                         SEQUENCE LISTING

<110>  Evonik Industries AG
 
<120>  ACYL AMINO ACID PRODUCTION

<130>  201400319EP

<150>  15163499.5
<151>  2015-04-14

<160>  21    

<170>  PatentIn version 3.5

<210>  1
<211>  561
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  FadD Acyl-CoA synthase AA

<400>  1

Met Lys Lys Val Trp Leu Asn Arg Tyr Pro Ala Asp Val Pro Thr Glu 
1               5                   10                  15      


Ile Asn Pro Asp Arg Tyr Gln Ser Leu Val Asp Met Phe Glu Gln Ser 
            20                  25                  30          


Val Ala Arg Tyr Ala Asp Gln Pro Ala Phe Val Asn Met Gly Glu Val 
        35                  40                  45              


Met Thr Phe Arg Lys Leu Glu Glu Arg Ser Arg Ala Phe Ala Ala Tyr 
    50                  55                  60                  


Leu Gln Gln Gly Leu Gly Leu Lys Lys Gly Asp Arg Val Ala Leu Met 
65                  70                  75                  80  


Met Pro Asn Leu Leu Gln Tyr Pro Val Ala Leu Phe Gly Ile Leu Arg 
                85                  90                  95      


Ala Gly Met Ile Val Val Asn Val Asn Pro Leu Tyr Thr Pro Arg Glu 
            100                 105                 110         


Leu Glu His Gln Leu Asn Asp Ser Gly Ala Ser Ala Ile Val Ile Val 
        115                 120                 125             


Ser Asn Phe Ala His Thr Leu Glu Lys Val Val Asp Lys Thr Ala Val 
    130                 135                 140                 


Gln His Val Ile Leu Thr Arg Met Gly Asp Gln Leu Ser Thr Ala Lys 
145                 150                 155                 160 


Gly Thr Val Val Asn Phe Val Val Lys Tyr Ile Lys Arg Leu Val Pro 
                165                 170                 175     


Lys Tyr His Leu Pro Asp Ala Ile Ser Phe Arg Ser Ala Leu His Asn 
            180                 185                 190         


Gly Tyr Arg Met Gln Tyr Val Lys Pro Glu Leu Val Pro Glu Asp Leu 
        195                 200                 205             


Ala Phe Leu Gln Tyr Thr Gly Gly Thr Thr Gly Val Ala Lys Gly Ala 
    210                 215                 220                 


Met Leu Thr His Arg Asn Met Leu Ala Asn Leu Glu Gln Val Asn Ala 
225                 230                 235                 240 


Thr Tyr Gly Pro Leu Leu His Pro Gly Lys Glu Leu Val Val Thr Ala 
                245                 250                 255     


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


Ile Glu Leu Gly Gly Gln Asn Leu Leu Ile Thr Asn Pro Arg Asp Ile 
        275                 280                 285             


Pro Gly Leu Val Lys Glu Leu Ala Lys Tyr Pro Phe Thr Ala Ile Thr 
    290                 295                 300                 


Gly Val Asn Thr Leu Phe Asn Ala Leu Leu Asn Asn Lys Glu Phe Gln 
305                 310                 315                 320 


Gln Leu Asp Phe Ser Ser Leu His Leu Ser Ala Gly Gly Gly Met Pro 
                325                 330                 335     


Val Gln Gln Val Val Ala Glu Arg Trp Val Lys Leu Thr Gly Gln Tyr 
            340                 345                 350         


Leu Leu Glu Gly Tyr Gly Leu Thr Glu Cys Ala Pro Leu Val Ser Val 
        355                 360                 365             


Asn Pro Tyr Asp Ile Asp Tyr His Ser Gly Ser Ile Gly Leu Pro Val 
    370                 375                 380                 


Pro Ser Thr Glu Ala Lys Leu Val Asp Asp Asp Asp Asn Glu Val Pro 
385                 390                 395                 400 


Pro Gly Gln Pro Gly Glu Leu Cys Val Lys Gly Pro Gln Val Met Leu 
                405                 410                 415     


Gly Tyr Trp Gln Arg Pro Asp Ala Thr Asp Glu Ile Ile Lys Asn Gly 
            420                 425                 430         


Trp Leu His Thr Gly Asp Ile Ala Val Met Asp Glu Glu Gly Phe Leu 
        435                 440                 445             


Arg Ile Val Asp Arg Lys Lys Asp Met Ile Leu Val Ser Gly Phe Asn 
    450                 455                 460                 


Val Tyr Pro Asn Glu Ile Glu Asp Val Val Met Gln His Pro Gly Val 
465                 470                 475                 480 


Gln Glu Val Ala Ala Val Gly Val Pro Ser Gly Ser Ser Gly Glu Ala 
                485                 490                 495     


Val Lys Ile Phe Val Val Lys Lys Asp Pro Ser Leu Thr Glu Glu Ser 
            500                 505                 510         


Leu Val Thr Phe Cys Arg Arg Gln Leu Thr Gly Tyr Lys Val Pro Lys 
        515                 520                 525             


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


Leu Arg Arg Glu Leu Arg Asp Glu Ala Arg Gly Lys Val Asp Asn Lys 
545                 550                 555                 560 


Ala 
    


<210>  2
<211>  1680
<212>  DNA
<213>  Artificial Sequence

<220>
<223>   fadD (an acyl-CoA synthetase)

<400>  2
ttgaagaagg tttggcttaa ccgttatccc gcggacgttc cgacggagat caaccctgac       60

cgttatcaat ctctggtaga tatgtttgag cagtcggtcg cgcgctacgc cgatcaacct      120

gcgtttgtga atatggggga ggtaatgacc ttccgcaagc tggaagaacg cagtcgcgcg      180

tttgccgctt atttgcaaca agggttgggg ctgaagaaag gcgatcgcgt tgcgttgatg      240

atgcctaatt tattgcaata tccggtggcg ctgtttggca ttttgcgtgc cgggatgatc      300

gtcgtaaacg ttaacccgtt gtataccccg cgtgagcttg agcatcagct taacgatagc      360

ggcgcatcgg cgattgttat cgtgtctaac tttgctcaca cactggaaaa agtggttgat      420

aaaaccgccg ttcagcacgt aattctgacc cgtatgggcg atcagctatc tacggcaaaa      480

ggcacggtag tcaatttcgt tgttaaatac atcaagcgtt tggtgccgaa ataccatctg      540

ccagatgcca tttcatttcg tagcgcactg cataacggct accggatgca gtacgtcaaa      600

cccgaactgg tgccggaaga tttagctttt ctgcaataca ccggcggcac cactggtgtg      660

gcgaaaggcg cgatgctgac tcaccgcaat atgctggcga acctggaaca ggttaacgcg      720

acctatggtc cgctgttgca tccgggcaaa gagctggtgg tgacggcgct gccgctgtat      780

cacatttttg ccctgaccat taactgcctg ctgtttatcg aactgggtgg gcagaacctg      840

cttatcacta acccgcgcga tattccaggg ttggtaaaag agttagcgaa atatccgttt      900

accgctatca cgggcgttaa caccttgttc aatgcgttgc tgaacaataa agagttccag      960

cagctggatt tctccagtct gcatctttcc gcaggcggtg ggatgccagt gcagcaagtg     1020

gtggcagagc gttgggtgaa actgaccgga cagtatctgc tggaaggcta tggccttacc     1080

gagtgtgcgc cgctggtcag cgttaaccca tatgatattg attatcatag tggtagcatc     1140

ggtttgccgg tgccgtcgac ggaagccaaa ctggtggatg atgatgataa tgaagtacca     1200

ccaggtcaac cgggtgagct ttgtgtcaaa ggaccgcagg tgatgctggg ttactggcag     1260

cgtcccgatg ctaccgatga aatcatcaaa aatggctggt tacacaccgg cgacatcgcg     1320

gtaatggatg aagaaggatt cctgcgcatt gtcgatcgta aaaaagacat gattctggtt     1380

tccggtttta acgtctatcc caacgagatt gaagatgtcg tcatgcagca tcctggcgta     1440

caggaagtcg cggctgttgg cgtaccttcc ggctccagtg gtgaagcggt gaaaatcttc     1500

gtagtgaaaa aagatccatc gcttaccgaa gagtcactgg tgactttttg ccgccgtcag     1560

ctcacgggat acaaagtacc gaagctggtg gagtttcgtg atgagttacc gaaatctaac     1620

gtcggaaaaa ttttgcgacg agaattacgt gacgaagcgc gcggcaaagt ggacaataaa     1680


<210>  3
<211>  301
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  synUcTE (an acyl-CoA thioesterase) gene (codon-optimized)

