                         SEQUENCE LISTING

<110>  TOTAL RAFFINAGE CHIMIE
 
<120>  TARGETING NUCLEAR-ENCODED RECOMBINANT PROTEINS TO THE CHLOROPLAST
        IN MICROALGAE

<130>  TOTAL-231-PCT

<150> EP16290225.8
<151> 2016-12-05

<160>  18    

<170>  PatentIn version 3.5

<210>  1
<211>  2638
<212>  DNA
<213>  Nannochloropsis gaditana

<400>  1
gttgcgtaac acttcttctg atgaggtgca ttggatggtg tcgtacgtgt tttcgcctcc       60

tcgtccacac catccgactg tttgtaccca tgatgacgcg atcacaccgt cttcacgggt      120

tgggacactg tcgtcttgct gtttgtcaac gtgccacgct acctcgcctt cgacaaagat      180

ctccatgcca tccgcggccg ggaatacctg gattttggca tagctcccac atcatgggga      240

aattcggcat gccacaccgc tcatttaact cctgagcgct gcaacatgtg caagccaagc      300

gccaagacgc ccgagccact tgttcagttc tcaagagggg ccgggaggat cgtcgcgggc      360

ttcgatgaaa gcgtctccgc ttcagtttaa gccccagcgt cattcatagt cacgttgttt      420

ttctcacaaa atctcatttt tccttgcaga agtatggtca agactgccgc cgtaagcctc      480

ctggccctag ccgggctcgc atctgccttc gtgcccccca ccacgaattt tcgcagcgct      540

aacagatgga cgattaaggc caaagacacg tccttcaccc gcaacctcat gatgaagctg      600

ggcgcggacg acaaggtcat tttgatcggc gtggccgcgg attccggctg tggaaagtcg      660

acgttcatgc ggcggctgac caacatcttt ggtgggagca acgtgggccc cctggggggc      720

ggtttcgaca acgggggatg ggagacgaac accctggtct cggacacgac caccgtcatc      780

tgcttggacg actaccatgc caacgaccgc tctgggcgga aagtgacggg ccgcaccgcc      840

ttggaagctg ccgagcagaa ttttgacctc atgtacgagc agctcaaggc cctgaaggag      900

ggcaaaactg tggccaagcc catctacaac cacgtgaacg ggaccttgga ccggcccgag      960

gaggtggtgc ccacccccat tgtgatcgtg gagggcttgc acccctggta cgacgcccgc     1020

gtcaaggacc tgctcgacta cactatctac ctggacatat cggacgagat caagcgcgca     1080

tggaagatcc agcgggacat ggccgagcgc ggatggacct tggagcaggt ggaggcagag     1140

attgaaaagc gtaagccgga cttcaataaa ttcgtggggc cccagaagga ggtagccgac     1200

tcggtgatcc aagtcttgcc cacagagctg accaacgacc ccgaggggaa gatcctccgc     1260

gtccggctca tccagaagga gacgggggac tacgaacccg tctacctgtt cgaccagggc     1320

tctacggtct cctggattcc ctgcggcacg aagctgacat gctcctaccc cggcatcaag     1380

ctgggctcgg gaccggaccg ctggttcaac aacgcggtga acgtggtgga gatggacggc     1440

cagtttgaca agctggaaga gcttgcctac gtggagaagc acctggggaa cacggccagc     1500

aagtacgacg gggagatcac ggcccagatg ctcaagaacg agggcccccg ggaccctgaa     1560

cggctcaggc ctcttccaga ccatcgtctc gctcaagatc cgcgaggtct acgagaagct     1620

gagcgggaag aaggtagacg cctccgtcaa ggcccccgtg gccgcgtaag ccggcggcaa     1680

aggagcgagg gctggttggc tgctgaaaac agggtaggct ttgaggtgtg gagatcgtaa     1740

cggctcccac cggacctcag cagcatccct tgaacaaacg gcgagcctca cacgccgact     1800

gctgcatgtt ttgtgtgttc tgtgcttttc gtgtccggag ccatcgcgtc gtctgcgccg     1860

ggtgccgggc ccagagagcg gggggagggc cagggaggca ttctgttgtt ttgggtggtg     1920

tttgggggat aggagactga cctgtcggcc cctttttgac gtatcgcgaa tttcgatgaa     1980

ataatcggct ccattctcca ttaaattgaa gcatgcatgg atgatgatcg aggtgggagg     2040

ggcgcctata acaccgccac ctgtgtccct gggcgagctc ttcgggttcc tttacttttc     2100

gactccgaaa acgcttttct gaaacaaagt cgccaaggtt actcgctgtt gcccataccc     2160

cttccattcg cgagtctaga actccttgca gatccctgga taagagattc aaaaacgttg     2220

cggccgagga gtcgatggaa tgcctccctc cttgaacccg cccggcctgc gcgtgctcat     2280

ggctcgtgac agacacccat gtcgacctcg cctgcgggag agaaacagag cgtccgaaaa     2340

cagcgcaaag ggagtccgag aactgcgcaa agaaagtccg agagcagcgc gcagcaaaag     2400

cgagggtcct tgtggcctga tttcatcgtg gaagatgtta cgaatcacga cttatgcgca     2460

ccacttcact tcatcgaagc gcgatcctgt ttatactttc tactgcacaa aattgtgtgt     2520

cgatcccctc cttctccccc tctcctcctc tccctcgcct cactcatcta aatggcgtgc     2580

cagcacatga taggtggcgt catagcaggg taaaattaca ctggccacgg gcacggcc       2638


<210>  2
<211>  428
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  In silico protein sequence of Nannochloropsis gaditana 
       phosphoribulokinase

