                         SEQUENCE LISTING

<110>  Keygene N.V.
 
<120>  METHODS FOR GENERATING MARKER FREE TRANSGENIC PLANTS

<130>  P6005831PCT

<150>  EP 07122354.9
<151>  2007-12-05

<160>  5     

<170>  PatentIn version 3.3

<210>  1
<211>  1650
<212>  DNA
<213>  Agrobacterium tumefaciens

<400>  1
atggatccgt ctagcaatga gaatgtctat gtgggtcgcg gtcacaacat cgaaaatgat     60

gatgacactg accccaggcg ttggaagaag gcgaatatca gttccaacac catctccgat    120

attcagatga cgaatggcga agacgtacaa tcagggagcc ctacccgaac ggaagttgta    180

agcccacgtc tggattatgg atcggtcgac tcctcctcca gcctttattc tggcagcgag    240

cacggaaatc aagctgagat tcaaaaagag ctgtccgtct tgttctcgaa catgtctttg    300

ccaggcaacg atcggcgccc ggacgaatac attctcgtgc atcaaacggg acaagatgct    360

tttactggta ttgccaaagg caacctcgac caaatgccca ccaaggcgga atttaacgcg    420

tgctgccgtc tctacaggga cggagccggt aattactacc cgccacctct cgcattcgac    480

aagattagcg ttccagagca actggaggaa aaatggggga tgatggaggc gaaggaacgt    540

aacaaactgc ggtttcagta caagttggac gtatggaatc atgcgcacgc tgatatgggg    600

atcacgggca cagagatctt ttatcaaaca gataagaaca taaagctcga ccggaattat    660

aaactaagac ctgaagaccg atacgtacaa acagaaaaat acgggcgccg ggaaattcaa    720

aagcgatatc aacacgaact ccaggctggt tcgctgctgc ccgatattat gatcaaaact    780

ccccaaaatg acatccactt cgtgtacagg tttgccggcg acaattacgc caacaaacag    840

ttcagcgagt ttgaacacac cgtcaagcgc aggtatggcg acgagactga gatcaaattg    900

aagtcaaagt caggcattat gcatgactcg aaatatctgg aatcctggga acggggcagt    960

gcggatattc gcttcgcgga attcgttggg gaaaatagag ctcacaatcg gcagtttcca   1020

actgcgacag taaatatggg acagcagcca gacgggcagg gcggtttgac ccgcgaccgt   1080

catgtgagcg ttgacttcct aatgcaaagc gcacccaatt cgccttgggc gcaagctttg   1140

aaaaagggag aactgtggga tcgcgttcag ttgcttgctc gcgacggcaa ccgctatctg   1200

tcgccgccca gattggaata ttctgaccct gcacatttca ccgagttgat gaaccgggtt   1260

ggtttacccg catcgatggg tcggcaaagc catgcggcta gtatcaaatt cgaaaagttt   1320

gacgcgcagg cagcggttat tgtcttaaat ggcccagagt tacgtgacat tcatgacttg   1380

tctcctgaaa aactgcaaaa tttgtccacc aaagatgtca tcgtcgccga tcgcaatgag   1440

aatggtcaga gaactggcac gtacaccagc gtcgcggaat atgagcgctt gcagttaagg   1500

ctgccacccg atgcagcggg ggtgcttggt gaagcaactg acaaatattc acgtgatttc   1560

gttcggccag agccggcgtc gcgtccaatc agtgacagcc gcaggatata cgaaagtcga   1620

ccgcgtagcc aaagcgtcaa cagcttttga                                    1650


<210>  2
<211>  549
<212>  PRT
<213>  Agrobacterium tumefaciens

<400>  2

Met Asp Pro Ser Ser Asn Glu Asn Val Tyr Val Gly Arg Gly His Asn 
1               5                   10                  15      


Ile Glu Asn Asp Asp Asp Thr Asp Pro Arg Arg Trp Lys Lys Ala Asn 
            20                  25                  30          


Ile Ser Ser Asn Thr Ile Ser Asp Ile Gln Met Thr Asn Gly Glu Asp 
        35                  40                  45              


Val Gln Ser Gly Ser Pro Thr Arg Thr Glu Val Val Ser Pro Arg Leu 
    50                  55                  60                  


