               SEQUENCE LISTING

<110> DSM IP Assets B.V.

<120> PROCESS FOR ENZYMATIC HYDROLYSIS OF LIGNOCELLULOSIC MATERIAL AND FERMENTATION OF SUGARS

<130> 31238-WO-PCT

<150>  EPEP15173120.5 
<151>  2015-06-22

<160> 11

<170> BiSSAP 1.2

<210> 1
<211> 548
<212> PRT
<213> Escherichia coli


<400> 1
Met Ser Asn Lys Pro Phe Ile Tyr Gln Ala Pro Phe Pro Met Gly Lys 
1               5                   10                  15      
Asp Asn Thr Glu Tyr Tyr Leu Leu Thr Ser Asp Tyr Val Ser Val Ala 
            20                  25                  30          
Asp Phe Asp Gly Glu Thr Ile Leu Lys Val Glu Pro Glu Ala Leu Thr 
        35                  40                  45              
Leu Leu Ala Gln Gln Ala Phe His Asp Ala Ser Phe Met Leu Arg Pro 
    50                  55                  60                  
Ala His Gln Lys Gln Val Ala Ala Ile Leu His Asp Pro Glu Ala Ser 
65                  70                  75                  80  
Glu Asn Asp Lys Tyr Val Ala Leu Gln Phe Leu Arg Asn Ser Glu Ile 
                85                  90                  95      
Ala Ala Lys Gly Val Leu Pro Thr Cys Gln Asp Thr Gly Thr Ala Ile 
            100                 105                 110         
Ile Val Gly Lys Lys Gly Gln Arg Val Trp Thr Gly Gly Gly Asp Glu 
        115                 120                 125             
Glu Thr Leu Ser Lys Gly Val Tyr Asn Thr Tyr Ile Glu Asp Asn Leu 
    130                 135                 140                 
Arg Tyr Ser Gln Asn Ala Ala Leu Asp Met Tyr Lys Glu Val Asn Thr 
145                 150                 155                 160 
Gly Thr Asn Leu Pro Ala Gln Ile Asp Leu Tyr Ala Val Asp Gly Asp 
                165                 170                 175     
Glu Tyr Lys Phe Leu Cys Val Ala Lys Gly Gly Gly Ser Ala Asn Lys 
            180                 185                 190         
Thr Tyr Leu Tyr Gln Glu Thr Lys Ala Leu Leu Thr Pro Gly Lys Leu 
        195                 200                 205             
Lys Asn Phe Leu Val Glu Lys Met Arg Thr Leu Gly Thr Ala Ala Cys 
    210                 215                 220                 
Pro Pro Tyr His Ile Ala Phe Val Ile Gly Gly Thr Ser Ala Glu Thr 
225                 230                 235                 240 
Asn Leu Lys Thr Val Lys Leu Ala Ser Ala His Tyr Tyr Asp Glu Leu 
                245                 250                 255     
Pro Thr Glu Gly Asn Glu His Gly Gln Ala Phe Arg Asp Val Gln Leu 
            260                 265                 270         
Glu Gln Glu Leu Leu Glu Glu Ala Gln Lys Leu Gly Leu Gly Ala Gln 
        275                 280                 285             
Phe Gly Gly Lys Tyr Phe Ala His Asp Ile Arg Val Ile Arg Leu Pro 
    290                 295                 300                 
Arg His Gly Ala Ser Cys Pro Val Gly Met Gly Val Ser Cys Ser Ala 
305                 310                 315                 320 
Asp Arg Asn Ile Lys Ala Lys Ile Asn Arg Glu Gly Ile Trp Ile Glu 
                325                 330                 335     
Lys Leu Glu His Asn Pro Gly Gln Tyr Ile Pro Gln Glu Leu Arg Gln 
            340                 345                 350         
Ala Gly Glu Gly Glu Ala Val Lys Val Asp Leu Asn Arg Pro Met Lys 
        355                 360                 365             
Glu Ile Leu Ala Gln Leu Ser Gln Tyr Pro Val Ser Thr Arg Leu Ser 
    370                 375                 380                 
Leu Thr Gly Thr Ile Ile Val Gly Arg Asp Ile Ala His Ala Lys Leu 
385                 390                 395                 400 
Lys Glu Leu Ile Asp Ala Gly Lys Glu Leu Pro Gln Tyr Ile Lys Asp 
                405                 410                 415     
His Pro Ile Tyr Tyr Ala Gly Pro Ala Lys Thr Pro Ala Gly Tyr Pro 
            420                 425                 430         
Ser Gly Ser Leu Gly Pro Thr Thr Ala Gly Arg Met Asp Ser Tyr Val 
        435                 440                 445             
Asp Leu Leu Gln Ser His Gly Gly Ser Met Ile Met Leu Ala Lys Gly 
    450                 455                 460                 
Asn Arg Ser Gln Gln Val Thr Asp Ala Cys His Lys His Gly Gly Phe 
465                 470                 475                 480 
Tyr Leu Gly Ser Ile Gly Gly Pro Ala Ala Val Leu Ala Gln Gln Ser 
                485                 490                 495     
Ile Lys His Leu Glu Cys Val Ala Tyr Pro Glu Leu Gly Met Glu Ala 
            500                 505                 510         
Ile Trp Lys Ile Glu Val Glu Asp Phe Pro Ala Phe Ile Leu Val Asp 
        515                 520                 525             
Asp Lys Gly Asn Asp Phe Phe Gln Gln Ile Val Asn Lys Gln Cys Ala 
    530                 535                 540                 
Asn Cys Thr Lys 
545             

