LOCUS       sequence-202029-       12242 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cas9-Nlux and sgRNA. Compact backbone contains CEN/ARS. 
            Hygromycin selection marker..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12242)
  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 12242)
  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-ARS-CRISPR-v2
FEATURES             Location/Qualifiers
     source          1..12242
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    17..520
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        557..661
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(662..730,731..1522)
                     /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(880..899)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1693..2281
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2182..2201
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2617..3642)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(2617..3642)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     regulatory      4434..4443
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(5124..5144)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      5149..5158
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(5160..5186)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(5739..9839)
                     /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(5739..9839)
                     /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(5739..9839)
                     /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(9840..9860)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(10081..10092)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_RNA        11083..11158
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     ncRNA           11163..11230
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
ORIGIN
        1 accggtttct tagacggatc gcttgcctgt aacttacacg cgcctcgtat cttttaatga
       61 tggaataatt tgggaattta ctctgtgttt atttattttt atgttttgta tttggatttt
      121 agaaagtaaa taaagaaggt agaagagtta cggaatgaag aaaaaaaaat aaacaaaggt
      181 ttaaaaaatt tcaacaaaaa gcgtacttta catatatatt tattagacaa gaaaagcaga
      241 ttaaatagat atacattcga ttaacgataa gtaaaatgta aaatcacagg attttcgtgt
      301 gtggtcttct acacagacaa gatgaaacaa ttcggcatta atacctgaga gcaggaagta
      361 caagataaaa ggtagtattt gttggcgatc cccctagagt cttttacatc ttcggaaaac
      421 aaaaactatt ttttctttaa tttctttttt tactttctat ttttaattta tatatttata
      481 ttaaaaaatt taaattataa ttatttttat agcacgtgat gaaaaggacc caggtggcac
      541 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
      601 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
      661 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
      721 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
      781 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
      841 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
      901 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
