LOCUS       sequence-192784-       12209 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 12209)
  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 12209)
  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..12209
                     /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(2586..3611)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(2586..3611)
                     /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      4401..4410
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(5091..5111)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      5116..5125
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(5127..5153)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(5706..9806)
                     /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(5706..9806)
                     /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(5706..9806)
                     /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(9807..9827)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(10048..10059)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_RNA        11050..11125
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     ncRNA           11130..11197
                     /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 gtttaaacag tagatgccga ccggagtccg caaaaatcac cagtctctct ctacaaatct
     2401 atctctctct atttttctcc agaataatgt gtgagtagtt cccagataag ggaattaggg
     2461 ttcttatagg gtttcgctca tgtgttgagc atataagaaa cccttagtat gtatttgtat
     2521 ttgtaaaata cttctatcaa taaaatttct aattcctaaa accaaaatcc agtgacctga
     2581 ctagcctatt cctttgccct cggacgagtg ctggggcgtc ggtttccact atcggcgagt
     2641 acttctacac agccatcggt ccagacggcc gcgcttctgc gggcgatttg tgtacgcccg
     2701 acagtcccgg ctccggatcg gacgattgcg tcgcatcgac cctgcgccca agctgcatca
     2761 tcgaaattgc cgtcaaccaa gctctgatag agttggtcaa gaccaatgcg gagcatatac
     2821 gcccggagcc gcggcgatcc tgcaagctcc ggatgcctcc gctcgaagta gcgcgtctgc
     2881 tgctccatac aagccaacca cggcctccag aagaagatgt tggcgacctc gtattgggaa
     2941 tccccgaaca tcgcctcgct ccagtcaatg accgctgtta tgcggccatt gtccgtcagg
     3001 acattgttgg agccgaaatc cgcgtgcacg aggtgccgga cttcggggca gtcctcggcc
     3061 caaagcatca gctcatcgag agcctgcgcg acggacgcac tgacggtgtc gtccatcaca
     3121 gtttgccagt gatacacatg gggatcagca atcgcgcata tgaaatcacg ccatgtagtg
     3181 tattgaccga ttccttgcgg tccgaatggg ccgaacccgc tcgtctggct aagatcggcc
     3241 gcagcgatcg catccatggc ctccgcgacc ggctgcagaa cagcgggcag ttcggtttca
     3301 ggcaggtctt gcaacgtgac accctgtgca cggcgggaga tgcaataggt caggctctcg
     3361 ctgaattccc caatgtcaag cacttccgga atcgggagcg cggccgatgc aaagtgccga
     3421 taaacataac gatctttgta gaaaccatcg gcgcagctat ttacccgcag gacatatcca
