                         SEQUENCE LISTING

<110>  SYNTHETIC GENOMICS, INC.
       VERRUTO, John
       MOELLERING, Eric
       AJJAWI, Imad
 
<120>  ALGAL LIPID PRODUCTIVITY VIA GENETIC MODIFICATION OF A TPR DOMAIN
        CONTAINING PROTEIN

<130>  SGI2130-1WO

<150>  US 62/596,671
<151>  2017-12-08

<160>  25    

<170>  PatentIn version 3.5

<210>  1
<211>  51
<212>  PRT
<213>  Nannochloropsis gaditana


<220>
<221>  misc_feature
<223>  TPR domain of polypeptide of SEQ ID NO:3 (TRP-6029)

<400>  1

Lys Ala Glu Gly Asn Glu Arg Phe Arg Asn Gly Arg Leu Arg Ser Ala 
1               5                   10                  15      


Leu Glu Cys Tyr Glu Glu Ala Val Arg Met Asp Pro Ser Glu Pro Val 
            20                  25                  30          


Tyr Tyr Ala His Arg Ser Thr Cys Leu Phe Glu Leu Gly Lys Tyr Ala 
        35                  40                  45              


Ala Ser Cys 
    50      


<210>  2
<211>  91
<212>  PRT
<213>  Nannochloropsis gaditana


<220>
<221>  misc_feature
<223>  DUF4470 of polypeptide of SEQ ID NO:3 (TRP-6029)

<400>  2

Asp Asp Gly Gly Ile Asp Leu Leu Ala Arg Pro Glu Gly Glu Arg Asp 
1               5                   10                  15      


Leu Ala Leu Leu Phe Gly Gly Leu Gly Asp Ala Arg Gln Pro Leu Ala 
            20                  25                  30          


Thr Phe Arg Asp Ile Tyr Gln Gln Val Lys Gly Ser Lys Gly Gln Leu 
        35                  40                  45              


Thr Tyr Lys Thr Met Ser Leu Ser Met Ile Leu Asn Asp Val Lys Ala 
    50                  55                  60                  


Glu Cys Leu Thr Arg Ala Val Ile Met Phe Lys Ala Leu Ala Glu Leu 
65                  70                  75                  80  


Gly Ile Ile Leu Gln Glu Ala Ala Arg Glu Gln 
                85                  90      


<210>  3
<211>  1249
<212>  PRT
<213>  Nannochloropsis gaditana


<220>
<221>  misc_feature
<223>  TRP-6029 protein

<400>  3

Met Glu Ala Arg Ser Arg Lys Leu Ala Arg Lys Asn Ala Ala Phe Glu 
1               5                   10                  15      


Val Lys Ala Glu Gly Asn Glu Arg Phe Arg Asn Gly Arg Leu Arg Ser 
            20                  25                  30          


Ala Leu Glu Cys Tyr Glu Glu Ala Val Arg Met Asp Pro Ser Glu Pro 
        35                  40                  45              


Val Tyr Tyr Ala His Arg Ser Thr Cys Leu Phe Glu Leu Gly Lys Tyr 
    50                  55                  60                  


Ala Ala Ser Cys Glu Asp Ser Lys Gln Ala Ser Leu Leu Tyr Arg Asp 
65                  70                  75                  80  


Lys Gln Gln Ile Gly Met Ile Ser Arg Thr Glu Ala Asn Ala Ala Ile 
                85                  90                  95      


Ala Arg Leu Leu Arg Gln Ser Ala Arg Ala Phe Leu Cys Val Asn Thr 
            100                 105                 110         


Pro His Ser Leu Glu Ala Ala Lys Ala Leu Leu Ala Arg Thr Ile Glu 
        115                 120                 125             


Leu His Pro Asp Pro Asp Ala Glu Leu Leu Ser Met Gln Ala Gly Ala 
    130                 135                 140                 


Cys Gln Ala Val Ala Glu Ala Asp Ser Ala Met Ala Gly Glu Gly Ser 
145                 150                 155                 160 


Gly Pro Gly Tyr Met Val Gln Ser Ser Ser Lys His Gly Arg Gly Cys 
                165                 170                 175     


His Met Gly Ile Gly Gly Gly Ser Cys Asp Tyr Leu Pro Arg Tyr Arg 
            180                 185                 190         


Ser Pro Val Val Asn Gly Pro Thr Pro Tyr Ser Cys Ile Gly Trp Glu 
        195                 200                 205             


Leu Gly Leu Ser Gly Leu Ala Gly Arg Val Pro Ser Pro Ala Asp Thr 
    210                 215                 220                 


Glu Ser Pro Phe Ser Ala His Gly Gly Thr Phe Gly Glu Asp Glu Asp 
225                 230                 235                 240 


Asp Gly Gly Ile Asp Leu Leu Ala Arg Pro Glu Gly Glu Arg Asp Leu 
                245                 250                 255     


Ala Leu Leu Phe Gly Gly Leu Gly Asp Ala Arg Gln Pro Leu Ala Thr 
            260                 265                 270         


Phe Arg Asp Ile Tyr Gln Gln Val Lys Gly Ser Lys Gly Gln Leu Thr 
        275                 280                 285             


Tyr Lys Thr Met Ser Leu Ser Met Ile Leu Asn Asp Val Lys Ala Glu 
    290                 295                 300                 


Cys Leu Thr Arg Ala Val Ile Met Phe Lys Ala Leu Ala Glu Leu Gly 
305                 310                 315                 320 


Ile Ile Leu Gln Glu Ala Ala Arg Glu Gln Gly Met Glu Gly Gly Met 
                325                 330                 335     


Glu Gly Val Asp Glu Gly Val Ala Gly Thr Gly Ser Leu Asp Pro Leu 
            340                 345                 350         


Ala Leu Leu Glu Ser Ser Asp Ala Val Ala Glu Ala Val Tyr Arg Cys 
        355                 360                 365             


Tyr His Leu Tyr Leu Gly Ala Phe Leu Met Pro Asn Glu Ala Asp Trp 
    370                 375                 380                 


Leu Gln Arg Thr Leu Asn Asp Met Ser Lys Ser Asn Ser Leu Thr Gln 
385                 390                 395                 400 


Ala Ser Ser Leu Pro Gly Ser Asp His Asp Ala Arg Val Gly Ser Ser 
                405                 410                 415     


Pro Pro Leu Ala Ser Thr Pro Ser Ala Gly Pro Ser Leu Leu Asp Ala 
            420                 425                 430         


Ser Gln Ser Ser Thr Pro Phe Gln Phe Ser Ser Asn Leu Gly Phe Ser 
        435                 440                 445             


Trp Leu Lys Ile Lys Ala Lys Lys Asp Arg Glu Ala Val Lys Asn Val 
    450                 455                 460                 


Val Asp His Trp Arg Val Leu Cys Lys Thr Met Gly Ala Asp Val Met 
465                 470                 475                 480 


Ala Arg Ile Tyr Gln Ser Ser Met Gln Arg Pro Thr Glu Glu Asp Glu 
                485                 490                 495     


Gly Gly Leu Gln Thr Ala Ser His Asp Arg Gly Gly Gly Thr Gly Gly 
            500                 505                 510         


Gly Ala Arg Ala Asp Ala Met Glu Glu Ser Asn Leu Ser Leu Glu Gln 
        515                 520                 525             


Gln Trp Arg Ala Gln Val Arg Glu Ala Met Leu Glu Gln Ile Glu Ala 
    530                 535                 540                 


Met Asp Asp Glu Glu Ile Asn Gln Met Arg Glu Ala Glu Gly Ala Thr 
545                 550                 555                 560 


Pro Glu Glu Lys Arg Ala Phe Leu Arg Asp His Trp Ala Glu Asp Val 
                565                 570                 575     


Asp Pro Arg Thr Leu Asp Leu Met Arg His Leu Pro Ala Ala His Arg 
            580                 585                 590         


Glu Ile Asp Phe Phe His Thr Thr Met Leu Leu Pro Val Pro Thr Pro 
        595                 600                 605             


Glu Ala Ala Arg Gly Met Gly Leu Met Asp Lys Ser Gln Gly Arg Asn 
    610                 615                 620                 


Leu His Asn Ala Trp Arg Pro Asn Val Thr Leu Val Ser Ala Asp Ile 
625                 630                 635                 640 


Pro Met Glu Ala Met Leu Pro Gly Lys Leu Asp Lys Ala Ala Val Ala 
                645                 650                 655     