<400>  2

Met Val Lys Thr Ala Ala Val Ser Leu Leu Ala Leu Ala Gly Leu Ala 
1               5                   10                  15      


Ser Ala Phe Val Pro Pro Thr Thr Asn Phe Arg Ser Ala Asn Arg Trp 
            20                  25                  30          


Thr Ile Lys Ala Lys Asp Thr Ser Phe Thr Arg Asn Leu Met Met Lys 
        35                  40                  45              


Leu Gly Ala Asp Asp Lys Val Ile Leu Ile Gly Val Ala Ala Asp Ser 
    50                  55                  60                  


Gly Cys Gly Lys Ser Thr Phe Met Arg Arg Leu Thr Asn Ile Phe Gly 
65                  70                  75                  80  


Gly Ser Asn Val Gly Pro Leu Gly Gly Gly Phe Asp Asn Gly Gly Trp 
                85                  90                  95      


Glu Thr Asn Thr Leu Val Ser Asp Thr Thr Thr Val Ile Cys Leu Asp 
            100                 105                 110         


Asp Tyr His Ala Asn Asp Arg Ser Gly Arg Lys Val Thr Gly Arg Thr 
        115                 120                 125             


Ala Leu Glu Ala Ala Glu Gln Asn Phe Asp Leu Met Tyr Glu Gln Leu 
    130                 135                 140                 


Lys Ala Leu Lys Glu Gly Lys Thr Val Ala Lys Pro Ile Tyr Asn His 
145                 150                 155                 160 


Val Asn Gly Thr Leu Asp Arg Pro Glu Glu Val Val Pro Thr Pro Ile 
                165                 170                 175     


Val Ile Val Glu Gly Leu His Pro Trp Tyr Asp Ala Arg Val Lys Asp 
            180                 185                 190         


Leu Leu Asp Tyr Thr Ile Tyr Leu Asp Ile Ser Asp Glu Ile Lys Arg 
        195                 200                 205             


Ala Trp Lys Ile Gln Arg Asp Met Ala Glu Arg Gly Trp Thr Leu Glu 
    210                 215                 220                 


Gln Val Glu Ala Glu Ile Glu Lys Arg Lys Pro Asp Phe Asn Lys Phe 
225                 230                 235                 240 


Val Gly Pro Gln Lys Glu Val Ala Asp Ser Val Ile Gln Val Leu Pro 
                245                 250                 255     


Thr Glu Leu Thr Asn Asp Pro Glu Gly Lys Ile Leu Arg Val Arg Leu 
            260                 265                 270         


Ile Gln Lys Glu Thr Gly Asp Tyr Glu Pro Val Tyr Leu Phe Asp Gln 
        275                 280                 285             


Gly Ser Thr Val Ser Trp Ile Pro Cys Gly Thr Lys Leu Thr Cys Ser 
    290                 295                 300                 


Tyr Pro Gly Ile Lys Leu Gly Ser Gly Pro Asp Arg Trp Phe Asn Asn 
305                 310                 315                 320 


Ala Val Asn Val Val Glu Met Asp Gly Gln Phe Asp Lys Leu Glu Glu 
                325                 330                 335     


Leu Ala Tyr Val Glu Lys His Leu Gly Asn Thr Ala Ser Lys Tyr Asp 
            340                 345                 350         


Gly Glu Ile Thr Ala Gln Met Leu Lys Asn Glu Gly Pro Arg Asp Pro 
        355                 360                 365             


Glu Arg Leu Arg Pro Leu Pro Asp His Arg Leu Ala Gln Asp Pro Arg 
    370                 375                 380                 


Gly Leu Arg Glu Ala Glu Arg Glu Glu Gly Arg Arg Leu Arg Gln Gly 
385                 390                 395                 400 


Pro Arg Gly Arg Val Ser Arg Arg Gln Arg Ser Glu Gly Trp Leu Ala 
                405                 410                 415     


Ala Glu Asn Arg Val Gly Phe Glu Val Trp Arg Ser 
            420                 425             


<210>  3
<211>  395
<212>  PRT
<213>  Arabidopsis thaliana

<400>  3

Met Ala Val Ser Thr Ile Tyr Ser Thr Gln Ala Leu Asn Ser Thr His 
1               5                   10                  15      


Phe Leu Thr Ser Ser Ser Ser Ser Lys Gln Val Phe Leu Tyr Arg Arg 
            20                  25                  30          


Gln Pro Gln Thr Asn Arg Arg Phe Asn Thr Leu Ile Thr Cys Ala Gln 
        35                  40                  45              


Glu Thr Ile Val Ile Gly Leu Ala Ala Asp Ser Gly Cys Gly Lys Ser 
    50                  55                  60                  