     3481 cgccctccta catcgaagct gaaagcacga gattcttcgc cctccgagag ctgcatcagg
     3541 tcggagacgc tgtcgaactt ttcgatcaga aacttctcga cagacgtcgc ggtgagttca
     3601 ggctttttca tatcttattg ccccccgggg ccctcctgtg ttgatgcggg ctgagattgg
     3661 tggtggtcta tcacgaatat gtgtgagggg taagtgcggt gttttgcgtg agattttaga
     3721 atattgcccc gccccggggc aggccggcgt ggcggaacaa ccaggcacac gagcgcgaat
     3781 ggtgataccg acggagtcaa aactttgtga caagtagctg caccatgggc agtggtgagc
     3841 tttcagacgt ggtatcactg tccactagtt cacacacaga atgcgtgtcc aaaaggtcta
     3901 gagccgtctc gcttgcgtct ctccgtcgaa gaacagtgaa gaggctcgtc acgtcgacca
     3961 gacgacggga ggctggtcac catcgcagat gtctcccaca aagcagcacg gcaactccta
     4021 ctccttcaca caatggaaga aaaggtggtc tgatggttct cagtggaaaa gaacgatatc
     4081 aggctgaaaa aaatgatctg caggctccag attcctgaat cacgtcgact gtgacgaagc
     4141 aaaccgcgtc gaacaacatc ggtcatgcca acgggtctcg tctctcgagc ccttttggcg
     4201 gcgactaaga agtatgaagc ttcaggccgc aacgcgcgac acagcgtttt gtgtggtggg
     4261 cctcggcatt gctctttgca tggcccagcg tgattagtgc gtggatttta agcccgagac
     4321 cgaaggattg cgacatgtgc ctggctgtat aactcacgct tgctgctacg ctcgcctcct
     4381 cctccatcca ctccatcgcg gccgccatgg cctctagtgg atcagcttgc atgcctgcag
     4441 gagacgatat cacctcttct gtttccacga taaaaataga ctgctcattt cttcgtcgtc
     4501 ttcatcgtct gctttttctg cttcgcctct gtctggggtc tgaaaccact acacacacac
     4561 acaacactcg tactcccact ttcacaaaag cgtaagctca ccggcttttc ttacacgtac
     4621 attttagtgg atcccatcac gccactacca cgcccgcggg ggatggaacg gaggggagag
     4681 agagaagggg gaagcatgga tgaatgagac attgagggaa aggaggggag ggagcagtcc
     4741 atcagggcgc tacctctctt gtcccccaaa ccctgttgag ccgttcaaca tgtttcatgt
     4801 ttcctccctc cccccttccc tccctggcct ttccgcggag ccattcaagt gacgtctgga
     4861 ccgcaccgta acaaaatcgt ttctatgggg ggtttgtttg acaaccacgt cttcagcgtt
     4921 tttaaaaaaa aaagcgggca agccctctca ccctcactca tgcccatcct cctcctctcc
     4981 tgcggaacat tcttacaaaa ggcgtaactc gacgacaact caaagaacga caaacatcaa
     5041 tcccaaaaaa aaaatctcta ctcgtctctc ttggatcttt ccaattgtca gaccttcctc
     5101 ttctttttcg ggctagccac catggacgcg tagtcgggca cgtcgtaggg gtatccggcc
     5161 aggatgcgct cgcacaggcg ccagccggtt accccgttga tggtcacgcg gaacaacagc
     5221 gagccatccg ggttgatcag ccgctcgtca atgatcttat tgccgttcca cagggtgccg
     5281 gtcaccgtga tcttcttgcc gtcgaacacg gcaatgccct cgtaaggacg gccgaagtag
     5341 tcaatcatgt tcggcgtcac accgtcgatg accagcgtcc cgtagtgcag tatcaccttg
     5401 aagtggtgat cgtccacggg gtacacaacc ttgaagatct tctcgatctg gcccatttgg
     5461 tccccgctca ggccctcgta gggaatgatg acgtggatgt cgatcttcag gccgttctcg
     5521 ccgctcagga cgatgcgctg gattggggtc acggagacgc ccaggttctg gaaaagcgag
     5581 gacacaccgc cctgctccag cacctggtcg aggttatagc cagctgtctg ccgccagtcg
     5641 cccacgaagt cctcgagagt aaacaccatg ttaacggacc cgctgccgga cccgtcagcc
