                         SEQUENCE LISTING

<110>  E. I. du Pont de Nemours and Company
       
       
       
 
<120>  A Butanol Dehydrogenase enzyme from the bacterium Achromobacter 
       xylosoxidans

<130>  CL3926

<160>  35    

<170>  PatentIn version 3.4

<210>  1
<211>  1047
<212>  DNA
<213>  Achromobacter xylosoxidans

<400>  1
atgaaagctc tggtttatca cggtgaccac aagatctcgc ttgaagacaa gcccaagccc     60

acccttcaaa agcccacgga tgtagtagta cgggttttga agaccacgat ctgcggcacg    120

gatctcggca tctacaaagg caagaatcca gaggtcgccg acgggcgcat cctgggccat    180

gaaggggtag gcgtcatcga ggaagtgggc gagagtgtca cgcagttcaa gaaaggcgac    240

aaggtcctga tttcctgcgt cacttcttgc ggctcgtgcg actactgcaa gaagcagctt    300

tactcccatt gccgcgacgg cgggtggatc ctgggttaca tgatcgatgg cgtgcaggcc    360

gaatacgtcc gcatcccgca tgccgacaac agcctctaca agatccccca gacaattgac    420

gacgaaatcg ccgtcctgct gagcgacatc ctgcccaccg gccacgaaat cggcgtccag    480

tatgggaatg tccagccggg cgatgcggtg gctattgtcg gcgcgggccc cgtcggcatg    540

tccgtactgt tgaccgccca gttctactcc ccctcgacca tcatcgtgat cgacatggac    600

gagaatcgcc tccagctcgc caaggagctc ggggcaacgc acaccatcaa ctccggcacg    660

gagaacgttg tcgaagccgt gcataggatt gcggcagagg gagtcgatgt tgcgatcgag    720

gcggtgggca taccggcgac ttgggacatc tgccaggaga tcgtcaagcc cggcgcgcac    780

atcgccaacg tcggcgtgca tggcgtcaag gttgacttcg agattcagaa gctctggatc    840

aagaacctga cgatcaccac gggactggtg aacacgaaca cgacgcccat gctgatgaag    900

gtcgcctcga ccgacaagct tccgttgaag aagatgatta cccatcgctt cgagctggcc    960

gagatcgagc acgcctatca ggtattcctc aatggcgcca aggagaaggc gatgaagatc   1020

atcctctcga acgcaggcgc tgcctga                                       1047


<210>  2
<211>  348
<212>  PRT
<213>  Achromobacter xylosoxidans

<400>  2

Met Lys Ala Leu Val Tyr His Gly Asp His Lys Ile Ser Leu Glu Asp 
1               5                   10                  15      


Lys Pro Lys Pro Thr Leu Gln Lys Pro Thr Asp Val Val Val Arg Val 
            20                  25                  30          


Leu Lys Thr Thr Ile Cys Gly Thr Asp Leu Gly Ile Tyr Lys Gly Lys 
        35                  40                  45              


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


Val Ile Glu Glu Val Gly Glu Ser Val Thr Gln Phe Lys Lys Gly Asp 
65                  70                  75                  80  


Lys Val Leu Ile Ser Cys Val Thr Ser Cys Gly Ser Cys Asp Tyr Cys 
                85                  90                  95      


Lys Lys Gln Leu Tyr Ser His Cys Arg Asp Gly Gly Trp Ile Leu Gly 
            100                 105                 110         


Tyr Met Ile Asp Gly Val Gln Ala Glu Tyr Val Arg Ile Pro His Ala 
        115                 120                 125             


Asp Asn Ser Leu Tyr Lys Ile Pro Gln Thr Ile Asp Asp Glu Ile Ala 
    130                 135                 140                 


Val Leu Leu Ser Asp Ile Leu Pro Thr Gly His Glu Ile Gly Val Gln 
145                 150                 155                 160 


Tyr Gly Asn Val Gln Pro Gly Asp Ala Val Ala Ile Val Gly Ala Gly 
                165                 170                 175     


Pro Val Gly Met Ser Val Leu Leu Thr Ala Gln Phe Tyr Ser Pro Ser 
            180                 185                 190         


Thr Ile Ile Val Ile Asp Met Asp Glu Asn Arg Leu Gln Leu Ala Lys 
        195                 200                 205             


