                         SEQUENCE LISTING

<110>  The Regents of the University of California
       Liao, James C
       Cho, Kwang Myung
 
<120>  PRODUCTION OF gamma-AMINOBUTYRIC ACID BY RECOMBINANT 
       MICROORGANISMS

<130>  UCLA136585

<150>  US 61/329204
<151>  2010-04-29

<160>  6     

<170>  PatentIn version 3.5

<210>  1
<211>  1401
<212>  DNA
<213>  Escherichia coli


<220>
<221>  CDS
<222>  (1)..(1401)

<400>  1
atg gac cag aag ctg tta acg gat ttc cgc tca gaa cta ctc gat tca         48
Met Asp Gln Lys Leu Leu Thr Asp Phe Arg Ser Glu Leu Leu Asp Ser           
1               5                   10                  15                

cgt ttt ggc gca aag gcc att tct act atc gcg gag tca aaa cga ttt         96
Arg Phe Gly Ala Lys Ala Ile Ser Thr Ile Ala Glu Ser Lys Arg Phe           
            20                  25                  30                    

ccg ctg cac gaa atg cgc gat gat gtc gca ttt cag att atc aat gat        144
Pro Leu His Glu Met Arg Asp Asp Val Ala Phe Gln Ile Ile Asn Asp           
        35                  40                  45                        

gaa tta tat ctt gat ggc aac gct cgt cag aac ctg gcc act ttc tgc        192
Glu Leu Tyr Leu Asp Gly Asn Ala Arg Gln Asn Leu Ala Thr Phe Cys           
    50                  55                  60                            

cag acc tgg gac gac gaa aac gtc cat aaa ttg atg gat ttg tcg atc        240
Gln Thr Trp Asp Asp Glu Asn Val His Lys Leu Met Asp Leu Ser Ile           
65                  70                  75                  80            

aat aaa aac tgg atc gac aaa gaa gaa tat ccg caa tcc gca gcc atc        288
Asn Lys Asn Trp Ile Asp Lys Glu Glu Tyr Pro Gln Ser Ala Ala Ile           
                85                  90                  95                

gac ctg cgt tgc gta aat atg gtt gcc gat ctg tgg cat gcg cct gcg        336
Asp Leu Arg Cys Val Asn Met Val Ala Asp Leu Trp His Ala Pro Ala           
            100                 105                 110                   

ccg aaa aat ggt cag gcc gtt ggc acc aac acc att ggt tct tcc gag        384
Pro Lys Asn Gly Gln Ala Val Gly Thr Asn Thr Ile Gly Ser Ser Glu           
        115                 120                 125                       

gcc tgt atg ctc ggc ggg atg gcg atg aaa tgg cgt tgg cgc aag cgt        432
Ala Cys Met Leu Gly Gly Met Ala Met Lys Trp Arg Trp Arg Lys Arg           
    130                 135                 140                           

atg gaa gct gca ggc aaa cca acg gat aaa cca aac ctg gtg tgc ggt        480
Met Glu Ala Ala Gly Lys Pro Thr Asp Lys Pro Asn Leu Val Cys Gly           
145                 150                 155                 160           

ccg gta caa atc tgc tgg cat aaa ttc gcc cgc tac tgg gat gtg gag        528
Pro Val Gln Ile Cys Trp His Lys Phe Ala Arg Tyr Trp Asp Val Glu           
                165                 170                 175               

ctg cgt gag atc cct atg cgc ccc ggt cag ttg ttt atg gac ccg aaa        576
Leu Arg Glu Ile Pro Met Arg Pro Gly Gln Leu Phe Met Asp Pro Lys           
            180                 185                 190                   

cgc atg att gaa gcc tgt gac gaa aac acc atc ggc gtg gtg ccg act        624
Arg Met Ile Glu Ala Cys Asp Glu Asn Thr Ile Gly Val Val Pro Thr           
        195                 200                 205                       

ttc ggc gtg acc tac acc ggt aac tat gag ttc cca caa ccg ctg cac        672
Phe Gly Val Thr Tyr Thr Gly Asn Tyr Glu Phe Pro Gln Pro Leu His           
    210                 215                 220                           

gat gcg ctg gat aaa ttc cag gcc gac acc ggt atc gac atc gac atg        720
Asp Ala Leu Asp Lys Phe Gln Ala Asp Thr Gly Ile Asp Ile Asp Met           
225                 230                 235                 240           

