
                         SEQUENCE LISTING

<110>  Evonik Degussa GmbH
 
<120>  Method for producing L-amino acids

<130>  201600130

<160>  14    

<170>  PatentIn version 3.5

<210>  1
<211>  1719
<212>  DNA
<213>  Saccharomyces cerevisiae


<220>
<221>  misc_feature
<222>  (1)..(6)
<223>  NdeI restriction site

<220>
<221>  CDS
<222>  (4)..(1719)
<223>  sequence of PDC1 of Sacharomyces cerevisiae optimized for the 
       codon usage of Escherichia coli with a carboxy-terminal His6-tag

<220>
<221>  misc_feature
<222>  (1693)..(1698)
<223>  XhoI restriction site

<400>  1
cat atg agc gaa att acc ctg ggc aaa tac ctg ttt gaa cgc ctg aaa         48
    Met Ser Glu Ile Thr Leu Gly Lys Tyr Leu Phe Glu Arg Leu Lys           
    1               5                   10                  15            

cag gtt aat gtg aat acc gtt ttt ggt ctg cct ggc gat ttt aat ctg         96
Gln Val Asn Val Asn Thr Val Phe Gly Leu Pro Gly Asp Phe Asn Leu           
                20                  25                  30                

agc ctg ctg gat aaa atc tat gaa gtt gaa ggt atg cgt tgg gca ggt        144
Ser Leu Leu Asp Lys Ile Tyr Glu Val Glu Gly Met Arg Trp Ala Gly           
            35                  40                  45                    

aat gca aat gaa ctg aat gca gcc tat gca gca gat ggt tat gca cgt        192
Asn Ala Asn Glu Leu Asn Ala Ala Tyr Ala Ala Asp Gly Tyr Ala Arg           
        50                  55                  60                        

att aaa ggt atg agc tgc att att acc acc ttt ggt gtt ggt gaa ctg        240
Ile Lys Gly Met Ser Cys Ile Ile Thr Thr Phe Gly Val Gly Glu Leu           
    65                  70                  75                            

agc gca ctg aat ggt att gca ggt agc tat gca gaa cat gtt ggt gtg        288
Ser Ala Leu Asn Gly Ile Ala Gly Ser Tyr Ala Glu His Val Gly Val           
80                  85                  90                  95            

ctg cat gtt gtt ggt gtt ccg agc att agc gca cag gca aaa cag ctg        336
Leu His Val Val Gly Val Pro Ser Ile Ser Ala Gln Ala Lys Gln Leu           
                100                 105                 110               

ctg ctg cat cat acc ctg ggt aat ggt gat ttt acc gtg ttt cat cgt        384
Leu Leu His His Thr Leu Gly Asn Gly Asp Phe Thr Val Phe His Arg           
            115                 120                 125                   

atg agc gca aat att agc gaa acc acc gca atg att acc gat att gca        432
Met Ser Ala Asn Ile Ser Glu Thr Thr Ala Met Ile Thr Asp Ile Ala           
        130                 135                 140                       

acc gca ccg gca gaa att gat cgt tgt att cgt acc acc tat gtt acc        480
Thr Ala Pro Ala Glu Ile Asp Arg Cys Ile Arg Thr Thr Tyr Val Thr           
    145                 150                 155                           

cag cgt ccg gtt tat ctg ggt ctg cca gca aat ctg gtt gat ctg aat        528
Gln Arg Pro Val Tyr Leu Gly Leu Pro Ala Asn Leu Val Asp Leu Asn           
160                 165                 170                 175           

gtt ccg gct aaa ctg ctg caa acc ccg att gat atg agc ctg aaa ccg        576
Val Pro Ala Lys Leu Leu Gln Thr Pro Ile Asp Met Ser Leu Lys Pro           
                180                 185                 190               

aat gat gca gaa agc gaa aaa gaa gtg att gat acc att ctg gcc ctg        624
Asn Asp Ala Glu Ser Glu Lys Glu Val Ile Asp Thr Ile Leu Ala Leu           
            195                 200                 205                   

gtt aaa gat gca aaa aat ccg gtt att ctg gca gat gcc tgt tgt agc        672
Val Lys Asp Ala Lys Asn Pro Val Ile Leu Ala Asp Ala Cys Cys Ser           
        210                 215                 220                       

cgt cat gat gtt aaa gca gaa acc aaa aaa ctg atc gac ctg acc cag        720
Arg His Asp Val Lys Ala Glu Thr Lys Lys Leu Ile Asp Leu Thr Gln           
    225                 230                 235                           

ttt ccg gca ttt gtt acc ccg atg ggt aaa ggt agc att gat gaa cag        768
Phe Pro Ala Phe Val Thr Pro Met Gly Lys Gly Ser Ile Asp Glu Gln           
240                 245                 250                 255           

cat ccg cgt tat ggt ggt gtt tat gtt ggc acc ctg agc aaa ccg gaa        816
His Pro Arg Tyr Gly Gly Val Tyr Val Gly Thr Leu Ser Lys Pro Glu           
                260                 265                 270               

gtt aaa gaa gca gtt gaa agc gca gat ctg att ctg agc gtt ggt gca        864
Val Lys Glu Ala Val Glu Ser Ala Asp Leu Ile Leu Ser Val Gly Ala           
            275                 280                 285                   

ctg ctg agt gat ttt aac acc ggt agc ttt tcg tat agc tac aaa acg        912
Leu Leu Ser Asp Phe Asn Thr Gly Ser Phe Ser Tyr Ser Tyr Lys Thr           
        290                 295                 300                       

aaa aac atc gtc gag ttt cat agc gat cac atg aaa att cgt aat gca        960
Lys Asn Ile Val Glu Phe His Ser Asp His Met Lys Ile Arg Asn Ala           
    305                 310                 315                           

acc ttt ccg ggt gtg cag atg aaa ttt gtt ctg caa aaa ctg ctg acc       1008
Thr Phe Pro Gly Val Gln Met Lys Phe Val Leu Gln Lys Leu Leu Thr           
320                 325                 330                 335           

acc att gca gat gca gca aaa ggt tat aaa ccg gtt gca gtt ccg gca       1056
Thr Ile Ala Asp Ala Ala Lys Gly Tyr Lys Pro Val Ala Val Pro Ala           
                340                 345                 350               

cgt aca ccg gca aat gca gcc gtt ccg gca tca aca ccg ctg aaa caa       1104
Arg Thr Pro Ala Asn Ala Ala Val Pro Ala Ser Thr Pro Leu Lys Gln           
            355                 360                 365                   

gaa tgg atg tgg aat cag ctg ggt aat ttt ctg caa gag ggt gat gtt       1152
Glu Trp Met Trp Asn Gln Leu Gly Asn Phe Leu Gln Glu Gly Asp Val           
        370                 375                 380                       

gtt att gca gaa acc ggc acc agc gca ttt ggt att aat cag acc acc       1200
Val Ile Ala Glu Thr Gly Thr Ser Ala Phe Gly Ile Asn Gln Thr Thr           
    385                 390                 395                           

ttt ccg aat aac acc tat ggt att agc cag gtt ctg tgg ggt agt att       1248
Phe Pro Asn Asn Thr Tyr Gly Ile Ser Gln Val Leu Trp Gly Ser Ile           
400                 405                 410                 415           

ggt ttt acc acc ggt gca acc ctg ggt gca gca ttt gca gcc gaa gaa       1296
Gly Phe Thr Thr Gly Ala Thr Leu Gly Ala Ala Phe Ala Ala Glu Glu           
                420                 425                 430               

atc gat ccg aaa aaa cgt gtg att ctg ttt att ggt gat ggt agc ctg       1344
Ile Asp Pro Lys Lys Arg Val Ile Leu Phe Ile Gly Asp Gly Ser Leu           
            435                 440                 445                   

caa ctg acc gtt caa gaa att agc acc atg att cgt tgg ggt ctg aaa       1392
Gln Leu Thr Val Gln Glu Ile Ser Thr Met Ile Arg Trp Gly Leu Lys           
        450                 455                 460                       

ccg tac ctg ttc gtt ctg aat aat gat ggc tat acc atc gag aaa ctg       1440
Pro Tyr Leu Phe Val Leu Asn Asn Asp Gly Tyr Thr Ile Glu Lys Leu           
    465                 470                 475                           

atc cac ggt ccg aaa gca cag tat aat gaa att cag ggt tgg gat cat       1488
Ile His Gly Pro Lys Ala Gln Tyr Asn Glu Ile Gln Gly Trp Asp His           
480                 485                 490                 495           

ctg tca ctg ctg ccg acc ttt ggc gca aaa gat tat gaa aca cat cgt       1536
Leu Ser Leu Leu Pro Thr Phe Gly Ala Lys Asp Tyr Glu Thr His Arg           
                500                 505                 510               

gtt gca acc aca ggt gaa tgg gat aaa ctg acc cag gat aaa tcc ttt       1584
Val Ala Thr Thr Gly Glu Trp Asp Lys Leu Thr Gln Asp Lys Ser Phe           
            515                 520                 525                   

aat gat aac agc aaa atc cgc atg atc gaa att atg ctg ccg gtt ttt       1632
Asn Asp Asn Ser Lys Ile Arg Met Ile Glu Ile Met Leu Pro Val Phe           
        530                 535                 540                       

gat gca ccg cag aat ctg gtg gaa cag gcc aaa ctg acc gca gca acc       1680
Asp Ala Pro Gln Asn Leu Val Glu Gln Ala Lys Leu Thr Ala Ala Thr           
    545                 550                 555                           

aat gca aaa cag ctc gag cac cac cac cac cac cac tga                   1719
Asn Ala Lys Gln Leu Glu His His His His His His                           
560                 565                 570                               


<210>  2
<211>  571
<212>  PRT
<213>  Saccharomyces cerevisiae

<400>  2

Met Ser Glu Ile Thr Leu Gly Lys Tyr Leu Phe Glu Arg Leu Lys Gln 
1               5                   10                  15      


Val Asn Val Asn Thr Val Phe Gly Leu Pro Gly Asp Phe Asn Leu Ser 
            20                  25                  30          


Leu Leu Asp Lys Ile Tyr Glu Val Glu Gly Met Arg Trp Ala Gly Asn 
        35                  40                  45              


Ala Asn Glu Leu Asn Ala Ala Tyr Ala Ala Asp Gly Tyr Ala Arg Ile 
    50                  55                  60                  


Lys Gly Met Ser Cys Ile Ile Thr Thr Phe Gly Val Gly Glu Leu Ser 
65                  70                  75                  80  


Ala Leu Asn Gly Ile Ala Gly Ser Tyr Ala Glu His Val Gly Val Leu 
                85                  90                  95      


His Val Val Gly Val Pro Ser Ile Ser Ala Gln Ala Lys Gln Leu Leu 
            100                 105                 110         


Leu His His Thr Leu Gly Asn Gly Asp Phe Thr Val Phe His Arg Met 
        115                 120                 125             


Ser Ala Asn Ile Ser Glu Thr Thr Ala Met Ile Thr Asp Ile Ala Thr 
    130                 135                 140                 


