                         SEQUENCE LISTING

<110>  Massachusetts Institute of Technology
       Budde, Charles
       Rha, ChoKyun
       Sinskey, Anthony
 
<120>  METHODS FOR PRODUCING POLYHYDROXYALKANOATE COPOLYMER WITH HIGH 
       MEDIUM CHAIN LENGTH MONOMER CONTENT

<130>  M0656.70198WO00

<150>  US 61/260,164
<151>  2009-11-11

<160>  4     

<170>  PatentIn version 3.5

<210>  1
<211>  1689
<212>  DNA
<213>  Rhodococcus aetherivorans I24

<400>  1
ttgctcgacc acgtgcacaa gaagttgaag tcgaccctgg acccgatcgg ctggggtccc       60

gcggtgaagt cggtggccgg acgcgccgtc cgcaaccccc aggccgtcac cgccgccacg      120

acggaatacg cgggccggct ggtgaagatc cccgcggcgg ccacccgcgt gttcaacgcc      180

gacgatccca agccgccgat gccgctcgac ccgcgggacc gccgtttctc cgacaccgcc      240

tggcgggaga accccgcgta cttctcgctc ctgcagagtt atctcgcgac gcgggcctac      300

gtcgaggaac tcaccgacgc cggcgccggc gatccgctgc aggacggcaa ggcccgccag      360

ttcgcgaacc tgatgctcga cgtgctggcc ccgtcgaact tcctgtggaa tccgggcgtg      420

ctcacccgtg cattcgagac gggcggggca agcctgctgc gcggcgcccg atatgccgtg      480

cacgacgtgc tcaaccgcgg cggcctgccg ctgaaggtgg actcggacgc gttcaccgtc      540

ggcgagaacc tcgcggccac cccgggcaag gtggtctatc gcaacgacct gatcgagctg      600

atccagtaca cgccgcagac cgagcaggtg catgcggtgc cgatcctcgc cgcgccgccg      660

tggatcaaca agtactacat cctcgatctc gcacccggtc gcagcctcgc cgagtgggcg      720

gtccagcacg gccgcaccgt gttcatgctc tcgtaccgga acccggacga gtcgatgcgg      780

cacatcacca tggacgacta ctacgtcaac ggcattgccg ccgcgctgga cgtggtcgag      840

gagatcaccg ggtcgccgaa gatcgaggtg ctgtccatct gcctcggcgg cgcgatggcc      900

gcgatggccg ccgcgcgcgc attcgccgtc ggcgacaagc gcgtgaccgc cttcaccatg      960

ctcaacaccc tgctcgacta cagccaggtc ggggaactcg ggttgctgac cgatccgtcc     1020

acgctggacc tcgtcgagtt ccggatgcgg cagcagggct tcctgtccgg caaggagatg     1080

gccggcagct tcgacatgat ccgcgcgaag gacctcgtct tcaactactg ggtctcgcgg     1140

tggatgaagg gcgagaagcc tgcggccttc gacatcctcg cgtggaacga ggacagcacg     1200

agcatgcccg cggagatgca ctcgcactac ctccggtcgc tgtacggccg caacgagctg     1260

gccgagggcc tctacgtgct cgacggacag cccctgaacc tgcacgacat cacgtgcgac     1320

acctacgtcg tcggcgcgat caacgaccac atcgtgccct ggacatcgtc gtaccaggcg     1380

gtgaacctgc tgggcggcga cgtgcgctac gtgctcacca acggcgggca cgtcgccggc     1440

gcggtgaacc cgcccggcaa gaaggtgtgg ttcaaggccg tcggggcgcc ggacgccgag     1500

accggctcgc cgctgcccgc ggatccgcag gtctgggacg acgcggccac ccgctacgag     1560

cactcgtggt gggaggactg gacggcctgg tcgaacaagc gcgccgggga gctggtgccg     1620

ccgccggcaa tgggcagcgc cgcccacccg ccgctcgagg acgctccggg cacgtacgtc     1680

ttcagctga                                                             1689


<210>  2
<211>  562
<212>  PRT
<213>  Rhodococcus aetherivorans I24

<400>  2

Met Leu Asp His Val His Lys Lys Leu Lys Ser Thr Leu Asp Pro Ile 
1               5                   10                  15      


Gly Trp Gly Pro Ala Val Lys Ser Val Ala Gly Arg Ala Val Arg Asn 
            20                  25                  30          


