                         SEQUENCE LISTING

<110>  E.I. du Pont de Nemours & Co., Inc.
       Zhu, Quinn
       Macool, Daniel
       Xue, Zhixiong
 
<120>  EXPRESSION OF CYTOSOLIC MALIC ENZYME IN TRANSGENIC YARROWIA TO 
       INCREASE LIPID PRODUCTION

<130>  CL5392USNP

<150>  61/619,574
<151>  2004-04-03

<160>  32    

<170>  PatentIn version 3.5

<210>  1
<211>  629
<212>  PRT
<213>  Yarrowia lipolytica


<220>
<221>  MISC_FEATURE
<222>  (1)..(629)
<223>  GenBank Accession No. XP_504112 (malic enzyme)

<400>  1

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


Ala Leu Gly Pro Thr Ala Ala Ala Arg Asn Met Ser Ser Ser Ser Pro 
            20                  25                  30          


Ser Ser Phe Glu Tyr Ser Ser Tyr Val Lys Gly Thr Arg Glu Ile Gly 
        35                  40                  45              


His Arg Lys Ala Pro Thr Thr Arg Leu Ser Val Glu Gly Pro Ile Tyr 
    50                  55                  60                  


Val Gly Phe Asp Gly Ile Arg Leu Leu Asn Leu Pro His Leu Asn Lys 
65                  70                  75                  80  


Gly Ser Gly Phe Pro Leu Asn Glu Arg Arg Glu Phe Arg Leu Ser Gly 
                85                  90                  95      


Leu Leu Pro Ser Ala Glu Ala Thr Leu Glu Glu Gln Val Asp Arg Ala 
            100                 105                 110         


Tyr Gln Gln Phe Lys Lys Cys Gly Thr Pro Leu Ala Lys Asn Gly Phe 
        115                 120                 125             


Cys Thr Ser Leu Lys Phe Gln Asn Glu Val Leu Tyr Tyr Ala Leu Leu 
    130                 135                 140                 


Leu Lys His Val Lys Glu Val Phe Pro Ile Ile Tyr Thr Pro Thr Gln 
145                 150                 155                 160 


Gly Glu Ala Ile Glu Gln Tyr Ser Arg Leu Phe Arg Arg Pro Glu Gly 
                165                 170                 175     


Cys Phe Leu Asp Ile Thr Ser Pro Tyr Asp Val Glu Glu Arg Leu Gly 
            180                 185                 190         


Ala Phe Gly Asp His Asp Asp Ile Asp Tyr Ile Val Val Thr Asp Ser 
        195                 200                 205             


Glu Gly Ile Leu Gly Ile Gly Asp Gln Gly Val Gly Gly Ile Gly Ile 
    210                 215                 220                 


Ser Ile Ala Lys Leu Ala Leu Met Thr Leu Cys Ala Gly Val Asn Pro 
225                 230                 235                 240 


Ser Arg Val Ile Pro Val Val Leu Asp Thr Gly Thr Asn Asn Gln Glu 
                245                 250                 255     


Leu Leu His Asp Pro Leu Tyr Leu Gly Arg Arg Met Pro Arg Val Arg 
            260                 265                 270         


Gly Lys Gln Tyr Asp Asp Phe Ile Asp Asn Phe Val Gln Ser Ala Arg 
        275                 280                 285             


Arg Leu Tyr Pro Lys Ala Val Ile His Phe Glu Asp Phe Gly Leu Ala 
    290                 295                 300                 


Asn Ala His Lys Ile Leu Asp Lys Tyr Arg Pro Glu Ile Pro Cys Phe 
305                 310                 315                 320 


Asn Asp Asp Ile Gln Gly Thr Gly Ala Val Thr Leu Ala Ser Ile Thr 
                325                 330                 335     


Ala Ala Leu Lys Val Leu Gly Lys Asn Ile Thr Asp Thr Arg Ile Leu 
            340                 345                 350         


Val Tyr Gly Ala Gly Ser Ala Gly Met Gly Ile Ala Glu Gln Val Tyr 
        355                 360                 365             


Asp Asn Leu Val Ala Gln Gly Leu Asp Asp Lys Thr Ala Arg Gln Asn 
    370                 375                 380                 


Ile Phe Leu Met Asp Arg Pro Gly Leu Leu Thr Thr Ala Leu Thr Asp 
385                 390                 395                 400 


Glu Gln Met Ser Asp Val Gln Lys Pro Phe Ala Lys Asp Lys Ala Asn 
                405                 410                 415     


Tyr Glu Gly Val Asp Thr Lys Thr Leu Glu His Val Val Ala Ala Val 
            420                 425                 430         


Lys Pro His Ile Leu Ile Gly Cys Ser Thr Gln Pro Gly Ala Phe Asn 
        435                 440                 445             


Glu Lys Val Val Lys Glu Met Leu Lys His Thr Pro Arg Pro Ile Ile 
    450                 455                 460                 


Leu Pro Leu Ser Asn Pro Thr Arg Leu His Glu Ala Val Pro Ala Asp 
465                 470                 475                 480 


Leu Tyr Lys Trp Thr Asp Gly Lys Ala Leu Val Ala Thr Gly Ser Pro 
                485                 490                 495     


Phe Asp Pro Val Asn Gly Lys Glu Thr Ser Glu Asn Asn Asn Cys Phe 
            500                 505                 510         


Val Phe Pro Gly Ile Gly Leu Gly Ala Ile Leu Ser Arg Ser Lys Leu 
        515                 520                 525             


Ile Thr Asn Thr Met Ile Ala Ala Ala Ile Glu Cys Leu Ala Glu Gln 
    530                 535                 540                 


Ala Pro Ile Leu Lys Asn His Asp Glu Gly Val Leu Pro Asp Val Ala 
545                 550                 555                 560 


Leu Ile Gln Ile Ile Ser Ala Arg Val Ala Thr Ala Val Val Leu Gln 
                565                 570                 575     


Ala Lys Ala Glu Gly Leu Ala Thr Val Glu Glu Glu Leu Lys Pro Gly 
            580                 585                 590         


Thr Lys Glu His Val Gln Ile Pro Asp Asn Phe Asp Glu Cys Leu Ala 
        595                 600                 605             


Trp Val Glu Thr Gln Met Trp Arg Pro Val Tyr Arg Pro Leu Ile His 
    610                 615                 620                 


Val Arg Asp Tyr Asp 
625                 


<210>  2
<211>  669
<212>  PRT
<213>  Saccharomyces cerevisiae


<220>
<221>  MISC_FEATURE
<222>  (1)..(669)
<223>  GenBank Accession No. EDN59877 (malic enzyme)

<400>  2

Met Leu Arg Thr Arg Leu Ser Val Ser Val Ala Ala Arg Ser Gln Leu 
1               5                   10                  15      


Thr Arg Ser Leu Thr Ala Ser Arg Thr Ala Pro Leu Arg Arg Trp Pro 
            20                  25                  30          


Ile Gln Gln Ser Arg Leu Tyr Ser Ser Asn Thr Arg Ser His Lys Ala 
        35                  40                  45              


Thr Thr Thr Arg Glu Asn Thr Phe Gln Lys Pro Tyr Ser Asp Glu Glu 
    50                  55                  60                  


Val Thr Lys Thr Pro Val Gly Ser Arg Ala Arg Lys Ile Phe Glu Ala 
65                  70                  75                  80  


Pro His Pro His Ala Thr Arg Leu Thr Val Glu Gly Ala Ile Glu Cys 
                85                  90                  95      


Pro Leu Glu Ser Phe Gln Leu Leu Asn Ser Pro Leu Phe Asn Lys Gly 
            100                 105                 110         


Ser Ala Phe Thr Gln Glu Glu Arg Glu Ala Phe Asn Leu Glu Ala Leu 
        115                 120                 125             


Leu Pro Pro Gln Val Asn Thr Leu Asp Glu Gln Leu Glu Arg Ser Tyr 
    130                 135                 140                 


Lys Gln Leu Cys Tyr Leu Lys Thr Pro Leu Ala Lys Asn Asp Phe Met 
145                 150                 155                 160 


Thr Ser Leu Arg Val Gln Asn Lys Val Leu Tyr Phe Ala Leu Ile Arg 
                165                 170                 175     


Lys His Ile Lys Glu Leu Val Pro Ile Ile Tyr Thr Pro Thr Glu Gly 
            180                 185                 190         


Asp Ala Ile Ala Ala Tyr Ser His Arg Phe Arg Lys Pro Glu Gly Val 
        195                 200                 205             


Phe Leu Asp Ile Thr Glu Pro Asp Ser Ile Glu Arg Arg Leu Ala Thr 
    210                 215                 220                 


Tyr Gly Gly Asp Lys Asp Val Asp Tyr Ile Val Val Ser Asp Ser Glu 
225                 230                 235                 240 


Gly Ile Leu Gly Ile Gly Asp Gln Gly Ile Gly Gly Val Arg Ile Ala 
                245                 250                 255     


Ile Ser Lys Leu Ala Leu Met Thr Leu Cys Gly Gly Ile His Pro Gly 
            260                 265                 270         


Arg Val Leu Pro Val Cys Leu Asp Val Gly Thr Asn Asn Lys Lys Leu 
        275                 280                 285             


Ala Arg Asp Glu Leu Tyr Met Gly Asn Lys Phe Ser Arg Ile Arg Gly 
    290                 295                 300                 


Lys Gln Tyr Asp Asp Phe Leu Glu Lys Phe Ile Lys Ala Val Lys Lys 
305                 310                 315                 320 


Val Tyr Pro Ser Ala Val Leu His Phe Glu Asp Phe Gly Val Lys Asn 
                325                 330                 335     


Ala Arg Arg Leu Leu Glu Lys Tyr Arg Tyr Glu Leu Pro Ser Phe Asn 
            340                 345                 350         


Asp Asp Ile Gln Gly Thr Gly Ala Val Val Met Ala Ser Leu Ile Ala 
        355                 360                 365             


Ala Leu Lys His Thr Asn Arg Asp Leu Lys Asp Thr Arg Val Leu Ile 
    370                 375                 380                 


Tyr Gly Ala Gly Ser Ala Gly Leu Gly Ile Ala Asp Gln Ile Val Asn 
385                 390                 395                 400 


His Met Val Thr His Gly Val Asp Lys Glu Glu Ala Arg Lys Lys Ile 
                405                 410                 415     


Phe Leu Met Asp Arg Arg Gly Leu Ile Leu Gln Ser Tyr Glu Ala Asn 
            420                 425                 430         


Ser Thr Pro Ala Gln His Val Tyr Ala Lys Ser Asp Ala Glu Trp Ala 
        435                 440                 445             


Gly Ile Asn Thr Arg Ser Leu His Asp Val Val Glu Asn Val Lys Pro 
    450                 455                 460                 


Thr Cys Leu Val Gly Cys Ser Thr Gln Ala Gly Ala Phe Thr Gln Asp 
465                 470                 475                 480 


Val Val Glu Glu Met His Lys His Asn Pro Arg Pro Ile Ile Phe Pro 
                485                 490                 495     


Leu Ser Asn Pro Thr Arg Leu His Glu Ala Val Pro Ala Asp Leu Met 
            500                 505                 510         


Lys Trp Thr Asn Asn Asn Ala Leu Val Ala Thr Gly Ser Pro Phe Pro 
        515                 520                 525             


Pro Val Asp Gly Tyr Arg Ile Ser Glu Asn Asn Asn Cys Tyr Ser Phe 
    530                 535                 540                 


Pro Gly Ile Gly Leu Gly Ala Val Leu Ser Arg Ala Thr Thr Ile Thr 
545                 550                 555                 560 


Asp Lys Met Ile Ser Ala Ala Val Asp Gln Leu Ala Glu Leu Ser Pro 
                565                 570                 575     


Leu Arg Glu Gly Asp Ser Arg Pro Gly Leu Leu Pro Gly Leu Asp Thr 
            580                 585                 590         


Ile Thr Asn Thr Ser Ala Arg Leu Ala Thr Ala Val Ile Leu Gln Ala 
        595                 600                 605             


Leu Glu Glu Gly Thr Ala Arg Ile Glu Gln Glu Gln Val Pro Gly Gly 
    610                 615                 620                 


Ala Pro Gly Glu Thr Val Lys Val Pro Arg Asp Phe Asp Glu Cys Leu 
625                 630                 635                 640 


Gln Trp Val Lys Ala Gln Met Trp Glu Pro Val Tyr Arg Pro Met Ile 
                645                 650                 655     


Lys Val Gln His Asp Pro Ser Val His Thr Asn Gln Leu 
            660                 665                 


<210>  3
<211>  565
<212>  PRT
<213>  Schizosaccharomyces pombe


<220>
<221>  MISC_FEATURE
<222>  (1)..(565)
<223>  GenBank Accession No. NP_587760 (malic enzyme)

<400>  3

Met Pro Ala Gly Thr Lys Glu Gln Ile Glu Cys Pro Leu Lys Gly Val 
1               5                   10                  15      


Thr Leu Leu Asn Ser Pro Arg Tyr Asn Lys Asp Thr Ala Phe Thr Pro 
            20                  25                  30          


Glu Glu Arg Gln Lys Phe Glu Ile Ser Ser Arg Leu Pro Pro Ile Val 
        35                  40                  45              