Ser Ser Ser Leu Thr Glu Leu His Phe Cys Pro Phe Lys Ala Leu Lys 
            660                 665                 670         


Leu Leu Leu Ile Gln Pro Gly Asp Glu Asn His Leu Ala Arg Gln Glu 
        675                 680                 685             


Ala Phe Arg Ser Ala Ala Leu Phe Phe Ser Glu Val Ala Leu Gly Leu 
    690                 695                 700                 


Ala Gly Phe Leu Glu Ala Gly Thr Leu Lys Met Gln Met Phe Leu Gly 
705                 710                 715                 720 


Asp Val His Asp Leu Gly Ser Thr Arg Ala Pro Asn Ser Leu Asp Arg 
                725                 730                 735     


Val Leu Ile Ser Asn Val Pro Asp Tyr Thr Thr Leu Leu Pro Ser Met 
            740                 745                 750         


Ile Lys Leu Ile Pro Leu Leu Lys Thr Ser Pro Gly Ser Ala Leu Lys 
        755                 760                 765             


His Ser Val Leu Lys Phe Asn Ala Asn Phe Gln Asp Leu Pro Glu Tyr 
    770                 775                 780                 


Ala His Ser Met Gly Leu Tyr Val Pro Asp Met Ala Cys Leu Pro Thr 
785                 790                 795                 800 


Tyr Leu Gly Val Asn His Glu Tyr Gly Gly Ile Trp Ala His Leu Ile 
                805                 810                 815     


Glu Trp Ser Arg Ala Pro Pro Leu Leu Leu Glu Gly Pro Gly Glu Glu 
            820                 825                 830         


Ser Leu His Leu Asp Met Glu Lys Asp Glu Ser Thr Pro His Arg Thr 
        835                 840                 845             


Ser Ser Pro Ala Pro Phe Thr Thr Pro Leu Pro Pro Thr Gln His Leu 
    850                 855                 860                 


Pro Ser Gly Arg Asp Val Gln Ala Trp Leu Ser Thr Val Phe Leu Ser 
865                 870                 875                 880 


Ile Ala Met Pro Leu Cys Arg Asp Cys Val Leu Thr His Thr Glu Ile 
                885                 890                 895     


Arg Pro Leu Thr Leu Gln Ala Phe Phe Glu Leu Cys His Tyr Leu Val 
            900                 905                 910         


Ala His Met Asp Phe Pro Pro His Asn Leu Ala Trp Val Ile Glu His 
        915                 920                 925             


Ala Met Ile Gly Glu Leu Thr Thr Ala Ala Val Pro Pro Asp Lys Thr 
    930                 935                 940                 


Pro Trp Leu Pro Gln Tyr Thr Trp Arg Gln Gln Gln Leu Ser Arg Ala 
945                 950                 955                 960 


Val Pro Thr Gly Ala Phe Ser Leu Glu Thr Arg Thr Leu Ala Gly Leu 
                965                 970                 975     


Trp Gln Pro Lys Leu Gly Phe Arg Leu Cys Ala Pro Val Gly His Arg 
            980                 985                 990         


Leu Pro Arg Pro Glu Asp Val Thr  Gln Leu Arg Leu Thr  Val Pro Trp 
        995                 1000                 1005             


Arg Gln  Gln Leu Pro Cys Glu  Gly Arg Glu Ser Ala  Glu Ala Val 
    1010                 1015                 1020             


Ala Val  Gln Ala Glu Ser Cys  Leu Lys Ala Thr Gly  Val Pro Leu 
    1025                 1030                 1035             


Ala Lys  Val Val Gly Ala Val  Leu Val Ser Pro Ala  Phe Leu Ala 
    1040                 1045                 1050             


Thr His  Ala Glu Ala Phe Asp  Pro Ser Lys His Tyr  Val Gln Met 
    1055                 1060                 1065             


Pro Pro  His Gly Cys Ser His  Thr Ala Cys Thr Phe  Ser Ser Ser 
    1070                 1075                 1080             


Pro Ser  Lys Thr Gly His Glu  Pro His Pro Thr Pro  Glu Asn Gln 
    1085                 1090                 1095             


Gly Leu  Arg Pro Leu Leu Leu  Ala Pro Ala Ala Gln  Lys Ser Gly 
    1100                 1105                 1110             


Asp Val  His Leu Phe Ser Cys  Val Arg Trp Asp Ser  Arg Thr Ser 
    1115                 1120                 1125             


Leu Val  Thr Leu Leu Met Pro  Lys Asp Asp Leu Arg  Gln Leu Val 
    1130                 1135                 1140             


Ala Arg  Ser Phe His Leu Cys  Leu Leu Arg Thr Asp  Ser Trp Gln 
    1145                 1150                 1155             


Pro Leu  Thr Lys Pro Phe Pro  Leu Ser Asp Asp Pro  Ala Leu Leu 
    1160                 1165                 1170             


Arg Pro  His Ile His Ala His  Met Gln Glu Met Gln  Glu Leu Arg 
    1175                 1180                 1185             


Glu Ser  Glu Leu Gly Glu Ala  Val Gly Glu Thr Thr  Glu Ser Ala 
    1190                 1195                 1200             


Met Asp  Val Glu Ile Ala Glu  Gly Gly His Ile Gly  Ala Tyr Gly 
    1205                 1210                 1215             


Asn Pro  Ala Glu Thr Gly Arg  Gly Met Val Ser Gly  Asn Ala Ser 
    1220                 1225                 1230             


Leu Val  Ala Thr Ala Glu Glu  Arg Leu Leu His Lys  Tyr Arg Thr 
    1235                 1240                 1245             


Tyr 
    


<210>  4
<211>  3750
<212>  DNA
<213>  Nannochloropsis gaditana


<220>
<221>  misc_feature
<223>  TRP-6029 open reading frame (ORF)

<400>  4
atggaagctc gatcaagaaa gctcgcccgc aaaaatgcgg cctttgaagt caaagcggaa       60