Ala Pro Ala Glu Ile Asp Arg Cys Ile Arg Thr Thr Tyr Val Thr Gln 
145                 150                 155                 160 


Arg Pro Val Tyr Leu Gly Leu Pro Ala Asn Leu Val Asp Leu Asn Val 
                165                 170                 175     


Pro Ala Lys Leu Leu Gln Thr Pro Ile Asp Met Ser Leu Lys Pro Asn 
            180                 185                 190         


Asp Ala Glu Ser Glu Lys Glu Val Ile Asp Thr Ile Leu Ala Leu Val 
        195                 200                 205             


Lys Asp Ala Lys Asn Pro Val Ile Leu Ala Asp Ala Cys Cys Ser Arg 
    210                 215                 220                 


His Asp Val Lys Ala Glu Thr Lys Lys Leu Ile Asp Leu Thr Gln Phe 
225                 230                 235                 240 


Pro Ala Phe Val Thr Pro Met Gly Lys Gly Ser Ile Asp Glu Gln His 
                245                 250                 255     


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


Lys Glu Ala Val Glu Ser Ala Asp Leu Ile Leu Ser Val Gly Ala Leu 
        275                 280                 285             


Leu Ser Asp Phe Asn Thr Gly Ser Phe Ser Tyr Ser Tyr Lys Thr Lys 
    290                 295                 300                 


Asn Ile Val Glu Phe His Ser Asp His Met Lys Ile Arg Asn Ala Thr 
305                 310                 315                 320 


Phe Pro Gly Val Gln Met Lys Phe Val Leu Gln Lys Leu Leu Thr Thr 
                325                 330                 335     


Ile Ala Asp Ala Ala Lys Gly Tyr Lys Pro Val Ala Val Pro Ala Arg 
            340                 345                 350         


Thr Pro Ala Asn Ala Ala Val Pro Ala Ser Thr Pro Leu Lys Gln Glu 
        355                 360                 365             


Trp Met Trp Asn Gln Leu Gly Asn Phe Leu Gln Glu Gly Asp Val Val 
    370                 375                 380                 


Ile Ala Glu Thr Gly Thr Ser Ala Phe Gly Ile Asn Gln Thr Thr Phe 
385                 390                 395                 400 


Pro Asn Asn Thr Tyr Gly Ile Ser Gln Val Leu Trp Gly Ser Ile Gly 
                405                 410                 415     


Phe Thr Thr Gly Ala Thr Leu Gly Ala Ala Phe Ala Ala Glu Glu Ile 
            420                 425                 430         


Asp Pro Lys Lys Arg Val Ile Leu Phe Ile Gly Asp Gly Ser Leu Gln 
        435                 440                 445             


Leu Thr Val Gln Glu Ile Ser Thr Met Ile Arg Trp Gly Leu Lys Pro 
    450                 455                 460                 


Tyr Leu Phe Val Leu Asn Asn Asp Gly Tyr Thr Ile Glu Lys Leu Ile 
465                 470                 475                 480 


His Gly Pro Lys Ala Gln Tyr Asn Glu Ile Gln Gly Trp Asp His Leu 
                485                 490                 495     


Ser Leu Leu Pro Thr Phe Gly Ala Lys Asp Tyr Glu Thr His Arg Val 
            500                 505                 510         


Ala Thr Thr Gly Glu Trp Asp Lys Leu Thr Gln Asp Lys Ser Phe Asn 
        515                 520                 525             


Asp Asn Ser Lys Ile Arg Met Ile Glu Ile Met Leu Pro Val Phe Asp 
    530                 535                 540                 


Ala Pro Gln Asn Leu Val Glu Gln Ala Lys Leu Thr Ala Ala Thr Asn 
545                 550                 555                 560 


Ala Lys Gln Leu Glu His His His His His His 
                565                 570     


<210>  3
<211>  1932
<212>  DNA
<213>  Saccharomyces cerevisiae


<220>
<221>  misc_feature
<222>  (1)..(6)
<223>  NdeI restriction site

<220>
<221>  CDS
<222>  (4)..(1932)
<223>  sequence of ARO10 of Sacharomyces cerevisiae optimized for the 
       codon usage of Escherichia coli with a carboxy-terminal His6-tag

<220>
<221>  misc_feature
<222>  (1906)..(1911)
<223>  XhoI restriction site

<400>  3
cat atg gca ccg gtt acc att gaa aaa ttc gtg aat caa gaa gaa cgc         48
    Met Ala Pro Val Thr Ile Glu Lys Phe Val Asn Gln Glu Glu Arg           
    1               5                   10                  15            

cac ctg gtt agc aat cgt agc gca acc att ccg ttt ggt gaa tat atc         96
His Leu Val Ser Asn Arg Ser Ala Thr Ile Pro Phe Gly Glu Tyr Ile           
                20                  25                  30                

ttt aaa cgc ctg ctg agc att gat acc aaa agc gtg ttt ggt gtt ccg        144
Phe Lys Arg Leu Leu Ser Ile Asp Thr Lys Ser Val Phe Gly Val Pro           
            35                  40                  45                    

ggt gat ttt aat ctg agc ctg ctg gaa tat ctg tat agc ccg agc gtt        192
Gly Asp Phe Asn Leu Ser Leu Leu Glu Tyr Leu Tyr Ser Pro Ser Val           
        50                  55                  60                        

gaa agc gca ggt ctg cgt tgg gtt ggc acc tgt aat gaa ctg aat gca        240
Glu Ser Ala Gly Leu Arg Trp Val Gly Thr Cys Asn Glu Leu Asn Ala           
    65                  70                  75                            

gcc tat gca gca gat ggt tat agc cgt tat agc aac aaa att ggt tgt        288
Ala Tyr Ala Ala Asp Gly Tyr Ser Arg Tyr Ser Asn Lys Ile Gly Cys           
80                  85                  90                  95            

ctg att acc acc tat ggt gtt ggt gaa ctg agc gca ctg aat ggt att        336
Leu Ile Thr Thr Tyr Gly Val Gly Glu Leu Ser Ala Leu Asn Gly Ile           
                100                 105                 110               

gca ggt agc ttt gca gaa aat gtg aaa gtg ctg cat att gtt ggt gtg        384
Ala Gly Ser Phe Ala Glu Asn Val Lys Val Leu His Ile Val Gly Val           
            115                 120                 125                   

gcc aaa agt att gat agc cgt agc agc aat ttt agc gat cgt aat ctg        432
Ala Lys Ser Ile Asp Ser Arg Ser Ser Asn Phe Ser Asp Arg Asn Leu           
        130                 135                 140                       

cat cat ctg gtt ccg cag ctg cat gat agc aac ttt aaa ggt ccg aac        480
His His Leu Val Pro Gln Leu His Asp Ser Asn Phe Lys Gly Pro Asn           
    145                 150                 155                           

cat aaa gtg tat cac gac atg gtt aaa gat cgt gtt gca tgt agc gtt        528
His Lys Val Tyr His Asp Met Val Lys Asp Arg Val Ala Cys Ser Val           
160                 165                 170                 175           

gca tat ctg gaa gat att gaa acc gca tgt gat cag gtg gat aat gtg        576
Ala Tyr Leu Glu Asp Ile Glu Thr Ala Cys Asp Gln Val Asp Asn Val           
                180                 185                 190               

att cgc gat atc tat aaa tac agc aaa ccg ggt tat atc ttt gtg cct        624
Ile Arg Asp Ile Tyr Lys Tyr Ser Lys Pro Gly Tyr Ile Phe Val Pro           
            195                 200                 205                   

gcc gat ttt gca gat atg agc gtt acc tgt gat aat ctg gtt aat gtt        672
Ala Asp Phe Ala Asp Met Ser Val Thr Cys Asp Asn Leu Val Asn Val           
        210                 215                 220                       

ccg cgt att agc cag cag gat tgt att gtt tat ccg agc gaa aat cag        720
Pro Arg Ile Ser Gln Gln Asp Cys Ile Val Tyr Pro Ser Glu Asn Gln           
    225                 230                 235                           

ctg agc gac att att aac aaa atc acc agc tgg atc tat agc agc aaa        768
Leu Ser Asp Ile Ile Asn Lys Ile Thr Ser Trp Ile Tyr Ser Ser Lys           
240                 245                 250                 255           

aca ccg gca att ctg ggt gat gtt ctg acc gat cgt tat ggt gtt agc        816
Thr Pro Ala Ile Leu Gly Asp Val Leu Thr Asp Arg Tyr Gly Val Ser           
                260                 265                 270               

aat ttt ctg aac aaa ctg att tgc aaa acc ggc atc tgg aat ttt tct        864
Asn Phe Leu Asn Lys Leu Ile Cys Lys Thr Gly Ile Trp Asn Phe Ser           
            275                 280                 285                   

acc gtt atg ggt aaa agc gtg atc gat gaa agc aat ccg acc tat atg        912
Thr Val Met Gly Lys Ser Val Ile Asp Glu Ser Asn Pro Thr Tyr Met           
        290                 295                 300                       

ggt cag tat aat ggt aaa gaa ggc ctg aaa cag gtc tat gaa cat ttt        960
Gly Gln Tyr Asn Gly Lys Glu Gly Leu Lys Gln Val Tyr Glu His Phe           
    305                 310                 315                           

gaa ctg tgt gat ctg gtg ctg cat ttt ggc gtt gac att aac gaa att       1008
Glu Leu Cys Asp Leu Val Leu His Phe Gly Val Asp Ile Asn Glu Ile           
320                 325                 330                 335           

aac aac ggc cat tac acc ttc acc tat aaa ccg aat gca aaa atc atc       1056
Asn Asn Gly His Tyr Thr Phe Thr Tyr Lys Pro Asn Ala Lys Ile Ile           
                340                 345                 350               

cag ttc cac ccg aac tat att cgt ctg gtt gat acc cgt cag ggt aat       1104
Gln Phe His Pro Asn Tyr Ile Arg Leu Val Asp Thr Arg Gln Gly Asn           
            355                 360                 365                   

gag cag atg ttt aaa ggt att aac ttt gca ccg atc ctg aaa gag ctg       1152
Glu Gln Met Phe Lys Gly Ile Asn Phe Ala Pro Ile Leu Lys Glu Leu           
        370                 375                 380                       

tat aaa cgt att gat gtg agc aaa ctg tcc ctg caa tat gat agt aat       1200
Tyr Lys Arg Ile Asp Val Ser Lys Leu Ser Leu Gln Tyr Asp Ser Asn           
    385                 390                 395                           

gtt acc cag tat acc aac gaa acc atg cgc ctg gaa gat ccg acc aat       1248
Val Thr Gln Tyr Thr Asn Glu Thr Met Arg Leu Glu Asp Pro Thr Asn           
400                 405                 410                 415           

ggt cag agc agc att att acc cag gtt cat ctg caa aaa acg atg ccg       1296
Gly Gln Ser Ser Ile Ile Thr Gln Val His Leu Gln Lys Thr Met Pro           
                420                 425                 430               