Glu Thr Leu Gln Gln Gln Val Asp Arg Cys Tyr Asp Gln Tyr Lys Ala 
    50                  55                  60                  


Ile Gly Asp Glu Pro Leu Gln Lys Asn Leu Tyr Leu Ser Gln Leu Ser 
65                  70                  75                  80  


Val Thr Asn Gln Thr Leu Phe Tyr Ala Leu Ile Ser Gln His Leu Ile 
                85                  90                  95      


Glu Met Ile Pro Ile Ile Tyr Thr Pro Thr Glu Gly Asp Ala Ile Lys 
            100                 105                 110         


Gln Phe Ser Asp Ile Tyr Arg Tyr Pro Glu Gly Cys Tyr Leu Asp Ile 
        115                 120                 125             


Asp His Asn Asp Leu Ser Tyr Ile Lys Gln Gln Leu Ser Glu Phe Gly 
    130                 135                 140                 


Lys Ser Asp Ser Val Glu Tyr Ile Ile Ile Thr Asp Ser Glu Gly Ile 
145                 150                 155                 160 


Leu Gly Ile Gly Asp Gln Gly Val Gly Gly Val Leu Ile Ser Val Ala 
                165                 170                 175     


Lys Gly His Leu Met Thr Leu Cys Ala Gly Leu Asp Pro Asn Arg Phe 
            180                 185                 190         


Leu Pro Ile Val Leu Asp Val Gly Thr Asn Asn Glu Thr His Arg Lys 
        195                 200                 205             


Asn His Gln Tyr Met Gly Leu Arg Lys Asp Arg Val Arg Gly Glu Gln 
    210                 215                 220                 


Tyr Asp Ser Phe Leu Asp Asn Val Ile Lys Ala Ile Arg Glu Val Phe 
225                 230                 235                 240 


Pro Glu Ala Phe Ile His Phe Glu Asp Phe Gly Leu Ala Asn Ala Lys 
                245                 250                 255     


Arg Ile Leu Asp His Tyr Arg Pro Asp Ile Ala Cys Phe Asn Asp Asp 
            260                 265                 270         


Ile Gln Gly Thr Gly Ala Val Ala Leu Ala Ala Ile Ile Gly Ala Leu 
        275                 280                 285             


His Val Thr Lys Ser Pro Leu Thr Glu Gln Arg Ile Met Ile Phe Gly 
    290                 295                 300                 


Ala Gly Thr Ala Gly Val Gly Ile Ala Asn Gln Ile Val Ala Gly Met 
305                 310                 315                 320 


Val Thr Asp Gly Leu Ser Leu Asp Lys Ala Arg Gly Asn Leu Phe Met 
                325                 330                 335     


Ile Asp Arg Cys Gly Leu Leu Leu Glu Arg His Ala Lys Ile Ala Thr 
            340                 345                 350         


Asp Gly Gln Lys Pro Phe Leu Lys Lys Asp Ser Asp Phe Lys Glu Val 
        355                 360                 365             


Pro Ser Gly Asp Ile Asn Leu Glu Ser Ala Ile Ala Leu Val Lys Pro 
    370                 375                 380                 


Thr Ile Leu Leu Gly Cys Ser Gly Gln Pro Gly Lys Phe Thr Glu Lys 
385                 390                 395                 400 


Ala Ile Arg Glu Met Ser Lys His Val Glu Arg Pro Ile Ile Phe Pro 
                405                 410                 415     


Ile Ser Asn Pro Thr Thr Leu Met Glu Ala Lys Pro Asp Gln Ile Asp 
            420                 425                 430         


Lys Trp Ser Asp Gly Lys Ala Leu Ile Ala Thr Gly Ser Pro Leu Pro 
        435                 440                 445             


Pro Leu Asn Arg Asn Gly Lys Lys Tyr Val Ile Ser Gln Cys Asn Asn 
    450                 455                 460                 


Ala Leu Leu Tyr Pro Ala Leu Gly Val Ala Cys Val Leu Ser Arg Cys 
465                 470                 475                 480 


Lys Leu Leu Ser Asp Gly Met Leu Lys Ala Ala Ser Asp Ala Leu Ala 
                485                 490                 495     


Thr Val Pro Arg Ser Leu Phe Ala Ala Asp Glu Ala Leu Leu Pro Asp 
            500                 505                 510         


Leu Asn Asn Ala Arg Glu Ile Ser Arg His Ile Val Phe Ala Val Leu 
        515                 520                 525             


Lys Gln Ala Val Ser Glu Gly Met Ser Thr Val Asp Leu Pro Lys Asp 
    530                 535                 540                 


Asp Ala Lys Leu Lys Glu Trp Ile Ile Glu Arg Glu Trp Asn Pro Glu 
545                 550                 555                 560 


Tyr Lys Pro Phe Val 
                565 


<210>  4
<211>  1737
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Cytosolic malic enzyme, YlME-T2 (used in pME-T2 construct)


<220>
<221>  CDS
<222>  (1)..(1737)

<400>  4
atg gct aca acc cgt ctg tcg gtt gag ggc ccc atc tac gtg ggc ttc         48
Met Ala Thr Thr Arg Leu Ser Val Glu Gly Pro Ile Tyr Val Gly Phe           
1               5                   10                  15                

gac ggc att cgt ctt ctc aac ctg ccg cat ctc aac aag ggc tcg gga         96
Asp Gly Ile Arg Leu Leu Asn Leu Pro His Leu Asn Lys Gly Ser Gly           
            20                  25                  30                    

ttc ccc ctc aac gag cga cgg gaa ttc aga ctc agt ggt ctt ctg ccc        144
Phe Pro Leu Asn Glu Arg Arg Glu Phe Arg Leu Ser Gly Leu Leu Pro           
        35                  40                  45                        

tct gcc gaa gcc acc ctg gag gaa cag gtc gac cga gca tac caa caa        192
Ser Ala Glu Ala Thr Leu Glu Glu Gln Val Asp Arg Ala Tyr Gln Gln           
    50                  55                  60                            

ttc aaa aag tgt ggc act ccc tta gcc aaa aac ggg ttc tgc acc tcg        240
Phe Lys Lys Cys Gly Thr Pro Leu Ala Lys Asn Gly Phe Cys Thr Ser           
65                  70                  75                  80            

ctc aag ttc caa aac gag gtg ctc tac tac gcc ctg ctg ctc aag cac        288
Leu Lys Phe Gln Asn Glu Val Leu Tyr Tyr Ala Leu Leu Leu Lys His           
                85                  90                  95                

gtt aag gag gtc ttc ccc atc atc tat aca ccg act cag gga gaa gcc        336
Val Lys Glu Val Phe Pro Ile Ile Tyr Thr Pro Thr Gln Gly Glu Ala           
            100                 105                 110                   

att gaa cag tac tcg cgg ctg ttc cgg cgg ccc gaa ggc tgc ttc ctc        384
Ile Glu Gln Tyr Ser Arg Leu Phe Arg Arg Pro Glu Gly Cys Phe Leu           
        115                 120                 125                       

gac atc acc agt ccc tac gac gtg gag gag cgt ctg gga gcg ttt gga        432
Asp Ile Thr Ser Pro Tyr Asp Val Glu Glu Arg Leu Gly Ala Phe Gly           
    130                 135                 140                           

gac cat gac gac att gac tac att gtc gtg act gac tcc gag ggt att        480
Asp His Asp Asp Ile Asp Tyr Ile Val Val Thr Asp Ser Glu Gly Ile           
145                 150                 155                 160           

ctc gga att gga gac caa gga gtg ggc ggt att ggt att tcc atc gcc        528
Leu Gly Ile Gly Asp Gln Gly Val Gly Gly Ile Gly Ile Ser Ile Ala           
                165                 170                 175               

aag ctg gct ctc atg act cta tgt gct gga gtc aac ccc tca cga gtc        576
Lys Leu Ala Leu Met Thr Leu Cys Ala Gly Val Asn Pro Ser Arg Val           
            180                 185                 190                   

att cct gtg gtt ctg gat acg gga acc aac aac cag gag ctg ctg cac        624
Ile Pro Val Val Leu Asp Thr Gly Thr Asn Asn Gln Glu Leu Leu His           
        195                 200                 205                       

gac ccc ctg tat ctc ggc cga cga atg ccc cga gtg cga gga aag cag        672
Asp Pro Leu Tyr Leu Gly Arg Arg Met Pro Arg Val Arg Gly Lys Gln           
    210                 215                 220                           

tac gac gac ttc atc gac aac ttt gtg cag tct gcc cga agg ctg tat        720
Tyr Asp Asp Phe Ile Asp Asn Phe Val Gln Ser Ala Arg Arg Leu Tyr           
225                 230                 235                 240           

ccc aag gcg gtg atc cat ttc gag gac ttt ggg ctc gct aac gca cac        768
Pro Lys Ala Val Ile His Phe Glu Asp Phe Gly Leu Ala Asn Ala His           
                245                 250                 255               

aag atc ctc gac aag tat cga ccg gag atc ccc tgc ttc aac gac gac        816
Lys Ile Leu Asp Lys Tyr Arg Pro Glu Ile Pro Cys Phe Asn Asp Asp           
            260                 265                 270                   

atc cag ggc act gga gcc gtc act ttg gcc tcc atc acg gcc gct ctc        864
Ile Gln Gly Thr Gly Ala Val Thr Leu Ala Ser Ile Thr Ala Ala Leu           
        275                 280                 285                       

aag gtg ctg ggc aaa aat atc aca gat act cga att ctc gtg tac gga        912
Lys Val Leu Gly Lys Asn Ile Thr Asp Thr Arg Ile Leu Val Tyr Gly           
    290                 295                 300                           

gct ggt tcg gcc ggc atg ggt att gct gaa cag gtc tat gat aac ctg        960
Ala Gly Ser Ala Gly Met Gly Ile Ala Glu Gln Val Tyr Asp Asn Leu           
305                 310                 315                 320           

gtt gcc cag ggt ctc gac gac aag act gcg cga caa aac atc ttt ctc       1008
Val Ala Gln Gly Leu Asp Asp Lys Thr Ala Arg Gln Asn Ile Phe Leu           
                325                 330                 335               

atg gac cga ccg ggt cta ctg acc acc gca ctt acc gac gag cag atg       1056
Met Asp Arg Pro Gly Leu Leu Thr Thr Ala Leu Thr Asp Glu Gln Met           
            340                 345                 350                   

agc gac gtg cag aag ccg ttt gcc aag gac aag gcc aat tac gag gga       1104
Ser Asp Val Gln Lys Pro Phe Ala Lys Asp Lys Ala Asn Tyr Glu Gly           
        355                 360                 365                       

gtg gac acc aag act ctg gag cac gtg gtt gct gcc gtc aag ccc cat       1152
Val Asp Thr Lys Thr Leu Glu His Val Val Ala Ala Val Lys Pro His           
    370                 375                 380                           

att ctc att gga tgt tcc act cag ccc ggc gcc ttt aac gag aag gtc       1200
Ile Leu Ile Gly Cys Ser Thr Gln Pro Gly Ala Phe Asn Glu Lys Val           
385                 390                 395                 400           

gtc aag gag atg ctc aaa cac acc cct cga ccc atc att ctc cct ctt       1248
Val Lys Glu Met Leu Lys His Thr Pro Arg Pro Ile Ile Leu Pro Leu           
                405                 410                 415               

tcc aac ccc aca cgt ctt cat gag gct gtc cct gca gat ctg tac aag       1296
Ser Asn Pro Thr Arg Leu His Glu Ala Val Pro Ala Asp Leu Tyr Lys           
            420                 425                 430                   

tgg acc gac ggc aag gct ctg gtt gcc acc ggc tcg ccc ttt gac cca       1344
Trp Thr Asp Gly Lys Ala Leu Val Ala Thr Gly Ser Pro Phe Asp Pro           
        435                 440                 445                       

gtc aac ggc aag gag acg tct gag aac aat aac tgc ttt gtt ttc ccc       1392
Val Asn Gly Lys Glu Thr Ser Glu Asn Asn Asn Cys Phe Val Phe Pro           
    450                 455                 460                           

gga atc ggg ctg gga gcc att ctg tct cga tca aag ctc atc acc aac       1440
Gly Ile Gly Leu Gly Ala Ile Leu Ser Arg Ser Lys Leu Ile Thr Asn           
465                 470                 475                 480           

acc atg att gct gct gcc atc gag tgc ctc gcc gaa cag gcc ccc att       1488
Thr Met Ile Ala Ala Ala Ile Glu Cys Leu Ala Glu Gln Ala Pro Ile           
                485                 490                 495               

ctc aag aac cac gac gag gga gta ctt ccc gac gta gct ctc atc cag       1536
Leu Lys Asn His Asp Glu Gly Val Leu Pro Asp Val Ala Leu Ile Gln           
            500                 505                 510                   

atc att tcg gcc cgg gtg gcc act gcc gtg gtt ctt cag gcc aag gct       1584
Ile Ile Ser Ala Arg Val Ala Thr Ala Val Val Leu Gln Ala Lys Ala           
        515                 520                 525                       

gag ggc cta gcc act gtc gag gaa gag ctc aag ccc ggc acc aag gaa       1632
Glu Gly Leu Ala Thr Val Glu Glu Glu Leu Lys Pro Gly Thr Lys Glu           
    530                 535                 540                           

cat gtg cag att ccc gac aac ttt gac gag tgt ctc gcc tgg gtc gag       1680
His Val Gln Ile Pro Asp Asn Phe Asp Glu Cys Leu Ala Trp Val Glu           
545                 550                 555                 560           

act cag atg tgg cgg ccc gtc tac cgg cct ctc atc cat gtg cgg gat       1728
Thr Gln Met Trp Arg Pro Val Tyr Arg Pro Leu Ile His Val Arg Asp           
                565                 570                 575               

tac gac tag                                                           1737
Tyr Asp                                                                   
                                                                          