     5701 ctgctgtctc caccgagctg agagaggtcg attcttgttt catagagccc cgtaattgac
     5761 tgatgaatca gtgtggcgtc caggacctcc tttgtagagg tgtaccgctt tctgtctatg
     5821 gtggtgtcga agtacttgaa ggctgcaggc gcgcccaagt tggtcagagt aaacaagtgg
     5881 ataatgtttt ctgcctgctc cctgatgggc ttatccctgt gcttattgta agcagaaagc
     5941 accttatcga ggttagcgtc ggcgaggatc actcttttgg agaattcgct tatttgctcg
     6001 atgatctcat caaggtagtg tttgtgttgt tccacgaaca gctgcttctg ctcattatct
     6061 tcgggagacc ctttgagctt ttcatagtgg ctggccagat acaagaaatt aacgtattta
     6121 gagggcagtg ccagctcgtt acctttctgc agctcgcccg cactagcgag cattcgtttc
     6181 cggccgtttt caagctcaaa gagagagtac ttgggaagct taatgatgag gtcttttttg
     6241 acctctttat atcctttcgc ctcgagaaag tcgatggggt ttttttcgaa gcttgatcgc
     6301 tccatgattg tgatgcccag cagttccttg acgcttttga gttttttaga cttccctttc
     6361 tccactttgg ccacaaccag tacactgtaa gcgactgtag gagaatcgaa tccgccgtat
     6421 ttcttggggt cccaatcttt tttgcgtgcg atcagcttgt cgctgttcct tttcgggagg
     6481 atactttcct tggagaagcc tccggtctgt acttcggtct ttttaacgat gttcacctgc
     6541 ggcatggaca ggaccttccg gactgtcgcg aaatccctac ccttgtccca cacgatttct
     6601 cctgtttctc cgtttgtttc gataagtggt cgcttccgaa tctctccatt ggccagtgta
     6661 atctcggtct tgaaaaaatt cataatattg ctgtaaaaga agtacttagc ggtggccttg
     6721 cctatttcct gctcagactt tgcgatcatt ttcctaacat cgtacacttt atagtctccg
     6781 taaacaaatt cagattcaag cttgggatat tttttgataa gtgcagtgcc taccactgca
     6841 ttcaggtagg catcatgcgc atggtggtaa ttgttgatct ctctcacctt ataaaactga
     6901 aagtcctttc tgaaatctga gaccagctta gacttcagag taataacttt cacctctcga
     6961 atcagtttgt cattttcatc gtacttggtg ttcatgcgtg aatcgagaat ttgggccacg
     7021 tgcttggtga tctggcgtgt ctcaacaagc tgccttttga tgaagccggc tttatccaac
     7081 tcagacaggc cacctcgttc agccttagtc agattatcga acttccgttg tgtgatcagt
     7141 ttggcgttca gcagctgccg ccaataattt ttcattttct tgacaacttc ttctgagggg
     7201 acgttatcac tcttccctct atttttatcg gatcttgtca acactttatt atcaatagaa
     7261 tcatctttga gaaaagactg gggcacgata tgatccacgt cgtagtcgga gagccgattg
     7321 atgtccagtt cctgatccac gtacatgtcc ctgccgttct gcaggtagta caggtagagc
     7381 ttctcattct gaagctgggt gttttcaact gggtgttcct taaggatttg ggaccccagt
     7441 tcttttatac cctcttcaat cctcttcatc ctttccctac tgttcttctg tcccttctgg
     7501 gtagtttggt tctctcgggc catctcgata acgatattct cgggcttatg ccttcccatt
     7561 actttgacga gttcatccac gaccttaacg gtctgcagta ttcccttttt gatagctggg
     7621 ctacctgcaa gattagcgat gtgctcgtga agactgtccc cctggccaga aacttgtgct
     7681 ttctggatgt cctccttaaa ggtgagagag tcatcatgga tcaactgcat gaagttccgg
     7741 ttggcaaatc catcggactt aagaaaatcc aggattgtct ttccactctg cttgtctcgg
     7801 atcccattga tcagttttct tgacagccgc ccccatcctg tatatcggcg cctcttgagc