aaa ttt ctg aat cca ggt gat gtt gtt gtt tgt gaa acc ggt agc ttt       1344
Lys Phe Leu Asn Pro Gly Asp Val Val Val Cys Glu Thr Gly Ser Phe           
            435                 440                 445                   

cag ttt agc gtt cgt gat ttt gca ttt ccg agc cag ctg aaa tat atc       1392
Gln Phe Ser Val Arg Asp Phe Ala Phe Pro Ser Gln Leu Lys Tyr Ile           
        450                 455                 460                       

agc cag ggt ttt ttt ctg agt att ggt atg gca ctg cct gcc gca ctg       1440
Ser Gln Gly Phe Phe Leu Ser Ile Gly Met Ala Leu Pro Ala Ala Leu           
    465                 470                 475                           

ggt gtt ggt atc gca atg cag gat cat agc aat gca cat att aat ggt       1488
Gly Val Gly Ile Ala Met Gln Asp His Ser Asn Ala His Ile Asn Gly           
480                 485                 490                 495           

ggc aac gtg aaa gag gat tat aaa ccg cgt ctg att ctg ttt gaa ggt       1536
Gly Asn Val Lys Glu Asp Tyr Lys Pro Arg Leu Ile Leu Phe Glu Gly           
                500                 505                 510               

gat ggt gca gca cag atg acc att caa gaa ctg agc acc att ctg aaa       1584
Asp Gly Ala Ala Gln Met Thr Ile Gln Glu Leu Ser Thr Ile Leu Lys           
            515                 520                 525                   

tgt aat att ccg ctg gaa gtg atc atc tgg aac aat aat ggt tat acg       1632
Cys Asn Ile Pro Leu Glu Val Ile Ile Trp Asn Asn Asn Gly Tyr Thr           
        530                 535                 540                       

att gag cgt gca att atg ggt ccg acc cgt agc tat aat gat gtt atg       1680
Ile Glu Arg Ala Ile Met Gly Pro Thr Arg Ser Tyr Asn Asp Val Met           
    545                 550                 555                           

agc tgg aaa tgg acc aaa ctg ttt gag gca ttt ggc gat ttt gat ggc       1728
Ser Trp Lys Trp Thr Lys Leu Phe Glu Ala Phe Gly Asp Phe Asp Gly           
560                 565                 570                 575           

aaa tat acc aat agc acc ctg att cag tgt ccg agt aaa ctg gca ctg       1776
Lys Tyr Thr Asn Ser Thr Leu Ile Gln Cys Pro Ser Lys Leu Ala Leu           
                580                 585                 590               

aaa ctg gaa gaa ctg aaa aac agc aat aaa cgc agc ggt att gaa ctg       1824
Lys Leu Glu Glu Leu Lys Asn Ser Asn Lys Arg Ser Gly Ile Glu Leu           
            595                 600                 605                   

ctg gaa gtt aaa ctg ggc gaa ctg gat ttt ccg gaa caa ctg aaa tgc       1872
Leu Glu Val Lys Leu Gly Glu Leu Asp Phe Pro Glu Gln Leu Lys Cys           
        610                 615                 620                       

atg gtt gaa gca gca gcc ctg aaa cgt aat aaa ctc gag cac cac cac       1920
Met Val Glu Ala Ala Ala Leu Lys Arg Asn Lys Leu Glu His His His           
    625                 630                 635                           

cac cac cac tga                                                       1932
His His His                                                               
640                                                                       


<210>  4
<211>  642
<212>  PRT
<213>  Saccharomyces cerevisiae

<400>  4

Met Ala Pro Val Thr Ile Glu Lys Phe Val Asn Gln Glu Glu Arg His 
1               5                   10                  15      


Leu Val Ser Asn Arg Ser Ala Thr Ile Pro Phe Gly Glu Tyr Ile Phe 
            20                  25                  30          


Lys Arg Leu Leu Ser Ile Asp Thr Lys Ser Val Phe Gly Val Pro Gly 
        35                  40                  45              


Asp Phe Asn Leu Ser Leu Leu Glu Tyr Leu Tyr Ser Pro Ser Val Glu 
    50                  55                  60                  


Ser Ala Gly Leu Arg Trp Val Gly Thr Cys Asn Glu Leu Asn Ala Ala 
65                  70                  75                  80  


Tyr Ala Ala Asp Gly Tyr Ser Arg Tyr Ser Asn Lys Ile Gly Cys Leu 
                85                  90                  95      


Ile Thr Thr Tyr Gly Val Gly Glu Leu Ser Ala Leu Asn Gly Ile Ala 
            100                 105                 110         


Gly Ser Phe Ala Glu Asn Val Lys Val Leu His Ile Val Gly Val Ala 
        115                 120                 125             


Lys Ser Ile Asp Ser Arg Ser Ser Asn Phe Ser Asp Arg Asn Leu His 
    130                 135                 140                 


His Leu Val Pro Gln Leu His Asp Ser Asn Phe Lys Gly Pro Asn His 
145                 150                 155                 160 


Lys Val Tyr His Asp Met Val Lys Asp Arg Val Ala Cys Ser Val Ala 
                165                 170                 175     


Tyr Leu Glu Asp Ile Glu Thr Ala Cys Asp Gln Val Asp Asn Val Ile 
            180                 185                 190         


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


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


Arg Ile Ser Gln Gln Asp Cys Ile Val Tyr Pro Ser Glu Asn Gln Leu 
225                 230                 235                 240 


Ser Asp Ile Ile Asn Lys Ile Thr Ser Trp Ile Tyr Ser Ser Lys Thr 
                245                 250                 255     


Pro Ala Ile Leu Gly Asp Val Leu Thr Asp Arg Tyr Gly Val Ser Asn 
            260                 265                 270         


Phe Leu Asn Lys Leu Ile Cys Lys Thr Gly Ile Trp Asn Phe Ser Thr 
        275                 280                 285             


Val Met Gly Lys Ser Val Ile Asp Glu Ser Asn Pro Thr Tyr Met Gly 
    290                 295                 300                 


Gln Tyr Asn Gly Lys Glu Gly Leu Lys Gln Val Tyr Glu His Phe Glu 
305                 310                 315                 320 


Leu Cys Asp Leu Val Leu His Phe Gly Val Asp Ile Asn Glu Ile Asn 
                325                 330                 335     


Asn Gly His Tyr Thr Phe Thr Tyr Lys Pro Asn Ala Lys Ile Ile Gln 
            340                 345                 350         


Phe His Pro Asn Tyr Ile Arg Leu Val Asp Thr Arg Gln Gly Asn Glu 
        355                 360                 365             


Gln Met Phe Lys Gly Ile Asn Phe Ala Pro Ile Leu Lys Glu Leu Tyr 
    370                 375                 380                 


Lys Arg Ile Asp Val Ser Lys Leu Ser Leu Gln Tyr Asp Ser Asn Val 
385                 390                 395                 400 


Thr Gln Tyr Thr Asn Glu Thr Met Arg Leu Glu Asp Pro Thr Asn Gly 
                405                 410                 415     


Gln Ser Ser Ile Ile Thr Gln Val His Leu Gln Lys Thr Met Pro Lys 
            420                 425                 430         


Phe Leu Asn Pro Gly Asp Val Val Val Cys Glu Thr Gly Ser Phe Gln 
        435                 440                 445             


Phe Ser Val Arg Asp Phe Ala Phe Pro Ser Gln Leu Lys Tyr Ile Ser 
    450                 455                 460                 


Gln Gly Phe Phe Leu Ser Ile Gly Met Ala Leu Pro Ala Ala Leu Gly 
465                 470                 475                 480 


Val Gly Ile Ala Met Gln Asp His Ser Asn Ala His Ile Asn Gly Gly 
                485                 490                 495     


Asn Val Lys Glu Asp Tyr Lys Pro Arg Leu Ile Leu Phe Glu Gly Asp 
            500                 505                 510         


Gly Ala Ala Gln Met Thr Ile Gln Glu Leu Ser Thr Ile Leu Lys Cys 
        515                 520                 525             


Asn Ile Pro Leu Glu Val Ile Ile Trp Asn Asn Asn Gly Tyr Thr Ile 
    530                 535                 540                 


Glu Arg Ala Ile Met Gly Pro Thr Arg Ser Tyr Asn Asp Val Met Ser 
545                 550                 555                 560 


Trp Lys Trp Thr Lys Leu Phe Glu Ala Phe Gly Asp Phe Asp Gly Lys 
                565                 570                 575     


Tyr Thr Asn Ser Thr Leu Ile Gln Cys Pro Ser Lys Leu Ala Leu Lys 
            580                 585                 590         


Leu Glu Glu Leu Lys Asn Ser Asn Lys Arg Ser Gly Ile Glu Leu Leu 
        595                 600                 605             


Glu Val Lys Leu Gly Glu Leu Asp Phe Pro Glu Gln Leu Lys Cys Met 
    610                 615                 620                 


Val Glu Ala Ala Ala Leu Lys Arg Asn Lys Leu Glu His His His His 
625                 630                 635                 640 


His His 
        


<210>  5
<211>  1671
<212>  DNA
<213>  Lactococcus lactis


<220>
<221>  misc_feature
<222>  (1)..(6)
<223>  NdeI restriction site

<220>
<221>  CDS
<222>  (4)..(1671)
<223>  sequence of kdcA of Lactococcus lactis optimized for the codon 
       usage of Escherichia coli with a carboxy-terminal His6-tag