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


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


Ala Val Gly Ile Pro Ala Thr Trp Asp Ile Cys Gln Glu Ile Val Lys 
                245                 250                 255     


Pro Gly Ala His Ile Ala Asn Val Gly Val His Gly Val Lys Val Asp 
            260                 265                 270         


Phe Glu Ile Gln Lys Leu Trp Ile Lys Asn Leu Thr Ile Thr Thr Gly 
        275                 280                 285             


Leu Val Asn Thr Asn Thr Thr Pro Met Leu Met Lys Val Ala Ser Thr 
    290                 295                 300                 


Asp Lys Leu Pro Leu Lys Lys Met Ile Thr His Arg Phe Glu Leu Ala 
305                 310                 315                 320 


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


Ala Met Lys Ile Ile Leu Ser Asn Ala Gly Ala Ala 
            340                 345             


<210>  3
<211>  1047
<212>  DNA
<213>  artificial sequence

<220>
<223>  codon optimized coding region

<400>  3
atgaaggcat tagtttatca tggggatcac aaaatttcgt tagaagacaa accaaaaccc     60

actctgcaga aaccaacaga cgttgtggtt agggtgttga aaacaacaat ttgcggtact    120

gacttgggaa tatacaaagg taagaatcct gaagtggcag atggcagaat cctgggtcat    180

gagggcgttg gcgtcattga agaagtgggc gaatccgtga cacaattcaa aaagggggat    240

aaagttttaa tctcctgcgt tactagctgt ggatcgtgtg attattgcaa gaagcaactg    300

tattcacact gtagagacgg tggctggatt ttaggttaca tgatcgacgg tgtccaagcc    360

gaatacgtca gaataccaca tgctgacaat tcattgtata agatcccgca aactatcgat    420

gatgaaattg cagtactact gtccgatatt ttacctactg gacatgaaat tggtgttcaa    480

tatggtaacg ttcaaccagg cgatgctgta gcaattgtag gagcaggtcc tgttggaatg    540

tcagttttgt taactgctca attttactcg cctagtacca ttattgttat cgacatggac    600

gaaaaccgtt tacaattagc gaaggagctt ggggccacac acactattaa ctccggtact    660

gaaaatgttg tcgaagctgt gcatcgtata gcagccgaag gagtggatgt agcaatagaa    720

gctgttggta tacccgcaac ctgggacatc tgtcaggaaa ttgtaaaacc cggcgctcat    780

attgccaacg tgggagttca tggtgttaag gtggactttg aaattcaaaa gttgtggatt    840

aagaatctaa ccatcaccac tggtttggtt aacactaata ctaccccaat gttgatgaag    900

gtagcctcta ctgataaatt gcctttaaag aaaatgatta ctcacaggtt tgagttagct    960

gaaatcgaac acgcatatca ggttttcttg aatggcgcta aagaaaaagc tatgaagatt   1020

attctatcta atgcaggtgc cgcctaa                                       1047


<210>  4
<211>  15
<212>  DNA
<213>  artificial sequence

<220>
<223>  primer

<400>  4
acgggcggtg tgtac                                                      15


<210>  5
<211>  16
<212>  DNA
<213>  artificial sequence

<220>
<223>  primer

<400>  5
gccagcagcc gcggta                                                     16


<210>  6
<211>  1756
<212>  DNA
<213>  Achromobacter xylosoxidans

<400>  6
taggacgtgc aattgagatc agctcgaatc gatcgctctg atcgttccct acggtgtggc     60