     7861 tgtttcatga ctttgtcgtc gaagagatga gcgtaagttt tcaagcgttc ttcaatcatc
     7921 tccctatctt caaacaacgt aagggtgagg acaatgtcct caagaatgtc ctcgttctcc
     7981 tcattgtcca ggaagtcctt gtctttaatg attttcagga gatcgtgata cgttcccagg
     8041 gatgcgttga agcgatcctc cactccgctg atttcaacag agtcgaaaca ttcaatcttt
     8101 ttgaaatagt cttctttgag ctgtttcacg gtaactttcc ggttcgtctt gaagaggagg
     8161 tccacgatag ctttcttctg ctctccagac aggaatgctg gctttctcat cccttctgtg
     8221 acgtatttga ccttggtgag ctcgttataa actgtgaagt actcgtacag cagagagtgt
     8281 ttaggaagca ccttttcgtt aggcagattt ttatcaaagt tagtcatcct ttcgatgaag
     8341 gactgggcag aggccccctt atccacgact tcctcgaagt tccagggagt gatggtctct
     8401 tctgatttgc gagtcatcca cgcgaatctg gaatttcccc gggcgagggg gcctacatag
     8461 tagggtatcc gaaatgtgag gattttctca atcttttccc tgttatcttt caaaaagggg
     8521 tagaaatcct cttgccgcct gaggatagcg tgcagttcgc ccaggtgaat ctggtggggg
     8581 atgcttccat tgtcgaaagt gcgctgtttg cgcaacagat cttctctgtt aagctttacc
     8641 agcagctcct cggtgccgtc cattttttcc aagatgggct taataaattt gtaaaattcc
     8701 tcctggcttg ctccgccgtc aatgtatccg gcgtagccat ttttagactg atcgaagaaa
     8761 atttccttgt acttctcagg cagttgctgt ctgacaaggg ccttcagcaa agtcaagtct
     8821 tggtggtgct catcatagcg cttgatcata ctagcgctca gcggagcttt ggtgatctcc
     8881 gtgttcactc gcagaatatc actcagcaga atggcgtctg acaggttctt tgccgccaaa
     8941 aaaaggtctg cgtactggtc gccgatctgg gccagcagat tgtcgagatc atcatcgtag
     9001 gtgtctttgc tcagttgaag cttggcatct tcggccaggt cgaagttaga tttaaagttg
     9061 ggggtcagcc cgagtgacag ggcgataaga ttaccaaaca ggccgttctt cttctcccca
     9121 gggagctgtg cgatgaggtt ttcgagccgc cgggatttgg acagcctagc gctcaggatt
     9181 gctttggcgt caactccgga tgcgttgatc gggttctctt cgaaaagctg attgtaagtc
     9241 tgaaccagtt ggataaagag tttgtcgaca tcgctgttgt ctgggttcag gtccccctcg
     9301 atgaggaagt gtccccgaaa tttgatcata tgcgccagcg cgagatagat caaccgcaag
     9361 tcagccttat cagtactgtc tacaagcttc ttcctcagat gatatatggt tgggtacttt
     9421 tcatggtacg ccacctcgtc cacgatattg ccaaagattg ggtggcgctc gtgcttttta
     9481 tcctcctcca ccaaaaagga ctcctccagc ctatggaaga aagagtcatc caccttagcc
     9541 atctcattac taaagatctc ctgcaggtag cagatccgat tctttctgcg ggtatatctg
     9601 cgccgtgctg ttcttttgag ccgcgtggct tcggccgtct ccccggagtc gaacaggagg
     9661 gcgccaatga ggttcttctt tatgctgtgg cgatcggtat tgcccagaac tttgaatttt
     9721 ttgctcggca ccttgtactc gtccgtaatg acggcccagc cgacgctgtt tgtgccgata
     9781 tcgagcccaa tggagtactt cttgtccacc ttccgctttt tcttgggcat gctcgagggt
     9841 tgcgtgtgta tctgtgtgca gtggatattg ttaccgagtt tggtgagcgt gagtccgtta
     9901 gtgccctggt ggtggtggat taggagagtg ggtgactcgg tgtccatggc tttcttcgct
     9961 cattatagga ggggaaagga atgagggagg gtggggagac cgcgtctgtt gttgaccacc
