
<212>  DNA
<213>  Pichia pastoris


<220>
<221>  CDS
<222>  (1)..(1557)

<400>  2
atg gca gac caa cgg ctt gag gat gag ttt gat atc tcc aga tac tta         48
Met Ala Asp Gln Arg Leu Glu Asp Glu Phe Asp Ile Ser Arg Tyr Leu           
1               5                   10                  15                

tct att tct cct atc gag tca gct tca atc gaa gaa tca atc aac ggt         96
Ser Ile Ser Pro Ile Glu Ser Ala Ser Ile Glu Glu Ser Ile Asn Gly           
            20                  25                  30                    

tta atg agt agt tgg att cct ccg gct aag ggt gag att aga gat tca        144
Leu Met Ser Ser Trp Ile Pro Pro Ala Lys Gly Glu Ile Arg Asp Ser           
        35                  40                  45                        

ctt cct cca aac gct tct ttt gaa gct aca gac agt ttt tca acc agt        192
Leu Pro Pro Asn Ala Ser Phe Glu Ala Thr Asp Ser Phe Ser Thr Ser           
    50                  55                  60                            

tca tac cag gaa att ata cca gca cag gtg aaa ata aaa ctg gag ttt        240
Ser Tyr Gln Glu Ile Ile Pro Ala Gln Val Lys Ile Lys Leu Glu Phe           
65                  70                  75                  80            

gat aat gac cag cag cct gtt ttc tat caa gaa tcg caa cca gtt tat        288
Asp Asn Asp Gln Gln Pro Val Phe Tyr Gln Glu Ser Gln Pro Val Tyr           
                85                  90                  95                

gat aag cat tta acc gtc aat gat cag gaa acc aga agc gcc caa gac        336
Asp Lys His Leu Thr Val Asn Asp Gln Glu Thr Arg Ser Ala Gln Asp           
            100                 105                 110                   

ttc aac caa tac ttg aat gct gat gcc gta tcg agg acc aac tcc atc        384
Phe Asn Gln Tyr Leu Asn Ala Asp Ala Val Ser Arg Thr Asn Ser Ile           
        115                 120                 125                       

tcc aac tta tcg gag ctg tca act cat tcc cat att acc cct cca acg        432
Ser Asn Leu Ser Glu Leu Ser Thr His Ser His Ile Thr Pro Pro Thr           
    130                 135                 140                           

cta ctt cat gat caa gcc tca ttg tct cct gct ctc tta tct atg aac        480
Leu Leu His Asp Gln Ala Ser Leu Ser Pro Ala Leu Leu Ser Met Asn           
145                 150                 155                 160           

agt gat gaa aga aac gaa ctc aat ctg gaa aca cta cag cta gat caa        528
Ser Asp Glu Arg Asn Glu Leu Asn Leu Glu Thr Leu Gln Leu Asp Gln           
                165                 170                 175               

acg tca cag cct tac gtg aat cag ata aaa acg gag gca gct tac gaa        576
Thr Ser Gln Pro Tyr Val Asn Gln Ile Lys Thr Glu Ala Ala Tyr Glu           
            180                 185                 190                   

gag ctt tca gag tta cac cac aga tta gaa aga ctc act gag aca aat        624
Glu Leu Ser Glu Leu His His Arg Leu Glu Arg Leu Thr Glu Thr Asn           
        195                 200                 205                       

tta att cat caa gac cag ctt caa ctc gaa caa caa gag caa caa aat        672
Leu Ile His Gln Asp Gln Leu Gln Leu Glu Gln Gln Glu Gln Gln Asn           
    210                 215                 220                           

cag act cct cat act ctc agt cct cct ata caa ctt cag act ccc ata        720
Gln Thr Pro His Thr Leu Ser Pro Pro Ile Gln Leu Gln Thr Pro Ile           
225                 230                 235                 240           

atc aaa gtc ttg caa gcc cca aat gat ata gca gca aat acc ccg tct        768
Ile Lys Val Leu Gln Ala Pro Asn Asp Ile Ala Ala Asn Thr Pro Ser           
                245                 250                 255               

ctt ttt tct caa tct aac cat tca tct cca tat aac aca ccc aaa cat        816
Leu Phe Ser Gln Ser Asn His Ser Ser Pro Tyr Asn Thr Pro Lys His           
            260                 265                 270                   

tcc agg tca aac tcg ttg agt tca aat gac aga caa cat gat att cca        864
Ser Arg Ser Asn Ser Leu Ser Ser Asn Asp Arg Gln His Asp Ile Pro           
        275                 280                 285                       