Pro Gln Ala Val Thr Ala Ala Thr Thr Glu Tyr Ala Gly Arg Leu Val 
        35                  40                  45              


Lys Ile Pro Ala Ala Ala Thr Arg Val Phe Asn Ala Asp Asp Pro Lys 
    50                  55                  60                  


Pro Pro Met Pro Leu Asp Pro Arg Asp Arg Arg Phe Ser Asp Thr Ala 
65                  70                  75                  80  


Trp Arg Glu Asn Pro Ala Tyr Phe Ser Leu Leu Gln Ser Tyr Leu Ala 
                85                  90                  95      


Thr Arg Ala Tyr Val Glu Glu Leu Thr Asp Ala Gly Ala Gly Asp Pro 
            100                 105                 110         


Leu Gln Asp Gly Lys Ala Arg Gln Phe Ala Asn Leu Met Leu Asp Val 
        115                 120                 125             


Leu Ala Pro Ser Asn Phe Leu Trp Asn Pro Gly Val Leu Thr Arg Ala 
    130                 135                 140                 


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


His Asp Val Leu Asn Arg Gly Gly Leu Pro Leu Lys Val Asp Ser Asp 
                165                 170                 175     


Ala Phe Thr Val Gly Glu Asn Leu Ala Ala Thr Pro Gly Lys Val Val 
            180                 185                 190         


Tyr Arg Asn Asp Leu Ile Glu Leu Ile Gln Tyr Thr Pro Gln Thr Glu 
        195                 200                 205             


Gln Val His Ala Val Pro Ile Leu Ala Ala Pro Pro Trp Ile Asn Lys 
    210                 215                 220                 


Tyr Tyr Ile Leu Asp Leu Ala Pro Gly Arg Ser Leu Ala Glu Trp Ala 
225                 230                 235                 240 


Val Gln His Gly Arg Thr Val Phe Met Leu Ser Tyr Arg Asn Pro Asp 
                245                 250                 255     


Glu Ser Met Arg His Ile Thr Met Asp Asp Tyr Tyr Val Asn Gly Ile 
            260                 265                 270         


Ala Ala Ala Leu Asp Val Val Glu Glu Ile Thr Gly Ser Pro Lys Ile 
        275                 280                 285             


Glu Val Leu Ser Ile Cys Leu Gly Gly Ala Met Ala Ala Met Ala Ala 
    290                 295                 300                 


Ala Arg Ala Phe Ala Val Gly Asp Lys Arg Val Thr Ala Phe Thr Met 
305                 310                 315                 320 


Leu Asn Thr Leu Leu Asp Tyr Ser Gln Val Gly Glu Leu Gly Leu Leu 
                325                 330                 335     


Thr Asp Pro Ser Thr Leu Asp Leu Val Glu Phe Arg Met Arg Gln Gln 
            340                 345                 350         


Gly Phe Leu Ser Gly Lys Glu Met Ala Gly Ser Phe Asp Met Ile Arg 
        355                 360                 365             


Ala Lys Asp Leu Val Phe Asn Tyr Trp Val Ser Arg Trp Met Lys Gly 
    370                 375                 380                 


Glu Lys Pro Ala Ala Phe Asp Ile Leu Ala Trp Asn Glu Asp Ser Thr 
385                 390                 395                 400 


Ser Met Pro Ala Glu Met His Ser His Tyr Leu Arg Ser Leu Tyr Gly 
                405                 410                 415     


Arg Asn Glu Leu Ala Glu Gly Leu Tyr Val Leu Asp Gly Gln Pro Leu 
            420                 425                 430         


Asn Leu His Asp Ile Thr Cys Asp Thr Tyr Val Val Gly Ala Ile Asn 
        435                 440                 445             


Asp His Ile Val Pro Trp Thr Ser Ser Tyr Gln Ala Val Asn Leu Leu 
    450                 455                 460                 


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


Ala Val Asn Pro Pro Gly Lys Lys Val Trp Phe Lys Ala Val Gly Ala 
                485                 490                 495     


Pro Asp Ala Glu Thr Gly Ser Pro Leu Pro Ala Asp Pro Gln Val Trp 
            500                 505                 510         


Asp Asp Ala Ala Thr Arg Tyr Glu His Ser Trp Trp Glu Asp Trp Thr 
        515                 520                 525             


Ala Trp Ser Asn Lys Arg Ala Gly Glu Leu Val Pro Pro Pro Ala Met 
    530                 535                 540                 


