LOCUS       sequence-182798-       10560 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 10560)
  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 10560)
  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..10560
                     /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      2164..2173
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(3247..3303)
                     /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(3436..3456)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      3461..3470
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(3472..3498)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(4051..8151)
                     /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(4051..8151)
                     /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(4051..8151)
                     /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(8152..8172)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(8393..8404)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_RNA        9395..9470
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     ncRNA           9475..9542
                     /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 gctcaggtca ctggattttg gttttaggaa ttagaaattt tattgataga agtattttac
     1981 aaatacaaat acatactaag ggtttcttat atgctcaaca catgagcgaa accctataag
     2041 aaccctaatt cccttatctg ggaactactc acacattatt ctggagaaaa atagagagag
     2101 atagatttgt agagagagac tggtgatttt tgcggactcc ggtcggcatc tactactagt
     2161 gcggccgcca tggcctctag tggatcagct tgcatgcctg caggagacga tatcacctct
     2221 tctgtttcca cgataaaaat agactgctca tttcttcgtc gtcttcatcg tctgcttttt
     2281 ctgcttcgcc tctgtctggg gtctgaaacc actacacaca cacacaacac tcgtactccc
     2341 actttcacaa aagcgtaagc tcaccggctt ttcttacacg tacattttag tggatcccat
     2401 cacgccacta ccacgcccgc gggggatgga acggagggga gagagagaag ggggaagcat
     2461 ggatgaatga gacattgagg gaaaggaggg gagggagcag tccatcaggg cgctacctct
     2521 cttgtccccc aaaccctgtt gagccgttca acatgtttca tgtttcctcc ctcccccctt
     2581 ccctccctgg cctttccgcg gagccattca agtgacgtct ggaccgcacc gtaacaaaat
     2641 cgtttctatg gggggtttgt ttgacaacca cgtcttcagc gtttttaaaa aaaaaagcgg
     2701 gcaagccctc tcaccctcac tcatgcccat cctcctcctc tcctgcggaa cattcttaca
     2761 aaaggcgtaa ctcgacgaca actcaaagaa cgacaaacat caatcccaaa aaaaaaatct
     2821 ctactcgtct ctcttggatc tttccaattg actagttcac tcccacacgt agcccgaagg
     2881 gagaagttct ctgataccga ccgccgtagg ctgcccgtcg gaatccttca caatcgcctt
     2941 gatcccaggg tggaggtcga gcaggacctg ccggcaccgc ccgcaggggc tcagaatccc
     3001 gcggttctcg ttcccgatgg cgacaatgca cgtcaggttc ccagccgcgg cagcagcggc
     3061 ggtccccagg accaccagct ccgcgcaagg cccccccgta aaatggtaca cgttgacccc
     3121 cgtgaagatc cgtccgtcgg acgacagagc agcggaagcc accgagtaat cctcggagat
     3181 aggaatcgag ttaatcgtcg ccgtagccct ctcgatcagc gtgctctcct cctgcgagag
     3241 gggcatgggc cccgggttct cctccacgtc cccggcctgc ttcagcaagg agaagttcgt
     3301 cgcccccggc ccctggaagg acagggtcgt catcgcggtg ctcaggcgtt cgctgcggcc