Thr Phe Met Arg Arg Leu Thr Ser Val Phe Gly Gly Ala Ala Lys Pro 
65                  70                  75                  80  


Pro Lys Gly Gly Asn Pro Asp Ser Asn Thr Leu Ile Ser Asp Thr Thr 
                85                  90                  95      


Thr Val Ile Cys Leu Asp Asp Tyr His Ser Leu Asp Arg Tyr Gly Arg 
            100                 105                 110         


Lys Glu Gln Lys Val Thr Ala Leu Asp Pro Arg Ala Asn Asp Phe Asp 
        115                 120                 125             


Leu Met Tyr Glu Gln Val Lys Ala Leu Lys Asn Gly Ile Ala Val Glu 
    130                 135                 140                 


Lys Pro Ile Tyr Asn His Val Thr Gly Leu Leu Asp Pro Pro Glu Leu 
145                 150                 155                 160 


Ile Gln Pro Pro Lys Ile Leu Val Ile Glu Gly Leu His Pro Met Phe 
                165                 170                 175     


Asp Glu Arg Val Arg Asp Leu Leu Asp Phe Ser Ile Tyr Leu Asp Ile 
            180                 185                 190         


Ser Asn Glu Val Lys Phe Ala Trp Lys Ile Gln Arg Asp Met Ala Glu 
        195                 200                 205             


Arg Gly His Ser Leu Glu Ser Ile Lys Ala Ser Ile Glu Ala Arg Lys 
    210                 215                 220                 


Pro Asp Phe Asp Ala Phe Ile Asp Pro Gln Lys Gln Tyr Ala Asp Ala 
225                 230                 235                 240 


Val Ile Glu Val Leu Pro Thr Thr Leu Ile Pro Asp Asp Asn Glu Gly 
                245                 250                 255     


Lys Val Leu Arg Val Arg Leu Ile Met Lys Glu Gly Val Lys Tyr Phe 
            260                 265                 270         


Ser Pro Val Tyr Leu Phe Asp Glu Gly Ser Thr Ile Ser Trp Ile Pro 
        275                 280                 285             


Cys Gly Arg Lys Leu Thr Cys Ser Tyr Pro Gly Ile Lys Phe Asn Tyr 
    290                 295                 300                 


Glu Pro Asp Ser Tyr Phe Asp His Glu Val Ser Val Leu Glu Met Asp 
305                 310                 315                 320 


Gly Gln Phe Asp Arg Leu Asp Glu Leu Ile Tyr Val Glu Ser His Leu 
                325                 330                 335     


Ser Asn Leu Ser Thr Lys Phe Tyr Gly Glu Val Thr Gln Gln Met Leu 
            340                 345                 350         


Lys His Ala Asp Phe Pro Gly Ser Asn Asn Gly Thr Gly Leu Phe Gln 
        355                 360                 365             


Thr Ile Val Gly Leu Lys Ile Arg Asp Leu Tyr Glu Gln Leu Ile Ala 
    370                 375                 380                 


Asn Lys Ala Thr Ala Arg Ala Glu Ala Lys Ala 
385                 390                 395 


<210>  4
<211>  402
<212>  PRT
<213>  Spinacia oleracea

<400>  4

Met Ala Val Cys Thr Val Tyr Thr Ile Pro Thr Thr Thr His Leu Gly 
1               5                   10                  15      


Ser Ser Phe Asn Gln Asn Asn Lys Gln Val Phe Phe Asn Tyr Lys Arg 
            20                  25                  30          


Ser Ser Ser Ser Asn Asn Thr Leu Phe Thr Thr Arg Pro Ser Tyr Val 
        35                  40                  45              


Ile Thr Cys Ser Gln Gln Gln Thr Ile Val Ile Gly Leu Ala Ala Asp 
    50                  55                  60                  


Ser Gly Cys Gly Lys Ser Thr Phe Met Arg Arg Leu Thr Ser Val Phe 
65                  70                  75                  80  


Gly Gly Ala Ala Glu Pro Pro Lys Gly Gly Asn Pro Asp Ser Asn Thr 
                85                  90                  95      


Leu Ile Ser Asp Thr Thr Thr Val Ile Cys Leu Asp Asp Phe His Ser 
            100                 105                 110         


Leu Asp Arg Asn Gly Arg Lys Val Glu Lys Val Thr Ala Leu Asp Pro 
        115                 120                 125             


Lys Ala Asn Asp Phe Asp Leu Met Tyr Glu Gln Val Lys Ala Leu Lys 
    130                 135                 140                 


Glu Gly Lys Ala Val Asp Lys Pro Ile Tyr Asn His Val Ser Gly Leu 
145                 150                 155                 160 


Leu Asp Pro Pro Glu Leu Ile Gln Pro Pro Lys Ile Leu Val Ile Glu 
                165                 170                 175     


Gly Leu His Pro Met Tyr Asp Ala Arg Val Arg Glu Leu Leu Asp Phe 
            180                 185                 190         


Ser Ile Tyr Leu Asp Ile Ser Asn Glu Val Lys Phe Ala Trp Lys Ile 
        195                 200                 205             