<210>  5
<211>  578
<212>  PRT
<213>  Artificial sequence

<220>
<223>  Synthetic Construct

<400>  5

Met Ala Thr Thr Arg Leu Ser Val Glu Gly Pro Ile Tyr Val Gly Phe 
1               5                   10                  15      


Asp Gly Ile Arg Leu Leu Asn Leu Pro His Leu Asn Lys Gly Ser Gly 
            20                  25                  30          


Phe Pro Leu Asn Glu Arg Arg Glu Phe Arg Leu Ser Gly Leu Leu Pro 
        35                  40                  45              


Ser Ala Glu Ala Thr Leu Glu Glu Gln Val Asp Arg Ala Tyr Gln Gln 
    50                  55                  60                  


Phe Lys Lys Cys Gly Thr Pro Leu Ala Lys Asn Gly Phe Cys Thr Ser 
65                  70                  75                  80  


Leu Lys Phe Gln Asn Glu Val Leu Tyr Tyr Ala Leu Leu Leu Lys His 
                85                  90                  95      


Val Lys Glu Val Phe Pro Ile Ile Tyr Thr Pro Thr Gln Gly Glu Ala 
            100                 105                 110         


Ile Glu Gln Tyr Ser Arg Leu Phe Arg Arg Pro Glu Gly Cys Phe Leu 
        115                 120                 125             


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


Asp His Asp Asp Ile Asp Tyr Ile Val Val Thr Asp Ser Glu Gly Ile 
145                 150                 155                 160 


Leu Gly Ile Gly Asp Gln Gly Val Gly Gly Ile Gly Ile Ser Ile Ala 
                165                 170                 175     


Lys Leu Ala Leu Met Thr Leu Cys Ala Gly Val Asn Pro Ser Arg Val 
            180                 185                 190         


Ile Pro Val Val Leu Asp Thr Gly Thr Asn Asn Gln Glu Leu Leu His 
        195                 200                 205             


Asp Pro Leu Tyr Leu Gly Arg Arg Met Pro Arg Val Arg Gly Lys Gln 
    210                 215                 220                 


Tyr Asp Asp Phe Ile Asp Asn Phe Val Gln Ser Ala Arg Arg Leu Tyr 
225                 230                 235                 240 


Pro Lys Ala Val Ile His Phe Glu Asp Phe Gly Leu Ala Asn Ala His 
                245                 250                 255     


Lys Ile Leu Asp Lys Tyr Arg Pro Glu Ile Pro Cys Phe Asn Asp Asp 
            260                 265                 270         


Ile Gln Gly Thr Gly Ala Val Thr Leu Ala Ser Ile Thr Ala Ala Leu 
        275                 280                 285             


Lys Val Leu Gly Lys Asn Ile Thr Asp Thr Arg Ile Leu Val Tyr Gly 
    290                 295                 300                 


Ala Gly Ser Ala Gly Met Gly Ile Ala Glu Gln Val Tyr Asp Asn Leu 
305                 310                 315                 320 


Val Ala Gln Gly Leu Asp Asp Lys Thr Ala Arg Gln Asn Ile Phe Leu 
                325                 330                 335     


Met Asp Arg Pro Gly Leu Leu Thr Thr Ala Leu Thr Asp Glu Gln Met 
            340                 345                 350         


Ser Asp Val Gln Lys Pro Phe Ala Lys Asp Lys Ala Asn Tyr Glu Gly 
        355                 360                 365             


Val Asp Thr Lys Thr Leu Glu His Val Val Ala Ala Val Lys Pro His 
    370                 375                 380                 


Ile Leu Ile Gly Cys Ser Thr Gln Pro Gly Ala Phe Asn Glu Lys Val 
385                 390                 395                 400 


Val Lys Glu Met Leu Lys His Thr Pro Arg Pro Ile Ile Leu Pro Leu 
                405                 410                 415     


Ser Asn Pro Thr Arg Leu His Glu Ala Val Pro Ala Asp Leu Tyr Lys 
            420                 425                 430         


Trp Thr Asp Gly Lys Ala Leu Val Ala Thr Gly Ser Pro Phe Asp Pro 
        435                 440                 445             


Val Asn Gly Lys Glu Thr Ser Glu Asn Asn Asn Cys Phe Val Phe Pro 
    450                 455                 460                 


Gly Ile Gly Leu Gly Ala Ile Leu Ser Arg Ser Lys Leu Ile Thr Asn 
465                 470                 475                 480 


Thr Met Ile Ala Ala Ala Ile Glu Cys Leu Ala Glu Gln Ala Pro Ile 
                485                 490                 495     


Leu Lys Asn His Asp Glu Gly Val Leu Pro Asp Val Ala Leu Ile Gln 
            500                 505                 510         


Ile Ile Ser Ala Arg Val Ala Thr Ala Val Val Leu Gln Ala Lys Ala 
        515                 520                 525             


Glu Gly Leu Ala Thr Val Glu Glu Glu Leu Lys Pro Gly Thr Lys Glu 
    530                 535                 540                 


His Val Gln Ile Pro Asp Asn Phe Asp Glu Cys Leu Ala Trp Val Glu 
545                 550                 555                 560 


Thr Gln Met Trp Arg Pro Val Tyr Arg Pro Leu Ile His Val Arg Asp 
                565                 570                 575     


Tyr Asp 
        


<210>  6
<211>  1890
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Malic enzyme, YlME (used in pME construct)


<220>
<221>  CDS
<222>  (1)..(1890)

<400>  6
atg gta cga cta cga acc atg cga ccc aca cag acc agc gtc agg gcg         48
Met Val Arg Leu Arg Thr Met Arg Pro Thr Gln Thr Ser Val Arg Ala           
1               5                   10                  15                

gcg ctt ggg ccc acc gcc gcg gcc cga aac atg tcc tcc tcc agc ccc         96
Ala Leu Gly Pro Thr Ala Ala Ala Arg Asn Met Ser Ser Ser Ser Pro           
            20                  25                  30                    

tcc agc ttc gaa tac tcg tcc tac gtc aag ggc acg cgg gaa atc ggc        144
Ser Ser Phe Glu Tyr Ser Ser Tyr Val Lys Gly Thr Arg Glu Ile Gly           
        35                  40                  45                        

cac cga aag gcg ccc aca acc cgt ctg tcg gtt gag ggc ccc atc tac        192
His Arg Lys Ala Pro Thr Thr Arg Leu Ser Val Glu Gly Pro Ile Tyr           
    50                  55                  60                            

gtg ggc ttc gac ggc att cgt ctt ctc aac ctg ccg cat ctc aac aag        240
Val Gly Phe Asp Gly Ile Arg Leu Leu Asn Leu Pro His Leu Asn Lys           
65                  70                  75                  80            

ggc tcg gga ttc ccc ctc aac gag cga cgg gaa ttc aga ctc agt ggt        288
Gly Ser Gly Phe Pro Leu Asn Glu Arg Arg Glu Phe Arg Leu Ser Gly           
                85                  90                  95                

ctt ctg ccc tct gcc gaa gcc acc ctg gag gaa cag gtc gac cga gca        336
Leu Leu Pro Ser Ala Glu Ala Thr Leu Glu Glu Gln Val Asp Arg Ala           
            100                 105                 110                   

tac caa caa ttc aaa aag tgt ggc act ccc tta gcc aaa aac ggg ttc        384
Tyr Gln Gln Phe Lys Lys Cys Gly Thr Pro Leu Ala Lys Asn Gly Phe           
        115                 120                 125                       

tgc acc tcg ctc aag ttc caa aac gag gtg ctc tac tac gcc ctg ctg        432
Cys Thr Ser Leu Lys Phe Gln Asn Glu Val Leu Tyr Tyr Ala Leu Leu           
    130                 135                 140                           

ctc aag cac gtt aag gag gtc ttc ccc atc atc tat aca ccg act cag        480
Leu Lys His Val Lys Glu Val Phe Pro Ile Ile Tyr Thr Pro Thr Gln           
145                 150                 155                 160           

gga gaa gcc att gaa cag tac tcg cgg ctg ttc cgg cgg ccc gaa ggc        528
Gly Glu Ala Ile Glu Gln Tyr Ser Arg Leu Phe Arg Arg Pro Glu Gly           
                165                 170                 175               

tgc ttc ctc gac atc acc agt ccc tac gac gtg gag gag cgt ctg gga        576
Cys Phe Leu Asp Ile Thr Ser Pro Tyr Asp Val Glu Glu Arg Leu Gly           
            180                 185                 190                   

gcg ttt gga gac cat gac gac att gac tac att gtc gtg act gac tcc        624
Ala Phe Gly Asp His Asp Asp Ile Asp Tyr Ile Val Val Thr Asp Ser           
        195                 200                 205                       

gag ggt att ctc gga att gga gac caa gga gtg ggc ggt att ggt att        672
Glu Gly Ile Leu Gly Ile Gly Asp Gln Gly Val Gly Gly Ile Gly Ile           
    210                 215                 220                           

tcc atc gcc aag ctg gct ctc atg act cta tgt gct gga gtc aac ccc        720
Ser Ile Ala Lys Leu Ala Leu Met Thr Leu Cys Ala Gly Val Asn Pro           
225                 230                 235                 240           

tca cga gtc att cct gtg gtt ctg gat acg gga acc aac aac cag gag        768
Ser Arg Val Ile Pro Val Val Leu Asp Thr Gly Thr Asn Asn Gln Glu           
                245                 250                 255               

ctg ctg cac gac ccc ctg tat ctc ggc cga cga atg ccc cga gtg cga        816
Leu Leu His Asp Pro Leu Tyr Leu Gly Arg Arg Met Pro Arg Val Arg           
            260                 265                 270                   

gga aag cag tac gac gac ttc atc gac aac ttt gtg cag tct gcc cga        864
Gly Lys Gln Tyr Asp Asp Phe Ile Asp Asn Phe Val Gln Ser Ala Arg           
        275                 280                 285                       

agg ctg tat ccc aag gcg gtg atc cat ttc gag gac ttt ggg ctc gct        912
Arg Leu Tyr Pro Lys Ala Val Ile His Phe Glu Asp Phe Gly Leu Ala           
    290                 295                 300                           

aac gca cac aag atc ctc gac aag tat cga ccg gag atc ccc tgc ttc        960
Asn Ala His Lys Ile Leu Asp Lys Tyr Arg Pro Glu Ile Pro Cys Phe           
305                 310                 315                 320           

aac gac gac atc cag ggc act gga gcc gtc act ttg gcc tcc atc acg       1008
Asn Asp Asp Ile Gln Gly Thr Gly Ala Val Thr Leu Ala Ser Ile Thr           
                325                 330                 335               

gcc gct ctc aag gtg ctg ggc aaa aat atc aca gat act cga att ctc       1056
Ala Ala Leu Lys Val Leu Gly Lys Asn Ile Thr Asp Thr Arg Ile Leu           
            340                 345                 350                   

gtg tac gga gct ggt tcg gcc ggc atg ggt att gct gaa cag gtc tat       1104
Val Tyr Gly Ala Gly Ser Ala Gly Met Gly Ile Ala Glu Gln Val Tyr           
        355                 360                 365                       

gat aac ctg gtt gcc cag ggt ctc gac gac aag act gcg cga caa aac       1152
Asp Asn Leu Val Ala Gln Gly Leu Asp Asp Lys Thr Ala Arg Gln Asn           
    370                 375                 380                           

atc ttt ctc atg gac cga ccg ggt cta ctg acc acc gca ctt acc gac       1200
Ile Phe Leu Met Asp Arg Pro Gly Leu Leu Thr Thr Ala Leu Thr Asp           
385                 390                 395                 400           

gag cag atg agc gac gtg cag aag ccg ttt gcc aag gac aag gcc aat       1248
Glu Gln Met Ser Asp Val Gln Lys Pro Phe Ala Lys Asp Lys Ala Asn           
                405                 410                 415               

tac gag gga gtg gac acc aag act ctg gag cac gtg gtt gct gcc gtc       1296
Tyr Glu Gly Val Asp Thr Lys Thr Leu Glu His Val Val Ala Ala Val           
            420                 425                 430                   

aag ccc cat att ctc att gga tgt tcc act cag ccc ggc gcc ttt aac       1344
Lys Pro His Ile Leu Ile Gly Cys Ser Thr Gln Pro Gly Ala Phe Asn           
        435                 440                 445                       

gag aag gtc gtc aag gag atg ctc aaa cac acc cct cga ccc atc att       1392
Glu Lys Val Val Lys Glu Met Leu Lys His Thr Pro Arg Pro Ile Ile           
    450                 455                 460                           