     3361 atcgctgttt tccggatggc ttttgctaaa cgccgccgga tcatgaatgc cggggcgcgg
     3421 gcaaaagccg cttccgacct tcctcttctt tttcgggcta gccaccatgg acgcgtagtc
     3481 gggcacgtcg taggggtatc cggccaggat gcgctcgcac aggcgccagc cggttacccc
     3541 gttgatggtc acgcggaaca acagcgagcc atccgggttg atcagccgct cgtcaatgat
     3601 cttattgccg ttccacaggg tgccggtcac cgtgatcttc ttgccgtcga acacggcaat
     3661 gccctcgtaa ggacggccga agtagtcaat catgttcggc gtcacaccgt cgatgaccag
     3721 cgtcccgtag tgcagtatca ccttgaagtg gtgatcgtcc acggggtaca caaccttgaa
     3781 gatcttctcg atctggccca tttggtcccc gctcaggccc tcgtagggaa tgatgacgtg
     3841 gatgtcgatc ttcaggccgt tctcgccgct caggacgatg cgctggattg gggtcacgga
     3901 gacgcccagg ttctggaaaa gcgaggacac accgccctgc tccagcacct ggtcgaggtt
     3961 atagccagct gtctgccgcc agtcgcccac gaagtcctcg agagtaaaca ccatgttaac
     4021 ggacccgctg ccggacccgt cagccctgct gtctccaccg agctgagaga ggtcgattct
     4081 tgtttcatag agccccgtaa ttgactgatg aatcagtgtg gcgtccagga cctcctttgt
     4141 agaggtgtac cgctttctgt ctatggtggt gtcgaagtac ttgaaggctg caggcgcgcc
     4201 caagttggtc agagtaaaca agtggataat gttttctgcc tgctccctga tgggcttatc
     4261 cctgtgctta ttgtaagcag aaagcacctt atcgaggtta gcgtcggcga ggatcactct
     4321 tttggagaat tcgcttattt gctcgatgat ctcatcaagg tagtgtttgt gttgttccac
     4381 gaacagctgc ttctgctcat tatcttcggg agaccctttg agcttttcat agtggctggc
     4441 cagatacaag aaattaacgt atttagaggg cagtgccagc tcgttacctt tctgcagctc
     4501 gcccgcacta gcgagcattc gtttccggcc gttttcaagc tcaaagagag agtacttggg
     4561 aagcttaatg atgaggtctt ttttgacctc tttatatcct ttcgcctcga gaaagtcgat
     4621 ggggtttttt tcgaagcttg atcgctccat gattgtgatg cccagcagtt ccttgacgct
     4681 tttgagtttt ttagacttcc ctttctccac tttggccaca accagtacac tgtaagcgac
     4741 tgtaggagaa tcgaatccgc cgtatttctt ggggtcccaa tcttttttgc gtgcgatcag
     4801 cttgtcgctg ttccttttcg ggaggatact ttccttggag aagcctccgg tctgtacttc
     4861 ggtcttttta acgatgttca cctgcggcat ggacaggacc ttccggactg tcgcgaaatc
     4921 cctacccttg tcccacacga tttctcctgt ttctccgttt gtttcgataa gtggtcgctt
     4981 ccgaatctct ccattggcca gtgtaatctc ggtcttgaaa aaattcataa tattgctgta
     5041 aaagaagtac ttagcggtgg ccttgcctat ttcctgctca gactttgcga tcattttcct
     5101 aacatcgtac actttatagt ctccgtaaac aaattcagat tcaagcttgg gatatttttt
     5161 gataagtgca gtgcctacca ctgcattcag gtaggcatca tgcgcatggt ggtaattgtt
     5221 gatctctctc accttataaa actgaaagtc ctttctgaaa tctgagacca gcttagactt
     5281 cagagtaata actttcacct ctcgaatcag tttgtcattt tcatcgtact tggtgttcat
     5341 gcgtgaatcg agaatttggg ccacgtgctt ggtgatctgg cgtgtctcaa caagctgcct
     5401 tttgatgaag ccggctttat ccaactcaga caggccacct cgttcagcct tagtcagatt
     5461 atcgaacttc cgttgtgtga tcagtttggc gttcagcagc tgccgccaat aatttttcat
     5521 tttcttgaca acttcttctg aggggacgtt atcactcttc cctctatttt tatcggatct
     5581 tgtcaacact ttattatcaa tagaatcatc tttgagaaaa gactggggca cgatatgatc
     5641 cacgtcgtag tcggagagcc gattgatgtc cagttcctga tccacgtaca tgtccctgcc
     5701 gttctgcagg tagtacaggt agagcttctc attctgaagc tgggtgtttt caactgggtg