<400>  3

Met Thr Leu Glu Trp Lys Pro Lys Pro Lys Leu Pro Gln Leu Leu Asp 
1               5                   10                  15      


Asp His Phe Gly Leu His Gly Leu Val Phe Arg Arg Thr Phe Ala Ile 
            20                  25                  30          


Arg Ser Tyr Glu Val Gly Pro Asp Arg Ser Thr Ser Ile Leu Ala Val 
        35                  40                  45              


Met Asn His Met Gln Glu Ala Thr Leu Asn His Ala Lys Ser Val Gly 
    50                  55                  60                  


Ile Leu Gly Asp Gly Phe Gly Thr Thr Leu Glu Met Ser Lys Arg Asp 
65                  70                  75                  80  


Leu Met Trp Val Val Arg Arg Thr His Val Ala Val Glu Arg Tyr Pro 
                85                  90                  95      


Thr Trp Gly Asp Thr Val Glu Val Glu Cys Trp Ile Gly Ala Ser Gly 
            100                 105                 110         


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


Glu Ile Leu Thr Arg Cys Thr Ser Leu Ser Val Leu Met Asn Thr Arg 
    130                 135                 140                 


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


Pro Ala Phe Ile Asp Asn Val Ala Val Lys Asp Asp Glu Ile Lys Lys 
                165                 170                 175     


Leu Gln Lys Leu Asn Asp Ser Thr Ala Asp Tyr Ile Gln Gly Gly Leu 
            180                 185                 190         


Thr Pro Arg Trp Asn Asp Leu Asp Val Asn Gln His Val Asn Asn Leu 
        195                 200                 205             


Lys Tyr Val Ala Trp Val Phe Glu Thr Val Pro Asp Ser Ile Phe Glu 
    210                 215                 220                 


Ser His His Ile Ser Ser Phe Thr Leu Glu Tyr Arg Arg Glu Cys Thr 
225                 230                 235                 240 


Arg Asp Ser Val Leu Arg Ser Leu Thr Thr Val Ser Gly Gly Ser Ser 
                245                 250                 255     


Glu Ala Gly Leu Val Cys Asp His Leu Leu Gln Leu Glu Gly Gly Ser 
            260                 265                 270         


Glu Val Leu Arg Ala Arg Thr Glu Trp Arg Pro Lys Leu Thr Asp Ser 
        275                 280                 285             


Phe Arg Gly Ile Ser Val Ile Pro Ala Glu Pro Arg Val 
    290                 295                 300     


<210>  4
<211>  288
<212>  PRT
<213>  Homo sapiens

<400>  4

Met Leu Val Leu Asn Cys Ser Thr Lys Leu Leu Ile Leu Glu Lys Met 
1               5                   10                  15      


Leu Lys Ser Cys Phe Pro Glu Ser Leu Lys Val Tyr Gly Ala Val Met 
            20                  25                  30          


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


Trp Pro Asp Phe Lys Ala Val Ile Thr Arg Arg Gln Arg Glu Ala Glu 
    50                  55                  60                  


Thr Asp Asn Leu Asp His Tyr Thr Asn Ala Tyr Ala Val Phe Tyr Lys 
65                  70                  75                  80  


Asp Val Arg Ala Tyr Arg Gln Leu Leu Glu Glu Cys Asp Val Phe Asn 
                85                  90                  95      


Trp Asp Gln Val Phe Gln Ile Gln Gly Leu Gln Ser Glu Leu Tyr Asp 
            100                 105                 110         


Val Ser Lys Ala Val Ala Asn Ser Lys Gln Leu Asn Ile Lys Leu Thr 
        115                 120                 125             


Ser Phe Lys Ala Val His Phe Ser Pro Val Ser Ser Leu Pro Asp Thr 
    130                 135                 140                 


Ser Phe Leu Lys Gly Pro Ser Pro Arg Leu Thr Tyr Leu Ser Val Ala 
145                 150                 155                 160 


Asn Ala Asp Leu Leu Asn Arg Thr Trp Ser Arg Gly Gly Asn Glu Gln 
                165                 170                 175     


Cys Leu Arg Tyr Ile Ala Asn Leu Ile Ser Cys Phe Pro Ser Val Cys 
            180                 185                 190         


Val Arg Asp Glu Lys Gly Asn Pro Val Ser Trp Ser Ile Thr Asp Gln 
        195                 200                 205             


Phe Ala Thr Met Cys His Gly Tyr Thr Leu Pro Glu His Arg Arg Lys 
    210                 215                 220                 


Gly Tyr Ser Arg Leu Val Ala Leu Thr Leu Ala Arg Lys Leu Gln Ser 
225                 230                 235                 240 


Arg Gly Phe Pro Ser Gln Gly Asn Val Leu Asp Asp Asn Thr Ala Ser 
                245                 250                 255     


Ile Ser Leu Leu Lys Ser Leu His Ala Glu Phe Leu Pro Cys Arg Phe 
            260                 265                 270         


His Arg Leu Ile Leu Thr Pro Ala Thr Phe Ser Gly Leu Pro His Leu 
        275                 280                 285             


<210>  5
<211>  867
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  glycine N-acyltransferase

<400>  5
atgttggtgc taaactgttc taccaaatta ctgatactgg agaaaatgtt gaagagttgc       60

tttcctgaat cactcaaggt ttacggagcg gtgatgaaca taaatcgtgg gaaccccttt      120

caaaaggaag tggtgttgga ttcatggccg gatttcaaag ctgttatcac ccgacgacaa      180

agagaggctg agacagataa ccttgatcat tatactaatg cctatgctgt gttctacaag      240

gatgtcaggg cttatcgaca gctattggaa gaatgtgatg tttttaactg ggaccaagtt      300

tttcaaatac aagggctgca gagtgagtta tatgatgttt ccaaagcggt tgccaattca      360

aagcagttga atataaagct aacttccttc aaggctgttc atttttctcc tgtttcatct      420

ctgccagata ccagtttcct caaggggcct tccccacgac taacctacct gagtgttgcc      480

aatgcggatc tactcaaccg gacttggtcc cggggaggca atgaacaatg tctccggtac      540

atcgccaacc tcatctcctg cttccctagt gtgtgtgtcc gggatgagaa gggaaacccg      600

gtctcctggt ccatcacaga ccagtttgcc accatgtgcc atggctacac cctgccagaa      660

catcgcagga aaggttacag ccggctggtg gccctcacgc tggccaggaa gttgcaaagc      720

cggggattcc cctctcaggg gaacgtcctg gatgacaaca cggcgtctat aagcctcctg      780

aagagtctcc atgctgagtt cttgccttgt cgcttccaca ggcttattct cacccctgcg      840

actttctctg gcctgcctca cctctag                                          867


<210>  6
<211>  230
<212>  PRT
<213>  Pseudomonas putida

<400>  6

Met Ser Phe Ser Asn Tyr Lys Val Ile Ala Met Pro Val Leu Val Ala 
1               5                   10                  15      