<210> 2
<211> 8074
<212> DNA
<213> Artificial sequence

<220> 
<221> source
<222> 1..8074
<223> /organism="Artificial sequence"
      /note="Sequence of plasmid pSUC223"
      /mol_type="unassigned DNA"

<400> 2
gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca       60

cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct      120

cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat      180

tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttaa      240

ttaaaggagg tgcacgcatt atggagacca ctacgatacg atagctgcgt tgttgttgaa      300

ggggtttctt aaggttgttt tcgttgaagg taaatattgg tcgtttttgt gcagcatatt      360

gtcctctaga tgcaaactct gcaggtccat ttgcagtaaa gtgagttgcc tctcgaagaa      420

tcattaattt cgtataaccg tcactattaa agtcagaaaa taaattctgt cgtagacaat      480

gttaccataa tgttcttgtc cattttgcat acactttaaa tattcatttg atttctcagg      540

gttcatgatc ataataaatt gcgcattcgc aaggcggtag tattataatg gggtccatca      600

ttctgtagca agaagttaca gtacgctgtt caagcgttaa acaagataag taatctcgaa      660

tgaaacattc atatttcgca tgagccaaca tacagttgct gagtaatctt cattgcgctt      720

atttatcggc attgagattg taaaggaagt aaaacgcatt tttgcagatc tgttctctta      780

tgtattttta atcgtccttg tatggaagta tcaaagggga cgttcttcac ctccttggaa      840

ggatcccagc gccagtaggg ttgttgagct tagtaaaaat gtgcgcacca caagcctaca      900

tgactccacg tcacatgaaa ccacaccgtg gggccttgtt gcgctaggaa taggatatgc      960

gacgaagacg cttctgctta gtaaccacac cacattttca gggggtcgat ctgcttgctt     1020

cctttactgt cacgagcggc ccataatcgc gctttttttt taaaaggcgc gagacagcaa     1080

acaggaagct cgggtttcaa ccttcggagt ggtcgcagat ctggagactg gatctttaca     1140

atacagtaag gcaagccacc atctgcttct taggtgcatg cgacggtatc cacgtgcaga     1200

acaacatagt ctgaagaagg gggggaggag catgttcatt ctctgtagca gtaagagctt     1260

ggtgataatg accaaaactg gagtctcgaa atcatataaa tagacaatat attttcacac     1320

aatgagattt gtagtacagt tctattctct ctcttgcata aataagaaat tcatcaagaa     1380

cttggtttga tatttcacca acacacacaa aaaacagtac ttcactaaat ttacacacaa     1440

aacaaaatgt ccaacaagcc tttcatctac caagctccat tcccaatggg taaggacaac     1500

actgaatact acttgttgac ttctgactac gtttccgttg ctgatttcga tggtgaaacc     1560

atcttgaagg ttgaaccaga agccttgact ttgttggctc aacaagcctt ccacgatgct     1620

tctttcatgt tgcgtccagc tcaccaaaag caagttgctg ccattttgca cgacccagaa     1680

gcctccgaaa acgacaaata cgttgctttg caattcttga gaaactctga aattgctgcc     1740

aagggtgtct taccaacttg tcaagacact ggtactgcca tcattgtcgg taagaagggt     1800

caaagagtct ggaccggtgg tggtgacgaa gaaactctat ccaagggtgt ttacaacact     1860

tacattgaag ataatttacg ttactctcaa aatgctgctt tggacatgta caaggaagtc     1920

aacactggta ccaacttgcc agctcaaatc gacttatacg ctgttgacgg tgacgaatac     1980

aagttcttgt gtgttgccaa gggtggtggt tctgctaaca agacctactt gtaccaagaa     2040

accaaggctt tgttgactcc aggtaaattg aagaacttct tggtcgaaaa gatgagaact     2100

ttgggtactg ctgcttgtcc accataccac attgctttcg ttatcggtgg tacttccgct     2160

gaaaccaact tgaaaaccgt caaattggct tccgctcact actacgatga attgccaact     2220

gaaggtaacg aacacggtca agccttcaga gatgtccaat tggaacaaga attgttggaa     2280

gaagctcaaa aattaggttt gggtgctcaa tttggtggta aatactttgc tcacgatatc     2340

agagttatca gattaccaag acatggtgct tcttgtccag ttggtatggg tgtttcctgt     2400

tctgctgaca gaaacatcaa ggccaagatc aacagagaag gtatctggat tgaaaaattg     2460

gaacacaacc caggtcaata catcccacaa gaattgagac aagctggtga aggtgaagct     2520

gtcaaggttg acttgaacag accaatgaag gaaatcttgg ctcaattatc tcaataccca     2580

gtttccacca gattatcttt gaccggtacc atcattgtcg gtcgtgacat tgctcatgcc     2640

aagttgaagg aattgattga tgctggtaag gaattgcctc aatacatcaa ggaccatcca     2700

atctactacg ctggtccagc caagacccca gctggttacc catctggttc tttgggtcca     2760

accaccgctg gtagaatgga ctcttacgtt gacttgctac aatctcacgg tggttccatg     2820

atcatgttgg ctaagggtaa cagatctcaa caagtcaccg atgcttgtca caagcacggt     2880

ggtttctatt tgggttccat tggtggtcca gctgctgtct tggctcaaca atctatcaag     2940

cacttggaat gtgttgctta cccagaattg ggtatggaag ccatctggaa gattgaagtc     3000

gaagatttcc cagctttcat cttagtcgat gacaagggta acgacttctt ccaacaaatt     3060

gtcaacaagc aatgtgccaa ctgtaccaag taagcgtacg ataaagcaat cttgatgagg     3120

ataatgattt ttttttgaat atacataaat actaccgttt ttctgctaga ttttgtgaag     3180

acgtaaataa gtacatatta ctttttaagc caagacaaga ttaagcatta actttaccct     3240

tttctcttct aagtttcaat actagttatc actgtttaaa agttatggcg agaacgtcgg     3300

cggttaaaat atattaccct gaacgtggtg aattgaagtt ctaggatggt ttaaagattt     3360

ttcctttttg ggaaataagt aaacaatata ttgctgcctt ggcgcgcccc ttccgggccc     3420

acgcgtggcc ggccgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acgaattcta     3480