acttctcaga tcaatgataa cgtctaaccc ctaccgaacg gccctgaggt ttgacgctat    120

cctcttcttc ggtcgtcacg tcgaatttca ctagcagtcc tctttgtaag caattaattg    180

acaaggtttg cgttgtggag ggaacttcac caactctcca cgaccgaagc ggacggacag    240

ccaggaggct ctgggtttct gtttctctga ggagcaaagc aaaatttttt ggccccctta    300

ctcttttgtt tacgctcacc cagtcatgaa tcctaccgtg gtaatcgccc cccttgcggt    360

taggctaact acttctggta aaacccactc ccatggtgtg acgggcggtg tgtacaagac    420

ccgggaacgt attcaccgcg acatgctgat ccgcgattac tagcgattcc gacttcacgc    480

agtcgagttg cagactgcga tccggactac gatcgggttt ctgggattgg ctccccctcg    540

cgggttggcg accctctgtc ccgaccattg tatgacgtgt gaagccctac ccataagggc    600

catgaggact tgacgtcatc cccaccttcc tccggtttgt caccggcagt ctcattagag    660

tgccctttcg tagcaactaa tgacaagggt tgcgctcgtt gcgggactta acccaacatc    720

tcacgacacg agctgacgac agccatgcag cacctgtgtt ccggttctct tgcgagcact    780

gccaaatctc ttcggcattc cagacatgtc aagggtaggt aaggtttttc gcgttgcatc    840

gaattaatcc acatcatcca ccgcttgtgc gggtccccgt caattccttt gagttttaat    900

cttgcgaccg tactccccag gcggtcaact tcacgcgtta gctgcgctac caaggtccga    960

agaccccaac agctagttga catcgtttag ggcgtggact accagggtat ctaatcctgt   1020

ttgctcccca cgctttcgtg catgagcgtc agtgttatcc caggaggctg ccttcgccat   1080

cggtgttcct ccgcatatct acgcatttca ctgctacacg cggaattcca cctccctctg   1140

acacactcta gcccggtagt taaaaatgca gttccaaagt taagctctgg gatttcacat   1200

ctttctttcc gaaccgcctg cgcacgcttt acgcccagta attccgatta acgcttgcac   1260

cctacgtatt accgcggctg ctggcacgta gttagccggt gcttattctg caggtaccgt   1320

cagtttcgcg gggtattaac ccacgacgtt tctttcctgc caaaagtgct ttacaacccg   1380

aaggccttca tcgcacacgc gggatggctg gatcagggtt tcccccattg tccaaaattc   1440

cccactgctg cctcccgtag gagtctgggc cgtgtctcag tcccagtgtg gctggtcgtc   1500

ctctcaaacc agctacggat cgtcgccttg gtgagccgtt accccaccaa ctagctaatc   1560

cgatatcggc cgctccaata gtgcaaggtc ttgcgatccc ctgctttccc ccgtagggcg   1620

tatgcggtat tagctacgct ttcgcgtagt tatcccccgc tactgggcac gttccgatac   1680

attactcacc cgttcgccac tcgccaccag accgaagtcc gtgctgccgt tcgactgcat   1740

gttaagctcc gctgcc                                                   1756


<210>  7
<211>  20
<212>  PRT
<213>  Achromobacter xylosoxidans

<400>  7

Met Lys Ala Leu Val Tyr His Gly Asp His Lys Ile Ser Leu Gly Asp 
1               5                   10                  15      


Lys Pro Lys Pro 
            20  


<210>  8
<211>  21
<212>  DNA
<213>  artificial sequence

<220>
<223>  degenerate primer


<220>
<221>  misc_feature
<222>  (12)..(12)
<223>  n = a, t, g, or c

<220>
<221>  misc_feature
<222>  (15)..(15)
<223>  n = a, t, g, or c

<220>
<221>  misc_feature
<222>  (18)..(18)
<223>  n = c or t

<220>
<221>  misc_feature
<222>  (21)..(21)
<223>  n = c or t

<400>  8
atgaaagctc tngtntanca n                                               21


<210>  9
<211>  29
<212>  DNA
<213>  artificial sequence

<220>
<223>  primer

<400>  9
ggatctcggc atctacaaag gcacgaatc                                       29


<210>  10
<211>  29
<212>  DNA
<213>  artificial sequence

<220>
<223>  primer

<400>  10
ggattcgtgc ctttgtagat gccgagatc                                       29


<210>  11
<211>  27
<212>  DNA
<213>  artificial sequence

<220>
<223>  primer

<400>  11
ccagtgttag gattgatgct gtctccg                                         27


<210>  12
<211>  24
<212>  DNA
<213>  artificial sequence

<220>
<223>  primer

<400>  12
ggtgtttcgt gctgcgcatt aacg                                            24


<210>  13
<211>  24
<212>  DNA
<213>  artificial sequence

<220>
<223>  primer

<400>  13
ggtgtttcgt gctgcgcatt aacg                                            24


<210>  14
<211>  30
<212>  DNA
<213>  artificial sequence

<220>
<223>  primer

<400>  14
catgaagggg taggcgtcat cgaggaagtg                                      30