      961 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     1021 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     1081 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     1141 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     1201 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     1261 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     1321 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     1381 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     1441 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     1501 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     1561 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     1621 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     1681 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     1741 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     1801 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     1861 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     1921 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     1981 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     2041 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     2101 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     2161 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     2221 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     2281 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     2341 gttctttcct gcgttatccc ctgattctgt ggtttaaacc aggtcactgg attttggttt
     2401 taggaattag aaattttatt gatagaagta ttttacaaat acaaatacat actaagggtt
     2461 tcttatatgc tcaacacatg agcgaaaccc tataagaacc ctaattccct tatctgggaa
     2521 ctactcacac attattctgg agaaaaatag agagagatag atttgtagag agagactggt
     2581 gatttttgcg gactccggtc ggcatctact actagcctat tcctttgccc tcggacgagt
     2641 gctggggcgt cggtttccac tatcggcgag tacttctaca cagccatcgg tccagacggc
     2701 cgcgcttctg cgggcgattt gtgtacgccc gacagtcccg gctccggatc ggacgattgc
     2761 gtcgcatcga ccctgcgccc aagctgcatc atcgaaattg ccgtcaacca agctctgata
     2821 gagttggtca agaccaatgc ggagcatata cgcccggagc cgcggcgatc ctgcaagctc
     2881 cggatgcctc cgctcgaagt agcgcgtctg ctgctccata caagccaacc acggcctcca
     2941 gaagaagatg ttggcgacct cgtattggga atccccgaac atcgcctcgc tccagtcaat
     3001 gaccgctgtt atgcggccat tgtccgtcag gacattgttg gagccgaaat ccgcgtgcac
     3061 gaggtgccgg acttcggggc agtcctcggc ccaaagcatc agctcatcga gagcctgcgc
     3121 gacggacgca ctgacggtgt cgtccatcac agtttgccag tgatacacat ggggatcagc
     3181 aatcgcgcat atgaaatcac gccatgtagt gtattgaccg attccttgcg gtccgaatgg
     3241 gccgaacccg ctcgtctggc taagatcggc cgcagcgatc gcatccatgg cctccgcgac
     3301 cggctgcaga acagcgggca gttcggtttc aggcaggtct tgcaacgtga caccctgtgc
     3361 acggcgggag atgcaatagg tcaggctctc gctgaattcc ccaatgtcaa gcacttccgg
     3421 aatcgggagc gcggccgatg caaagtgccg ataaacataa cgatctttgt agaaaccatc
     3481 ggcgcagcta tttacccgca ggacatatcc acgccctcct acatcgaagc tgaaagcacg
     3541 agattcttcg ccctccgaga gctgcatcag gtcggagacg ctgtcgaact tttcgatcag
     3601 aaacttctcg acagacgtcg cggtgagttc aggctttttc atatcttatt gccccccggg
     3661 gccctcgact gtgttgatgc gggctgagat tggtggtggt ctatcacgaa tatgtgtgag
     3721 gggtaagtgc ggtgttttgc gtgagatttt agaatattgc cccgccccgg ggcaggccgg
     3781 cgtggcggaa caaccaggca cacgagcgcg aatggtgata ccgacggagt caaaactttg
     3841 tgacaagtag ctgcaccatg ggcagtggtg agctttcaga cgtggtatca ctgtccacta
     3901 gttcacacac agaatgcgtg tccaaaaggt ctagagccgt ctcgcttgcg tctctccgtc
     3961 gaagaacagt gaagaggctc gtcacgtcga ccagacgacg ggaggctggt caccatcgca
     4021 gatgtctccc acaaagcagc acggcaactc ctactccttc acacaatgga agaaaaggtg
     4081 gtctgatggt tctcagtgga aaagaacgat atcaggctga aaaaaatgat ctgcaggctc
     4141 cagattcctg aatcacgtcg actgtgacga agcaaaccgc gtcgaacaac atcggtcatg