<220>
<221>  misc_feature
<222>  (1645)..(1650)
<223>  XhoI restriction site

<400>  5
cat atg tat acc gtt ggt gat tat ctg ctg gat cgt ctg cat gaa ctg         48
    Met Tyr Thr Val Gly Asp Tyr Leu Leu Asp Arg Leu His Glu Leu           
    1               5                   10                  15            

ggt att gaa gaa att ttt ggt gtt ccg ggt gat tac aac ctg caa ttt         96
Gly Ile Glu Glu Ile Phe Gly Val Pro Gly Asp Tyr Asn Leu Gln Phe           
                20                  25                  30                

ctg gat cag att atc agc cgt gaa gat atg aaa tgg att ggc aat gcg        144
Leu Asp Gln Ile Ile Ser Arg Glu Asp Met Lys Trp Ile Gly Asn Ala           
            35                  40                  45                    

aat gaa ctg aat gca agc tat atg gca gat ggt tat gca cgt acc aaa        192
Asn Glu Leu Asn Ala Ser Tyr Met Ala Asp Gly Tyr Ala Arg Thr Lys           
        50                  55                  60                        

aaa gca gca gca ttt ctg acc acc ttt ggt gtt ggt gaa ctg agc gca        240
Lys Ala Ala Ala Phe Leu Thr Thr Phe Gly Val Gly Glu Leu Ser Ala           
    65                  70                  75                            

att aat ggt ctg gca ggt agc tat gca gaa aat ctg ccg gtt gtt gaa        288
Ile Asn Gly Leu Ala Gly Ser Tyr Ala Glu Asn Leu Pro Val Val Glu           
80                  85                  90                  95            

att gtt ggt agc ccg acc agc aaa gtt cag aat gat ggt aaa ttt gtg        336
Ile Val Gly Ser Pro Thr Ser Lys Val Gln Asn Asp Gly Lys Phe Val           
                100                 105                 110               

cat cat acc ctg gcc gat ggt gat ttt aaa cac ttt atg aaa atg cac        384
His His Thr Leu Ala Asp Gly Asp Phe Lys His Phe Met Lys Met His           
            115                 120                 125                   

gaa ccg gtt acc gca gca cgt acc ctg ctg acc gca gaa aat gca acc        432
Glu Pro Val Thr Ala Ala Arg Thr Leu Leu Thr Ala Glu Asn Ala Thr           
        130                 135                 140                       

tat gaa att gat cgt gtt ctg agc cag ctg ctg aaa gaa cgt aaa ccg        480
Tyr Glu Ile Asp Arg Val Leu Ser Gln Leu Leu Lys Glu Arg Lys Pro           
    145                 150                 155                           

gtg tat att aac ctg ccg gtg gat gtt gca gca gca aaa gca gaa aaa        528
Val Tyr Ile Asn Leu Pro Val Asp Val Ala Ala Ala Lys Ala Glu Lys           
160                 165                 170                 175           

ccg gca ctg agc ctg gaa aaa gaa agc agc acc acc aat acc acc gaa        576
Pro Ala Leu Ser Leu Glu Lys Glu Ser Ser Thr Thr Asn Thr Thr Glu           
                180                 185                 190               

cag gtt att ctg agc aaa atc gaa gaa agc ctg aaa aat gcc cag aaa        624
Gln Val Ile Leu Ser Lys Ile Glu Glu Ser Leu Lys Asn Ala Gln Lys           
            195                 200                 205                   

ccg gtt gtt att gca ggt cat gaa gtt att agc ttt ggg ctg gaa aaa        672
Pro Val Val Ile Ala Gly His Glu Val Ile Ser Phe Gly Leu Glu Lys           
        210                 215                 220                       

acc gtt acc cag ttt gtt agc gaa acc aaa ctg ccg att acc acc ctg        720
Thr Val Thr Gln Phe Val Ser Glu Thr Lys Leu Pro Ile Thr Thr Leu           
    225                 230                 235                           

aat ttt ggt aaa agc gca gtt gat gaa agc ctg ccg agc ttt ctg ggt        768
Asn Phe Gly Lys Ser Ala Val Asp Glu Ser Leu Pro Ser Phe Leu Gly           
240                 245                 250                 255           

atc tat aat ggt aaa ctg tcc gag atc tcc ctg aaa aac ttt gtt gaa        816
Ile Tyr Asn Gly Lys Leu Ser Glu Ile Ser Leu Lys Asn Phe Val Glu           
                260                 265                 270               

agc gca gat ttc att ctg atg ctg ggt gtt aaa ctg acc gat agc agt        864
Ser Ala Asp Phe Ile Leu Met Leu Gly Val Lys Leu Thr Asp Ser Ser           
            275                 280                 285                   

acc ggt gca ttt acc cat cat ctg gat gaa aac aaa atg atc agc ctg        912
Thr Gly Ala Phe Thr His His Leu Asp Glu Asn Lys Met Ile Ser Leu           
        290                 295                 300                       

aac att gat gag ggc atc atc ttt aac aaa gtg gtg gaa gat ttt gat        960
Asn Ile Asp Glu Gly Ile Ile Phe Asn Lys Val Val Glu Asp Phe Asp           
    305                 310                 315                           

ttt cgt gca gtt gtt agc agc ctg agc gaa ctg aaa ggt att gaa tat       1008
Phe Arg Ala Val Val Ser Ser Leu Ser Glu Leu Lys Gly Ile Glu Tyr           
320                 325                 330                 335           

gaa ggc cag tac atc gac aaa cag tat gaa gaa ttt att ccg agc agc       1056
Glu Gly Gln Tyr Ile Asp Lys Gln Tyr Glu Glu Phe Ile Pro Ser Ser           
                340                 345                 350               

gca ccg ctg agc cag gat cgc ctg tgg cag gca gtt gaa agt ctg acc       1104
Ala Pro Leu Ser Gln Asp Arg Leu Trp Gln Ala Val Glu Ser Leu Thr           
            355                 360                 365                   

cag agc aat gaa acc att gtt gca gaa cag ggc acc agt ttt ttt ggt       1152
Gln Ser Asn Glu Thr Ile Val Ala Glu Gln Gly Thr Ser Phe Phe Gly           
        370                 375                 380                       

gca agc acc att ttt ctg aaa agc aac agc cgt ttt att ggt cag ccg       1200
Ala Ser Thr Ile Phe Leu Lys Ser Asn Ser Arg Phe Ile Gly Gln Pro           
    385                 390                 395                           

ctg tgg ggt agc att ggt tat acc ttt ccg gca gca ctg ggt agc cag       1248
Leu Trp Gly Ser Ile Gly Tyr Thr Phe Pro Ala Ala Leu Gly Ser Gln           
400                 405                 410                 415           

att gca gat aaa gaa agc cgt cat ctg ctg ttt att ggt gat ggt agc       1296
Ile Ala Asp Lys Glu Ser Arg His Leu Leu Phe Ile Gly Asp Gly Ser           
                420                 425                 430               

ctg caa ctg acc gtt caa gaa ctg ggt ctg agc att cgt gaa aaa ctg       1344
Leu Gln Leu Thr Val Gln Glu Leu Gly Leu Ser Ile Arg Glu Lys Leu           
            435                 440                 445                   

aat ccg att tgc ttc atc att aac aac gat ggc tat acc gtg gaa cgt       1392
Asn Pro Ile Cys Phe Ile Ile Asn Asn Asp Gly Tyr Thr Val Glu Arg           
        450                 455                 460                       

gaa att cat ggt ccg acc cag agt tat aat gat att ccg atg tgg aac       1440
Glu Ile His Gly Pro Thr Gln Ser Tyr Asn Asp Ile Pro Met Trp Asn           
    465                 470                 475                           

tac tcg aaa ctg cct gaa acc ttt ggc gca acc gaa gat cgt gtg gtt       1488
Tyr Ser Lys Leu Pro Glu Thr Phe Gly Ala Thr Glu Asp Arg Val Val           
480                 485                 490                 495           

agc aaa att gtt cgt acc gaa aat gaa ttt gtg agc gtg atg aaa gaa       1536
Ser Lys Ile Val Arg Thr Glu Asn Glu Phe Val Ser Val Met Lys Glu           
                500                 505                 510               

gca cag gca gac gtt aat cgt atg tat tgg att gaa ctg gtg ctg gaa       1584
Ala Gln Ala Asp Val Asn Arg Met Tyr Trp Ile Glu Leu Val Leu Glu           
            515                 520                 525                   

aaa gag gat gca ccg aaa ctg ctg aaa aaa atg ggt aaa ctg ttt gcc       1632
Lys Glu Asp Ala Pro Lys Leu Leu Lys Lys Met Gly Lys Leu Phe Ala           
        530                 535                 540                       

gag cag aat aaa ctc gag cac cac cac cac cac cac tga                   1671
Glu Gln Asn Lys Leu Glu His His His His His His                           
    545                 550                 555                           


<210>  6
<211>  555
<212>  PRT
<213>  Lactococcus lactis

<400>  6

Met Tyr Thr Val Gly Asp Tyr Leu Leu Asp Arg Leu His Glu Leu Gly 
1               5                   10                  15      


Ile Glu Glu Ile Phe Gly Val Pro Gly Asp Tyr Asn Leu Gln Phe Leu 
            20                  25                  30          


Asp Gln Ile Ile Ser Arg Glu Asp Met Lys Trp Ile Gly Asn Ala Asn 
        35                  40                  45              


Glu Leu Asn Ala Ser Tyr Met Ala Asp Gly Tyr Ala Arg Thr Lys Lys 
    50                  55                  60                  


Ala Ala Ala Phe Leu Thr Thr Phe Gly Val Gly Glu Leu Ser Ala Ile 
65                  70                  75                  80  


Asn Gly Leu Ala Gly Ser Tyr Ala Glu Asn Leu Pro Val Val Glu Ile 
                85                  90                  95      


Val Gly Ser Pro Thr Ser Lys Val Gln Asn Asp Gly Lys Phe Val His 
            100                 105                 110         


His Thr Leu Ala Asp Gly Asp Phe Lys His Phe Met Lys Met His Glu 
        115                 120                 125             


Pro Val Thr Ala Ala Arg Thr Leu Leu Thr Ala Glu Asn Ala Thr Tyr 
    130                 135                 140                 


Glu Ile Asp Arg Val Leu Ser Gln Leu Leu Lys Glu Arg Lys Pro Val 
145                 150                 155                 160 


Tyr Ile Asn Leu Pro Val Asp Val Ala Ala Ala Lys Ala Glu Lys Pro 
                165                 170                 175     


Ala Leu Ser Leu Glu Lys Glu Ser Ser Thr Thr Asn Thr Thr Glu Gln 
            180                 185                 190         


Val Ile Leu Ser Lys Ile Glu Glu Ser Leu Lys Asn Ala Gln Lys Pro 
        195                 200                 205             


Val Val Ile Ala Gly His Glu Val Ile Ser Phe Gly Leu Glu Lys Thr 
    210                 215                 220                 


Val Thr Gln Phe Val Ser Glu Thr Lys Leu Pro Ile Thr Thr Leu Asn 
225                 230                 235                 240 


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


Tyr Asn Gly Lys Leu Ser Glu Ile Ser Leu Lys Asn Phe Val Glu Ser 
            260                 265                 270         


Ala Asp Phe Ile Leu Met Leu Gly Val Lys Leu Thr Asp Ser Ser Thr 
        275                 280                 285             


Gly Ala Phe Thr His His Leu Asp Glu Asn Lys Met Ile Ser Leu Asn 
    290                 295                 300                 


Ile Asp Glu Gly Ile Ile Phe Asn Lys Val Val Glu Asp Phe Asp Phe 
305                 310                 315                 320 


Arg Ala Val Val Ser Ser Leu Ser Glu Leu Lys Gly Ile Glu Tyr Glu 
                325                 330                 335     


Gly Gln Tyr Ile Asp Lys Gln Tyr Glu Glu Phe Ile Pro Ser Ser Ala 
            340                 345                 350         


Pro Leu Ser Gln Asp Arg Leu Trp Gln Ala Val Glu Ser Leu Thr Gln 
        355                 360                 365             


Ser Asn Glu Thr Ile Val Ala Glu Gln Gly Thr Ser Phe Phe Gly Ala 
    370                 375                 380                 


Ser Thr Ile Phe Leu Lys Ser Asn Ser Arg Phe Ile Gly Gln Pro Leu 
385                 390                 395                 400 


Trp Gly Ser Ile Gly Tyr Thr Phe Pro Ala Ala Leu Gly Ser Gln Ile 
                405                 410                 415     


