                         SEQUENCE LISTING

<110>  SYNTHETIC GENOMICS, INC.
       MOELLERING, Eric R.
       EDWARDS, Amanda R.
       BAUMAN, Nicholas
 
<120>  REGULATORY ELEMENTS AND USES THEREOF

<130>  SGI1990-3WO

<150>  US 62/261,217
<151>  2015-11-30

<150>  US 62/261,777
<151>  2015-12-01

<150>  US 62/323,480
<151>  2016-04-15

<160>  51    

<170>  PatentIn version 3.5

<210>  1
<211>  530
<212>  DNA
<213>  Parachlorella sp.


<220>
<221>  misc_feature
<223>  RPS4 promoter

<400>  1
ccaccatggg ggaggtttga agtgtgcgcc tgatataatc atacacctaa aagcaccact       60

tgctgattgt gaagggacta tgtcgtttat gacgggacgt tacgctggcc gatggtttga      120

atttggacgc tgtggtagaa tgttatatgg acgtaaaggt tggcatattg aaaatcgtct      180

tcgcaggcaa acttctagac gtgtgaccca ccggtaaaac gacaagcgtg gcgcgtcgat      240

tgcgctttga acgtcgtttg ttggactcca gatgaacctc aaaatcaaag cggtgattga      300

cgaaaatcaa atgacagccc gcaaaatttc atcagccttc ggatcggatt ctcagaatct      360

gattgtccct gctggctaca tttatgaaat ttcgtacatt ttggcagaaa tgtcccaata      420

ccatagcact gccgcctgag ctcacccgag caatgcatac tgggtacctc gcccatctcg      480

ccctctttcc aagcccagtg ctgttgtaat agccaaaggg ctcagtaaca                 530


<210>  2
<211>  375
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic


<220>
<221>  misc_feature
<223>  Bleomycin resistance gene, codon-optimized for Parachlorella

<400>  2
atggccaaac tgacatccgc tgttcctgtg ttgacagcaa gagatgttgc aggtgcagtg       60

gagttttgga cagatagact ggggtttagc agggactttg tggaggacga ttttgcagga      120

gtggtgaggg atgatgtgac actgtttatc tcagcagtgc aggatcaagt ggtgcccgat      180

aatacactgg catgggtttg ggtgagagga ttggatgaac tgtatgcaga gtggtctgaa      240

gtggtgagca ccaactttag ggatgcaagc ggacctgcaa tgacagagat tggagaacaa      300

ccttggggaa gggagtttgc attgagagat cctgcaggga attgcgtgca ctttgttgca      360

gaagaacagg actga                                                       375


<210>  3
<211>  111
<212>  DNA
<213>  Nannochloropsis gaditana


<220>
<221>  misc_feature
<223>  T4 terminator

<400>  3
tggatgacct tttgaatgac ctttaataga ttatattact aattaattgg ggaccctaga       60

ggtccccttt tttattttaa aaattttttc acaaaacggt ttacaagcat a               111


<210>  4
<211>  1000
<212>  DNA
<213>  Parachlorella sp.


<220>
<221>  misc_feature
<223>  PsbR promoter

<400>  4
aagttgagca aggatgtggt ttgctgtttt agtcggggcc acagccttct cataggggtt       60

tgtttttgtg ctgggtactg tgacaattca tgtccttgtt cgcccgcatc tctgtttctg      120

gcgacctgtg ttaggctggc agagtacctc aagaacagca gggtgatcgc tttttccact      180

ttttcaataa actagtgtgc aagctaagta ggtacttggc agccagcggt caaatggtga      240

gcagattcat catattctaa gaatctcagc aattcaaaca tgcgtatgaa tcaacaacac      300

acgacttatt gcttatgcca agctactgca gatttcgaac aaatactcgt ctctgcttga      360

acaagtactt ttactcttgc aaaaaaacca ttcatgtttc tgtgatcata accgtgtgca      420

ccccgacagt acagacgcag ttatagtgac gtagatctgc acaggtaaac gattatgaca      480

ggctgccttt gagacgtggg tggatcacaa aagttgtgtc ttgcggtaga ttgatgctgt      540

caaaaaccaa aacttcaaag ccacaacgtt tgcaatgaat acctgataac aagccaacaa      600

tttgtgtctg accttgtgta catcgaaggt tcaaagagga cttcacagaa cttagtaaat      660

gaaactggcg tattgcctac agacaaggtt gtaagacaat cttcgggcgt cttttctcaa      720

gctaggatgt tcgataatga acaattaggg tttatattag attctagaag atattagaag      780

aaactgcaac cagccgcaga gggtttccgc tatatccgcc gttcaaagtt ttggcgcggt      840

gctattgtca ttccaccgca gtgtttgacc cggggacctc atccaatcgc tggcgtatcc      900

cactggcgcc agaaattctg aacaaacatg ctcactccgc agttgtgacc atctgttttc      960

ctgataaaac cagctgcgtt gtcattgtaa gcagatcata                           1000


<210>  5
<211>  702
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic


<220>
<221>  misc_feature
<223>  TurboGFP coding sequence, codon-optimized for Parachlorella

<400>  5
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>  6
<211>  200
<212>  DNA
<213>  Nannochloropsis gaditana


<220>
<221>  misc_feature
<223>  T5 terminator

<400>  6
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>  7
<211>  546
<212>  DNA
<213>  Parachlorella sp.


<220>
<221>  misc_feature
<223>  RSP4 Terminator

<400>  7
gcatagcatc agcctgtggc agggttgtgg tagggctgag tggcagggtt aaaggggttg       60

cctaccccac ccctactctc atgacaccag caacagcagc agctcatgca gtactcaaat      120

cactgatgtc aatggtgtga cacatttggt taaggctgct ttttaaagtg ctgctttggg      180

ggcagtgact gtgcagagct tggagcgtat ccccatgtaa tcagaaccga cgagagttcg      240

gggcaacctt tcatcttcac attttttgtg atcagctaca gagtctgaaa tcaaatagag      300

gctgccatct aaacgcagga gtcacaacga aggcgaaaac tccaattgct gtactcaatg      360

cactaagtga ttgttcaatg gataaataca ctatgctcaa ttcatgccag cagagctgct      420

ccttccagcc agctacaatg gctttttcca cgccttttga agtatgaatg ttcagcttgc      480

tgtgcttgat gcatcaccat aaacacaatt ctacaacatt tcatgccaac aacagtacgg      540

gctttc                                                                 546


<210>  8
<211>  572
<212>  DNA
<213>  Parachlorella sp.


<220>
<221>  misc_feature
<223>  ACP1 Promoter

<400>  8
agtttgcata gttaagtatg ctggctattg cagtacctta tatgcaaaca agtgctcaat       60

ctgtttcatc attgtctgtg ggcaaattgc ctgccaatat tctccagtta ttgcctgttg      120

tttcaaatga ttgaaattgg aagttgtatt gctctacatt tttgacttgt gattttttca      180

tttgttgata tctgacaact gtgaactgca ctgaacttgc tgtgcttata aatgcatttt      240

tttgttttgg gccacgttga ttccttgtga tactttcctg ctatcaaacc aaaaatatac      300

tctcatgact gacgtgcaac aaatgcatgg aagctttcaa cgttacgaca gctgcttgcc      360

ccccatcagc tattctacat gtgtaaccta ccttgcatgg ccaccacaac gctactgcat      420

gcaagatctg gcgcaactgg atgtcccaat agtagaagta tccggattat ctccgagagt      480

tttacatatg taatcgacgc catttctgtc atcaactata aatccattgc tcctgcattt      540

ctggcactga cattctacca caagcaatac ca                                    572


<210>  9
<211>  869
<212>  DNA
<213>  Parachlorella sp.


<220>
<221>  misc_feature
<223>  ACP1 Terminator

<400>  9
gcagcagctt gttatgcctt ccccatgggc atcagcatgc tgcaagctgt ctagatatcc       60

agctttcagt ggaggttgag cgagggtcag cagcggttcc ctggcgatgg cggtcagctt      120

ttctggaagc cttcactagg actgcgccca gcgcatgtga cgccaatcga acttgtgtgc      180

aaggccaaat tttgtgaccc tgtgctgcac ttcatgtatt caagaattga gaagaaattt      240

cattgctgcc cttctttcac tttaatttcc atccctggat ccacctccca ccattgtggt      300

tgatgggtag gggttttggg taggtgcagt tcgttgtgca cgttgacatg tgtaacggtg      360

agcaaaggaa ttgctgggca agtagctatt gcagcttaag ggcatggtga aacacttgtg      420

ctgtatttac agaggaagcc agacaggtaa ggagtgtgtg gcagcttgga acaggagggc      480

tggtcgcaac aagtatgcat atcccatgat tgttgacata agagcagcag gtgcatattg      540

ccagcctttg tgaaagtgga ttgaaaatcg attagttggt gtgatagctg aggctaggca      600

ctgccaacct gcagtgaaat gaggctccaa gaccgggtaa taatacaggc aatcgaatcc      660

agttgaaatt acggcgatta aatccaagcg agcgttgtaa gaacatctgc acctgtctga      720

agtagtgagc ggataatgag cattgcttgc cttctatcac tatacctgac agttacgtgt      780

cacacactct caagcacaac acacagcggc aaagttactt gctaaacctc acagtcaagc      840

tgaaaataaa ggctaaatta cgtgagacc                                        869


<210>  10
<211>  1707
<212>  DNA
<213>  Parachlorella sp.