     4201 ccaacgggtc tcgtctctcg agcccttttg gcggcgacta agaagtatga agcttcaggc
     4261 cgcaacgcgc gacacagcgt tttgtgtggt gggcctcggc attgctcttt gcatggccca
     4321 gcgtgattag tgcgtggatt ttaagcccga gaccgaagga ttgcgacatg tgcctggctg
     4381 tataactcac gcttgctgct acgctcgcct cctcctccat ccactccatc gcggccgcca
     4441 tggcctctag tggatcagct tgcatgcctg caggagacga tatcacctct tctgtttcca
     4501 cgataaaaat agactgctca tttcttcgtc gtcttcatcg tctgcttttt ctgcttcgcc
     4561 tctgtctggg gtctgaaacc actacacaca cacacaacac tcgtactccc actttcacaa
     4621 aagcgtaagc tcaccggctt ttcttacacg tacattttag tggatcccat cacgccacta
     4681 ccacgcccgc gggggatgga acggagggga gagagagaag ggggaagcat ggatgaatga
     4741 gacattgagg gaaaggaggg gagggagcag tccatcaggg cgctacctct cttgtccccc
     4801 aaaccctgtt gagccgttca acatgtttca tgtttcctcc ctcccccctt ccctccctgg
     4861 cctttccgcg gagccattca agtgacgtct ggaccgcacc gtaacaaaat cgtttctatg
     4921 gggggtttgt ttgacaacca cgtcttcagc gtttttaaaa aaaaaagcgg gcaagccctc
     4981 tcaccctcac tcatgcccat cctcctcctc tcctgcggaa cattcttaca aaaggcgtaa
     5041 ctcgacgaca actcaaagaa cgacaaacat caatcccaaa aaaaaaatct ctactcgtct
     5101 ctcttggatc tttccaattg tcagaccttc ctcttctttt tcgggctagc caccatggac
     5161 gcgtagtcgg gcacgtcgta ggggtatccg gccaggatgc gctcgcacag gcgccagccg
     5221 gttaccccgt tgatggtcac gcggaacaac agcgagccat ccgggttgat cagccgctcg
     5281 tcaatgatct tattgccgtt ccacagggtg ccggtcaccg tgatcttctt gccgtcgaac
     5341 acggcaatgc cctcgtaagg acggccgaag tagtcaatca tgttcggcgt cacaccgtcg
     5401 atgaccagcg tcccgtagtg cagtatcacc ttgaagtggt gatcgtccac ggggtacaca
     5461 accttgaaga tcttctcgat ctggcccatt tggtccccgc tcaggccctc gtagggaatg
     5521 atgacgtgga tgtcgatctt caggccgttc tcgccgctca ggacgatgcg ctggattggg
     5581 gtcacggaga cgcccaggtt ctggaaaagc gaggacacac cgccctgctc cagcacctgg
     5641 tcgaggttat agccagctgt ctgccgccag tcgcccacga agtcctcgag agtaaacacc
     5701 atgttaacgg acccgctgcc ggacccgtca gccctgctgt ctccaccgag ctgagagagg
     5761 tcgattcttg tttcatagag ccccgtaatt gactgatgaa tcagtgtggc gtccaggacc
     5821 tcctttgtag aggtgtaccg ctttctgtct atggtggtgt cgaagtactt gaaggctgca
     5881 ggcgcgccca agttggtcag agtaaacaag tggataatgt tttctgcctg ctccctgatg
     5941 ggcttatccc tgtgcttatt gtaagcagaa agcaccttat cgaggttagc gtcggcgagg
     6001 atcactcttt tggagaattc gcttatttgc tcgatgatct catcaaggta gtgtttgtgt
     6061 tgttccacga acagctgctt ctgctcatta tcttcgggag accctttgag cttttcatag
     6121 tggctggcca gatacaagaa attaacgtat ttagagggca gtgccagctc gttacctttc
     6181 tgcagctcgc ccgcactagc gagcattcgt ttccggccgt tttcaagctc aaagagagag
     6241 tacttgggaa gcttaatgat gaggtctttt ttgacctctt tatatccttt cgcctcgaga
     6301 aagtcgatgg ggtttttttc gaagcttgat cgctccatga ttgtgatgcc cagcagttcc
     6361 ttgacgcttt tgagtttttt agacttccct ttctccactt tggccacaac cagtacactg
     6421 taagcgactg taggagaatc gaatccgccg tatttcttgg ggtcccaatc ttttttgcgt
     6481 gcgatcagct tgtcgctgtt ccttttcggg aggatacttt ccttggagaa gcctccggtc
     6541 tgtacttcgg tctttttaac gatgttcacc tgcggcatgg acaggacctt ccggactgtc
     6601 gcgaaatccc tacccttgtc ccacacgatt tctcctgttt ctccgtttgt ttcgataagt
     6661 ggtcgcttcc gaatctctcc attggccagt gtaatctcgg tcttgaaaaa attcataata
     6721 ttgctgtaaa agaagtactt agcggtggcc ttgcctattt cctgctcaga ctttgcgatc
     6781 attttcctaa catcgtacac tttatagtct ccgtaaacaa attcagattc aagcttggga