Asn Phe Val Leu Gly Ala Ala Thr Ala Trp Ala Asn Glu Asn Tyr Pro 
            20                  25                  30          


Ala Lys Ser Ala Gly Tyr Asn Gln Gly Asp Trp Val Ala Ser Phe Asn 
        35                  40                  45              


Phe Ser Lys Val Tyr Val Gly Glu Glu Leu Gly Asp Leu Asn Val Gly 
    50                  55                  60                  


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


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


Phe Phe Val Gly Val Pro Ala Arg Ala Lys Phe Gln Gly Glu Lys Ser 
            100                 105                 110         


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


Leu Ser Leu Gln Tyr His Tyr Asp Ser Phe Glu Arg Leu Tyr Pro Tyr 
    130                 135                 140                 


Val Gly Val Gly Val Gly Arg Val Leu Phe Phe Asp Lys Thr Asp Gly 
145                 150                 155                 160 


Ala Leu Ser Ser Phe Asp Ile Lys Asp Lys Trp Ala Pro Ala Phe Gln 
                165                 170                 175     


Val Gly Leu Arg Tyr Asp Leu Gly Asn Ser Trp Met Leu Asn Ser Asp 
            180                 185                 190         


Val Arg Tyr Ile Pro Phe Lys Thr Asp Val Thr Gly Thr Leu Gly Pro 
        195                 200                 205             


Val Pro Val Ser Thr Lys Ile Glu Val Asp Pro Phe Ile Leu Ser Leu 
    210                 215                 220                 


Gly Ala Ser Tyr Val Phe 
225                 230 


<210>  7
<211>  230
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  alkL (mod 1)

<400>  7

Val Ser Phe Ser Asn Tyr Lys Val Ile Ala Met Pro Val Leu Val Ala 
1               5                   10                  15      


Asn Phe Val Leu Gly Ala Ala Thr Ala Trp Ala Asn Glu Asn Tyr Pro 
            20                  25                  30          


Ala Lys Ser Ala Gly Tyr Asn Gln Gly Asp Trp Val Ala Ser Phe Asn 
        35                  40                  45              


Phe Ser Lys Val Tyr Val Gly Glu Glu Leu Gly Asp Leu Asn Val Gly 
    50                  55                  60                  


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


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


Phe Phe Val Gly Val Pro Ala Arg Ala Lys Phe Gln Gly Glu Lys Ser 
            100                 105                 110         


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


Leu Ser Leu Gln Tyr His Tyr Asp Ser Phe Glu Arg Leu Tyr Pro Tyr 
    130                 135                 140                 


Val Gly Val Gly Val Gly Arg Val Leu Phe Phe Asp Lys Thr Asp Gly 
145                 150                 155                 160 


Ala Leu Ser Ser Phe Asp Ile Lys Asp Lys Trp Ala Pro Ala Phe Gln 
                165                 170                 175     


Val Gly Leu Arg Tyr Asp Leu Gly Asn Ser Trp Met Leu Asn Ser Asp 
            180                 185                 190         


Val Arg Tyr Ile Pro Phe Lys Thr Asp Val Thr Gly Thr Leu Gly Pro 
        195                 200                 205             


Val Pro Val Ser Thr Lys Ile Glu Val Asp Pro Phe Ile Leu Ser Leu 
    210                 215                 220                 


Gly Ala Ser Tyr Val Phe 
225                 230 


<210>  8
<211>  446
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  FadL

<400>  8

Met Ser Gln Lys Thr Leu Phe Thr Lys Ser Ala Leu Ala Val Ala Val 
1               5                   10                  15      


Ala Leu Ile Ser Thr Gln Ala Trp Ser Ala Gly Phe Gln Leu Asn Glu 
            20                  25                  30          


Phe Ser Ser Ser Gly Leu Gly Arg Ala Tyr Ser Gly Glu Gly Ala Ile 
        35                  40                  45              


Ala Asp Asp Ala Gly Asn Val Ser Arg Asn Pro Ala Leu Ile Thr Met 
    50                  55                  60                  


Phe Asp Arg Pro Thr Phe Ser Ala Gly Ala Val Tyr Ile Asp Pro Asp 
65                  70                  75                  80  


Val Asn Ile Ser Gly Thr Ser Pro Ser Gly Arg Ser Leu Lys Ala Asp 
                85                  90                  95      


Asn Ile Ala Pro Thr Ala Trp Val Pro Asn Met His Phe Val Ala Pro 
            100                 105                 110         


Ile Asn Asp Gln Phe Gly Trp Gly Ala Ser Ile Thr Ser Asn Tyr Gly 
        115                 120                 125             


Leu Ala Thr Glu Phe Asn Asp Thr Tyr Ala Gly Gly Ser Val Gly Gly 
    130                 135                 140                 


Thr Thr Asp Leu Glu Thr Met Asn Leu Asn Leu Ser Gly Ala Tyr Arg 
145                 150                 155                 160 


Leu Asn Asn Ala Trp Ser Phe Gly Leu Gly Phe Asn Ala Val Tyr Ala 
                165                 170                 175     


Arg Ala Lys Ile Glu Arg Phe Ala Gly Asp Leu Gly Gln Leu Val Ala 
            180                 185                 190         


Gly Gln Ile Met Gln Ser Pro Ala Gly Gln Thr Gln Gln Gly Gln Ala 
        195                 200                 205             


Leu Ala Ala Thr Ala Asn Gly Ile Asp Ser Asn Thr Lys Ile Ala His 
    210                 215                 220                 


Leu Asn Gly Asn Gln Trp Gly Phe Gly Trp Asn Ala Gly Ile Leu Tyr 
225                 230                 235                 240 


Glu Leu Asp Lys Asn Asn Arg Tyr Ala Leu Thr Tyr Arg Ser Glu Val 
                245                 250                 255     


Lys Ile Asp Phe Lys Gly Asn Tyr Ser Ser Asp Leu Asn Arg Ala Phe 
            260                 265                 270         


Asn Asn Tyr Gly Leu Pro Ile Pro Thr Ala Thr Gly Gly Ala Thr Gln 
        275                 280                 285             


Ser Gly Tyr Leu Thr Leu Asn Leu Pro Glu Met Trp Glu Val Ser Gly 
    290                 295                 300                 


Tyr Asn Arg Val Asp Pro Gln Trp Ala Ile His Tyr Ser Leu Ala Tyr 
305                 310                 315                 320 


Thr Ser Trp Ser Gln Phe Gln Gln Leu Lys Ala Thr Ser Thr Ser Gly 
                325                 330                 335     


Asp Thr Leu Phe Gln Lys His Glu Gly Phe Lys Asp Ala Tyr Arg Ile 
            340                 345                 350         


Ala Leu Gly Thr Thr Tyr Tyr Tyr Asp Asp Asn Trp Thr Phe Arg Thr 
        355                 360                 365             


Gly Ile Ala Phe Asp Asp Ser Pro Val Pro Ala Gln Asn Arg Ser Ile 
    370                 375                 380                 


Ser Ile Pro Asp Gln Asp Arg Phe Trp Leu Ser Ala Gly Thr Thr Tyr 
385                 390                 395                 400 


Ala Phe Asn Lys Asp Ala Ser Val Asp Val Gly Val Ser Tyr Met His 
                405                 410                 415     