ctc cct ctt tcc aac ccc aca cgt ctt cat gag gct gtc cct gca gat       1440
Leu Pro Leu Ser Asn Pro Thr Arg Leu His Glu Ala Val Pro Ala Asp           
465                 470                 475                 480           

ctg tac aag tgg acc gac ggc aag gct ctg gtt gcc acc ggc tcg ccc       1488
Leu Tyr Lys Trp Thr Asp Gly Lys Ala Leu Val Ala Thr Gly Ser Pro           
                485                 490                 495               

ttt gac cca gtc aac ggc aag gag acg tct gag aac aat aac tgc ttt       1536
Phe Asp Pro Val Asn Gly Lys Glu Thr Ser Glu Asn Asn Asn Cys Phe           
            500                 505                 510                   

gtt ttc ccc gga atc ggg ctg gga gcc att ctg tct cga tca aag ctc       1584
Val Phe Pro Gly Ile Gly Leu Gly Ala Ile Leu Ser Arg Ser Lys Leu           
        515                 520                 525                       

atc acc aac acc atg att gct gct gcc atc gag tgc ctc gcc gaa cag       1632
Ile Thr Asn Thr Met Ile Ala Ala Ala Ile Glu Cys Leu Ala Glu Gln           
    530                 535                 540                           

gcc ccc att ctc aag aac cac gac gag gga gta ctt ccc gac gta gct       1680
Ala Pro Ile Leu Lys Asn His Asp Glu Gly Val Leu Pro Asp Val Ala           
545                 550                 555                 560           

ctc atc cag atc att tcg gcc cgg gtg gcc act gcc gtg gtt ctt cag       1728
Leu Ile Gln Ile Ile Ser Ala Arg Val Ala Thr Ala Val Val Leu Gln           
                565                 570                 575               

gcc aag gct gag ggc cta gcc act gtc gag gaa gag ctc aag ccc ggc       1776
Ala Lys Ala Glu Gly Leu Ala Thr Val Glu Glu Glu Leu Lys Pro Gly           
            580                 585                 590                   

acc aag gaa cat gtg cag att ccc gac aac ttt gac gag tgt ctc gcc       1824
Thr Lys Glu His Val Gln Ile Pro Asp Asn Phe Asp Glu Cys Leu Ala           
        595                 600                 605                       

tgg gtc gag act cag atg tgg cgg ccc gtc tac cgg cct ctc atc cat       1872
Trp Val Glu Thr Gln Met Trp Arg Pro Val Tyr Arg Pro Leu Ile His           
    610                 615                 620                           

gtg cgg gat tac gac tag                                               1890
Val Arg Asp Tyr Asp                                                       
625                                                                       


<210>  7
<211>  629
<212>  PRT
<213>  Artificial sequence

<220>
<223>  Synthetic Construct

<400>  7

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


Ala Leu Gly Pro Thr Ala Ala Ala Arg Asn Met Ser Ser Ser Ser Pro 
            20                  25                  30          


Ser Ser Phe Glu Tyr Ser Ser Tyr Val Lys Gly Thr Arg Glu Ile Gly 
        35                  40                  45              


His Arg Lys Ala Pro Thr Thr Arg Leu Ser Val Glu Gly Pro Ile Tyr 
    50                  55                  60                  


Val Gly Phe Asp Gly Ile Arg Leu Leu Asn Leu Pro His Leu Asn Lys 
65                  70                  75                  80  


Gly Ser Gly Phe Pro Leu Asn Glu Arg Arg Glu Phe Arg Leu Ser Gly 
                85                  90                  95      


Leu Leu Pro Ser Ala Glu Ala Thr Leu Glu Glu Gln Val Asp Arg Ala 
            100                 105                 110         


Tyr Gln Gln Phe Lys Lys Cys Gly Thr Pro Leu Ala Lys Asn Gly Phe 
        115                 120                 125             


Cys Thr Ser Leu Lys Phe Gln Asn Glu Val Leu Tyr Tyr Ala Leu Leu 
    130                 135                 140                 


Leu Lys His Val Lys Glu Val Phe Pro Ile Ile Tyr Thr Pro Thr Gln 
145                 150                 155                 160 


Gly Glu Ala Ile Glu Gln Tyr Ser Arg Leu Phe Arg Arg Pro Glu Gly 
                165                 170                 175     


Cys Phe Leu Asp Ile Thr Ser Pro Tyr Asp Val Glu Glu Arg Leu Gly 
            180                 185                 190         


Ala Phe Gly Asp His Asp Asp Ile Asp Tyr Ile Val Val Thr Asp Ser 
        195                 200                 205             


Glu Gly Ile Leu Gly Ile Gly Asp Gln Gly Val Gly Gly Ile Gly Ile 
    210                 215                 220                 


Ser Ile Ala Lys Leu Ala Leu Met Thr Leu Cys Ala Gly Val Asn Pro 
225                 230                 235                 240 


Ser Arg Val Ile Pro Val Val Leu Asp Thr Gly Thr Asn Asn Gln Glu 
                245                 250                 255     


Leu Leu His Asp Pro Leu Tyr Leu Gly Arg Arg Met Pro Arg Val Arg 
            260                 265                 270         


Gly Lys Gln Tyr Asp Asp Phe Ile Asp Asn Phe Val Gln Ser Ala Arg 
        275                 280                 285             


Arg Leu Tyr Pro Lys Ala Val Ile His Phe Glu Asp Phe Gly Leu Ala 
    290                 295                 300                 


Asn Ala His Lys Ile Leu Asp Lys Tyr Arg Pro Glu Ile Pro Cys Phe 
305                 310                 315                 320 


Asn Asp Asp Ile Gln Gly Thr Gly Ala Val Thr Leu Ala Ser Ile Thr 
                325                 330                 335     


Ala Ala Leu Lys Val Leu Gly Lys Asn Ile Thr Asp Thr Arg Ile Leu 
            340                 345                 350         


Val Tyr Gly Ala Gly Ser Ala Gly Met Gly Ile Ala Glu Gln Val Tyr 
        355                 360                 365             


Asp Asn Leu Val Ala Gln Gly Leu Asp Asp Lys Thr Ala Arg Gln Asn 
    370                 375                 380                 


Ile Phe Leu Met Asp Arg Pro Gly Leu Leu Thr Thr Ala Leu Thr Asp 
385                 390                 395                 400 


Glu Gln Met Ser Asp Val Gln Lys Pro Phe Ala Lys Asp Lys Ala Asn 
                405                 410                 415     


Tyr Glu Gly Val Asp Thr Lys Thr Leu Glu His Val Val Ala Ala Val 
            420                 425                 430         


Lys Pro His Ile Leu Ile Gly Cys Ser Thr Gln Pro Gly Ala Phe Asn 
        435                 440                 445             


Glu Lys Val Val Lys Glu Met Leu Lys His Thr Pro Arg Pro Ile Ile 
    450                 455                 460                 


Leu Pro Leu Ser Asn Pro Thr Arg Leu His Glu Ala Val Pro Ala Asp 
465                 470                 475                 480 


Leu Tyr Lys Trp Thr Asp Gly Lys Ala Leu Val Ala Thr Gly Ser Pro 
                485                 490                 495     


Phe Asp Pro Val Asn Gly Lys Glu Thr Ser Glu Asn Asn Asn Cys Phe 
            500                 505                 510         


Val Phe Pro Gly Ile Gly Leu Gly Ala Ile Leu Ser Arg Ser Lys Leu 
        515                 520                 525             


Ile Thr Asn Thr Met Ile Ala Ala Ala Ile Glu Cys Leu Ala Glu Gln 
    530                 535                 540                 


Ala Pro Ile Leu Lys Asn His Asp Glu Gly Val Leu Pro Asp Val Ala 
545                 550                 555                 560 


Leu Ile Gln Ile Ile Ser Ala Arg Val Ala Thr Ala Val Val Leu Gln 
                565                 570                 575     


Ala Lys Ala Glu Gly Leu Ala Thr Val Glu Glu Glu Leu Lys Pro Gly 
            580                 585                 590         


Thr Lys Glu His Val Gln Ile Pro Asp Asn Phe Asp Glu Cys Leu Ala 
        595                 600                 605             


Trp Val Glu Thr Gln Met Trp Arg Pro Val Tyr Arg Pro Leu Ile His 
    610                 615                 620                 


Val Arg Asp Tyr Asp 
625                 


<210>  8
<211>  33
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Primer ME-TN2

<400>  8
gatcccatgg ctacaacccg tctgtcggtt gag                                    33


<210>  9
<211>  33
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Primer ME-T2

<400>  9
gatcgcggcc gctgcttact aaactaaact gtc                                    33


<210>  10
<211>  8986
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Plasmid construct pMET2

<400>  10
ggccgcaagt gtggatgggg aagtgagtgc ccggttctgt gtgcacaatt ggcaatccaa       60

