                         SEQUENCE LISTING

<110>  SANOFI S.A.
 
<120>  NOVEL CYTOCHROME P450 POLYPEPTIDE WITH INCREASED ENZYMATIC 
       ACTIVITY

<130>  BET 13P3205

<150>  EP 14305071.4
<151>  2014-01-20

<160>  8     

<170>  PatentIn version 3.5

<210>  1
<211>  481
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Mutated amino acid sequence of CYP11A1

<400>  1

Ile Ser Thr Lys Thr Pro Arg Pro Tyr Ser Glu Ile Pro Ser Pro Gly 
1               5                   10                  15      


Asp Asn Gly Trp Leu Asn Leu Tyr His Phe Trp Arg Glu Lys Gly Ser 
            20                  25                  30          


Gln Arg Ile His Phe Arg His Ile Glu Asn Phe Gln Lys Tyr Gly Pro 
        35                  40                  45              


Ile Tyr Arg Glu Lys Leu Gly Asn Leu Glu Ser Val Tyr Ile Ile His 
    50                  55                  60                  


Pro Glu Asp Val Ala His Leu Phe Lys Phe Glu Gly Ser Tyr Pro Glu 
65                  70                  75                  80  


Arg Tyr Asp Ile Pro Pro Trp Leu Ala Tyr His Arg Tyr Tyr Gln Lys 
                85                  90                  95      


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


Val Val Leu Asn Thr Glu Val Met Ala Pro Glu Ala Ile Lys Asn Phe 
        115                 120                 125             


Ile Pro Leu Leu Asn Pro Val Ser Gln Asp Phe Val Ser Leu Leu His 
    130                 135                 140                 


Lys Arg Ile Lys Gln Gln Gly Ser Gly Lys Phe Val Gly Asp Ile Lys 
145                 150                 155                 160 


Glu Asp Leu Phe His Phe Ala Phe Glu Ser Ile Thr Asn Val Met Phe 
                165                 170                 175     


Gly Glu Arg Leu Gly Met Leu Glu Glu Thr Val Asn Pro Glu Ala Gln 
            180                 185                 190         


Lys Phe Ile Asp Ala Val Tyr Lys Met Phe His Thr Ser Val Pro Leu 
        195                 200                 205             


Leu Asn Val Pro Pro Glu Leu Tyr Arg Leu Phe Arg Thr Lys Thr Trp 
    210                 215                 220                 


Thr Asp His Val Ala Ala Trp Asp Thr Ile Phe Asn Lys Ala Glu Lys 
225                 230                 235                 240 


Tyr Thr Glu Ile Phe Tyr Gln Asp Leu Arg Arg Lys Thr Glu Phe Arg 
                245                 250                 255     


Asn Tyr Pro Gly Ile Leu Tyr Cys Leu Leu Lys Ser Glu Lys Met Leu 
            260                 265                 270         


Leu Glu Asp Val Lys Ala Asn Ile Thr Glu Met Leu Ala Gly Gly Val 
        275                 280                 285             


Asp Thr Thr Ser Met Thr Leu Gln Trp His Leu Tyr Glu Met Ala Arg 
    290                 295                 300                 


Ser Leu Asn Val Gln Glu Met Leu Arg Glu Glu Val Leu Asn Ala Arg 
305                 310                 315                 320 


Arg Gln Ala Glu Gly Asp Ile Ser Lys Met Leu Gln Met Val Pro Leu 
                325                 330                 335     


Leu Lys Ala Ser Ile Lys Glu Thr Leu Arg Leu His Pro Ile Ser Val 
            340                 345                 350         


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


Ile Pro Ala Lys Thr Leu Val Gln Val Ala Ile Tyr Ala Met Gly Arg 
    370                 375                 380                 


Asp Pro Ala Phe Phe Ser Ser Pro Asp Lys Phe Asp Pro Thr Arg Trp 
385                 390                 395                 400 


Leu Ser Lys Asp Lys Asp Leu Ile His Phe Arg Asn Leu Gly Phe Gly 
                405                 410                 415     


Trp Gly Val Arg Gln Cys Val Gly Arg Arg Ile Ala Glu Leu Glu Met 
            420                 425                 430         


Thr Leu Phe Leu Ile His Ile Leu Glu Asn Phe Lys Val Glu Met Gln 
        435                 440                 445             


His Ile Gly Asp Val Asp Thr Ile Phe Asn Leu Ile Leu Thr Pro Asp 
    450                 455                 460                 


Lys Pro Ile Phe Leu Val Phe Arg Pro Phe Asn Gln Asp Pro Pro Gln 
465                 470                 475                 480 


Ala 
    


<210>  2
<211>  481
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Mutated amino acid sequence of CYP11A1

<400>  2

Ile Ser Thr Lys Thr Pro Arg Pro Tyr Ser Glu Ile Pro Ser Pro Gly 
1               5                   10                  15      


Asp Asn Gly Trp Leu Asn Leu Tyr His Phe Trp Arg Glu Lys Gly Ser 
            20                  25                  30          


Gln Arg Ile His Phe Arg His Ile Glu Asn Phe Gln Lys Tyr Gly Pro 
        35                  40                  45              


Ile Tyr Arg Glu Lys Leu Gly Asn Leu Glu Ser Val Tyr Ile Ile His 
    50                  55                  60                  