Gly Gln Ser Val Lys Ile Asn Glu Gly Pro Tyr Gln Phe Glu Ser Glu 
            420                 425                 430         


Gly Lys Ala Trp Leu Phe Gly Thr Asn Phe Asn Tyr Ala Phe 
        435                 440                 445     


<210>  9
<211>  39
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  gdhA_Up_fw

<400>  9
tcggtacccg gggatcgcct ttacgcgccg ccagaccag                              39


<210>  10
<211>  31
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  primer sequence (gdhA_Up_rev)

<400>  10
catagatata aaacccttat atattaatac g                                      31


<210>  11
<211>  47
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  primer sequence (gdhA-DOWN_fw)

<400>  11
ataagggttt tatatctatg gctcgagggg cgcagggtgt gatttaa                     47


<210>  12
<211>  38
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  primer sequence (gdhA-DOWN_rev)

<400>  12
tcgactctag aggatcgcgc taatcatagc tccggtaa                               38


<210>  13
<211>  523
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PCR result (gdhA_UP gdhA)

<400>  13
tcggtacccg gggatcgcct ttacgcgccg ccagaccagc ggattagtgc taagggtttt       60

gtcatcacgc tggcattgca gcagtattcc ttcggcttta attaccgccc cttcagaata      120

attttgatcc tgataaacgc agcactgagt acagggctgc gctgactgcc cgcctgaact      180

gaacacttct ggcggtacgt ttacctccac gtccggacga taatgcgggt tagccagtgc      240

aattaatgga aatgctaata ctacggcgaa caatgctcga ctcacaggga actccttaac      300

gttattgtct ctgctactga taacggtagc cgggtggcaa aactttagcg tctgagttat      360

cgcatttggt tatgagatta ctctcgttat taatttgctt tcctgggtca tttttttctt      420

gcttaccgtc acattcttga tggtatagtc gaaaactgca aaagcacatg acataaacaa      480

cataagcaca atcgtattaa tatataaggg ttttatatct atg                        523


<210>  14
<211>  544
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PCR result (gdh_DOWN gdhA)

<400>  14
ataagggttt tatatctatg gctcgagggg cgcagggtgt gatttaagtt gtaaatgcct       60

gatggcgcta cgcttatcag gcctacaaat gggcacaatt cattgcagtt acgctctaat      120

gtaggccggg caagcgcagc gcccccggca aaatttcagg cgtttatgag tatttaacgg      180

atgatgctcc ccacggaaca tttcttatgg gccaacggca tttcttactg tagtgctccc      240

aaaactgctt gtcgtaacga taacacgctt caagttcagc atccgttaac tttctgcgat      300

agcagcagat atgccagtaa agaaatccca tttgactatt tttttgataa tcttcttcgc      360

tttcgaacaa ctcgtgcgcc tttcgagaag caagcattat ataatgccag gccagttctt      420

cttcaattgt cccgttttga aaagctgtgc ttgatatcga gatcatccat gataattccg      480

ccgcccatat tagcttcgcc gaggatttac cggagctatg attagcgcga tcctctagag      540

tcga                                                                   544


<210>  15
<211>  5667
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  vector pKO3

<400>  15
cctttcgtct tcgaataaat acctgtgacg gaagatcact tcgcagaata aataaatcct       60

