               SEQUENCE LISTING

<110> CONSEJO SUPERIOR DE INVESTIGACIONES CIENTIFICAS

<120> SISTEMA PARA MEJORAR LA PRODUCCIÓN DE POLIHIDROXIALCANOATOS (BIOPLÁSTICO) POR FERMENTACIÓN A PARTIR DE GLICEROL UTILIZANDO UNA CEPA DE Pseudomonas putida MODIFICADA GENÉTICAMENTE

<130> 5110259 AUXI PRIETO

<160> 8

<170> BiSSAP 1.0

<210> 1
<211> 5021
<212> DNA
<213> Pseudomonas putida KT2440

<220> 
<221> source
<222> 1..5021
<223> /mol_type="DNA"
      /note="Secuencia nucleotidica del cluster glp (Secuencia reversa 
      complementaria desde la posicion 1231166 a la 1236186)"
      /organism="Pseudomonas putida KT2440"

<220> 
<221> gene glpR
<222> 2505..3260

<400> 1
atgacgactg ctctgcgcca acccacactg tccagccaat gcctggccga gtttctcggc       60

accgccctgc tcatcttctt cggtaccggc tgcgtcgccg cgctcaaggt ggccggcgcc      120

agcttcgggc tttgggaaat cagtatcatc tggggcgtgg gcgtcagcat ggccatctac      180

ctcaccgccg gtatttccgg tgcgcacctg aacccagccg tgagtatcgc cctcacactg      240

ttcgcagggt tcgacaagcg caagctgccc ttctacatgc tggcccaggt atgcggcgca      300

ttctgtggcg cagcactggt ctacacgctg tacagcaatc tgttcttcga tttcgaacaa      360

gcccacgcca tgctgcgcgg tagcgaaggc agcctggagc tggcctcggt gttctccacc      420

tacccgcacc cgtcgctgtc caccagccag gcgtttctgg ttgaagtggt gatcactgcc      480

attctgatgg ccgtgatcat ggccctgacc gacgacaaca acggcctgcc gcgcggcgcc      540

atggccccgc tgctgatcgg cctgctgatt gcagtgattg gcagcgccat gggcccattg      600

actggctttg cgatgaaccc ggcccgcgat ttcgggccaa aactcatgac cttcctggcc      660

ggttggggcg aaatcgcctt cactggcggg cgggatattc cctatttcct ggttccggta      720

tttgcaccga tccttggcgc ctgcttgggt gcagcgagct atcgcggcct gattgcacgc      780

aacctgccaa tggcgcccgc cgcgacccct gagacaaatg acattcgcca gggcgatact      840

caagccaatt gatgccacca tcacgcacag ccctgccccc ctattcttgc aaggcctacg      900

accatgacag acacccagga taagaactac atcatcgccc tggaccaggg taccaccagt      960

tcgcgggcga tcattttcga ccgtgacgcc aatgttgtag gtacctccca gcgcgagttc     1020

gcacagcatt acccgcaggc gggctgggtc gagcatgacc cgatggagat tttcgccaca     1080

caaagcgcca ccatggtcga ggcccttgcc caggcgggta tcagccacgc ccaggtcgct     1140

gcgctgggca tcaccaacca acgcgaaacc acggtggtgt gggacaagga aaccgggcgt     1200

ccggtgtaca acgccattgt ctggcagtgc cgccgcagca ccgagatctg tgctcagctc     1260

aagcgtgacg gccatgaaga ctacattcgt gaaaccaccg gcctggtcac cgacccgtat     1320

ttctccggca ccaagctgaa atggatcctc gacaatgtcg acggcgcccg tgaacgcgcc     1380

gagcgtggcg aactgctgtt cggcaccatc gacacctggc tgatctggaa gcttacgggt     1440

gggaaggttc acgtcaccga ctacaccaat gcctcgcgca cgctgatgtt caacatccac     1500

agcctgcagt gggacgacaa gctgctggaa atcctgggca tcccgcggca gatgctgcct     1560

gaggtgcgcc catcctcgga agtgtacggc cacaccaaga gcggcatagc catcgccggt     1620

atcgccggcg accagcaatc ggcactgttc ggccagatgt gcgtggagcc gggccaagcc     1680

aagaatacct acggcactgg ctgcttcctg ttgatgaaca cgggcgacca ggcggtgaag     1740

