                         SEQUENCE LISTING

<110>  Codexis, Inc.
       Cope, Greg
       Clark, Louis
 
<120>  RECOMBINANT HOST CELLS AND PROCESSES FOR PRODUCING 1,3-BUTADIENE 
       THROUGH A 5-HYDROXYPENT-3-ENOATE INTERMEDIATE

<130>  CX5-112USP1

<160>  4     

<170>  PatentIn version 3.5

<210>  1
<211>  512
<212>  PRT
<213>  Marinobacter algicola strain DG893

<400>  1

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


Glu Gln Leu Arg Gly Lys His Val Leu Ile Thr Gly Thr Thr Gly Phe 
            20                  25                  30          


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


Gly Gly Ile His Leu Leu Ile Arg Gly Asn Lys Arg His Pro Ala Ala 
    50                  55                  60                  


Arg Glu Arg Phe Leu Asn Glu Ile Ala Ser Ser Ser Val Phe Glu Arg 
65                  70                  75                  80  


Leu Arg His Asp Asp Asn Glu Ala Phe Glu Thr Phe Leu Glu Glu Arg 
                85                  90                  95      


Val His Cys Ile Thr Gly Glu Val Thr Glu Ser Arg Phe Gly Leu Thr 
            100                 105                 110         


Pro Glu Arg Phe Arg Ala Leu Ala Gly Gln Val Asp Ala Phe Ile Asn 
        115                 120                 125             


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


Ile Asn Thr Leu Cys Leu Glu Asn Val Ala Ala Leu Ala Glu Leu Asn 
145                 150                 155                 160 


Ser Ala Met Ala Val Ile Gln Val Ser Thr Cys Tyr Val Asn Gly Lys 
                165                 170                 175     


Asn Ser Gly Gln Ile Thr Glu Ser Val Ile Lys Pro Ala Gly Glu Ser 
            180                 185                 190         


Ile Pro Arg Ser Thr Asp Gly Tyr Tyr Glu Ile Glu Glu Leu Val His 
        195                 200                 205             


Leu Leu Gln Asp Lys Ile Ser Asp Val Lys Ala Arg Tyr Ser Gly Lys 
    210                 215                 220                 


Val Leu Glu Lys Lys Leu Val Asp Leu Gly Ile Arg Glu Ala Asn Asn 
225                 230                 235                 240 


Tyr Gly Trp Ser Asp Thr Tyr Thr Phe Thr Lys Trp Leu Gly Glu Gln 
                245                 250                 255     


Leu Leu Met Lys Ala Leu Ser Gly Arg Ser Leu Thr Ile Val Arg Pro 
            260                 265                 270         


Ser Ile Ile Glu Ser Ala Leu Glu Glu Pro Ser Pro Gly Trp Ile Glu 
        275                 280                 285             


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


Val Ser Leu Phe Pro Gly Lys Arg Ser Gly Ile Ile Asp Val Ile Pro 
305                 310                 315                 320 


Val Asp Leu Val Ala Asn Ser Ile Ile Leu Ser Leu Ala Glu Ala Leu 
                325                 330                 335     


Ser Gly Ser Gly Gln Arg Arg Ile Tyr Gln Cys Cys Ser Gly Gly Ser 
            340                 345                 350         


Asn Pro Ile Ser Leu Gly Lys Phe Ile Asp Tyr Leu Met Ala Glu Ala 
        355                 360                 365             


Lys Thr Asn Tyr Ala Ala Tyr Asp Gln Leu Phe Tyr Arg Arg Pro Thr 
    370                 375                 380                 


Lys Pro Phe Val Ala Val Asn Arg Lys Leu Phe Asp Val Val Val Gly 
385                 390                 395                 400 


Gly Met Arg Val Pro Leu Ser Ile Ala Gly Lys Ala Met Arg Leu Ala 
                405                 410                 415     


Gly Gln Asn Arg Glu Leu Lys Val Leu Lys Asn Leu Asp Thr Thr Arg 
            420                 425                 430         


Ser Leu Ala Thr Ile Phe Gly Phe Tyr Thr Ala Pro Asp Tyr Ile Phe 
        435                 440                 445             


Arg Asn Asp Ser Leu Met Ala Leu Ala Ser Arg Met Gly Glu Leu Asp 
    450                 455                 460                 