ggaaacgaac gctttcgtaa tggccgcctg cgatcagcgt tggaatgtta tgaagaggcc      120

gtgcgcatgg atccttccga gcccgtgtac tatgcccaca ggtccacctg cctctttgag      180

ctggggaagt atgcggcctc ctgcgaagac tccaagcaag cctccctcct gtaccgggac      240

aagcaacaga tcggcatgat atcccgcacg gaggccaacg ctgccatcgc ccgcttgctt      300

cgccagtccg cccgcgcttt cctctgcgtc aacaccccac acagccttga agctgccaag      360

gccctcttgg cccgaaccat cgaacttcac cccgaccccg acgcggagct tctctccatg      420

caggcaggcg cctgtcaagc cgtggcagaa gcggactcgg ccatggcagg agaaggttca      480

gggcctggat acatggtgca atcctcctcg aaacacggta ggggatgtca catggggatc      540

ggcggtggct cctgcgatta cttgccgcgg taccgatctc ccgttgtgaa cggtcccacg      600

ccctattcct gcattggatg ggagttgggc ttgtcgggat tggccggtcg tgtcccctcg      660

cccgcggata cagagtcgcc cttctcggcc cacggaggga cgttcgggga ggacgaggac      720

gacggaggca tcgacttgtt ggctcggccg gagggtgaga gggacctggc gctcctcttc      780

gggggtttgg gggacgcgcg ccagccgctg gcgacgtttc gggacattta tcagcaagtg      840

aaggggagca agggccagtt gacttacaaa acgatgagtc tgtcgatgat cttgaacgac      900

gtgaaggcgg agtgcctgac ccgggccgtg ataatgttca aggcgctggc agagctcgga      960

atcatcctgc aggaggcggc tagggagcaa gggatggagg gggggatgga gggagtggac     1020

gagggggtgg cggggacggg aagcttagat ccactggcat tgctcgagag ctcagacgca     1080

gtagcagagg ctgtgtatcg ctgctaccac ctctacttgg gcgccttcct gatgcccaac     1140

gaagccgatt ggctgcaacg gacactaaac gacatgagta agagcaactc gctcacccag     1200

gcctccagtc tccccggcag cgaccacgat gcccgcgtcg gcagcagtcc tcccctcgcg     1260

tccacgccgt ccgcgggacc gagcctcctg gatgcatccc aatcttccac gcccttccaa     1320

ttctcctcca acctcgggtt ctcctggttg aaaatcaagg ccaagaaaga tagggaggcg     1380

gtcaagaacg tggtggacca ttggcgcgtc ctgtgcaaga ccatgggagc ggacgtgatg     1440

gcgcgcatct accaatcctc catgcagagg ccgaccgagg aggatgaggg ggggttgcag     1500

acggcttcgc acgaccgagg gggagggact ggggggggcg ccagggcgga tgccatggag     1560

gagtcaaatt tgagtttgga gcaacagtgg agggcacagg tgagggaggc gatgttggaa     1620

caaatcgagg cgatggacga cgaggagatc aatcagatgc gggaagcgga aggggcgacg     1680

ccggaagaga agcgggcctt tttgcgcgac cactgggcgg aagacgtgga tccccgcacc     1740

ttggacctca tgcggcacct gcccgcggcg caccgtgaaa tcgacttttt ccacaccacc     1800

atgctcttac cggtacccac cccggaggcc gcacggggca tgggcttgat ggacaagagc     1860

caaggccgta atttgcacaa cgcctggcgg cccaatgtga ccttggtctc ggcagacatt     1920

cccatggagg ccatgttgcc gggcaagctg gacaaagcgg ccgttgcctc ctcctccttg     1980

acagagctcc atttctgccc attcaaagcc ttgaaattgt tgttgattca gcccggggac     2040

gagaaccact tggcccgtca agaagccttc cgatcagccg cgctgttctt ctcagaagta     2100

gcactgggac tggccgggtt cttggaagca gggaccctga aaatgcaaat gttccttggc     2160

gacgtgcacg atctgggcag cacgcgcgcc cccaatagct tggaccgtgt tctcatcagc     2220

aacgtccccg actacaccac gcttcttccc tccatgatca aactcatccc cctcctcaaa     2280

acgagtcctg gatccgcctt gaagcacagt gtgctgaaat ttaatgccaa ttttcaagac     2340

ttgcccgaat acgcacactc catggggttg tatgtgcccg acatggcctg cctcccgacc     2400

tatctgggtg tgaaccacga atacggaggc atatgggccc acttgatcga gtggagtcgc     2460

gcgccgccct tgctcctgga aggccccgga gaggagagtc ttcacctcga catggagaag     2520

gacgagagca cgccccaccg gacatcctcc ccagcccctt ttaccacccc gctcccgccc     2580

acacagcacc tcccctccgg gcgcgatgtg caagcctggc tctccacggt gtttctcagt     2640

atcgccatgc ccttgtgtcg ggattgcgtg ctcacacaca ccgagatccg cccgctcacc     2700

cttcaggcct ttttcgagct ctgtcattac ctggtcgcac acatggactt cccaccccac     2760

aatttggctt gggtcatcga gcacgccatg atcggagagc ttaccactgc tgcggtaccc     2820

cctgacaaga caccctggct gccccagtac acatggcgcc agcagcagct ttcgcgcgca     2880

gtgcccacgg gggctttctc cctggaaacg cgaacgctcg ccgggctatg gcagccgaaa     2940

cttggtttcc gcctgtgtgc gccagtcgga catcgcttgc cacggccaga ggacgtcacg     3000

cagctgcgct tgacggtacc gtggcgccag cagcttccct gtgaagggag agaaagtgcg     3060

gaagcggtgg ctgttcaagc cgaatcctgt ctgaaagcga cgggcgtacc gctggcaaag     3120

gtcgtgggcg ccgtcttagt ctcccctgct tttctagcga cgcatgccga agccttcgat     3180

ccctccaaac attacgtgca aatgcctcct cacggttgct ctcacaccgc ctgcactttc     3240

tcttcctcgc ccagcaagac tggccacgaa ccccatccaa ctcccgaaaa ccaagggctg     3300

cggcccctcc tcctggctcc tgcagcgcaa aaaagtggtg acgtacacct cttctcttgt     3360

gtccgttggg actcacggac ctcgctggtc acccttctca tgcccaagga cgacttgcgg     3420

cagctcgttg ccaggagttt ccatctctgt ttactgcgaa cggacagttg gcaacccctc     3480

accaagcctt tccccctctc ggacgacccg gccttgctcc ggccgcacat ccatgcgcac     3540

atgcaggaga tgcaggaatt gcgggaaagt gaactcgggg aggccgtggg agagacgact     3600

gaatccgcca tggacgtgga gatcgcggaa gggggccaca taggggccta cgggaatcca     3660

gcagagaccg gcagagggat ggtatcgggg aatgcctcgc ttgtggcaac tgctgaggag     3720

cgtttactgc acaaataccg cacatattga                                      3750


<210>  5
<211>  23
<212>  DNA
<213>  Nannochloropsis gaditana


<220>
<221>  misc_feature
<223>  Target sequence in TPR-6029 gene (PAM underlined)

<400>  5
ggatgtcaca tggggatcgg cgg                                               23


<210>  6
<211>  11263
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic


<220>
<221>  misc_feature
<223>  Vector pSGE-6206

<400>  6
gcggccgccg tatggtcgac ggttgctcgg atgggggggg cggggagcga tggagggagg       60