gatggatgga ttcaacacag ggatatagcg agctacgtgg tggtgcgagg atatagcaac      120

ggatatttat gtttgacact tgagaatgta cgatacaagc actgtccaag tacaatacta      180

aacatactgt acatactcat actcgtaccc gggcaacggt ttcacttgag tgcagtggct      240

agtgctctta ctcgtacagt gtgcaatact gcgtatcata gtctttgatg tatatcgtat      300

tcattcatgt tagttgcgta cgggcgtcgt tgcttgtgtg atttttgagg acccatccct      360

ttggtatata agtatactct ggggttaagg ttgcccgtgt agtctaggtt atagttttca      420

tgtgaaatac cgagagccga gggagaataa acgggggtat ttggacttgt ttttttcgcg      480

gaaaagcgtc gaatcaaccc tgcgggcctt gcaccatgtc cacgacgtgt ttctcgcccc      540

aattcgcccc ttgcacgtca aaattaggcc tccatctaga cccctccata acatgtgact      600

gtggggaaaa gtataaggga aaccatgcaa ccatagacga cgtgaaagac ggggaggaac      660

caatggaggc caaagaaatg gggtagcaac agtccaggag acagacaagg agacaaggag      720

agggcgcccg aaagatcgga aaaacaaaca tgtccaattg gggcagtgac ggaaacgaca      780

cggacacttc agtacaatgg accgaccatc tccaagccag ggttattccg gtatcacctt      840

ggccgtaacc tcccgctggt acctgatatt gtacacgttc acattcaata tactttcagc      900

tacaataaga gaggctgttt gtcgggcatg tgtgtccgtc gtatggggtg atgtccgagg      960

gcgaaattcg ctacaagctt aactctggcg cttgtccagt atgaatagac aagtcaagac     1020

cagtggtgcc atgattgaca gggaggtaca agacttcgat actcgagcat tactcggact     1080

tgtggcgatt gaacagacgg gcgatcgctt ctcccccgta ttgccggcgc gccagctgca     1140

ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc     1200

ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc     1260

aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc     1320

aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag     1380

gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc     1440

gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt     1500

tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct     1560

ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg     1620

ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct     1680

tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat     1740

tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg     1800

ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa     1860

aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt     1920

ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc     1980

tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt     2040

atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta     2100

aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat     2160

ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac     2220

tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg     2280

ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag     2340

tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt     2400

aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt     2460

gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt     2520

tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt     2580

cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct     2640

tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt     2700

ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac     2760

cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa     2820

actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa     2880

ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca     2940

aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct     3000

ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga     3060

atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc     3120

tgatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggaaattgt     3180

aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa     3240

ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt     3300

gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa     3360

agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag     3420

ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt     3480

tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg     3540

agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc     3600

cgcgcttaat gcgccgctac agggcgcgtc cattcgccat tcaggctgcg caactgttgg     3660

gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct     3720

gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg     3780

gccagtgaat tgtaatacga ctcactatag ggcgaattgg gcccgacgtc gcatgcgctg     3840

atgacacttt ggtctgaaag agatgcattt tgaatcccaa acttgcagtg cccaagtgac     3900

atacatctcc gcgttttgga aaatgttcag aaacagttga ttgtgttgga atggggaatg     3960

gggaatggaa aaatgactca agtatcaatt ccaaaaactt ctctggctgg cagtacctac     4020

tgtccatact actgcatttt ctccagtcag gccactctat actcgacgac acagtagtaa     4080

aacccagata atttcgacat aaacaagaaa acagacccaa taatatttat atatagtcag     4140

ccgtttgtcc agttcagact gtaatagccg aaaaaaaatc caaagtttct attctaggaa     4200

aatatattcc aatattttta attcttaatc tcatttattt tattctagcg aaatacattt     4260

cagctacttg agacatgtga tacccacaaa tcggattcgg actcggttgt tcagaagagc     4320

atatggcatt cgtgctcgct tgttcacgta ttcttcctgt tccatctctt ggccgacaat     4380

cacacaaaaa tggggttttt tttttaattc taatgattca ttacagcaaa attgagatat     4440

agcagaccac gtattccata atcaccaagg aagttcttgg gcgtcttaat taagtcatac     4500

acaagtcagc tttcttcgag cctcatataa gtataagtag ttcaacgtat tagcactgta     4560

cccagcatct ccgtatcgag aaacacaaca acatgcccca ttggacagat catgcggata     4620

cacaggttgt gcagtatcat acatactcga tcagacaggt cgtctgacca tcatacaagc     4680

tgaacaagcg ctccatactt gcacgctctc tatatacaca gttaaattac atatccatag     4740

tctaacctct aacagttaat cttctggtaa gcctcccagc cagccttctg gtatcgcttg     4800

gcctcctcaa taggatctcg gttctggccg tacagacctc ggccgacaat tatgatatcc     4860

gttccggtag acatgacatc ctcaacagtt cggtactgct gtccgagagc gtctcccttg     4920

tcgtcaagac ccaccccggg ggtcagaata agccagtcct cagagtcgcc cttaggtcgg     4980

ttctgggcaa tgaagccaac cacaaactcg gggtcggatc gggcaagctc aatggtctgc     5040

ttggagtact cgccagtggc cagagagccc ttgcaagaca gctcggccag catgagcaga     5100

cctctggcca gcttctcgtt gggagagggg actaggaact ccttgtactg ggagttctcg     5160

tagtcagaga cgtcctcctt cttctgttca gagacagttt cctcggcacc agctcgcagg     5220

ccagcaatga ttccggttcc gggtacaccg tgggcgttgg tgatatcgga ccactcggcg     5280

attcggtgac accggtactg gtgcttgaca gtgttgccaa tatctgcgaa ctttctgtcc     5340

tcgaacagga agaaaccgtg cttaagagca agttccttga gggggagcac agtgccggcg     5400

taggtgaagt cgtcaatgat gtcgatatgg gttttgatca tgcacacata aggtccgacc     5460

ttatcggcaa gctcaatgag ctccttggtg gtggtaacat ccagagaagc acacaggttg     5520

gttttcttgg ctgccacgag cttgagcact cgagcggcaa aggcggactt gtggacgtta     5580

gctcgagctt cgtaggaggg cattttggtg gtgaagagga gactgaaata aatttagtct     5640

gcagaacttt ttatcggaac cttatctggg gcagtgaagt atatgttatg gtaatagtta     5700

cgagttagtt gaacttatag atagactgga ctatacggct atcggtccaa attagaaaga     5760

acgtcaatgg ctctctgggc gtcgcctttg ccgacaaaaa tgtgatcatg atgaaagcca     5820

gcaatgacgt tgcagctgat attgttgtcg gccaaccgcg ccgaaaacgc agctgtcaga     5880

cccacagcct ccaacgaaga atgtatcgtc aaagtgatcc aagcacactc atagttggag     5940

tcgtactcca aaggcggcaa tgacgagtca gacagatact cgtcgactca ggcgacgacg     6000

gaattcctgc agcccatctg cagaattcag gagagaccgg gttggcggcg tatttgtgtc     6060

ccaaaaaaca gccccaattg ccccggagaa gacggccagg ccgcctagat gacaaattca     6120

acaactcaca gctgactttc tgccattgcc actagggggg ggccttttta tatggccaag     6180

ccaagctctc cacgtcggtt gggctgcacc caacaataaa tgggtagggt tgcaccaaca     6240

aagggatggg atggggggta gaagatacga ggataacggg gctcaatggc acaaataaga     6300

acgaatactg ccattaagac tcgtgatcca gcgactgaca ccattgcatc atctaagggc     6360

ctcaaaacta cctcggaact gctgcgctga tctggacacc acagaggttc cgagcacttt     6420

aggttgcacc aaatgtccca ccaggtgcag gcagaaaacg ctggaacagc gtgtacagtt     6480

tgtcttaaca aaaagtgagg gcgctgaggt cgagcagggt ggtgtgactt gttatagcct     6540

ttagagctgc gaaagcgcgt atggatttgg ctcatcaggc cagattgagg gtctgtggac     6600

acatgtcatg ttagtgtact tcaatcgccc cctggatata gccccgacaa taggccgtgg     6660

cctcattttt ttgccttccg cacatttcca ttgctcggta cccacacctt gcttctcctg     6720

cacttgccaa ccttaatact ggtttacatt gaccaacatc ttacaagcgg ggggcttgtc     6780

tagggtatat ataaacagtg gctctcccaa tcggttgcca gtctcttttt tcctttcttt     6840

ccccacagat tcgaaatcta aactacacat cacacaatgc ctgttactga cgtccttaag     6900

cgaaagtccg gtgtcatcgt cggcgacgat gtccgagccg tgagtatcca cgacaagatc     6960

agtgtcgaga cgacgcgttt tgtgtaatga cacaatccga aagtcgctag caacacacac     7020

tctctacaca aactaaccca gctctccatg gctacaaccc gtctgtcggt tgagggcccc     7080

atctacgtgg gcttcgacgg cattcgtctt ctcaacctgc cgcatctcaa caagggctcg     7140

ggattccccc tcaacgagcg acgggaattc agactcagtg gtcttctgcc ctctgccgaa     7200

gccaccctgg aggaacaggt cgaccgagca taccaacaat tcaaaaagtg tggcactccc     7260

ttagccaaaa acgggttctg cacctcgctc aagttccaaa acgaggtgct ctactacgcc     7320

ctgctgctca agcacgttaa ggaggtcttc cccatcatct atacaccgac tcagggagaa     7380

gccattgaac agtactcgcg gctgttccgg cggcccgaag gctgcttcct cgacatcacc     7440

agtccctacg acgtggagga gcgtctggga gcgtttggag accatgacga cattgactac     7500

attgtcgtga ctgactccga gggtattctc ggaattggag accaaggagt gggcggtatt     7560

ggtatttcca tcgccaagct ggctctcatg actctatgtg ctggagtcaa cccctcacga     7620

gtcattcctg tggttctgga tacgggaacc aacaaccagg agctgctgca cgaccccctg     7680

tatctcggcc gacgaatgcc ccgagtgcga ggaaagcagt acgacgactt catcgacaac     7740

tttgtgcagt ctgcccgaag gctgtatccc aaggcggtga tccatttcga ggactttggg     7800

ctcgctaacg cacacaagat cctcgacaag tatcgaccgg agatcccctg cttcaacgac     7860

gacatccagg gcactggagc cgtcactttg gcctccatca cggccgctct caaggtgctg     7920

ggcaaaaata tcacagatac tcgaattctc gtgtacggag ctggttcggc cggcatgggt     7980

attgctgaac aggtctatga taacctggtt gcccagggtc tcgacgacaa gactgcgcga     8040

caaaacatct ttctcatgga ccgaccgggt ctactgacca ccgcacttac cgacgagcag     8100

atgagcgacg tgcagaagcc gtttgccaag gacaaggcca attacgaggg agtggacacc     8160

aagactctgg agcacgtggt tgctgccgtc aagccccata ttctcattgg atgttccact     8220

cagcccggcg cctttaacga gaaggtcgtc aaggagatgc tcaaacacac ccctcgaccc     8280

atcattctcc ctctttccaa ccccacacgt cttcatgagg ctgtccctgc agatctgtac     8340

aagtggaccg acggcaaggc tctggttgcc accggctcgc cctttgaccc agtcaacggc     8400

aaggagacgt ctgagaacaa taactgcttt gttttccccg gaatcgggct gggagccatt     8460

ctgtctcgat caaagctcat caccaacacc atgattgctg ctgccatcga gtgcctcgcc     8520

gaacaggccc ccattctcaa gaaccacgac gagggagtac ttcccgacgt agctctcatc     8580

cagatcattt cggcccgggt ggccactgcc gtggttcttc aggccaaggc tgagggccta     8640

gccactgtcg aggaagagct caagcccggc accaaggaac atgtgcagat tcccgacaac     8700

tttgacgagt gtctcgcctg ggtcgagact cagatgtggc ggcccgtcta ccggcctctc     8760

atccatgtgc gggattacga ctagagcgtg gaatagtgga atacagcttg ataacacagc     8820

gttgatcaga gtgagtccga gaagagtcca taggagtcca ttgaagagtc cattggagag     8880

tccattggag tccagtggcg gaccatagta gtcgacagta gttgatagta gtactggtag     8940

atgaatgtat tatcggcgcc agtgacagtt tagtttagta agcagc                    8986


<210>  11
<211>  31
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Primer ME-F

<400>  11
aaccatggta cggctacgaa ccatgcgacc c                                      31


<210>  12
<211>  34
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Primer ME-R

<400>  12
aagcggccgc ctagtcgtaa tcccgcacat ggat                                   34


<210>  13
<211>  8938
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Plasmid construct pME

<400>  13
gtacgggcgt cgttgcttgt gtgatttttg aggacccatc cctttggtat ataagtatac       60