cac atc gac gct gcc agc ggt ggc ttc ctg gca ccg ttc gtc gcc ccg        768
His Ile Asp Ala Ala Ser Gly Gly Phe Leu Ala Pro Phe Val Ala Pro           
                245                 250                 255               

gat atc gtc tgg gac ttc cgc ctg ccg cgt gtg aaa tcg atc agt gct        816
Asp Ile Val Trp Asp Phe Arg Leu Pro Arg Val Lys Ser Ile Ser Ala           
            260                 265                 270                   

tca ggc cat aaa ttc ggt ctg gct ccg ctg ggc tgc ggc tgg gtt atc        864
Ser Gly His Lys Phe Gly Leu Ala Pro Leu Gly Cys Gly Trp Val Ile           
        275                 280                 285                       

tgg cgt gac gaa gaa gcg ctg ccg cag gaa ctg gtg ttc aac gtt gac        912
Trp Arg Asp Glu Glu Ala Leu Pro Gln Glu Leu Val Phe Asn Val Asp           
    290                 295                 300                           

tac ctg ggt ggt caa att ggt act ttt gcc atc aac ttc tcc cgc ccg        960
Tyr Leu Gly Gly Gln Ile Gly Thr Phe Ala Ile Asn Phe Ser Arg Pro           
305                 310                 315                 320           

gcg ggt cag gta att gca cag tac tat gaa ttc ctg cgc ctc ggt cgt       1008
Ala Gly Gln Val Ile Ala Gln Tyr Tyr Glu Phe Leu Arg Leu Gly Arg           
                325                 330                 335               

gaa ggc tat acc aaa gta cag aac gcc tct tac cag gtt gcc gct tat       1056
Glu Gly Tyr Thr Lys Val Gln Asn Ala Ser Tyr Gln Val Ala Ala Tyr           
            340                 345                 350                   

ctg gcg gat gaa atc gcc aaa ctg ggg ccg tat gag ttc atc tgt acg       1104
Leu Ala Asp Glu Ile Ala Lys Leu Gly Pro Tyr Glu Phe Ile Cys Thr           
        355                 360                 365                       

ggt cgc ccg gac gaa ggc atc ccg gcg gtt tgc ttc aaa ctg aaa gat       1152
Gly Arg Pro Asp Glu Gly Ile Pro Ala Val Cys Phe Lys Leu Lys Asp           
    370                 375                 380                           

ggt gaa gat ccg gga tac acc ctg tac gac ctc tct gaa cgt ctg cgt       1200
Gly Glu Asp Pro Gly Tyr Thr Leu Tyr Asp Leu Ser Glu Arg Leu Arg           
385                 390                 395                 400           

ctg cgc ggc tgg cag gtt ccg gcc ttc act ctc ggc ggt gaa gcc acc       1248
Leu Arg Gly Trp Gln Val Pro Ala Phe Thr Leu Gly Gly Glu Ala Thr           
                405                 410                 415               

gac atc gtg gtg atg cgc att atg tgt cgt cgc ggc ttc gaa atg gac       1296
Asp Ile Val Val Met Arg Ile Met Cys Arg Arg Gly Phe Glu Met Asp           
            420                 425                 430                   

ttt gct gaa ctg ttg ctg gaa gac tac aaa gcc tcc ctg aaa tat ctc       1344
Phe Ala Glu Leu Leu Leu Glu Asp Tyr Lys Ala Ser Leu Lys Tyr Leu           
        435                 440                 445                       

agc gat cac ccg aaa ctg cag ggt att gcc cag cag aac agc ttt aaa       1392
Ser Asp His Pro Lys Leu Gln Gly Ile Ala Gln Gln Asn Ser Phe Lys           
    450                 455                 460                           

cac acc tga                                                           1401
His Thr                                                                   
465                                                                       


<210>  2
<211>  466
<212>  PRT
<213>  Escherichia coli

<400>  2

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>  3
<211>  466
<212>  PRT
<213>  Escherichia coli

<400>  3

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>  4
<211>  511
<212>  PRT
<213>  Escherichia coli

<400>  4

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


Gly Phe Phe Ala Ile Thr Ala Ser Met Val Met Ala Val Tyr Glu Tyr 
            20                  25                  30          