Ala Asp Lys Glu Ser Arg His Leu Leu Phe Ile Gly Asp Gly Ser Leu 
            420                 425                 430         


Gln Leu Thr Val Gln Glu Leu Gly Leu Ser Ile Arg Glu Lys Leu Asn 
        435                 440                 445             


Pro Ile Cys Phe Ile Ile Asn Asn Asp Gly Tyr Thr Val Glu Arg Glu 
    450                 455                 460                 


Ile His Gly Pro Thr Gln Ser Tyr Asn Asp Ile Pro Met Trp Asn Tyr 
465                 470                 475                 480 


Ser Lys Leu Pro Glu Thr Phe Gly Ala Thr Glu Asp Arg Val Val Ser 
                485                 490                 495     


Lys Ile Val Arg Thr Glu Asn Glu Phe Val Ser Val Met Lys Glu Ala 
            500                 505                 510         


Gln Ala Asp Val Asn Arg Met Tyr Trp Ile Glu Leu Val Leu Glu Lys 
        515                 520                 525             


Glu Asp Ala Pro Lys Leu Leu Lys Lys Met Gly Lys Leu Phe Ala Glu 
    530                 535                 540                 


Gln Asn Lys Leu Glu His His His His His His 
545                 550                 555 


<210>  7
<211>  1207
<212>  DNA
<213>  Escherichia coli


<220>
<221>  CDS
<222>  (1)..(1203)
<223>  sequence of ybdL of Escherichia coli with an amino-terminal 
       His6-tag

<220>
<221>  misc_feature
<222>  (37)..(42)
<223>  KasI restriction site

<220>
<221>  misc_feature
<222>  (1202)..(1207)
<223>  HindIII restriction site

<400>  7
atg gct agc aga gga tcg cat cac cat cac cat cac ggc gcc atg aca         48
Met Ala Ser Arg Gly Ser His His His His His His Gly Ala Met Thr           
1               5                   10                  15                

aat aac cct ctg att cca caa agc aaa ctt cca caa ctt ggc acc act         96
Asn Asn Pro Leu Ile Pro Gln Ser Lys Leu Pro Gln Leu Gly Thr Thr           
            20                  25                  30                    

att ttc acc cag atg agc gcg ctg gcg cag caa cac cag gcg att aac        144
Ile Phe Thr Gln Met Ser Ala Leu Ala Gln Gln His Gln Ala Ile Asn           
        35                  40                  45                        

ctg tcg caa ggc ttt cct gat ttt gat ggt ccg cgc tat tta cag gag        192
Leu Ser Gln Gly Phe Pro Asp Phe Asp Gly Pro Arg Tyr Leu Gln Glu           
    50                  55                  60                            

cgg ctg gcg cac cac gtt gca cag ggg gca aac caa tac gcg ccc atg        240
Arg Leu Ala His His Val Ala Gln Gly Ala Asn Gln Tyr Ala Pro Met           
65                  70                  75                  80            

acc ggc gtg cag gcc ttg cgc gag gcg att gct cag aaa acg gaa cgt        288
Thr Gly Val Gln Ala Leu Arg Glu Ala Ile Ala Gln Lys Thr Glu Arg           
                85                  90                  95                

ttg tat ggc tat caa cca gat gcc gat agc gat atc acc gta acg gca        336
Leu Tyr Gly Tyr Gln Pro Asp Ala Asp Ser Asp Ile Thr Val Thr Ala           
            100                 105                 110                   

ggg gcg acg gaa gcg tta tac gcg gcg att acc gca ctg gtg cgc aat        384
Gly Ala Thr Glu Ala Leu Tyr Ala Ala Ile Thr Ala Leu Val Arg Asn           
        115                 120                 125                       

ggc gat gaa gtg att tgt ttt gat ccc agc tat gac agt tac gcc ccc        432
Gly Asp Glu Val Ile Cys Phe Asp Pro Ser Tyr Asp Ser Tyr Ala Pro           
    130                 135                 140                           

gcc atc gcg ctt tct ggg gga ata gtg aag cgt atg gca ctg caa cca        480
Ala Ile Ala Leu Ser Gly Gly Ile Val Lys Arg Met Ala Leu Gln Pro           
145                 150                 155                 160           

ccg cat ttt cgc gtt gac tgg cag gaa ttt gcc gca tta tta agc gag        528
Pro His Phe Arg Val Asp Trp Gln Glu Phe Ala Ala Leu Leu Ser Glu           
                165                 170                 175               

cgc acc aga ctg gtg atc ctc aac act ccg cat aac ccc agt gca act        576
Arg Thr Arg Leu Val Ile Leu Asn Thr Pro His Asn Pro Ser Ala Thr           
            180                 185                 190                   

gtc tgg cag cag gct gat ttc gcc gct ttg tgg cag gcg atc gcc ggg        624
Val Trp Gln Gln Ala Asp Phe Ala Ala Leu Trp Gln Ala Ile Ala Gly           
        195                 200                 205                       

cac gag att ttt gtc att agc gat gaa gtc tac gag cac atc aac ttt        672
His Glu Ile Phe Val Ile Ser Asp Glu Val Tyr Glu His Ile Asn Phe           
    210                 215                 220                           

tca caa cag ggc cat gcc agt gtg ctg gcg cat ccg cag ctg cgt gag        720
Ser Gln Gln Gly His Ala Ser Val Leu Ala His Pro Gln Leu Arg Glu           
225                 230                 235                 240           

cgg gca gtg gcg gtt tct tca ttt ggc aag acc tat cat atg acc ggc        768
Arg Ala Val Ala Val Ser Ser Phe Gly Lys Thr Tyr His Met Thr Gly           
                245                 250                 255               

tgg aaa gtg ggt tat tgt gtt gcg cca gcg ccc atc agc gcc gaa att        816
Trp Lys Val Gly Tyr Cys Val Ala Pro Ala Pro Ile Ser Ala Glu Ile           
            260                 265                 270                   

cgc aag gta cat cag tat ctg acc ttt tcg gtg aat acc ccg gca cag        864
Arg Lys Val His Gln Tyr Leu Thr Phe Ser Val Asn Thr Pro Ala Gln           
        275                 280                 285                       

ctg gcg ctt gct gat atg cta cgt gca gaa cct gag cat tat ctt gcg        912
Leu Ala Leu Ala Asp Met Leu Arg Ala Glu Pro Glu His Tyr Leu Ala           
    290                 295                 300                           

tta ccg gac ttt tat cgc cag aag cgc gat att ctg gtg aat gct tta        960
Leu Pro Asp Phe Tyr Arg Gln Lys Arg Asp Ile Leu Val Asn Ala Leu           
305                 310                 315                 320           

aat gaa agc cgg ctg gag att tta ccg tgt gaa ggt aca tac ttt ttg       1008
Asn Glu Ser Arg Leu Glu Ile Leu Pro Cys Glu Gly Thr Tyr Phe Leu           
                325                 330                 335               

ctg gtg gat tac agc gcg gtt tct acc ctg gat gat gtt gag ttt tgc       1056
Leu Val Asp Tyr Ser Ala Val Ser Thr Leu Asp Asp Val Glu Phe Cys           
            340                 345                 350                   

cag tgg ctg acg cag gag cac ggc gta gcg gcg att ccg ctg tcg gtg       1104
Gln Trp Leu Thr Gln Glu His Gly Val Ala Ala Ile Pro Leu Ser Val           
        355                 360                 365                       

ttt tgc gcc gat ccc ttc cca cat aaa ctg att cgt ctc tgt ttt gcc       1152
Phe Cys Ala Asp Pro Phe Pro His Lys Leu Ile Arg Leu Cys Phe Ala           
    370                 375                 380                           

aag aag gaa tcg acg ttg ctg gca gca gct gaa cgc ctg cgc cag ctt       1200
Lys Lys Glu Ser Thr Leu Leu Ala Ala Ala Glu Arg Leu Arg Gln Leu           
385                 390                 395                 400           

taa gctt                                                              1207


<210>  8
<211>  400
<212>  PRT
<213>  Escherichia coli

<400>  8

Met Ala Ser Arg Gly Ser His His His His His His Gly Ala Met Thr 
1               5                   10                  15      


Asn Asn Pro Leu Ile Pro Gln Ser Lys Leu Pro Gln Leu Gly Thr Thr 
            20                  25                  30          


Ile Phe Thr Gln Met Ser Ala Leu Ala Gln Gln His Gln Ala Ile Asn 
        35                  40                  45              


Leu Ser Gln Gly Phe Pro Asp Phe Asp Gly Pro Arg Tyr Leu Gln Glu 
    50                  55                  60                  


Arg Leu Ala His His Val Ala Gln Gly Ala Asn Gln Tyr Ala Pro Met 
65                  70                  75                  80  


Thr Gly Val Gln Ala Leu Arg Glu Ala Ile Ala Gln Lys Thr Glu Arg 
                85                  90                  95      


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


Gly Ala Thr Glu Ala Leu Tyr Ala Ala Ile Thr Ala Leu Val Arg Asn 
        115                 120                 125             


Gly Asp Glu Val Ile Cys Phe Asp Pro Ser Tyr Asp Ser Tyr Ala Pro 
    130                 135                 140                 


Ala Ile Ala Leu Ser Gly Gly Ile Val Lys Arg Met Ala Leu Gln Pro 
145                 150                 155                 160 


Pro His Phe Arg Val Asp Trp Gln Glu Phe Ala Ala Leu Leu Ser Glu 
                165                 170                 175     


Arg Thr Arg Leu Val Ile Leu Asn Thr Pro His Asn Pro Ser Ala Thr 
            180                 185                 190         


Val Trp Gln Gln Ala Asp Phe Ala Ala Leu Trp Gln Ala Ile Ala Gly 
        195                 200                 205             


His Glu Ile Phe Val Ile Ser Asp Glu Val Tyr Glu His Ile Asn Phe 
    210                 215                 220                 


Ser Gln Gln Gly His Ala Ser Val Leu Ala His Pro Gln Leu Arg Glu 
225                 230                 235                 240 


Arg Ala Val Ala Val Ser Ser Phe Gly Lys Thr Tyr His Met Thr Gly 
                245                 250                 255     


Trp Lys Val Gly Tyr Cys Val Ala Pro Ala Pro Ile Ser Ala Glu Ile 
            260                 265                 270         


Arg Lys Val His Gln Tyr Leu Thr Phe Ser Val Asn Thr Pro Ala Gln 
        275                 280                 285             


Leu Ala Leu Ala Asp Met Leu Arg Ala Glu Pro Glu His Tyr Leu Ala 
    290                 295                 300                 


Leu Pro Asp Phe Tyr Arg Gln Lys Arg Asp Ile Leu Val Asn Ala Leu 
305                 310                 315                 320 


Asn Glu Ser Arg Leu Glu Ile Leu Pro Cys Glu Gly Thr Tyr Phe Leu 
                325                 330                 335     


Leu Val Asp Tyr Ser Ala Val Ser Thr Leu Asp Asp Val Glu Phe Cys 
            340                 345                 350         