Arg Val Leu Phe Pro Val Asp Ala Arg Gln Ile Asp Trp Gln Leu Tyr 
465                 470                 475                 480 


Leu Cys Lys Ile His Leu Gly Gly Leu Asn Arg Tyr Ala Leu Lys Glu 
                485                 490                 495     


Arg Lys Leu Tyr Ser Leu Arg Ala Ala Asp Thr Arg Lys Lys Ala Ala 
            500                 505                 510         


<210>  2
<211>  513
<212>  PRT
<213>  Marinobacter aquaeolei

<400>  2

Met Ala Ile Gln Gln Val His His Ala Asp Thr Ser Ser Ser Lys Val 
1               5                   10                  15      


Leu Gly Gln Leu Arg Gly Lys Arg Val Leu Ile Thr Gly Thr Thr Gly 
            20                  25                  30          


Phe Leu Gly Lys Val Val Leu Glu Arg Leu Ile Arg Ala Val Pro Asp 
        35                  40                  45              


Ile Gly Ala Ile Tyr Leu Leu Ile Arg Gly Asn Lys Arg His Pro Asp 
    50                  55                  60                  


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


Arg Leu Arg Glu Ala Asp Ser Glu Gly Phe Asp Ala Phe Leu Glu Glu 
                85                  90                  95      


Arg Ile His Cys Val Thr Gly Glu Val Thr Glu Ala Gly Phe Gly Ile 
            100                 105                 110         


Gly Gln Glu Asp Tyr Arg Lys Leu Ala Thr Glu Leu Asp Ala Val Ile 
        115                 120                 125             


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


Ala Ile Asn Thr Leu Cys Leu Arg Asn Ile Ala Gly Met Val Asp Leu 
145                 150                 155                 160 


Asn Pro Lys Leu Ala Val Leu Gln Val Ser Thr Cys Tyr Val Asn Gly 
                165                 170                 175     


Met Asn Ser Gly Gln Val Thr Glu Ser Val Ile Lys Pro Ala Gly Glu 
            180                 185                 190         


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


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


Lys Val Leu Glu Arg Lys Leu Val Asp Leu Gly Ile Arg Glu Ala Asn 
225                 230                 235                 240 


Arg Tyr Gly Trp Ser Asp Thr Tyr Thr Phe Thr Lys Trp Leu Gly Glu 
                245                 250                 255     


Gln Leu Leu Met Lys Ala Leu Asn Gly Arg Thr Leu Thr Ile Leu Arg 
            260                 265                 270         


Pro Ser Ile Ile Glu Ser Ala Leu Glu Glu Pro Ala Pro Gly Trp Ile 
        275                 280                 285             


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


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


Pro Val Asp Leu Val Ala Asn Ser Ile Ile Leu Ser Leu Ala Glu Ala 
                325                 330                 335     


Leu Gly Glu Pro Gly Arg Arg Arg Ile Tyr Gln Cys Cys Ser Gly Gly 
            340                 345                 350         


Gly Asn Pro Ile Ser Leu Gly Glu Phe Ile Asp His Leu Met Ala Glu 
        355                 360                 365             


Ser Lys Ala Asn Tyr Ala Ala Tyr Asp His Leu Phe Tyr Arg Gln Pro 
    370                 375                 380                 


Ser Lys Pro Phe Leu Ala Val Asn Arg Ala Leu Phe Asp Leu Val Ile 
385                 390                 395                 400 


Ser Gly Val Arg Leu Pro Leu Ser Leu Thr Asp Arg Val Leu Lys Leu 
                405                 410                 415     


Leu Gly Asn Ser Arg Asp Leu Lys Met Leu Arg Asn Leu Asp Thr Thr 
            420                 425                 430         


Gln Ser Leu Ala Thr Ile Phe Gly Phe Tyr Thr Ala Pro Asp Tyr Ile 
        435                 440                 445             


Phe Arg Asn Asp Glu Leu Met Ala Leu Ala Asn Arg Met Gly Glu Val 
    450                 455                 460                 


Asp Lys Gly Leu Phe Pro Val Asp Ala Arg Leu Ile Asp Trp Glu Leu 
465                 470                 475                 480 


Tyr Leu Arg Lys Ile His Leu Ala Gly Leu Asn Arg Tyr Ala Leu Lys 
                485                 490                 495     