<210>  15
<211>  20
<212>  DNA
<213>  artificial sequence

<220>
<223>  primer

<400>  15
gcttgaagac aagcccaagc                                                 20


<210>  16
<211>  20
<212>  DNA
<213>  artificial sequence

<220>
<223>  primer

<400>  16
ggcaatggga gtaaagctgc                                                 20


<210>  17
<211>  37
<212>  DNA
<213>  artificial sequence

<220>
<223>  primer

<400>  17
cctgatttcc tgcgtcactt cttgcggctc gtgcgac                              37


<210>  18
<211>  23
<212>  DNA
<213>  artificial sequence

<220>
<223>  primer

<400>  18
cgaacacgac gcccatgctg atg                                             23


<210>  19
<211>  22
<212>  DNA
<213>  artificial sequence

<220>
<223>  primer

<400>  19
ggacatctgc caggagatcg tc                                              22


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

<220>
<223>  primer

<400>  20
caactccggc acggagaacg ttg                                             23


<210>  21
<211>  23
<212>  DNA
<213>  artificial sequence

<220>
<223>  primer

<400>  21
gcttctgaat ctcgaagtca acc                                             23


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

<220>
<223>  primer

<400>  22
cgagctggag gcgattctcg tcc                                             23


<210>  23
<211>  28
<212>  DNA
<213>  artificial sequence

<220>
<223>  primer

<400>  23
gcgtccaggg cgtcaaagat caggcagc                                        28


<210>  24
<211>  30
<212>  DNA
<213>  artificial sequence

<220>
<223>  primer

<400>  24
ggaattcaca catgaaagct ctggtttatc                                      30


<210>  25
<211>  16
<212>  DNA
<213>  artificial sequence

<220>
<223>  primer

<400>  25
gtaaaacgac ggccag                                                     16


<210>  26
<211>  22
<212>  DNA
<213>  artificial sequence

<220>
<223>  primer

<400>  26
cacacaggaa acagctatga cc                                              22


<210>  27
<211>  55
<212>  DNA
<213>  artificial sequence

<220>
<223>  primer

<400>  27
aaacaaacac acatattaca atagctgagg atgaaagctc tggtttatca cggtg          55


<210>  28
<211>  55
<212>  DNA
<213>  artificial sequence

<220>
<223>  primer

<400>  28
atcataagaa attcgcttac tcttaattaa tcaggcagcg cctgcgttcg agagg          55


<210>  29
<211>  753
<212>  DNA
<213>  Saccharomyces cerevisiae

<400>  29
gcatgcttgc atttagtcgt gcaatgtatg actttaagat ttgtgagcag gaagaaaagg     60

gagaatcttc taacgataaa cccttgaaaa actgggtaga ctacgctatg ttgagttgct    120

acgcaggctg cacaattaca cgagaatgct cccgcctagg atttaaggct aagggacgtg    180

caatgcagac gacagatcta aatgaccgtg tcggtgaagt gttcgccaaa cttttcggtt    240

aacacatgca gtgatgcacg cgcgatggtg ctaagttaca tatatatata tatagccata    300

gtgatgtcta agtaaccttt atggtatatt tcttaatgtg gaaagatact agcgcgcgca    360

cccacacaca agcttcgtct tttcttgaag aaaagaggaa gctcgctaaa tgggattcca    420

ctttccgttc cctgccagct gatggaaaaa ggttagtgga acgatgaaga ataaaaagag    480

agatccactg aggtgaaatt tcagctgaca gcgagtttca tgatcgtgat gaacaatggt    540

aacgagttgt ggctgttgcc agggagggtg gttctcaact tttaatgtat ggccaaatcg    600

ctacttgggt ttgttatata acaaagaaga aataatgaac tgattctctt cctccttctt    660

gtcctttctt aattctgttg taattacctt cctttgtaat tttttttgta attattcttc    720

ttaataatcc aaacaaacac acatattaca ata                                 753


<210>  30
<211>  316
<212>  DNA
<213>  Saccharomyces cerevisiae

<400>  30
gagtaagcga atttcttatg atttatgatt tttattatta aataagttat aaaaaaaata     60

