LOCUS       sequence-202020-       10558 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cas9-Nlux-P2A-BlastR and sgRNA in Nannochloropsis 
            oceanica..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10558)
  AUTHORS   Poliner E, Takeuchi T, Du ZY, Benning C, Farre EM
  TITLE     Nontransgenic Marker-Free Gene Disruption by an Episomal CRISPR 
            System in the Oleaginous Microalga, Nannochloropsis oceanica 
            CCMP1779.
  JOURNAL   ACS Synth Biol. 2018 Apr 20;7(4):962-968. doi: 
            10.1021/acssynbio.7b00362. Epub 2018 Mar 14.
  PUBMED    29518315
REFERENCE   2  (bases 1 to 10558)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acssynbio.7b00362"; journalName: "ACS Synth Biol"; date: 
            "2018-04-20- 20"; volume: "7"; issue: "4"; pages: "962-968"
COMMENT     Sequence Label: pNOC-CRISPR-P2A-BlastR
FEATURES             Location/Qualifiers
     source          1..10558
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        22..126
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(127..195,196..987)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(345..364)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1158..1746
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1647..1666
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1900..1917
                     /label=L4440
                     /note="L4440 vector, forward primer"
     regulatory      2162..2171
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(3245..3301)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(3434..3454)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      3459..3468
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(3470..3496)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(4049..8149)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(4049..8149)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(4049..8149)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(8150..8170)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(8391..8402)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_RNA        9393..9468
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     ncRNA           9473..9540
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
ORIGIN
        1 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
       61 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
      121 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
      181 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
      241 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
      301 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
      361 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
      421 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
      481 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
      541 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
      601 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
      661 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
      721 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
      781 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
      841 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
      901 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