Gln Trp Leu Thr Gln Glu His Gly Val Ala Ala Ile Pro Leu Ser Val 
        355                 360                 365             


Phe Cys Ala Asp Pro Phe Pro His Lys Leu Ile Arg Leu Cys Phe Ala 
    370                 375                 380                 


Lys Lys Glu Ser Thr Leu Leu Ala Ala Ala Glu Arg Leu Arg Gln Leu 
385                 390                 395                 400 


<210>  9
<211>  1549
<212>  DNA
<213>  Saccharomyces cerevisiae


<220>
<221>  CDS
<222>  (1)..(1545)
<223>  sequence of ARO8 of Saccharomyces cerevisiae optimized for the 
       codon usage of Escherichia coli with an amino-terminal His6-tag

<220>
<221>  misc_feature
<222>  (37)..(42)
<223>  KasI restriction site

<220>
<221>  misc_feature
<222>  (1544)..(1549)
<223>  HindIII restriction site

<400>  9
atg gct agc aga gga tcg cat cac cat cac cat cac ggc gcc atg acc         48
Met Ala Ser Arg Gly Ser His His His His His His Gly Ala Met Thr           
1               5                   10                  15                

ctg ccg gaa agc aaa gat ttt agc tac ctg ttt tcc gat gaa acc aat         96
Leu Pro Glu Ser Lys Asp Phe Ser Tyr Leu Phe Ser Asp Glu Thr Asn           
            20                  25                  30                    

gca cgt aaa ccg agt ccg ctg aaa acc tgt att cat tta ttt cag gat        144
Ala Arg Lys Pro Ser Pro Leu Lys Thr Cys Ile His Leu Phe Gln Asp           
        35                  40                  45                        

ccg aac atc atc ttt ctg ggt ggt ggt ctg ccg ctg aaa gat tat ttt        192
Pro Asn Ile Ile Phe Leu Gly Gly Gly Leu Pro Leu Lys Asp Tyr Phe           
    50                  55                  60                            

ccg tgg gat aat ctg agc gtt gat agc ccg aaa ccg cct ttt ccg cag        240
Pro Trp Asp Asn Leu Ser Val Asp Ser Pro Lys Pro Pro Phe Pro Gln           
65                  70                  75                  80            

ggt att ggt gca ccg att gat gaa cag aat tgc atc aaa tac acc gtg        288
Gly Ile Gly Ala Pro Ile Asp Glu Gln Asn Cys Ile Lys Tyr Thr Val           
                85                  90                  95                

aac aaa gat tac gca gat aaa agc gca aat ccg agc aat gat att ccg        336
Asn Lys Asp Tyr Ala Asp Lys Ser Ala Asn Pro Ser Asn Asp Ile Pro           
            100                 105                 110                   

ctg agc cgt gca ctg cag tat ggt ttt agc gca ggt cag ccg gaa ctg        384
Leu Ser Arg Ala Leu Gln Tyr Gly Phe Ser Ala Gly Gln Pro Glu Leu           
        115                 120                 125                       

ctg aac ttt att cgt gat cat acc aaa atc atc cac gat ctg aaa tac        432
Leu Asn Phe Ile Arg Asp His Thr Lys Ile Ile His Asp Leu Lys Tyr           
    130                 135                 140                           

aaa gat tgg gat gtt ctg gca acc gca ggt aat acc aat gca tgg gaa        480
Lys Asp Trp Asp Val Leu Ala Thr Ala Gly Asn Thr Asn Ala Trp Glu           
145                 150                 155                 160           

agc acc ctg cgt gtt ttt tgt aat cgt ggt gat gtt att ctg gtt gaa        528
Ser Thr Leu Arg Val Phe Cys Asn Arg Gly Asp Val Ile Leu Val Glu           
                165                 170                 175               

gca cat agc ttt agc agc agc ctg gca agt gcc gaa gca cag ggt gtt        576
Ala His Ser Phe Ser Ser Ser Leu Ala Ser Ala Glu Ala Gln Gly Val           
            180                 185                 190                   

att acc ttt ccg gtt ccg atc gat gca gat ggt att att ccg gaa aaa        624
Ile Thr Phe Pro Val Pro Ile Asp Ala Asp Gly Ile Ile Pro Glu Lys           
        195                 200                 205                       

ctg gcc aaa gtg atg gaa aat tgg aca ccg ggt gct ccg aaa ccg aaa        672
Leu Ala Lys Val Met Glu Asn Trp Thr Pro Gly Ala Pro Lys Pro Lys           
    210                 215                 220                           

ctg ctg tat acc att ccg aca ggt cag aat ccg acc ggc acc agc att        720
Leu Leu Tyr Thr Ile Pro Thr Gly Gln Asn Pro Thr Gly Thr Ser Ile           
225                 230                 235                 240           

gca gat cat cgt aaa gaa gcc atc tat aaa atc gcc cag aaa tac gat        768
Ala Asp His Arg Lys Glu Ala Ile Tyr Lys Ile Ala Gln Lys Tyr Asp           
                245                 250                 255               

ttc ctg atc gtt gaa gat gag ccg tat tat ttc ctg cag atg aac ccg        816
Phe Leu Ile Val Glu Asp Glu Pro Tyr Tyr Phe Leu Gln Met Asn Pro           
            260                 265                 270                   

tat atc aaa gat ctg aaa gaa cgt gaa aaa gca cag agc agt ccg aaa        864
Tyr Ile Lys Asp Leu Lys Glu Arg Glu Lys Ala Gln Ser Ser Pro Lys           
        275                 280                 285                       

cag gat cat gat gaa ttt ctg aaa agc ctg gcc aat acc ttt ctg agc        912
Gln Asp His Asp Glu Phe Leu Lys Ser Leu Ala Asn Thr Phe Leu Ser           
    290                 295                 300                           

ctg gat acc gaa ggt cgt gtt att cgt atg gat agc ttt tca aaa gtt        960
Leu Asp Thr Glu Gly Arg Val Ile Arg Met Asp Ser Phe Ser Lys Val           
305                 310                 315                 320           

ctg gca ccg ggt aca cgt ctg ggt tgg att acc ggt agc agc aaa att       1008
Leu Ala Pro Gly Thr Arg Leu Gly Trp Ile Thr Gly Ser Ser Lys Ile           
                325                 330                 335               

ctg aaa ccg tat ctg agt ctg cat gaa atg acc att cag gca ccg gca       1056
Leu Lys Pro Tyr Leu Ser Leu His Glu Met Thr Ile Gln Ala Pro Ala           
            340                 345                 350                   

ggt ttt acc cag gtt ctg gtt aat gca acc ctg agc cgt tgg ggt cag       1104
Gly Phe Thr Gln Val Leu Val Asn Ala Thr Leu Ser Arg Trp Gly Gln           
        355                 360                 365                       

aaa ggc tat ctg gat tgg ctg ctg ggt ctg cgt cat gaa tat acc ctg       1152
Lys Gly Tyr Leu Asp Trp Leu Leu Gly Leu Arg His Glu Tyr Thr Leu           
    370                 375                 380                           

aaa cgt gat tgt gca att gat gcc ctg tat aaa tac ctg ccg cag agt       1200
Lys Arg Asp Cys Ala Ile Asp Ala Leu Tyr Lys Tyr Leu Pro Gln Ser           
385                 390                 395                 400           

gat gca ttt gtt att aat ccg cct att gcc ggt atg ttc ttt acc gtt       1248
Asp Ala Phe Val Ile Asn Pro Pro Ile Ala Gly Met Phe Phe Thr Val           
                405                 410                 415               

aat att gat gca agc gtg cac ccg gaa ttc aaa acc aaa tat aac agc       1296
Asn Ile Asp Ala Ser Val His Pro Glu Phe Lys Thr Lys Tyr Asn Ser           
            420                 425                 430                   

gat ccg tat cag ctg gaa cag agc ctg tat cat aaa gtt gtt gaa cgt       1344
Asp Pro Tyr Gln Leu Glu Gln Ser Leu Tyr His Lys Val Val Glu Arg           
        435                 440                 445                       

ggt gtt ctg gtt gtt ccg ggt agc tgg ttt aaa agc gaa ggt gaa acc       1392
Gly Val Leu Val Val Pro Gly Ser Trp Phe Lys Ser Glu Gly Glu Thr           
    450                 455                 460                           

gaa ccg cct cag cct gca gaa agt aaa gaa gtt agc aat ccg aat att       1440
Glu Pro Pro Gln Pro Ala Glu Ser Lys Glu Val Ser Asn Pro Asn Ile           
465                 470                 475                 480           

atc ttc ttc cgt ggc acc tat gca gca gtg agc cct gaa aaa ctg acc       1488
Ile Phe Phe Arg Gly Thr Tyr Ala Ala Val Ser Pro Glu Lys Leu Thr           
                485                 490                 495               

gaa ggc ctg aaa cgc ctg ggt gat acc ctg tat gaa gaa ttt ggt atc       1536
Glu Gly Leu Lys Arg Leu Gly Asp Thr Leu Tyr Glu Glu Phe Gly Ile           
            500                 505                 510                   

agc aaa taa gctt                                                      1549
Ser Lys                                                                   
                                                                          


<210>  10
<211>  514
<212>  PRT
<213>  Saccharomyces cerevisiae

<400>  10

Met Ala Ser Arg Gly Ser His His His His His His Gly Ala Met Thr 
1               5                   10                  15      


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


Ala Arg Lys Pro Ser Pro Leu Lys Thr Cys Ile His Leu Phe Gln Asp 
        35                  40                  45              


Pro Asn Ile Ile Phe Leu Gly Gly Gly Leu Pro Leu Lys Asp Tyr Phe 
    50                  55                  60                  


Pro Trp Asp Asn Leu Ser Val Asp Ser Pro Lys Pro Pro Phe Pro Gln 
65                  70                  75                  80  


Gly Ile Gly Ala Pro Ile Asp Glu Gln Asn Cys Ile Lys Tyr Thr Val 
                85                  90                  95      


Asn Lys Asp Tyr Ala Asp Lys Ser Ala Asn Pro Ser Asn Asp Ile Pro 
            100                 105                 110         


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


Leu Asn Phe Ile Arg Asp His Thr Lys Ile Ile His Asp Leu Lys Tyr 
    130                 135                 140                 


Lys Asp Trp Asp Val Leu Ala Thr Ala Gly Asn Thr Asn Ala Trp Glu 
145                 150                 155                 160 


Ser Thr Leu Arg Val Phe Cys Asn Arg Gly Asp Val Ile Leu Val Glu 
                165                 170                 175     


Ala His Ser Phe Ser Ser Ser Leu Ala Ser Ala Glu Ala Gln Gly Val 
            180                 185                 190         


Ile Thr Phe Pro Val Pro Ile Asp Ala Asp Gly Ile Ile Pro Glu Lys 
        195                 200                 205             


Leu Ala Lys Val Met Glu Asn Trp Thr Pro Gly Ala Pro Lys Pro Lys 
    210                 215                 220                 