ccgttcgtat aatgtatgct atacgaagtt atagatctgt ttagcttgcc tcgtccccgc     3540

cgggtcaccc ggccagcgac atggaggccc agaataccct ccttgacagt cttgacgtgc     3600

gcagctcagg ggcatgatgt gactgtcgcc cgtacattta gcccatacat ccccatgtat     3660

aatcatttgc atccatacat tttgatggcc gcacggcgcg aagcaaaaat tacggctcct     3720

cgctgcagac ctgcgagcag ggaaacgctc ccctcacaga cgcgttgaat tgtccccacg     3780

ccgcgcccct gtagagaaat ataaaaggtt aggatttgcc actgaggttc ttctttcata     3840

tacttccttt taaaatcttg ctaggataca gttctcacat cacatccgaa cataaacaac     3900

catgggtaag gaaaagactc acgtttcgag gccgcgatta aattccaaca tggatgctga     3960

tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg     4020

attgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc     4080

caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc     4140

gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc     4200

cggcaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga     4260

tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa     4320

cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga     4380

tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat     4440

gcataagctt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga     4500

taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat     4560

cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc     4620

attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca     4680

gtttcatttg atgctcgatg agtttttcta atcagtactg acaataaaaa gattcttgtt     4740

ttcaagaact tgtcatttgt atagtttttt tatattgtag ttgttctatt ttaatcaaat     4800

gttagcgtga tttatatttt ttttcgcctc gacatcatct gcccagatgc gaagttaagt     4860

gcgcagaaag taatatcatg cgtcaatcgt atgtgaatgc tggtcgctat actgctgtcg     4920

attcgatact aacgccgcca tccagtgtcg aaaacgagct cataacttcg tataatgtat     4980

gctatacgaa cggtagaatt cgaatcagat ccactagtgg cctatgcggc cgcgatggga     5040

cgtcagcact gtacttgttt ttgcgactag attgtaaatc attctttatt taatctcttt     5100

ctttaactac tgcttaaagt ataatttggt ccgtagttta ataactatac taagcgtaac     5160

aatgcatact gacattataa gcctgaacat tacgagttta agttgtatgt aggcgttctg     5220

taagaggtta ctgcgtaaat tatcaacgaa tgcattggtg tatttgcgaa agctacttct     5280

tttaacaagt atttacataa gaataatggt gatctgctca actgatttgg tgataactct     5340

aactttttta gcaacaattt aaaagataat tcgaacatat ataacagtag gaagaatttg     5400

tgtacgtcaa attaagataa tttagcatta ccaaagttat taacctaaac ataaaatata     5460

tatgagacac atgtggaaat cgtatgaaac aactgttatg aaactgacaa gaatgaatat     5520

atagagtaag ctccgcttgt aaagaggaat cacttaagtg tataaatgtc tcgacgatta     5580

ctttagatcc aagattgatg attgatatta ctctgtaata cttaagctct tttaatagct     5640

cactgttgta ttacgggctc gagtaatacc ggaattcact ggccgtcgtt ttacaacgtc     5700

gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg     5760

ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc     5820

tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac     5880

accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc     5940

gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt     6000

acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac     6060

cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga     6120

taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta     6180

tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat     6240

aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc     6300

ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga     6360

aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca     6420

acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt     6480

ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg     6540

gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc     6600

atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata     6660

acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt     6720

tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag     6780

ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca     6840

aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg     6900

aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg     6960

ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag     7020

atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg     7080

aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag     7140

accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga     7200

tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt     7260

tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc     7320

tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc     7380

cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac     7440

caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac     7500

cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt     7560

cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct     7620

gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat     7680

acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt     7740

atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg     7800

cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt     7860

gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt     7920

tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg     7980

tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg     8040

agcgcagcga gtcagtgagc gaggaagcgg aaga                                 8074


<210> 3
<211> 452
<212> PRT
<213> Artificial sequence

<220> 
<223> Modified protein sequence of YEL047c encoding FRD1 (E.C. 1.3.1.6,
      UniProt accession number P32614)