Pro Glu Asp Val Ala His Leu Phe Lys Phe Glu Gly Ser Tyr Pro Glu 
65                  70                  75                  80  


Arg Tyr Asp Ile Pro Pro Trp Leu Ala Tyr His Arg Tyr Tyr Gln Lys 
                85                  90                  95      


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


Val Val Leu Asn Thr Glu Val Met Ala Pro Glu Ala Ile Lys Asn Phe 
        115                 120                 125             


Ile Pro Leu Leu Asn Pro Val Ser Gln Asp Phe Val Ser Leu Leu His 
    130                 135                 140                 


Lys Arg Ile Lys Gln Gln Gly Ser Gly Lys Phe Val Gly Asp Ile Lys 
145                 150                 155                 160 


Glu Asp Leu Phe His Phe Ala Phe Glu Ser Ile Thr Asn Val Met Phe 
                165                 170                 175     


Gly Glu Arg Leu Gly Met Leu Glu Glu Thr Val Asn Pro Glu Ala Gln 
            180                 185                 190         


Lys Phe Ile Asp Ala Val Tyr Lys Met Phe His Thr Ser Val Pro Leu 
        195                 200                 205             


Leu Asn Val Pro Pro Glu Leu Tyr Arg Leu Phe Arg Thr Lys Thr Trp 
    210                 215                 220                 


Thr Asp His Val Ala Ala Trp Asp Thr Ile Phe Asn Lys Ala Glu Lys 
225                 230                 235                 240 


Tyr Thr Glu Ile Phe Tyr Gln Asp Leu Arg Arg Lys Thr Glu Phe Arg 
                245                 250                 255     


Asn Tyr Pro Gly Ile Leu Tyr Cys Leu Leu Lys Ser Glu Lys Met Leu 
            260                 265                 270         


Leu Glu Asp Val Lys Ala Asn Ile Thr Glu Met Leu Ala Gly Gly Val 
        275                 280                 285             


Asn Thr Thr Ser Met Thr Leu Gln Trp His Leu Tyr Glu Met Ala Arg 
    290                 295                 300                 


Ser Leu Asn Val Gln Glu Met Leu Arg Glu Glu Val Leu Asn Ala Arg 
305                 310                 315                 320 


Arg Gln Ala Glu Gly Asp Ile Ser Lys Met Leu Gln Met Val Pro Leu 
                325                 330                 335     


Leu Lys Ala Ser Ile Lys Glu Thr Leu Arg Leu His Pro Ile Ser Val 
            340                 345                 350         


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


Ile Pro Ala Lys Thr Leu Val Gln Val Ala Ile Tyr Ala Met Gly Arg 
    370                 375                 380                 


Asp Pro Ala Phe Phe Ser Ser Pro Asp Lys Phe Asp Pro Thr Arg Trp 
385                 390                 395                 400 


Leu Ser Lys Asp Lys Asp Leu Ile His Phe Arg Asn Leu Gly Phe Gly 
                405                 410                 415     


Trp Gly Val Arg Gln Cys Val Gly Arg Arg Ile Ala Glu Leu Glu Met 
            420                 425                 430         


Thr Leu Phe Leu Ile His Ile Leu Glu Asn Phe Lys Val Glu Met Gln 
        435                 440                 445             


His Ile Gly Asp Val Asp Thr Ile Phe Asn Leu Ile Leu Thr Pro Asp 
    450                 455                 460                 


Lys Pro Ile Phe Leu Val Phe Arg Pro Phe Asn Gln Asp Pro Pro Gln 
465                 470                 475                 480 


Ala 
    


<210>  3
<211>  481
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Mutated amino acid sequence of CYP11A1

<400>  3

Ile Ser Thr Lys Thr Pro Arg Pro Tyr Ser Glu Ile Pro Ser Pro Gly 
1               5                   10                  15      


Asp Asn Gly Trp Leu Asn Leu Tyr His Phe Trp Arg Glu Lys Gly Ser 
            20                  25                  30          


Gln Arg Ile His Phe Arg His Ile Glu Asn Phe Gln Lys Tyr Gly Pro 
        35                  40                  45              


Ile Tyr Arg Glu Lys Leu Gly Asn Leu Glu Ser Val Tyr Ile Ile His 
    50                  55                  60                  


Pro Glu Asp Val Ala His Leu Phe Lys Phe Glu Gly Ser Tyr Pro Glu 
65                  70                  75                  80  


Arg Tyr Asp Ile Pro Pro Trp Leu Ala Tyr His Arg Tyr Tyr Gln Lys 
                85                  90                  95      


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


Val Val Leu Asn Thr Glu Val Met Ala Pro Glu Ala Ile Lys Asn Phe 
        115                 120                 125             


Ile Pro Leu Leu Asn Pro Val Ser Gln Asp Phe Val Ser Leu Leu His 
    130                 135                 140                 


Lys Arg Ile Lys Gln Gln Gly Ser Gly Lys Phe Val Gly Asp Ile Lys 
145                 150                 155                 160 


Glu Asp Leu Phe His Phe Ala Phe Glu Ser Ile Thr Asn Val Met Phe 
                165                 170                 175     


Gly Glu Arg Leu Gly Met Leu Glu Glu Thr Val Asn Pro Glu Ala Gln 
            180                 185                 190         