tctggggtta aggttgcccg tgtagtctag gttatagttt tcatgtgaaa taccgagagc      120

cgagggagaa taaacggggg tatttggact tgtttttttc gcggaaaagc gtcgaatcaa      180

ccctgcgggc cttgcaccat gtccacgacg tgtttctcgc cccaattcgc cccttgcacg      240

tcaaaattag gcctccatct agacccctcc ataacatgtg actgtgggga aaagtataag      300

ggaaaccatg caaccataga cgacgtgaaa gacggggagg aaccaatgga ggccaaagaa      360

atggggtagc aacagtccag gagacagaca aggagacaag gagagggcgc ccgaaagatc      420

ggaaaaacaa acatgtccaa ttggggcagt gacggaaacg acacggacac ttcagtacaa      480

tggaccgacc atctccaagc cagggttatt ccggtatcac cttggccgta acctcccgct      540

ggtacctgat attgtacacg ttcacattca atatactttc agctacaata agagaggctg      600

tttgtcgggc atgtgtgtcc gtcgtatggg gtgatgtccg agggcgaaat tcgctacaag      660

cttaactctg gcgcttgtcc agtatgaata gacaagtcaa gaccagtggt gccatgattg      720

acagggaggt acaagacttc gatactcgag cattactcgg acttgtggcg attgaacaga      780

cgggcgatcg cttctccccc gtattgccgg cgcgccagct gcattaatga atcggccaac      840

gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc      900

tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt      960

tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg     1020

ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg     1080

agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat     1140

accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta     1200

ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct     1260

gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc     1320

ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa     1380

gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg     1440

taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag     1500

tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt     1560

gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta     1620

cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc     1680

agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca     1740

cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa     1800

cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat     1860

ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct     1920

taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt     1980

tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat     2040

ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta     2100

atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg     2160

gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt     2220

tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg     2280

cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg     2340

taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc     2400

ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa     2460

ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac     2520

cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt     2580

ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg     2640

gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa     2700

gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata     2760

aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgatgcg gtgtgaaata     2820

ccgcacagat gcgtaaggag aaaataccgc atcaggaaat tgtaagcgtt aatattttgt     2880

taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg     2940

gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt     3000

ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct     3060

atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt     3120

gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa     3180

agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc     3240

tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc     3300

tacagggcgc gtccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg     3360

ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg     3420

ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattgtaata     3480

cgactcacta tagggcgaat tgggcccgac gtcgcatgcg ctgatgacac tttggtctga     3540

aagagatgca ttttgaatcc caaacttgca gtgcccaagt gacatacatc tccgcgtttt     3600

ggaaaatgtt cagaaacagt tgattgtgtt ggaatgggga atggggaatg gaaaaatgac     3660

tcaagtatca attccaaaaa cttctctggc tggcagtacc tactgtccat actactgcat     3720

tttctccagt caggccactc tatactcgac gacacagtag taaaacccag ataatttcga     3780

cataaacaag aaaacagacc caataatatt tatatatagt cagccgtttg tccagttcag     3840

actgtaatag ccgaaaaaaa atccaaagtt tctattctag gaaaatatat tccaatattt     3900

ttaattctta atctcattta ttttattcta gcgaaataca tttcagctac ttgagacatg     3960

tgatacccac aaatcggatt cggactcggt tgttcagaag agcatatggc attcgtgctc     4020

gcttgttcac gtattcttcc tgttccatct cttggccgac aatcacacaa aaatggggtt     4080

ttttttttaa ttctaatgat tcattacagc aaaattgaga tatagcagac cacgtattcc     4140

ataatcacca aggaagttct tgggcgtctt aattaagtca tacacaagtc agctttcttc     4200

gagcctcata taagtataag tagttcaacg tattagcact gtacccagca tctccgtatc     4260

gagaaacaca acaacatgcc ccattggaca gatcatgcgg atacacaggt tgtgcagtat     4320

catacatact cgatcagaca ggtcgtctga ccatcataca agctgaacaa gcgctccata     4380

cttgcacgct ctctatatac acagttaaat tacatatcca tagtctaacc tctaacagtt     4440

aatcttctgg taagcctccc agccagcctt ctggtatcgc ttggcctcct caataggatc     4500

tcggttctgg ccgtacagac ctcggccgac aattatgata tccgttccgg tagacatgac     4560

atcctcaaca gttcggtact gctgtccgag agcgtctccc ttgtcgtcaa gacccacccc     4620

gggggtcaga ataagccagt cctcagagtc gcccttaggt cggttctggg caatgaagcc     4680

aaccacaaac tcggggtcgg atcgggcaag ctcaatggtc tgcttggagt actcgccagt     4740

ggccagagag cccttgcaag acagctcggc cagcatgagc agacctctgg ccagcttctc     4800

gttgggagag gggactagga actccttgta ctgggagttc tcgtagtcag agacgtcctc     4860

cttcttctgt tcagagacag tttcctcggc accagctcgc aggccagcaa tgattccggt     4920

tccgggtaca ccgtgggcgt tggtgatatc ggaccactcg gcgattcggt gacaccggta     4980

ctggtgcttg acagtgttgc caatatctgc gaactttctg tcctcgaaca ggaagaaacc     5040

gtgcttaaga gcaagttcct tgagggggag cacagtgccg gcgtaggtga agtcgtcaat     5100

gatgtcgata tgggttttga tcatgcacac ataaggtccg accttatcgg caagctcaat     5160

gagctccttg gtggtggtaa catccagaga agcacacagg ttggttttct tggctgccac     5220

gagcttgagc actcgagcgg caaaggcgga cttgtggacg ttagctcgag cttcgtagga     5280

gggcattttg gtggtgaaga ggagactgaa ataaatttag tctgcagaac tttttatcgg     5340

aaccttatct ggggcagtga agtatatgtt atggtaatag ttacgagtta gttgaactta     5400

tagatagact ggactatacg gctatcggtc caaattagaa agaacgtcaa tggctctctg     5460

ggcgtcgcct ttgccgacaa aaatgtgatc atgatgaaag ccagcaatga cgttgcagct     5520

gatattgttg tcggccaacc gcgccgaaaa cgcagctgtc agacccacag cctccaacga     5580

agaatgtatc gtcaaagtga tccaagcaca ctcatagttg gagtcgtact ccaaaggcgg     5640

caatgacgag tcagacagat actcgtcgac tcaggcgacg acggaattcc tgcagcccat     5700

ctgcagaatt caggagagac cgggttggcg gcgtatttgt gtcccaaaaa acagccccaa     5760

ttgccccgga gaagacggcc aggccgccta gatgacaaat tcaacaactc acagctgact     5820

ttctgccatt gccactaggg gggggccttt ttatatggcc aagccaagct ctccacgtcg     5880

gttgggctgc acccaacaat aaatgggtag ggttgcacca acaaagggat gggatggggg     5940

gtagaagata cgaggataac ggggctcaat ggcacaaata agaacgaata ctgccattaa     6000

gactcgtgat ccagcgactg acaccattgc atcatctaag ggcctcaaaa ctacctcgga     6060

actgctgcgc tgatctggac accacagagg ttccgagcac tttaggttgc accaaatgtc     6120

ccaccaggtg caggcagaaa acgctggaac agcgtgtaca gtttgtctta acaaaaagtg     6180

agggcgctga ggtcgagcag ggtggtgtga cttgttatag cctttagagc tgcgaaagcg     6240

cgtatggatt tggctcatca ggccagattg agggtctgtg gacacatgtc atgttagtgt     6300

acttcaatcg ccccctggat atagccccga caataggccg tggcctcatt tttttgcctt     6360

ccgcacattt ccattgctcg gtacccacac cttgcttctc ctgcacttgc caaccttaat     6420

actggtttac attgaccaac atcttacaag cggggggctt gtctagggta tatataaaca     6480

gtggctctcc caatcggttg ccagtctctt ttttcctttc tttccccaca gattcgaaat     6540

ctaaactaca catcacacaa tgcctgttac tgacgtcctt aagcgaaagt ccggtgtcat     6600

cgtcggcgac gatgtccgag ccgtgagtat ccacgacaag atcagtgtcg agacgacgcg     6660

ttttgtgtaa tgacacaatc cgaaagtcgc tagcaacaca cactctctac acaaactaac     6720

ccagctctcc atggtacgac tacgaaccat gcgacccaca cagaccagcg tcagggcggc     6780

gcttgggccc accgccgcgg cccgaaacat gtcctcctcc agcccctcca gcttcgaata     6840

ctcgtcctac gtcaagggca cgcgggaaat cggccaccga aaggcgccca caacccgtct     6900

gtcggttgag ggccccatct acgtgggctt cgacggcatt cgtcttctca acctgccgca     6960

tctcaacaag ggctcgggat tccccctcaa cgagcgacgg gaattcagac tcagtggtct     7020

tctgccctct gccgaagcca ccctggagga acaggtcgac cgagcatacc aacaattcaa     7080

aaagtgtggc actcccttag ccaaaaacgg gttctgcacc tcgctcaagt tccaaaacga     7140

ggtgctctac tacgccctgc tgctcaagca cgttaaggag gtcttcccca tcatctatac     7200

accgactcag ggagaagcca ttgaacagta ctcgcggctg ttccggcggc ccgaaggctg     7260

cttcctcgac atcaccagtc cctacgacgt ggaggagcgt ctgggagcgt ttggagacca     7320

tgacgacatt gactacattg tcgtgactga ctccgagggt attctcggaa ttggagacca     7380

aggagtgggc ggtattggta tttccatcgc caagctggct ctcatgactc tatgtgctgg     7440

agtcaacccc tcacgagtca ttcctgtggt tctggatacg ggaaccaaca accaggagct     7500

gctgcacgac cccctgtatc tcggccgacg aatgccccga gtgcgaggaa agcagtacga     7560

cgacttcatc gacaactttg tgcagtctgc ccgaaggctg tatcccaagg cggtgatcca     7620

tttcgaggac tttgggctcg ctaacgcaca caagatcctc gacaagtatc gaccggagat     7680

cccctgcttc aacgacgaca tccagggcac tggagccgtc actttggcct ccatcacggc     7740

cgctctcaag gtgctgggca aaaatatcac agatactcga attctcgtgt acggagctgg     7800

ttcggccggc atgggtattg ctgaacaggt ctatgataac ctggttgccc agggtctcga     7860

cgacaagact gcgcgacaaa acatctttct catggaccga ccgggtctac tgaccaccgc     7920

acttaccgac gagcagatga gcgacgtgca gaagccgttt gccaaggaca aggccaatta     7980

cgagggagtg gacaccaaga ctctggagca cgtggttgct gccgtcaagc cccatattct     8040

cattggatgt tccactcagc ccggcgcctt taacgagaag gtcgtcaagg agatgctcaa     8100

acacacccct cgacccatca ttctccctct ttccaacccc acacgtcttc atgaggctgt     8160

ccctgcagat ctgtacaagt ggaccgacgg caaggctctg gttgccaccg gctcgccctt     8220

tgacccagtc aacggcaagg agacgtctga gaacaataac tgctttgttt tccccggaat     8280

cgggctggga gccattctgt ctcgatcaaa gctcatcacc aacaccatga ttgctgctgc     8340

catcgagtgc ctcgccgaac aggcccccat tctcaagaac cacgacgagg gagtacttcc     8400

cgacgtagct ctcatccaga tcatttcggc ccgggtggcc actgccgtgg ttcttcaggc     8460

caaggctgag ggcctagcca ctgtcgagga agagctcaag cccggcacca aggaacatgt     8520

gcagattccc gacaactttg acgagtgtct cgcctgggtc gagactcaga tgtggcggcc     8580

cgtctaccgg cctctcatcc atgtgcggga ttacgactag cggccgcaag tgtggatggg     8640

gaagtgagtg cccggttctg tgtgcacaat tggcaatcca agatggatgg attcaacaca     8700

gggatatagc gagctacgtg gtggtgcgag gatatagcaa cggatattta tgtttgacac     8760

ttgagaatgt acgatacaag cactgtccaa gtacaatact aaacatactg tacatactca     8820

tactcgtacc cgggcaacgg tttcacttga gtgcagtggc tagtgctctt actcgtacag     8880

tgtgcaatac tgcgtatcat agtctttgat gtatatcgta ttcattcatg ttagttgc       8938


<210>  14
<211>  4604
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Plasmid construct pBlue-YURA3

<400>  14
ggccgctcta gaactagtgg atcccccggg ctgcaggaat tcctttacct gcaggataac       60

ttcgtataat gtatgctata cgaagttatg atctctctct tgagcttttc cataacaagt      120

tcttctgcct ccaggaagtc catgggtggt ttgatcatgg ttttggtgta gtggtagtgc      180

agtggtggta ttgtgactgg ggatgtagtt gagaataagt catacacaag tcagctttct      240

tcgagcctca tataagtata agtagttcaa cgtattagca ctgtacccag catctccgta      300

tcgagaaaca caacaacatg ccccattgga cagatcatgc ggatacacag gttgtgcagt      360

atcatacata ctcgatcaga caggtcgtct gaccatcata caagctgaac aagcgctcca      420

tacttgcacg ctctctatat acacagttaa attacatatc catagtctaa cctctaacag      480

ttaatcttct ggtaagcctc ccagccagcc ttctggtatc gcttggcctc ctcaatagga      540

tctcggttct ggccgtacag acctcggccg acaattatga tatccgttcc ggtagacatg      600

acatcctcaa cagttcggta ctgctgtccg agagcgtctc ccttgtcgtc aagacccacc      660

ccgggggtca gaataagcca gtcctcagag tcgcccttag gtcggttctg ggcaatgaag      720

ccaaccacaa actcggggtc ggatcgggca agctcaatgg tctgcttgga gtactcgcca      780

gtggccagag agcccttgca agacagctcg gccagcatga gcagacctct ggccagcttc      840

tcgttgggag aggggactag gaactccttg tactgggagt tctcgtagtc agagacgtcc      900

tccttcttct gttcagagac agtttcctcg gcaccagctc gcaggccagc aatgattccg      960

gttccgggta caccgtgggc gttggtgata tcggaccact cggcgattcg gtgacaccgg     1020

tactggtgct tgacagtgtt gccaatatct gcgaactttc tgtcctcgaa caggaagaaa     1080

ccgtgcttaa gagcaagttc cttgaggggg agcacagtgc cggcgtaggt gaagtcgtca     1140

atgatgtcga tatgggtttt gatcatgcac acataaggtc cgaccttatc ggcaagctca     1200

atgagctcct tggtggtggt aacatccaga gaagcacaca ggttggtttt cttggctgcc     1260

acgagcttga gcactcgagc ggcaaaggcg gacttgtgga cgttagctcg agcttcgtag     1320

gagggcattt tggtggtgaa gaggagactg aaataaattt agtctgcaga actttttatc     1380

ggaaccttat ctggggcagt gaagtatatg ttatggtaat agttacgagt tagttgaact     1440

tatagataga ctggactata cggctatcgg tccaaattag aaagaacgtc aatggctctc     1500

tgggcgtcgc ctttgccgac aaaaatgtga tcatgatgaa agccagcaat gacgttgcag     1560

ctgatattgt tgtcggccaa ccgcgccgaa aacgcagctg tcagacccac agcctccaac     1620

gaagaatgta tcgtcaaagt gatccaagca cactcatagt tggagtcgta ctccaaaggc     1680

ggcaatgacg agtcagacag atactcgtcg acctcgaggg ggggcccggt acccaattcg     1740

ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg     1800

gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg     1860

cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc     1920

gaatggaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca     1980

gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga     2040

ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg     2100

actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat     2160

caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag     2220

ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga     2280

agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa     2340

ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg     2400

gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc     2460

tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta     2520

ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg     2580

ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg     2640

gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac     2700

gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg     2760

acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt     2820

actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg     2880

ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac     2940

cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt     3000

gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag     3060

caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc     3120

aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc     3180

ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta     3240

tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg     3300

ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga     3360

ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac     3420

ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa     3480

tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat     3540

cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc     3600

taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg     3660

gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc     3720

acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg     3780

ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg     3840

ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa     3900

cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg     3960

aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga     4020

gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct     4080

gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca     4140

gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc     4200

ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg     4260

ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc     4320

caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca     4380

ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc     4440

attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga     4500

gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta     4560

accctcacta aagggaacaa aagctggagc tccaccgcgg tggc                      4604


<210>  15
<211>  6728
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Plasmid construct pME-KO