ggtgtccctg ttgataccgg gaagccctgg gccaactttt ggcgaaaatg agacgttgat      120

cggcacgtaa gaggttccaa ctttcaccat aatgaaataa gatcactacc gggcgtattt      180

tttgagttat cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga      240

tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca      300

gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc      360

gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg      420

aatgctcatc cggaattccg tatggcaatg aaagacggtg agctggtgat atgggatagt      480

gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt      540

gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac      600

ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt cgtctcagcc      660

aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc      720

gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg      780

gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa      840

ttacaacagt actgcgatga gtggcagggc ggggcgtaat ttttttaagg cagttattgg      900

tgcccttaaa cgcctggttg ctacgcctga ataagtgata ataagcggat gaatggcaga      960

aattcgaaag caaattcgac ccggtcgtcg gttcagggca gggtcgttaa atagccgctt     1020

atgtctattg ctggtctcgg tacccgggga tcgcggccgc ggaccggatc ctctagagcg     1080

gccgcgatcc tctagagtcg accggtggcg aatgggacgc gccctgtagc ggcgcattaa     1140

gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc     1200

ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag     1260

ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca     1320

aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc     1380

gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa     1440

cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct     1500

attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa     1560

cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt     1620

tttctaaata cattcaaata tgtatccgct caccgcgatc ctttttaacc catcacatat     1680

acctgccgtt cactattatt tagtgaaatg agatattatg atattttctg aattgtgatt     1740

aaaaaggcaa ctttatgccc atgcaacaga aactataaaa aatacagaga atgaaaagaa     1800

acagatagat tttttagttc tttaggcccg tagtctgcaa atccttttat gattttctat     1860

caaacaaaag aggaaaatag accagttgca atccaaacga gagtctaata gaatgaggtc     1920

gaaaagtaaa tcgcgcgggt ttgttactga taaagcaggc aagacctaaa atgtgtaaag     1980

ggcaaagtgt atactttggc gtcacccctt acatatttta ggtctttttt tattgtgcgt     2040

aactaacttg ccatcttcaa acaggagggc tggaagaagc agaccgctaa cacagtacat     2100

aaaaaaggag acatgaacga tgaacatcaa aaagtttgca aaacaagcaa cagtattaac     2160

ctttactacc gcactgctgg caggaggcgc aactcaagcg tttgcgaaag aaacgaacca     2220

aaagccatat aaggaaacat acggcatttc ccatattaca cgccatgata tgctgcaaat     2280

ccctgaacag caaaaaaatg aaaaatatca agttcctgag ttcgattcgt ccacaattaa     2340

aaatatctct tctgcaaaag gcctggacgt ttgggacagc tggccattac aaaacgctga     2400

cggcactgtc gcaaactatc acggctacca catcgtcttt gcattagccg gagatcctaa     2460

aaatgcggat gacacatcga tttacatgtt ctatcaaaaa gtcggcgaaa cttctattga     2520

cagctggaaa aacgctggcc gcgtctttaa agacagcgac aaattcgatg caaatgattc     2580

tatcctaaaa gaccaaacac aagaatggtc aggttcagcc acatttacat ctgacggaaa     2640

aatccgttta ttctacactg atttctccgg taaacattac ggcaaacaaa cactgacaac     2700

tgcacaagtt aacgtatcag catcagacag ctctttgaac atcaacggtg tagaggatta     2760

taaatcaatc tttgacggtg acggaaaaac gtatcaaaat gtacagcagt tcatcgatga     2820

aggcaactac agctcaggcg acaaccatac gctgagagat cctcactacg tagaagataa     2880

aggccacaaa tacttagtat ttgaagcaaa cactggaact gaagatggct accaaggcga     2940

agaatcttta tttaacaaag catactatgg caaaagcaca tcattcttcc gtcaagaaag     3000

tcaaaaactt ctgcaaagcg ataaaaaacg cacggctgag ttagcaaacg gcgctctcgg     3060

tatgattgag ctaaacgatg attacacact gaaaaaagtg atgaaaccgc tgattgcatc     3120

taacacagta acagatgaaa ttgaacgcgc gaacgtcttt aaaatgaacg gcaaatggta     3180

cctgttcact gactcccgcg gatcaaaaat gacgattgac ggcattacgt ctaacgatat     3240

ttacatgctt ggttatgttt ctaattcttt aactggccca tacaagccgc tgaacaaaac     3300

tggccttgtg ttaaaaatgg atcttgatcc taacgatgta acctttactt actcacactt     3360

cgctgtacct caagcgaaag gaaacaatgt cgtgattaca agctatatga caaacagagg     3420

attctacgca gacaaacaat caacgtttgc gccaagcttc ctgctgaaca tcaaaggcaa     3480

gaaaacatct gttgtcaaag acagcatcct tgaacaagga caattaacag ttaacaaata     3540

aaaacgcaaa agaaaatgcc gatattgact accggaagca gtgtgaccgt gtgcttctca     3600

aatgcctgat tcaggctgtc tatgtgtgac tgttgagctg taacaagttg tctcaggtgt     3660

tcaatttcat gttctagttg ctttgtttta ctggtttcac ctgttctatt aggtgttaca     3720

tgctgttcat ctgttacatt gtcgatctgt tcatggtgaa cagctttaaa tgcaccaaaa     3780

actcgtaaaa gctctgatgt atctatcttt tttacaccgt tttcatctgt gcatatggac     3840

agttttccct ttgatatgta acggtgaaca gttgttctac ttttgtttgt tagtcttgat     3900

gcttcactga tagatacaag agccataaga acctcagatc cttccgtatt tagccagtat     3960

gttctctagt gtggttcgtt gtttttgcgt gagccatgag aacgaaccat tgagatcata     4020

cttactttgc atgtcactca aaaattttgc ctcaaaactg gtgagctgaa tttttgcagt     4080

taaagcatcg tgtagtgttt ttcttagtcc gttatgtagg taggaatctg atgtaatggt     4140

tgttggtatt ttgtcaccat tcatttttat ctggttgttc tcaagttcgg ttacgagatc     4200

catttgtcta tctagttcaa cttggaaaat caacgtatca gtcgggcggc ctcgcttatc     4260

aaccaccaat ttcatattgc tgtaagtgtt taaatcttta cttattggtt tcaaaaccca     4320

ttggttaagc cttttaaact catggtagtt attttcaagc attaacatga acttaaattc     4380

atcaaggcta atctctatat ttgccttgtg agttttcttt tgtgttagtt cttttaataa     4440

ccactcataa atcctcatag agtatttgtt ttcaaaagac ttaacatgtt ccagattata     4500

ttttatgaat ttttttaact ggaaaagata aggcaatatc tcttcactaa aaactaattc     4560

taatttttcg cttgagaact tggcatagtt tgtccactgg aaaatctcaa agcctttaac     4620

caaaggattc ctgatttcca cagttctcgt catcagctct ctggttgctt tagctaatac     4680

accataagca ttttccctac tgatgttcat catctgaacg tattggttat aagtgaacga     4740

taccgtccgt tctttccttg tagggttttc aatcgtgggg ttgagtagtg ccacacagca     4800

taaaattagc ttggtttcat gctccgttaa gtcatagcga ctaatcgcta gttcatttgc     4860

tttgaaaaca actaattcag acatacatct caattggtct aggtgatttt aatcactata     4920

ccaattgaga tgggctagtc aatgataatt actagtcctt ttcctttgag ttgtgggtat     4980

ctgtaaattc tgctagacct ttgctggaaa acttgtaaat tctgctagac cctctgtaaa     5040

ttccgctaga cctttgtgtg ttttttttgt ttatattcaa gtggttataa tttatagaat     5100

aaagaaagaa taaaaaaaga taaaaagaat agatcccagc cctgtgtata actcactact     5160

ttagtcagtt ccgcagtatt acaaaaggat gtcgcaaacg ctgtttgctc ctctacaaaa     5220

cagaccttaa aaccctaaag gcttaagtag caccctcgca agctcgggca aatcgctgaa     5280

tattcctttt gtctccgacc atcaggcacc tgagtcgctg tctttttcgt gacattcagt     5340

tcgctgcgct cacggctctg gcagtgaatg ggggtaaatg gcactacagg cgccttttat     5400

ggattcatgc aaggaaacta cccataatac aagaaaagcc cgtcacgggc ttctcagggc     5460

gttttatggc gggtctgcta tgtggtgcta tctgactttt tgctgttcag cagttcctgc     5520

cctctgattt tccagtctga ccacttcgga ttatcccgtg acaggtcatt cagactggct     5580

aatgcaccca gtaaggcagc ggtatcatca acaggcttac ccgtcttact gtcggggatc     5640

gacgctctcc cttatgcgac tcctgca                                         5667


<210>  16
<211>  6649
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  pKO3_KO-gdhA

<400>  16
cctttcgtct tcgaataaat acctgtgacg gaagatcact tcgcagaata aataaatcct       60