aagatcaggt aaggtctcga cagactagag aagcacgagt gcaggtataa gaaacagcaa      120

aaaaaagtaa tgggcccagg cctggagagg gtatttgtct tgtttttctt tggccaggaa      180

cttgttctcc tttcttcgtt tctaggaccc cgatccccgc tcgcatttct ctcttcctca      240

gccgaagcgc agcggtaaag catccatttt atcccaccga aagggcgctc ccagccttcg      300

tcgagcggaa ccggggttac agtgcctcaa ccctcccaga cgtagccaga gggaagcaac      360

tccctgatgc caaccgctgt gggctgccca tcggaatctt tgacaattgc cttgatcccc      420

gggtgcaagt caagcagcac ctgccgacat cgcccgcacg gagacagaat gccgcggttt      480

tcgttcccga tggccactat gcacgtcaga tttccggcag cagccgcagc ggccgttccg      540

aggaccacga gctccgcgca tggccctccg gtgaaatgat atacattcac gccggtaaag      600

atccgaccgt cggacgagag ggctgcactg gccaccgagt agtcctcgct aataggtatg      660

ctgttgatgg tcgcagttgc acgttcgatc agcgtggatt cctcttggga taaaggcttg      720

gccatcgagc tcggtacccg gggatccatg attgttgtat tatgtaccta tgtttgtgat      780

gagacaataa atatgagaag agaacgttgc ggccactttt ttctccttcc ttcgcgtgct      840

catgttggtg gtttgggagg cagaagatgc atggagcgcc acacattcgg taggacgaaa      900

cagcctcccc cacaaaggga ccatgggtag ctaggatgac gcacaagcga gttcccgctc      960

tcgaagggaa acccaggcat ttccttcctc ttttcaagcc acttgttcac gtgtcaacac     1020

aattttggac taaaatgccc ctcggaactc ggcaggcctc cctctgctcc gttgtcctgg     1080

tcgccgagaa cgcgagaccg tgccgcatgc catcgatctg ctcgtctgta ctactaatcg     1140

tgtgcgtgtt cgtgcttgtt tcgcacgaaa ttgtcctcgt tcggccctca caacggtgga     1200

aatcggtgct agaataaagt gaggtggctt atttcaatgg cggccgtcat catgcgggat     1260

caactgaagt acggcgggtt ctcgagattt catcgtgctc gtccagagca ggtgttttgc     1320

ctgcagctct tcatgtttag gggtcatgat ttcatctgat atgccgtaag aaaaccaata     1380

ttcacttctc aattttccat ggaaaggtga aggcctaggt tgtgtgcgag gcaacgactg     1440

gggagggatc gcaacattct tgctaacctc ccctctatct tggccgctgt gaatcggcat     1500

atttaccggg ctgaattgag aaagtgtttt gagggaatta aaaggtggct gtcttgcaag     1560

cttggcttca gtgcctgctt aattcgaacc gatccagctt gtgatgaggc cttcctaagc     1620

ctggtagtca gaagcgacat ggcgctataa atttcgtctc agttggagag tagaaaagca     1680

tgattcgaac acggttttca actgccaaag atatctccat tgtttccttc aatctgtaca     1740

cctgcacggt gcaccagttg gtacggcata ttatggttta ataagcatac atcatatgaa     1800

tacaattcag cttaaattta tcatacaaag atgtaagtgc agcgtgggtc tgtaacgatc     1860

gggcgtaatt taagataatg cgagggaccg ggggaggttt tggaacggaa tgaggaatgg     1920

gtcatggccc ataataataa tatgggtttg gtcgcctcgc acagcaaccg tacgtgcgaa     1980

aaaggaacag atccatttaa taagttgaac gttattcttt cctatgcaat gcgtgtatcg     2040

gaggcgagag caagtcatag gtggctgcgc acaataattg agtctcagct gagcgccgtc     2100

cgcgggtggt gtgagtggtc atcctcctcc cggcctatcg ctcacatcgc ctctcaatgg     2160

tggtggtggg gcctgatatg acctcaatgc cgacccatat taaaacccag taaagcattc     2220

accaacgaac gaggggctct tttgtgtgtg ttttgagtat gattttacac ctctttgtgc     2280

atctctctgg tcttccttgg ttcccgtagt ttgggcatca tcactcacgc ttccctcgac     2340

cttcgttctt cctttacaac cccgacacag gtcagagttg gagtaatcaa aaaaggggtg     2400

cacgaatgag atacattaga ttttgacaga tatcctttta ctggagaggg ttcaagggat     2460

caaatgaaca gcgggcgttg gcaatctagg gagggatcgg aggttggcag cgagcgaaag     2520

cgtgtccatc cttttggctg tcacacctca cgaaccaact gttagcaggc cagcacagat     2580

gacatacgag aatctttatt atatcgtaga ccttatgtgg atgacctttg gtgctgtgtg     2640

tctggcaatg aacctgaagg cttgataggg aggtggctcc cgtaaaccct ttgtcctttc     2700

cacgctgagt ctcccccgca ctgtccttta tacaaattgt tacagtcatc tgcaggcggt     2760

ttttctttgg caggcaaaga tgcccaagaa aaagcggaag gtcggcgact acaaggatga     2820

cgatgacaag ttggagcctg gagagaagcc ctacaaatgc cctgagtgcg gaaagagctt     2880

cagccaatct ggagccttga cccggcatca acgaacgcat acacgagaca agaagtactc     2940

catcgggctg gacatcggga cgaactccgt gggatgggcc gtgatcacag acgaatacaa     3000

ggtgccttcc aagaagttca aggtgctggg gaacacggac agacactcca tcaagaagaa     3060

cctcatcggg gccttgctct tcgactccgg agaaaccgcc gaagcaacgc gattgaaaag     3120

aaccgccaga agacgataca cacgacggaa gaaccgcatc tgctacctcc aggagatctt     3180

cagcaacgag atggccaagg tggacgactc gttctttcat cgcctggagg agagcttcct     3240

ggtggaggaa gacaagaaac atgagcgcca cccgatcttc gggaacatcg tggacgaagt     3300

ggcctaccac gagaaatacc ccacgatcta ccacttgcgc aagaaactcg tggactccac     3360

ggacaaagcg gacttgcggt tgatctactt ggccttggcc cacatgatca aatttcgggg     3420

ccacttcctg atcgagggcg acttgaatcc cgacaattcc gacgtggaca agctcttcat     3480

ccagctggtg cagacctaca accagctctt cgaggagaac cccatcaatg cctccggagt     3540

ggacgccaaa gccatcttgt ccgcccgatt gtccaaatcc agacgcttgg agaacttgat     3600

cgcacaactt cctggcgaga agaagaacgg cctcttcggc aacttgatcg cgctgtcgct     3660

gggattgacg cctaacttca agtccaactt cgacttggcc gaggacgcca agttgcaact     3720

gtccaaggac acctacgacg acgacctcga caacctgctg gcccaaattg gcgaccaata     3780

cgcggacttg tttttggcgg ccaagaactt gagcgacgcc atcttgttga gcgacatctt     3840

gcgcgtgaat acggagatca ccaaagcccc tttgtccgcc tctatgatca agcggtacga     3900

cgagcaccac caagacttga ccctgttgaa agccctcgtg cggcaacaat tgcccgagaa     3960

gtacaaggag atcttcttcg accagtccaa gaacgggtac gccggctaca tcgacggagg     4020

agcctcccaa gaagagttct acaagttcat caagcccatc ctggagaaga tggacggcac     4080

cgaggagttg ctcgtgaagc tgaaccgcga agacttgttg cgaaaacagc ggacgttcga     4140

caatggcagc atcccccacc aaatccattt gggagagttg cacgccatct tgcgacggca     4200

agaggacttc tacccgttcc tgaaggacaa ccgcgagaaa atcgagaaga tcctgacgtt     4260

cagaatcccc tactacgtgg gacccttggc ccgaggcaat tcccggtttg catggatgac     4320

gcgcaaaagc gaagagacga tcaccccctg gaacttcgaa gaagtggtcg acaaaggagc     4380

atccgcacag agcttcatcg agcgaatgac gaacttcgac aagaacctgc ccaacgagaa     4440

ggtgttgccc aagcattcgc tgctgtacga gtacttcacg gtgtacaacg agctgaccaa     4500

ggtgaagtac gtgaccgagg gcatgcgcaa acccgcgttc ctgtcgggag agcaaaagaa     4560

ggccattgtg gacctgctgt tcaagaccaa ccggaaggtg accgtgaaac agctgaaaga     4620

ggactacttc aagaagatcg agtgcttcga ctccgtggag atctccggcg tggaggaccg     4680

attcaatgcc tccttgggaa cctaccatga cctcctgaag atcatcaagg acaaggactt     4740

cctggacaac gaggagaacg aggacatcct ggaggacatc gtgctgaccc tgaccctgtt     4800

cgaggaccga gagatgatcg aggaacggtt gaaaacgtac gcccacttgt tcgacgacaa     4860

ggtgatgaag cagctgaaac gccgccgcta caccggatgg ggacgattga gccgcaaact     4920

gattaatgga attcgcgaca agcaatccgg aaagaccatc ctggacttcc tgaagtccga     4980

cgggttcgcc aaccgcaact tcatgcagct catccacgac gactccttga ccttcaagga     5040

ggacatccag aaggcccaag tgtccggaca aggagactcc ttgcacgagc acatcgccaa     5100

tttggccgga tcccccgcaa tcaaaaaagg catcttgcaa accgtgaaag tggtcgacga     5160

actggtgaag gtgatgggac ggcacaagcc cgagaacatc gtgatcgaaa tggcccgcga     5220

gaaccaaacc acccaaaaag gacagaagaa ctcccgagag cgcatgaagc ggatcgaaga     5280

gggcatcaag gagttgggct cccagatcct gaaggagcat cccgtggaga atacccaatt     5340

gcaaaacgag aagctctacc tctactacct ccagaacggg cgggacatgt acgtcgacca     5400

agagctggac atcaaccgcc tctccgacta cgatgtggat catattgtgc cccagagctt     5460

cctcaaggac gacagcatcg acaacaaggt cctgacgcgc agcgacaaga accggggcaa     5520

gtctgacaat gtgccttccg aagaagtcgt gaagaagatg aagaactact ggcggcagct     5580

gctcaacgcc aagctcatca cccaacggaa gttcgacaac ctgaccaagg ccgagagagg     5640

