LOCUS       sequence-202008-       11536 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cas9-Nlux and sgRNA and contains CEN/ARS. CCMP537 LDSP 
            promoter driven Blasticidin resistance gene..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11536)
  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 11536)
  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: pNSA-ARS-CRISPR-BlastR
FEATURES             Location/Qualifiers
     source          1..11536
                     /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"
     regulatory      3728..3737
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(4418..4438)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      4443..4452
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(4454..4480)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(5033..9133)
                     /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(5033..9133)
                     /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(5033..9133)
                     /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(9134..9154)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(9375..9386)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_RNA        10377..10452
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     ncRNA           10457..10524
                     /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 actagttcaa gcgctctccc acacgtagcc
     2641 cgaagggaga agttctctga taccgaccgc cgtaggctgc ccgtcggaat ccttcacaat
     2701 cgccttgatc ccagggtgga ggtcgagcag gacctgccgg caccgcccgc aggggctcag
     2761 aatcccgcgg ttctcgttcc cgatggcgac aatgcacgtc aggttcccag ccgcggcagc
     2821 agcggcggtc cccaggacca ccagctccgc gcaaggcccc cccgtaaaat ggtacacgtt
     2881 gacccccgtg aagatccgtc cgtcggacga cagagcagcg gaagccaccg agtaatcctc
     2941 ggagatagga atcgagttaa tcgtcgccgt agccctctcg atcagcgtgc tctcctcctg
     3001 cgagaggggc atgggcccct cgagcttatc ttgagataga atttgtgcta gtgtgtctac
     3061 gatgtgttac agtgcgagcg ccgtgagagg gtgcgtggca tcagattcca ttcctaattc
     3121 ggactgacgc cgtacacagg aaaacggcaa gcaacccaaa caaccacatc ctatcaaatg
     3181 caatacacga aatggtccag catgtcaaac acgtaaaaat cgttgtgtgg actgctcgta
     3241 gagaccgtag ccgtcaaaaa gacgcatatc acggtcaaat cttgtgagtt ctccaagaac
     3301 aaacggccag gttggctgtc gagtcgaagc acagaaagat gctcgtcaca agccgcctcc
     3361 ccatgtctga cgctaccaaa aacttggtat tccgtggtca agtccttgac ccgccccggc
     3421 agaaatgcag ggcaaaattt ataccagaca aggccactct gccgcgtacg gtggtcgtta
     3481 cgcacatggc ttcgtcacag ttgcccggtg ctttgtcgcc tatggcagcg aacatgacgc
     3541 aagcgggtct caacggccga cgagcgtctg cgcgagaaat gagcaaatag accgcaccca
     3601 cccatcacaa caacgcgcgc gggcgaataa atgcatggcc cggacgttct ggggtggggt
     3661 gctgttggca ggtgggaggg actgtattgt cttgaggatg gatgttgact tgatttggta
     3721 cacagcggcc gccatggcct ctagtggatc agcttgcatg cctgcaggag acgatatcac
     3781 ctcttctgtt tccacgataa aaatagactg ctcatttctt cgtcgtcttc atcgtctgct
     3841 ttttctgctt cgcctctgtc tggggtctga aaccactaca cacacacaca acactcgtac
     3901 tcccactttc acaaaagcgt aagctcaccg gcttttctta cacgtacatt ttagtggatc
     3961 ccatcacgcc actaccacgc ccgcggggga tggaacggag gggagagaga gaagggggaa