Asp Tyr Gly Ser Val Asp Ser Ser Ser Ser Leu Tyr Ser Gly Ser Glu 
65                  70                  75                  80  


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


Asn Met Ser Leu Pro Gly Asn Asp Arg Arg Pro Asp Glu Tyr Ile Leu 
            100                 105                 110         


Val His Gln Thr Gly Gln Asp Ala Phe Thr Gly Ile Ala Lys Gly Asn 
        115                 120                 125             


Leu Asp Gln Met Pro Thr Lys Ala Glu Phe Asn Ala Cys Cys Arg Leu 
    130                 135                 140                 


Tyr Arg Asp Gly Ala Gly Asn Tyr Tyr Pro Pro Pro Leu Ala Phe Asp 
145                 150                 155                 160 


Lys Ile Ser Val Pro Glu Gln Leu Glu Glu Lys Trp Gly Met Met Glu 
                165                 170                 175     


Ala Lys Glu Arg Asn Lys Leu Arg Phe Gln Tyr Lys Leu Asp Val Trp 
            180                 185                 190         


Asn His Ala His Ala Asp Met Gly Ile Thr Gly Thr Glu Ile Phe Tyr 
        195                 200                 205             


Gln Thr Asp Lys Asn Ile Lys Leu Asp Arg Asn Tyr Lys Leu Arg Pro 
    210                 215                 220                 


Glu Asp Arg Tyr Val Gln Thr Glu Lys Tyr Gly Arg Arg Glu Ile Gln 
225                 230                 235                 240 


Lys Arg Tyr Gln His Glu Leu Gln Ala Gly Ser Leu Leu Pro Asp Ile 
                245                 250                 255     


Met Ile Lys Thr Pro Gln Asn Asp Ile His Phe Val Tyr Arg Phe Ala 
            260                 265                 270         


Gly Asp Asn Tyr Ala Asn Lys Gln Phe Ser Glu Phe Glu His Thr Val 
        275                 280                 285             


Lys Arg Arg Tyr Gly Asp Glu Thr Glu Ile Lys Leu Lys Ser Lys Ser 
    290                 295                 300                 


Gly Ile Met His Asp Ser Lys Tyr Leu Glu Ser Trp Glu Arg Gly Ser 
305                 310                 315                 320 


Ala Asp Ile Arg Phe Ala Glu Phe Val Gly Glu Asn Arg Ala His Asn 
                325                 330                 335     


Arg Gln Phe Pro Thr Ala Thr Val Asn Met Gly Gln Gln Pro Asp Gly 
            340                 345                 350         


Gln Gly Gly Leu Thr Arg Asp Arg His Val Ser Val Asp Phe Leu Met 
        355                 360                 365             


Gln Ser Ala Pro Asn Ser Pro Trp Ala Gln Ala Leu Lys Lys Gly Glu 
    370                 375                 380                 


Leu Trp Asp Arg Val Gln Leu Leu Ala Arg Asp Gly Asn Arg Tyr Leu 
385                 390                 395                 400 


Ser Pro Pro Arg Leu Glu Tyr Ser Asp Pro Ala His Phe Thr Glu Leu 
                405                 410                 415     


Met Asn Arg Val Gly Leu Pro Ala Ser Met Gly Arg Gln Ser His Ala 
            420                 425                 430         


Ala Ser Ile Lys Phe Glu Lys Phe Asp Ala Gln Ala Ala Val Ile Val 
        435                 440                 445             


Leu Asn Gly Pro Glu Leu Arg Asp Ile His Asp Leu Ser Pro Glu Lys 
    450                 455                 460                 


Leu Gln Asn Leu Ser Thr Lys Asp Val Ile Val Ala Asp Arg Asn Glu 
465                 470                 475                 480 


Asn Gly Gln Arg Thr Gly Thr Tyr Thr Ser Val Ala Glu Tyr Glu Arg 
                485                 490                 495     


Leu Gln Leu Arg Leu Pro Pro Asp Ala Ala Gly Val Leu Gly Glu Ala 
            500                 505                 510         


Thr Asp Lys Tyr Ser Arg Asp Phe Val Arg Pro Glu Pro Ala Ser Arg 
        515                 520                 525             


Pro Ile Ser Asp Ser Arg Arg Ile Tyr Glu Ser Arg Pro Arg Ser Gln 
    530                 535                 540                 