aggattgtcc gagttggaca aagccggctt cattaaacgc caactcgtgg agacccgcca     5700

gatcacgaag cacgtggccc aaatcttgga ctcccggatg aacacgaaat acgacgagaa     5760

tgacaagctg atccgcgagg tgaaggtgat cacgctgaag tccaagctgg tgagcgactt     5820

ccggaaggac ttccagttct acaaggtgcg ggagatcaac aactaccatc acgcccatga     5880

cgcctacctg aacgccgtgg tcggaaccgc cctgatcaag aaatacccca agctggagtc     5940

cgaattcgtg tacggagatt acaaggtcta cgacgtgcgg aagatgatcg cgaagtccga     6000

gcaggagatc ggcaaagcca ccgccaagta cttcttttac tccaacatca tgaacttctt     6060

caagaccgag atcacgctcg ccaacggcga gatccgcaag cgccccctga tcgagaccaa     6120

cggcgagacg ggagagattg tgtgggacaa aggaagagat tttgccacag tgcgcaaggt     6180

gctgtccatg cctcaggtga acatcgtgaa gaagaccgag gtgcaaacag gagggttttc     6240

caaagagtcc attttgccta agaggaattc cgacaagctc atcgcccgca agaaggactg     6300

ggaccccaag aagtacgggg gcttcgactc ccccacggtg gcctactccg tgttggtggt     6360

ggccaaagtg gagaaaggga agagcaagaa gctgaaatcc gtgaaggagt tgctcggaat     6420

cacgatcatg gaacgatcgt cgttcgagaa aaaccccatc gacttcctcg aagccaaagg     6480

gtacaaagag gtgaagaagg acctgatcat caagctgccc aagtactccc tgttcgagct     6540

ggagaacggc cgcaagcgga tgctggcctc cgccggggaa ctgcagaaag ggaacgaatt     6600

ggccttgccc tccaaatacg tgaacttcct ctacttggcc tcccattacg aaaagctcaa     6660

aggatcccct gaggacaatg agcagaagca actcttcgtg gaacaacaca agcactacct     6720

ggacgagatc atcgagcaga tcagcgagtt ctccaagcgc gtgatcctcg ccgacgccaa     6780

cctggacaag gtgctctccg cctacaacaa gcaccgcgac aagcctatcc gcgagcaagc     6840

cgagaatatc attcacctgt ttaccctgac gaatttggga gcccctgccg cctttaaata     6900

ctttgacacc accatcgacc gcaaaagata cacctccacc aaggaagtct tggacgccac     6960

cctcatccac cagtccatca cgggcctcta cgagacgcgc atcgacctct cccaattggg     7020

cggcgactaa agtgatgcgg cctttaggaa acaccacaaa agtaattgac aatctcagga     7080

acgatctgcg tgtttacagc ttcccaaata acaattatac cacgtaccaa aaggggttta     7140

atgtatctca caaattcttc taataggtac agcttctcaa attgggtgta tgatgtgaca     7200

cttcgtctca cacacgtcac gataattcag cgtatggctt cccttcatca cattcacgca     7260

aacttctaca caaccctggg catatttctt gtgttggcaa cactcccgaa atcgattctg     7320

cacacaatgg ttcattcaat gattcaagta cgttttagac ggactaggca gtttaattaa     7380

aaacatctat cctccagatc accagggcca gtgaggccgg cataaaggac ggcaaggaaa     7440

gaaaagaaag aaagaaaagg acacttatag catagtttga agttataagt agtcgcaatc     7500

tgtgtgcagc cgacagatgc tttttttttc cgtttggcag gaggtgtagg gatgtcgaag     7560

accagtccag ctagtatcta tcctacaagt caatcatgct gcgacaaaaa tttctcgcac     7620

gaggcctctc gataaacaaa actttaaaag cacacttcat tgtcatgcag agtaataact     7680

cttccgcgtc gatcaattta tcaatctcta tcatttccgc ccctttcctt gcatagagca     7740

agaaaagcga cccggatgag gataacatgt cctgcgccag tagtgtggca ttgcctgtct     7800

ctcatttaca cgtactgaaa gcataatgca cgcgcatacc aatatttttc gtgtacggag     7860

atgaagagac gcgacacgta agatcacgag aaggcgagca cggttgccaa tggcagacgc     7920

gctagtctcc attatcgcgt tgttcggtag cttgctgcat gtcttcagtg gcactatatc     7980

cactctgcct cgtcttctac acgagggcca catcggtgca agttcgaaaa atcatatctc     8040

aatcttcaga tcctttccag aaacggtgct caggcgggaa agtgaaggtt ttctactcta     8100

gtggctaccc caattctctc cgactgtcgc agacggtcct tcgttgcgca cgcaccgcgc     8160

actacctctg aaattcgaca accgaagttc aattttacat ctaacttctt tcccattctc     8220

tcaccaaaag cctagcttac atgttggaga gcgacgagag cggcctgccc gccatggaga     8280

tcgagtgccg catcaccggc accctgaacg gcgtggagtt cgagctggtg ggcggcggag     8340

agggcacccc cgagcagggc cgcatgacca acaagatgaa gagcaccaaa ggcgccctga     8400

ccttcagccc ctacctgctg agccacgtga tgggctacgg cttctaccac ttcggcacct     8460

accccagcgg ctacgagaac cccttcctgc acgccatcaa caacggcggc tacaccaaca     8520

cccgcatcga gaagtacgag gacggcggcg tgctgcacgt gagcttcagc taccgctacg     8580

aggccggccg cgtgatcggc gacttcaagg tgatgggcac cggcttcccc gaggacagcg     8640

tgatcttcac cgacaagatc atccgcagca acgccaccgt ggagcacctg caccccatgg     8700

gcgataacga tctggatggc agcttcaccc gcaccttcag cctgcgcgac ggcggctact     8760

acagctccgt ggtggacagc cacatgcact tcaagagcgc catccacccc agcatcctgc     8820

agaacggggg ccccatgttc gccttccgcc gcgtggagga ggatcacagc aacaccgagc     8880

tgggcatcgt ggagtaccag cacgccttca agaccccgga tgcagatgcc ggtgaagaat     8940

aagggtggga aggagtcggg gagggtcctg gcagagcggc gtcctcatga tgtgttggag     9000

acctggagag tcgagagctt cctcgtcacc tgattgtcat gtgtgtatag gttaaggggg     9060

cccactcaaa gccataaaga cgaacacaaa cactaatctc aacaaagtct actagcatgc     9120

cgtctgtcca tctttatttc ctggcgcgcc tatgcttgta aaccgttttg tgaaaaaatt     9180

tttaaaataa aaaaggggac ctctagggtc cccaattaat tagtaatata atctattaaa     9240

ggtcattcaa aaggtcatcc agacgaaagg gcctcgtgat acgcctattt ttataggtta     9300

atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg     9360

gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat     9420

aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc     9480

gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa     9540

cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac     9600

tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga     9660

tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag     9720

agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca     9780

cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca     9840

tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa     9900

ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc     9960

tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa    10020

cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag    10080

actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct    10140

ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac    10200

tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa    10260

ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt    10320

aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat    10380

ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg    10440

agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc    10500

ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg    10560

tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag    10620

cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact    10680

ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg    10740

gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc    10800

ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg    10860

aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg    10920

cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag    10980

ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc    11040

gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct    11100

ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc    11160

ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc    11220

gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga aga                      11263


<210>  7
<211>  4101
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic


<220>
<221>  misc_feature
<223>  Cas9 gene codon optimized for Nannochloropsis

<400>  7
gacaagaagt actccatcgg gctggacatc gggacgaact ccgtgggatg ggccgtgatc       60