<400> 3
Met Asn Glu Leu Val Asn Lys Tyr Asn Ile Pro Val Thr Ile Leu Glu 
1               5                   10                  15      
Lys Ala Ser Ser Ile Gly Gly Asn Ser Ile Lys Ala Ser Ser Gly Ile 
            20                  25                  30          
Asn Gly Ala Cys Thr Glu Thr Gln Arg His Phe His Ile Glu Asp Ser 
        35                  40                  45              
Pro Arg Leu Phe Glu Asp Asp Thr Ile Lys Ser Ala Lys Gly Lys Gly 
    50                  55                  60                  
Val Gln Glu Leu Met Ala Lys Leu Ala Asn Asp Ser Pro Leu Ala Ile 
65                  70                  75                  80  
Glu Trp Leu Lys Asn Glu Phe Asp Leu Lys Leu Asp Leu Leu Ala Gln 
                85                  90                  95      
Leu Gly Gly His Ser Val Ala Arg Thr His Arg Ser Ser Gly Lys Leu 
            100                 105                 110         
Pro Pro Gly Phe Glu Ile Val Ser Ala Leu Ser Asn Asn Leu Lys Lys 
        115                 120                 125             
Leu Ala Glu Thr Lys Pro Glu Leu Val Lys Ile Asn Leu Asp Ser Lys 
    130                 135                 140                 
Val Val Asp Ile His Glu Lys Asp Gly Ser Ile Ser Ala Val Val Tyr 
145                 150                 155                 160 
Glu Asp Lys Asn Gly Glu Lys His Met Val Ser Ala Asn Asp Val Val 
                165                 170                 175     
Phe Cys Ser Gly Gly Phe Gly Phe Ser Lys Glu Met Leu Lys Glu Tyr 
            180                 185                 190         
Ala Pro Glu Leu Val Asn Leu Pro Thr Thr Asn Gly Gln Gln Thr Thr 
        195                 200                 205             
Gly Asp Gly Gln Arg Leu Leu Gln Lys Leu Gly Ala Asp Leu Ile Asp 
    210                 215                 220                 
Met Asp Gln Ile Gln Val His Pro Thr Gly Phe Ile Asp Pro Asn Asp 
225                 230                 235                 240 
Arg Ser Ser Ser Trp Lys Phe Leu Ala Ala Glu Ser Leu Arg Gly Leu 
                245                 250                 255     
Gly Gly Ile Leu Leu Asn Pro Ile Thr Gly Arg Arg Phe Val Asn Glu 
            260                 265                 270         
Leu Thr Thr Arg Asp Val Val Thr Ala Ala Ile Gln Lys Val Cys Pro 
        275                 280                 285             
Gln Glu Asp Asn Arg Ala Leu Leu Val Met Gly Glu Lys Met Tyr Thr 
    290                 295                 300                 
Asp Leu Lys Asn Asn Leu Asp Phe Tyr Met Phe Lys Lys Leu Val Gln 
305                 310                 315                 320 
Lys Leu Thr Leu Ser Gln Val Val Ser Glu Tyr Asn Leu Pro Ile Thr 
                325                 330                 335     
Val Ala Gln Leu Cys Glu Glu Leu Gln Thr Tyr Ser Ser Phe Thr Thr 
            340                 345                 350         
Lys Ala Asp Pro Leu Gly Arg Thr Val Ile Leu Asn Glu Phe Gly Ser 
        355                 360                 365             
Asp Val Thr Pro Glu Thr Val Val Phe Ile Gly Glu Val Thr Pro Val 
    370                 375                 380                 
Val His Phe Thr Met Gly Gly Ala Arg Ile Asn Val Lys Ala Gln Val 
385                 390                 395                 400 
Ile Gly Lys Asn Asp Glu Arg Leu Leu Lys Gly Leu Tyr Ala Ala Gly 
                405                 410                 415     
Glu Val Ser Gly Gly Val His Gly Ala Asn Arg Leu Gly Gly Ser Ser 
            420                 425                 430         
Leu Leu Glu Cys Val Val Phe Gly Arg Thr Ala Ala Glu Ser Ile Ala 
        435                 440                 445             
Asn Asp Arg Lys 
    450         

<210> 4
<211> 2260
<212> DNA
<213> Artificial sequence

<220> 
<221> source
<222> 1..2260
<223> /organism="Artificial sequence"
      /note="Synthetic TDH3p-FRD1-TDH3t gene"
      /mol_type="unassigned DNA"