ggtgtccctg ttgataccgg gaagccctgg gccaactttt ggcgaaaatg agacgttgat      120

cggcacgtaa gaggttccaa ctttcaccat aatgaaataa gatcactacc gggcgtattt      180

tttgagttat cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga      240

tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca      300

gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc      360

gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg      420

aatgctcatc cggaattccg tatggcaatg aaagacggtg agctggtgat atgggatagt      480

gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt      540

gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac      600

ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt cgtctcagcc      660

aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc      720

gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg      780

gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa      840

ttacaacagt actgcgatga gtggcagggc ggggcgtaat ttttttaagg cagttattgg      900

tgcccttaaa cgcctggttg ctacgcctga ataagtgata ataagcggat gaatggcaga      960

aattcgaaag caaattcgac ccggtcgtcg gttcagggca gggtcgttaa atagccgctt     1020

atgtctattg ctggtctcgg tacccgggga tcgcctttac gcgccgccag accagcggat     1080

tagtgctaag ggttttgtca tcacgctggc attgcagcag tattccttcg gctttaatta     1140

ccgccccttc agaataattt tgatcctgat aaacgcagca ctgagtacag ggctgcgctg     1200

actgcccgcc tgaactgaac acttctggcg gtacgtttac ctccacgtcc ggacgataat     1260

gcgggttagc cagtgcaatt aatggaaatg ctaatactac ggcgaacaat gctcgactca     1320

cagggaactc cttaacgtta ttgtctctgc tactgataac ggtagccggg tggcaaaact     1380

ttagcgtctg agttatcgca tttggttatg agattactct cgttattaat ttgctttcct     1440

gggtcatttt tttcttgctt accgtcacat tcttgatggt atagtcgaaa actgcaaaag     1500

cacatgacat aaacaacata agcacaatcg tattaatata taagggtttt atatctatgg     1560

ctcgaggggc gcagggtgtg atttaagttg taaatgcctg atggcgctac gcttatcagg     1620

cctacaaatg ggcacaattc attgcagtta cgctctaatg taggccgggc aagcgcagcg     1680

cccccggcaa aatttcaggc gtttatgagt atttaacgga tgatgctccc cacggaacat     1740

ttcttatggg ccaacggcat ttcttactgt agtgctccca aaactgcttg tcgtaacgat     1800

aacacgcttc aagttcagca tccgttaact ttctgcgata gcagcagata tgccagtaaa     1860

gaaatcccat ttgactattt ttttgataat cttcttcgct ttcgaacaac tcgtgcgcct     1920

ttcgagaagc aagcattata taatgccagg ccagttcttc ttcaattgtc ccgttttgaa     1980

aagctgtgct tgatatcgag atcatccatg ataattccgc cgcccatatt agcttcgccg     2040

aggatttacc ggagctatga ttagcgcgat cctctagagt cgaccggtgg cgaatgggac     2100

gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct     2160

acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg     2220

ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt     2280

gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca     2340

tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga     2400

ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa     2460

gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac     2520

gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc     2580

gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcaccgcga     2640

tcctttttaa cccatcacat atacctgccg ttcactatta tttagtgaaa tgagatatta     2700

tgatattttc tgaattgtga ttaaaaaggc aactttatgc ccatgcaaca gaaactataa     2760

aaaatacaga gaatgaaaag aaacagatag attttttagt tctttaggcc cgtagtctgc     2820

aaatcctttt atgattttct atcaaacaaa agaggaaaat agaccagttg caatccaaac     2880

gagagtctaa tagaatgagg tcgaaaagta aatcgcgcgg gtttgttact gataaagcag     2940

gcaagaccta aaatgtgtaa agggcaaagt gtatactttg gcgtcacccc ttacatattt     3000

taggtctttt tttattgtgc gtaactaact tgccatcttc aaacaggagg gctggaagaa     3060

gcagaccgct aacacagtac ataaaaaagg agacatgaac gatgaacatc aaaaagtttg     3120

caaaacaagc aacagtatta acctttacta ccgcactgct ggcaggaggc gcaactcaag     3180

cgtttgcgaa agaaacgaac caaaagccat ataaggaaac atacggcatt tcccatatta     3240

cacgccatga tatgctgcaa atccctgaac agcaaaaaaa tgaaaaatat caagttcctg     3300

agttcgattc gtccacaatt aaaaatatct cttctgcaaa aggcctggac gtttgggaca     3360

gctggccatt acaaaacgct gacggcactg tcgcaaacta tcacggctac cacatcgtct     3420

ttgcattagc cggagatcct aaaaatgcgg atgacacatc gatttacatg ttctatcaaa     3480

aagtcggcga aacttctatt gacagctgga aaaacgctgg ccgcgtcttt aaagacagcg     3540

acaaattcga tgcaaatgat tctatcctaa aagaccaaac acaagaatgg tcaggttcag     3600

ccacatttac atctgacgga aaaatccgtt tattctacac tgatttctcc ggtaaacatt     3660

acggcaaaca aacactgaca actgcacaag ttaacgtatc agcatcagac agctctttga     3720

acatcaacgg tgtagaggat tataaatcaa tctttgacgg tgacggaaaa acgtatcaaa     3780

atgtacagca gttcatcgat gaaggcaact acagctcagg cgacaaccat acgctgagag     3840

atcctcacta cgtagaagat aaaggccaca aatacttagt atttgaagca aacactggaa     3900

ctgaagatgg ctaccaaggc gaagaatctt tatttaacaa agcatactat ggcaaaagca     3960

catcattctt ccgtcaagaa agtcaaaaac ttctgcaaag cgataaaaaa cgcacggctg     4020

agttagcaaa cggcgctctc ggtatgattg agctaaacga tgattacaca ctgaaaaaag     4080

tgatgaaacc gctgattgca tctaacacag taacagatga aattgaacgc gcgaacgtct     4140

ttaaaatgaa cggcaaatgg tacctgttca ctgactcccg cggatcaaaa atgacgattg     4200

acggcattac gtctaacgat atttacatgc ttggttatgt ttctaattct ttaactggcc     4260

catacaagcc gctgaacaaa actggccttg tgttaaaaat ggatcttgat cctaacgatg     4320

taacctttac ttactcacac ttcgctgtac ctcaagcgaa aggaaacaat gtcgtgatta     4380

caagctatat gacaaacaga ggattctacg cagacaaaca atcaacgttt gcgccaagct     4440

tcctgctgaa catcaaaggc aagaaaacat ctgttgtcaa agacagcatc cttgaacaag     4500

gacaattaac agttaacaaa taaaaacgca aaagaaaatg ccgatattga ctaccggaag     4560

cagtgtgacc gtgtgcttct caaatgcctg attcaggctg tctatgtgtg actgttgagc     4620

tgtaacaagt tgtctcaggt gttcaatttc atgttctagt tgctttgttt tactggtttc     4680

acctgttcta ttaggtgtta catgctgttc atctgttaca ttgtcgatct gttcatggtg     4740

aacagcttta aatgcaccaa aaactcgtaa aagctctgat gtatctatct tttttacacc     4800

gttttcatct gtgcatatgg acagttttcc ctttgatatg taacggtgaa cagttgttct     4860

acttttgttt gttagtcttg atgcttcact gatagataca agagccataa gaacctcaga     4920

tccttccgta tttagccagt atgttctcta gtgtggttcg ttgtttttgc gtgagccatg     4980

agaacgaacc attgagatca tacttacttt gcatgtcact caaaaatttt gcctcaaaac     5040

tggtgagctg aatttttgca gttaaagcat cgtgtagtgt ttttcttagt ccgttatgta     5100

ggtaggaatc tgatgtaatg gttgttggta ttttgtcacc attcattttt atctggttgt     5160

tctcaagttc ggttacgaga tccatttgtc tatctagttc aacttggaaa atcaacgtat     5220

cagtcgggcg gcctcgctta tcaaccacca atttcatatt gctgtaagtg tttaaatctt     5280

tacttattgg tttcaaaacc cattggttaa gccttttaaa ctcatggtag ttattttcaa     5340

gcattaacat gaacttaaat tcatcaaggc taatctctat atttgccttg tgagttttct     5400

tttgtgttag ttcttttaat aaccactcat aaatcctcat agagtatttg ttttcaaaag     5460

acttaacatg ttccagatta tattttatga atttttttaa ctggaaaaga taaggcaata     5520

tctcttcact aaaaactaat tctaattttt cgcttgagaa cttggcatag tttgtccact     5580

ggaaaatctc aaagccttta accaaaggat tcctgatttc cacagttctc gtcatcagct     5640

ctctggttgc tttagctaat acaccataag cattttccct actgatgttc atcatctgaa     5700

cgtattggtt ataagtgaac gataccgtcc gttctttcct tgtagggttt tcaatcgtgg     5760

ggttgagtag tgccacacag cataaaatta gcttggtttc atgctccgtt aagtcatagc     5820

gactaatcgc tagttcattt gctttgaaaa caactaattc agacatacat ctcaattggt     5880

ctaggtgatt ttaatcacta taccaattga gatgggctag tcaatgataa ttactagtcc     5940

ttttcctttg agttgtgggt atctgtaaat tctgctagac ctttgctgga aaacttgtaa     6000

attctgctag accctctgta aattccgcta gacctttgtg tgtttttttt gtttatattc     6060

aagtggttat aatttataga ataaagaaag aataaaaaaa gataaaaaga atagatccca     6120

gccctgtgta taactcacta ctttagtcag ttccgcagta ttacaaaagg atgtcgcaaa     6180

cgctgtttgc tcctctacaa aacagacctt aaaaccctaa aggcttaagt agcaccctcg     6240

caagctcggg caaatcgctg aatattcctt ttgtctccga ccatcaggca cctgagtcgc     6300

tgtctttttc gtgacattca gttcgctgcg ctcacggctc tggcagtgaa tgggggtaaa     6360

tggcactaca ggcgcctttt atggattcat gcaaggaaac tacccataat acaagaaaag     6420

cccgtcacgg gcttctcagg gcgttttatg gcgggtctgc tatgtggtgc tatctgactt     6480

tttgctgttc agcagttcct gccctctgat tttccagtct gaccacttcg gattatcccg     6540

tgacaggtca ttcagactgg ctaatgcacc cagtaaggca gcggtatcat caacaggctt     6600

acccgtctta ctgtcgggga tcgacgctct cccttatgcg actcctgca                 6649


<210>  17
<211>  30
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  DNA sequence after deletion of gdhA

<400>  17
atggctcgag gggcgcaggg tgtgatttaa                                        30


<210>  18
<211>  447
<212>  PRT
<213>  Escherichia coli

<400>  18

Met Asp Gln Thr Tyr Ser Leu Glu Ser Phe Leu Asn His Val Gln Lys 
1               5                   10                  15      