Lys Phe Ile Asp Ala Val Tyr Lys Met Phe His Thr Ser Val Pro Leu 
        195                 200                 205             


Leu Asn Val Pro Pro Glu Leu Tyr Arg Leu Phe Arg Thr Lys Thr Trp 
    210                 215                 220                 


Arg Asp His Val Ala Ala Trp Asp Thr Ile Phe Asn Lys Ala Glu Lys 
225                 230                 235                 240 


Tyr Thr Glu Ile Phe Tyr Gln Asp Leu Arg Arg Lys Thr Glu Phe Arg 
                245                 250                 255     


Asn Tyr Pro Gly Ile Leu Tyr Cys Leu Leu Lys Ser Glu Lys Met Leu 
            260                 265                 270         


Leu Glu Asp Val Lys Ala Asn Ile Thr Glu Met Leu Ala Gly Gly Val 
        275                 280                 285             


Asp Thr Thr Ser Met Thr Leu Gln Trp His Leu Tyr Glu Met Ala Arg 
    290                 295                 300                 


Ser Leu Asn Val Gln Glu Met Leu Arg Glu Glu Val Leu Asn Ala Arg 
305                 310                 315                 320 


Arg Gln Ala Glu Gly Asp Ile Ser Lys Met Leu Gln Met Val Pro Leu 
                325                 330                 335     


Leu Lys Ala Ser Ile Lys Glu Thr Leu Arg Leu His Pro Ile Ser Val 
            340                 345                 350         


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


Ile Pro Ala Lys Thr Leu Val Gln Val Ala Ile Tyr Ala Met Gly Arg 
    370                 375                 380                 


Asp Pro Ala Phe Phe Ser Ser Pro Asp Lys Phe Asp Pro Thr Arg Trp 
385                 390                 395                 400 


Leu Ser Lys Asp Lys Asp Leu Ile His Phe Arg Asn Leu Gly Phe Gly 
                405                 410                 415     


Trp Gly Val Arg Gln Cys Val Gly Arg Arg Ile Ala Glu Leu Glu Met 
            420                 425                 430         


Thr Leu Phe Leu Ile His Ile Leu Glu Asn Phe Lys Val Glu Met Gln 
        435                 440                 445             


His Ile Gly Asp Val Asp Thr Ile Phe Asn Leu Ile Leu Thr Pro Asp 
    450                 455                 460                 


Lys Pro Ile Phe Leu Val Phe Arg Pro Phe Asn Gln Asp Pro Pro Gln 
465                 470                 475                 480 


Ala 
    


<210>  4
<211>  481
<212>  PRT
<213>  Bos taurus

<400>  4

Ile Ser Thr Lys Thr Pro Arg Pro Tyr Ser Glu Ile Pro Ser Pro Gly 
1               5                   10                  15      


Asp Asn Gly Trp Leu Asn Leu Tyr His Phe Trp Arg Glu Lys Gly Ser 
            20                  25                  30          


Gln Arg Ile His Phe Arg His Ile Glu Asn Phe Gln Lys Tyr Gly Pro 
        35                  40                  45              


Ile Tyr Arg Glu Lys Leu Gly Asn Leu Glu Ser Val Tyr Ile Ile His 
    50                  55                  60                  


Pro Glu Asp Val Ala His Leu Phe Lys Phe Glu Gly Ser Tyr Pro Glu 
65                  70                  75                  80  


Arg Tyr Asp Ile Pro Pro Trp Leu Ala Tyr His Arg Tyr Tyr Gln Lys 
                85                  90                  95      


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


Val Val Leu Asn Thr Glu Val Met Ala Pro Glu Ala Ile Lys Asn Phe 
        115                 120                 125             


Ile Pro Leu Leu Asn Pro Val Ser Gln Asp Phe Val Ser Leu Leu His 
    130                 135                 140                 


Lys Arg Ile Lys Gln Gln Gly Ser Gly Lys Phe Val Gly Asp Ile Lys 
145                 150                 155                 160 


Glu Asp Leu Phe His Phe Ala Phe Glu Ser Ile Thr Asn Val Met Phe 
                165                 170                 175     


Gly Glu Arg Leu Gly Met Leu Glu Glu Thr Val Asn Pro Glu Ala Gln 
            180                 185                 190         


Lys Phe Ile Asp Ala Val Tyr Lys Met Phe His Thr Ser Val Pro Leu 
        195                 200                 205             


Leu Asn Val Pro Pro Glu Leu Tyr Arg Leu Phe Arg Thr Lys Thr Trp 
    210                 215                 220                 


Arg Asp His Val Ala Ala Trp Asp Thr Ile Phe Asn Lys Ala Glu Lys 
225                 230                 235                 240 


Tyr Thr Glu Ile Phe Tyr Gln Asp Leu Arg Arg Lys Thr Glu Phe Arg 
                245                 250                 255     


Asn Tyr Pro Gly Ile Leu Tyr Cys Leu Leu Lys Ser Glu Lys Met Leu 
            260                 265                 270         


Leu Glu Asp Val Lys Ala Asn Ile Thr Glu Met Leu Ala Gly Gly Val 
        275                 280                 285             