<400> 4
ttagtcaaaa aattagcctt ttaattctgc tgtaacccgt acatgcccaa aatagggggc      60

gggttacaca gaatatataa catcgtaggt gtctgggtga acagtttatt cctggcatcc     120

actaaatata atggagcccg ctttttaagc tggcatccag aaaaaaaaag aatcccagca     180

ccaaaatatt gttttcttca ccaaccatca gttcataggt ccattctctt agcgcaacta     240

cagagaacag gggcacaaac aggcaaaaaa cgggcacaac ctcaatggag tgatgcaacc     300

tgcctggagt aaatgatgac acaaggcaat tgacccacgc atgtatctat ctcattttct     360

tacaccttct attaccttct gctctctctg atttggaaaa agctgaaaaa aaaggttgaa     420

accagttccc tgaaattatt cccctacttg actaataagt atataaagac ggtaggtatt     480

gattgtaatt ctgtaaatct atttcttaaa cttcttaaat tctactttta tagttagtct     540

tttttttagt tttaaaacac caagaactta gtttcgaata aacacacata aacaaacaaa     600

atgaacgaat tagtcaacaa atacaacatt ccagtcacca ttttggaaaa ggcttcttcc     660

attggtggta actccatcaa ggcttcctcc ggtatcaacg gtgcttgtac cgaaactcaa     720

agacatttcc acattgaaga ttctccaaga ttgtttgaag atgacaccat caaatctgcc     780

aagggtaagg gtgtccaaga attgatggcc aagttggcta acgactctcc attggccatt     840

gaatggttga aaaatgaatt cgacttgaaa ttggacttgt tggctcaatt aggtggtcac     900

tccgttgcca gaactcaccg ttcttctggt aagttgccac caggtttcga aattgtttct     960

gctttgtcca acaacttgaa gaaattggct gaaaccaagc cagaattggt caagatcaac    1020

ttggactcca aggttgtcga tatccacgaa aaggacggtt ccatctctgc tgttgtttac    1080

gaagataaga acggtgaaaa gcacatggtt tctgctaacg atgttgtctt ttgttctggt    1140

ggtttcggtt tctccaagga aatgttgaag gaatacgctc cagaattggt caacttgcca    1200

accaccaacg gtcaacaaac cactggtgat ggtcaaagat tactacaaaa gttgggtgct    1260

gacttgattg acatggacca aatccaagtt cacccaactg gtttcattga cccaaacgac    1320

cgttcttctt cctggaaatt cttggctgct gaatctttga gaggtttggg tggtatcttg    1380

ttgaacccaa tcactggtag aagatttgtc aatgaattga ccaccagaga tgtcgttacc    1440

gctgccatcc aaaaggtctg tccacaagaa gataacagag ctttgttggt catgggtgaa    1500

aagatgtaca ctgacttgaa gaacaacttg gacttctaca tgttcaagaa gttggttcaa    1560

aagttgactt tatctcaagt tgtttctgaa tacaacttgc caatcactgt tgctcaatta    1620

tgtgaagaat tgcaaactta ctcctctttc accaccaagg ctgacccttt gggtagaacc    1680

gttatcttga acgaattcgg ttctgatgtc actccagaaa ctgttgtttt catcggtgaa    1740

gtcactccag ttgtccactt caccatgggt ggtgccagaa tcaacgtcaa ggctcaagtt    1800

atcggtaaga acgatgaaag attattgaag ggtttatacg ctgctggtga agtctctggt    1860

ggtgtccacg gtgctaacag attaggtggt tcctctctat tggaatgtgt tgttttcggt    1920

agaactgctg ctgaatccat tgccaacgac cgcaagtaag gtgaatttac tttaaatctt    1980

gcatttaaat aaattttctt tttatagctt tatgacttag tttcaattta tatactattt    2040

taatgacatt ttcgattcat tgattgaaag ctttgtgttt tttcttgatg cgctattgca    2100

ttgttcttgt ctttttcgcc acatgtaata tctgtagtag atacctgata cattgtggat    2160

gctgagtgaa attttagtta ataatggagg cgctcttaat aattttgggg atattggctt    2220

ttttttttaa agtttacaaa tgaatttttt ccgccaggat                          2260


<210> 5
<211> 84
<212> DNA
<213> Artificial sequence

<220> 
<221> source
<222> 1..84
<223> /organism="Artificial sequence"
      /note="Primer 1 (TDH3p FW with PDC6 5' overhang)"
      /mol_type="unassigned DNA"