      961 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     1021 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     1081 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     1141 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     1201 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     1261 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     1321 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     1381 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     1441 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     1501 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     1561 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     1621 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     1681 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     1741 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     1801 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     1861 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     1921 gctggtcact ggattttggt tttaggaatt agaaatttta ttgatagaag tattttacaa
     1981 atacaaatac atactaaggg tttcttatat gctcaacaca tgagcgaaac cctataagaa
     2041 ccctaattcc cttatctggg aactactcac acattattct ggagaaaaat agagagagat
     2101 agatttgtag agagagactg gtgatttttg cggactccgg tcggcatcta ctactagtgc
     2161 ggccgccatg gcctctagtg gatcagcttg catgcctgca ggagacgata tcacctcttc
     2221 tgtttccacg ataaaaatag actgctcatt tcttcgtcgt cttcatcgtc tgctttttct
     2281 gcttcgcctc tgtctggggt ctgaaaccac tacacacaca cacaacactc gtactcccac
     2341 tttcacaaaa gcgtaagctc accggctttt cttacacgta cattttagtg gatcccatca
     2401 cgccactacc acgcccgcgg gggatggaac ggaggggaga gagagaaggg ggaagcatgg
     2461 atgaatgaga cattgaggga aaggagggga gggagcagtc catcagggcg ctacctctct
     2521 tgtcccccaa accctgttga gccgttcaac atgtttcatg tttcctccct ccccccttcc
     2581 ctccctggcc tttccgcgga gccattcaag tgacgtctgg accgcaccgt aacaaaatcg
     2641 tttctatggg gggtttgttt gacaaccacg tcttcagcgt ttttaaaaaa aaaagcgggc
     2701 aagccctctc accctcactc atgcccatcc tcctcctctc ctgcggaaca ttcttacaaa
     2761 aggcgtaact cgacgacaac tcaaagaacg acaaacatca atcccaaaaa aaaaatctct
     2821 actcgtctct cttggatctt tccaattgac tagttcactc ccacacgtag cccgaaggga
     2881 gaagttctct gataccgacc gccgtaggct gcccgtcgga atccttcaca atcgccttga
     2941 tcccagggtg gaggtcgagc aggacctgcc ggcaccgccc gcaggggctc agaatcccgc
     3001 ggttctcgtt cccgatggcg acaatgcacg tcaggttccc agccgcggca gcagcggcgg
     3061 tccccaggac caccagctcc gcgcaaggcc cccccgtaaa atggtacacg ttgacccccg
     3121 tgaagatccg tccgtcggac gacagagcag cggaagccac cgagtaatcc tcggagatag
     3181 gaatcgagtt aatcgtcgcc gtagccctct cgatcagcgt gctctcctcc tgcgagaggg
     3241 gcatgggccc cgggttctcc tccacgtccc cggcctgctt cagcaaggag aagttcgtcg
     3301 cccccggccc ctggaaggac agggtcgtca tcgcggtgct caggcgttcg ctgcggccat
     3361 cgctgttttc cggatggctt ttgctaaacg ccgccggatc atgaatgccg gggcgcgggc
     3421 aaaagccgct tccgaccttc ctcttctttt tcgggctagc caccatggac gcgtagtcgg
     3481 gcacgtcgta ggggtatccg gccaggatgc gctcgcacag gcgccagccg gttaccccgt
     3541 tgatggtcac gcggaacaac agcgagccat ccgggttgat cagccgctcg tcaatgatct
     3601 tattgccgtt ccacagggtg ccggtcaccg tgatcttctt gccgtcgaac acggcaatgc
     3661 cctcgtaagg acggccgaag tagtcaatca tgttcggcgt cacaccgtcg atgaccagcg
     3721 tcccgtagtg cagtatcacc ttgaagtggt gatcgtccac ggggtacaca accttgaaga
     3781 tcttctcgat ctggcccatt tggtccccgc tcaggccctc gtagggaatg atgacgtgga
     3841 tgtcgatctt caggccgttc tcgccgctca ggacgatgcg ctggattggg gtcacggaga
     3901 cgcccaggtt ctggaaaagc gaggacacac cgccctgctc cagcacctgg tcgaggttat
     3961 agccagctgt ctgccgccag tcgcccacga agtcctcgag agtaaacacc atgttaacgg
     4021 acccgctgcc ggacccgtca gccctgctgt ctccaccgag ctgagagagg tcgattcttg
     4081 tttcatagag ccccgtaatt gactgatgaa tcagtgtggc gtccaggacc tcctttgtag
     4141 aggtgtaccg ctttctgtct atggtggtgt cgaagtactt gaaggctgca ggcgcgccca
     4201 agttggtcag agtaaacaag tggataatgt tttctgcctg ctccctgatg ggcttatccc
     4261 tgtgcttatt gtaagcagaa agcaccttat cgaggttagc gtcggcgagg atcactcttt
     4321 tggagaattc gcttatttgc tcgatgatct catcaaggta gtgtttgtgt tgttccacga
     4381 acagctgctt ctgctcatta tcttcgggag accctttgag cttttcatag tggctggcca
     4441 gatacaagaa attaacgtat ttagagggca gtgccagctc gttacctttc tgcagctcgc
     4501 ccgcactagc gagcattcgt ttccggccgt tttcaagctc aaagagagag tacttgggaa