acagacgaat acaaggtgcc ttccaagaag ttcaaggtgc tggggaacac ggacagacac      120

tccatcaaga agaacctcat cggggccttg ctcttcgact ccggagaaac cgccgaagca      180

acgcgattga aaagaaccgc cagaagacga tacacacgac ggaagaaccg catctgctac      240

ctccaggaga tcttcagcaa cgagatggcc aaggtggacg actcgttctt tcatcgcctg      300

gaggagagct tcctggtgga ggaagacaag aaacatgagc gccacccgat cttcgggaac      360

atcgtggacg aagtggccta ccacgagaaa taccccacga tctaccactt gcgcaagaaa      420

ctcgtggact ccacggacaa agcggacttg cggttgatct acttggcctt ggcccacatg      480

atcaaatttc ggggccactt cctgatcgag ggcgacttga atcccgacaa ttccgacgtg      540

gacaagctct tcatccagct ggtgcagacc tacaaccagc tcttcgagga gaaccccatc      600

aatgcctccg gagtggacgc caaagccatc ttgtccgccc gattgtccaa atccagacgc      660

ttggagaact tgatcgcaca acttcctggc gagaagaaga acggcctctt cggcaacttg      720

atcgcgctgt cgctgggatt gacgcctaac ttcaagtcca acttcgactt ggccgaggac      780

gccaagttgc aactgtccaa ggacacctac gacgacgacc tcgacaacct gctggcccaa      840

attggcgacc aatacgcgga cttgtttttg gcggccaaga acttgagcga cgccatcttg      900

ttgagcgaca tcttgcgcgt gaatacggag atcaccaaag cccctttgtc cgcctctatg      960

atcaagcggt acgacgagca ccaccaagac ttgaccctgt tgaaagccct cgtgcggcaa     1020

caattgcccg agaagtacaa ggagatcttc ttcgaccagt ccaagaacgg gtacgccggc     1080

tacatcgacg gaggagcctc ccaagaagag ttctacaagt tcatcaagcc catcctggag     1140

aagatggacg gcaccgagga gttgctcgtg aagctgaacc gcgaagactt gttgcgaaaa     1200

cagcggacgt tcgacaatgg cagcatcccc caccaaatcc atttgggaga gttgcacgcc     1260

atcttgcgac ggcaagagga cttctacccg ttcctgaagg acaaccgcga gaaaatcgag     1320

aagatcctga cgttcagaat cccctactac gtgggaccct tggcccgagg caattcccgg     1380

tttgcatgga tgacgcgcaa aagcgaagag acgatcaccc cctggaactt cgaagaagtg     1440

gtcgacaaag gagcatccgc acagagcttc atcgagcgaa tgacgaactt cgacaagaac     1500

ctgcccaacg agaaggtgtt gcccaagcat tcgctgctgt acgagtactt cacggtgtac     1560

aacgagctga ccaaggtgaa gtacgtgacc gagggcatgc gcaaacccgc gttcctgtcg     1620

ggagagcaaa agaaggccat tgtggacctg ctgttcaaga ccaaccggaa ggtgaccgtg     1680

aaacagctga aagaggacta cttcaagaag atcgagtgct tcgactccgt ggagatctcc     1740

ggcgtggagg accgattcaa tgcctccttg ggaacctacc atgacctcct gaagatcatc     1800

aaggacaagg acttcctgga caacgaggag aacgaggaca tcctggagga catcgtgctg     1860

accctgaccc tgttcgagga ccgagagatg atcgaggaac ggttgaaaac gtacgcccac     1920

ttgttcgacg acaaggtgat gaagcagctg aaacgccgcc gctacaccgg atggggacga     1980

ttgagccgca aactgattaa tggaattcgc gacaagcaat ccggaaagac catcctggac     2040

ttcctgaagt ccgacgggtt cgccaaccgc aacttcatgc agctcatcca cgacgactcc     2100

ttgaccttca aggaggacat ccagaaggcc caagtgtccg gacaaggaga ctccttgcac     2160

gagcacatcg ccaatttggc cggatccccc gcaatcaaaa aaggcatctt gcaaaccgtg     2220

aaagtggtcg acgaactggt gaaggtgatg ggacggcaca agcccgagaa catcgtgatc     2280

gaaatggccc gcgagaacca aaccacccaa aaaggacaga agaactcccg agagcgcatg     2340

aagcggatcg aagagggcat caaggagttg ggctcccaga tcctgaagga gcatcccgtg     2400

gagaataccc aattgcaaaa cgagaagctc tacctctact acctccagaa cgggcgggac     2460

atgtacgtcg accaagagct ggacatcaac cgcctctccg actacgatgt ggatcatatt     2520

gtgccccaga gcttcctcaa ggacgacagc atcgacaaca aggtcctgac gcgcagcgac     2580

aagaaccggg gcaagtctga caatgtgcct tccgaagaag tcgtgaagaa gatgaagaac     2640

tactggcggc agctgctcaa cgccaagctc atcacccaac ggaagttcga caacctgacc     2700

aaggccgaga gaggaggatt gtccgagttg gacaaagccg gcttcattaa acgccaactc     2760

gtggagaccc gccagatcac gaagcacgtg gcccaaatct tggactcccg gatgaacacg     2820

aaatacgacg agaatgacaa gctgatccgc gaggtgaagg tgatcacgct gaagtccaag     2880

ctggtgagcg acttccggaa ggacttccag ttctacaagg tgcgggagat caacaactac     2940

catcacgccc atgacgccta cctgaacgcc gtggtcggaa ccgccctgat caagaaatac     3000

cccaagctgg agtccgaatt cgtgtacgga gattacaagg tctacgacgt gcggaagatg     3060

atcgcgaagt ccgagcagga gatcggcaaa gccaccgcca agtacttctt ttactccaac     3120

atcatgaact tcttcaagac cgagatcacg ctcgccaacg gcgagatccg caagcgcccc     3180

ctgatcgaga ccaacggcga gacgggagag attgtgtggg acaaaggaag agattttgcc     3240

acagtgcgca aggtgctgtc catgcctcag gtgaacatcg tgaagaagac cgaggtgcaa     3300

acaggagggt tttccaaaga gtccattttg cctaagagga attccgacaa gctcatcgcc     3360

cgcaagaagg actgggaccc caagaagtac gggggcttcg actcccccac ggtggcctac     3420

tccgtgttgg tggtggccaa agtggagaaa gggaagagca agaagctgaa atccgtgaag     3480

gagttgctcg gaatcacgat catggaacga tcgtcgttcg agaaaaaccc catcgacttc     3540

ctcgaagcca aagggtacaa agaggtgaag aaggacctga tcatcaagct gcccaagtac     3600

tccctgttcg agctggagaa cggccgcaag cggatgctgg cctccgccgg ggaactgcag     3660

aaagggaacg aattggcctt gccctccaaa tacgtgaact tcctctactt ggcctcccat     3720

tacgaaaagc tcaaaggatc ccctgaggac aatgagcaga agcaactctt cgtggaacaa     3780

cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa gcgcgtgatc     3840

ctcgccgacg ccaacctgga caaggtgctc tccgcctaca acaagcaccg cgacaagcct     3900

atccgcgagc aagccgagaa tatcattcac ctgtttaccc tgacgaattt gggagcccct     3960

gccgccttta aatactttga caccaccatc gaccgcaaaa gatacacctc caccaaggaa     4020

gtcttggacg ccaccctcat ccaccagtcc atcacgggcc tctacgagac gcgcatcgac     4080

ctctcccaat tgggcggcga c                                               4101


<210>  8
<211>  24
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic


<220>
<221>  misc_feature
<223>  encodes Nuclear localization sequence

<400>  8
cccaagaaaa agcggaaggt cggc                                              24


<210>  9
<211>  24
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic


<220>
<221>  misc_feature
<223>  encodes FLAG tag

<400>  9
gactacaagg atgacgatga caag                                              24


<210>  10
<211>  147
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic


<220>
<221>  misc_feature
<223>  encodes Nuclear localization sequence-peptide linker-FLAG tag

<400>  10
atgcccaaga aaaagcggaa ggtcggcgac tacaaggatg acgatgacaa gttggagcct       60

ggagagaagc cctacaaatg ccctgagtgc ggaaagagct tcagccaatc tggagccttg      120

acccggcatc aacgaacgca tacacga                                          147


<210>  11
<211>  1000
<212>  DNA
<213>  Nannochloropsis gaditana


<220>
<221>  misc_feature
<223>  RPL24 promoter

<400>  11
aataagcata catcatatga atacaattca gcttaaattt atcatacaaa gatgtaagtg       60

cagcgtgggt ctgtaacgat cgggcgtaat ttaagataat gcgagggacc gggggaggtt      120

ttggaacgga atgaggaatg ggtcatggcc cataataata atatgggttt ggtcgcctcg      180

cacagcaacc gtacgtgcga aaaaggaaca gatccattta ataagttgaa cgttattctt      240

tcctatgcaa tgcgtgtatc ggaggcgaga gcaagtcata ggtggctgcg cacaataatt      300

gagtctcagc tgagcgccgt ccgcgggtgg tgtgagtggt catcctcctc ccggcctatc      360

gctcacatcg cctctcaatg gtggtggtgg ggcctgatat gacctcaatg ccgacccata      420

ttaaaaccca gtaaagcatt caccaacgaa cgaggggctc ttttgtgtgt gttttgagta      480

tgattttaca cctctttgtg catctctctg gtcttccttg gttcccgtag tttgggcatc      540

atcactcacg cttccctcga ccttcgttct tcctttacaa ccccgacaca ggtcagagtt      600

ggagtaatca aaaaaggggt gcacgaatga gatacattag attttgacag atatcctttt      660

actggagagg gttcaaggga tcaaatgaac agcgggcgtt ggcaatctag ggagggatcg      720

gaggttggca gcgagcgaaa gcgtgtccat ccttttggct gtcacacctc acgaaccaac      780

tgttagcagg ccagcacaga tgacatacga gaatctttat tatatcgtag accttatgtg      840

gatgaccttt ggtgctgtgt gtctggcaat gaacctgaag gcttgatagg gaggtggctc      900

ccgtaaaccc tttgtccttt ccacgctgag tctcccccgc actgtccttt atacaaattg      960

ttacagtcat ctgcaggcgg tttttctttg gcaggcaaag                           1000


<210>  12
<211>  317
<212>  DNA
<213>  Nannochloropsis gaditana


<220>
<221>  misc_feature
<223>  Bidirectional terminator 2

<400>  12
agtgatgcgg cctttaggaa acaccacaaa agtaattgac aatctcagga acgatctgcg       60

tgtttacagc ttcccaaata acaattatac cacgtaccaa aaggggttta atgtatctca      120

caaattcttc taataggtac agcttctcaa attgggtgta tgatgtgaca cttcgtctca      180

cacacgtcac gataattcag cgtatggctt cccttcatca cattcacgca aacttctaca      240

caaccctggg catatttctt gtgttggcaa cactcccgaa atcgattctg cacacaatgg      300

ttcattcaat gattcaa                                                     317


<210>  13
<211>  399
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic


<220>
<221>  misc_feature
<223>  blasticidin S deaminase gene from Aspergillus terreus codon 
       optimized for N. gaditana