<400> 5
gtcctctttt atatacagta taaataaaaa acccacgtaa tatagcaaaa acatattgcc     60

aacaaattag tcaaaaaatt agcc                                            84


<210> 6
<211> 82
<212> DNA
<213> Artificial sequence

<220> 
<221> source
<222> 1..82
<223> /organism="Artificial sequence"
      /note="Primer 2 (TDH3t REV with overhang to pSUC228 Cre-1)"
      /mol_type="unassigned DNA"

<400> 6
cgctatactg atttaaatat aacttcgtat agcatacatt atacgaacgg tactcgaggt     60

cgaatcctgg cggaaaaaat tc                                              82


<210> 7
<211> 86
<212> DNA
<213> Artificial sequence

<220> 
<221> source
<222> 1..86
<223> /organism="Artificial sequence"
      /note="Primer 3 (pSUC228 Cre-1 FW with overhang TDH3t)"
      /mol_type="unassigned DNA"

<400> 7
cttaataatt ttggggatat tggctttttt ttttaaagtt tacaaatgaa ttttttccgc     60

caggattcga cctcgagtac cgttcg                                          86


<210> 8
<211> 21
<212> DNA
<213> Artificial sequence

<220> 
<221> source
<222> 1..21
<223> /organism="Artificial sequence"
      /note="Primer 4 (DBC-03373, pSUC228 Cre-1 REV)"
      /mol_type="unassigned DNA"

<400> 8
gcaatttcgg ctatacgtaa c                                              21


<210> 9
<211> 4780
<212> DNA
<213> Artificial sequence

<220> 
<221> source
<222> 1..4780
<223> /organism="Artificial sequence"
      /note="Sequence of plasmid pSUC228"
      /mol_type="unassigned DNA"

<400> 9
aaatagcttg ccttgtcccc gccgggtcac ccggccagcg acatggaggc ccagaatacc       60