Asn Thr Thr Ser Met Thr Leu Gln Trp His Leu Tyr Glu Met Ala Arg 
    290                 295                 300                 


Ser Leu Asn Val Gln Glu Met Leu Arg Glu Glu Val Leu Asn Ala Arg 
305                 310                 315                 320 


Arg Gln Ala Glu Gly Asp Ile Ser Lys Met Leu Gln Met Val Pro Leu 
                325                 330                 335     


Leu Lys Ala Ser Ile Lys Glu Thr Leu Arg Leu His Pro Ile Ser Val 
            340                 345                 350         


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


Ile Pro Ala Lys Thr Leu Val Gln Val Ala Ile Tyr Ala Met Gly Arg 
    370                 375                 380                 


Asp Pro Ala Phe Phe Ser Ser Pro Asp Lys Phe Asp Pro Thr Arg Trp 
385                 390                 395                 400 


Leu Ser Lys Asp Lys Asp Leu Ile His Phe Arg Asn Leu Gly Phe Gly 
                405                 410                 415     


Trp Gly Val Arg Gln Cys Val Gly Arg Arg Ile Ala Glu Leu Glu Met 
            420                 425                 430         


Thr Leu Phe Leu Ile His Ile Leu Glu Asn Phe Lys Val Glu Met Gln 
        435                 440                 445             


His Ile Gly Asp Val Asp Thr Ile Phe Asn Leu Ile Leu Thr Pro Asp 
    450                 455                 460                 


Lys Pro Ile Phe Leu Val Phe Arg Pro Phe Asn Gln Asp Pro Pro Gln 
465                 470                 475                 480 


Ala 
    


<210>  5
<211>  9745
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  plasmid

<400>  5
catggtctca cttttccact ttttgtcttg tccactaaaa cccttgattt ttcatctgaa       60