<220>
<221>  misc_feature
<223>  encodes cpSRP54

<400>  10
atgcttcggc agcagctgtt gcacagcggc aggcagccgg gtgcgacatg cagcttacta       60

acctgctcga catggcgacc gtctgccttg ttcggccgtc ctaagcccca aaaactgcac      120

agccagcgct tgcagcatca gggccgcccc tcccgcctcg tcgtgcgcag cgcaatgttc      180

gacaacctga gccgcagcct ggagagggcg tgggacatgg tgcgcaagga cgggcggcta      240

acggcggaca acatcaagga gcccatgcgg gagattcgca gggcgctgct tgaggcggat      300

gtgaggctgg gggcgccgct gatcagattc ttggtatcta cccccccccc ctcccaggtc      360

tccctccccg tggtgcgcaa gtttgtgaag gcggtggagg agaaggcgct gggttctgca      420

gtgaccaagg gtgtcacccc cgaccagcag ctggtgaagg tggtgtacga ccagctgcgg      480

gagctgatgg gggggcagca ggaagggctg gtgcccactt cgccagagga gccgcaggtg      540

atcttgatgg cggggctgca gggcacgggg aagacgacag ctgcggggaa gctggccttg      600

ttcctgcaga agaaggggca gaaggtgctg ctggtggcca ccgacatcta ccgccccgcc      660

gccatcgacc agctggtgaa gctgggcgac aggatagggg tgccggtgtt ccagctggga      720

acccaggtgc agccgccgga gattgcaagg caggggctgg agaaggcgcg agcagagggg      780

tttgacgccg tcatcgtcga cacggcgggg cggctgcaga tcgaccagag catgatggag      840

gagctggtgc agatcaagtc cacggtgaag ccctccgaca cgctgctagt ggtcgatgcg      900

atgacggggc aggaggcagc cgggctggtg aaggcgttca atgatgccgt ggacatcaca      960

ggcgccgtgc tgaccaagct tgacggggac agccgcggcg gcgccgcgct gagcgtgcgc     1020

caggtcagcg ggcggcccat caagtttgtg ggcatggggg agggcatgga ggcgctggag     1080

cccttctacc ccgagcgcat ggccagcagg attctgggca tgggtgacgt ggtcaccctg     1140

gtggagaagg ctgaggagag catcaaggaa gaggaggcgc aggagatatc gcggaagatg     1200

ctgtcggcca aatttgactt tgacgacttc ctgaagcagt acaagatggt ggcggggatg     1260

gggaacatgg cccaaatcat gaagatgctg ccaggcatga acaagtttac ggagaagcag     1320

ctggcgggcg ttgagaagca gtacaaggtg tacgagagca tgatccagag catgacggtg     1380

aaggagcgca agcagccgga gctgttggtg aagtcgccct ccaggaggcg gcgcatagcg     1440

cgcgggtcgg ggcgctcgga gcgggaggtc acagagctgc tgggggtgtt caccaacctg     1500

cggacgcaga tgcagagctt ctccaaaatg atggccatgg gggggatggg catgggctcc     1560

atgatgagcg acgaggagat gatgcaggcc acgctggcag gcgccggccc ccgccccgtg     1620

ccagctggca aggtgcggcg gaagaagctg gccgcggcgg gcgggtcgcg gggcatggct     1680

gagctggcat ccctgaaggc agaatga                                         1707


<210>  11
<211>  1044
<212>  DNA
<213>  Parachlorella sp.


<220>
<221>  misc_feature
<223>  OCP-A Promoter

<400>  11
tgtgcatgta gatggcaagt taaagaagtg gtatacctgc aagtcatgtg tgcaacgtat       60

agagggtgtc atcccctcat gatcaacatg cgcggtaagg ttgcaaatca gacgggaggc      120

tgttactggt atgcaacgtt tcatcaaacc ataagatgtg tggggtgaaa tgcatactgg      180

ttttcagaat gatagtccag actggagtat ataacaatac acatcattgc attcaaatac      240

aaatatgttt atggtgagta tgcccgtgca cgagtgtcag agttacgctg ggactgaatt      300

tatcacacag cgcagcagtg ccctcccatc accggaagtc atttcatata aataccatat      360

acctatgtca acccgccaga accgtgtacg cactgttcca gatcatgcag ctgcttcaag      420

catagatgaa tctttcagca tcaactgttg ttaggtgaag accagctacc tacttcgcac      480

gcacaaaaat tgcacctgca ttcctgtgtc atctaaatct ctcctattga tgcctggtag      540

ctctgaattg attggaaact catactgtgt aatctaatat gtctgaaaca aagatgaatc      600

cacttgtttg aagcaaacag gaccgctgtg ctgtgcattg cacgtgagga ttgggtgtga      660

tcactttcat caaatcacgt tttttgtttt gcttgcagct tcacatcttt tcccctgcta      720

accacacacc cggaacaaaa acaaagccgt tatgtttgta tactccaaag cacggtcagt      780

cgtgatgctt taaggacaat aagtgcatat aatttgcaac aatagaaatt atgcatgtta      840

cagttgtatc gccctgtgtt cctcccatga cagccttgca aaaacttccc aatcgtgtgg      900

cagctgatga ggtggccagc tgctttgcat gcttcgcccg aggtttcact gaggatgata      960

cacagaaatt caggagaatg acacattgtt gattccaact ggctccctca acatcgtatc     1020

gtcagttcca ttggtacccg ccat                                            1044


<210>  12
<211>  598
<212>  DNA
<213>  Parachlorella sp.


<220>
<221>  misc_feature
<223>  OCP-A Terminator

<400>  12
ggaggaggac ctggtccccc tgctcgaaaa gcaggatttg ggcgggaggg tgcagggtct       60

gcagctaacc ggcagggccc tgcgcatctc agggtctcag ctcagctccc gcacagggtg      120

cagtgcagtg agatcagggc tgctggcccc gactgcagac gtgtgtacgg ctgcgtgcgt      180

aaccctgcgt catgcggcac agcattgcat ggctcccagt aggggacagc tgctgtgagt      240

ggagcccccc cccccctcca catcttgttg aggaacgcgc cgggatggtt gcatgcagcc      300

aggggtggca taaataggcc ttgattgttc ggctgccgat atgtactgcc atgtaaggtt      360

gccgggccac ccagctgccc aggtccagca gggtgaagtg ggatggcaat cacgcagctc      420

gttcatgtca tctcaagttt aagactcgac ctcgttttgt ctgcggctgg gtgaatgagc      480

acccaatgca cctcttgctg gagctgggcc cccttgggag ctagctggca gctgcaggcc      540

tagcacgctg tgggagggat gacttacact ttgggaaagt aagaaaaaaa ataaagat        598


<210>  13
<211>  832
<212>  DNA
<213>  Parachlorella sp.


<220>
<221>  misc_feature
<223>  FBPase Promoter

<400>  13
ttggtgtttt taaggtgctc atccggtgaa gatgaatggg gtggttgagt tagtgcaggc       60

acgctggtgt gcagtaccca gtgtatggat ggaagtttgg gggcctgata tgtgtgctcc      120

cgcctttttg gtcgtttctt gcttcaaacc agtggcggat gccagttttt cgtccacagg      180

ccaattcgtt attttgcagt tctatttaat tggaaaggtt caaatgtatc atttgtgtaa      240

tatcacatga catctctgtc acgaagcaga atgaaattaa ctaatgatca ttattcgcac      300

atatgtactc caccataagt accatggtgt tgtgacgttg gaatcagcag caaggtcaaa      360

tcacatgtct tttcctcttt caccaacaca ggtaggcaca ccttcaaact gttctgatat      420

atactgagga cgtacattaa gtcgttaata cacgacgcga aatgatcaga ggcagttcat      480

gctattgaca gtctgaccac agtctgatgc acaccagaac taccaaaatg gcatctcgac      540

ggggtcacac ggggtagcga gcaatgcaaa ctcgtaccgt caataaagct tgcaagcatt      600

ccagcaccga atcttgacgg gcgggtaccc ttcgtcagga aacaaccgtc attcaatatc      660

ggctgtttca ttctggcgtt cccgccaagg accctaagcc ctaatcacag ctttatcccc      720

gttcgcaaag cctgcatcag ccgttgagta tttgtgttca gcccactgac aactaagtcc      780

tccacaactt ctgcttgatt ctgtttggtt gctctgaaac acatacacaa cc              832


<210>  14
<211>  703
<212>  DNA
<213>  Parachlorella sp.


<220>
<221>  misc_feature
<223>  FBPase Terminator

<400>  14
gcgctctgtg actaatccgt tgacctattt agtcattgca tgattagtca gtgcagcagg       60

ctgtgtccat tgtcgcctcc tgactgccag agcctttgtt tggtctttgc cttaacattg      120

tacacccttt gtcgtgcttt tcctgagaca gcatttaatt ttgttgggcg tggagcctgt      180

ggcttacgtg tcctctggtc cgtagttgct cggtgcaaca cgggggtatg ggtggcagag      240

gggctcagtc ccgctcttac ccttgcctgc tgctggccct gatcaatttg agatttattt      300

tcctcttttc gacactgagc ttgcatgact gctgagatgg cggtggtagt tgtctgcatg      360

taatgtgtgg cgttttttgg aagtggccct gccctgccac attggtgcta ccacaccttc      420

tttgatggct taatgggcag accctcctgc aggatcgccg aaacctctgc accgccagtc      480

tgtgtaatta aaattcgaga cgctgttaag tggaacactt tgctacagct gaaatgcgaa      540

gcgaatgcgc agtgatgctc aacttttgca actatgctga gctaggcgac tggtgtttaa      600

ttgcacctac tccctactac ggcctcagct catcacagca gataagaagt aagagcacaa      660

tgataatgcg cttaacaacc ccttagcact gcagcatctg tgg                        703


<210>  15
<211>  642
<212>  DNA
<213>  Parachlorella sp.


<220>
<221>  misc_feature
<223>  EF2 Promoter

<400>  15
gaggactgac gattttgcac catgttagcc aagataagga tgttatgttt gcccagctcg       60

tgaagatttt tgatttccat gcacgtgaag caaagaatat gaataaactt gggctacagt      120

tgtcagcaca aacgcataac tttgctggta cacatcgttc gtagaaaaga tataaattat      180

tatagagcaa ttgtatttgg tgaaattttg tgtgtcttgt cgctgatgga actatcatca      240

cacgcgatgc ggagtgtaga aaaacatcta tagattcgcg ctgaacctcc tcccaccttc      300

caatagctga tgagataaca gcatttgtga ttgcgacgga ttaaagtgtt gagtataaaa      360

aagttagagc atttttcatg cgatcaagtc tcccactcga gaactattac taactttact      420

caggcatgca ataatgtcat aatgttatat tgtcggcgca tctcccccgt ggccttgata      480

ggcaacgctg cgaccacaat ctctccttct ccctgctgtt cataacagcg attcattttc      540

aattcaagtg tgcagtccct gcgactcaca ctaattgcgg ctacctcgag ttgctgcatt      600

acctagcccc tgttggctgc tgtgtattga aactcagcaa cc                         642


<210>  16
<211>  668
<212>  DNA
<213>  Parachlorella sp.