     5761 ttccttaagg atttgggacc ccagttcttt tataccctct tcaatcctct tcatcctttc
     5821 cctactgttc ttctgtccct tctgggtagt ttggttctct cgggccatct cgataacgat
     5881 attctcgggc ttatgccttc ccattacttt gacgagttca tccacgacct taacggtctg
     5941 cagtattccc tttttgatag ctgggctacc tgcaagatta gcgatgtgct cgtgaagact
     6001 gtccccctgg ccagaaactt gtgctttctg gatgtcctcc ttaaaggtga gagagtcatc
     6061 atggatcaac tgcatgaagt tccggttggc aaatccatcg gacttaagaa aatccaggat
     6121 tgtctttcca ctctgcttgt ctcggatccc attgatcagt tttcttgaca gccgccccca
     6181 tcctgtatat cggcgcctct tgagctgttt catgactttg tcgtcgaaga gatgagcgta
     6241 agttttcaag cgttcttcaa tcatctccct atcttcaaac aacgtaaggg tgaggacaat
     6301 gtcctcaaga atgtcctcgt tctcctcatt gtccaggaag tccttgtctt taatgatttt
     6361 caggagatcg tgatacgttc ccagggatgc gttgaagcga tcctccactc cgctgatttc
     6421 aacagagtcg aaacattcaa tctttttgaa atagtcttct ttgagctgtt tcacggtaac
     6481 tttccggttc gtcttgaaga ggaggtccac gatagctttc ttctgctctc cagacaggaa
     6541 tgctggcttt ctcatccctt ctgtgacgta tttgaccttg gtgagctcgt tataaactgt
     6601 gaagtactcg tacagcagag agtgtttagg aagcaccttt tcgttaggca gatttttatc
     6661 aaagttagtc atcctttcga tgaaggactg ggcagaggcc cccttatcca cgacttcctc
     6721 gaagttccag ggagtgatgg tctcttctga tttgcgagtc atccacgcga atctggaatt
     6781 tccccgggcg agggggccta catagtaggg tatccgaaat gtgaggattt tctcaatctt
     6841 ttccctgtta tctttcaaaa aggggtagaa atcctcttgc cgcctgagga tagcgtgcag
     6901 ttcgcccagg tgaatctggt gggggatgct tccattgtcg aaagtgcgct gtttgcgcaa
     6961 cagatcttct ctgttaagct ttaccagcag ctcctcggtg ccgtccattt tttccaagat
     7021 gggcttaata aatttgtaaa attcctcctg gcttgctccg ccgtcaatgt atccggcgta
     7081 gccattttta gactgatcga agaaaatttc cttgtacttc tcaggcagtt gctgtctgac
     7141 aagggccttc agcaaagtca agtcttggtg gtgctcatca tagcgcttga tcatactagc
     7201 gctcagcgga gctttggtga tctccgtgtt cactcgcaga atatcactca gcagaatggc
     7261 gtctgacagg ttctttgccg ccaaaaaaag gtctgcgtac tggtcgccga tctgggccag
     7321 cagattgtcg agatcatcat cgtaggtgtc tttgctcagt tgaagcttgg catcttcggc
     7381 caggtcgaag ttagatttaa agttgggggt cagcccgagt gacagggcga taagattacc
     7441 aaacaggccg ttcttcttct ccccagggag ctgtgcgatg aggttttcga gccgccggga
     7501 tttggacagc ctagcgctca ggattgcttt ggcgtcaact ccggatgcgt tgatcgggtt
     7561 ctcttcgaaa agctgattgt aagtctgaac cagttggata aagagtttgt cgacatcgct
     7621 gttgtctggg ttcaggtccc cctcgatgag gaagtgtccc cgaaatttga tcatatgcgc
     7681 cagcgcgaga tagatcaacc gcaagtcagc cttatcagta ctgtctacaa gcttcttcct
     7741 cagatgatat atggttgggt acttttcatg gtacgccacc tcgtccacga tattgccaaa
     7801 gattgggtgg cgctcgtgct ttttatcctc ctccaccaaa aaggactcct ccagcctatg
     7861 gaagaaagag tcatccacct tagccatctc attactaaag atctcctgca ggtagcagat
     7921 ccgattcttt ctgcgggtat atctgcgccg tgctgttctt ttgagccgcg tggcttcggc
     7981 cgtctccccg gagtcgaaca ggagggcgcc aatgaggttc ttctttatgc tgtggcgatc
     8041 ggtattgccc agaactttga attttttgct cggcaccttg tactcgtccg taatgacggc
     8101 ccagccgacg ctgtttgtgc cgatatcgag cccaatggag tacttcttgt ccaccttccg