Ser Val Asn Ser Phe 
545                 


<210>  3
<211>  5662
<212>  DNA
<213>  Agrobacterium tumefaciens


<220>
<221>  virE2
<222>  (203)..(1024)

<220>
<221>  stop
<222>  (1025)..(1035)

<220>
<221>  loxP
<222>  (1057)..(1090)

<220>
<221>  R6Kori
<222>  (1111)..(1502)

<220>
<221>  nptII promoter
<222>  (1696)..(1833)

<220>
<221>  nptII gene
<222>  (1863)..(2657)

<220>
<221>  3' end T7
<222>  (2807)..(2935)

<220>
<221>  3' end bgh
<222>  (2950)..(3158)

<220>
<221>  6xhis
<222>  (3181)..(3198)

<220>
<221>  v5 epitope
<222>  (3199)..(3240)

<220>
<221>  stop
<222>  (3289)..(3299)

<220>
<221>  virE2
<222>  (3295)..(4890)

<400>  3
atggccatca tcaagccgca tgtgaacaaa aataggacaa cctcgccgat agagagaccg     60

gagtctctca tagaggaaat gagcggcagt catccgccga gtggttttac caacctggat    120

ctcgctatga tcgagctgga ggactttgtc catcggtgcc cgctcccaga agacaatctt    180

gctggtcaga aggagtgaga cgatggatcc gtctagcaat gagaatgtct atgtgggtcg    240

cggtcacaac atcgaaaatg atgatgacac tgaccccagg cgttggaaga aggcgaatat    300

cagttccaac accatctccg atattcagat gacgaatggc gaagacgtac aatcagggag    360

ccctacccga acggaagttg taagcccacg tctggattat ggatcggtcg actcctcctc    420

cagcctttat tctggcagcg agcacggaaa tcaagctgag attcaaaaag agctgtccgt    480

cttgttctcg aacatgtctt tgccaggcaa cgatcggcgc ccggacgaat acattctcgt    540

gcatcaaacg ggacaagatg cttttactgg tattgccaaa ggcaacctcg accaaatgcc    600

caccaaggcg gaatttaacg cgtgctgccg tctctacagg gacggagccg gtaattacta    660

cccgccacct ctcgcattcg acaagattag cgttccagag caactggagg aaaaatgggg    720

gatgatggag gcgaaggaac gtaacaaact gcggtttcag tacaagttgg acgtatggaa    780

tcatgcgcac gctgatatgg ggatcacggg cacagagatc ttttatcaaa cagataagaa    840

cataaagctc gaccggaatt ataaactaag acctgaagac cgatacgtac aaacagaaaa    900

atacgggcgc cgggaaattc aaaagcgata tcaacacgaa ctccaggctg gttcgctgct    960

gcccgatatt atgatcaaaa ctccccaaaa tgacatccac ttcgtgtaca ggtttgccgg   1020

cgactaattg actaaccggt cgagccaatt cctccgataa cttcgtataa tgtatgctat   1080

acgaagttat ggtaccgcgg ccgcgtagag gatctgttga tcagcagttc aacctgttga   1140

tagtacgtac taagctctca tgtttcacgt actaagctct catgtttaac gtactaagct   1200

ctcatgttta acgaactaaa ccctcatggc taacgtacta agctctcatg gctaacgtac   1260

taagctctca tgtttcacgt actaagctct catgtttgaa caataaaatt aatataaatc   1320

agcaacttaa atagcctcta aggttttaag ttttataaga aaaaaaagaa tatataaggc   1380

ttttaaagct tttaaggttt aacggttgtg gacaacaagc cagggatgta acgcactgag   1440

aagcccttag agcctctcaa agcaattttg agtgacacag gaacacttaa cggctgacat   1500

gggaattagc ttcacgctgc cgcaagcact cagggcgcaa gggctgctaa aggaagcgga   1560

acacgtagaa agccagtccg cagaaacggt gctgaccccg gatgaatgtc agctactggg   1620

ctatctggac aagggaaaac gcaagcgcaa agagaaagca ggtagcttgc agtgggctta   1680

catggcgata gctagactgg gcggttttat ggacagcaag cgaaccggaa ttgccagctg   1740

gggcgccctc tggtaaggtt gggaagccct gcaaagtaaa ctggatggct ttcttgccgc   1800

caaggatctg atggcgcagg ggatcaagat ctgatcaaga gacaggatga ggatcgtttc   1860

gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat   1920

tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt   1980

cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac   2040

tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg   2100

tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc   2160

aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa   2220

tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc   2280

gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg   2340

aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg   2400

acggcgagga tctcgtcgtg acacatggcg atgcctgctt gccgaatatc atggtggaaa   2460

atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg   2520

acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct   2580

tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc   2640

ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa   2700

cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat   2760

cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt   2820

cgcccacccc gggatatccg gatatagttc ctcctttcag caaaaaaccc ctcaagaccc   2880

gtttagaggc cccaaggggt tatgctagtt attgctcagc ggtggcagca gccaactcag   2940

cttcctttcg ggctttgtta gcagccggat cttctagaat ccccagcatg cctgctattg   3000

tcttcccaat cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac   3060

ctactcagac aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca   3120

ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa   3180

ggcacagtcg aggctgatag cgagcttcaa tggtgatggt gatgatggct agaatcgaga   3240

ccgaggagag ggttagggat aggcttaccg agctcccgaa ttgccctttg aatgaatgat   3300

gatgacactg accccaggcg ttggaagaag gcgaatatca gttccaacac catctccgat   3360

attcagatga cgaatggcga agacgtacaa tcagggagcc ctacccgaac ggaagttgta   3420

agcccacgtc tggattatgg atcggtcgac tcctcctcca gcctttattc tggcagcgag   3480

cacggaaatc aagctgagat tcaaaaagag ctgtccgtct tgttctcgaa catgtctttg   3540

ccaggcaacg atcggcgccc ggacgaatac attctcgtgc atcaaacggg acaagatgct   3600

tttactggta ttgccaaagg caacctcgac caaatgccca ccaaggcgga atttaacgcg   3660

tgctgccgtc tctacaggga cggagccggt aattactacc cgccacctct cgcattcgac   3720

aagattagcg ttccagagca actggaggaa aaatggggga tgatggaggc gaaggaacgt   3780

aacaaactgc ggtttcagta caagttggac gtatggaatc atgcgcacgc tgatatgggg   3840

atcacgggca cagagatctt ttatcaaaca gataagaaca taaagctcga ccggaattat   3900

aaactaagac ctgaagaccg atacgtacaa acagaaaaat acgggcgccg ggaaattcaa   3960

aagcgatatc aacacgaact ccaggctggt tcgctgctgc ccgatattat gatcaaaact   4020

ccccaaaatg acatccactt cgtgtacagg tttgccggcg acaattacgc caacaaacag   4080

ttcagcgagt ttgaacacac cgtcaagcgc aggtatggcg acgagactga gatcaaattg   4140

aagtcaaagt caggcattat gcatgactcg aaatatctgg aatcctggga acggggcagt   4200

gcggatattc gcttcgcgga attcgttggg gaaaatagag ctcacaatcg gcagtttcca   4260

actgcgacag taaatatggg acagcagcca gacgggcagg gcggtttgac ccgcgaccgt   4320

catgtgagcg ttgacttcct aatgcaaagc gcacccaatt cgccttgggc gcaagctttg   4380

aaaaagggag aactgtggga tcgcgttcag ttgcttgctc gcgacggcaa ccgctatctg   4440

tcgccgccca gattggaata ttctgaccct gcacatttca ccgagttgat gaaccgggtt   4500

ggtttacccg catcgatggg tcggcaaagc catgcggcta gtatcaaatt cgaaaagttt   4560

gacgcgcagg cagcggttat tgtcttaaat ggcccagagt tacgtgacat tcatgacttg   4620

tctcctgaaa aactgcaaaa tttgtccacc aaagatgtca tcgtcgccga tcgcaatgag   4680

aatggtcaga gaactggcac gtacaccagc gtcgcggaat atgagcgctt gcagttaagg   4740

ctgccacccg atgcagcggg ggtgcttggt gaagcaactg acaaatattc acgtgatttc   4800

gttcggccag agccggcgtc gcgtccaatc agtgacagcc gcaggatata cgaaagtcga   4860

ccgcgtagcc aaagcgtcaa cagcttttga cgttcctgct gccgcgtcaa cgaggaagct   4920

cgtttgaccc gggtttgcca atgaaagggc tcaatcatgg tgaacactac aaagaaaagt   4980

tttgcgaagt cgcttacggc agatatgcgc cgttctgctc agcgcgttgt cgagcaaatg   5040

ygaaamgcat tgattaccga agaagaggcg ctcaagcggc aagccagact ggagagtccc   5100

gataggaagc gaaagtatgc tgctgatatg gcgatagtcg acaaactcga cgtagggttt   5160