<220>
<221>  misc_feature
<223>  EF2 Terminator

<400>  16
gctacttgtt ggagacgtgg cggctgccag agtcaacagc agtgacagca gcattgctgg       60

gagacttgtg cgctattgcc gcccgacagt tgcagcagtg ggctacagcg cagatgctgt      120

gctacacggt ttatagagtt cctttttgat tttgtggaag agcgcttttg ctgtcagctg      180

gcctgaagta cggggagcta tgggagtgca ccagggcgac ggtgcatcac ctatggagca      240

gccatgtaga gtagcattgc tgcatttacc catgggcagc tgcgttgtgg atgaattcat      300

tttccttgaa tgccttgccc tggttataag taataaaatt caattggcca taggtcgagg      360

aaagaagttg cacgtaccaa acgcagcttt tttggcaaat tgcctgccct gagcactgtg      420

tgtttttcgc tctggagacc gaggccatca gaacttttac cgcagctggc aatagcaggc      480

gatttcaatg ctctggcagt gttgctcgag gataaggtac aatctactgc aaagaaacgg      540

tcgtgcccaa tatgtacacg cgccctaccc tcactgcaac gcacgatagg cagagtatat      600

caaccactca ctggtatact gttgttcaca ggcaacatat catttcaacg tatgcaaagc      660

cattgaac                                                               668


<210>  17
<211>  588
<212>  DNA
<213>  Parachlorella sp.


<220>
<221>  misc_feature
<223>  RPS17 Promoter

<400>  17
caacacctag ttggtaaata ccgttgctga tattgctctg taccagtaaa agagggctgc       60

gatgagcgtt tttagtgcac ttcttcaaca cggaatattt ttcacaaatt ggtatgagaa      120

ccaattttgc aaaatgttcg ccctgtaaag tatcgctctg ggacgatcag cttgacgtaa      180

ttgtaggcga aaagggcgtt caaagtgcag ctttatgtat gaacgtcata aaatataaag      240

catagcacaa tcactgatag aaaatatttg tgcgcattaa aactctcact tctgttgcgg      300

atacaacgac ggaaatgaga agcttgtgta agaagcaatt caagttttca ttttgtcatc      360

taaggtgtga tcctccgata ttcattaccg aatgctgatc tgagttggaa agatggcaat      420

atttagctgt gcacactttg acctccaggc cttggcggga atttagtatt ctagctttcc      480

tattggaacg ataggccagc caagtctcca gcttgtatac gctacaccag cagacatgct      540

ctcaatttag ctgacagtgt cttcatattt gtattatctg ttgtgtct                   588


<210>  18
<211>  455
<212>  DNA
<213>  Parachlorella sp.


<220>
<221>  misc_feature
<223>  RSPS17 Terminator

<400>  18
ggtgcgaata gtgcttcagt aaaaaagtag caacttggtg caatatcgtc agggtcgtgt       60

ggtctgctcg ccagcaagtt ttttggcaca ggagagcgct ttttccgagt accgccaaag      120

ttcaagcatg tgctgtgatt cgctgttgcc tcttatgata attgctcaaa gtttccaagc      180

attctatgtc caccctgcac cactaagttg tatggtgctt attctgcagg ggatgattca      240

tggtgcctaa aaattttgtg ctgctgtcgc gtctgttttc tgtcgcagtt tagtgaatgt      300

aactccaaat accaaacttt tcatcacaat catattgatg cctttgtaag tgaattacag      360

cgttttttgc cataaaaaga agtaccgtga cattggggtc gtcataacaa gaagctttat      420

gaacaagcag cttgatctac gagacttata cataa                                 455


<210>  19
<211>  707
<212>  DNA
<213>  Parachlorella sp.


<220>
<221>  misc_feature
<223>  mitoATPSD Promoter

<400>  19
cctgccagcc tcctgtgctt ttcagcctta tatcacacag ggcaatgcga gcaggtgcag       60

tgggtgcaca cgatcatgca cttgttatgc gtgcacaagg aaatacaata acgggatgag      120

taaattcaga aagcggattc tgagtgtgga tcatctagat taattaggca tgctgcagtt      180

aatggattta tcaaaatgca gctagagggt agctgtgatt gggatgcaca gatgttgatg      240

acagtttatt gaagaaaatg gaaagcgtgc acatgaagat gatgaatgcg agtcaattgc      300

agatgatttg tctgtgtcag tctgtgatgc ttcttatcat ggtgtgcgct tctcatccag      360

ctgaaaatac atcagaatgt aatacttgta caatgcaagc gcaaacattc agcaacagtc      420

acggaaaggc aattgaatgg gcgatatctt cgttttggta aaatttgcat caaatccgct      480

ttggttatcc tcgatgaacg ccttgaagca tttaatccaa ctgtgctgta ctacatggga      540

acttgttgac ccagaggttg cttcccgcca ttcaaggccc ctaacaaatg ttaccaaacc      600

ctcactcttc tatccttcga ttggtccgca aattaccaag gaatcgaggt gtttggtaac      660

attcttagct ttactgctct aggttttgtg taggagactt aggcaac                    707


<210>  20
<211>  519
<212>  DNA
<213>  Parachlorella sp.


<220>
<221>  misc_feature
<223>  mitoATPSD Terminator

<400>  20
taggttcagt tgcgagaaca aaatacagtc cctgaacatt ttcggaacgg cgttttatgc       60

ggctttgttg ttccctagtg catgtggggg tttagaagac taggttttgt cactatcatt      120

gtgggcttga tgcccaccca tgcagcagac ctacaggact gcaggtgggt gaacctgttt      180

ctagaagttt tctggtcaat tgtaatgcca actagatgaa gagaacagaa gagttttctt      240

cattacatct gatggattat gctgaccatg cagccacacc aatgtggttt ctatgctgct      300

ttcagtcatg attagcattg caagcacgta ttctattcaa cacccaacca acgagatgaa      360

cataccaaat gtcaattgct tgcaaagcaa tcactcctgc acgagtgcca atttatcatg      420

atggcagtat ttatgcatgt acctttcaac aatatttttt ttcgcatacg aaaaatcgaa      480

cggaaatcta ccttgggcct atgtcaagtg acaggttcg                             519


<210>  21
<211>  874
<212>  DNA
<213>  Parachlorella sp.


<220>
<221>  misc_feature
<223>  RBCS1 Promoter

<400>  21
tggtcagaac ttcactcggc gttctgtctg gaaagggcta tgcagcgctc aattcatttt       60

tatcgactta aacgcaaata cgttcatcca gcagcaccag actcacgaga cgagtggtta      120

cggtgttggc aaaagtgcga ctgatataca aggagaatca gatgtaatat tcgtatctgg      180

attttatctg gtaactgtgc gcgagggtga gttgcgagga ttactgttcc ctgagtgtaa      240

caaattgata tttcccgttc tgcaaaccaa aaagggaata ctttggcaac tatttcacag      300

cctttcccgg gagatttcca tctgcaaatg aaaccggcat caccgcttgc gaagtacaga      360

actcgctgcg tgtgggcgac agcttcgaac ggccgcacta ttttggcaga gggagataag      420

ttgactgcga cacgtaacag gtgtacacat accaaattta acaaagaaaa cgttgtgaag      480

gacagtcatt taaggtactg aagcaggaac ggataaactt cagtgaagat aataatgtag      540

aactatctgt gcgcaatgct cttctactgg gtttgttgaa tatagaacga acaagaaaag      600

agaaaaaagc aatcaggcgt agttcaatgc tgcaaaggtg tctcacaagg tagtccgacg      660

ccacccaagc aagatcgttg atgttggtgt tcatttgaaa gcgacccctc cccggggtcc      720

cagggaaata ctgttgcgcg ccaaatgtgg tagttatctt tctttctttt tgacacagag      780

ataatatgca tcgaaagcac cacaaaagtc cttctcggtc ttgcctgcaa gcttgtcctt      840

tgcgttgctc ctctcaaaaa aacttttgca cgca                                  874


<210>  22
<211>  322
<212>  DNA
<213>  Parachlorella sp.


<220>
<221>  misc_feature
<223>  RBCS1 Terminator

<400>  22
atgagtgccc atatgtaatt cgtagttgat gccatctgaa ggctattcga gaggaaaatc       60

cattttgatg tctttggttt gtaatattac tggtactgct tttcaggtag tatgtgctct      120

attgggacca ggtttttgtg atcgctgaat tgcgctgctc agcctggcgt gatggttttc      180

atctgctgtt ccttgtggat tgctattttg ttgccatgag ttccttctag tgcacaagcc      240

cactgtaaag cgctcgcccc agtcaattat cgatcaatgt caagtgcagt tatcacatga      300

gttaattatt ggttcgtcaa ag                                               322


<210>  23
<211>  874
<212>  DNA
<213>  Parachlorella sp.


<220>
<221>  misc_feature
<223>  RBCS2 Promoter

<400>  23
gaaagcttag cgatacagta ttttcttgtg aagcagaaga agggcaagtg aggtcgttgc       60

ataatgttgt acaccgttgc aattgcatag taaatcagtt tctcaacaga gacatcagtt      120

gggcaattcc atgaaatgca tgttgcaggc caatcatttg cactgtacaa tttgggttgc      180

tcgagaattt gagagtgtac atcagctcga ctgaaagccg ctcaattttg tgcagagaag      240

caatcatcag gtggtaaata ttgaagtgtc gttctctgca agccccattg aaattacaca      300

ttgacgctgc gacaaagtca ggacaacgtc cgatttgtga tccttgacga ataactaaga      360

ctatttatca agcacgcagg tctgcacttg agcgtaggca acaaccgctc catactgggt      420

agcaggaaac ggcccacagc ggaagcaacc cccgtggcat tctagtatta ttgaatatgc      480

agcctgtaac aagacaaagt aggagccaca tgcttggatg taaacacatg catgtttgtt      540

aaataagcaa gcttaagttg ccaagcactc tgcttgacag cttgaaatta agatgcccac      600

caagtgcaac aatctgcgtg ttctttgagg cagtgtcctg atgtattgca agtaccacaa      660

tgggttgatg cattatcgtg gcactcttgc aagcgcatat cagcgtgcat tgggtgcgct      720

aactggccat cggcgcgggt aattttacgc gcgtcacgtg ctgtggatat aggagtattc      780

tgactttctt caaacagtgc ttatcagaac gctttgcggc tgtgtgcatc agttcacacc      840

ttcttggcaa aattctctgc acattgttga cgca                                  874


<210>  24
<211>  510
<212>  DNA
<213>  Parachlorella sp.