Gln Arg Asp Met Lys Glu Arg Gly His Ser Leu Glu Ser Ile Lys Ala 
    210                 215                 220                 


Ser Ile Glu Ser Arg Lys Pro Asp Phe Asp Ala Tyr Ile Asp Pro Gln 
225                 230                 235                 240 


Lys Gln His Ala Asp Val Val Ile Glu Val Leu Pro Thr Glu Leu Ile 
                245                 250                 255     


Pro Asp Asp Asp Glu Gly Lys Val Leu Arg Val Arg Met Ile Gln Lys 
            260                 265                 270         


Glu Gly Val Lys Phe Phe Asn Pro Val Tyr Leu Phe Asp Glu Gly Ser 
        275                 280                 285             


Thr Ile Ser Trp Ile Pro Cys Gly Arg Lys Leu Thr Cys Ser Tyr Pro 
    290                 295                 300                 


Gly Ile Lys Phe Ser Tyr Gly Pro Asp Thr Phe Tyr Gly Asn Glu Val 
305                 310                 315                 320 


Thr Val Val Glu Met Asp Gly Met Phe Asp Arg Leu Asp Glu Leu Ile 
                325                 330                 335     


Tyr Val Glu Ser His Leu Ser Asn Leu Ser Thr Lys Phe Tyr Gly Glu 
            340                 345                 350         


Val Thr Gln Gln Met Leu Lys His Gln Asn Phe Pro Gly Ser Asn Asn 
        355                 360                 365             


Gly Thr Gly Phe Phe Gln Thr Ile Ile Gly Leu Lys Ile Arg Asp Leu 
    370                 375                 380                 


Phe Glu Gln Leu Val Ala Ser Arg Ser Thr Ala Thr Ala Thr Ala Ala 
385                 390                 395                 400 


Lys Ala 
        


<210>  5
<211>  396
<212>  PRT
<213>  Phaeodactylum tricornutum

<400>  5

Met Lys Phe Ala Val Phe Ala Ser Leu Thr Ala Thr Ala Ala Ala Phe 
1               5                   10                  15      


Ala Pro Thr Ala Phe Val Pro Ser Asn Leu Arg Gly Val Ala Pro Ser 
            20                  25                  30          


Ala Ser Ser Leu Asn Met Ala Leu Lys Glu Gly Gln Thr Pro Ile Ile 
        35                  40                  45              


Ile Gly Val Ala Ala Asp Ser Gly Cys Gly Lys Ser Thr Phe Met Arg 
    50                  55                  60                  


Arg Leu Thr Asn Ile Phe Gly Gly Asp Val Val Gly Pro Leu Gly Gly 
65                  70                  75                  80  


Gly Phe Asp Lys Gly Ser Trp Glu Thr Asn Thr Leu Val Ser Asp Leu 
                85                  90                  95      


Thr Thr Val Ile Cys Leu Asp Asp Tyr His Leu Asn Asp Arg Ala Gly 
            100                 105                 110         


Arg Lys Val Thr Met Arg Thr Ala Leu Asp Pro Glu Glu Asn Asn Phe 
        115                 120                 125             


Asp Leu Met Tyr Glu Gln Val Lys Ala Leu Lys Asp Gly Lys Thr Val 
    130                 135                 140                 


Glu Lys Pro Ile Tyr Asn His Val Asn Gly Thr Leu Asp Thr Pro Glu 
145                 150                 155                 160 


Thr Ile Glu Pro Thr Pro Ile Ile Ile Phe Glu Gly Leu His Pro Met 
                165                 170                 175     


His Asp Lys Arg Val Leu Asp Leu Leu Asp Phe Ser Leu Tyr Leu Asp 
            180                 185                 190         


Ile Ser Asp Asp Val Lys Leu Asn Trp Lys Val Gln Arg Asp Met Glu 
        195                 200                 205             


Glu Arg Gly His Ser Met Glu Ser Ile Leu Ala Ser Ile Glu Ala Arg 
    210                 215                 220                 


Lys Pro Asp Phe Asp Ala Tyr Ile Asp Pro Gln Lys Gln Leu Ala Asp 
225                 230                 235                 240 


Leu Ile Ile Glu Val Leu Pro Thr Arg Leu Asp Gln Asp Asp Lys Lys 
                245                 250                 255     


Thr Leu Arg Val Arg Cys Ile Gln Lys Glu Gly Val Glu Asn Phe Asp 
            260                 265                 270         


Pro Cys Phe Leu Phe Asp Glu Gly Ser Ser Ile Glu Trp Thr Pro Ala 
        275                 280                 285             


Pro Thr Lys Leu Ser Ser Pro Ala Pro Gly Ile Lys Leu Ala Tyr Tyr 
    290                 295                 300                 


Pro Glu Glu Phe Phe Gly Lys Asp Ala Gln Val Leu Glu Met Asp Gly 
305                 310                 315                 320 


Asn Phe Asp Asn Ile Gln Glu Leu Val Tyr Val Glu Ser Ala Leu Ser 
                325                 330                 335     


Asn Thr Lys Thr Lys Phe Tyr Gly Glu Met Thr Gln Ala Met Leu Ala 
            340                 345                 350         