Pro Thr Phe Ala Thr Ser Gly Phe Ser Leu Val Phe Phe Leu Leu Leu 
        35                  40                  45              


Gly Gly Ile Leu Trp Phe Ile Pro Val Gly Leu Cys Ala Ala Glu Met 
    50                  55                  60                  


Ala Thr Val Asp Gly Trp Glu Glu Gly Gly Val Phe Ala Trp Val Ser 
65                  70                  75                  80  


Asn Thr Leu Gly Pro Arg Trp Gly Phe Ala Ala Ile Ser Phe Gly Tyr 
                85                  90                  95      


Leu Gln Ile Ala Ile Gly Phe Ile Pro Met Leu Tyr Phe Val Leu Gly 
            100                 105                 110         


Ala Leu Ser Tyr Ile Leu Lys Trp Pro Ala Leu Asn Glu Asp Pro Ile 
        115                 120                 125             


Thr Lys Thr Ile Ala Ala Leu Ile Ile Leu Trp Ala Leu Ala Leu Thr 
    130                 135                 140                 


Gln Phe Gly Gly Thr Lys Tyr Thr Ala Arg Ile Ala Lys Val Gly Phe 
145                 150                 155                 160 


Phe Ala Gly Ile Leu Leu Pro Ala Phe Ile Leu Ile Ala Leu Ala Ala 
                165                 170                 175     


Ile Tyr Leu His Ser Gly Ala Pro Val Ala Ile Glu Met Asp Ser Lys 
            180                 185                 190         


Thr Phe Phe Pro Asp Phe Ser Lys Val Gly Thr Leu Val Val Phe Val 
        195                 200                 205             


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


Asn Glu Met Ser Asn Pro Gly Arg Asp Tyr Pro Leu Ala Met Leu Leu 
225                 230                 235                 240 


Leu Met Val Ala Ala Ile Cys Leu Ser Ser Val Gly Gly Leu Ser Ile 
                245                 250                 255     


Ala Met Val Ile Pro Gly Asn Glu Ile Asn Leu Ser Ala Gly Val Met 
            260                 265                 270         


Gln Thr Phe Thr Val Leu Met Ser His Val Ala Pro Glu Ile Glu Trp 
        275                 280                 285             


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


Ile Ala Ser Trp Ile Val Gly Pro Ser Arg Gly Met Tyr Val Thr Ala 
305                 310                 315                 320 


Gln Lys Asn Leu Leu Pro Ala Ala Phe Ala Lys Met Asn Lys Asn Gly 
                325                 330                 335     


Val Pro Val Thr Leu Val Ile Ser Gln Leu Val Ile Thr Ser Ile Ala 
            340                 345                 350         


Leu Ile Ile Leu Thr Asn Thr Gly Gly Gly Asn Asn Met Ser Phe Leu 
        355                 360                 365             


Ile Ala Leu Ala Leu Thr Val Val Ile Tyr Leu Cys Ala Tyr Phe Met 
    370                 375                 380                 


Leu Phe Ile Gly Tyr Ile Val Leu Val Leu Lys His Pro Asp Leu Lys 
385                 390                 395                 400 


Arg Thr Phe Asn Ile Pro Gly Gly Lys Gly Val Lys Leu Val Val Ala 
                405                 410                 415     


Ile Val Gly Leu Leu Thr Ser Ile Met Ala Phe Ile Val Ser Phe Leu 
            420                 425                 430         


Pro Pro Asp Asn Ile Gln Gly Asp Ser Thr Asp Met Tyr Val Glu Leu 
        435                 440                 445             


Leu Val Val Ser Phe Leu Val Val Leu Ala Leu Pro Phe Ile Leu Tyr 
    450                 455                 460                 


Ala Val His Asp Arg Lys Gly Lys Ala Asn Thr Gly Val Thr Leu Glu 
465                 470                 475                 480 


Pro Ile Asn Ser Gln Asn Ala Pro Lys Gly His Phe Phe Leu His Pro 
                485                 490                 495     


Arg Ala Arg Ser Pro His Tyr Ile Val Met Asn Asp Lys Lys His 
            500                 505                 510     


<210>  5
<211>  29
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PCR Primer

<400>  5
ggccgctgga gatggaccag aagctgtta                                         29


<210>  6
<211>  27
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  PCR Primer

<400>  6
ccaatgcatt caggtgtgtt taaagct                                           27