Gly Ser Ala Ala His Pro Pro Leu Glu Asp Ala Pro Gly Thr Tyr Val 
545                 550                 555                 560 


Phe Ser 
        


<210>  3
<211>  1686
<212>  DNA
<213>  Rhodococcus aetherivorans I24

<400>  3
atgatggccc aggcacgaac cgtgatcggt gagagcgtcg aggagtcgat cgggggtggc       60

gaggacgtcg cgccaccgag gctcgggccg gccgtcggcg ccctggccga cgtgttcggt      120

cacggccggg cggtggcccg gcacggcgtg tcgttcggca gggaactggc gaagatcgcc      180

gtcggccggt cgacggtggc tccggcgaag ggagaccgcc ggttcgccga ctcggcgtgg      240

agtgcgaacc ccgcctaccg ccggctcggg cagacctacc tggcggcaac cgaggccgtc      300

gacggagtcg tcgacgaggt cggtcgcgcg atcggcccgc gacgcacggc cgaggccagg      360

ttcgccgccg acatcctcac cgcggccctg gccccgacga actacctgtg gaccaacccc      420

gcggcgctga aggaggcgtt cgacaccgcc ggactcagcc tcgcacgcgg caccaagcac      480

ttcgtctccg atctgatcga gaaccggggc atgccgtcga tggtccagcg cggcgccttc      540

accgtcggga aggaccttgc ggtgaccccg ggtgcggtga tctcccgcga cgaggtcgcc      600

gaggtgctgc agtacacccc gaccacggag acggtccgcc gccggccggt gctcgtggtg      660

cccccgccga tcggccggta ctacttcctg gacctgcggc cgggacgcag cttcgtcgag      720

tacagcgtgg gccggggcct gcagaccttc ctgctgtcgt ggcgcaatcc caccgccgag      780

cagggcgact gggacttcga cacgtacgcg ggccgggtga tccgggcgat cgacgaggtg      840

cgggagatca ccggcagcga cgacgtgaac ctgatcggtt tctgcgccgg cgggatcatc      900

gccaccacgg tgctcaatca ccttgccgcg cagggcgaca cccgagtgca cagcatggcc      960

tatgcggtga cgatgctgga cttcggcgat ccggcactgc tcggcgcgtt cgcccggccc     1020

ggcctgatcc ggttcgccaa gggccggtcc cgccgcaagg gcatcatcag cgcccgcgac     1080

atggggtccg cgttcacctg gatgcgcccg aacgacctgg tgttcaacta cgtcgtcaac     1140

aactacctca tgggtcgcac cccaccggcc ttcgacatcc tcgcctggaa cgacgacggc     1200

accaacctgc ccggcgccct gcacggtcag ttcctcgaca tcttccgtga caacgtgctc     1260

gtcgagcccg gccggctcgc cgtgctgggc acgcccgtcg acctgaagtc gatcaccgtg     1320

cccacgttcg tctcgggcgc catcgccgac catctgaccg catggcgcaa ctgctaccgc     1380

accacccaat tgctcggtgg agaaacagaa ttcgcgctca gcttctccgg gcacatcgcc     1440

agcctggtca acccgccggg caatccgaag gcacactact ggaccggggg cacacccggc     1500

ccggacccgg atgcctggct cgagaacgcc gagcggcagc agggcagctg gtggcaggcc     1560

tggtccgact gggtgctcgc ccgcggcggg gaggaaaccg ccgcgccgga cgcacccggc     1620

agtgcgcagc atcccgcgct cgacgccgct cccggccggt acgtgcgcga cctgcccgcc     1680

ggctga                                                                1686


<210>  4
<211>  561
<212>  PRT
<213>  Rhodococcus aetherivorans I24

<400>  4

Met Met Ala Gln Ala Arg Thr Val Ile Gly Glu Ser Val Glu Glu Ser 
1               5                   10                  15      


Ile Gly Gly Gly Glu Asp Val Ala Pro Pro Arg Leu Gly Pro Ala Val 
            20                  25                  30          


Gly Ala Leu Ala Asp Val Phe Gly His Gly Arg Ala Val Ala Arg His 
        35                  40                  45              


Gly Val Ser Phe Gly Arg Glu Leu Ala Lys Ile Ala Val Gly Arg Ser 
    50                  55                  60                  


Thr Val Ala Pro Ala Lys Gly Asp Arg Arg Phe Ala Asp Ser Ala Trp 
65                  70                  75                  80  