agtgtataca aattttaaag tgactcttag gttttaaaac gaaaattctt attcttgagt    120

aactctttcc tgtaggtcag gttgctttct caggtatagc atgaggtcgc tcttattgac    180

cacacctcta ccggcatgcc gagcaaatgc ctgcaaatcg ctccccattt cacccaattg    240

tagatatgct aactccagca atgagttgat gaatctcggt gtgtatttta tgtcctcaga    300

ggacaacacc tgtggt                                                    316


<210>  31
<211>  1662
<212>  DNA
<213>  Lactococcus lactis

<400>  31
tctagacata tgtatactgt gggggattac ctgctggatc gcctgcacga actggggatt     60

gaagaaattt tcggtgtgcc aggcgattat aacctgcagt tcctggacca gattatctcg    120

cacaaagata tgaagtgggt cggtaacgcc aacgaactga acgcgagcta tatggcagat    180

ggttatgccc gtaccaaaaa agctgctgcg tttctgacga cctttggcgt tggcgaactg    240

agcgccgtca acggactggc aggaagctac gccgagaacc tgccagttgt cgaaattgtt    300

gggtcgccta cttctaaggt tcagaatgaa ggcaaatttg tgcaccatac tctggctgat    360

ggggatttta aacattttat gaaaatgcat gaaccggtta ctgcggcccg cacgctgctg    420

acagcagaga atgctacggt tgagatcgac cgcgtcctgt ctgcgctgct gaaagagcgc    480

aagccggtat atatcaatct gcctgtcgat gttgccgcag cgaaagccga aaagccgtcg    540

ctgccactga aaaaagaaaa cagcacctcc aatacatcgg accaggaaat tctgaataaa    600

atccaggaat cactgaagaa tgcgaagaaa ccgatcgtca tcaccggaca tgagatcatc    660

tcttttggcc tggaaaaaac ggtcacgcag ttcatttcta agaccaaact gcctatcacc    720

accctgaact tcggcaaatc tagcgtcgat gaagcgctgc cgagttttct gggtatctat    780

aatggtaccc tgtccgaacc gaacctgaaa gaattcgtcg aaagcgcgga ctttatcctg    840

atgctgggcg tgaaactgac ggatagctcc acaggcgcat ttacccacca tctgaacgag    900

aataaaatga tttccctgaa tatcgacgaa ggcaaaatct ttaacgagcg catccagaac    960

ttcgattttg aatctctgat tagttcgctg ctggatctgt ccgaaattga gtataaaggt   1020

aaatatattg ataaaaaaca ggaggatttt gtgccgtcta atgcgctgct gagtcaggat   1080

cgtctgtggc aagccgtaga aaacctgaca cagtctaatg aaacgattgt tgcggaacag   1140

ggaacttcat ttttcggcgc ctcatccatt tttctgaaat ccaaaagcca tttcattggc   1200

caaccgctgt gggggagtat tggttatacc tttccggcgg cgctgggttc acagattgca   1260

gataaggaat cacgccatct gctgtttatt ggtgacggca gcctgcagct gactgtccag   1320

gaactggggc tggcgatccg tgaaaaaatc aatccgattt gctttatcat caataacgac   1380

ggctacaccg tcgaacgcga aattcatgga ccgaatcaaa gttacaatga catcccgatg   1440

tggaactata gcaaactgcc ggaatccttt ggcgcgacag aggatcgcgt ggtgagtaaa   1500

attgtgcgta cggaaaacga atttgtgtcg gttatgaaag aagcgcaggc tgacccgaat   1560

cgcatgtatt ggattgaact gatcctggca aaagaaggcg caccgaaagt tctgaaaaag   1620

atggggaaac tgtttgcgga gcaaaataaa agctaaggat cc                      1662


<210>  32
<211>  28
<212>  DNA
<213>  artificial sequence

<220>
<223>  primer

<400>  32
ggatccgcat gcttgcattt agtcgtgc                                        28


<210>  33
<211>  20
<212>  DNA
<213>  artificial sequence

<220>
<223>  primer

<400>  33
gggatgcgga cgtattcggc                                                 20


<210>  34
<211>  39
<212>  DNA
<213>  artificial sequence

<220>
<223>  primer

<400>  34
cacacatatt acaatagcta gctgaggatg aaagctctg                            39


<210>  35
<211>  39
<212>  DNA
<213>  artificial sequence

<220>
<223>  primer

<400>  35
cagagctttc atcctcagct agctattgta atatgtgtg                            39