Arg Asp Pro Asn Gln Thr Glu Phe Ala Gln Ala Val Arg Glu Val Met 
            20                  25                  30          


Thr Thr Leu Trp Pro Phe Leu Glu Gln Asn Pro Lys Tyr Arg Gln Met 
        35                  40                  45              


Ser Leu Leu Glu Arg Leu Val Glu Pro Glu Arg Val Ile Gln Phe Arg 
    50                  55                  60                  


Val Val Trp Val Asp Asp Arg Asn Gln Ile Gln Val Asn Arg Ala Trp 
65                  70                  75                  80  


Arg Val Gln Phe Ser Ser Ala Ile Gly Pro Tyr Lys Gly Gly Met Arg 
                85                  90                  95      


Phe His Pro Ser Val Asn Leu Ser Ile Leu Lys Phe Leu Gly Phe Glu 
            100                 105                 110         


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


Gly Gly Ser Asp Phe Asp Pro Lys Gly Lys Ser Glu Gly Glu Val Met 
    130                 135                 140                 


Arg Phe Cys Gln Ala Leu Met Thr Glu Leu Tyr Arg His Leu Gly Ala 
145                 150                 155                 160 


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


Gly Phe Met Ala Gly Met Met Lys Lys Leu Ser Asn Asn Thr Ala Cys 
            180                 185                 190         


Val Phe Thr Gly Lys Gly Leu Ser Phe Gly Gly Ser Leu Ile Arg Pro 
        195                 200                 205             


Glu Ala Thr Gly Tyr Gly Leu Val Tyr Phe Thr Glu Ala Met Leu Lys 
    210                 215                 220                 


Arg His Gly Met Gly Phe Glu Gly Met Arg Val Ser Val Ser Gly Ser 
225                 230                 235                 240 


Gly Asn Val Ala Gln Tyr Ala Ile Glu Lys Ala Met Glu Phe Gly Ala 
                245                 250                 255     


Arg Val Ile Thr Ala Ser Asp Ser Ser Gly Thr Val Val Asp Glu Ser 
            260                 265                 270         


Gly Phe Thr Lys Glu Lys Leu Ala Arg Leu Ile Glu Ile Lys Ala Ser 
        275                 280                 285             


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


Leu Glu Gly Gln Gln Pro Trp Ser Leu Pro Val Asp Ile Ala Leu Pro 
305                 310                 315                 320 


Cys Ala Thr Gln Asn Glu Leu Asp Val Asp Ala Ala His Gln Leu Ile 
                325                 330                 335     


Ala Asn Gly Val Lys Ala Val Ala Glu Gly Ala Asn Met Pro Thr Thr 
            340                 345                 350         


Ile Glu Ala Thr Glu Leu Phe Gln Gln Ala Gly Val Leu Phe Ala Pro 
        355                 360                 365             


Gly Lys Ala Ala Asn Ala Gly Gly Val Ala Thr Ser Gly Leu Glu Met 
    370                 375                 380                 


Ala Gln Asn Ala Ala Arg Leu Gly Trp Lys Ala Glu Lys Val Asp Ala 
385                 390                 395                 400 


Arg Leu His His Ile Met Leu Asp Ile His His Ala Cys Val Glu His 
                405                 410                 415     


Gly Gly Glu Gly Glu Gln Thr Asn Tyr Val Gln Gly Ala Asn Ile Ala 
            420                 425                 430         


Gly Phe Val Lys Val Ala Asp Ala Met Leu Ala Gln Gly Val Ile 
        435                 440                 445         


<210>  19
<211>  466
<212>  PRT
<213>  Escherichia coli

<400>  19

Met Asp Gln Lys Leu Leu Thr Asp Phe Arg Ser Glu Leu Leu Asp Ser 
1               5                   10                  15      


Arg Phe Gly Ala Lys Ala Ile Ser Thr Ile Ala Glu Ser Lys Arg Phe 
            20                  25                  30          


Pro Leu His Glu Met Arg Asp Asp Val Ala Phe Gln Ile Ile Asn Asp 
        35                  40                  45              


Glu Leu Tyr Leu Asp Gly Asn Ala Arg Gln Asn Leu Ala Thr Phe Cys 
    50                  55                  60                  


Gln Thr Trp Asp Asp Glu Asn Val His Lys Leu Met Asp Leu Ser Ile 
65                  70                  75                  80  


Asn Lys Asn Trp Ile Asp Lys Glu Glu Tyr Pro Gln Ser Ala Ala Ile 
                85                  90                  95      


Asp Leu Arg Cys Val Asn Met Val Ala Asp Leu Trp His Ala Pro Ala 
            100                 105                 110         


Pro Lys Asn Gly Gln Ala Val Gly Thr Asn Thr Ile Gly Ser Ser Glu 
        115                 120                 125             


Ala Cys Met Leu Gly Gly Met Ala Met Lys Trp Arg Trp Arg Lys Arg 
    130                 135                 140                 


Met Glu Ala Ala Gly Lys Pro Thr Asp Lys Pro Asn Leu Val Cys Gly 
145                 150                 155                 160 


Pro Val Gln Ile Cys Trp His Lys Phe Ala Arg Tyr Trp Asp Val Glu 
                165                 170                 175     


Leu Arg Glu Ile Pro Met Arg Pro Gly Gln Leu Phe Met Asp Pro Lys 
            180                 185                 190         


Arg Met Ile Glu Ala Cys Asp Glu Asn Thr Ile Gly Val Val Pro Thr 
        195                 200                 205             


Phe Gly Val Thr Tyr Thr Gly Asn Tyr Glu Phe Pro Gln Pro Leu His 
    210                 215                 220                 


Asp Ala Leu Asp Lys Phe Gln Ala Asp Thr Gly Ile Asp Ile Asp Met 
225                 230                 235                 240 


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


Asp Ile Val Trp Asp Phe Arg Leu Pro Arg Val Lys Ser Ile Ser Ala 
            260                 265                 270         


Ser Gly His Lys Phe Gly Leu Ala Pro Leu Gly Cys Gly Trp Val Ile 
        275                 280                 285             


Trp Arg Asp Glu Glu Ala Leu Pro Gln Glu Leu Val Phe Asn Val Asp 
    290                 295                 300                 


Tyr Leu Gly Gly Gln Ile Gly Thr Phe Ala Ile Asn Phe Ser Arg Pro 
305                 310                 315                 320 


Ala Gly Gln Val Ile Ala Gln Tyr Tyr Glu Phe Leu Arg Leu Gly Arg 
                325                 330                 335     


Glu Gly Tyr Thr Lys Val Gln Asn Ala Ser Tyr Gln Val Ala Ala Tyr 
            340                 345                 350         


Leu Ala Asp Glu Ile Ala Lys Leu Gly Pro Tyr Glu Phe Ile Cys Thr 
        355                 360                 365             


Gly Arg Pro Asp Glu Gly Ile Pro Ala Val Cys Phe Lys Leu Lys Asp 
    370                 375                 380                 


Gly Glu Asp Pro Gly Tyr Thr Leu Tyr Asp Leu Ser Glu Arg Leu Arg 
385                 390                 395                 400 


Leu Arg Gly Trp Gln Val Pro Ala Phe Thr Leu Gly Gly Glu Ala Thr 
                405                 410                 415     


Asp Ile Val Val Met Arg Ile Met Cys Arg Arg Gly Phe Glu Met Asp 
            420                 425                 430         


Phe Ala Glu Leu Leu Leu Glu Asp Tyr Lys Ala Ser Leu Lys Tyr Leu 
        435                 440                 445             