<220>
<221>  misc_feature
<223>  RBCS2 Terminator

<400>  24
actttcacac ttcacagaag ctcttacttc aatgccaggg agcatagctc gagaagataa       60

attgatgtgg ctgtgactgt cgggcagttg aactgtgaat gtacgtgctg tagtttttgt      120

tctcattcgt gggtgggtga ttgtctgcct acttttcctg ttcccttcat tcccatttct      180

tgcctgtact cgtggcgcag ctcaccaatt aatttgagca tcgctggtca tctaccacct      240

gattgcgaac atttgctcaa ttctactgtt ccctggtgcg tggtctgcaa taattagctg      300

gggcgagcgc ttgcagttcc actgactgtg ggttatggtt tgttttaaac aaattttgtt      360

ttactaattt cttgtagtgc gttggtcgtt gccatttgca gcattgaaaa gtcaggaagc      420

agacaatcta cccacccatc caatgtaggc gaaaatccag cagtagtccg ctgtatacaa      480

aaaccgatca ctcagcagcc cccccgatga                                       510


<210>  25
<211>  156
<212>  DNA
<213>  Parachlorella sp.


<220>
<221>  misc_feature
<223>  RPS4 Intron 1

<400>  25
gtgagttctg agaagctgat tgttgtttaa cttctttgaa agctttatcg aagattctgc       60

aagcgatgaa cattgcttgt caagaccgag agctgcatgc ccacttgaca tccagctttg      120

aacggctctt catgtttgat ttgtttctga ttgtag                                156


<210>  26
<211>  292
<212>  DNA
<213>  Parachlorella sp.


<220>
<221>  misc_feature
<223>  RPS4 Intron 2

<400>  26
gtgagtgcag cgtcagctgt ggcagttgtt ggctttcgtc tcagtcagta gtttgctggg       60

attgattatg gagggcacag ttgcaatttt gagttgcacg ttgcgacaag cgtgttgaca      120

aagcgtggtc aagccggcca gtcttgccgg tggcgggtgg cttggtctaa cttccgctct      180

acagcaatcg ttttgttcat ggttacgggg ctggcgtgcc agaaagtcct ggtcagccac      240

cctcgcttca aagccgtagc ccaacaactt tgcgaatatg ttcgatttgc ag              292


<210>  27
<211>  1322
<212>  DNA
<213>  Parachlorella sp.


<220>
<221>  misc_feature
<223>  RPS4 Intron 3

<400>  27
gtacagctct gcgtgcaaca ggttgcaaga tgcagcgcag gtcttccctg gtcaaacgat       60

gtatgcagag ttgagaggca cttgagctgg gtgaatggcg tgggctcgta ggtagtgtgc      120

agggcaggaa gggcagccaa ttttggagtt gtggtccggt gtcgttgctt cgagccttat      180

taggactctt gctcatcaaa gcgttagttg tgaataagtt gatctgaaag gatgttatgt      240

acagcaagca gcagcagtta agagtctggg gagtagctgc acagggcgag gtgtcaagat      300

gggaagggtc ctgcctcctt atgtgttttt ccctgtaggg gaggaagcct cttatgggca      360

atggttgggc atattttcca gccagccctt ctttctatag gggccagggt gggcccagct      420

cgtcttggct tccaccacca ggagagtgag ggcattgaag ggccataaat agtcctccca      480

tctacgtgca ccagagggtg tcgtctaggc tgtgcatgcc acgaggggaa ggagccaaga      540

atgagtgtat gggttgtttt catgtttagg ctgggataaa actgttttca attgcgcctg      600

ccgggtgaaa accacagcag catcagcaag cttggagaag gccagcccgc ccagcacagg      660

ctcacgttcc cactcaggcg gtcagtcggg cgggggtgtg agtcaggcag gcgagggtgt      720

ctgtgcctga catcagcacc tctgcttagc cactgcagcc cctggagcag ggtagggcgt      780

catttgcagc aatcacctgc tgcctcacac gtcgcagctt ggaatttcaa cgaccatcag      840

cgctggggtt gttgagggat catagcagat tttggtgcag cctggttgtc atgctctttg      900

tggaatggcc tctatgttcg agcaattcgt tggatgttga ggtgcttggg gacagagagt      960

cgaatgatgg gccagggtca aacatgcgag cgtttggctg agtcagcggt ttttgctggt     1020

cactttttct tttgtttctt atttaggttt gatggatgtg ttttgtgctg ctgccctgaa     1080

gctgcagcag cgtgtctgcc ctgcgctact gcgggcacca aggctatgtg ctggtgcact     1140

cggctgcgct gcacctgtgc acctcgcact ccgtccagcc tccatgcagc acacgtactc     1200

acggtgtcct cctgacctgt cgtacgctat tccaaacttg ctcttttgct gccgctgctc     1260

tcgtacacaa ttgctgttga ttatcgatat ctaatcgagc gcctgctgac tgaactccgc     1320

ag                                                                    1322


<210>  28
<211>  326
<212>  DNA
<213>  Parachlorella sp.


<220>
<221>  misc_feature
<223>  RPS4 Intron 4

<400>  28
gtgggtgggc tctgaaggag gaggagggag cgggtgatta aacagggcct gcatgaagag       60

gagcaggggc tgcgtggaca gcagggggaa ggtgcagaag ggagggtcaa gcggggttca      120

ggtggctgtg ggtttctgca cgagcagtga aagaagctgt atccttccac ctgcttccac      180

tggcgaaagg ttgaaaacag gatgtcgcag ctggaaagat gttgcgctgt caagtgcaag      240

ccatggttga gggtatgcct gtgtgcatgt gcttcttaaa gttactcctg ttctatggtt      300

ctgggtgctt gttgtttgtg gtgcag                                           326


<210>  29
<211>  196
<212>  DNA
<213>  Parachlorella sp.


<220>
<221>  misc_feature
<223>  RPS4 Intron 5

<400>  29
gtgagggggc atgtaagcaa tggcaggcaa ttcaagaacg aatcattgct gcaaatgctg       60

ggatggtatg cagctgaggt atctattgcc ttgtattttg tctcgcattg catcggtggt      120

gcgttctgtg gcctgaggca cagttcttgc tgtttgataa gggttcgact gagttgtcgt      180

gtgtgctgtg ctgcag                                                      196


<210>  30
<211>  2667
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic


<220>
<221>  misc_feature
<223>  Intronylated, codon-optimized BleR gene

<400>  30
atggccaaac tgacatccgc tgttcctgtg ttgacagcaa gagatgttgc aggtgcagtg       60

gagttttgtg agttctgaga agctgattgt tgtttaactt ctttgaaagc tttatcgaag      120

attctgcaag cgatgaacat tgcttgtcaa gaccgagagc tgcatgccca cttgacatcc      180

agctttgaac ggctcttcat gtttgatttg tttctgattg tagggacaga tagactgggg      240

tttagcaggg actttgtgga ggacgatttt gcaggagtgg tgagggatga tgtgacactg      300

tttatctcag cagtgcagga tcaagtgagt gcagcgtcag ctgtggcagt tgttggcttt      360

cgtctcagtc agtagtttgc tgggattgat tatggagggc acagttgcaa ttttgagttg      420

cacgttgcga caagcgtgtt gacaaagcgt ggtcaagccg gccagtcttg ccggtggcgg      480

gtggcttggt ctaacttccg ctctacagca atcgttttgt tcatggttac ggggctggcg      540

tgccagaaag tcctggtcag ccaccctcgc ttcaaagccg tagcccaaca actttgcgaa      600

tatgttcgat ttgcaggtgg tgcccgataa tacactggca tgggtttggg tgagaggtac      660

agctctgcgt gcaacaggtt gcaagatgca gcgcaggtct tccctggtca aacgatgtat      720

gcagagttga gaggcacttg agctgggtga atggcgtggg ctcgtaggta gtgtgcaggg      780

caggaagggc agccaatttt ggagttgtgg tccggtgtcg ttgcttcgag ccttattagg      840

actcttgctc atcaaagcgt tagttgtgaa taagttgatc tgaaaggatg ttatgtacag      900

caagcagcag cagttaagag tctggggagt agctgcacag ggcgaggtgt caagatggga      960

agggtcctgc ctccttatgt gtttttccct gtaggggagg aagcctctta tgggcaatgg     1020

ttgggcatat tttccagcca gcccttcttt ctataggggc cagggtgggc ccagctcgtc     1080

ttggcttcca ccaccaggag agtgagggca ttgaagggcc ataaatagtc ctcccatcta     1140

cgtgcaccag agggtgtcgt ctaggctgtg catgccacga ggggaaggag ccaagaatga     1200

gtgtatgggt tgttttcatg tttaggctgg gataaaactg ttttcaattg cgcctgccgg     1260

gtgaaaacca cagcagcatc agcaagcttg gagaaggcca gcccgcccag cacaggctca     1320

cgttcccact caggcggtca gtcgggcggg ggtgtgagtc aggcaggcga gggtgtctgt     1380

gcctgacatc agcacctctg cttagccact gcagcccctg gagcagggta gggcgtcatt     1440

tgcagcaatc acctgctgcc tcacacgtcg cagcttggaa tttcaacgac catcagcgct     1500

ggggttgttg agggatcata gcagattttg gtgcagcctg gttgtcatgc tctttgtgga     1560

atggcctcta tgttcgagca attcgttgga tgttgaggtg cttggggaca gagagtcgaa     1620

tgatgggcca gggtcaaaca tgcgagcgtt tggctgagtc agcggttttt gctggtcact     1680

ttttcttttg tttcttattt aggtttgatg gatgtgtttt gtgctgctgc cctgaagctg     1740

cagcagcgtg tctgccctgc gctactgcgg gcaccaaggc tatgtgctgg tgcactcggc     1800

tgcgctgcac ctgtgcacct cgcactccgt ccagcctcca tgcagcacac gtactcacgg     1860

tgtcctcctg acctgtcgta cgctattcca aacttgctct tttgctgccg ctgctctcgt     1920

acacaattgc tgttgattat cgatatctaa tcgagcgcct gctgactgaa ctccgcaggt     1980

ttggatgaac tgtatgcaga gtggtctgaa gtggtgagca ccaactttag gtgggtgggc     2040

tctgaaggag gaggagggag cgggtgatta aacagggcct gcatgaagag gagcaggggc     2100

tgcatggaca gcagggggaa ggtgcagaag ggagggtcaa gcggggttca ggtggctgtg     2160

ggtttctgca cgagcagtga aagaagctgt atccttccac ctgctttcac tggcgaaagg     2220

ttgaaaacag gatgtcgcag ctggaaagat gttgcgctgt caagtgcaag ccatggttga     2280

gggtatgcct gtgtgcatgt gcttcttaaa gttactcctg ttctatggtt ctgggtgctt     2340

gttgtttgtg gtgcagggat gcaagcggac ctgcaatgac agagattgga gaacaacctt     2400

ggggaaggga gtttgcattg agagatcctg caggtgaggg ggcatgtaag caatggcagg     2460

caattcaaga acgaatcatt gctgcaaatg ctgggatggt atgcagctga ggtatctatt     2520

gccttgtatt ttgtctcgca ttgcatcggt ggtgcgttct gtggcctgag gcacagttct     2580

tgctgtttga taagggttcg actgagttgt cgtgtgtgct gtgctgcagg caattgcgtg     2640

cactttgttg cagaagaaca ggactga                                         2667


<210>  31
<211>  71
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic


<220>
<221>  misc_feature
<223>  plasmid cloning site

<400>  31
cctgcaggca gttggtacgg catattatgg tttaaacatc tatcctccag atcaccaggg       60