Leu Ala Thr Ala Pro Gly Ser Asn Asn Gly Thr Gly Leu Met Gln Thr 
        355                 360                 365             


Leu Ala Ala Phe Ala Ile Arg Asp Ile Tyr Glu Lys Lys Thr Ala Ala 
    370                 375                 380                 


Ala Lys Ala Lys Ala Gly Val Ser Ala Ala Ala Ala 
385                 390                 395     


<210>  6
<211>  375
<212>  PRT
<213>  Chlamydomonas reinhardtii

<400>  6

Met Ala Phe Thr Met Arg Ala Pro Ala Pro Arg Ala Thr Ala Gln Ser 
1               5                   10                  15      


Arg Val Thr Ala Asn Arg Ala Arg Arg Ser Leu Val Val Arg Ala Asp 
            20                  25                  30          


Lys Asp Lys Thr Val Val Ile Gly Leu Ala Ala Asp Ser Gly Cys Gly 
        35                  40                  45              


Lys Ser Thr Phe Met Arg Arg Met Thr Ser Ile Phe Gly Gly Val Pro 
    50                  55                  60                  


Lys Pro Pro Ala Gly Gly Asn Pro Asp Ser Asn Thr Leu Ile Ser Asp 
65                  70                  75                  80  


Met Thr Thr Val Ile Cys Leu Asp Asp Tyr His Cys Leu Asp Arg Asn 
                85                  90                  95      


Gly Arg Lys Val Lys Gly Val Thr Ala Leu Ala Pro Glu Ala Gln Asn 
            100                 105                 110         


Phe Asp Leu Met Tyr Asn Gln Val Lys Ala Leu Lys Glu Gly Lys Ser 
        115                 120                 125             


Val Asp Lys Pro Ile Tyr Asn His Val Ser Gly Leu Ile Asp Ala Pro 
    130                 135                 140                 


Glu Lys Ile Glu Ser Pro Pro Ile Leu Val Ile Glu Gly Leu His Pro 
145                 150                 155                 160 


Phe Tyr Asp Lys Arg Val Ala Glu Leu Leu Asp Phe Lys Ile Tyr Leu 
                165                 170                 175     


Asp Ile Ser Asp Asp Ile Lys Phe Ala Trp Lys Ile Gln Arg Asp Met 
            180                 185                 190         


Ala Glu Arg Gly His Ser Leu Glu Ser Ile Lys Ser Ser Ile Ala Ala 
        195                 200                 205             


Arg Lys Pro Asp Phe Asp Ala Tyr Ile Asp Pro Gln Lys Lys Asp Ala 
    210                 215                 220                 


Asp Met Ile Ile Gln Val Leu Pro Thr Gln Leu Val Pro Asp Asp Lys 
225                 230                 235                 240 


Gly Gln Tyr Leu Arg Val Arg Leu Ile Met Lys Glu Gly Ser Lys Met 
                245                 250                 255     


Phe Asp Pro Val Tyr Leu Phe Asp Glu Gly Ser Thr Ile Ser Trp Ile 
            260                 265                 270         


Pro Cys Gly Arg Lys Leu Thr Cys Ser Phe Pro Gly Ile Lys Met Phe 
        275                 280                 285             


Tyr Gly Pro Asp Thr Trp Tyr Gly Gln Glu Val Ser Val Leu Glu Met 
    290                 295                 300                 


Asp Gly Gln Phe Asp Lys Leu Glu Glu Leu Ile Tyr Val Glu Ser His 
305                 310                 315                 320 


Leu Ser Asn Thr Ser Ala Lys Phe Tyr Gly Glu Ile Thr Gln Gln Met 
                325                 330                 335     


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


Gln Thr Ile Val Gly Leu Lys Val Arg Glu Val Tyr Glu Arg Ile Val 
        355                 360                 365             


Lys Lys Asp Val Val Pro Val 
    370                 375 


<210>  7
<211>  403
<212>  PRT
<213>  Oryza sativa

<400>  7

Met Ala Ile Ser Ser Leu His Ala Thr Thr Ser Leu His Ser Pro Cys 
1               5                   10                  15      


Thr Thr Asn Thr Ser Phe Arg Gln Asn Gln Val Ile Phe Phe Thr Thr 
            20                  25                  30          


Arg Ser Asn Arg Arg Gly Ser Thr Arg Tyr Gly Gly Ala Arg Thr Phe 
        35                  40                  45              


Gln Val Ser Cys Ser Val Asp Lys Pro Val Val Ile Gly Leu Ala Ala 
    50                  55                  60                  


Asp Ser Gly Cys Gly Lys Ser Thr Phe Met Arg Arg Leu Thr Ser Val 
65                  70                  75                  80  


Phe Gly Gly Ala Ala Glu Pro Pro Lys Gly Gly Asn Pro Asp Ser Asn 
                85                  90                  95      


Thr Leu Ile Ser Asp Thr Thr Thr Val Ile Cys Leu Asp Asp Tyr His 
            100                 105                 110         


Ser Leu Asp Arg Thr Gly Arg Lys Glu Lys Gly Val Thr Ala Leu Asp 
        115                 120                 125             