tcgtcccacg gcctgctcac gaccattgcc tgcggcccgc gtggtgaggt ggcctatgca     1800

ctggaaggcg cggtattcaa tggtggctct accgtgcaat ggctgcgtga cgaactgaag     1860

atcgtcaacg acgccctgga taccgagtat ttcgccagca aggtcaagga cagcaatggc     1920

gtgtacctgg tgccagcctt taccggcctg ggtgccccgt actgggaccc gtatgcccgt     1980

ggtgcgttgt tcggcctgac acgtggcgtg aaggtagacc acatcatccg cgccgcactg     2040

gagtccatcg cctaccagac ccgcgatgtc ctcgacgcca tgcagcagga ctgtggccag     2100

cgcctgtccg agctgcgcgt ggacggtggt gccgtggcca acaacttcct catgcagttc     2160

caggccgaca tcctcggcac ctgcgtggag cggccgcaga tgcgcgaaac cacggcgctg     2220

ggcgccgcct atctggcagg cctggcctgt ggcttctgga gtggcctgga cgagctgcgc     2280

gacaaggcga tcatcgagcg tgagttcagc ccgcaactgg acgaaacgca gaaagaaaag     2340

ctgtatgccg gctggaagaa ggcggtcgag cgtacccgtg actgggaaga tcacgaggca     2400

tgatagctgt tggggccgcc atgcggcccc aatcgtctac tggccgtcat ccccttccta     2460

cggcatcatt gcagaatttg tccggcagcc ccaaaggacc gcccatgaat ctgccccccc     2520

gccaacaaca aatcctcgaa ctggtacgcg aacgcggcta tgtcagtatc gaggaaatgg     2580

cgcagctgtt cgttgtcact ccccagacca tccgacgcga catcaatcag cttgccgagc     2640

tcaacctgct gcgccgttac cacggcggcg cagcttacga ctcgagcatc gagaacaccg     2700

cctacgccat gcgtgccgac cagatgcgtg acgaaaagca gcgcattgcc gacgccgtag     2760

cccggcagat tcccgaccat gcgtcgctgt tcatcaacat cggcaccacc accgaatcga     2820

tcgcacgggc gctgctcaac cacagccacc tgaaagtcat caccaataac ttgcatgtgg     2880

cggcgatcct ggccgccaag gacgatttcg aagtgctggt ggcaggtggc acggtgcgcc     2940

gagatggtgg tgtcgtcggc caggccagtg tcgatttcat caaccagttc aaggtcgact     3000

tcgccctggt gggcatcagc ggtatcgacg aggatggcag cctgctcgat ttcgactatc     3060

aggaagtgcg ggtgtcgcaa gccatcattg ccaatgcccg gcaagtgatc ctcgccgccg     3120

actccagcaa gttcgggcgc aatgccatgg tgcgcctggg ctcgatcgac ctggtcgact     3180

gcctggtgac cgaccggaca ccaaccccgg cccttaccca attgttgaac cagaacaaga     3240

tcaggcttga ggtggtctga gggctggtgg gtgcatgccg ccagactttt ggtgtctgcg     3300

tgagcaagcg ccccctcacc caagccgtac cccctgaatt ttcgtaaatg ttcacttcat     3360

tgtcatttga tcagtttttt ctatagagac tgactggcga ccggcgttct gttgcgctaa     3420

tattttcgca aacgaacatc aatgttcaca ttcgatgtga acagcctcag gaggccttgc     3480

ccgtgtccca gccagtttcg tcccagcccc caacagccaa ctgctatgac cttgccgtaa     3540

tcggcggtgg catcaatggc gtgggcatcg ctgccgacgc cgctggtcgc ggcttgaagg     3600

tcttcctttg cgaaaaagac gacctggccc aacacacgtc gtcagccagc agcaaactga     3660

ttcacggtgg cctgcgctac ctggagcact acgaatttcg tcttgtacgc gaggccctgg     3720

ctgaacgcga agtgctgctg gcgaaggcgc cccacatcgt caaacctatg cgcttcgtgc     3780

ttccgcaccg cccgcacctg cgccctgcat ggatgatccg cgccggcctg ttcctctacg     3840

accacctggg caagcgcaag cgccttggcg cctcgcgcag cctgcgcttt ggcccgggct     3900

acccgctgaa gccggcaatc acccgcggct tcgaatacgc cgactgcgct gtcgatgacg     3960

cccgcctggt agtactcaac gccatggcgg cccgtgagct tggcgcccat atccgcaccc     4020