     4021 gcatggatga atgagacatt gagggaaagg aggggaggga gcagtccatc agggcgctac
     4081 ctctcttgtc ccccaaaccc tgttgagccg ttcaacatgt ttcatgtttc ctccctcccc
     4141 ccttccctcc ctggcctttc cgcggagcca ttcaagtgac gtctggaccg caccgtaaca
     4201 aaatcgtttc tatggggggt ttgtttgaca accacgtctt cagcgttttt aaaaaaaaaa
     4261 gcgggcaagc cctctcaccc tcactcatgc ccatcctcct cctctcctgc ggaacattct
     4321 tacaaaaggc gtaactcgac gacaactcaa agaacgacaa acatcaatcc caaaaaaaaa
     4381 atctctactc gtctctcttg gatctttcca attgtcagac cttcctcttc tttttcgggc
     4441 tagccaccat ggacgcgtag tcgggcacgt cgtaggggta tccggccagg atgcgctcgc
     4501 acaggcgcca gccggttacc ccgttgatgg tcacgcggaa caacagcgag ccatccgggt
     4561 tgatcagccg ctcgtcaatg atcttattgc cgttccacag ggtgccggtc accgtgatct
     4621 tcttgccgtc gaacacggca atgccctcgt aaggacggcc gaagtagtca atcatgttcg
     4681 gcgtcacacc gtcgatgacc agcgtcccgt agtgcagtat caccttgaag tggtgatcgt
     4741 ccacggggta cacaaccttg aagatcttct cgatctggcc catttggtcc ccgctcaggc
     4801 cctcgtaggg aatgatgacg tggatgtcga tcttcaggcc gttctcgccg ctcaggacga
     4861 tgcgctggat tggggtcacg gagacgccca ggttctggaa aagcgaggac acaccgccct
     4921 gctccagcac ctggtcgagg ttatagccag ctgtctgccg ccagtcgccc acgaagtcct
     4981 cgagagtaaa caccatgtta acggacccgc tgccggaccc gtcagccctg ctgtctccac
     5041 cgagctgaga gaggtcgatt cttgtttcat agagccccgt aattgactga tgaatcagtg
     5101 tggcgtccag gacctccttt gtagaggtgt accgctttct gtctatggtg gtgtcgaagt
     5161 acttgaaggc tgcaggcgcg cccaagttgg tcagagtaaa caagtggata atgttttctg
     5221 cctgctccct gatgggctta tccctgtgct tattgtaagc agaaagcacc ttatcgaggt
     5281 tagcgtcggc gaggatcact cttttggaga attcgcttat ttgctcgatg atctcatcaa
     5341 ggtagtgttt gtgttgttcc acgaacagct gcttctgctc attatcttcg ggagaccctt
     5401 tgagcttttc atagtggctg gccagataca agaaattaac gtatttagag ggcagtgcca
     5461 gctcgttacc tttctgcagc tcgcccgcac tagcgagcat tcgtttccgg ccgttttcaa
     5521 gctcaaagag agagtacttg ggaagcttaa tgatgaggtc ttttttgacc tctttatatc
     5581 ctttcgcctc gagaaagtcg atggggtttt tttcgaagct tgatcgctcc atgattgtga
     5641 tgcccagcag ttccttgacg cttttgagtt ttttagactt ccctttctcc actttggcca
     5701 caaccagtac actgtaagcg actgtaggag aatcgaatcc gccgtatttc ttggggtccc
     5761 aatctttttt gcgtgcgatc agcttgtcgc tgttcctttt cgggaggata ctttccttgg
     5821 agaagcctcc ggtctgtact tcggtctttt taacgatgtt cacctgcggc atggacagga
     5881 ccttccggac tgtcgcgaaa tccctaccct tgtcccacac gatttctcct gtttctccgt
     5941 ttgtttcgat aagtggtcgc ttccgaatct ctccattggc cagtgtaatc tcggtcttga
     6001 aaaaattcat aatattgctg taaaagaagt acttagcggt ggccttgcct atttcctgct
     6061 cagactttgc gatcattttc ctaacatcgt acactttata gtctccgtaa acaaattcag
     6121 attcaagctt gggatatttt ttgataagtg cagtgcctac cactgcattc aggtaggcat
     6181 catgcgcatg gtggtaattg ttgatctctc tcaccttata aaactgaaag tcctttctga
     6241 aatctgagac cagcttagac ttcagagtaa taactttcac ctctcgaatc agtttgtcat
     6301 tttcatcgta cttggtgttc atgcgtgaat cgagaatttg ggccacgtgc ttggtgatct
     6361 ggcgtgtctc aacaagctgc cttttgatga agccggcttt atccaactca gacaggccac
     6421 ctcgttcagc cttagtcaga ttatcgaact tccgttgtgt gatcagtttg gcgttcagca
     6481 gctgccgcca ataatttttc attttcttga caacttcttc tgaggggacg ttatcactct