Leu Leu Tyr Thr Ile Pro Thr Gly Gln Asn Pro Thr Gly Thr Ser Ile 
225                 230                 235                 240 


Ala Asp His Arg Lys Glu Ala Ile Tyr Lys Ile Ala Gln Lys Tyr Asp 
                245                 250                 255     


Phe Leu Ile Val Glu Asp Glu Pro Tyr Tyr Phe Leu Gln Met Asn Pro 
            260                 265                 270         


Tyr Ile Lys Asp Leu Lys Glu Arg Glu Lys Ala Gln Ser Ser Pro Lys 
        275                 280                 285             


Gln Asp His Asp Glu Phe Leu Lys Ser Leu Ala Asn Thr Phe Leu Ser 
    290                 295                 300                 


Leu Asp Thr Glu Gly Arg Val Ile Arg Met Asp Ser Phe Ser Lys Val 
305                 310                 315                 320 


Leu Ala Pro Gly Thr Arg Leu Gly Trp Ile Thr Gly Ser Ser Lys Ile 
                325                 330                 335     


Leu Lys Pro Tyr Leu Ser Leu His Glu Met Thr Ile Gln Ala Pro Ala 
            340                 345                 350         


Gly Phe Thr Gln Val Leu Val Asn Ala Thr Leu Ser Arg Trp Gly Gln 
        355                 360                 365             


Lys Gly Tyr Leu Asp Trp Leu Leu Gly Leu Arg His Glu Tyr Thr Leu 
    370                 375                 380                 


Lys Arg Asp Cys Ala Ile Asp Ala Leu Tyr Lys Tyr Leu Pro Gln Ser 
385                 390                 395                 400 


Asp Ala Phe Val Ile Asn Pro Pro Ile Ala Gly Met Phe Phe Thr Val 
                405                 410                 415     


Asn Ile Asp Ala Ser Val His Pro Glu Phe Lys Thr Lys Tyr Asn Ser 
            420                 425                 430         


Asp Pro Tyr Gln Leu Glu Gln Ser Leu Tyr His Lys Val Val Glu Arg 
        435                 440                 445             


Gly Val Leu Val Val Pro Gly Ser Trp Phe Lys Ser Glu Gly Glu Thr 
    450                 455                 460                 


Glu Pro Pro Gln Pro Ala Glu Ser Lys Glu Val Ser Asn Pro Asn Ile 
465                 470                 475                 480 


Ile Phe Phe Arg Gly Thr Tyr Ala Ala Val Ser Pro Glu Lys Leu Thr 
                485                 490                 495     


Glu Gly Leu Lys Arg Leu Gly Asp Thr Leu Tyr Glu Glu Phe Gly Ile 
            500                 505                 510         


Ser Lys 
        


<210>  11
<211>  1141
<212>  DNA
<213>  Bacillus sphaericus


<220>
<221>  CDS
<222>  (1)..(1137)
<223>  sequence of leudh of Bacillus sphaericus optimized for the codon 
       usage of Escherichia coli with an amino-terminal His6-tag

<220>
<221>  misc_feature
<222>  (37)..(42)
<223>  KasI restriction site

<220>
<221>  misc_feature
<222>  (1136)..(1141)
<223>  HindIII restriction site

<400>  11
atg gct agc aga gga tcg cat cac cat cac cat cac ggc gcc atg gaa         48
Met Ala Ser Arg Gly Ser His His His His His His Gly Ala Met Glu           
1               5                   10                  15                

att ttc aaa tat atg gaa aaa tat gat tat gaa cag ctg gtg ttc tgt         96
Ile Phe Lys Tyr Met Glu Lys Tyr Asp Tyr Glu Gln Leu Val Phe Cys           
            20                  25                  30                    

cag gat gaa gca agc ggt ctg aaa gca gtt att gca att cat gat acc        144
Gln Asp Glu Ala Ser Gly Leu Lys Ala Val Ile Ala Ile His Asp Thr           
        35                  40                  45                        

acc ctg ggt ccg gca ctg ggt ggt gca cgt atg tgg acc tat gca agc        192
Thr Leu Gly Pro Ala Leu Gly Gly Ala Arg Met Trp Thr Tyr Ala Ser           
    50                  55                  60                            

gaa gaa aat gca att gaa gat gca ctg cgt ctg gca cgt ggt atg acc        240
Glu Glu Asn Ala Ile Glu Asp Ala Leu Arg Leu Ala Arg Gly Met Thr           
65                  70                  75                  80            

tat aaa aac gca gca gca ggt ctg aat ctg ggt ggc ggt aaa acc gtt        288
Tyr Lys Asn Ala Ala Ala Gly Leu Asn Leu Gly Gly Gly Lys Thr Val           
                85                  90                  95                

att att ggt gat ccg ttc aaa gat aaa aac gaa gaa atg ttt cgt gcc        336
Ile Ile Gly Asp Pro Phe Lys Asp Lys Asn Glu Glu Met Phe Arg Ala           
            100                 105                 110                   

ctg ggt cgt ttt att cag ggt ctg aat ggt cgt tat att acc gca gaa        384
Leu Gly Arg Phe Ile Gln Gly Leu Asn Gly Arg Tyr Ile Thr Ala Glu           
        115                 120                 125                       

gat gtt ggc acc acc gtt ctg gat atg gat ctg att cat gaa gaa acc        432
Asp Val Gly Thr Thr Val Leu Asp Met Asp Leu Ile His Glu Glu Thr           
    130                 135                 140                           

acc tat gtg acc ggt att agt ccg gca ttt ggt agc agc ggt aat ccg        480
Thr Tyr Val Thr Gly Ile Ser Pro Ala Phe Gly Ser Ser Gly Asn Pro           
145                 150                 155                 160           

agt ccg gtt acc gcc tat ggt gtt tat cgt ggc atg aaa gca gca gca        528
Ser Pro Val Thr Ala Tyr Gly Val Tyr Arg Gly Met Lys Ala Ala Ala           
                165                 170                 175               

aaa gaa gcc ttt ggt agc gat agc ctg gaa ggc ctg aaa gtt agc gtg        576
Lys Glu Ala Phe Gly Ser Asp Ser Leu Glu Gly Leu Lys Val Ser Val           
            180                 185                 190                   

cag ggt ctg ggt aat gtt gca tat aaa ctg tgt gaa tat ctg cac aac        624
Gln Gly Leu Gly Asn Val Ala Tyr Lys Leu Cys Glu Tyr Leu His Asn           
        195                 200                 205                       

gaa ggt gca aaa ctg gtt gtt acc gat att aat cag gca gcc att gat        672
Glu Gly Ala Lys Leu Val Val Thr Asp Ile Asn Gln Ala Ala Ile Asp           
    210                 215                 220                           

cgt gtg gtg aat gat ttt gat gca att gcc gtt gca ccg gat gaa att        720
Arg Val Val Asn Asp Phe Asp Ala Ile Ala Val Ala Pro Asp Glu Ile           
225                 230                 235                 240           

tat gca caa gaa gtg gat atc ttt agc ccg tgt gcg ctg ggt gca att        768
Tyr Ala Gln Glu Val Asp Ile Phe Ser Pro Cys Ala Leu Gly Ala Ile           
                245                 250                 255               

ctg aat gat gaa acc att ccg cag ctg aaa gca aaa gtt att gcc ggt        816
Leu Asn Asp Glu Thr Ile Pro Gln Leu Lys Ala Lys Val Ile Ala Gly           
            260                 265                 270                   

agc gca aat aac caa ctg aaa gat agc cgt cat ggt gat tat ctg cat        864
Ser Ala Asn Asn Gln Leu Lys Asp Ser Arg His Gly Asp Tyr Leu His           
        275                 280                 285                       

gag ctg ggt att gtt tat gct ccg gat tat gtt att aat gcc ggt ggt        912
Glu Leu Gly Ile Val Tyr Ala Pro Asp Tyr Val Ile Asn Ala Gly Gly           
    290                 295                 300                           

gtg att aat gtt gcc gat gaa ctg tat ggt tat aat cgt gaa cgt gcc        960
Val Ile Asn Val Ala Asp Glu Leu Tyr Gly Tyr Asn Arg Glu Arg Ala           
305                 310                 315                 320           

atg aaa cgt gtg gat ggt att tat gat agc atc gag aaa atc ttt gcc       1008
Met Lys Arg Val Asp Gly Ile Tyr Asp Ser Ile Glu Lys Ile Phe Ala           
                325                 330                 335               

atc agc aaa cgt gat ggt att ccg acc tat gtt gca gca aat cgt ctg       1056
Ile Ser Lys Arg Asp Gly Ile Pro Thr Tyr Val Ala Ala Asn Arg Leu           
            340                 345                 350                   

gcc gaa gaa cgt att gca cgt gtt gca aaa agc cgt agc cag ttt ctg       1104
Ala Glu Glu Arg Ile Ala Arg Val Ala Lys Ser Arg Ser Gln Phe Leu           
        355                 360                 365                       

aaa aac gaa aaa aac att ctg aac ggt cgt taa gctt                      1141
Lys Asn Glu Lys Asn Ile Leu Asn Gly Arg                                   
    370                 375                                               


<210>  12
<211>  378
<212>  PRT
<213>  Bacillus sphaericus

<400>  12

Met Ala Ser Arg Gly Ser His His His His His His Gly Ala Met Glu 
1               5                   10                  15      


Ile Phe Lys Tyr Met Glu Lys Tyr Asp Tyr Glu Gln Leu Val Phe Cys 
            20                  25                  30          


Gln Asp Glu Ala Ser Gly Leu Lys Ala Val Ile Ala Ile His Asp Thr 
        35                  40                  45              


Thr Leu Gly Pro Ala Leu Gly Gly Ala Arg Met Trp Thr Tyr Ala Ser 
    50                  55                  60                  


Glu Glu Asn Ala Ile Glu Asp Ala Leu Arg Leu Ala Arg Gly Met Thr 
65                  70                  75                  80  


Tyr Lys Asn Ala Ala Ala Gly Leu Asn Leu Gly Gly Gly Lys Thr Val 
                85                  90                  95      


Ile Ile Gly Asp Pro Phe Lys Asp Lys Asn Glu Glu Met Phe Arg Ala 
            100                 105                 110         


Leu Gly Arg Phe Ile Gln Gly Leu Asn Gly Arg Tyr Ile Thr Ala Glu 
        115                 120                 125             


Asp Val Gly Thr Thr Val Leu Asp Met Asp Leu Ile His Glu Glu Thr 
    130                 135                 140                 


Thr Tyr Val Thr Gly Ile Ser Pro Ala Phe Gly Ser Ser Gly Asn Pro 
145                 150                 155                 160 


Ser Pro Val Thr Ala Tyr Gly Val Tyr Arg Gly Met Lys Ala Ala Ala 
                165                 170                 175     


Lys Glu Ala Phe Gly Ser Asp Ser Leu Glu Gly Leu Lys Val Ser Val 
            180                 185                 190         


Gln Gly Leu Gly Asn Val Ala Tyr Lys Leu Cys Glu Tyr Leu His Asn 
        195                 200                 205             