taaatgctac tattaggaca cataatatta aaagaaaccc ccatctattt agttatttgt      120

ttggtcactt ataactttaa cagatggggt ttttctgtgc aaccaatttt aagggttttc      180

aatactttaa aacacataca taccaacact tcaacgcacc tttcagcaac taaaataaaa      240

atgacgttat ttctatatgt atcaagataa gaaagaacaa gttcaaaacc atcaaaaaaa      300

gacacctttt caggtgcttt ttttatttta taaactcatt ccctgatctc gacttcgttc      360

tttttttacc tctcggttat gagttagttc aaattcgttc tttttaggtt ctaaatcgtg      420

tttttcttgg aattgtgctg ttttatcctt taccttgtct acaaacccct taaaaacgtt      480

tttaaaggct tttaagccgt ctgtacgttc cttaagatca acgtgatata ggtttgctaa      540

cctttgcgtt cacttaacta acttataggg gtaacactta aaaaagaatc aataacgata      600

gaaaccgctc ctaaagcagg tgcatttttt cctaacgaag aaggcaatag ttcacattta      660

ttgtctaaat gagaatggac tctagaagaa acttcgtttt taatcgtatt taaaacaatg      720

ggatgagatt caattatatg atttctcaag ataacagctt ctatatcaaa tgtattaagg      780

atattggtta atccaattcc gatataaaag ccaaagtttt gaagtgcatt taacatttct      840

acatcatttt tatttgcgcg ttccacaatc tcttttcgag aaatattctt ttcttcttta      900

gagagcgaag ccagtaacgc tttttcagaa gcatataatt cccaacagcc tcgatttcca      960

cagctgcatt tgggtccatt aaaatctatc gtcatatgac ccatttcccc agaaaaaccc     1020

tgaacacctt tatacaattc gttgttaata acaagtccag ttccaattcc gatattaata     1080

ctgatgtaaa cgatgttttc atagtttttt gtcataccaa atactttttc accgtatgct     1140

cctgcattag cttcattttc aacaaaaacc ggaacattaa actcactctc aattaaaaac     1200

tgcaaatctt tgatattcca atttaagtta ggcatgaaaa taatttgctg atgacgatct     1260

acaaggcctg gaacacaaat tcctattccg actagaccat aaggggactc aggcatatgg     1320

gttacaaaac catgaataag tgcaaataaa atctctttta cttcactagc ggaagaacta     1380

gacaagtcag aagtcttctc gagaataata tttccttcta agtcggttag aattccgtta     1440

agatagtcga ctcctatatc aataccaatc gagtagcctg cattcttatt aaaaacaagc     1500

attacaggtc ttctgccgcc tctagattgc cctgccccaa tttcaaaaat aaaatctttt     1560

tcaagcagtg tatttacttg agaggagaca gtagacttgt ttaatcctgt aatctcagag     1620

agagttgccc tggagacagg ggagttcttc aaaatttcat ctaatattaa tttttgattc     1680

atttttttta ctaaagcttg atctgcaatt tgaataataa ccactccttt gtttatccac     1740

cgaactaagt tggtgttttt tgaagcttga attagatatt taaaagtatc atatctaata     1800

ttataactaa attttctaaa aaaaacattg aaataaacat taaattaata tatgatggaa     1860

ttgtagttag tttacaattc caacaaacta actcaattaa gctagctgat ggataaactt     1920

gttcacttaa ttaaactagt aaatcaagga ggtgaatata caatgattag cacaaaaaca     1980

cctcgccctt attctgaaat tcctagccct ggtgataatg gatggttaaa tttatatcat     2040

ttttggcgtg aaaaaggtag ccaacgcatt cattttcgtc atattgaaaa ttttcaaaaa     2100

tatggaccta tttatcgcga aaaattagga aatttagaaa gcgtatatat tattcatcct     2160

gaagatgtag ctcatttatt taaatttgaa ggatcttatc ctgaacgcta tgatattcct     2220

ccttggcttg cttatcatcg ttattatcaa aaacctattg gcgtattatt taaaaaatct     2280

ggaacatgga aaaaagatcg tgtagtatta aatacagaag taatggctcc tgaagctatt     2340

aaaaatttta ttccgttatt aaatcctgta tctcaagatt ttgtatctct tttacataaa     2400

cgtattaaac aacaaggatc tggaaaattt gttggagaca ttaaagaaga tttatttcat     2460

tttgcgtttg aatctattac aaatgttatg tttggagaac gccttggaat gttagaagaa     2520

acggtaaacc ctgaagctca aaaatttatt gatgctgtat ataaaatgtt tcatacatct     2580

gtacctttat taaacgtacc tcctgaactt tatcgtcttt ttcgaacgaa aacgtggaca     2640

gatcatgtag ctgcttggga tacaattttt aataaagcgg aaaaatacac ggaaattttt     2700

tatcaagatt tacgtcgtaa aacagaattt cgtaattatc cgggaattct ttattgttta     2760

cttaaaagcg aaaagatgtt attagaagat gtaaaagcta atatcacaga aatgttagca     2820

ggtggagtag atacaacaag catgacatta caatggcatc tttatgaaat ggctcgcagc     2880

ttaaacgtac aagaaatgtt acgtgaagaa gtattaaacg ctcgtcgtca agctgaaggt     2940

gatatttcta aaatgttaca aatggttcca ttattaaaag cttctattaa agaaacgtta     3000

cgtttacatc caattagcgt aacgcttcaa cgttatcctg aatctgattt agtattacaa     3060

gattatctta ttcctgctaa aacattagta caagtagcta tttatgctat gggacgtgat     3120

cctgcttttt tttcttctcc tgataaattt gatcctacac gttggttatc taaagataaa     3180

gatcttattc attttcgcaa tcttggattt ggatggggag tacgtcaatg tgtaggacgt     3240

cgtattgctg aattagaaat gacgcttttt cttattcaca ttcttgaaaa ctttaaagtg     3300

gaaatgcaac atattggaga tgtagacacg atttttaact taattcttac gcctgataaa     3360

cctatttttt tagtatttcg tccttttaat caagatcctc ctcaagctta ataaacgcgt     3420

ggtaccaaat caaggaggtg aatatacaat gtctacacaa gaacaaacac ctcaaatttg     3480

tgtagtagga tctggacctg ctggatttta tacagctcaa catcttttaa aacatcattc     3540

tcgcgctcat gtagatattt atgaaaaaca acttgtacct tttggattag ttcgttttgg     3600

agtagctcct gatcatcctg aagtaaaaaa cgtaattaac acatttacac agacagctcg     3660

ttctgatcgt tgtgcttttt atggaaatgt agaagtagga cgtgatgtaa cagtacaaga     3720

acttcaagat gcttatcatg ctgtagtatt atcttatggt gctgaagatc atcaagcttt     3780

agatattcca ggtgaagaat tacctggtgt attttctgct cgtgcttttg taggatggta     3840

taatggatta cctgaaaatc gtgaattagc tcctgattta tcttgtgata cagctgtaat     3900

tttaggacaa ggcaacgtag ctttagatgt agctcgtatt ttattaacac ctccggatca     3960

tttagaaaaa acggatatta cagaagctgc tcttggagct ttacgtcaat ctcgtgtaaa     4020

aacagtatgg attgtaggac gtcgtggacc tttacaagta gcttttacga ttaaagaact     4080

tcgcgaaatg attcaattac ctggaacacg tcctatgtta gatcctgctg attttttagg     4140

acttcaggat cgtattaaag aagctgcacg tcctcgtaaa cgtttaatgg aattattatt     4200

acgtacagct acagaaaaac ctggtgtaga agaagctgct cgtagagcat ctgcttctcg     4260