     6541 tccctctatt tttatcggat cttgtcaaca ctttattatc aatagaatca tctttgagaa
     6601 aagactgggg cacgatatga tccacgtcgt agtcggagag ccgattgatg tccagttcct
     6661 gatccacgta catgtccctg ccgttctgca ggtagtacag gtagagcttc tcattctgaa
     6721 gctgggtgtt ttcaactggg tgttccttaa ggatttggga ccccagttct tttataccct
     6781 cttcaatcct cttcatcctt tccctactgt tcttctgtcc cttctgggta gtttggttct
     6841 ctcgggccat ctcgataacg atattctcgg gcttatgcct tcccattact ttgacgagtt
     6901 catccacgac cttaacggtc tgcagtattc cctttttgat agctgggcta cctgcaagat
     6961 tagcgatgtg ctcgtgaaga ctgtccccct ggccagaaac ttgtgctttc tggatgtcct
     7021 ccttaaaggt gagagagtca tcatggatca actgcatgaa gttccggttg gcaaatccat
     7081 cggacttaag aaaatccagg attgtctttc cactctgctt gtctcggatc ccattgatca
     7141 gttttcttga cagccgcccc catcctgtat atcggcgcct cttgagctgt ttcatgactt
     7201 tgtcgtcgaa gagatgagcg taagttttca agcgttcttc aatcatctcc ctatcttcaa
     7261 acaacgtaag ggtgaggaca atgtcctcaa gaatgtcctc gttctcctca ttgtccagga
     7321 agtccttgtc tttaatgatt ttcaggagat cgtgatacgt tcccagggat gcgttgaagc
     7381 gatcctccac tccgctgatt tcaacagagt cgaaacattc aatctttttg aaatagtctt
     7441 ctttgagctg tttcacggta actttccggt tcgtcttgaa gaggaggtcc acgatagctt
     7501 tcttctgctc tccagacagg aatgctggct ttctcatccc ttctgtgacg tatttgacct
     7561 tggtgagctc gttataaact gtgaagtact cgtacagcag agagtgttta ggaagcacct
     7621 tttcgttagg cagattttta tcaaagttag tcatcctttc gatgaaggac tgggcagagg
     7681 cccccttatc cacgacttcc tcgaagttcc agggagtgat ggtctcttct gatttgcgag
     7741 tcatccacgc gaatctggaa tttccccggg cgagggggcc tacatagtag ggtatccgaa
     7801 atgtgaggat tttctcaatc ttttccctgt tatctttcaa aaaggggtag aaatcctctt
     7861 gccgcctgag gatagcgtgc agttcgccca ggtgaatctg gtgggggatg cttccattgt
     7921 cgaaagtgcg ctgtttgcgc aacagatctt ctctgttaag ctttaccagc agctcctcgg
     7981 tgccgtccat tttttccaag atgggcttaa taaatttgta aaattcctcc tggcttgctc
     8041 cgccgtcaat gtatccggcg tagccatttt tagactgatc gaagaaaatt tccttgtact
     8101 tctcaggcag ttgctgtctg acaagggcct tcagcaaagt caagtcttgg tggtgctcat
     8161 catagcgctt gatcatacta gcgctcagcg gagctttggt gatctccgtg ttcactcgca
     8221 gaatatcact cagcagaatg gcgtctgaca ggttctttgc cgccaaaaaa aggtctgcgt
     8281 actggtcgcc gatctgggcc agcagattgt cgagatcatc atcgtaggtg tctttgctca
     8341 gttgaagctt ggcatcttcg gccaggtcga agttagattt aaagttgggg gtcagcccga
     8401 gtgacagggc gataagatta ccaaacaggc cgttcttctt ctccccaggg agctgtgcga
     8461 tgaggttttc gagccgccgg gatttggaca gcctagcgct caggattgct ttggcgtcaa
     8521 ctccggatgc gttgatcggg ttctcttcga aaagctgatt gtaagtctga accagttgga
     8581 taaagagttt gtcgacatcg ctgttgtctg ggttcaggtc cccctcgatg aggaagtgtc
     8641 cccgaaattt gatcatatgc gccagcgcga gatagatcaa ccgcaagtca gccttatcag
     8701 tactgtctac aagcttcttc ctcagatgat atatggttgg gtacttttca tggtacgcca
     8761 cctcgtccac gatattgcca aagattgggt ggcgctcgtg ctttttatcc tcctccacca
     8821 aaaaggactc ctccagccta tggaagaaag agtcatccac cttagccatc tcattactaa
     8881 agatctcctg caggtagcag atccgattct ttctgcgggt atatctgcgc cgtgctgttc
     8941 ttttgagccg cgtggcttcg gccgtctccc cggagtcgaa caggagggcg ccaatgaggt