gcacccgctg cttgcgcgcc gaacgtgtag agggcttgtg gcaggtagag ctacagcacg     4080

ccgatggcag cctccagacg atccatgccc gcgcgctggt caacgccgcc ggcccgtggg     4140

tggccagctt catcaaggac gacctcaagc tcgatgcgcc ttacggcatc cgcctgatcc     4200

agggcagcca cattatcgtg ccgcgcctgt atgaaggcga acatgcctac attctgcaga     4260

atgaagacca gcgcatcgtc ttctgcatcc cgtatctgga ccgctttacc ctgatcggca     4320

ccactgaccg ggaatacagc ggcgacccgg ccgcggtcgc cattaccgag caggaaacac     4380

attacctgtt gaaggtcgtc aacgagcact tcaaccacca actgagccaa gccgacatcc     4440

tgcataccta ctccggcgtt cggccgctgt gcaacgacga gtcggacaac ccttcggcgg     4500

tgacccggga ctacacactc gccctttcgg cagaggttgg ccaggcaccg ctgctgtcgg     4560

tgttcggcgg caagctcacc acctaccgca agctggccga atcggccatg gctgaactca     4620

agcccttctt cactcagatg cgcgccccct ggaccgccag tgcgccacta ccgggcgcag     4680

agggcatgac aacggtacag gcgctgatcg atgccgtact ggcacgttgc ggctggttgc     4740

ctgtcgacct ggccaaacgc tgggtgctca cttacggcag ccgcgtgtgg cggctgctcg     4800

atggcgttca cggccccgaa gacctcgggc aggctatcgg ggccgggctg ttcacccgcg     4860

aggtggatta cctgctggag gaggaatggg cagagcaaac cgctgacatt atctggcgtc     4920

gcaccaagct gggcctgtcg ctgaccgcgc aggagcagat ggccctggag gcgtatctgc     4980

aacaagcccg caaggtgcgg gcgaatagcc gcgcagcctg a                         5021


<210> 2
<211> 756
<212> DNA
<213> Pseudomonas putida KT2440

<220> 
<221> source
<222> 1..756
<223> /mol_type="DNA"
      /note="Secuencia nucleotidica del gen glpR en Pseudomonas putida 
      KT2440"
      /organism="Pseudomonas putida KT2440"

<400> 2
atgaatctgc ccccccgcca acaacaaatc ctcgaactgg tacgcgaacg cggctatgtc      60

agtatcgagg aaatggcgca gctgttcgtt gtcactcccc agaccatccg acgcgacatc     120

aatcagcttg ccgagctcaa cctgctgcgc cgttaccacg gcggcgcagc ttacgactcg     180

agcatcgaga acaccgccta cgccatgcgt gccgaccaga tgcgtgacga aaagcagcgc     240

attgccgacg ccgtagcccg gcagattccc gaccatgcgt cgctgttcat caacatcggc     300

accaccaccg aatcgatcgc acgggcgctg ctcaaccaca gccacctgaa agtcatcacc     360

aataacttgc atgtggcggc gatcctggcc gccaaggacg atttcgaagt gctggtggca     420

ggtggcacgg tgcgccgaga tggtggtgtc gtcggccagg ccagtgtcga tttcatcaac     480

cagttcaagg tcgacttcgc cctggtgggc atcagcggta tcgacgagga tggcagcctg     540

ctcgatttcg actatcagga agtgcgggtg tcgcaagcca tcattgccaa tgcccggcaa     600

gtgatcctcg ccgccgactc cagcaagttc gggcgcaatg ccatggtgcg cctgggctcg     660

atcgacctgg tcgactgcct ggtgaccgac cggacaccaa ccccggccct tacccaattg     720

ttgaaccaga acaagatcag gcttgaggtg gtctga                               756


<210> 3
<211> 833
<212> DNA
<213> Pseudomonas putida KT2440

<220> 
<221> source
<222> 1..833
<223> /mol_type="DNA"
      /note="Secuencia nucleotidica de la zona localizada a 5'del gen g
      lpR que ha sido clonada para construir el vector pK18mobsacB-GlpR
      "
      /organism="Pseudomonas putida KT2440"