tgcttgggga ttacgttttt ttcgtagccc tcaacaagta ttaccttctc ctgatggacg     4320

tcgtgctgct ggaattcgtt tagctgtaac acgtttagaa ggtattggag aagctacacg     4380

tgctgtacct acaggtgatg tagaagattt accttgtgga cttgtattaa gctctattgg     4440

atataaatct cgtcctattg atccttctgt accttttgat cctaaattag gtgtagtacc     4500

taatatggaa ggacgtgtag tagatgtacc tggattatat tgttctggat gggtaaaacg     4560

tggacctaca ggtgtaatta caacaacaat gacagatagc tttttaacag gccaaattct     4620

tttacaagat cttaaagctg gacatttacc ttcaggacct cgtcctggat ctgcttttat     4680

taaagcttta cttgattctc gtggagtatg gcctgtatct ttttctgatt gggaaaaatt     4740

agatgctgaa gaagtatcta gaggacaagc ttctggaaaa cctcgtgaaa aattattaga     4800

tcctcaagaa atgcttcgtt tacttggcca ctaataagag ctctgtacaa atcaaggagg     4860

tgaatataca atgtcttctt ctgaagataa aataacagtc cactttataa accgtgatgg     4920

tgaaacatta acaaccaaag gaaaaattgg tgactctctg ctagatgttg tggttcaaaa     4980

taatctagat attgatggtt ttggtgcatg tgagggaacc ttggcttgtt ctacctgtca     5040

cctcatcttt gaacagcaca tatttgagaa attggaagca atcactgatg aggagaatga     5100

catgcttgat ctggcatatg gactaacaga tagatcgcgg ttgggctgcc agatctgttt     5160

gacaaaggct atggacaata tgactgttcg agtaccatag catgcgcggc cgccatgccg     5220

gctaaacctc gcgaacggat tcaccggtcc aagaattgga gctaattaat tcttgcggag     5280

aactgtgaat gcgcaaacca acccttggca gaacatatcc atcgcgtccg ccatctccag     5340

cagccgcacg cggcgcatct cgggccgcgt tgctggcgtt tttccatagg ctccgccccc     5400

ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat     5460

aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc     5520

cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct     5580

cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg     5640

aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc     5700

cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga     5760

ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa     5820

ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta     5880

gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc     5940

agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg     6000

acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga     6060

tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg     6120

agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct     6180

gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg     6240

agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc     6300

cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa     6360

ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc     6420

cagttaatag tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt     6480

cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc     6540

ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt     6600

tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc     6660

catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt     6720

gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata     6780

gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga     6840

tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag     6900

catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa     6960

aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt     7020

attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga     7080

aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag     7140

aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc     7200

ttcaagaatt cctgttataa aaaaaggatc aattttgaac tctctcccaa agttgatccc     7260

ttaacgattt agaaatccct ttgagaatgt ttatatacat tcaaggtaac cagccaacta     7320

atgacaatga ttcctgaaaa aagtaataac aaattactat acagataagt tgactgatca     7380

acttccatag gtaacaacct ttgatcaagt aagggtatgg ataataaacc acctacaatt     7440

gcaatacctg ttccctctga taaaaagctg gtaaagttaa gcaaactcat tccagcacca     7500

gcttcctgct gtttcaagct acttgaaaca attgttgata taactgtttt ggtgaacgaa     7560

agcccaccta aaacaaatac gattataatt gtcatgaacc atgatgttgt ttctaaaaga     7620

aaggaagcag ttaaaaagct aacagaaaga aatgtaactc cgatgtttaa cacgtataaa     7680

ggacctcttc tatcaacaag tatcccacca atgtagccga aaataatgac actcattgtt     7740

ccagggaaaa taattacact tccgatttcg gcagtactta gctggtgaac atctttcatc     7800

atataaggaa ccatagagac aaaccctgct actgttccaa atataattcc cccacaaaga     7860

actccaatca taaaaggtat atttttccct aatccgggat caacaaaagg atctgttact     7920

ttcctgatat gttttacaaa tatcaggaat gacagcacgc taacgataag aaaagaaatg     7980

ctatatgatg ttgtaaacaa cataaaaaat acaatgccta cagacattag tataattcct     8040

ttgatatcaa aatgaccttt tatccttact tctttcttta ataatttcat aagaaacgga     8100

acagtgataa ttgttatcat aggaatgagt agaagatagg accaatgaat ataatgggct     8160

atcattccac caatcgctgg accgactcct tctcccatgg ctactatcga tccaataaga     8220

ccaaatgctt tacccctatt ttcctttgga atatagcgcg caactacaac cattacgagt     8280

gctggaaatg cagctgcacc agccccttga ataaaacgag ccataataag taaggaaaag     8340

aaagaatggc caacaaaccc aattaccgac ccgaaacaat ttattataat tccaaatagg     8400

agtaaccttt tgatgcctaa ttgatcagat agctttccat atacagctgt tccaatggaa     8460

aaggttaaca taaaggctgt gttcacccag tttgtactcg caggtggttt attaaaatca     8520

tttgcaatat caggtaatga gacgttcaaa accatttcat ttaatacgct aaaaaaagat     8580

aaaatgcaaa gccaaattaa aatttggttg tgtcgtaaat tcgattgtga ataggatgta     8640

ttcacatttc accctccaat aatgagggca gacgtagttt atagggttaa tgatacgctt     8700