     9001 tcttctttat gctgtggcga tcggtattgc ccagaacttt gaattttttg ctcggcacct
     9061 tgtactcgtc cgtaatgacg gcccagccga cgctgtttgt gccgatatcg agcccaatgg
     9121 agtacttctt gtccaccttc cgctttttct tgggcatgct cgagggttgc gtgtgtatct
     9181 gtgtgcagtg gatattgtta ccgagtttgg tgagcgtgag tccgttagtg ccctggtggt
     9241 ggtggattag gagagtgggt gactcggtgt ccatggcttt cttcgctcat tataggaggg
     9301 gaaaggaatg agggagggtg gggagaccgc gtctgttgtt gaccaccgat ttacttcttg
     9361 cctcccttcc cctccctccc ctcaatccgt acgacacaaa tagtagccga gtgtctgctg
     9421 cagagcgcat gattagtgtg gtagacaacg agggagggaa ggatgtacag ggcatggcac
     9481 ggagaagcga tggtggccag gaagaggaga ggtcgcgaga acaggatgtg ttgcgaatgg
     9541 ataaaaacag aaagcgatgg ctctgggctt cgaaagcagg ggacattagg acgtgtagac
     9601 catctcgacg gatccctctg tatctctgtt gtgcgtgaat gttttctgtg cacgtgtagt
     9661 gtgtgagagt agaacccggg aactcgaaca gagaaaagca tgggtggctg tggtgtggag
     9721 gcttcgttcc caccacatgc ccttctcctt cgcctcgcct ctccctgcct tcttccacgc
     9781 acccttgcgc ccctcgttct caatacctgg ctcacttcca ccattcaaac aaccatcacg
     9841 atacaggcat ttatctatcg ttgaagactt cttcctccgg tagatcttag ccaaggtaag
     9901 aagaggggca tgcagcaagg agaaagaaat gatgcatgat gaggaataga aggggaggag
     9961 ggagggatat gatgggaagc gaaagcgcat attctggtgg tctgctgcct gatggggacg
    10021 cgtctagctg tgacactgag gacggtggct gctggtggct gcgggcgctg ccttggtgat
    10081 caatgggagt aaagggaggg aaggaggtag cgtgaacgga tgacgcggag aagtttaggg
    10141 gtctctttac gtatcgcccc tgccgcccgc ctctctgcga taaatgtgcc tgttaccctg
    10201 cagcctctat tcttcactgt gttcctgttt tccaacagcc tctattcttc cctgtctttt
    10261 gttgcagtgg cgtcatcctc tctttgcccc agtcgtcgtt ctctcgactc actcactccc
    10321 cccctccttc cctccctcca tccacagaat cgatgaagag caggccttgg ctcttcgttt
    10381 tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca
    10441 ccgagtcggt gcttttggcc ggcatggtcc cagcctcctc gctggcgccg gctgggcaac
    10501 atgcttcggc atggcgaatg ggactcctgg gtaccatggg aaagaaagga tgagaaagga
    10561 gaaaggacat ctagataacc ggcatatcac ggtggtgtat tagtgtagaa tagtgaagag
    10621 aagacttggg aaaatgtgta ggaaaggttg tttctgtgta tgtgggttgg gatgggtggc
    10681 tgtttgagaa ggaacagcgg gcagggcgat gtagtgctga acgggcacgg aaccactgag
    10741 actgaaggaa gtagggagag agaggggcag gggacgtgca ctttaatctt tgcctcggta
    10801 gagtataccc atgcaagagt atgtggccac ctgtggtggc ttttggccag gtctggtgca
    10861 gtgccaatca tctcccatca ataatacaac ttcagaacaa cggcgcattg atggggagag
    10921 agaaagtaaa tttaagtaag gggtacgtag tagaggattc aactgaaata ttttcgagga
    10981 gcggttggga agttgaccga ttgaaaggag aagggagggg agcaggtgtg atagtcatgt
    11041 gtaaagtaat tcttttttgc cgtcgtcaca caatccacat caatgataaa atatgtttaa
    11101 ggatcaatca cacggagtcg gtcataaggc aaccgcaaac gcaatgcaaa ctagcaagca
    11161 agcaggacca caacaacaac cattaccatc acaggcgaca gcagcagcag ctgcagcagc
    11221 agcaaggcaa gcaaaggcca tctgcgcgtg gacttttcca ggcaccggct ttaagcgtaa
    11281 ttttcaatga ttgttgtccg tgtatcctct caccctttca ccgcttgcgc gcacgtgagc
    11341 agcacaactg gtaatgcccg aggacaatag acgaggaaaa agaagaagaa aaatgaagag
    11401 ggagtgatga agaagaagag aagaataaaa gaagacagaa aagaagataa aaaagtcaaa
    11461 agacgacaaa gacggaaaaa aaagaaacgc gcaattttaa caccagcaac gacgacgaaa
    11521 agggcgtcag ctttgc
//