Pro Arg Ala Asn Asp Phe Asp Leu Met Tyr Glu Gln Val Lys Ala Ile 
    130                 135                 140                 


Lys Glu Gly Lys Ala Ile Glu Lys Pro Ile Tyr Asn His Val Thr Gly 
145                 150                 155                 160 


Leu Leu Asp Pro Pro Glu Leu Ile Gln Pro Pro Lys Ile Phe Val Ile 
                165                 170                 175     


Glu Gly Leu His Pro Met Phe Asp Glu Arg Val Arg Asp Leu Leu Asp 
            180                 185                 190         


Phe Ser Ile Tyr Leu Asp Ile Ser Asp Glu Val Lys Phe Ala Trp Lys 
        195                 200                 205             


Ile Gln Arg Asp Met Ala Glu Arg Gly His Ser Leu Glu Ser Ile Lys 
    210                 215                 220                 


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


Gln Lys Gln Tyr Ala Asp Ala Val Ile Glu Val Leu Pro Thr Gln Leu 
                245                 250                 255     


Ile Pro Asp Asp Asn Glu Gly Lys Val Leu Arg Val Lys Leu Ile Met 
            260                 265                 270         


Lys Glu Gly Val Lys Asn Phe Asn Pro Val Tyr Leu Phe Asp Glu Gly 
        275                 280                 285             


Ser Ser Ile Thr Trp Val Pro Cys Gly Arg Lys Leu Thr Cys Ser Tyr 
    290                 295                 300                 


Pro Gly Ile Lys Phe Ala Tyr Gly Pro Asp Thr Tyr Phe Gly His Glu 
305                 310                 315                 320 


Val Ser Val Leu Glu Met Asp Gly Gln Phe Asp Arg Leu Asp Glu Leu 
                325                 330                 335     


Ile Tyr Val Glu Ser His Leu Ser Asn Leu Ser Thr Lys Phe Tyr Gly 
            340                 345                 350         


Glu Val Thr Gln Gln Met Leu Lys His Ala Asp Phe Pro Gly Ser Asn 
        355                 360                 365             


Asn Gly Thr Gly Leu Phe Gln Thr Ile Val Gly Leu Lys Ile Arg Asp 
    370                 375                 380                 


Leu Tyr Glu Gln Ile Ile Ala Glu Arg Ala Gly Ala Pro Thr Glu Ala 
385                 390                 395                 400 


Ala Lys Val 
            


<210>  8
<211>  52
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  bipartite targeting sequence of the phoshoribulokinase of 
       Nannochloropsis gaditana (NgPRK BTS)

<400>  8

Met Val Lys Thr Ala Ala Val Ser Leu Leu Ala Leu Ala Gly Leu Ala 
1               5                   10                  15      


Ser Ala Phe Val Pro Pro Thr Thr Asn Phe Arg Ser Ala Asn Arg Trp 
            20                  25                  30          


Thr Ile Lys Ala Lys Asp Thr Ser Phe Thr Arg Asn Leu Met Met Lys 
        35                  40                  45              


Leu Gly Ala Asp 
    50          


<210>  9
<211>  156
<212>  DNA
<213>  Nannochloropsis gaditana

<400>  9
atggtcaaga ctgccgccgt aagcctcctg gccctagccg ggctcgcatc tgccttcgtg       60

ccccccacca cgaattttcg cagcgctaac agatggacga ttaaggccaa agacacgtcc      120

ttcacccgca acctcatgat gaagctgggc gcggac                                156


<210>  10
<211>  720
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  codon-optimized (for expression in Nannochloropsis) eYFP

<400>  10
atggtctcca agggcgagga gctcttcacc ggcgtcgtcc ccatcctcgt cgagctcgac       60

ggcgacgtca acggccacaa gttctccgtc tccggcgagg gcgagggcga cgctacctac      120

ggcaagctca ccctcaagtt catctgcacc accggcaagc tccccgtccc ctggcccacc      180

ctcgtcacca ccttcggcta cggcctccag tgcttcgctc gctaccccga ccacatgaag      240

cagcacgact tcttcaagtc cgctatgccc gagggctacg tccaggagcg caccatcttc      300

ttcaaggacg acggcaacta caagacccgc gctgaggtca agttcgaggg cgacaccctc      360

gtcaaccgca tcgagctcaa gggcatcgac ttcaaggagg acggcaacat cctcggccac      420

aagctcgagt acaactacaa ctcccacaac gtctacatca tggctgacaa gcagaagaac      480

ggcatcaagg tcaacttcaa gatccgccac aacatcgagg acggctccgt ccagctcgct      540

gaccactacc agcagaacac ccccatcggc gacggccccg tcctcctccc cgacaaccac      600

tacctctcct accagtccgc tctctccaag gaccccaacg agaagcgcga ccacatggtc      660

ctcctcgagt tcgtcaccgc tgctggcatc accctcggca tggacgagct ctacaagtaa      720


<210>  11
<211>  31
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer eYFP Fw2

<400>  11
ccgccggaat tcatggtctc caagggcgag g                                      31


<210>  12
<211>  35
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer eYFP Rev2

<400>  12
gaaagtccat atgttacttg tagagctcgt ccatg                                  35