<400> 3
ggccaagcca agaataccta cggcactggc tgcttcctgt tgatgaacac gggcgaccag      60

gcggtgaagt cgtcccacgg cctgctcacg accattgcct gcggcccgcg tggtgaggtg     120

gcctatgcac tggaaggcgc ggtattcaat ggtggctcta ccgtgcaatg gctgcgtgac     180

gaactgaaga tcgtcaacga cgccctggat accgagtatt tcgccagcaa ggtcaaggac     240

agcaatggcg tgtacctggt gccagccttt accggcctgg gtgccccgta ctgggacccg     300

tatgcccgtg gtgcgttgtt cggcctgaca cgtggcgtga aggtagacca catcatccgc     360

gccgcactgg agtccatcgc ctaccagacc cgcgatgtcc tcgacgccat gcagcaggac     420

tgtggccagc gcctgtccga gctgcgcgtg gacggtggtg ccgtggccaa caacttcctc     480

atgcagttcc aggccgacat cctcggcacc tgcgtggagc ggccgcagat gcgcgaaacc     540

acggcgctgg gcgccgccta tctggcaggc ctggcctgtg gcttctggag tggcctggac     600

gagctgcgcg acaaggcgat catcgagcgt gagttcagcc cgcaactgga cgaaacgcag     660

aaagaaaagc tgtatgccgg ctggaagaag gcggtcgagc gtacccgtga ctgggaagat     720

cacgaggcat gatagctgtt ggggccgcca tgcggcccca atcgtctact ggccgtcatc     780

cccttcctac ggcatcattg cagaatttgt ccggcagccc caaaggaccg ccc            833


<210> 4
<211> 793
<212> DNA
<213> Pseudomonas putida KT2440

<220> 
<221> source
<222> 1..793
<223> /mol_type="DNA"
      /note="Secuencia nucleotidica de la zona localizada a 3' del gen 
      glpR que ha sido clonada para construir el vector pK18mobsacB-Glp
      R"
      /organism="Pseudomonas putida KT2440"

<400> 4
gggctggtgg gtgcatgccg ccagactttt ggtgtctgcg tgagcaagcg ccccctcacc      60

caagccgtac cccctgaatt ttcgtaaatg ttcacttcat tgtcatttga tcagtttttt     120

ctatagagac tgactggcga ccggcgttct gttgcgctaa tattttcgca aacgaacatc     180

aatgttcaca ttcgatgtga acagcctcag gaggccttgc ccgtgtccca gccagtttcg     240

tcccagcccc caacagccaa ctgctatgac cttgccgtaa tcggcggtgg catcaatggc     300

gtgggcatcg ctgccgacgc cgctggtcgc ggcttgaagg tcttcctttg cgaaaaagac     360

gacctggccc aacacacgtc gtcagccagc agcaaactga ttcacggtgg cctgcgctac     420

ctggagcact acgaatttcg tcttgtacgc gaggccctgg ctgaacgcga agtgctgctg     480

gcgaaggcgc cccacatcgt caaacctatg cgcttcgtgc ttccgcaccg cccgcacctg     540

cgccctgcat ggatgatccg cgccggcctg ttcctctacg accacctggg caagcgcaag     600

cgccttggcg cctcgcgcag cctgcgcttt ggcccgggct acccgctgaa gccggcaatc     660

acccgcggct tcgaatacgc cgactgcgct gtcgatgacg cccgcctggt agtactcaac     720

gccatggcgg cccgtgagct tggcgcccat atccgcaccc gcacccgctg cttgcgcgcc     780

gaacgtgtag agg                                                        793


<210> 5
<211> 31
<212> DNA
<213> Pseudomonas putida KT2440

<220> 
<221> source
<222> 1..31
<223> /mol_type="DNA"
      /note="Oligonucleotido  FR1 - GlpR - 5´ "
      /organism="Pseudomonas putida KT2440"

<400> 5
gctctagagg ccaagccaag aatacctacg g                                   31


<210> 6
<211> 27
<212> DNA
<213> Pseudomonas putida KT2440

<220> 
<221> source
<222> 1..27
<223> /mol_type="DNA"
      /note="Oligonucleotido - FR1 - GlpR - 3´"
      /organism="Pseudomonas putida KT2440"

<400> 6
cgggatccgg gcggtccttt ggggctg                                        27


<210> 7
<211> 26
<212> DNA
<213> Pseudomonas putida KT2440

<220> 
<221> source
<222> 1..26
<223> /mol_type="DNA"
      /note="Oligonucleotido - FR2 - GlpR - 5´"
      /organism="Pseudomonas putida KT2440"

<400> 7
cgggatccgg gctggtgggt gcatgc                                         26


<210> 8
<211> 28
<212> DNA
<213> Pseudomonas putida KT2440

<220> 
<221> source
<222> 1..28
<223> /mol_type="DNA"
      /note="Oligonucleotido FR2 - GlpR - 3´"
      /organism="Pseudomonas putida KT2440"

<400> 8
cccaagcttc ctctacacgt tcggcgcg                                       28