     8161 ctttttcttg ggcatgctcg agggttgcgt gtgtatctgt gtgcagtgga tattgttacc
     8221 gagtttggtg agcgtgagtc cgttagtgcc ctggtggtgg tggattagga gagtgggtga
     8281 ctcggtgtcc atggctttct tcgctcatta taggagggga aaggaatgag ggagggtggg
     8341 gagaccgcgt ctgttgttga ccaccgattt acttcttgcc tcccttcccc tccctcccct
     8401 caatccgtac gacacaaata gtagccgagt gtctgctgca gagcgcatga ttagtgtggt
     8461 agacaacgag ggagggaagg atgtacaggg catggcacgg agaagcgatg gtggccagga
     8521 agaggagagg tcgcgagaac aggatgtgtt gcgaatggat aaaaacagaa agcgatggct
     8581 ctgggcttcg aaagcagggg acattaggac gtgtagacca tctcgacgga tccctctgta
     8641 tctctgttgt gcgtgaatgt tttctgtgca cgtgtagtgt gtgagagtag aacccgggaa
     8701 ctcgaacaga gaaaagcatg ggtggctgtg gtgtggaggc ttcgttccca ccacatgccc
     8761 ttctccttcg cctcgcctct ccctgccttc ttccacgcac ccttgcgccc ctcgttctca
     8821 atacctggct cacttccacc attcaaacaa ccatcacgat acaggcattt atctatcgtt
     8881 gaagacttct tcctccggta gatcttagcc aaggtaagaa gaggggcatg cagcaaggag
     8941 aaagaaatga tgcatgatga ggaatagaag gggaggaggg agggatatga tgggaagcga
     9001 aagcgcatat tctggtggtc tgctgcctga tggggacgcg tctagctgtg acactgagga
     9061 cggtggctgc tggtggctgc gggcgctgcc ttggtgatca atgggagtaa agggagggaa
     9121 ggaggtagcg tgaacggatg acgcggagaa gtttaggggt ctctttacgt atcgcccctg
     9181 ccgcccgcct ctctgcgata aatgtgcctg ttaccctgca gcctctattc ttcactgtgt
     9241 tcctgttttc caacagcctc tattcttccc tgtcttttgt tgcagtggcg tcatcctctc
     9301 tttgccccag tcgtcgttct ctcgactcac tcactccccc cctccttccc tccctccatc
     9361 cacagaatcg atgaagagca ggccttggct cttcgtttta gagctagaaa tagcaagtta
     9421 aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc ttttggccgg
     9481 catggtccca gcctcctcgc tggcgccggc tgggcaacat gcttcggcat ggcgaatggg
     9541 actcctgggt accatgggaa agaaaggatg agaaaggaga aaggacatct agataaccgg
     9601 catatcacgg tggtgtatta gtgtagaata gtgaagagaa gacttgggaa aatgtgtagg
     9661 aaaggttgtt tctgtgtatg tgggttggga tgggtggctg tttgagaagg aacagcgggc
     9721 agggcgatgt agtgctgaac gggcacggaa ccactgagac tgaaggaagt agggagagag
     9781 aggggcaggg gacgtgcact ttaatctttg cctcggtaga gtatacccat gcaagagtat
     9841 gtggccacct gtggtggctt ttggccaggt ctggtgcagt gccaatcatc tcccatcaat
     9901 aatacaactt cagaacaacg gcgcattgat ggggagagag aaagtaaatt taagtaaggg
     9961 gtacgtagta gaggattcaa ctgaaatatt ttcgaggagc ggttgggaag ttgaccgatt
    10021 gaaaggagaa gggaggggag caggtgtgat agtcatgtgt aaagtaattc ttttttgccg
    10081 tcgtcacaca atccacatca atgataaaat atgtttaagg atcaatcaca cggagtcggt
    10141 cataaggcaa ccgcaaacgc aatgcaaact agcaagcaag caggaccaca acaacaacca
    10201 ttaccatcac aggcgacagc agcagcagct gcagcagcag caaggcaagc aaaggccatc
    10261 tgcgcgtgga cttttccagg caccggcttt aagcgtaatt ttcaatgatt gttgtccgtg
    10321 tatcctctca ccctttcacc gcttgcgcgc acgtgagcag cacaactggt aatgcccgag
    10381 gacaatagac gaggaaaaag aagaagaaaa atgaagaggg agtgatgaag aagaagagaa
    10441 gaataaaaga agacagaaaa gaagataaaa aagtcaaaag acgacaaaga cggaaaaaaa
    10501 agaaacgcgc aattttaaca ccagcaacga cgacgaaaag ggcgtcagct ttgcaccggt
//