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


Arg Val Val Asn Asp Phe Asp Ala Ile Ala Val Ala Pro Asp Glu Ile 
225                 230                 235                 240 


Tyr Ala Gln Glu Val Asp Ile Phe Ser Pro Cys Ala Leu Gly Ala Ile 
                245                 250                 255     


Leu Asn Asp Glu Thr Ile Pro Gln Leu Lys Ala Lys Val Ile Ala Gly 
            260                 265                 270         


Ser Ala Asn Asn Gln Leu Lys Asp Ser Arg His Gly Asp Tyr Leu His 
        275                 280                 285             


Glu Leu Gly Ile Val Tyr Ala Pro Asp Tyr Val Ile Asn Ala Gly Gly 
    290                 295                 300                 


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


Met Lys Arg Val Asp Gly Ile Tyr Asp Ser Ile Glu Lys Ile Phe Ala 
                325                 330                 335     


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


Ala Glu Glu Arg Ile Ala Arg Val Ala Lys Ser Arg Ser Gln Phe Leu 
        355                 360                 365             


Lys Asn Glu Lys Asn Ile Leu Asn Gly Arg 
    370                 375             


<210>  13
<211>  1147
<212>  DNA
<213>  Thermoactinomyces intermedius


<220>
<221>  CDS
<222>  (1)..(1143)
<223>  sequence of phedh of Thermoactinomyces intermedius optimized for 
       the codon usage of Escherichia coli with an amino-terminal 
       His6-tag

<220>
<221>  misc_feature
<222>  (37)..(42)
<223>  KasI restriction site

<220>
<221>  misc_feature
<222>  (1142)..(1147)
<223>  HindIII restriction site

<400>  13
atg gct agc aga gga tcg cat cac cat cac cat cac ggc gcc atg cgt         48
Met Ala Ser Arg Gly Ser His His His His His His Gly Ala Met Arg           
1               5                   10                  15                

gat gtt ttt gaa atg atg gat cgt tat ggt cac gaa cag gtg att ttt         96
Asp Val Phe Glu Met Met Asp Arg Tyr Gly His Glu Gln Val Ile Phe           
            20                  25                  30                    

tgt cgt cat ccg cag aca ggt ctg aaa gca att att gca ctg cat aat        144
Cys Arg His Pro Gln Thr Gly Leu Lys Ala Ile Ile Ala Leu His Asn           
        35                  40                  45                        

acc acc gca ggt ccg gca ctg ggt ggt tgt cgt atg att ccg tat gca        192
Thr Thr Ala Gly Pro Ala Leu Gly Gly Cys Arg Met Ile Pro Tyr Ala           
    50                  55                  60                            

agc acc gat gaa gca ctg gaa gat gtt ctg cgt ctg agc aaa ggt atg        240
Ser Thr Asp Glu Ala Leu Glu Asp Val Leu Arg Leu Ser Lys Gly Met           
65                  70                  75                  80            

acc tat aaa tgt agc ctg gcc gat gtt gat ttt ggt ggt ggt aaa atg        288
Thr Tyr Lys Cys Ser Leu Ala Asp Val Asp Phe Gly Gly Gly Lys Met           
                85                  90                  95                

gtg att att ggc gat ccg aaa aaa gac aaa agt ccg gaa ctg ttt cgt        336
Val Ile Ile Gly Asp Pro Lys Lys Asp Lys Ser Pro Glu Leu Phe Arg           
            100                 105                 110                   

gtg att ggt cgt ttt gtt ggt ggt ctg aat ggt cgc ttt tat acc ggc        384
Val Ile Gly Arg Phe Val Gly Gly Leu Asn Gly Arg Phe Tyr Thr Gly           
        115                 120                 125                       

acc gat atg ggc acc aat ccg gaa gat ttt gtt cat gca gca cgt gaa        432
Thr Asp Met Gly Thr Asn Pro Glu Asp Phe Val His Ala Ala Arg Glu           
    130                 135                 140                           

agc aaa agt ttt gca ggt ctg ccg aaa agc tat ggt ggc aaa ggt gat        480
Ser Lys Ser Phe Ala Gly Leu Pro Lys Ser Tyr Gly Gly Lys Gly Asp           
145                 150                 155                 160           

acc agc att ccg acc gca ctg ggc gtt ttt cat ggt atg cgt gca acc        528
Thr Ser Ile Pro Thr Ala Leu Gly Val Phe His Gly Met Arg Ala Thr           
                165                 170                 175               

gca cgt ttt ctg tgg ggc acc gat cag ctg aaa ggt cgt gtt gtt gca        576
Ala Arg Phe Leu Trp Gly Thr Asp Gln Leu Lys Gly Arg Val Val Ala           
            180                 185                 190                   

att cag ggt gtt ggt aaa gtt ggt gaa cgt ctg ctg cag ctg ctg gtt        624
Ile Gln Gly Val Gly Lys Val Gly Glu Arg Leu Leu Gln Leu Leu Val           
        195                 200                 205                       

gaa gtt ggt gca tat tgt aaa att gcc gat att gat agc gtt cgt tgc        672
Glu Val Gly Ala Tyr Cys Lys Ile Ala Asp Ile Asp Ser Val Arg Cys           
    210                 215                 220                           

gag caa ctg aaa gaa aaa tat ggt gat aaa gtg cag ctg gtt gat gtg        720
Glu Gln Leu Lys Glu Lys Tyr Gly Asp Lys Val Gln Leu Val Asp Val           
225                 230                 235                 240           

aac cgt att cat aaa gaa agc tgc gat atc ttt agc ccg tgt gca aaa        768
Asn Arg Ile His Lys Glu Ser Cys Asp Ile Phe Ser Pro Cys Ala Lys           
                245                 250                 255               

ggt ggt gtg gtt aat gat gat acc att gat gaa ttt cgt tgc ctg gca        816
Gly Gly Val Val Asn Asp Asp Thr Ile Asp Glu Phe Arg Cys Leu Ala           
            260                 265                 270                   

att gtt ggt agc gca aat aat cag ctg gta gaa gat cgt cat ggt gca        864
Ile Val Gly Ser Ala Asn Asn Gln Leu Val Glu Asp Arg His Gly Ala           
        275                 280                 285                       

ctg ctg cag aaa cgt agc att tgt tat gca ccg gat tat ctg gtt aat        912
Leu Leu Gln Lys Arg Ser Ile Cys Tyr Ala Pro Asp Tyr Leu Val Asn           
    290                 295                 300                           

gca ggc ggt ctg att cag gtt gca gat gaa ctg gaa ggt ttt cat gaa        960
Ala Gly Gly Leu Ile Gln Val Ala Asp Glu Leu Glu Gly Phe His Glu           
305                 310                 315                 320           

gaa cgt gtt ctg gca aaa acc gaa gcc att tat gat atg gtg ctg gat       1008
Glu Arg Val Leu Ala Lys Thr Glu Ala Ile Tyr Asp Met Val Leu Asp           
                325                 330                 335               

att ttt cac cgt gcc aaa aac gaa aac att acc acc tgt gaa gca gcc       1056
Ile Phe His Arg Ala Lys Asn Glu Asn Ile Thr Thr Cys Glu Ala Ala           
            340                 345                 350                   

gat cgt att gtt atg gaa cgt ctg aaa aaa ctg acc gat att cgt cgt       1104
Asp Arg Ile Val Met Glu Arg Leu Lys Lys Leu Thr Asp Ile Arg Arg           
        355                 360                 365                       

att ctg ctg gaa gat ccg cgt aat agc gca cgt cgt taa gctt              1147
Ile Leu Leu Glu Asp Pro Arg Asn Ser Ala Arg Arg                           
    370                 375                 380                           


<210>  14
<211>  380
<212>  PRT
<213>  Thermoactinomyces intermedius

<400>  14

Met Ala Ser Arg Gly Ser His His His His His His Gly Ala Met Arg 
1               5                   10                  15      


Asp Val Phe Glu Met Met Asp Arg Tyr Gly His Glu Gln Val Ile Phe 
            20                  25                  30          


Cys Arg His Pro Gln Thr Gly Leu Lys Ala Ile Ile Ala Leu His Asn 
        35                  40                  45              


Thr Thr Ala Gly Pro Ala Leu Gly Gly Cys Arg Met Ile Pro Tyr Ala 
    50                  55                  60                  


Ser Thr Asp Glu Ala Leu Glu Asp Val Leu Arg Leu Ser Lys Gly Met 
65                  70                  75                  80  


Thr Tyr Lys Cys Ser Leu Ala Asp Val Asp Phe Gly Gly Gly Lys Met 
                85                  90                  95      


Val Ile Ile Gly Asp Pro Lys Lys Asp Lys Ser Pro Glu Leu Phe Arg 
            100                 105                 110         


Val Ile Gly Arg Phe Val Gly Gly Leu Asn Gly Arg Phe Tyr Thr Gly 
        115                 120                 125             


Thr Asp Met Gly Thr Asn Pro Glu Asp Phe Val His Ala Ala Arg Glu 
    130                 135                 140                 


Ser Lys Ser Phe Ala Gly Leu Pro Lys Ser Tyr Gly Gly Lys Gly Asp 
145                 150                 155                 160 


Thr Ser Ile Pro Thr Ala Leu Gly Val Phe His Gly Met Arg Ala Thr 
                165                 170                 175     


Ala Arg Phe Leu Trp Gly Thr Asp Gln Leu Lys Gly Arg Val Val Ala 
            180                 185                 190         


Ile Gln Gly Val Gly Lys Val Gly Glu Arg Leu Leu Gln Leu Leu Val 
        195                 200                 205             


Glu Val Gly Ala Tyr Cys Lys Ile Ala Asp Ile Asp Ser Val Arg Cys 
    210                 215                 220                 


Glu Gln Leu Lys Glu Lys Tyr Gly Asp Lys Val Gln Leu Val Asp Val 
225                 230                 235                 240 


Asn Arg Ile His Lys Glu Ser Cys Asp Ile Phe Ser Pro Cys Ala Lys 
                245                 250                 255     


Gly Gly Val Val Asn Asp Asp Thr Ile Asp Glu Phe Arg Cys Leu Ala 
            260                 265                 270         


Ile Val Gly Ser Ala Asn Asn Gln Leu Val Glu Asp Arg His Gly Ala 
        275                 280                 285             


Leu Leu Gln Lys Arg Ser Ile Cys Tyr Ala Pro Asp Tyr Leu Val Asn 
    290                 295                 300                 


Ala Gly Gly Leu Ile Gln Val Ala Asp Glu Leu Glu Gly Phe His Glu 
305                 310                 315                 320 


Glu Arg Val Leu Ala Lys Thr Glu Ala Ile Tyr Asp Met Val Leu Asp 
                325                 330                 335     


Ile Phe His Arg Ala Lys Asn Glu Asn Ile Thr Thr Cys Glu Ala Ala 
            340                 345                 350         


Asp Arg Ile Val Met Glu Arg Leu Lys Lys Leu Thr Asp Ile Arg Arg 
        355                 360                 365             


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


201600130
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