     6841 tattttttga taagtgcagt gcctaccact gcattcaggt aggcatcatg cgcatggtgg
     6901 taattgttga tctctctcac cttataaaac tgaaagtcct ttctgaaatc tgagaccagc
     6961 ttagacttca gagtaataac tttcacctct cgaatcagtt tgtcattttc atcgtacttg
     7021 gtgttcatgc gtgaatcgag aatttgggcc acgtgcttgg tgatctggcg tgtctcaaca
     7081 agctgccttt tgatgaagcc ggctttatcc aactcagaca ggccacctcg ttcagcctta
     7141 gtcagattat cgaacttccg ttgtgtgatc agtttggcgt tcagcagctg ccgccaataa
     7201 tttttcattt tcttgacaac ttcttctgag gggacgttat cactcttccc tctattttta
     7261 tcggatcttg tcaacacttt attatcaata gaatcatctt tgagaaaaga ctggggcacg
     7321 atatgatcca cgtcgtagtc ggagagccga ttgatgtcca gttcctgatc cacgtacatg
     7381 tccctgccgt tctgcaggta gtacaggtag agcttctcat tctgaagctg ggtgttttca
     7441 actgggtgtt ccttaaggat ttgggacccc agttctttta taccctcttc aatcctcttc
     7501 atcctttccc tactgttctt ctgtcccttc tgggtagttt ggttctctcg ggccatctcg
     7561 ataacgatat tctcgggctt atgccttccc attactttga cgagttcatc cacgacctta
     7621 acggtctgca gtattccctt tttgatagct gggctacctg caagattagc gatgtgctcg
     7681 tgaagactgt ccccctggcc agaaacttgt gctttctgga tgtcctcctt aaaggtgaga
     7741 gagtcatcat ggatcaactg catgaagttc cggttggcaa atccatcgga cttaagaaaa
     7801 tccaggattg tctttccact ctgcttgtct cggatcccat tgatcagttt tcttgacagc
     7861 cgcccccatc ctgtatatcg gcgcctcttg agctgtttca tgactttgtc gtcgaagaga
     7921 tgagcgtaag ttttcaagcg ttcttcaatc atctccctat cttcaaacaa cgtaagggtg
     7981 aggacaatgt cctcaagaat gtcctcgttc tcctcattgt ccaggaagtc cttgtcttta
     8041 atgattttca ggagatcgtg atacgttccc agggatgcgt tgaagcgatc ctccactccg
     8101 ctgatttcaa cagagtcgaa acattcaatc tttttgaaat agtcttcttt gagctgtttc
     8161 acggtaactt tccggttcgt cttgaagagg aggtccacga tagctttctt ctgctctcca
     8221 gacaggaatg ctggctttct catcccttct gtgacgtatt tgaccttggt gagctcgtta
     8281 taaactgtga agtactcgta cagcagagag tgtttaggaa gcaccttttc gttaggcaga
     8341 tttttatcaa agttagtcat cctttcgatg aaggactggg cagaggcccc cttatccacg
     8401 acttcctcga agttccaggg agtgatggtc tcttctgatt tgcgagtcat ccacgcgaat
     8461 ctggaatttc cccgggcgag ggggcctaca tagtagggta tccgaaatgt gaggattttc
     8521 tcaatctttt ccctgttatc tttcaaaaag gggtagaaat cctcttgccg cctgaggata
     8581 gcgtgcagtt cgcccaggtg aatctggtgg gggatgcttc cattgtcgaa agtgcgctgt
     8641 ttgcgcaaca gatcttctct gttaagcttt accagcagct cctcggtgcc gtccattttt
     8701 tccaagatgg gcttaataaa tttgtaaaat tcctcctggc ttgctccgcc gtcaatgtat
     8761 ccggcgtagc catttttaga ctgatcgaag aaaatttcct tgtacttctc aggcagttgc
     8821 tgtctgacaa gggccttcag caaagtcaag tcttggtggt gctcatcata gcgcttgatc
     8881 atactagcgc tcagcggagc tttggtgatc tccgtgttca ctcgcagaat atcactcagc
     8941 agaatggcgt ctgacaggtt ctttgccgcc aaaaaaaggt ctgcgtactg gtcgccgatc
     9001 tgggccagca gattgtcgag atcatcatcg taggtgtctt tgctcagttg aagcttggca
     9061 tcttcggcca ggtcgaagtt agatttaaag ttgggggtca gcccgagtga cagggcgata
     9121 agattaccaa acaggccgtt cttcttctcc ccagggagct gtgcgatgag gttttcgagc
     9181 cgccgggatt tggacagcct agcgctcagg attgctttgg cgtcaactcc ggatgcgttg
     9241 atcgggttct cttcgaaaag ctgattgtaa gtctgaacca gttggataaa gagtttgtcg
     9301 acatcgctgt tgtctgggtt caggtccccc tcgatgagga agtgtccccg aaatttgatc
     9361 atatgcgcca gcgcgagata gatcaaccgc aagtcagcct tatcagtact gtctacaagc
     9421 ttcttcctca gatgatatat ggttgggtac ttttcatggt acgccacctc gtccacgata
     9481 ttgccaaaga ttgggtggcg ctcgtgcttt ttatcctcct ccaccaaaaa ggactcctcc