Glu Arg Lys Val Tyr Ser Leu Lys Thr Ala Arg Gln Arg Lys Lys Ala 
            500                 505                 510         


Ala 
    


<210>  3
<211>  514
<212>  PRT
<213>  Oceanobacter sp. strain RED65

<400>  3

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


His Ile Phe Leu Thr Gly Val Thr Gly Phe Leu Gly Lys Ala Ile Leu 
            20                  25                  30          


Glu Lys Leu Leu Tyr Ser Val Pro Gln Leu Ala Gln Ile His Ile Leu 
        35                  40                  45              


Val Arg Gly Gly Lys Val Ser Ala Lys Lys Arg Phe Gln His Asp Ile 
    50                  55                  60                  


Leu Gly Ser Ser Ile Phe Glu Arg Leu Lys Glu Gln His Gly Glu His 
65                  70                  75                  80  


Phe Glu Glu Trp Val Gln Ser Lys Ile Asn Leu Val Glu Gly Glu Leu 
                85                  90                  95      


Thr Gln Pro Met Phe Asp Leu Pro Ser Ala Glu Phe Ala Gly Leu Ala 
            100                 105                 110         


Asn Gln Leu Asp Leu Ile Ile Asn Ser Ala Ala Ser Val Asn Phe Arg 
        115                 120                 125             


Glu Asn Leu Glu Lys Ala Leu Asn Ile Asn Thr Leu Cys Leu Asn Asn 
    130                 135                 140                 


Ile Ile Ala Leu Ala Gln Tyr Asn Val Ala Ala Gln Thr Pro Val Met 
145                 150                 155                 160 


Gln Ile Ser Thr Cys Tyr Val Asn Gly Phe Asn Lys Gly Gln Ile Asn 
                165                 170                 175     


Glu Glu Val Val Gly Pro Ala Ser Gly Leu Ile Pro Gln Leu Ser Gln 
            180                 185                 190         


Asp Cys Tyr Asp Ile Asp Ser Val Phe Lys Arg Val His Ser Gln Ile 
        195                 200                 205             


Glu Gln Val Lys Lys Arg Lys Thr Asp Ile Glu Gln Gln Glu Gln Ala 
    210                 215                 220                 


Leu Ile Lys Leu Gly Ile Lys Thr Ser Gln His Phe Gly Trp Asn Asp 
225                 230                 235                 240 


Thr Tyr Thr Phe Thr Lys Trp Leu Gly Glu Gln Leu Leu Ile Gln Lys 
                245                 250                 255     


Leu Gly Lys Gln Ser Leu Thr Ile Leu Arg Pro Ser Ile Ile Glu Ser 
            260                 265                 270         


Ala Val Arg Glu Pro Ala Pro Gly Trp Val Glu Gly Val Lys Val Ala 
        275                 280                 285             


Asp Ala Leu Ile Tyr Ala Tyr Ala Lys Gly Arg Val Ser Ile Phe Pro 
    290                 295                 300                 


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


Asn Ala Ala Ala Leu Ser Ala Ala Gln Leu Met Glu Ser Asn Gln Gln 
                325                 330                 335     


Thr Gly Tyr Arg Ile Tyr Gln Cys Cys Ser Gly Ser Arg Asn Pro Ile 
            340                 345                 350         


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


Tyr Gln Glu Trp Pro Lys Leu Phe Ala Asp Lys Pro Gln Glu Ala Phe 
    370                 375                 380                 


Lys Thr Val Ser Pro Lys Arg Phe Lys Leu Tyr Met Ser Gly Phe Thr 
385                 390                 395                 400 


Ala Ile Thr Trp Ala Lys Thr Ile Ile Gly Arg Val Phe Gly Ser Asn 
                405                 410                 415     


Ala Ala Ser Gln His Met Leu Lys Ala Lys Thr Thr Ala Ser Leu Ala 
            420                 425                 430         


Asn Ile Phe Gly Phe Tyr Thr Ala Pro Asn Tyr Arg Phe Ser Ser Gln 
        435                 440                 445             


Lys Leu Glu Gln Leu Val Lys Gln Phe Asp Thr Thr Glu Gln Arg Leu 
    450                 455                 460                 


Tyr Asp Ile Arg Ala Asp His Phe Asp Trp Lys Tyr Tyr Leu Gln Glu 
465                 470                 475                 480 