caa ata tcc tca gtt tta gac acg tct tcg ttt ttg gta cct gga gat        912
Gln Ile Ser Ser Val Leu Asp Thr Ser Ser Phe Leu Val Pro Gly Asp           
    290                 295                 300                           

cag ttt caa gca atg aga gaa ggt aga cag agg agg aaa tcc gag tcc        960
Gln Phe Gln Ala Met Arg Glu Gly Arg Gln Arg Arg Lys Ser Glu Ser           
305                 310                 315                 320           

aac tct aga aac tcg aaa gaa cgt tct aaa tct agg gaa ccg cca aag       1008
Asn Ser Arg Asn Ser Lys Glu Arg Ser Lys Ser Arg Glu Pro Pro Lys           
                325                 330                 335               

tcc agg tct aga tct aga gac agt gca aca gat cat cat atg gaa gtt       1056
Ser Arg Ser Arg Ser Arg Asp Ser Ala Thr Asp His His Met Glu Val           
            340                 345                 350                   

atg agc aga gaa aag act ctt gag ttg gca gct tct cag cca agc tcc       1104
Met Ser Arg Glu Lys Thr Leu Glu Leu Ala Ala Ser Gln Pro Ser Ser           
        355                 360                 365                       

aag acg ccg caa aag aac cct tcc atc tat gct tgc tcg ctc tgc tcc       1152
Lys Thr Pro Gln Lys Asn Pro Ser Ile Tyr Ala Cys Ser Leu Cys Ser           
    370                 375                 380                           

aag aga ttc aca aga cca tat aat ttg aag tct cac ctt cgc acg cac       1200
Lys Arg Phe Thr Arg Pro Tyr Asn Leu Lys Ser His Leu Arg Thr His           
385                 390                 395                 400           

gct gat gaa agg cct ttc cag tgt tca ata tgc ggg aag gca ttt gct       1248
Ala Asp Glu Arg Pro Phe Gln Cys Ser Ile Cys Gly Lys Ala Phe Ala           
                405                 410                 415               

cgt tct cac gac aga aag cgt cat gag gat ctg cat agt ggt gaa cga       1296
Arg Ser His Asp Arg Lys Arg His Glu Asp Leu His Ser Gly Glu Arg           
            420                 425                 430                   

aag tat tgc tgc aaa ggt gtt ttg tct gac gga gta act aca tgg ggc       1344
Lys Tyr Cys Cys Lys Gly Val Leu Ser Asp Gly Val Thr Thr Trp Gly           
        435                 440                 445                       

tgt gaa aaa aga ttt gcc cga aca gat gcg ctg ggt aga cat ttc aaa       1392
Cys Glu Lys Arg Phe Ala Arg Thr Asp Ala Leu Gly Arg His Phe Lys           
    450                 455                 460                           

act gaa tgt ggt aaa ctg tgt atc aag ccg ctg atg gat gaa cta aag       1440
Thr Glu Cys Gly Lys Leu Cys Ile Lys Pro Leu Met Asp Glu Leu Lys           
465                 470                 475                 480           

agg gag gaa gct tac agg agg aat gaa cca gta aca gaa atg aat gac       1488
Arg Glu Glu Ala Tyr Arg Arg Asn Glu Pro Val Thr Glu Met Asn Asp           
                485                 490                 495               

gag ctt tac tcc caa tct gtc caa gat ata ttt agc tct cag cgg ctt       1536
Glu Leu Tyr Ser Gln Ser Val Gln Asp Ile Phe Ser Ser Gln Arg Leu           
            500                 505                 510                   

ggt cag aac ata gat gac tga                                           1557
Gly Gln Asn Ile Asp Asp                                                   
        515                                                               


<210>  3
<211>  518
<212>  PRT
<213>  Pichia pastoris

<400>  3

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


Ser Ile Ser Pro Ile Glu Ser Ala Ser Ile Glu Glu Ser Ile Asn Gly 
            20                  25                  30          


Leu Met Ser Ser Trp Ile Pro Pro Ala Lys Gly Glu Ile Arg Asp Ser 
        35                  40                  45              


Leu Pro Pro Asn Ala Ser Phe Glu Ala Thr Asp Ser Phe Ser Thr Ser 
    50                  55                  60                  


Ser Tyr Gln Glu Ile Ile Pro Ala Gln Val Lys Ile Lys Leu Glu Phe 
65                  70                  75                  80  