Ser Asp His Pro Lys Leu Gln Gly Ile Ala Gln Gln Asn Ser Phe Lys 
    450                 455                 460                 


His Thr 
465     


<210>  20
<211>  466
<212>  PRT
<213>  Escherichia coli

<400>  20

Met Asp Lys Lys Gln Val Thr Asp Leu Arg Ser Glu Leu Leu Asp Ser 
1               5                   10                  15      


Arg Phe Gly Ala Lys Ser Ile Ser Thr Ile Ala Glu Ser Lys Arg Phe 
            20                  25                  30          


Pro Leu His Glu Met Arg Asp Asp Val Ala Phe Gln Ile Ile Asn Asp 
        35                  40                  45              


Glu Leu Tyr Leu Asp Gly Asn Ala Arg Gln Asn Leu Ala Thr Phe Cys 
    50                  55                  60                  


Gln Thr Trp Asp Asp Glu Asn Val His Lys Leu Met Asp Leu Ser Ile 
65                  70                  75                  80  


Asn Lys Asn Trp Ile Asp Lys Glu Glu Tyr Pro Gln Ser Ala Ala Ile 
                85                  90                  95      


Asp Leu Arg Cys Val Asn Met Val Ala Asp Leu Trp His Ala Pro Ala 
            100                 105                 110         


Pro Lys Asn Gly Gln Ala Val Gly Thr Asn Thr Ile Gly Ser Ser Glu 
        115                 120                 125             


Ala Cys Met Leu Gly Gly Met Ala Met Lys Trp Arg Trp Arg Lys Arg 
    130                 135                 140                 


Met Glu Ala Ala Gly Lys Pro Thr Asp Lys Pro Asn Leu Val Cys Gly 
145                 150                 155                 160 


Pro Val Gln Ile Cys Trp His Lys Phe Ala Arg Tyr Trp Asp Val Glu 
                165                 170                 175     


Leu Arg Glu Ile Pro Met Arg Pro Gly Gln Leu Phe Met Asp Pro Lys 
            180                 185                 190         


Arg Met Ile Glu Ala Cys Asp Glu Asn Thr Ile Gly Val Val Pro Thr 
        195                 200                 205             


Phe Gly Val Thr Tyr Thr Gly Asn Tyr Glu Phe Pro Gln Pro Leu His 
    210                 215                 220                 


Asp Ala Leu Asp Lys Phe Gln Ala Asp Thr Gly Ile Asp Ile Asp Met 
225                 230                 235                 240 


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


Asp Ile Val Trp Asp Phe Arg Leu Pro Arg Val Lys Ser Ile Ser Ala 
            260                 265                 270         


Ser Gly His Lys Phe Gly Leu Ala Pro Leu Gly Cys Gly Trp Val Ile 
        275                 280                 285             


Trp Arg Asp Glu Glu Ala Leu Pro Gln Glu Leu Val Phe Asn Val Asp 
    290                 295                 300                 


Tyr Leu Gly Gly Gln Ile Gly Thr Phe Ala Ile Asn Phe Ser Arg Pro 
305                 310                 315                 320 


Ala Gly Gln Val Ile Ala Gln Tyr Tyr Glu Phe Leu Arg Leu Gly Arg 
                325                 330                 335     


Glu Gly Tyr Thr Lys Val Gln Asn Ala Ser Tyr Gln Val Ala Ala Tyr 
            340                 345                 350         


Leu Ala Asp Glu Ile Ala Lys Leu Gly Pro Tyr Glu Phe Ile Cys Thr 
        355                 360                 365             


Gly Arg Pro Asp Glu Gly Ile Pro Ala Val Cys Phe Lys Leu Lys Asp 
    370                 375                 380                 


Gly Glu Asp Pro Gly Tyr Thr Leu Tyr Asp Leu Ser Glu Arg Leu Arg 
385                 390                 395                 400 


Leu Arg Gly Trp Gln Val Pro Ala Phe Thr Leu Gly Gly Glu Ala Thr 
                405                 410                 415     


Asp Ile Val Val Met Arg Ile Met Cys Arg Arg Gly Phe Glu Met Asp 
            420                 425                 430         


Phe Ala Glu Leu Leu Leu Glu Asp Tyr Lys Ala Ser Leu Lys Tyr Leu 
        435                 440                 445             


Ser Asp His Pro Lys Leu Gln Gly Ile Ala Gln Gln Asn Ser Phe Lys 
    450                 455                 460                 


His Thr 
465     


<210>  21
<211>  396
<212>  PRT
<213>  Escherichia coli

<400>  21

Met Phe Glu Asn Ile Thr Ala Ala Pro Ala Asp Pro Ile Leu Gly Leu 
1               5                   10                  15      


Ala Asp Leu Phe Arg Ala Asp Glu Arg Pro Gly Lys Ile Asn Leu Gly 
            20                  25                  30          


Ile Gly Val Tyr Lys Asp Glu Thr Gly Lys Thr Pro Val Leu Thr Ser 
        35                  40                  45              


Val Lys Lys Ala Glu Gln Tyr Leu Leu Glu Asn Glu Thr Thr Lys Asn 
    50                  55                  60                  


Tyr Leu Gly Ile Asp Gly Ile Pro Glu Phe Gly Arg Cys Thr Gln Glu 
65                  70                  75                  80  


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


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


Phe Leu Ala Lys Asn Thr Ser Val Lys Arg Val Trp Val Ser Asn Pro 
        115                 120                 125             


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


Arg Glu Tyr Ala Tyr Tyr Asp Ala Glu Asn His Thr Leu Asp Phe Asp 
145                 150                 155                 160 


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


Phe His Gly Cys Cys His Asn Pro Thr Gly Ile Asp Pro Thr Leu Glu 
            180                 185                 190         


Gln Trp Gln Thr Leu Ala Gln Leu Ser Val Glu Lys Gly Trp Leu Pro 
        195                 200                 205             


Leu Phe Asp Phe Ala Tyr Gln Gly Phe Ala Arg Gly Leu Glu Glu Asp 
    210                 215                 220                 


Ala Glu Gly Leu Arg Ala Phe Ala Ala Met His Lys Glu Leu Ile Val 
225                 230                 235                 240 


Ala Ser Ser Tyr Ser Lys Asn Phe Gly Leu Tyr Asn Glu Arg Val Gly 
                245                 250                 255     


Ala Cys Thr Leu Val Ala Ala Asp Ser Glu Thr Val Asp Arg Ala Phe 
            260                 265                 270         


Ser Gln Met Lys Ala Ala Ile Arg Ala Asn Tyr Ser Asn Pro Pro Ala 
        275                 280                 285             


His Gly Ala Ser Val Val Ala Thr Ile Leu Ser Asn Asp Ala Leu Arg 
    290                 295                 300                 


Ala Ile Trp Glu Gln Glu Leu Thr Asp Met Arg Gln Arg Ile Gln Arg 
305                 310                 315                 320 


Met Arg Gln Leu Phe Val Asn Thr Leu Gln Glu Lys Gly Ala Asn Arg 
                325                 330                 335     


Asp Phe Ser Phe Ile Ile Lys Gln Asn Gly Met Phe Ser Phe Ser Gly 
            340                 345                 350         


Leu Thr Lys Glu Gln Val Leu Arg Leu Arg Glu Glu Phe Gly Val Tyr 
        355                 360                 365             


Ala Val Ala Ser Gly Arg Val Asn Val Ala Gly Met Thr Pro Asp Asn 
    370                 375                 380                 


Met Ala Pro Leu Cys Glu Ala Ile Val Ala Val Leu 
385                 390                 395     