Ser Ala Asn Pro Ala Tyr Arg Arg Leu Gly Gln Thr Tyr Leu Ala Ala 
                85                  90                  95      


Thr Glu Ala Val Asp Gly Val Val Asp Glu Val Gly Arg Ala Ile Gly 
            100                 105                 110         


Pro Arg Arg Thr Ala Glu Ala Arg Phe Ala Ala Asp Ile Leu Thr Ala 
        115                 120                 125             


Ala Leu Ala Pro Thr Asn Tyr Leu Trp Thr Asn Pro Ala Ala Leu Lys 
    130                 135                 140                 


Glu Ala Phe Asp Thr Ala Gly Leu Ser Leu Ala Arg Gly Thr Lys His 
145                 150                 155                 160 


Phe Val Ser Asp Leu Ile Glu Asn Arg Gly Met Pro Ser Met Val Gln 
                165                 170                 175     


Arg Gly Ala Phe Thr Val Gly Lys Asp Leu Ala Val Thr Pro Gly Ala 
            180                 185                 190         


Val Ile Ser Arg Asp Glu Val Ala Glu Val Leu Gln Tyr Thr Pro Thr 
        195                 200                 205             


Thr Glu Thr Val Arg Arg Arg Pro Val Leu Val Val Pro Pro Pro Ile 
    210                 215                 220                 


Gly Arg Tyr Tyr Phe Leu Asp Leu Arg Pro Gly Arg Ser Phe Val Glu 
225                 230                 235                 240 


Tyr Ser Val Gly Arg Gly Leu Gln Thr Phe Leu Leu Ser Trp Arg Asn 
                245                 250                 255     


Pro Thr Ala Glu Gln Gly Asp Trp Asp Phe Asp Thr Tyr Ala Gly Arg 
            260                 265                 270         


Val Ile Arg Ala Ile Asp Glu Val Arg Glu Ile Thr Gly Ser Asp Asp 
        275                 280                 285             


Val Asn Leu Ile Gly Phe Cys Ala Gly Gly Ile Ile Ala Thr Thr Val 
    290                 295                 300                 


Leu Asn His Leu Ala Ala Gln Gly Asp Thr Arg Val His Ser Met Ala 
305                 310                 315                 320 


Tyr Ala Val Thr Met Leu Asp Phe Gly Asp Pro Ala Leu Leu Gly Ala 
                325                 330                 335     


Phe Ala Arg Pro Gly Leu Ile Arg Phe Ala Lys Gly Arg Ser Arg Arg 
            340                 345                 350         


Lys Gly Ile Ile Ser Ala Arg Asp Met Gly Ser Ala Phe Thr Trp Met 
        355                 360                 365             


Arg Pro Asn Asp Leu Val Phe Asn Tyr Val Val Asn Asn Tyr Leu Met 
    370                 375                 380                 


Gly Arg Thr Pro Pro Ala Phe Asp Ile Leu Ala Trp Asn Asp Asp Gly 
385                 390                 395                 400 


Thr Asn Leu Pro Gly Ala Leu His Gly Gln Phe Leu Asp Ile Phe Arg 
                405                 410                 415     


Asp Asn Val Leu Val Glu Pro Gly Arg Leu Ala Val Leu Gly Thr Pro 
            420                 425                 430         


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


Ala Asp His Leu Thr Ala Trp Arg Asn Cys Tyr Arg Thr Thr Gln Leu 
    450                 455                 460                 


Leu Gly Gly Glu Thr Glu Phe Ala Leu Ser Phe Ser Gly His Ile Ala 
465                 470                 475                 480 


Ser Leu Val Asn Pro Pro Gly Asn Pro Lys Ala His Tyr Trp Thr Gly 
                485                 490                 495     


Gly Thr Pro Gly Pro Asp Pro Asp Ala Trp Leu Glu Asn Ala Glu Arg 
            500                 505                 510         


Gln Gln Gly Ser Trp Trp Gln Ala Trp Ser Asp Trp Val Leu Ala Arg 
        515                 520                 525             


Gly Gly Glu Glu Thr Ala Ala Pro Asp Ala Pro Gly Ser Ala Gln His 
    530                 535                 540                 


Pro Ala Leu Asp Ala Ala Pro Gly Arg Tyr Val Arg Asp Leu Pro Ala 
545                 550                 555                 560 


Gly 
    