     4561 gcttaatgat gaggtctttt ttgacctctt tatatccttt cgcctcgaga aagtcgatgg
     4621 ggtttttttc gaagcttgat cgctccatga ttgtgatgcc cagcagttcc ttgacgcttt
     4681 tgagtttttt agacttccct ttctccactt tggccacaac cagtacactg taagcgactg
     4741 taggagaatc gaatccgccg tatttcttgg ggtcccaatc ttttttgcgt gcgatcagct
     4801 tgtcgctgtt ccttttcggg aggatacttt ccttggagaa gcctccggtc tgtacttcgg
     4861 tctttttaac gatgttcacc tgcggcatgg acaggacctt ccggactgtc gcgaaatccc
     4921 tacccttgtc ccacacgatt tctcctgttt ctccgtttgt ttcgataagt ggtcgcttcc
     4981 gaatctctcc attggccagt gtaatctcgg tcttgaaaaa attcataata ttgctgtaaa
     5041 agaagtactt agcggtggcc ttgcctattt cctgctcaga ctttgcgatc attttcctaa
     5101 catcgtacac tttatagtct ccgtaaacaa attcagattc aagcttggga tattttttga
     5161 taagtgcagt gcctaccact gcattcaggt aggcatcatg cgcatggtgg taattgttga
     5221 tctctctcac cttataaaac tgaaagtcct ttctgaaatc tgagaccagc ttagacttca
     5281 gagtaataac tttcacctct cgaatcagtt tgtcattttc atcgtacttg gtgttcatgc
     5341 gtgaatcgag aatttgggcc acgtgcttgg tgatctggcg tgtctcaaca agctgccttt
     5401 tgatgaagcc ggctttatcc aactcagaca ggccacctcg ttcagcctta gtcagattat
     5461 cgaacttccg ttgtgtgatc agtttggcgt tcagcagctg ccgccaataa tttttcattt
     5521 tcttgacaac ttcttctgag gggacgttat cactcttccc tctattttta tcggatcttg
     5581 tcaacacttt attatcaata gaatcatctt tgagaaaaga ctggggcacg atatgatcca
     5641 cgtcgtagtc ggagagccga ttgatgtcca gttcctgatc cacgtacatg tccctgccgt
     5701 tctgcaggta gtacaggtag agcttctcat tctgaagctg ggtgttttca actgggtgtt
     5761 ccttaaggat ttgggacccc agttctttta taccctcttc aatcctcttc atcctttccc
     5821 tactgttctt ctgtcccttc tgggtagttt ggttctctcg ggccatctcg ataacgatat
     5881 tctcgggctt atgccttccc attactttga cgagttcatc cacgacctta acggtctgca
     5941 gtattccctt tttgatagct gggctacctg caagattagc gatgtgctcg tgaagactgt
     6001 ccccctggcc agaaacttgt gctttctgga tgtcctcctt aaaggtgaga gagtcatcat
     6061 ggatcaactg catgaagttc cggttggcaa atccatcgga cttaagaaaa tccaggattg
     6121 tctttccact ctgcttgtct cggatcccat tgatcagttt tcttgacagc cgcccccatc
     6181 ctgtatatcg gcgcctcttg agctgtttca tgactttgtc gtcgaagaga tgagcgtaag
     6241 ttttcaagcg ttcttcaatc atctccctat cttcaaacaa cgtaagggtg aggacaatgt
     6301 cctcaagaat gtcctcgttc tcctcattgt ccaggaagtc cttgtcttta atgattttca
     6361 ggagatcgtg atacgttccc agggatgcgt tgaagcgatc ctccactccg ctgatttcaa
     6421 cagagtcgaa acattcaatc tttttgaaat agtcttcttt gagctgtttc acggtaactt
     6481 tccggttcgt cttgaagagg aggtccacga tagctttctt ctgctctcca gacaggaatg
     6541 ctggctttct catcccttct gtgacgtatt tgaccttggt gagctcgtta taaactgtga
     6601 agtactcgta cagcagagag tgtttaggaa gcaccttttc gttaggcaga tttttatcaa
     6661 agttagtcat cctttcgatg aaggactggg cagaggcccc cttatccacg acttcctcga
     6721 agttccaggg agtgatggtc tcttctgatt tgcgagtcat ccacgcgaat ctggaatttc
     6781 cccgggcgag ggggcctaca tagtagggta tccgaaatgt gaggattttc tcaatctttt
     6841 ccctgttatc tttcaaaaag gggtagaaat cctcttgccg cctgaggata gcgtgcagtt
     6901 cgcccaggtg aatctggtgg gggatgcttc cattgtcgaa agtgcgctgt ttgcgcaaca
     6961 gatcttctct gttaagcttt accagcagct cctcggtgcc gtccattttt tccaagatgg
     7021 gcttaataaa tttgtaaaat tcctcctggc ttgctccgcc gtcaatgtat ccggcgtagc
     7081 catttttaga ctgatcgaag aaaatttcct tgtacttctc aggcagttgc tgtctgacaa
     7141 gggccttcag caaagtcaag tcttggtggt gctcatcata gcgcttgatc atactagcgc
     7201 tcagcggagc tttggtgatc tccgtgttca ctcgcagaat atcactcagc agaatggcgt
     7261 ctgacaggtt ctttgccgcc aaaaaaaggt ctgcgtactg gtcgccgatc tgggccagca
     7321 gattgtcgag atcatcatcg taggtgtctt tgctcagttg aagcttggca tcttcggcca
     7381 ggtcgaagtt agatttaaag ttgggggtca gcccgagtga cagggcgata agattaccaa
     7441 acaggccgtt cttcttctcc ccagggagct gtgcgatgag gttttcgagc cgccgggatt
     7501 tggacagcct agcgctcagg attgctttgg cgtcaactcc ggatgcgttg atcgggttct