    10021 gatttacttc ttgcctccct tcccctccct cccctcaatc cgtacgacac aaatagtagc
    10081 cgagtgtctg ctgcagagcg catgattagt gtggtagaca acgagggagg gaaggatgta
    10141 cagggcatgg cacggagaag cgatggtggc caggaagagg agaggtcgcg agaacaggat
    10201 gtgttgcgaa tggataaaaa cagaaagcga tggctctggg cttcgaaagc aggggacatt
    10261 aggacgtgta gaccatctcg acggatccct ctgtatctct gttgtgcgtg aatgttttct
    10321 gtgcacgtgt agtgtgtgag agtagaaccc gggaactcga acagagaaaa gcatgggtgg
    10381 ctgtggtgtg gaggcttcgt tcccaccaca tgcccttctc cttcgcctcg cctctccctg
    10441 ccttcttcca cgcacccttg cgcccctcgt tctcaatacc tggctcactt ccaccattca
    10501 aacaaccatc acgatacagg catttatcta tcgttgaaga cttcttcctc cggtagatct
    10561 tagccaaggt aagaagaggg gcatgcagca aggagaaaga aatgatgcat gatgaggaat
    10621 agaaggggag gagggaggga tatgatggga agcgaaagcg catattctgg tggtctgctg
    10681 cctgatgggg acgcgtctag ctgtgacact gaggacggtg gctgctggtg gctgcgggcg
    10741 ctgccttggt gatcaatggg agtaaaggga gggaaggagg tagcgtgaac ggatgacgcg
    10801 gagaagttta ggggtctctt tacgtatcgc ccctgccgcc cgcctctctg cgataaatgt
    10861 gcctgttacc ctgcagcctc tattcttcac tgtgttcctg ttttccaaca gcctctattc
    10921 ttccctgtct tttgttgcag tggcgtcatc ctctctttgc cccagtcgtc gttctctcga
    10981 ctcactcact ccccccctcc ttccctccct ccatccacag aatcgatgaa gagcaggcct
    11041 tggctcttcg ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact
    11101 tgaaaaagtg gcaccgagtc ggtgcttttg gccggcatgg tcccagcctc ctcgctggcg
    11161 ccggctgggc aacatgcttc ggcatggcga atgggactcc tgggtaccat gggaaagaaa
    11221 ggatgagaaa ggagaaagga catctagata accggcatat cacggtggtg tattagtgta
    11281 gaatagtgaa gagaagactt gggaaaatgt gtaggaaagg ttgtttctgt gtatgtgggt
    11341 tgggatgggt ggctgtttga gaaggaacag cgggcagggc gatgtagtgc tgaacgggca
    11401 cggaaccact gagactgaag gaagtaggga gagagagggg caggggacgt gcactttaat
    11461 ctttgcctcg gtagagtata cccatgcaag agtatgtggc cacctgtggt ggcttttggc
    11521 caggtctggt gcagtgccaa tcatctccca tcaataatac aacttcagaa caacggcgca
    11581 ttgatgggga gagagaaagt aaatttaagt aaggggtacg tagtagagga ttcaactgaa
    11641 atattttcga ggagcggttg ggaagttgac cgattgaaag gagaagggag gggagcaggt
    11701 gtgatagtca tgtgtaaagt aattcttttt tgccgtcgtc acacaatcca catcaatgat
    11761 aaaatatgtt taaggatcaa tcacacggag tcggtcataa ggcaaccgca aacgcaatgc
    11821 aaactagcaa gcaagcagga ccacaacaac aaccattacc atcacaggcg acagcagcag
    11881 cagctgcagc agcagcaagg caagcaaagg ccatctgcgc gtggactttt ccaggcaccg
    11941 gctttaagcg taattttcaa tgattgttgt ccgtgtatcc tctcaccctt tcaccgcttg
    12001 cgcgcacgtg agcagcacaa ctggtaatgc ccgaggacaa tagacgagga aaaagaagaa
    12061 gaaaaatgaa gagggagtga tgaagaagaa gagaagaata aaagaagaca gaaaagaaga
    12121 taaaaaagtc aaaagacgac aaagacggaa aaaaaagaaa cgcgcaattt taacaccagc
    12181 aacgacgacg aaaagggcgt cagctttgc
//