ccctctttta attgaaccct gttacattca ttacacttca taattaattc ctcctaaact     8760

tgattaaaac attttaccac atataaacta agttttaaat tcagtatttc atcacttata     8820

caacaatatg gcccgtttgt tgaactactc tttaataaaa taatttttcc gttcccaatt     8880

ccacattgca ataatagaaa atccatcttc atcggctttt tcgtcatcat ctgtatgaat     8940

caaatcgcct tcttctgtgt catcaaggtt taatttttta tgtatttctt ttaacaaacc     9000

accataggag attaaccttt tacggtgtaa accttcctcc aaatcagaca aacgtttcaa     9060

attcttttct tcatcatcgg tcataaaatc cgtatccttt acaggatatt ttgcagtttc     9120

gtcaattgcc gattgtatat ccgatttata tttatttttc ggtcgaatca tttgaacttt     9180

tacatttgga tcatagtcta atttcattgc ctttttccaa aattgaatcc attgtttttg     9240

attcacgtag ttttctgtat tcttaaaata agttggttcc acacatacca atacatgcat     9300

gtgctgatta taagaattat ctttattatt tattgtcact tccgttgcac gcataaaacc     9360

aacaagattt ttattaattt ttttatattg catcattcgg cgaaatcctt gagccatatc     9420

tgacaaactc ttatttaatt cttcgccatc ataaacattt ttaactgtta atgtgagaaa     9480

caaccaacga actgttggct tttgtttaat aacttcagca acaacctttt gtgactgaat     9540

gccatgtttc attgctctcc tccagttgca cattggacaa agcctggatt tacaaaacca     9600

cactcgatac aactttcttt cgcctgtttc acgattttgt ttatactcta atatttcagc     9660

acaatctttt actctttcag cctttttaaa ttcaagaata tgcagaagtt caaagtaatc     9720

aacattagcg attttctttt ctctc                                           9745


<210>  6
<211>  481
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Mutated amino acid sequence of CYP11A1

<400>  6

Ala Ser Thr Lys Thr Pro Arg Pro Tyr Ser Glu Ile Pro Ser Pro Gly 
1               5                   10                  15      


Asp Asn Gly Trp Leu Asn Leu Tyr His Phe Trp Arg Glu Lys Gly Ser 
            20                  25                  30          


Gln Arg Ile His Phe Arg His Ile Glu Asn Phe Gln Lys Tyr Gly Pro 
        35                  40                  45              


Ile Tyr Arg Glu Lys Leu Gly Asn Leu Glu Ser Val Tyr Ile Ile His 
    50                  55                  60                  


Pro Glu Asp Val Ala His Leu Phe Lys Phe Glu Gly Ser Tyr Pro Glu 
65                  70                  75                  80  


Arg Tyr Asp Ile Pro Pro Trp Leu Ala Tyr His Arg Tyr Tyr Gln Lys 
                85                  90                  95      


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


Val Val Leu Asn Thr Glu Val Met Ala Pro Glu Ala Ile Lys Asn Phe 
        115                 120                 125             


Ile Pro Leu Leu Asn Pro Val Ser Gln Asp Phe Val Ser Leu Leu His 
    130                 135                 140                 


Lys Arg Ile Lys Gln Gln Gly Ser Gly Glu Phe Val Gly Asp Ile Lys 
145                 150                 155                 160 


Glu Asp Leu Phe His Phe Ala Phe Glu Ser Ile Thr Asn Val Met Phe 
                165                 170                 175     


Gly Glu Arg Leu Gly Met Leu Glu Glu Thr Val Asn Pro Glu Ala Gln 
            180                 185                 190         


Lys Phe Ile Asp Ala Val Tyr Lys Met Phe His Thr Ser Val Pro Leu 
        195                 200                 205             


Leu Asn Val Pro Pro Glu Leu Tyr Arg Leu Phe Arg Thr Lys Thr Trp 
    210                 215                 220                 


Arg Asp His Val Ala Ala Trp Asp Thr Ile Phe Asn Lys Ala Glu Lys 
225                 230                 235                 240 


Tyr Thr Glu Ile Phe Tyr Gln Asp Leu Arg Arg Lys Thr Glu Phe Arg 
                245                 250                 255     


Asn Tyr Pro Gly Ile Leu Tyr Cys Leu Leu Lys Ser Glu Lys Met Leu 
            260                 265                 270         


Leu Glu Asp Val Lys Ala Asn Ile Thr Glu Met Leu Ala Gly Gly Val 
        275                 280                 285             


Asn Thr Thr Ser Met Thr Leu Gln Trp His Leu Tyr Glu Met Ala Arg 
    290                 295                 300                 


Ser Leu Asn Val Gln Glu Met Leu Arg Glu Glu Val Leu Asn Ala Arg 
305                 310                 315                 320 


Arg Gln Ala Glu Gly Asp Ile Ser Lys Met Leu Gln Met Val Pro Leu 
                325                 330                 335     


Leu Lys Ala Ser Ile Lys Glu Thr Leu Arg Leu His Pro Ile Ser Val 
            340                 345                 350         


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


Ile Pro Ala Lys Thr Leu Val Gln Val Ala Ile Tyr Ala Met Gly Arg 
    370                 375                 380                 


Asp Pro Ala Phe Phe Ser Ser Pro Asp Lys Phe Asp Pro Thr Arg Trp 
385                 390                 395                 400 


Leu Ser Lys Asp Lys Asp Leu Ile His Phe Arg Asn Leu Gly Phe Gly 
                405                 410                 415     


Trp Gly Val Arg Gln Cys Val Gly Arg Arg Ile Ala Glu Leu Glu Met 
            420                 425                 430         


Thr Leu Phe Leu Ile His Ile Leu Glu Asn Phe Lys Val Glu Met Gln 
        435                 440                 445             


His Ile Gly Asp Val Asp Thr Ile Phe Asn Leu Ile Leu Thr Pro Asp 
    450                 455                 460                 


Lys Pro Ile Phe Leu Val Phe Arg Pro Phe Asn Gln Asp Pro Pro Gln 
465                 470                 475                 480 