Val His Met Asp Gly Leu His Lys Tyr Ala Leu Ala Asp Arg Gln Glu 
                485                 490                 495     


Leu Lys Pro Lys His Val Lys Lys Arg Lys Arg Glu Thr Ile Arg Gln 
            500                 505                 510         


Ala Ala 
        


<210>  4
<211>  460
<212>  PRT
<213>  Bombyx mori

<400>  4

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


Phe Tyr Asp Gly Lys Ser Val Phe Ile Thr Gly Ala Thr Gly Phe Leu 
            20                  25                  30          


Gly Lys Ala Tyr Val Glu Lys Leu Ala Tyr Ser Cys Pro Gly Ile Val 
        35                  40                  45              


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


Arg Met Arg Lys Tyr Leu Asp Gln Pro Ile Phe Ser Arg Ile Lys Tyr 
65                  70                  75                  80  


Glu His Pro Glu Tyr Phe Lys Lys Ile Ile Pro Ile Ser Gly Asp Ile 
                85                  90                  95      


Thr Ala Pro Lys Leu Gly Leu Cys Asp Glu Glu Arg Asn Ile Leu Ile 
            100                 105                 110         


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


Asp His Leu Lys Phe Thr Leu Asn Thr Asn Val Gly Gly Thr Met Lys 
    130                 135                 140                 


Val Leu Glu Leu Val Lys Glu Met Lys Asn Leu Ala Met Phe Val Tyr 
145                 150                 155                 160 


Val Ser Thr Ala Tyr Ser Asn Thr Ser Gln Arg Ile Leu Glu Glu Lys 
                165                 170                 175     


Leu Tyr Pro Gln Ser Leu Asn Leu Asn Glu Ile Gln Lys Phe Ala Glu 
            180                 185                 190         


Glu His Tyr Ile Leu Gly Lys Asp Asn Asp Glu Met Ile Lys Phe Ile 
        195                 200                 205             


Gly Asn His Pro Asn Thr Tyr Ala Tyr Thr Lys Ala Leu Ala Glu Asn 
    210                 215                 220                 


Leu Val Ala Glu Glu His Gly Glu Ile Pro Thr Ile Ile Ile Arg Pro 
225                 230                 235                 240 


Ser Ile Ile Thr Ala Ser Ala Glu Glu Pro Val Arg Gly Phe Val Asp 
                245                 250                 255     


Ser Trp Ser Gly Ala Thr Ala Met Ala Ala Phe Ala Leu Lys Gly Trp 
            260                 265                 270         


Asn Asn Ile Met Tyr Ser Thr Gly Glu Glu Asn Ile Asp Leu Ile Pro 
        275                 280                 285             


Leu Asp Tyr Val Val Asn Leu Thr Leu Val Ala Ile Ala Lys Tyr Lys 
    290                 295                 300                 


Pro Thr Lys Glu Val Thr Val Tyr His Val Thr Thr Ser Asp Leu Asn 
305                 310                 315                 320 


Pro Ile Ser Ile Arg Arg Ile Phe Ile Lys Leu Ser Glu Phe Ala Ser 
                325                 330                 335     


Lys Asn Pro Thr Ser Asn Ala Ala Pro Phe Ala Ala Thr Thr Leu Leu 
            340                 345                 350         


Thr Lys Gln Lys Pro Leu Ile Lys Leu Val Thr Phe Leu Met Gln Thr 
        355                 360                 365             


Thr Pro Ala Phe Leu Ala Asp Leu Trp Met Lys Thr Gln Arg Lys Glu 
    370                 375                 380                 


Ala Lys Phe Val Lys Gln His Asn Leu Val Val Arg Ser Arg Asp Gln 
385                 390                 395                 400 


Leu Glu Phe Phe Thr Ser Gln Ser Trp Leu Leu Arg Cys Glu Arg Ala 
                405                 410                 415     


Arg Val Leu Ser Ala Ala Leu Ser Asp Ser Asp Arg Ala Val Phe Arg 
            420                 425                 430         


Cys Asp Pro Ser Thr Ile Asp Trp Asp Gln Tyr Leu Pro Ile Tyr Phe 
        435                 440                 445             


Glu Gly Ile Asn Lys His Leu Phe Lys Asn Lys Leu 
    450                 455                 460 