cgaggcgaaa taggctataa aattcttgga aataaccggc ttcgagtaga caaccataaa   5220

gaattaacgc gtgagcacgg tagacttcgc aaaaccaaaa cggttctgaa gcgtaacccg   5280

gtgacgcagg aagtctattt gggtttatat gaaaggaagt cctggttaag tgtcagcagc   5340

catttgtatg ctgcggacgg cacactccgc atgaagcacg tgaaatacaa agacggacgt   5400

tttgaggaaa aatgggagcg cgacgaaaat ggcgacctga tccgcacaag gtacgccaac   5460

cgtggcaggc tctttcaacc tgtatccgag aaaatgggcg cgccgtatcg gagcggccct   5520

gacgaccggc tctatcgcga tctaacccgt cgaaacggtt tcagacggga gacattcgaa   5580

cgggacgatc acggaaacct cgagcgtatc ggcagcaacc atgtcggctt ttccaagatt   5640

tcagtgaagg cagccaatcg tc                                            5662


<210>  4
<211>  267
<212>  PRT
<213>  Agrobacterium tumefaciens

<400>  4

Met Ile Val His Pro Ser Arg Glu Asn Phe Ser Ser Ala Val Asn Lys 
1               5                   10                  15      


Gly Ser Asp Phe Arg Leu Lys Gly Glu Pro Leu Lys His Val Leu Leu 
            20                  25                  30          


Ile Asp Asp Asp Val Ala Met Arg His Leu Ile Ile Glu Tyr Leu Thr 
        35                  40                  45              


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


Arg Val Leu Ser Ser Ala Thr Val Asp Val Val Val Val Asp Leu Asn 
65                  70                  75                  80  


Leu Gly Arg Glu Asp Gly Leu Glu Ile Val Arg Asn Leu Ala Ala Lys 
                85                  90                  95      


Ser Asp Ile Pro Ile Ile Ile Ile Ser Gly Asp Arg Leu Glu Glu Thr 
            100                 105                 110         


Asp Lys Val Val Ala Leu Glu Leu Gly Ala Ser Asp Phe Ile Ala Lys 
        115                 120                 125             


Pro Phe Ser Thr Arg Glu Phe Leu Ala Arg Ile Arg Val Ala Leu Arg 
    130                 135                 140                 


Val Arg Pro Asn Val Val Arg Ser Lys Asp Arg Arg Ser Phe Cys Phe 
145                 150                 155                 160 


Thr Asp Trp Thr Leu Asn Leu Arg Gln Arg Arg Leu Met Ser Glu Ala 
                165                 170                 175     


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


Phe Leu Glu Lys Pro Arg Asp Val Leu Ser Arg Glu Gln Leu Leu Ile 
        195                 200                 205             


Ala Ser Arg Val Arg Asp Glu Glu Val Tyr Asp Arg Ser Ile Asp Val 
    210                 215                 220                 


Leu Ile Leu Arg Leu Arg Arg Lys Leu Glu Ala Asp Pro Ser Ser Pro 
225                 230                 235                 240 


Gln Leu Ile Lys Thr Ala Arg Gly Ala Gly Tyr Phe Phe Asp Ala Asp 
                245                 250                 255     


Val Gln Val Ser His Gly Gly Thr Met Ala Ala 
            260                 265         


<210>  5
<211>  65
<212>  PRT
<213>  Agrobacterium tumefaciens

<400>  5

Met Ala Ile Ile Lys Pro His Val Asn Lys Asn Arg Thr Thr Ser Pro 
1               5                   10                  15      


Ile Glu Arg Pro Glu Ser Leu Ile Glu Glu Met Ser Gly Ser His Pro 
            20                  25                  30          


Pro Ser Gly Phe Thr Asn Leu Asp Leu Ala Met Ile Glu Leu Glu Asp 
        35                  40                  45              


Phe Val His Arg Cys Pro Leu Pro Glu Asp Asn Leu Ala Gly Gln Lys 
    50                  55                  60                  


Glu 
65  