Ala 
    


<210>  7
<211>  461
<212>  PRT
<213>  Bos taurus

<400>  7

Met Ser Thr Gln Glu Gln Thr Pro Gln Ile Cys Val Val Gly Ser Gly 
1               5                   10                  15      


Pro Ala Gly Phe Tyr Thr Ala Gln His Leu Leu Lys His His Ser Arg 
            20                  25                  30          


Ala His Val Asp Ile Tyr Glu Lys Gln Leu Val Pro Phe Gly Leu Val 
        35                  40                  45              


Arg Phe Gly Val Ala Pro Asp His Pro Glu Val Lys Asn Val Ile Asn 
    50                  55                  60                  


Thr Phe Thr Gln Thr Ala Arg Ser Asp Arg Cys Ala Phe Tyr Gly Asn 
65                  70                  75                  80  


Val Glu Val Gly Arg Asp Val Thr Val Gln Glu Leu Gln Asp Ala Tyr 
                85                  90                  95      


His Ala Val Val Leu Ser Tyr Gly Ala Glu Asp His Gln Ala Leu Asp 
            100                 105                 110         


Ile Pro Gly Glu Glu Leu Pro Gly Val Phe Ser Ala Arg Ala Phe Val 
        115                 120                 125             


Gly Trp Tyr Asn Gly Leu Pro Glu Asn Arg Glu Leu Ala Pro Asp Leu 
    130                 135                 140                 


Ser Cys Asp Thr Ala Val Ile Leu Gly Gln Gly Asn Val Ala Leu Asp 
145                 150                 155                 160 


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


Ile Thr Glu Ala Ala Leu Gly Ala Leu Arg Gln Ser Arg Val Lys Thr 
            180                 185                 190         


Val Trp Ile Val Gly Arg Arg Gly Pro Leu Gln Val Ala Phe Thr Ile 
        195                 200                 205             


Lys Glu Leu Arg Glu Met Ile Gln Leu Pro Gly Thr Arg Pro Met Leu 
    210                 215                 220                 


Asp Pro Ala Asp Phe Leu Gly Leu Gln Asp Arg Ile Lys Glu Ala Ala 
225                 230                 235                 240 


Arg Pro Arg Lys Arg Leu Met Glu Leu Leu Leu Arg Thr Ala Thr Glu 
                245                 250                 255     


Lys Pro Gly Val Glu Glu Ala Ala Arg Arg Ala Ser Ala Ser Arg Ala 
            260                 265                 270         


Trp Gly Leu Arg Phe Phe Arg Ser Pro Gln Gln Val Leu Pro Ser Pro 
        275                 280                 285             


Asp Gly Arg Arg Ala Ala Gly Ile Arg Leu Ala Val Thr Arg Leu Glu 
    290                 295                 300                 


Gly Ile Gly Glu Ala Thr Arg Ala Val Pro Thr Gly Asp Val Glu Asp 
305                 310                 315                 320 


Leu Pro Cys Gly Leu Val Leu Ser Ser Ile Gly Tyr Lys Ser Arg Pro 
                325                 330                 335     


Ile Asp Pro Ser Val Pro Phe Asp Pro Lys Leu Gly Val Val Pro Asn 
            340                 345                 350         


Met Glu Gly Arg Val Val Asp Val Pro Gly Leu Tyr Cys Ser Gly Trp 
        355                 360                 365             


Val Lys Arg Gly Pro Thr Gly Val Ile Thr Thr Thr Met Thr Asp Ser 
    370                 375                 380                 


Phe Leu Thr Gly Gln Ile Leu Leu Gln Asp Leu Lys Ala Gly His Leu 
385                 390                 395                 400 


Pro Ser Gly Pro Arg Pro Gly Ser Ala Phe Ile Lys Ala Leu Leu Asp 
                405                 410                 415     


Ser Arg Gly Val Trp Pro Val Ser Phe Ser Asp Trp Glu Lys Leu Asp 
            420                 425                 430         


Ala Glu Glu Val Ser Arg Gly Gln Ala Ser Gly Lys Pro Arg Glu Lys 
        435                 440                 445             


Leu Leu Asp Pro Gln Glu Met Leu Arg Leu Leu Gly His 
    450                 455                 460     


<210>  8
<211>  109
<212>  PRT
<213>  Bos taurus

<400>  8

Met Ser Ser Ser Glu Asp Lys Ile Thr Val His Phe Ile Asn Arg Asp 
1               5                   10                  15      


Gly Glu Thr Leu Thr Thr Lys Gly Lys Ile Gly Asp Ser Leu Leu Asp 
            20                  25                  30          


Val Val Val Gln Asn Asn Leu Asp Ile Asp Gly Phe Gly Ala Cys Glu 
        35                  40                  45              


Gly Thr Leu Ala Cys Ser Thr Cys His Leu Ile Phe Glu Gln His Ile 
    50                  55                  60                  


Phe Glu Lys Leu Glu Ala Ile Thr Asp Glu Glu Asn Asp Met Leu Asp 
65                  70                  75                  80  


Leu Ala Tyr Gly Leu Thr Asp Arg Ser Arg Leu Gly Cys Gln Ile Cys 
                85                  90                  95      


Leu Thr Lys Ala Met Asp Asn Met Thr Val Arg Val Pro 
            100                 105                 