<210>  13
<211>  5198
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  pCT2Ng vector

<400>  13
gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt       60

caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa      120

ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt      180

gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt      240

tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt      300

ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg      360

tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga      420

atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa      480

gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga      540

caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa      600

ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca      660

ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta      720

ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac      780

ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc      840

gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag      900

ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga      960

taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt     1020

agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata     1080

atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag     1140

aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa     1200

caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt     1260

ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc     1320

cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa     1380

tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa     1440

gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc     1500

ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa     1560

gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa     1620

caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg     1680

ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc     1740

tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg     1800

ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg     1860

agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg     1920

aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat     1980

gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg     2040

tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt     2100

tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg     2160

ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctcagc tgctgccccg     2220

accgtatctc caagtcagac atgaaatctt cagttgcgtt aaaaactcta cgatgctacc     2280

agcgttaaat aaccttgccc acgcctttaa acgtacccga tcattaacat atcgactggc     2340

tgccttggct ttgcaccagc catcatcaga cttaacgatg ggtatgttgc ttgcctttcc     2400

tgcttgaagg gggtccgact ctctgctttc tcgatcgcgg gtgtgacctc tgaattggaa     2460

tgtaaaaatg taagaagcga cgtgtccggt aaagaaatgc ccaagctcca tcaaatctgc     2520

gttgtcggcg accaaaccat gctggctcgt cgacctgccc cggatgcagg agcatggcac     2580

tcggcggcat ggcacttgag cctcgcggga ggaatgtgtg tggttgggcg caggctgtgg     2640

acggcccccc tccagcgaag cggtcgcctc cctttccgac gctttgtgca cgttgtctgg     2700

tgtcctctgt ctcacgcacc tcttcaccga cgtggtgtcc ctcttgttgc tggtgaggga     2760

cttggaatgt ggtcctggtt ctatcctggg cgcgtgtgtt cctttttttc tctaccgtta     2820

ttctctccat ttctgatgtc tcaccaccat ctccctcacc ctccaaccgc gtcgttgtgc     2880

caaaatcata cagcaggatc gatggccaag ttgaccagtg ccgttccggt gctcaccgcg     2940

cgcgacgtcg ccggagcggt cgagttctgg accgaccggc tcgggttctc ccgggacttc     3000

gtggaggacg acttcgccgg tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc     3060

caggaccagg tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg cctggacgag     3120

ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc     3180

atgaccgaga tcggcgagca gccgtggggg cgggagttcg ccctgcgcga cccggccggc     3240

aactgcgtgc acttcgtggc cgaggagcag gactaaatcg atcttcctta aaaatttaat     3300

tttcattagt tgcagtcact ccgctttggt ttcacagtca ggaataacac tagctcgtct     3360

tcaccatgga tgccaatctc gcctattcat ggtgtataaa agttcaacat ccaaagctag     3420

aacttttgga aagagaaaga atatccgaat agggcacggc gtgccgtatt gttggagtgg     3480

actagcagaa agtgaggaag gcacaggatg agttttctcg agagctgctg ccccgaccgt     3540

atctccaagt cagacatgaa atcttcagtt gcgttaaaaa ctctacgatg ctaccagcgt     3600

taaataacct tgcccacgcc tttaaacgta cccgatcatt aacatatcga ctggctgcct     3660

tggctttgca ccagccatca tcagacttaa cgatgggtat gttgcttgcc tttcctgctt     3720

gaagggggtc cgactctctg ctttctcgat cgcgggtgtg acctctgaat tggaatgtaa     3780

aaatgtaaga agcgacgtgt ccggtaaaga aatgcccaag ctccatcaaa tctgcgttgt     3840

cggcgaccaa accatgctgg ctcgtcgacc tgccccggat gcaggagcat ggcactcggc     3900

ggcatggcac ttgagcctcg cgggaggaat gtgtgtggtt gggcgcaggc tgtggacggc     3960

ccccctccag cgaagcggtc gcctcccttt ccgacgcttt gtgcacgttg tctggtgtcc     4020

tctgtctcac gcacctcttc accgacgtgg tgtccctctt gttgctggtg agggacttgg     4080

aatgtggtcc tggttctatc ctgggcgcgt gtgttccttt ttttctctac cgttattctc     4140

tccatttctg atgtctcacc accatctccc tcaccctcca accgcgtcgt tgtgccaaaa     4200

tcatacagca ggaggcctgt cgacggcgcg ccggatccag atctgaattc gatatcacgc     4260

gtccatggca tatggctagc gcggccgcct cgagtctaga cttccttaaa aatttaattt     4320

tcattagttg cagtcactcc gctttggttt cacagtcagg aataacacta gctcgtcttc     4380

accatggatg ccaatctcgc ctattcatgg tgtataaaag ttcaacatcc aaagctagaa     4440

cttttggaaa gagaaagaat atccgaatag ggcacggcgt gccgtattgt tggagtggac     4500

tagcagaaag tgaggaaggc acaggatgag ttttctcgag ggtacccaat tcgccctata     4560

gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc     4620

ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata     4680

gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg     4740

acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg     4800

ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca     4860

cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta     4920

gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc     4980

catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg     5040

gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat     5100

aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta     5160

acgcgaattt taacaaaata ttaacgctta caatttag                             5198


<210>  14
<211>  891
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  pCT55 cassette