     7561 cttcgaaaag ctgattgtaa gtctgaacca gttggataaa gagtttgtcg acatcgctgt
     7621 tgtctgggtt caggtccccc tcgatgagga agtgtccccg aaatttgatc atatgcgcca
     7681 gcgcgagata gatcaaccgc aagtcagcct tatcagtact gtctacaagc ttcttcctca
     7741 gatgatatat ggttgggtac ttttcatggt acgccacctc gtccacgata ttgccaaaga
     7801 ttgggtggcg ctcgtgcttt ttatcctcct ccaccaaaaa ggactcctcc agcctatgga
     7861 agaaagagtc atccacctta gccatctcat tactaaagat ctcctgcagg tagcagatcc
     7921 gattctttct gcgggtatat ctgcgccgtg ctgttctttt gagccgcgtg gcttcggccg
     7981 tctccccgga gtcgaacagg agggcgccaa tgaggttctt ctttatgctg tggcgatcgg
     8041 tattgcccag aactttgaat tttttgctcg gcaccttgta ctcgtccgta atgacggccc
     8101 agccgacgct gtttgtgccg atatcgagcc caatggagta cttcttgtcc accttccgct
     8161 ttttcttggg catgctcgag ggttgcgtgt gtatctgtgt gcagtggata ttgttaccga
     8221 gtttggtgag cgtgagtccg ttagtgccct ggtggtggtg gattaggaga gtgggtgact
     8281 cggtgtccat ggctttcttc gctcattata ggaggggaaa ggaatgaggg agggtgggga
     8341 gaccgcgtct gttgttgacc accgatttac ttcttgcctc ccttcccctc cctcccctca
     8401 atccgtacga cacaaatagt agccgagtgt ctgctgcaga gcgcatgatt agtgtggtag
     8461 acaacgaggg agggaaggat gtacagggca tggcacggag aagcgatggt ggccaggaag
     8521 aggagaggtc gcgagaacag gatgtgttgc gaatggataa aaacagaaag cgatggctct
     8581 gggcttcgaa agcaggggac attaggacgt gtagaccatc tcgacggatc cctctgtatc
     8641 tctgttgtgc gtgaatgttt tctgtgcacg tgtagtgtgt gagagtagaa cccgggaact
     8701 cgaacagaga aaagcatggg tggctgtggt gtggaggctt cgttcccacc acatgccctt
     8761 ctccttcgcc tcgcctctcc ctgccttctt ccacgcaccc ttgcgcccct cgttctcaat
     8821 acctggctca cttccaccat tcaaacaacc atcacgatac aggcatttat ctatcgttga
     8881 agacttcttc ctccggtaga tcttagccaa ggtaagaaga ggggcatgca gcaaggagaa
     8941 agaaatgatg catgatgagg aatagaaggg gaggagggag ggatatgatg ggaagcgaaa
     9001 gcgcatattc tggtggtctg ctgcctgatg gggacgcgtc tagctgtgac actgaggacg
     9061 gtggctgctg gtggctgcgg gcgctgcctt ggtgatcaat gggagtaaag ggagggaagg
     9121 aggtagcgtg aacggatgac gcggagaagt ttaggggtct ctttacgtat cgcccctgcc
     9181 gcccgcctct ctgcgataaa tgtgcctgtt accctgcagc ctctattctt cactgtgttc
     9241 ctgttttcca acagcctcta ttcttccctg tcttttgttg cagtggcgtc atcctctctt
     9301 tgccccagtc gtcgttctct cgactcactc actccccccc tccttccctc cctccatcca
     9361 cagaatcgat gaagagcagg ccttggctct tcgttttaga gctagaaata gcaagttaaa
     9421 ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt ttggccggca
     9481 tggtcccagc ctcctcgctg gcgccggctg ggcaacatgc ttcggcatgg cgaatgggac
     9541 tcctgggtac catgggaaag aaaggatgag aaaggagaaa ggacatctag ataaccggca
     9601 tatcacggtg gtgtattagt gtagaatagt gaagagaaga cttgggaaaa tgtgtaggaa
     9661 aggttgtttc tgtgtatgtg ggttgggatg ggtggctgtt tgagaaggaa cagcgggcag
     9721 ggcgatgtag tgctgaacgg gcacggaacc actgagactg aaggaagtag ggagagagag
     9781 gggcagggga cgtgcacttt aatctttgcc tcggtagagt atacccatgc aagagtatgt
     9841 ggccacctgt ggtggctttt ggccaggtct ggtgcagtgc caatcatctc ccatcaataa
     9901 tacaacttca gaacaacggc gcattgatgg ggagagagaa agtaaattta agtaaggggt
     9961 acgtagtaga ggattcaact gaaatatttt cgaggagcgg ttgggaagtt gaccgattga
    10021 aaggagaagg gaggggagca ggtgtgatag tcatgtgtaa agtaattctt ttttgccgtc
    10081 gtcacacaat ccacatcaat gataaaatat gtttaaggat caatcacacg gagtcggtca
    10141 taaggcaacc gcaaacgcaa tgcaaactag caagcaagca ggaccacaac aacaaccatt
    10201 accatcacag gcgacagcag cagcagctgc agcagcagca aggcaagcaa aggccatctg
    10261 cgcgtggact tttccaggca ccggctttaa gcgtaatttt caatgattgt tgtccgtgta
    10321 tcctctcacc ctttcaccgc ttgcgcgcac gtgagcagca caactggtaa tgcccgagga
    10381 caatagacga ggaaaaagaa gaagaaaaat gaagagggag tgatgaagaa gaagagaaga
    10441 ataaaagaag acagaaaaga agataaaaaa gtcaaaagac gacaaagacg gaaaaaaaag
    10501 aaacgcgcaa ttttaacacc agcaacgacg acgaaaaggg cgtcagcttt gcaccggt
//