ccagtgaggc c                                                            71


<210>  32
<211>  399
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic


<220>
<221>  misc_feature
<223>  Codon-optimized Bsd gene

<400>  32
atggcaaaac ctctctccca ggaagagtct accctgattg aaagggcaac agcaaccatc       60

aacagcattc ccattagcga ggactactct gttgcatctg cagcattgag ctctgatgga      120

aggatcttta caggagtgaa cgtgtaccac tttacaggag gaccttgtgc agaattggtg      180

gtgttaggta cagcagctgc agcagcagca ggaaatttga catgcattgt ggcaataggg      240

aacgagaata gggggatttt gtcaccttgc ggaagatgta gacaggtgtt gttggatctg      300

catcccggga ttaaggcaat cgtgaaggat tcagatgggc agcctacagc agtgggaatt      360

agagaactgc tgccttctgg gtatgtgtgg gaaggataa                             399


<210>  33
<211>  2691
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic


<220>
<221>  misc_feature
<223>  Bsd gene, intronylated

<400>  33
atggcaaaac ctctctccca ggaagagtct accctgattg aaagggcaac agcaaccatc       60

aacagcattc ccattagcga ggactactct gttgtgagtt ctgagaagct gattgttgtt      120

taacttcttt gaaagcttta tcgaagattc tgcaagcgat gaacattgct tgtcaagacc      180

gagagctgca tgcccacttg acatccagct ttgaacggct cttcatgttt gatttgtttc      240

tgattgtagg catctgcagc attgagctct gatggaagga tctttacagg agtgagtgca      300

gcgtcagctg tggcagttgt tggctttcgt ctcagtcagt agtttgctgg gattgattat      360

ggagggcaca gttgcaattt tgagttgcac gttgcgacaa gcgtgttgac aaagcgtggt      420

caagccggcc agtcttgccg gtggcgggtg gcttggtcta acttccgctc tacggcaatc      480

gttttgttca tggttacggg gctggcgtgc cagaaagtcc tggtcagcca ccctcgcttc      540

aaagccgtag cccaacaact ttgcgaatat gttcgatttg caggtgaacg tgtaccactt      600

tacaggagga ccttgtgcag aattggtggt gttaggtaca gctctgcgtg caacaggttg      660

caagatgcag cgcaggtctt ccctggtcaa acgatgtatg cagagttgag aggcacttga      720

gctgggtgaa tggcgtgggc tcgtaggtag tgtgcagggc aggaagggca gccaattttg      780

gagttgtggt ccggtgtcgt tgcttcgagc cttattagga ctcttgctca tcaaagcgtt      840

agttgtgaat aagttgatct gaaaggatgt tatgtacagc aagcagcagc agttaagagt      900

ctggggagta gctgcacagg gcgaggtgtc aagatgggaa gggtcctgcc tccttatgtg      960

tttttccctg taggggagga agcctcttat gggcaatggt tgggcatatt ttccagccag     1020

cccttctttc tataggggcc agggtgggcc cagctcgtct tggcttccac caccaggaga     1080

gtgagggcat tgaagggcca taaatagtcc tcccatctac gtgcaccaga gggtgtcgtc     1140

taggctgtgc atgccacgag gggaaggagc caagaatgag tgtatgggtt gttttcatgt     1200

ttaggctggg ataaaactgt tttcaattgc gcctgccggg tgaaaaccac agcagcatca     1260

gcaagcttgg agaaggccag cccgcccagc acaggctcac gttcccactc aggcggtcag     1320

tcgggcgggg gtgtgagtca ggcaggcgag ggtgtctgtg cctgacatca gcacctctgc     1380

ttagccactg cagcccctgg agcagggtag ggcgtcattt gcagcaatca cctgctgcct     1440

cacacgtcgc agcttggaat ttcaacgacc atcagcgctg gggttgttga gggatcatag     1500

cagattttgg tgcagcctgg ttgtcatgct ctttgtggaa tggcctctat gttcgagcaa     1560

ttcgttggat gttgaggtgc ttggggacag agagtcgaat gatgggccag ggtcaaacat     1620

gcgagcgttt ggctgagtca gcggtttttg ctggtcactt tttcttttgt ttcttattta     1680

ggtttgatgg atgtgttttg tgctgctgcc ctgaagctgc agcagcgtgt ctgccctgcg     1740

ctactgcggg caccaaggct atgtgctggt gcactcggct gcgctgcacc tgtgcacctc     1800

gcactccgtc cagcctccat gcagcacacg tactcacggt gtcctcctga cctgtcgtac     1860

gctattccaa acttgctctt ttgctgccgc tgctctcgta cacaattgct gttgattatc     1920

gatatctaat cgagcgcctg ctgactgaac tccgcaggta cagcagctgc agcagcagca     1980

ggaaatttga catgcattgt ggcaataggt gggtgggctc tgaaggagga ggagggagcg     2040

ggtgattaaa cagggcctgc atgaagagga gcaggggctg cgtggacagc agggggaagg     2100

tgcagaaggg agggtcaagc ggggttcagg tggctgtggg tttctgcacg agcagtgaaa     2160

gaagctgtat ccttccacct gcttccactg gcgaaaggtt gaaaacagga tgtcgcagct     2220

ggaaagatgt tgcgctgtca agtgcaagcc atggttgagg gtatgcctgt gtgcatgtgc     2280

ttcttaaagt tactcctgtt ctatggttct gggtgcttgt tgtttgtggt gcagggaacg     2340

agaatagggg gattttgtca ccttgcggaa gatgtagaca ggtgttgttg gatctgcatc     2400

ccgggattaa ggtgaggggg catgtaagca atggcaggca attcaagaac gaatcattgc     2460

tgcaaatgct gggatggtat gcagctgagg tatctattgc cttgtatttt gtctcgcatt     2520

gcatcggtgg tgcgttctgt ggcctgaggc acagttcttg ctgtttgata agggttcgac     2580

tgagttgtcg tgtgtgctgt gctgcaggca atcgtgaagg attcagatgg gcagcctaca     2640

gcagtgggaa ttagagaact gctgccttct gggtatgtgt gggaaggata a              2691


<210>  34
<211>  1385
<212>  PRT
<213>  Artificial Sequence

<220>
<223>  Synthetic


<220>
<221>  misc_feature
<223>  Encodes S. pyogenes Cas9, codon-optimized for Parachlorella, with
       NLS and C terminal FLAG tag

<400>  34

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Asp Lys Lys Tyr Ser Ile 
1               5                   10                  15      


Gly Leu Asp Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp 
            20                  25                  30          


Glu Tyr Lys Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asp 
        35                  40                  45              


Arg His Ser Ile Lys Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser 
    50                  55                  60                  


Gly Glu Thr Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg 
65                  70                  75                  80  


Tyr Thr Arg Arg Lys Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ser 
                85                  90                  95      


Asn Glu Met Ala Lys Val Asp Asp Ser Phe Phe His Arg Leu Glu Glu 
            100                 105                 110         


Ser Phe Leu Val Glu Glu Asp Lys Lys His Glu Arg His Pro Ile Phe 
        115                 120                 125             


Gly Asn Ile Val Asp Glu Val Ala Tyr His Glu Lys Tyr Pro Thr Ile 
    130                 135                 140                 


Tyr His Leu Arg Lys Lys Leu Val Asp Ser Thr Asp Lys Ala Asp Leu 
145                 150                 155                 160 


Arg Leu Ile Tyr Leu Ala Leu Ala His Met Ile Lys Phe Arg Gly His 
                165                 170                 175     


Phe Leu Ile Glu Gly Asp Leu Asn Pro Asp Asn Ser Asp Val Asp Lys 
            180                 185                 190         


Leu Phe Ile Gln Leu Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn 
        195                 200                 205             


Pro Ile Asn Ala Ser Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg 
    210                 215                 220                 


Leu Ser Lys Ser Arg Arg Leu Glu Asn Leu Ile Ala Gln Leu Pro Gly 
225                 230                 235                 240 


Glu Lys Lys Asn Gly Leu Phe Gly Asn Leu Ile Ala Leu Ser Leu Gly 
                245                 250                 255     


Leu Thr Pro Asn Phe Lys Ser Asn Phe Asp Leu Ala Glu Asp Ala Lys 
            260                 265                 270         


Leu Gln Leu Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Asn Leu Leu 
        275                 280                 285             


Ala Gln Ile Gly Asp Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn 
    290                 295                 300                 


Leu Ser Asp Ala Ile Leu Leu Ser Asp Ile Leu Arg Val Asn Thr Glu 
305                 310                 315                 320 


Ile Thr Lys Ala Pro Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu 
                325                 330                 335     


His His Gln Asp Leu Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu 
            340                 345                 350         


Pro Glu Lys Tyr Lys Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr 
        355                 360                 365             


Ala Gly Tyr Ile Asp Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe 
    370                 375                 380                 


Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val 
385                 390                 395                 400 


Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn 
                405                 410                 415     


Gly Ser Ile Pro His Gln Ile His Leu Gly Glu Leu His Ala Ile Leu 
            420                 425                 430         


Arg Arg Gln Glu Asp Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys 
        435                 440                 445             


Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu 
    450                 455                 460                 


Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu 
465                 470                 475                 480 


Thr Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser 
                485                 490                 495     


Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Lys Asn Leu Pro 
            500                 505                 510         


Asn Glu Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr 
        515                 520                 525             


Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg 
    530                 535                 540                 


Lys Pro Ala Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu 
545                 550                 555                 560 


Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp 
                565                 570                 575     


Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ser Gly Val 
            580                 585                 590         


Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys 
        595                 600                 605             


Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile 
    610                 615                 620                 


Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met 
625                 630                 635                 640 


Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val 
                645                 650                 655     


Met Lys Gln Leu Lys Arg Arg Arg Tyr Thr Gly Trp Gly Arg Leu Ser 
            660                 665                 670         


Arg Lys Leu Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile 
        675                 680                 685             


Leu Asp Phe Leu Lys Ser Asp Gly Phe Ala Asn Arg Asn Phe Met Gln 
    690                 695                 700                 


Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu Asp Ile Gln Lys Ala 
705                 710                 715                 720 


Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu His Ile Ala Asn Leu 
                725                 730                 735     


Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Val 
            740                 745                 750         


Val Asp Glu Leu Val Lys Val Met Gly Arg His Lys Pro Glu Asn Ile 
        755                 760                 765             


Val Ile Glu Met Ala Arg Glu Asn Gln Thr Thr Gln Lys Gly Gln Lys 
    770                 775                 780                 


Asn Ser Arg Glu Arg Met Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu 
785                 790                 795                 800 


Gly Ser Gln Ile Leu Lys Glu His Pro Val Glu Asn Thr Gln Leu Gln 
                805                 810                 815     


Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr 
            820                 825                 830         


Val Asp Gln Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp 
        835                 840                 845             


His Ile Val Pro Gln Ser Phe Leu Lys Asp Asp Ser Ile Asp Asn Lys 
    850                 855                 860                 


Val Leu Thr Arg Ser Asp Lys Asn Arg Gly Lys Ser Asp Asn Val Pro 
865                 870                 875                 880 


Ser Glu Glu Val Val Lys Lys Met Lys Asn Tyr Trp Arg Gln Leu Leu 
                885                 890                 895     


Asn Ala Lys Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr Lys Ala 
            900                 905                 910         


Glu Arg Gly Gly Leu Ser Glu Leu Asp Lys Ala Gly Phe Ile Lys Arg 
        915                 920                 925             


Gln Leu Val Glu Thr Arg Gln Ile Thr Lys His Val Ala Gln Ile Leu 
    930                 935                 940                 


Asp Ser Arg Met Asn Thr Lys Tyr Asp Glu Asn Asp Lys Leu Ile Arg 
945                 950                 955                 960 


Glu Val Lys Val Ile Thr Leu Lys Ser Lys Leu Val Ser Asp Phe Arg 
                965                 970                 975     


Lys Asp Phe Gln Phe Tyr Lys Val Arg Glu Ile Asn Asn Tyr His His 
            980                 985                 990         


Ala His Asp Ala Tyr Leu Asn Ala  Val Val Gly Thr Ala  Leu Ile Lys 
        995                 1000                 1005             


Lys Tyr  Pro Lys Leu Glu Ser  Glu Phe Val Tyr Gly  Asp Tyr Lys 
    1010                 1015                 1020             


Val Tyr  Asp Val Arg Lys Met  Ile Ala Lys Ser Glu  Gln Glu Ile 
    1025                 1030                 1035             


Gly Lys  Ala Thr Ala Lys Tyr  Phe Phe Tyr Ser Asn  Ile Met Asn 
    1040                 1045                 1050             


Phe Phe  Lys Thr Glu Ile Thr  Leu Ala Asn Gly Glu  Ile Arg Lys 
    1055                 1060                 1065             


Arg Pro  Leu Ile Glu Thr Asn  Gly Glu Thr Gly Glu  Ile Val Trp 
    1070                 1075                 1080             


Asp Lys  Gly Arg Asp Phe Ala  Thr Val Arg Lys Val  Leu Ser Met 
    1085                 1090                 1095             


Pro Gln  Val Asn Ile Val Lys  Lys Thr Glu Val Gln  Thr Gly Gly 
    1100                 1105                 1110             


Phe Ser  Lys Glu Ser Ile Leu  Pro Lys Arg Asn Ser  Asp Lys Leu 
    1115                 1120                 1125             


Ile Ala  Arg Lys Lys Asp Trp  Asp Pro Lys Lys Tyr  Gly Gly Phe 
    1130                 1135                 1140             


Asp Ser  Pro Thr Val Ala Tyr  Ser Val Leu Val Val  Ala Lys Val 
    1145                 1150                 1155             


Glu Lys  Gly Lys Ser Lys Lys  Leu Lys Ser Val Lys  Glu Leu Leu 
    1160                 1165                 1170             


Gly Ile  Thr Ile Met Glu Arg  Ser Ser Phe Glu Lys  Asn Pro Ile 
    1175                 1180                 1185             


Asp Phe  Leu Glu Ala Lys Gly  Tyr Lys Glu Val Lys  Lys Asp Leu 
    1190                 1195                 1200             


Ile Ile  Lys Leu Pro Lys Tyr  Ser Leu Phe Glu Leu  Glu Asn Gly 
    1205                 1210                 1215             


Arg Lys  Arg Met Leu Ala Ser  Ala Gly Glu Leu Gln  Lys Gly Asn 
    1220                 1225                 1230             


Glu Leu  Ala Leu Pro Ser Lys  Tyr Val Asn Phe Leu  Tyr Leu Ala 
    1235                 1240                 1245             


Ser His  Tyr Glu Lys Leu Lys  Gly Ser Pro Glu Asp  Asn Glu Gln 
    1250                 1255                 1260             


Lys Gln  Leu Phe Val Glu Gln  His Lys His Tyr Leu  Asp Glu Ile 
    1265                 1270                 1275             


Ile Glu  Gln Ile Ser Glu Phe  Ser Lys Arg Val Ile  Leu Ala Asp 
    1280                 1285                 1290             


Ala Asn  Leu Asp Lys Val Leu  Ser Ala Tyr Asn Lys  His Arg Asp 
    1295                 1300                 1305             


Lys Pro  Ile Arg Glu Gln Ala  Glu Asn Ile Ile His  Leu Phe Thr 
    1310                 1315                 1320             


Leu Thr  Asn Leu Gly Ala Pro  Ala Ala Phe Lys Tyr  Phe Asp Thr 
    1325                 1330                 1335             


Thr Ile  Asp Arg Lys Arg Tyr  Thr Ser Thr Lys Glu  Val Leu Asp 
    1340                 1345                 1350             


Ala Thr  Leu Ile His Gln Ser  Ile Thr Gly Leu Tyr  Glu Thr Arg 
    1355                 1360                 1365             


Ile Asp  Leu Ser Gln Leu Gly  Gly Asp Asp Tyr Lys  Asp Asp Asp 
    1370                 1375                 1380             


Asp Lys  
    1385 


<210>  35
<211>  221
<212>  DNA
<213>  Parachlorella sp.


<220>
<221>  misc_feature
<223>  FBPase Intron 1

<400>  35
gtgagtgaga acagttttca gatcgaatag cacccccccg cctctgcagc agtcgcatac       60

cggctgcagt aatagcttgg ttcaacggcg acctgaacaa gtactgtagt ttctatgcat      120

acgaacttta tcgaatagaa tcacgcttgg gtatcgatca taccttagcg ctcaatttca      180

ttggctgcta cagaccatat tttcctcttc acttgttgca g                          221


<210>  36
<211>  184
<212>  DNA
<213>  Parachlorella sp.


<220>
<221>  misc_feature
<223>  FBPase Intron 2

<400>  36
gtgagaagag tttggctacc aaatctatct tttcatatca catataccgc ctgatattct       60

gaggtggtgg cttttgtctt tttctttcag tatttttctt cgttgggaac ctaccgcgag      120

ggcattcatt gtggcggatc tgtaagtgcg accaggctgt atccaatatt ttttcctatc      180

gcag                                                                   184


<210>  37
<211>  266
<212>  DNA
<213>  Parachlorella sp.


<220>
<221>  misc_feature
<223>  FBPase Intron 3

<400>  37
gtgagaatct ctgcttgtcg aatgtgtcca gttgtgtctt gaatcctggc aagatgttct       60

tttcaccatc cgtcctgcaa aagtgtcaga agtagcatct ctcgatcgcg ttgtcacttc      120

aacgcctccg caactccccc cgttgtgaat cctgtggtca tggctcagct tttcagatct      180

ctacctgcat gttgtttgcc tgtctcagtc ctgcctgcac aaatcatcgc ccttgtttac      240

tccttgcaat cacggattgt gtgcag                                           266


<210>  38
<211>  429
<212>  DNA
<213>  Parachlorella sp.


<220>
<221>  misc_feature
<223>  FBPase Intron 4

<400>  38
gtgagtggag ggctggggtt tgggggtggg gggtggggag ggaggcacgg atggtgtttt       60

ctcatgtcca accgtggttc atgcaaccga acagcagttt cacaagatgg ttccaacagg      120

gtgctccatt tctccctgac aaaacctcgt gcggtccatc tggtatagct gggttagtag      180

ggggttgtgg gctgtccaca gtcagtgcga agcaggctct attgagcgtg tgctagtgtg      240

tgctgtgctg attggcattt tgttgggccg agtgttagga ttagggtaaa tcaccctaat      300

taaccttaca taataggact gtatgcaaat ttgttttcca aaaactctac ccagcgtggt      360

cagactgcat gcactgtgga gcatgcatgg ggctgaccct gttgatcctg ctcattctgc      420

ttcctccag                                                              429


<210>  39
<211>  256
<212>  DNA
<213>  Parachlorella sp.


<220>
<221>  misc_feature
<223>  FBPase Intron 5

<400>  39
gtaggtgcag gaagaagtga atgatgcaca catggtggaa tcgtgataca agcagcagca       60

agtgttggac caagacatgt gcgtgctttg ctgctgccaa gctggcactg caccaggtcg      120

tgcattgatc tgcacatttg atatactgtg agagtcagac gacgtccttt cagagcctgt      180

gtgtgattct ccaggggtta acacgagttt cctttctgcc agtgagtcac cctctcgctg      240

ctcgctcctg gtgcag                                                      256


<210>  40
<211>  338
<212>  DNA
<213>  Parachlorella sp.