Asp Asn Asp Gln Gln Pro Val Phe Tyr Gln Glu Ser Gln Pro Val Tyr 
                85                  90                  95      


Asp Lys His Leu Thr Val Asn Asp Gln Glu Thr Arg Ser Ala Gln Asp 
            100                 105                 110         


Phe Asn Gln Tyr Leu Asn Ala Asp Ala Val Ser Arg Thr Asn Ser Ile 
        115                 120                 125             


Ser Asn Leu Ser Glu Leu Ser Thr His Ser His Ile Thr Pro Pro Thr 
    130                 135                 140                 


Leu Leu His Asp Gln Ala Ser Leu Ser Pro Ala Leu Leu Ser Met Asn 
145                 150                 155                 160 


Ser Asp Glu Arg Asn Glu Leu Asn Leu Glu Thr Leu Gln Leu Asp Gln 
                165                 170                 175     


Thr Ser Gln Pro Tyr Val Asn Gln Ile Lys Thr Glu Ala Ala Tyr Glu 
            180                 185                 190         


Glu Leu Ser Glu Leu His His Arg Leu Glu Arg Leu Thr Glu Thr Asn 
        195                 200                 205             


Leu Ile His Gln Asp Gln Leu Gln Leu Glu Gln Gln Glu Gln Gln Asn 
    210                 215                 220                 


Gln Thr Pro His Thr Leu Ser Pro Pro Ile Gln Leu Gln Thr Pro Ile 
225                 230                 235                 240 


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


Leu Phe Ser Gln Ser Asn His Ser Ser Pro Tyr Asn Thr Pro Lys His 
            260                 265                 270         


Ser Arg Ser Asn Ser Leu Ser Ser Asn Asp Arg Gln His Asp Ile Pro 
        275                 280                 285             


Gln Ile Ser Ser Val Leu Asp Thr Ser Ser Phe Leu Val Pro Gly Asp 
    290                 295                 300                 


Gln Phe Gln Ala Met Arg Glu Gly Arg Gln Arg Arg Lys Ser Glu Ser 
305                 310                 315                 320 


Asn Ser Arg Asn Ser Lys Glu Arg Ser Lys Ser Arg Glu Pro Pro Lys 
                325                 330                 335     


Ser Arg Ser Arg Ser Arg Asp Ser Ala Thr Asp His His Met Glu Val 
            340                 345                 350         


Met Ser Arg Glu Lys Thr Leu Glu Leu Ala Ala Ser Gln Pro Ser Ser 
        355                 360                 365             


Lys Thr Pro Gln Lys Asn Pro Ser Ile Tyr Ala Cys Ser Leu Cys Ser 
    370                 375                 380                 


Lys Arg Phe Thr Arg Pro Tyr Asn Leu Lys Ser His Leu Arg Thr His 
385                 390                 395                 400 


Ala Asp Glu Arg Pro Phe Gln Cys Ser Ile Cys Gly Lys Ala Phe Ala 
                405                 410                 415     


Arg Ser His Asp Arg Lys Arg His Glu Asp Leu His Ser Gly Glu Arg 
            420                 425                 430         


Lys Tyr Cys Cys Lys Gly Val Leu Ser Asp Gly Val Thr Thr Trp Gly 
        435                 440                 445             


Cys Glu Lys Arg Phe Ala Arg Thr Asp Ala Leu Gly Arg His Phe Lys 
    450                 455                 460                 


Thr Glu Cys Gly Lys Leu Cys Ile Lys Pro Leu Met Asp Glu Leu Lys 
465                 470                 475                 480 


Arg Glu Glu Ala Tyr Arg Arg Asn Glu Pro Val Thr Glu Met Asn Asp 
                485                 490                 495     


Glu Leu Tyr Ser Gln Ser Val Gln Asp Ile Phe Ser Ser Gln Arg Leu 
            500                 505                 510         


Gly Gln Asn Ile Asp Asp 
        515             


<210>  4
<211>  19
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Pichia pastoris

<400>  4
gtatgcgata tagtgtgga                                                    19


<210>  5
<211>  19
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Pichia pastoris

<400>  5
tggggagaag gtaccgaag                                                    19


<210>  6
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  primer

<400>  6
cactacgcgt actgtgagcc                                                   20


<210>  7
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Pichia pastoris

<400>  7
aggatatcaa acccgaccag                                                   20


<210>  8
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Pichia pastoris

<400>  8
gacacatgcg aaatgtcctg                                                   20


<210>  9
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Pichia pastoris

<400>  9
ttgagttggc agcttctcag                                                   20