<400>  13
atggccaagc ctttatccca agaggaatcc acgctgatcg aacgtgcaac tgcgaccatc       60

aacagcatac ctattagcga ggactactcg gtggccagtg cagccctctc gtccgacggt      120

cggatcttta ccggcgtgaa tgtatatcat ttcaccggag ggccatgcgc ggagctcgtg      180

gtcctcggaa cggccgctgc ggctgctgcc ggaaatctga cgtgcatagt ggccatcggg      240

aacgaaaacc gcggcattct gtctccgtgc gggcgatgtc ggcaggtgct gcttgacttg      300

cacccgggga tcaaggcaat tgtcaaagat tccgatgggc agcccacagc ggttggcatc      360

agggagttgc ttccctctgg ctacgtctgg gagggttga                             399


<210>  14
<211>  999
<212>  DNA
<213>  Nannochloropsis gaditana


<220>
<221>  misc_feature
<223>  TCTP promoter

<400>  14
cgtgcaggtg tacagattga aggaaacaat ggagatatct ttggcagttg aaaaccgtgt       60

tcgaatcatg cttttctact ctccaactga gacgaaattt atagcgccat gtcgcttctg      120

actaccaggc ttaggaaggc ctcatcacaa gctggatcgg ttcgaattaa gcaggcactg      180

aagccaagct tgcaagacag ccacctttta attccctcaa aacactttct caattcagcc      240

cggtaaatat gccgattcac agcggccaag atagagggga ggttagcaag aatgttgcga      300

tccctcccca gtcgttgcct cgcacacaac ctaggccttc acctttccat ggaaaattga      360

gaagtgaata ttggttttct tacggcatat cagatgaaat catgacccct aaacatgaag      420

agctgcaggc aaaacacctg ctctggacga gcacgatgaa atctcgagaa cccgccgtac      480

ttcagttgat cccgcatgat gacggccgcc attgaaataa gccacctcac tttattctag      540

caccgatttc caccgttgtg agggccgaac gaggacaatt tcgtgcgaaa caagcacgaa      600

cacgcacacg attagtagta cagacgagca gatcgatggc atgcggcacg gtctcgcgtt      660

ctcggcgacc aggacaacgg agcagaggga ggcctgccga gttccgaggg gcattttagt      720

ccaaaattgt gttgacacgt gaacaagtgg cttgaaaaga ggaaggaaat gcctgggttt      780

cccttcgaga gcgggaactc gcttgtgcgt catcctagct acccatggtc cctttgtggg      840

ggaggctgtt tcgtcctacc gaatgtgtgg cgctccatgc atcttctgcc tcccaaacca      900

ccaacatgag cacgcgaagg aaggagaaaa aagtggccgc aacgttctct tctcatattt      960

attgtctcat cacaaacata ggtacataat acaacaatc                             999


<210>  15
<211>  318
<212>  DNA
<213>  Nannochloropsis gaditana


<220>
<221>  misc_feature
<223>  EIF3 terminator

<400>  15
ggcactgtaa ccccggttcc gctcgacgaa ggctgggagc gccctttcgg tgggataaaa       60

tggatgcttt accgctgcgc ttcggctgag gaagagagaa atgcgagcgg ggatcggggt      120

cctagaaacg aagaaaggag aacaagttcc tggccaaaga aaaacaagac aaataccctc      180

tccaggcctg ggcccattac ttttttttgc tgtttcttat acctgcactc gtgcttctct      240

agtctgtcga gaccttacct gatcttcctc cctccatcgc tccccgcccc ccccatccga      300

gcaaccgtcg accatacg                                                    318


<210>  16
<211>  702
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic


<220>
<221>  misc_feature
<223>  Turbo GFP gene codon optimized for Nannochloropsis

<400>  16
atgttggaga gcgacgagag cggcctgccc gccatggaga tcgagtgccg catcaccggc       60

accctgaacg gcgtggagtt cgagctggtg ggcggcggag agggcacccc cgagcagggc      120

cgcatgacca acaagatgaa gagcaccaaa ggcgccctga ccttcagccc ctacctgctg      180

agccacgtga tgggctacgg cttctaccac ttcggcacct accccagcgg ctacgagaac      240

cccttcctgc acgccatcaa caacggcggc tacaccaaca cccgcatcga gaagtacgag      300

gacggcggcg tgctgcacgt gagcttcagc taccgctacg aggccggccg cgtgatcggc      360

gacttcaagg tgatgggcac cggcttcccc gaggacagcg tgatcttcac cgacaagatc      420

atccgcagca acgccaccgt ggagcacctg caccccatgg gcgataacga tctggatggc      480

agcttcaccc gcaccttcag cctgcgcgac ggcggctact acagctccgt ggtggacagc      540

cacatgcact tcaagagcgc catccacccc agcatcctgc agaacggggg ccccatgttc      600

gccttccgcc gcgtggagga ggatcacagc aacaccgagc tgggcatcgt ggagtaccag      660

cacgccttca agaccccgga tgcagatgcc ggtgaagaat aa                         702


<210>  17
<211>  822
<212>  DNA
<213>  Nannochloropsis gaditana


<220>
<221>  misc_feature
<223>  4A-III promoter

<400>  17
ggcataaagg acggcaagga aagaaaagaa agaaagaaaa ggacacttat agcatagttt       60

gaagttataa gtagtcgcaa tctgtgtgca gccgacagat gctttttttt tccgtttggc      120

aggaggtgta gggatgtcga agaccagtcc agctagtatc tatcctacaa gtcaatcatg      180

ctgcgacaaa aatttctcgc acgaggcctc tcgataaaca aaactttaaa agcacacttc      240

attgtcatgc agagtaataa ctcttccgcg tcgatcaatt tatcaatctc tatcatttcc      300

gcccctttcc ttgcatagag caagaaaagc gacccggatg aggataacat gtcctgcgcc      360

agtagtgtgg cattgcctgt ctctcattta cacgtactga aagcataatg cacgcgcata      420

ccaatatttt tcgtgtacgg agatgaagag acgcgacacg taagatcacg agaaggcgag      480

cacggttgcc aatggcagac gcgctagtct ccattatcgc gttgttcggt agcttgctgc      540

atgtcttcag tggcactata tccactctgc ctcgtcttct acacgagggc cacatcggtg      600

caagttcgaa aaatcatatc tcaatcttca gatcctttcc agaaacggtg ctcaggcggg      660

aaagtgaagg ttttctactc tagtggctac cccaattctc tccgactgtc gcagacggtc      720

cttcgttgcg cacgcaccgc gcactacctc tgaaattcga caaccgaagt tcaattttac      780

atctaacttc tttcccattc tctcaccaaa agcctagctt ac                         822


<210>  18
<211>  200
<212>  DNA
<213>  Nannochloropsis gaditana


<220>
<221>  misc_feature
<223>  Bidirectional terminator 5

<400>  18
gggtgggaag gagtcgggga gggtcctggc agagcggcgt cctcatgatg tgttggagac       60

ctggagagtc gagagcttcc tcgtcacctg attgtcatgt gtgtataggt taagggggcc      120

cactcaaagc cataaagacg aacacaaaca ctaatctcaa caaagtctac tagcatgccg      180

tctgtccatc tttatttcct                                                  200


<210>  19
<211>  1029
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic


<220>
<221>  misc_feature
<223>  Hyg resistance gene

<400>  19
atggggaaga aaccggaact gaccgctacg tccgtggaga aattccttat tgagaagttc       60