<220>
<221>  misc_feature
<223>  FBPase Intron 6

<400>  40
gtgataatta tgcaagaaaa tgaaaaaaat catagaaagg aaggagggga tgttgatgtt       60

tggtgtgtgg cagggtgtgt gttggactga tgcctattgg attgccggtg gcctgcatgt      120

cagctgcttc tcactaaatt atcgctcggg aaatgcgcag tccacaacta ttcccaacat      180

caatgcccaa acagttgcaa gacagctgct gcttgcccca ccctttgatg gtcttctgca      240

cctggaaagg gtgcactgtt tgtcttcttt gagcttcaag ctcgtctccc tgctccttgc      300

ctcacttgcc ttcctcgtct gcccctccct ttctgcag                              338


<210>  41
<211>  259
<212>  DNA
<213>  Parachlorella sp.


<220>
<221>  misc_feature
<223>  FBPase Intron 7

<400>  41
gtgagtgcgg gacatcacat agggagggga gggagggatt gggtggaggg ggagatgctg       60

gtcttttgct gagatgcaag tgtgggtcca cgcagacgtg ggttgactgt ccaacagacc      120

agcagctatc acaagttata ccaccatgca cttatgagaa cttccatcag tttcctttgg      180

catgcacctg aatgccaatt ggtcttgctt tggcctgcac tcatgcttgc actcctgccc      240

ctctccccgc tacatgcag                                                   259


<210>  42
<211>  231
<212>  DNA
<213>  Parachlorella sp.


<220>
<221>  misc_feature
<223>  FBPase Intron 8

<400>  42
gtgagtggca ttaagtagca ggattgaatt tatttcctgg acttggccag ataacatgtg       60

acacaccatg gtagagaagt tgtttgttct ggcgaagaac ccttgacaaa ttgtgtcatc      120

cgtttttgga ctatatatcg tttgaaatgc aaaggcctcc ataaaagatc aggcagtgcg      180

cccaagtttt gagcaggaag ctgacccctg tgcaccatgc tgtgctgtgc a               231


<210>  43
<211>  237
<212>  DNA
<213>  Parachlorella sp.


<220>
<221>  misc_feature
<223>  FBPase Intron 9

<400>  43
gtggggaata ctttttggca agagcgtgtt ttgtggtttc tcgctcacct gccctgacta       60

gaacttctgt tgacaggccc acggtctgta ggccacgtct gtcagaccat gtggaacttt      120

cttctattgc ataacgcgag ctgccccaca tgttaatcct tgttgggtgt ctcgcatcac      180

atcatgtcgc ctttcatgca cttctaacca ttatgtgaac accctcgccc tctgcag         237


<210>  44
<211>  5540
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic


<220>
<221>  misc_feature
<223>  Gene encoding Streptococcus pyogenes Cas9 with FLAG tag, nuclear 
       localization sequences, peptide linker, and FBPase introns 1-5, 
       codon optimized for expression in Parachlorella

<400>  44
atgcccaaga agaagcggaa agtcggggac tacaaggacg acgatgacaa actggagcct       60