<400>  14
gaattcatga gatccttttg catcgcagcc cttttggctg tggcatctgc cttcaccaca       60

cagccaactt ccttcactgt gaagactgcg aatgtgggcg aacgggcgag tggggttttc      120

cctgagcaga gctctgctca tcgcacgcgt aaagcaacga ttgtcatggt ctccaagggc      180

gaggagctct tcaccggcgt cgtccccatc ctcgtcgagc tcgacggcga cgtcaacggc      240

cacaagttct ccgtctccgg cgagggcgag ggcgacgcta cctacggcaa gctcaccctc      300

aagttcatct gcaccaccgg caagctcccc gtcccctggc ccaccctcgt caccaccttc      360

ggctacggcc tccagtgctt cgctcgctac cccgaccaca tgaagcagca cgacttcttc      420

aagtccgcta tgcccgaggg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc      480

aactacaaga cccgcgctga ggtcaagttc gagggcgaca ccctcgtcaa ccgcatcgag      540

ctcaagggca tcgacttcaa ggaggacggc aacatcctcg gccacaagct cgagtacaac      600

tacaactccc acaacgtcta catcatggct gacaagcaga agaacggcat caaggtcaac      660

ttcaagatcc gccacaacat cgaggacggc tccgtccagc tcgctgacca ctaccagcag      720

aacaccccca tcggcgacgg ccccgtcctc ctccccgaca accactacct ctcctaccag      780

tccgctctct ccaaggaccc caacgagaag cgcgaccaca tggtcctcct cgagttcgtc      840

accgctgctg gcatcaccct cggcatggac gagctctaca agtaacatat g               891


<210>  15
<211>  885
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  pCT56 cassette

<400>  15
gaattcatgg tcaagactgc cgccgtaagc ctcctggccc tagccgggct cgcatctgcc       60

ttcgtgcccc ccaccacgaa ttttcgcagc gctaacagat ggacgattaa ggccaaagac      120

acgtccttca cccgcaacct catgatgaag ctgggcgcgg acgtctccaa gggcgaggag      180

ctcttcaccg gcgtcgtccc catcctcgtc gagctcgacg gcgacgtcaa cggccacaag      240

ttctccgtct ccggcgaggg cgagggcgac gctacctacg gcaagctcac cctcaagttc      300

atctgcacca ccggcaagct ccccgtcccc tggcccaccc tcgtcaccac cttcggctac      360

ggcctccagt gcttcgctcg ctaccccgac cacatgaagc agcacgactt cttcaagtcc      420

gctatgcccg agggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac      480

aagacccgcg ctgaggtcaa gttcgagggc gacaccctcg tcaaccgcat cgagctcaag      540

ggcatcgact tcaaggagga cggcaacatc ctcggccaca agctcgagta caactacaac      600

tcccacaacg tctacatcat ggctgacaag cagaagaacg gcatcaaggt caacttcaag      660

atccgccaca acatcgagga cggctccgtc cagctcgctg accactacca gcagaacacc      720

cccatcggcg acggccccgt cctcctcccc gacaaccact acctctccta ccagtccgct      780

ctctccaagg accccaacga gaagcgcgac cacatggtcc tcctcgagtt cgtcaccgct      840

gctggcatca ccctcggcat ggacgagctc tacaagtaac atatg                      885


<210>  16
<211>  732
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  pCT59 cassette

<400>  16
gaattcatgg tctccaaggg cgaggagctc ttcaccggcg tcgtccccat cctcgtcgag       60

ctcgacggcg acgtcaacgg ccacaagttc tccgtctccg gcgagggcga gggcgacgct      120

acctacggca agctcaccct caagttcatc tgcaccaccg gcaagctccc cgtcccctgg      180

cccaccctcg tcaccacctt cggctacggc ctccagtgct tcgctcgcta ccccgaccac      240

atgaagcagc acgacttctt caagtccgct atgcccgagg gctacgtcca ggagcgcacc      300

atcttcttca aggacgacgg caactacaag acccgcgctg aggtcaagtt cgagggcgac      360

accctcgtca accgcatcga gctcaagggc atcgacttca aggaggacgg caacatcctc      420

ggccacaagc tcgagtacaa ctacaactcc cacaacgtct acatcatggc tgacaagcag      480

aagaacggca tcaaggtcaa cttcaagatc cgccacaaca tcgaggacgg ctccgtccag      540

ctcgctgacc actaccagca gaacaccccc atcggcgacg gccccgtcct cctccccgac      600

aaccactacc tctcctacca gtccgctctc tccaaggacc ccaacgagaa gcgcgaccac      660

atggtcctcc tcgagttcgt caccgctgct ggcatcaccc tcggcatgga cgagctctac      720

aagtaacata tg                                                          732


<210>  17
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer Cass02Ng Fw

<400>  17
cttggaatgt ggtcctggtt                                                   20


<210>  18
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Primer eYFP Rev

<400>  18
gaacttgagg gtgagcttgc                                                   20