gactctgtct ccgacttgat gcaactgagc gagggagagg agagtagggc gttctcgttt      120

gacgtagggg gtcggggata cgtgttgagg gttaatagtt gtgcggacgg gttctacaag      180

gatcggtatg tctaccgtca tttcgcctcc gccgctctcc ccataccaga ggtactggac      240

attggggagt ttagcgaatc tctcacgtac tgcatctcgc gccgagccca gggagtgacg      300

ttgcaagatc tgcccgaaac tgaattgcct gccgttttgc aacccgtggc cgaggccatg      360

gacgcgatcg ctgccgcaga tctgtctcag acgtccggct ttggaccttt tgggccccag      420

ggcatcgggc agtacacgac ctggcgagac ttcatctgcg ccattgccga tcctcacgtc      480

tatcattggc agacagtcat ggatgacacc gtgtctgcat ccgtggccca agcactggac      540

gaactcatgt tgtgggccga ggattgccct gaggtcaggc acctggtgca cgcggatttc      600

ggcagcaata acgtacttac agacaatggt cggattactg ctgtcatcga ctggtccgaa      660

gcgatgtttg gtgatagcca atacgaagtg gcgaacatat tcttctggcg tccctggttg      720

gcgtgcatgg agcagcagac acgctacttt gaacggaggc acccggagct ggccggctcc      780

ccacgactcc gcgcctatat gttgcgtatc ggactcgatc agctttacca gtctctcgtc      840

gacggcaact tcgacgacgc cgcgtgggcg cagggccgct gcgacgcgat agtccgcagc      900

ggggctggga cggtgggtcg gacccaaatc gcacgccggt cggctgcggt gtggacagac      960

ggctgtgttg aggtgcttgc ggactcgggc aaccgtaggc cgagcacccg accgcgtgca     1020

aaggagtga                                                             1029


<210>  20
<211>  1000
<212>  DNA
<213>  Nannochloropsis gaditana


<220>
<221>  misc_feature
<223>  EIF3 promoter

<400>  20
tcataatcaa agatgagcca gccacgaagc taccggagaa ttctgtaaga aaaatgttta       60

aagttgaaaa tgctaacagt gaagtgatat ccttttttaa tggagtgttg aggtgaagtc      120

tagcatcgta ggggaaaaca ggattctgtg tcttccattc tactccttga taaagcgaag      180

aaatccgaca aaaccaaaga gattgttcaa gtttaagatt tgtaagcgta caactatgaa      240

cttcttctct ttgtaggcct gagtggtcgt atgcatacga ttcatgaagt gaatcagtat      300

cgctggattt tgcttaggag taaagcacaa ctaagaaaat atgctgcctg gcaggcatcc      360

tgagacatga ggcaagcgac gtagcaattg aatcctaatt taagccaggg catctgtatg      420

actctgttag ttaattgatg aaccaatgag ctttaaaaaa aaatcgttgc gcgtaatgta      480

gttttaattc tccgccttga ggtgcggggc catttcggac aaggttcttt ggacggagat      540

ggcagcatgt gtcccttctc caaattggtc cgtgtggtag ttgagatgct gccttaaaat      600

tctgctcggt catcctgcct tcgcattcac tcctttcgag ctgtcgggtt cctcacgagg      660

cctccgggag cggattgcgc agaaaggcga cccggagaca cagagaccat acaccgacta      720

aattgcactg gacgatacgg catggcgacg acgatggcca agcattgcta cgtgattatt      780

cgccttgtca ttcagggaga aatgatgaca tgtgtgggac ggtctttaca tgggaagagg      840

gcatgaaaat aacatggcct ggcgggatgg agcgtcacac ctgtgtatgc gttcgatcca      900

caagcaactc accatttgcg tcggggcctg tctccaatct gctttaggct acttttctct      960

aatttagcct attctataca gacagagaca cacagggatc                           1000


<210>  21
<211>  27
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic


<220>
<221>  misc_feature
<223>  27 nucleotide 5' ID sequence

<400>  21
tccacagccc gaacccatga gagagaa                                           27


<210>  22
<211>  27
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic


<220>
<221>  misc_feature
<223>  27 nucleotide 3' ID sequence

<400>  22
gcccgaatcg agttgatggc ccgcaaa                                           27


<210>  23
<211>  2400
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic


<220>
<221>  misc_feature
<223>  HygR cassette with flanking ID sequences

<400>  23
tccacagccc gaacccatga gagagaatca taatcaaaga tgagccagcc acgaagctac       60

cggagaattc tgtaagaaaa atgtttaaag ttgaaaatgc taacagtgaa gtgatatcct      120

tttttaatgg agtgttgagg tgaagtctag catcgtaggg gaaaacagga ttctgtgtct      180

tccattctac tccttgataa agcgaagaaa tccgacaaaa ccaaagagat tgttcaagtt      240

taagatttgt aagcgtacaa ctatgaactt cttctctttg taggcctgag tggtcgtatg      300

catacgattc atgaagtgaa tcagtatcgc tggattttgc ttaggagtaa agcacaacta      360

agaaaatatg ctgcctggca ggcatcctga gacatgaggc aagcgacgta gcaattgaat      420

cctaatttaa gccagggcat ctgtatgact ctgttagtta attgatgaac caatgagctt      480

taaaaaaaaa tcgttgcgcg taatgtagtt ttaattctcc gccttgaggt gcggggccat      540

ttcggacaag gttctttgga cggagatggc agcatgtgtc ccttctccaa attggtccgt      600

gtggtagttg agatgctgcc ttaaaattct gctcggtcat cctgccttcg cattcactcc      660

tttcgagctg tcgggttcct cacgaggcct ccgggagcgg attgcgcaga aaggcgaccc      720

ggagacacag agaccataca ccgactaaat tgcactggac gatacggcat ggcgacgacg      780

atggccaagc attgctacgt gattattcgc cttgtcattc agggagaaat gatgacatgt      840

gtgggacggt ctttacatgg gaagagggca tgaaaataac atggcctggc gggatggagc      900

gtcacacctg tgtatgcgtt cgatccacaa gcaactcacc atttgcgtcg gggcctgtct      960

ccaatctgct ttaggctact tttctctaat ttagcctatt ctatacagac agagacacac     1020

agggatcatg gggaagaaac cggaactgac cgctacgtcc gtggagaaat tccttattga     1080

gaagttcgac tctgtctccg acttgatgca actgagcgag ggagaggaga gtagggcgtt     1140

ctcgtttgac gtagggggtc ggggatacgt gttgagggtt aatagttgtg cggacgggtt     1200

ctacaaggat cggtatgtct accgtcattt cgcctccgcc gctctcccca taccagaggt     1260

actggacatt ggggagttta gcgaatctct cacgtactgc atctcgcgcc gagcccaggg     1320

agtgacgttg caagatctgc ccgaaactga attgcctgcc gttttgcaac ccgtggccga     1380

ggccatggac gcgatcgctg ccgcagatct gtctcagacg tccggctttg gaccttttgg     1440

gccccagggc atcgggcagt acacgacctg gcgagacttc atctgcgcca ttgccgatcc     1500

tcacgtctat cattggcaga cagtcatgga tgacaccgtg tctgcatccg tggcccaagc     1560

actggacgaa ctcatgttgt gggccgagga ttgccctgag gtcaggcacc tggtgcacgc     1620

ggatttcggc agcaataacg tacttacaga caatggtcgg attactgctg tcatcgactg     1680

gtccgaagcg atgtttggtg atagccaata cgaagtggcg aacatattct tctggcgtcc     1740

ctggttggcg tgcatggagc agcagacacg ctactttgaa cggaggcacc cggagctggc     1800

cggctcccca cgactccgcg cctatatgtt gcgtatcgga ctcgatcagc tttaccagtc     1860

tctcgtcgac ggcaacttcg acgacgccgc gtgggcgcag ggccgctgcg acgcgatagt     1920

ccgcagcggg gctgggacgg tgggtcggac ccaaatcgca cgccggtcgg ctgcggtgtg     1980

gacagacggc tgtgttgagg tgcttgcgga ctcgggcaac cgtaggccga gcacccgacc     2040

gcgtgcaaag gagtgattga atcattgaat gaaccattgt gtgcagaatc gatttcggga     2100

gtgttgccaa cacaagaaat atgcccaggg ttgtgtagaa gtttgcgtga atgtgatgaa     2160

gggaagccat acgctgaatt atcgtgacgt gtgtgagacg aagtgtcaca tcatacaccc     2220

aatttgagaa gctgtaccta ttagaagaat ttgtgagata cattaaaccc cttttggtac     2280

gtggtataat tgttatttgg gaagctgtaa acacgcagat cgttcctgag attgtcaatt     2340

acttttgtgg tgtttcctaa aggccgcatc actgcccgaa tcgagttgat ggcccgcaaa     2400


<210>  24
<211>  20
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic


<220>
<221>  misc_feature
<223>  Forward primer to detect insertion of the donor fragment into the
       targeted locus of Naga_100148g8

<400>  24
accctgtcgc acatcctcct                                                   20


<210>  25
<211>  22
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic


<220>
<221>  misc_feature
<223>  Reverse primer to detect insertion of the donor fragment into the
       targeted locus of Naga_100148g8

<400>  25
ttctgaagac cgtggtccca gc                                                22