ggggagaagc cctataagtg tcctgagtgc gggaagagct tcagccaatc tggagcactg      120

acaaggcacc agaggacaca tacacgcgac aagaagtaca gcatcgggct ggatatcggg      180

accaattctg tgggatgggc cgtgattacc gacgagtata aggtgcccag caagaagttc      240

aaggtgctgg ggaacacaga ccgccacagc attaagaaga acctgatcgg ggcgctgctg      300

tttgattctg gagagacagc agaggcaacc gtgagtgaga acagttttca gatcgaatag      360

cacccccccg cctctgcagc agtcgcatac cggctgcagt aatagcttgg ttcaacggcg      420

acctgaacaa gtactgtagt ttctatgcat acgaacttta tcgaatagaa tcacgcttgg      480

gtatcgatca taccttagcg ctcaatttca ttggctgcta cagaccatat tttcctcttc      540

acttgttgca gcgcctgaaa agaacagcaa gaaggcgcta cacccgccgc aagaatagga      600

tttgctacct gcaagagatc ttcagcaacg agatggccaa ggtggacgac agcttcttcc      660

atagactgga ggagtcgttc ctggtggagg aggataagaa gcacgagagg caccccatct      720

tcggtgagaa gagtttggct accaaatcta tcttttcata tcacatatac cgcctgatat      780

tctgaggtgg tggcttttgt ctttttcttt cagtattttt cttcgttggg aacctaccgc      840

gagggcattc attgtggcgg atctgtaagt gcgaccaggc tgtatccaat attttttcct      900

atcgcaggga acattgtgga tgaggtggcc taccacgaga agtaccccac aatctaccac      960

ctgcgcaaga agctggtgag aatctctgct tgtcgaatgt gtccagttgt gtcttgaatc     1020

ctggcaagat gttcttttca ccatccgtcc tgcaaaagtg tcagaagtag catctctcga     1080

tcgcgttgtc acttcaacgc ctccgcaact ccccccgttg tgaatcctgt ggtcatggct     1140

cagcttttca gatctctacc tgcatgttgt ttgcctgtct cagtcctgcc tgcacaaatc     1200

atcgcccttg tttactcctt gcaatcacgg attgtgtgca ggtggacagc acagataagg     1260

ccgatctgag gctgatctac ctggcattgg cccacatgat caagtttagg gggcacttcc     1320

tcatcgaggg ggatttgaac cccgacaaca gcgatgtgga caagctgttc atccagctgg     1380

tgagtggagg gctggggttt gggggtgggg ggtggggagg gaggcacgga tggtgttttc     1440

tcatgtccaa ccgtggttca tgcaaccgaa cagcagtttc acaagatggt tccaacaggg     1500

tgctccattt ctccctgaca aaacctcgtg cggtccatct ggtatagctg ggttagtagg     1560

gggttgtggg ctgtccacag tcagtgcgaa gcaggctcta ttgagcgtgt gctagtgtgt     1620

gctgtgctga ttggcatttt gttgggccga gtgttaggat tagggtaaat caccctaatt     1680

aaccttacat aataggactg tatgcaaatt tgttttccaa aaactctacc cagcgtggtc     1740

agactgcatg cactgtggag catgcatggg gctgaccctg ttgatcctgc tcattctgct     1800

tcctccaggt gcagacctac aaccagctgt ttgaggagaa ccccatcaac gcatctgggg     1860

ttgacgcaaa ggccattctg tctgcaaggt aggtgcagga agaagtgaat gatgcacaca     1920

tggtggaatc gtgatacaag cagcagcaag tgttggacca agacatgtgc gtgctttgct     1980

gctgccaagc tggcactgca ccaggtcgtg cattgatctg cacatttgat atactgtgag     2040

agtcagacga cgtcctttca gagcctgtgt gtgattctcc aggggttaac acgagtttcc     2100

tttctgccag tgagtcaccc tctcgctgct cgctcctggt gcaggctgag caagtcaagg     2160

agactggaga acctgatcgc ccaattgcct ggagagaaga agaacgggct gttcgggaac     2220

ctgatcgcat tgtctctggg gttgaccccc aacttcaaga gcaacttcga cctggcagag     2280

gacgcaaaac tgcagctgag caaggacacc tacgacgatg atctggacaa cctgctggcc     2340

cagattggag atcagtacgc agacctgttc ctggcagcca agaatctgag cgacgcaatt     2400

ctgctgagcg acattctgcg cgtgaacacc gagatcacca aggcacctct gagcgcaagc     2460

atgatcaaga ggtacgacga gcaccaccaa gacctgacac tgctgaaagc actggtgaga     2520

cagcagctgc ctgagaagta caaggagatc ttcttcgacc agagcaagaa cgggtacgct     2580

gggtacattg atggaggagc aagccaagag gagttctaca agttcatcaa gcccatcctg     2640

gagaagatgg acgggacaga agagttgctg gtgaagctga atcgcgagga tctgctgagg     2700

aagcagagga cattcgacaa tgggagcatc ccacaccaga tccatctggg agagctgcac     2760

gcaattctga ggagacaaga ggacttctac ccgttcctga aggacaatcg cgagaagatc     2820

gagaagatcc tcacgttccg catcccgtac tatgtgggac ctctggcaag ggggaactct     2880

agatttgcct ggatgacccg caagagcgag gagacaatta caccctggaa cttcgaggag     2940

gtggtggata aaggggcatc tgcacagagc ttcatcgaga ggatgaccaa cttcgacaag     3000

aacctgccca acgagaaggt actgcctaag cattcactgc tgtacgagta cttcaccgtg     3060

tacaacgagc tgaccaaggt gaagtacgtg acagagggga tgaggaagcc agcatttctg     3120

agcggagagc aaaagaaggc catcgtggat ctgctgttca agaccaaccg caaggtgacc     3180

gtgaagcagc tgaaggagga ctacttcaag aagatcgagt gcttcgacag cgtggagatt     3240

tctggagtgg aggaccgctt caacgcatct ttggggacat accacgacct gctgaagatc     3300

atcaaggaca aggacttcct ggacaacgag gagaacgagg acatcctgga ggacattgtg     3360

ctgacactga ccctgttcga ggatagggag atgatcgagg agcgcctgaa gacatacgca     3420

cacctgtttg acgacaaggt gatgaagcag ctgaagagga ggcgctatac tggatgggga     3480

aggctgtcaa ggaagctgat taacgggatc cgcgacaagc agagcgggaa gacaattctg     3540

gacttcctga agagcgacgg gttcgcaaac cgcaacttca tgcagctgat ccacgacgat     3600

agcctgacct tcaaggagga catccagaag gcccaagtgt ctggacaagg ggatagcctg     3660

catgagcaca tcgcaaatct ggctgggtca cccgcaatca agaagggaat tctgcagacc     3720

gtgaaggtgg tggatgagct ggtgaaggtg atgggaaggc acaaacccga gaacatcgtg     3780

atcgagatgg caagggagaa ccagacaacc cagaagggac agaagaactc tagggagcgc     3840

atgaagcgca tcgaggaggg aattaaggag ctgggaagcc agatcctgaa ggagcatcct     3900

gtggagaaca cccaactgca gaacgagaag ctgtacctgt actacctgca gaacgggagg     3960

gacatgtacg tggatcaaga gctggacatc aaccgcctga gcgactatga cgtggaccac     4020

attgtgcctc agtcgttcct gaaggacgac agcatcgaca acaaggtgct gacaaggagc     4080

gacaagaatc gcggaaagag cgacaacgtg ccttcagaag aggtggtgaa gaagatgaag     4140

aactactggc gccagctgct gaacgcaaag ctgattacac agcgcaagtt cgacaacctg     4200

accaaggcag agaggggagg actgtcagaa ctggataagg ccgggttcat caagaggcaa     4260

ctggtggaga cacgccagat cacaaagcat gtggcccaga ttctggacag ccgcatgaac     4320

accaagtacg acgagaacga caagctgatc cgcgaggtga aggtgattac cctgaagagc     4380

aagctggtga gcgactttcg caaggacttc cagttctaca aggtgcgcga gatcaacaac     4440

taccaccacg cacacgacgc ctacctgaat gcagttgtgg gaacagccct gatcaagaag     4500

taccccaagc tggagagcga gttcgtgtat ggggactaca aggtgtacga cgtgcgcaag     4560

atgatcgcca agtctgagca agagatcggg aaggcaaccg ccaagtactt cttctacagc     4620

aacatcatga acttcttcaa gaccgagatc accctggcca atggggagat taggaagaga     4680

cccctgatcg agaccaacgg agagactgga gagatcgtgt gggataaggg gagggacttt     4740

gcaacagtgc gcaaagtgct gagcatgcct caagtgaaca tcgtgaagaa gaccgaggtg     4800

cagactgggg gattctcaaa ggagagcatt ctgcccaagc gcaacagcga taagctgatt     4860

gcacgcaaga aggactggga ccccaagaag tatggggggt ttgatagccc caccgtggca     4920

tattctgtgt tggttgtggc caaggtggag aaggggaaga gcaagaagct gaagagcgtg     4980

aaggagctgc tggggatcac cattatggag aggagcagct tcgagaagaa ccccatcgac     5040

ttcctggagg caaaggggta taaggaggtg aagaaggacc tgatcatcaa gctgcccaag     5100

tacagcctgt tcgagctgga gaatgggagg aagaggatgc tggcatctgc tggagaactg     5160

cagaagggga atgagttggc actgcctagc aagtacgtga acttcctgta cctggccagc     5220

cactacgaga agctgaaggg atcacccgag gacaatgagc agaagcagct gtttgtggag     5280

cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttcag caagcgcgtg     5340

attctggcag acgcaaacct ggataaggtg ctgagcgcct acaacaagca ccgcgataag     5400

cccattcgcg agcaagcaga gaacatcatc cacctgttca ccctgaccaa cctgggagca     5460

cctgcagcat tcaagtactt cgacaccacc atcgaccgca agaggtacac aagcaccaag     5520

gaagtgctgg acgcaaccct                                                 5540


<210>  45
<211>  23
<212>  DNA
<213>  Parachlorella sp.


<220>
<221>  misc_feature
<223>  SRP54 CRISPR target sequence (including PAM)

<400>  45
ggcgtgggac atggtgcgca agg                                               23


<210>  46
<211>  32
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic


<220>
<221>  misc_feature
<223>  AE596 Primer

<400>  46
tgcgacatgc agcttactaa cctgctcgac at                                     32


<210>  47
<211>  28
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic


<220>
<221>  misc_feature
<223>  AE597 Primer

<400>  47
atgggctcct tgatgttgtc cgccgtta                                          28


<210>  48
<211>  21
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic


<220>
<221>  misc_feature
<223>  AE405 Primer

<400>  48
acccaaaccc atgccagtgt a                                                 21


<210>  49
<211>  25
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic


<220>
<221>  misc_feature
<223>  AE406 Primer

<400>  49
actgtatgca gagtggtctg aagtg                                             25


<210>  50
<211>  570
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic


<220>
<221>  misc_feature
<223>  Nat1 gene encoding nourseothricin acetyltransferase from 
       Streptomyces noursei, codon-optimized for Parachlorella

<400>  50
atgaccacac tcgacgacac cgcatacaga taccgcacaa gcgttcctgg cgacgcagag       60

gcaattgagg cattagatgg cagcttcacc accgacaccg tgtttagagt gacagcaacc      120

ggcgacggct ttacacttag agaggttcca gtggacccgc cattgacaaa ggtgtttcca      180

gacgacgaga gcgacgatga gagcgatgct ggtgaagatg gcgacccaga tagccgcaca      240

tttgtggcat acggcgatga tggcgacctt gctgggtttg tggtggtgag ctactctggc      300

tggaatagac gcttgaccgt ggaggatatc gaggttgcac cagagcatag aggccatgga      360

gtgggtagag cactgatggg tctggcaacc gagtttgcaa gggagagagg tgctggtcat      420

ctgtggttgg aggtgacaaa cgtgaacgca ccagcgatcc acgcatacag aagaatgggc      480

ttcacgctgt gcggcctgga tacagcactg tatgatggca cagcgagcga tggtgagcaa      540

gcactgtaca tgagcatgcc atgcccgtaa                                       570


<210>  51
<211>  2862
<212>  DNA
<213>  Artificial Sequence

<220>
<223>  Synthetic


<220>
<221>  misc_feature
<223>  Nat1 gene Intronylated with Parachlorella RPS4 Introns

<400>  51
atgaccactt tgtgagttct gagaagctga ttgttgttta acttctttga aagctttatc       60

gaagattctg caagcgatga acattgcttg tcaagaccga gagctgcatg cccacttgac      120

atccagcttt gaacggctct tcatgtttga tttgtttctg attgtaggga cgacaccgca      180

tacagatacc gcacaagcgt tcctggcgac gcagaggcaa ttgaggcatt agatggcagc      240

ttcaccaccg acaccgtgtt tagagtgagt gcagcgtcag ctgtggcagt tgttggcttt      300

cgtctcagtc agtagtttgc tgggattgat tatggagggc acagttgcaa ttttgagttg      360

cacgttgcga caagcgtgtt gacaaagcgt ggtcaagccg gccagtcttg ccggtggcgg      420

gtggcttggt ctaacttccg ctctacagca atcgttttgt tcatggttac ggggctggcg      480

tgccagaaag tcctggtcag ccaccctcgc ttcaaagccg tagcccaaca actttgcgaa      540

tatgttcgat ttgcaggtaa cagcaaccgg cgacggcttt acacttagag aggttccagt      600

ggacccgcca ttgacaaagg tacagctctg cgtgcaacag gttgcaagat gcagcgcagg      660

tcttccctgg tcaaacgatg tatgcagagt tgagaggcac ttgagctggg tgaatggcgt      720

gggctcgtag gtagtgtgca gggcaggaag ggcagccaat tttggagttg tggtccggtg      780

tcgttgcttc gagccttatt aggactcttg ctcatcaaag cgttagttgt gaataagttg      840

atctgaaagg atgttatgta cagcaagcag cagcagttaa gagtctgggg agtagctgca      900

cagggcgagg tgtcaagatg ggaagggtcc tgcctcctta tgtgtttttc cctgtagggg      960

aggaagcctc ttatgggcaa tggttgggca tattttccag ccagcccttc tttctatagg     1020

ggccagggtg ggcccagctc gtcttggctt ccaccaccag gagagtgagg gcattgaagg     1080

gccataaata gtcctcccat ctacgtgcac cagagggtgt cgtctaggct gtgcatgcca     1140

cgaggggaag gagccaagaa tgagtgtatg ggttgttttc atgtttaggc tgggataaaa     1200

ctgttttcaa ttgcgcctgc cgggtgaaaa ccacagcagc atcagcaagc ttggagaagg     1260

ccagcccgcc cagcacaggc tcacgttccc actcaggcgg tcagtcgggc gggggtgtga     1320

gtcaggcagg cgagggtgtc tgtgcctgac atcagcacct ctgcttagcc actgcagccc     1380

ctggagcagg gtagggcgtc atttgcagca atcacctgct gcctcacacg tcgcagcttg     1440

gaatttcaac gaccatcagc gctggggttg ttgagggatc atagcagatt ttggtgcagc     1500

ctggttgtca tgctctttgt ggaatggcct ctatgttcga gcaattcgtt ggatgttgag     1560

gtgcttgggg acagagagtc gaatgatggg ccagggtcaa acatgcgagc gtttggctga     1620

gtcagcggtt tttgctggtc actttttctt ttgtttctta tttaggtttg atggatgtgt     1680

tttgtgctgc tgccctgaag ctgcagcagc gtgtctgccc tgcgctactg cgggcaccaa     1740

ggctatgtgc tggtgcactc ggctgcgctg cacctgtgca cctcgcactc cgtccagcct     1800

ccatgcagca cacgtactca cggtgtcctc ctgacctgtc gtacgctatt ccaaacttgc     1860

tcttttgctg ccgctgctct cgtacacaat tgctgttgat tatcgatatc taatcgagcg     1920

cctgctgact gaactccgca ggtgtttcca gacgacgaga gcgacgatga gagcgatgct     1980

ggtgaagatg gcgacccaga tagccgcaca tttgtggcat acggcgatga tggcgacctt     2040

gctgggtttg tggtggtgag ctactctggc tggaatagac gcttgaccgt ggaggatatc     2100

gaggttgcac cagagcatag aggccatgga gtgggtagag cactgatggg tctggcaacc     2160

gagtttgcaa ggtgggtggg ctctgaagga ggaggaggga gcgggtgatt aaacagggcc     2220

tgcatgaaga ggagcagggg ctgcatggac agcaggggga aggtgcagaa gggagggtca     2280

agcggggttc aggtggctgt gggtttctgc acgagcagtg aaagaagctg tatccttcca     2340

cctgctttca ctggcgaaag gttgaaaaca ggatgtcgca gctggaaaga tgttgcgctg     2400

tcaagtgcaa gccatggttg agggtatgcc tgtgtgcatg tgcttcttaa agttactcct     2460

gttctatggt tctgggtgct tgttgtttgt ggtgcaggga gagaggtgct ggtcatctgt     2520

ggttggaggt gacaaacgtg aacgcaccag cgatccacgc atacagaaga atgggcttca     2580

cgctgtgcgg cctggataca gcactgtatg atggtgaggg ggcatgtaag caatggcagg     2640

caattcaaga acgaatcatt gctgcaaatg ctgggatggt atgcagctga ggtatctatt     2700

gccttgtatt ttgtctcgca ttgcatcggt ggtgcgttct gtggcctgag gcacagttct     2760

tgctgtttga taagggttcg actgagttgt cgtgtgtgct gtgctgcagg cacagcgagc     2820

gatggtgagc aagcactgta catgagcatg ccatgcccgt aa                        2862