<400>  15
aattccttta cctgcaggat aacttcgtat aatgtatgct atacgaagtt atgatctctc       60

tcttgagctt ttccataaca agttcttctg cctccaggaa gtccatgggt ggtttgatca      120

tggttttggt gtagtggtag tgcagtggtg gtattgtgac tggggatgta gttgagaata      180

agtcatacac aagtcagctt tcttcgagcc tcatataagt ataagtagtt caacgtatta      240

gcactgtacc cagcatctcc gtatcgagaa acacaacaac atgccccatt ggacagatca      300

tgcggataca caggttgtgc agtatcatac atactcgatc agacaggtcg tctgaccatc      360

atacaagctg aacaagcgct ccatacttgc acgctctcta tatacacagt taaattacat      420

atccatagtc taacctctaa cagttaatct tctggtaagc ctcccagcca gccttctggt      480

atcgcttggc ctcctcaata ggatctcggt tctggccgta cagacctcgg ccgacaatta      540

tgatatccgt tccggtagac atgacatcct caacagttcg gtactgctgt ccgagagcgt      600

ctcccttgtc gtcaagaccc accccggggg tcagaataag ccagtcctca gagtcgccct      660

taggtcggtt ctgggcaatg aagccaacca caaactcggg gtcggatcgg gcaagctcaa      720

tggtctgctt ggagtactcg ccagtggcca gagagccctt gcaagacagc tcggccagca      780

tgagcagacc tctggccagc ttctcgttgg gagaggggac taggaactcc ttgtactggg      840

agttctcgta gtcagagacg tcctccttct tctgttcaga gacagtttcc tcggcaccag      900

ctcgcaggcc agcaatgatt ccggttccgg gtacaccgtg ggcgttggtg atatcggacc      960

actcggcgat tcggtgacac cggtactggt gcttgacagt gttgccaata tctgcgaact     1020

ttctgtcctc gaacaggaag aaaccgtgct taagagcaag ttccttgagg gggagcacag     1080

tgccggcgta ggtgaagtcg tcaatgatgt cgatatgggt tttgatcatg cacacataag     1140

gtccgacctt atcggcaagc tcaatgagct ccttggtggt ggtaacatcc agagaagcac     1200

acaggttggt tttcttggct gccacgagct tgagcactcg agcggcaaag gcggacttgt     1260

ggacgttagc tcgagcttcg taggagggca ttttggtggt gaagaggaga ctgaaataaa     1320

tttagtctgc agaacttttt atcggaacct tatctggggc agtgaagtat atgttatggt     1380

aatagttacg agttagttga acttatagat agactggact atacggctat cggtccaaat     1440

tagaaagaac gtcaatggct ctctgggcgt cgcctttgcc gacaaaaatg tgatcatgat     1500

gaaagccagc aatgacgttg cagctgatat tgttgtcggc caaccgcgcc gaaaacgcag     1560

ctgtcagacc cacagcctcc aacgaagaat gtatcgtcaa agtgatccaa gcacactcat     1620

agttggagtc gtactccaaa ggcggcaatg acgagtcaga cagatactcg tcgacctcga     1680

gggggggccc ggtacccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg     1740

tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag     1800

cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc     1860

aacagttgcg cagcctgaat ggcgaatgga aattgtaagc gttaatattt tgttaaaatt     1920

cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat     1980

cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa     2040

gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg     2100

cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa     2160

agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc     2220

gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag     2280

tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg     2340

cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta     2400

aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata     2460

ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc     2520

ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga     2580

agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct     2640

tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg     2700

tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta     2760

ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat     2820

gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt     2880

acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga     2940

tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga     3000

gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga     3060

actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc     3120

aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc     3180

cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg     3240

tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat     3300

cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata     3360

tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct     3420

ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga     3480

ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg     3540

cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc     3600

aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct     3660

agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc     3720

tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt     3780

ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg     3840

cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct     3900

atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag     3960

ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag     4020

tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg     4080

gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg     4140

gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac     4200

cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt     4260

gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat     4320

tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc     4380

aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc     4440

tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca     4500

tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg agctccaccg     4560

cggtggcggc cgctctagaa ctagtggatc cgtgcccacg tttcgtttct tcaccggcgg     4620

cacatccacg cggtcctccg gcccgccgcg ggcctcccct agcttctgtg cttcctcgtc     4680

gcgcttcatc tggtgaaaac ggggtttttt gacactcgat ttttcgacac tcgatttttc     4740

gacacttgat ttttcagcga acccagatta tgcgctacac cagcgcctgc atatgcggcg     4800

ttagtgggga agtggagtag ttggagccgt gtatgatact agagtggcgc tgagggcgcg     4860

tttggtggtc gttccggttg ccctatgaga ctgaacctgg tttcgacagg tgttttgcac     4920

tcattcgcct acctctcgct tatcaccgat ttatgtttca tttcattgga ctattgttag     4980

tttttccgcc aaaactcaaa atgttttctg acatctgact agaacacggc tcctgtcaat     5040

gagcaaggat tgagtgtcta caagtgcaag gagtggtagt gttcaatggc ttttggtcta     5100

gaaccagatg agcatgaaaa aaggaacgtg agtttcctga tattcccgcc gctaatcatt     5160

ctgtctctct tacccaccca aagcgtccct ccgatgacat ttccatacga gtgcttacta     5220

aactaaactg tcactggcgc cgataataca ttcatctacc agtactacta tcaactactg     5280

tcgactacta tggtccgcca ctggactcca atggactctc caatggactc ttcaatggac     5340

tcctatggac tcttctcgga ctcactctga tcaacgctgt gttatcaagc tgtattccac     5400

tattccacgc tctagtcgta atcccgcaca tggatgagag gccggtagac gggccgccac     5460

atctgagtct cgacccaggc gagacactcg tcaaagttgt cgggaatctg cacatgttcc     5520

ttggtgccgg gcttgagctc ttcctcgaca gtggctaggc cctcagcctt ggcctgaaga     5580

accacggcag tggccacccg ggccgaaatg atctggatga gagctacgtc gggaagtact     5640

ccctcgtcgt ggttcttgag aatgggggcc tgttcggcga ggcactcgat ggcagcagca     5700

atcatggtgt tggtgatgag ctttgatcga gacagaatgc tcgaggacgt aggacgagta     5760

ttcgaagctg gaggggctgg aggaggacat gtttcgggcc gcggcggtgg gcccaagcgc     5820

cgccctgacg ctggtctgtg tgggtcgcat ggttcgtagt cgtaacatgg cgaatatcag     5880

cgtgtacagg agtacttgta tactggaagc agtatcttgg caggaattta aatcgccccc     5940

ttaagagttc gacaacactc acaacctctc aacctagatt ggctatcaac cttgcgggat     6000

tatgcagcgc gacagggagc ataaacgggg tgttatgggg gctgaaaacg gctggaaaag     6060

acataaatcg gacaattggc ggccagaaaa tgttgcgggg aaaaaaccgc cccgaaaacc     6120

gccctgctcg tccaattgac cgcatcttct gtccccactg acgccttcca tcctgcaggc     6180

caagacaacg acatgcacgc acgtggaaca ggcagaccta aaatgagggt tgtaggatat     6240

taaagctagt gatttacgac caagaaacat gtagagtacg agtatagtac gtagaacaat     6300

gatgctccaa atagagagaa gatggcatgt cacttggtac ggccatggaa tgcacgcatt     6360

tcctcaatat ggttagtttt taacattgac agcactagtc atgcaccata tcatatcagc     6420

ccacatggtg agacacacat cgcatgcaca ggtcaggttc tgggaggtga tgaataaata     6480

ataatacaat aaataagaaa aatgcaacta aatgaatctg cgctggattt cctccaaatc     6540

cccctgtagc tccttatgtc gtttgatgtt gcgagaaaca atgttcttga tgctcaaggt     6600

cagctcgtgc tcatggacct ggtagaagcc gccaagctcg gaggccaagg tgatctgggc     6660

gttcttggtc tccatcgaca gtcgtctgat ttcgttctcc gtgatgtgga tacggttttt     6720

gatcagcg                                                              6728


<210>  16
<211>  34
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Primer YME-5-1

<400>  16
gatcaagaat tcgctgatca aaaaccgtat ccac                                   34


<210>  17
<211>  34
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Primer YME-5-2

<400>  17
gatcaactcg aggacgtagg acgagtattc gaag                                   34


<210>  18
<211>  34
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Primer YME-3-1

<400>  18
gatcaactcg agcattctgt ctcgatcaaa gctc                                   34


<210>  19
<211>  35
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Primer YME-3-2

<400>  19
gatcaaggat ccgtgcccac gtttcgtttc ttcac                                  35


<210>  20
<211>  23
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Primer YME-5-confirm-1

<400>  20
ctcaaatctc gcatttgaga ctc                                               23


<210>  21
<211>  21
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Primer YME-5-confirm-2

<400>  21
gcagcagcaa tcatggtgtt g                                                 21


<210>  22
<211>  22
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Primer YME-3-confirm-1

<400>  22
gagtgttgtc gaactcttaa gg                                                22


<210>  23
<211>  22
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Primer YME-3-confirm-2

<400>  23
gttgtgttga gcacgtggca tc                                                22


<210>  24
<211>  1017
<212>  DNA
<213>  Yarrowia lipolytica

<400>  24
atgttccgaa cccgagttac cggctccacc ctgcgatcct tctccacctc cgctgcccga       60

cagcacaagg ttgtcgtcct tggcgccaac ggaggcattg gccagcccct gtctctgctg      120

ctcaagctca acaagaacgt gaccgacctc ggtctgtacg atctgcgagg cgcccccggc      180

gttgctgccg atgtctccca catccccacc aactccaccg tggccggcta ctctcccgac      240

aacaacggca ttgccgaggc cctcaagggc gccaagctgg tgctgatccc cgccggtgtc      300

ccccgaaagc ccggcatgac ccgagacgat ctgttcaaca ccaacgcctc cattgtgcga      360

gacctggcca aggccgtcgg tgagcacgcc cccgacgcct ttgtcggagt cattgctaac      420

cccgtcaact ccaccgtccc cattgtcgcc gaggtgctca agtccaaggg caagtacgac      480

cccaagaagc tcttcggtgt caccaccctc gacgtcatcc gagccgagcg attcgtctcc      540

cagctcgagc acaccaaccc caccaaggag tacttccccg ttgttggcgg ccactccggt      600

gtcaccattg tccccctcgt gtcccagtcc gaccaccccg acattgccgg tgaggctcga      660

gacaagcttg tccaccgaat ccagtttggc ggtgacgagg ttgtcaaggc caaggacggt      720

gccggatccg ccaccctttc catggcccag gctgccgccc gattcgccga ctctctcctc      780

cgaggtgtca acggcgagaa ggacgttgtt gagcccactt tcgtcgactc tcctctgttc      840

aagggtgagg gcatcgactt cttctccacc aaggtcactc ttggccctaa cggtgttgag      900

gagatccacc ccatcggaaa ggtcaacgag tacgaggaga agctcatcga ggctgccaag      960

gccgatctca agaagaacat tgagaagggt gtcaactttg tcaagcagaa cccttaa        1017


<210>  25
<211>  338
<212>  PRT
<213>  Yarrowia lipolytica

<400>  25

Met Phe Arg Thr Arg Val Thr Gly Ser Thr Leu Arg Ser Phe Ser Thr 
1               5                   10                  15      


Ser Ala Ala Arg Gln His Lys Val Val Val Leu Gly Ala Asn Gly Gly 
            20                  25                  30          


Ile Gly Gln Pro Leu Ser Leu Leu Leu Lys Leu Asn Lys Asn Val Thr 
        35                  40                  45              


Asp Leu Gly Leu Tyr Asp Leu Arg Gly Ala Pro Gly Val Ala Ala Asp 
    50                  55                  60                  


Val Ser His Ile Pro Thr Asn Ser Thr Val Ala Gly Tyr Ser Pro Asp 
65                  70                  75                  80  


Asn Asn Gly Ile Ala Glu Ala Leu Lys Gly Ala Lys Leu Val Leu Ile 
                85                  90                  95      


Pro Ala Gly Val Pro Arg Lys Pro Gly Met Thr Arg Asp Asp Leu Phe 
            100                 105                 110         


Asn Thr Asn Ala Ser Ile Val Arg Asp Leu Ala Lys Ala Val Gly Glu 
        115                 120                 125             


His Ala Pro Asp Ala Phe Val Gly Val Ile Ala Asn Pro Val Asn Ser 
    130                 135                 140                 


Thr Val Pro Ile Val Ala Glu Val Leu Lys Ser Lys Gly Lys Tyr Asp 
145                 150                 155                 160 


Pro Lys Lys Leu Phe Gly Val Thr Thr Leu Asp Val Ile Arg Ala Glu 
                165                 170                 175     


Arg Phe Val Ser Gln Leu Glu His Thr Asn Pro Thr Lys Glu Tyr Phe 
            180                 185                 190         


Pro Val Val Gly Gly His Ser Gly Val Thr Ile Val Pro Leu Val Ser 
        195                 200                 205             


Gln Ser Asp His Pro Asp Ile Ala Gly Glu Ala Arg Asp Lys Leu Val 
    210                 215                 220                 


His Arg Ile Gln Phe Gly Gly Asp Glu Val Val Lys Ala Lys Asp Gly 
225                 230                 235                 240 


Ala Gly Ser Ala Thr Leu Ser Met Ala Gln Ala Ala Ala Arg Phe Ala 
                245                 250                 255     


Asp Ser Leu Leu Arg Gly Val Asn Gly Glu Lys Asp Val Val Glu Pro 
            260                 265                 270         


Thr Phe Val Asp Ser Pro Leu Phe Lys Gly Glu Gly Ile Asp Phe Phe 
        275                 280                 285             


Ser Thr Lys Val Thr Leu Gly Pro Asn Gly Val Glu Glu Ile His Pro 
    290                 295                 300                 


Ile Gly Lys Val Asn Glu Tyr Glu Glu Lys Leu Ile Glu Ala Ala Lys 
305                 310                 315                 320 


Ala Asp Leu Lys Lys Asn Ile Glu Lys Gly Val Asn Phe Val Lys Gln 
                325                 330                 335     