ctccttgaca gtcttgacgt gcgcagctca ggggcatgat gtgactgtcg cccgtacatt      120

tagcccatac atccccatgt ataatcattt gcatccatac attttgatgg ccgcacggcg      180

cgaagcaaaa attacggctc ctcgctgcag acctgcgagc agggaaacgc tcccctcaca      240

gacgcgttga attgtcccca cgccgcgccc ctgtagagaa atataaaagg ttaggatttg      300

ccactgaggt tcttctttca tatacttcct tttaaaatct tgctaggata cagttctcac      360

atcacatccg aacataaaca accatgggta ccactcttga cgacacggct taccggtacc      420

gcaccagtgt cccgggggac gccgaggcca tcgaggcact ggatgggtcc ttcaccaccg      480

acaccgtctt ccgcgtcacc gccaccgggg acggcttcac cctgcgggag gtgccggtgg      540

acccgcccct gaccaaggtg ttccccgacg acgaatcgga cgacgaatcg gacgacgggg      600

aggacggcga cccggactcc cggacgttcg tcgcgtacgg ggacgacggc gacctggcgg      660

gcttcgtggt cgtctcgtac tccggctgga accgccggct gaccgtcgag gacatcgagg      720

tcgccccgga gcaccggggg cacggggtcg ggcgcgcgtt gatggggctc gcgacggagt      780

tcgcccgcga gcggggcgcc gggcacctct ggctggaggt caccaacgtc aacgcaccgg      840

cgatccacgc gtaccggcgg atggggttca ccctctgcgg cctggacacc gccctgtacg      900

acggcaccgc ctcggacggc gagcaggcgc tctacatgag catgccctgc ccctaatcag      960

tactgacaat aaaaagattc ttgttttcaa gaacttgtca tttgtatagt ttttttatat     1020

tgtagttgtt ctattttaat caaatgttag cgtgatttat attttttttc gcctcgacat     1080

catctgccca gatgcgaagt taagtgcgca gaaagtaata tcatgcgtca atcgtatgtg     1140

aatgctggtc gctatactgc tgtcgattcg atactaacgc cgccatccag tgtcgattta     1200

aatctagtac ggattagaag ccgccgagcg ggtgacagcc ctccgaagga agactctcct     1260

ccgtgcgtcc tcgtcttcac cggtcgcgtt cctgaaacgc agatgtgcct cgcgccgcac     1320

tgctccgaac aataaagatt ctacaatact agcttttatg gttatgaaga ggaaaaattg     1380

gcagtaacct ggccccacaa accttcaaat gaacgaatca aattaacaac cataggatga     1440

taatgcgatt agttttttag ccttatttct ggggtaatta atcagcgaag cgatgatttt     1500

tgatctatta acagatatat aaatgcaaaa actgcataac cactttaact aatactttca     1560

acattttcgg tttgtattac ttcttattca aatgtaataa aagtatcaac aaaaaattgt     1620

taatatacct ctatacttta acgtcaagga gaaaaaaccc cggattctag aactagtgga     1680

tcccccgggc tgcaggaatt cgatatcaag cttatcgata ccgtcgaggg gcagagccga     1740

tcctgtacac tttacttaaa accattatct gagtgttaaa tgtccaattt actgaccgta     1800

caccaaaatt tgcctgcatt accggtcgat gcaacgagtg atgaggttcg caagaacctg     1860

atggacatgt tcagggatcg ccaggcgttt tctgagcata cctggaaaat gcttctgtcc     1920

gtttgccggt cgtgggcggc atggtgcaag ttgaataacc ggaaatggtt tcccgcagaa     1980

cctgaagatg ttcgcgatta tcttctatat cttcaggcgc gcggtctggc agtaaaaact     2040

atccagcaac atttgggcca gctaaacatg cttcatcgtc ggtccgggct gccacgacca     2100

agtgacagca atgctgtttc actggttatg cggcggatcc gaaaagaaaa cgttgatgcc     2160

ggtgaacgtg caaaacaggc tctagcgttc gaacgcactg atttcgacca ggttcgttca     2220

ctcatggaaa atagcgatcg ctgccaggat atacgtaatc tggcatttct ggggattgct     2280

tataacaccc tgttacgtat agccgaaatt gcggcgcgcc ggtacctctt aattaactgg     2340

cctcatgggc cttccgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc     2400

attaacatgg tcatagctgt ttccttgcgt attgggcgct ctccgcttcc tcgctcactg     2460

actcgctgcg ctcggtcgtt cgggtaaagc ctggggtgcc taatgagcaa aaggccagca     2520

aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc     2580

tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata     2640

aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc     2700

gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc     2760

acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga     2820

accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc     2880

ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag     2940

gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag     3000

aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag     3060

ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca     3120

gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga     3180

cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat     3240

cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga     3300

gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg     3360

tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga     3420

gggcttacca tctggcccca gtgctgcaat gataccgcga gaaccacgct caccggctcc     3480

agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac     3540

tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc     3600

agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc     3660

gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc     3720

catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt     3780

ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc     3840

atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg     3900

tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag     3960

cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat     4020

cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc     4080

atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa     4140

aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta     4200

ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa     4260

aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc     4320

gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa     4380

taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt     4440

ggccgctaca gggcgctccc attcgccatt caggctgcgc aactgttggg aagggcgttt     4500

cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat     4560

taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgagc     4620

gcgacgtaat acgactcact atagggcgaa ttggcggaag gccgtcaagg cctaggcgcg     4680

ccatgagctc ggcgcgccgc ggccgcaggc ctggcgccca gctggggccc gtcgacctcg     4740

agtaccgttc gtataatgta tgctatacga agttatattt                           4780


<210> 10
<211> 21
<212> DNA
<213> Artificial sequence

<220> 
<221> source
<222> 1..21
<223> /organism="Artificial sequence"
      /note="Primer 5 (DBC-03374, pSUC225 Cre-2 FW)"
      /mol_type="unassigned DNA"

<400> 10
cgttcactca tggaaaatag c                                              21


<210> 11
<211> 85
<212> DNA
<213> Artificial sequence

<220> 
<221> source
<222> 1..85
<223> /organism="Artificial sequence"
      /note="Primer 6 (pSUC225 Cre-2 REV with PDC6 3' overhang)"
      /mol_type="unassigned DNA"

<400> 11
gtattcaaac tgtgtaagtt tatttatttg caacaataat tcgtttgagt acactactaa     60

tggccggatc ctaccgttcg tatag                                           85