     9541 agcctatgga agaaagagtc atccacctta gccatctcat tactaaagat ctcctgcagg
     9601 tagcagatcc gattctttct gcgggtatat ctgcgccgtg ctgttctttt gagccgcgtg
     9661 gcttcggccg tctccccgga gtcgaacagg agggcgccaa tgaggttctt ctttatgctg
     9721 tggcgatcgg tattgcccag aactttgaat tttttgctcg gcaccttgta ctcgtccgta
     9781 atgacggccc agccgacgct gtttgtgccg atatcgagcc caatggagta cttcttgtcc
     9841 accttccgct ttttcttggg catgctcgag ggttgcgtgt gtatctgtgt gcagtggata
     9901 ttgttaccga gtttggtgag cgtgagtccg ttagtgccct ggtggtggtg gattaggaga
     9961 gtgggtgact cggtgtccat ggctttcttc gctcattata ggaggggaaa ggaatgaggg
    10021 agggtgggga gaccgcgtct gttgttgacc accgatttac ttcttgcctc ccttcccctc
    10081 cctcccctca atccgtacga cacaaatagt agccgagtgt ctgctgcaga gcgcatgatt
    10141 agtgtggtag acaacgaggg agggaaggat gtacagggca tggcacggag aagcgatggt
    10201 ggccaggaag aggagaggtc gcgagaacag gatgtgttgc gaatggataa aaacagaaag
    10261 cgatggctct gggcttcgaa agcaggggac attaggacgt gtagaccatc tcgacggatc
    10321 cctctgtatc tctgttgtgc gtgaatgttt tctgtgcacg tgtagtgtgt gagagtagaa
    10381 cccgggaact cgaacagaga aaagcatggg tggctgtggt gtggaggctt cgttcccacc
    10441 acatgccctt ctccttcgcc tcgcctctcc ctgccttctt ccacgcaccc ttgcgcccct
    10501 cgttctcaat acctggctca cttccaccat tcaaacaacc atcacgatac aggcatttat
    10561 ctatcgttga agacttcttc ctccggtaga tcttagccaa ggtaagaaga ggggcatgca
    10621 gcaaggagaa agaaatgatg catgatgagg aatagaaggg gaggagggag ggatatgatg
    10681 ggaagcgaaa gcgcatattc tggtggtctg ctgcctgatg gggacgcgtc tagctgtgac
    10741 actgaggacg gtggctgctg gtggctgcgg gcgctgcctt ggtgatcaat gggagtaaag
    10801 ggagggaagg aggtagcgtg aacggatgac gcggagaagt ttaggggtct ctttacgtat
    10861 cgcccctgcc gcccgcctct ctgcgataaa tgtgcctgtt accctgcagc ctctattctt
    10921 cactgtgttc ctgttttcca acagcctcta ttcttccctg tcttttgttg cagtggcgtc
    10981 atcctctctt tgccccagtc gtcgttctct cgactcactc actccccccc tccttccctc
    11041 cctccatcca cagaatcgat gaagagcagg ccttggctct tcgttttaga gctagaaata
    11101 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
    11161 ttggccggca tggtcccagc ctcctcgctg gcgccggctg ggcaacatgc ttcggcatgg
    11221 cgaatgggac tcctgggtac catgggaaag aaaggatgag aaaggagaaa ggacatctag
    11281 ataaccggca tatcacggtg gtgtattagt gtagaatagt gaagagaaga cttgggaaaa
    11341 tgtgtaggaa aggttgtttc tgtgtatgtg ggttgggatg ggtggctgtt tgagaaggaa
    11401 cagcgggcag ggcgatgtag tgctgaacgg gcacggaacc actgagactg aaggaagtag
    11461 ggagagagag gggcagggga cgtgcacttt aatctttgcc tcggtagagt atacccatgc
    11521 aagagtatgt ggccacctgt ggtggctttt ggccaggtct ggtgcagtgc caatcatctc
    11581 ccatcaataa tacaacttca gaacaacggc gcattgatgg ggagagagaa agtaaattta
    11641 agtaaggggt acgtagtaga ggattcaact gaaatatttt cgaggagcgg ttgggaagtt
    11701 gaccgattga aaggagaagg gaggggagca ggtgtgatag tcatgtgtaa agtaattctt
    11761 ttttgccgtc gtcacacaat ccacatcaat gataaaatat gtttaaggat caatcacacg
    11821 gagtcggtca taaggcaacc gcaaacgcaa tgcaaactag caagcaagca ggaccacaac
    11881 aacaaccatt accatcacag gcgacagcag cagcagctgc agcagcagca aggcaagcaa
    11941 aggccatctg cgcgtggact tttccaggca ccggctttaa gcgtaatttt caatgattgt
    12001 tgtccgtgta tcctctcacc ctttcaccgc ttgcgcgcac gtgagcagca caactggtaa
    12061 tgcccgagga caatagacga ggaaaaagaa gaagaaaaat gaagagggag tgatgaagaa
    12121 gaagagaaga ataaaagaag acagaaaaga agataaaaaa gtcaaaagac gacaaagacg
    12181 gaaaaaaaag aaacgcgcaa ttttaacacc agcaacgacg acgaaaaggg cgtcagcttt
    12241 gc
//