Asn Pro 
        


<210>  26
<211>  996
<212>  DNA
<213>  Yarrowia lipolytica

<400>  26
atggttaaag ctgtcgttgc cggagccgct ggtggtattg gccagcccct ttctcttctc       60

ctcaaactct ctccttacgt gaccgagctt gctctctacg atgtcgtcaa ctcccccggt      120

gttgccgctg acctctccca catctccacc aaggctaagg tcactggcta cctccccaag      180

gatgacggtc tcaagaacgc tctgaccggc gccaacattg tcgttatccc cgccggtatc      240

ccccgaaagc ccggtatgac ccgagacgat ctgttcaaga tcaacgctgg tatcgtccga      300

gatctcgtca ccggtgtcgc ccagtacgcc cctgacgcct ttgtgctcat catctccaac      360

cccgtcaact ctaccgtccc tattgctgcc gaggtcctca agaagcacaa cgtcttcaac      420

cctaagaagc tcttcggtgt caccaccctt gacgttgtcc gagcccagac cttcaccgcc      480

gctgttgttg gcgagtctga ccccaccaag ctcaacatcc ccgtcgttgg tggccactcc      540

ggagacacca ttgtccctct cctgtctctg accaagccta aggtcgagat ccccgccgac      600

aagctcgacg acctcgtcaa gcgaatccag tttggtggtg acgaggttgt ccaggctaag      660

gacggtcttg gatccgctac cctctccatg gcccaggctg gtttccgatt tgccgaggct      720

gtcctcaagg gtgccgctgg tgagaagggc atcatcgagc ccgcctacat ctaccttgac      780

ggtattgatg gcacctccga catcaagcga gaggtcggtg tcgccttctt ctctgtccct      840

gtcgagttcg gccctgaggg tgccgctaag gcttacaaca tccttcccga ggccaacgac      900

tacgagaaga agcttctcaa ggtctccatc gacggtcttt acggcaacat tgccaagggc      960

gaggagttca ttgttaaccc tcctcctgcc aactag                                996


<210>  27
<211>  331
<212>  PRT
<213>  Yarrowia lipolytica

<400>  27

Met Val Lys Ala Val Val Ala Gly Ala Ala Gly Gly Ile Gly Gln Pro 
1               5                   10                  15      


Leu Ser Leu Leu Leu Lys Leu Ser Pro Tyr Val Thr Glu Leu Ala Leu 
            20                  25                  30          


Tyr Asp Val Val Asn Ser Pro Gly Val Ala Ala Asp Leu Ser His Ile 
        35                  40                  45              


Ser Thr Lys Ala Lys Val Thr Gly Tyr Leu Pro Lys Asp Asp Gly Leu 
    50                  55                  60                  


Lys Asn Ala Leu Thr Gly Ala Asn Ile Val Val Ile Pro Ala Gly Ile 
65                  70                  75                  80  


Pro Arg Lys Pro Gly Met Thr Arg Asp Asp Leu Phe Lys Ile Asn Ala 
                85                  90                  95      


Gly Ile Val Arg Asp Leu Val Thr Gly Val Ala Gln Tyr Ala Pro Asp 
            100                 105                 110         


Ala Phe Val Leu Ile Ile Ser Asn Pro Val Asn Ser Thr Val Pro Ile 
        115                 120                 125             


Ala Ala Glu Val Leu Lys Lys His Asn Val Phe Asn Pro Lys Lys Leu 
    130                 135                 140                 


Phe Gly Val Thr Thr Leu Asp Val Val Arg Ala Gln Thr Phe Thr Ala 
145                 150                 155                 160 


Ala Val Val Gly Glu Ser Asp Pro Thr Lys Leu Asn Ile Pro Val Val 
                165                 170                 175     


Gly Gly His Ser Gly Asp Thr Ile Val Pro Leu Leu Ser Leu Thr Lys 
            180                 185                 190         


Pro Lys Val Glu Ile Pro Ala Asp Lys Leu Asp Asp Leu Val Lys Arg 
        195                 200                 205             


Ile Gln Phe Gly Gly Asp Glu Val Val Gln Ala Lys Asp Gly Leu Gly 
    210                 215                 220                 


Ser Ala Thr Leu Ser Met Ala Gln Ala Gly Phe Arg Phe Ala Glu Ala 
225                 230                 235                 240 


Val Leu Lys Gly Ala Ala Gly Glu Lys Gly Ile Ile Glu Pro Ala Tyr 
                245                 250                 255     


Ile Tyr Leu Asp Gly Ile Asp Gly Thr Ser Asp Ile Lys Arg Glu Val 
            260                 265                 270         


Gly Val Ala Phe Phe Ser Val Pro Val Glu Phe Gly Pro Glu Gly Ala 
        275                 280                 285             


Ala Lys Ala Tyr Asn Ile Leu Pro Glu Ala Asn Asp Tyr Glu Lys Lys 
    290                 295                 300                 


Leu Leu Lys Val Ser Ile Asp Gly Leu Tyr Gly Asn Ile Ala Lys Gly 
305                 310                 315                 320 


Glu Glu Phe Ile Val Asn Pro Pro Pro Ala Asn 
                325                 330     


<210>  28
<211>  999
<212>  DNA
<213>  Yarrowia lipolytica

<400>  28
atggttaaag ctgtcgttgc cggagccgct ggtggtattg gccagcccct ttctcttctc       60

ctcaaactct ctccttacgt gaccgagctt gctctctacg atgtcgtcaa ctcccccggt      120

gttgccgctg acctctccca catctccacc aaggctaagg tcactggcta cctccccaag      180

gatgacggtc tcaagaacgc tctgaccggc gccaacattg tcgttatccc cgccggtatc      240

ccccgaaagc ccggtatgac ccgagacgat ctgttcaaga tcaacgctgg tatcgtccga      300

gatctcgtca ccggtgtcgc ccagtacgcc cctgacgcct ttgtgctcat catctccaac      360

cccgtcaact ctaccgtccc tattgctgcc gaggtcctca agaagcacaa cgtcttcaac      420

cctaagaagc tcttcggtgt caccaccctt gacgttgtcc gagcccagac cttcaccgcc      480

gctgttgttg gcgagtctga ccccaccaag ctcaacatcc ccgtcgttgg tggccactcc      540

ggagacacca ttgtccctct cctgtctctg accaagccta aggtcgagat ccccgccgac      600

aagctcgacg acctcgtcaa gcgaatccag tttggtggtg acgaggttgt ccaggctaag      660

gacggtcttg gatccgctac cctctccatg gcccaggctg gtttccgatt tgccgaggct      720

gtcctcaagg gtgccgctgg tgagaagggc atcatcgagc ccgcctacat ctaccttgac      780

ggtattgatg gcacctccga catcaagcga gaggtcggtg tcgccttctt ctctgtccct      840

gtcgagttcg gccctgaggg tgccgctaag gcttacaaca tccttcccga ggccaacgac      900

tacgagaaga agcttctcaa ggtctccatc gacggtcttt acggcaacat tgccaagggc      960

gaggagttca ttgttaaccc tcctcctgcc aagatctaa                             999


<210>  29
<211>  332
<212>  PRT
<213>  Yarrowia lipolytica

<400>  29

Met Val Lys Ala Val Val Ala Gly Ala Ala Gly Gly Ile Gly Gln Pro 
1               5                   10                  15      


Leu Ser Leu Leu Leu Lys Leu Ser Pro Tyr Val Thr Glu Leu Ala Leu 
            20                  25                  30          


Tyr Asp Val Val Asn Ser Pro Gly Val Ala Ala Asp Leu Ser His Ile 
        35                  40                  45              


Ser Thr Lys Ala Lys Val Thr Gly Tyr Leu Pro Lys Asp Asp Gly Leu 
    50                  55                  60                  


Lys Asn Ala Leu Thr Gly Ala Asn Ile Val Val Ile Pro Ala Gly Ile 
65                  70                  75                  80  


Pro Arg Lys Pro Gly Met Thr Arg Asp Asp Leu Phe Lys Ile Asn Ala 
                85                  90                  95      


Gly Ile Val Arg Asp Leu Val Thr Gly Val Ala Gln Tyr Ala Pro Asp 
            100                 105                 110         


Ala Phe Val Leu Ile Ile Ser Asn Pro Val Asn Ser Thr Val Pro Ile 
        115                 120                 125             


Ala Ala Glu Val Leu Lys Lys His Asn Val Phe Asn Pro Lys Lys Leu 
    130                 135                 140                 


Phe Gly Val Thr Thr Leu Asp Val Val Arg Ala Gln Thr Phe Thr Ala 
145                 150                 155                 160 


Ala Val Val Gly Glu Ser Asp Pro Thr Lys Leu Asn Ile Pro Val Val 
                165                 170                 175     


Gly Gly His Ser Gly Asp Thr Ile Val Pro Leu Leu Ser Leu Thr Lys 
            180                 185                 190         


Pro Lys Val Glu Ile Pro Ala Asp Lys Leu Asp Asp Leu Val Lys Arg 
        195                 200                 205             


Ile Gln Phe Gly Gly Asp Glu Val Val Gln Ala Lys Asp Gly Leu Gly 
    210                 215                 220                 


Ser Ala Thr Leu Ser Met Ala Gln Ala Gly Phe Arg Phe Ala Glu Ala 
225                 230                 235                 240 


Val Leu Lys Gly Ala Ala Gly Glu Lys Gly Ile Ile Glu Pro Ala Tyr 
                245                 250                 255     


Ile Tyr Leu Asp Gly Ile Asp Gly Thr Ser Asp Ile Lys Arg Glu Val 
            260                 265                 270         


Gly Val Ala Phe Phe Ser Val Pro Val Glu Phe Gly Pro Glu Gly Ala 
        275                 280                 285             


Ala Lys Ala Tyr Asn Ile Leu Pro Glu Ala Asn Asp Tyr Glu Lys Lys 
    290                 295                 300                 


Leu Leu Lys Val Ser Ile Asp Gly Leu Tyr Gly Asn Ile Ala Lys Gly 
305                 310                 315                 320 


Glu Glu Phe Ile Val Asn Pro Pro Pro Ala Lys Ile 
                325                 330         


<210>  30
<211>  338
<212>  PRT
<213>  Yarrowia lipolytica

<400>  30

Met Val Arg Thr Arg Val Thr Gly Ser Thr Leu Arg Ser Phe Ser Thr 
1               5                   10                  15      


Ser Ala Ala Arg Gln His Lys Val Val Val Leu Gly Ala Asn Gly Gly 
            20                  25                  30          


Ile Gly Gln Pro Leu Ser Leu Leu Leu Lys Leu Asn Lys Asn Val Thr 
        35                  40                  45              


Asp Leu Gly Leu Tyr Asp Leu Arg Gly Ala Pro Gly Val Ala Ala Asp 
    50                  55                  60                  


Val Ser His Ile Pro Thr Asn Ser Thr Val Ala Gly Tyr Ser Pro Asp 
65                  70                  75                  80  


Asn Asn Gly Ile Ala Glu Ala Leu Lys Gly Ala Lys Leu Val Leu Ile 
                85                  90                  95      


Pro Ala Gly Val Pro Arg Lys Pro Gly Met Thr Arg Asp Asp Leu Phe 
            100                 105                 110         


Asn Thr Asn Ala Ser Ile Val Arg Asp Leu Ala Lys Ala Val Gly Glu 
        115                 120                 125             


His Ala Pro Asp Ala Phe Val Gly Val Ile Ala Asn Pro Val Asn Ser 
    130                 135                 140                 


Thr Val Pro Ile Val Ala Glu Val Leu Lys Ser Lys Gly Lys Tyr Asp 
145                 150                 155                 160 


Pro Lys Lys Leu Phe Gly Val Thr Thr Leu Asp Val Ile Arg Ala Glu 
                165                 170                 175     


Arg Phe Val Ser Gln Leu Glu His Thr Asn Pro Thr Lys Glu Tyr Phe 
            180                 185                 190         


Pro Val Val Gly Gly His Ser Gly Val Thr Ile Val Pro Leu Val Ser 
        195                 200                 205             


Gln Ser Asp His Pro Asp Ile Ala Gly Glu Ala Arg Asp Lys Leu Val 
    210                 215                 220                 


His Arg Ile Gln Phe Gly Gly Asp Glu Val Val Lys Ala Lys Asp Gly 
225                 230                 235                 240 


Ala Gly Ser Ala Thr Leu Ser Met Ala Gln Ala Ala Ala Arg Phe Ala 
                245                 250                 255     


Asp Ser Leu Leu Arg Gly Val Asn Gly Glu Lys Asp Val Val Glu Pro 
            260                 265                 270         


Thr Phe Val Asp Ser Pro Leu Phe Lys Gly Glu Gly Ile Asp Phe Phe 
        275                 280                 285             


Ser Thr Lys Val Thr Leu Gly Pro Asn Gly Val Glu Glu Ile His Pro 
    290                 295                 300                 


Ile Gly Lys Val Asn Glu Tyr Glu Glu Lys Leu Ile Glu Ala Ala Lys 
305                 310                 315                 320 


Ala Asp Leu Lys Lys Asn Ile Glu Lys Gly Val Asn Phe Val Lys Gln 
                325                 330                 335     


Asn Pro 
        


<210>  31
<211>  32
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Primer YMDH1-F

<400>  31
gatcaaccat ggtccgaacc cgagttaccg gc                                     32


<210>  32
<211>  36
<212>  DNA
<213>  Artificial sequence

<220>
<223>  Primer YMDH1-R

<400>  32
gatcaagcgg ccgcttaagg gttctgcttg acaaag                                 36


