LOCUS       sequence-347041-       11636 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pNOC episomal plasmid harboring the dead version of humanized spCas9
            gene sequence tagged with Nlux and sgRNA targeting the fluorescent 
            gene tdtomato with spacer 2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11636)
  AUTHORS   Naduthodi MIS, Sudfeld C, Avitzigiannis EK, Trevisan N, van Lith E, 
            Alcaide Sancho J, D'Adamo S, Barbosa M, van der Oost J
  TITLE     Comprehensive Genome Engineering Toolbox for Microalgae 
            Nannochloropsis oceanica Based on CRISPR-Cas Systems.
  JOURNAL   ACS Synth Biol. 2021 Dec 17;10(12):3369-3378. doi: 
            10.1021/acssynbio.1c00329. Epub 2021 Nov 18.
  PUBMED    34793143
REFERENCE   2  (bases 1 to 11636)
  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.1c00329"; journalName: "ACS Synth Biol"; date: 
            "2021-12-17- 17"; volume: "10"; issue: "12"; pages: "3369-3378"
COMMENT     Sequence Label: pNOC_dCas9_sgRNA Td2
FEATURES             Location/Qualifiers
     source          1..11636
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        114..218
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(219..287,288..1079)
                     /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(437..456)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1250..1838
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1739..1758
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4027..4047)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      4052..4061
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(4063..4089)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(join(4642..6156,6157..6159,6160..6225,6226..6231,
                     6232..8715,8716..8718,8719..8742))
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=Cas9m4
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D10A, D839A, 
                     H840A, and N863A (Mali et al., 2013)"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(join(4642..6225,6226..6228,6229..8715,8716..8718,
                     8719..8742))
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(4642..8742)
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(8743..8763)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(8984..8995)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     ncRNA           9977..10013
                     /product="self-cleaving hammerhead ribozyme (Ferre-D'Amare 
                     and Scott, 2010)"
                     /label=hammerhead ribozyme
                     /ncRNA_class="hammerhead_ribozyme"
     misc_RNA        10034..10109
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     ncRNA           10114..10181
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     misc_feature    join(11210..11636,1..77)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
ORIGIN
        1 ctttttttac tttctatttt taatttatat atttatatta aaaaatttaa attataatta
       61 tttttatagc acgtgatgaa aaggacccag gtggcacttt tcggggaaat gtgcgcggaa
      121 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
      181 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
      241 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
      301 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
      361 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
      421 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
      481 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
      541 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
      601 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
      661 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
      721 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
      781 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
      841 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
      901 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
      961 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     1021 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     1081 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     1141 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     1201 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     1261 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     1321 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     1381 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     1441 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     1501 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     1561 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     1621 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     1681 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     1741 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     1801 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     1861 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     1921 attctgtggt ttaaaccagg tcactggatt ttggttttag gaattagaaa ttttattgat
     1981 agaagtattt tacaaataca aatacatact aagggtttct tatatgctca acacatgagc
     2041 gaaaccctat aagaacccta attcccttat ctgggaacta ctcacacatt attctggaga
     2101 aaaatagaga gagatagatt tgtagagaga gactggtgat ttttgcggac tccggtcggc
     2161 atctactact agttcaagcg ctctcccaca cgtagcccga agggagaagt tctctgatac
     2221 cgaccgccgt aggctgcccg tcggaatcct tcacaatcgc cttgatccca gggtggaggt
     2281 cgagcaggac ctgccggcac cgcccgcagg ggctcagaat cccgcggttc tcgttcccga
     2341 tggcgacaat gcacgtcagg ttcccagccg cggcagcagc ggcggtcccc aggaccacca
     2401 gctccgcgca aggccccccc gtaaaatggt acacgttgac ccccgtgaag atccgtccgt
     2461 cggacgacag agcagcggaa gccaccgagt aatcctcgga gataggaatc gagttaatcg
     2521 tcgccgtagc cctctcgatc agcgtgctct cctcctgcga gaggggcatg ggccctcctg
     2581 tgttgatgcg ggctgagatt ggtggtggtc tatcacgaat atgtgtgagg ggtaagtgcg
     2641 gtgttttgcg tgagatttta gaatattgcc ccgccccggg gcaggccggc gtggcggaac
     2701 aaccaggcac acgagcgcga atggtgatac cgacggagtc aaaactttgt gacaagtagc
     2761 tgcaccatgg gcagtggtga gctttcagac gtggtatcac tgtccactag ttcacacaca
     2821 gaatgcgtgt ccaaaaggtc tagagccgtc tcgcttgcgt ctctccgtcg aagaacagtg
     2881 aagaggctcg tcacgtcgac cagacgacgg gaggctggtc accatcgcag atgtctccca
     2941 caaagcagca cggcaactcc tactccttca cacaatggaa gaaaaggtgg tctgatggtt
     3001 ctcagtggaa aagaacgata tcaggctgaa aaaaatgatc tgcaggctcc agattcctga
     3061 atcacgtcga ctgtgacgaa gcaaaccgcg tcgaacaaca tcggtcatgc caacgggtct
     3121 cgtctctcga gcccttttgg cggcgactaa gaagtatgaa gcttcaggcc gcaacgcgcg
     3181 acacagcgtt ttgtgtggtg ggcctcggca ttgctctttg catggcccag cgtgattagt
     3241 gcgtggattt caagcccgag accgaaggat tgcgacatgt gcctggctgt ataactcacg
     3301 cttgctgcta cgctcgcctc ctcctccatc cactccatcg ccatggcctc tagtggatca
     3361 gcttgcatgc ctgcaggaga cgatatcacc tcttctgttt ccacgataaa aatagactgc
     3421 tcatttcttc gtcgtcttca tcgtctgctt tttctgcttc gcctctgtct ggggtctgaa
     3481 accactacac acacacacaa cactcgtact cccactttca caaaagcgta agctcaccgg
     3541 cttttcttac acgtacattt tagtggatcc catcacgcca ctaccacgcc cgcgggggat
     3601 ggaacggagg ggagagagag aagggggaag catggatgaa tgagacattg agggaaagga
     3661 ggggagggag cagtccatca gggcgctacc tctcttgtcc cccaaaccct gttgagccgt
     3721 tcaacatgtt tcatgtttcc tccctccccc cttccctccc tggcctttcc gcggagccat
     3781 tcaagtgacg tctggaccgc accgtaacaa aatcgtttct atggggggtt tgtttgacaa
     3841 ccacgtcttc agcgttttta aaaaaaaaag cgggcaagcc ctctcaccct cactcatgcc
     3901 catcctcctc ctctcctgcg gaacattctt acaaaaggcg taactcgacg acaactcaaa
     3961 gaacgacaaa catcaatccc aaaaaaaaaa tctctactcg tctctcttgg atctttccaa
     4021 ttgtcagacc ttcctcttct ttttcgggct agccaccatg gacgcgtagt cgggcacgtc
     4081 gtaggggtat ccggccagga tgcgctcgca caggcgccag ccggttaccc cgttgatggt
     4141 cacgcggaac aacagcgagc catccgggtt gatcagccgc tcgtcaatga tcttattgcc
     4201 gttccacagg gtgccggtca ccgtgatctt cttgccgtcg aacacggcaa tgccctcgta
     4261 aggacggccg aagtagtcaa tcatgttcgg cgtcacaccg tcgatgacca gcgtcccgta
     4321 gtgcagtatc accttgaagt ggtgatcgtc cacggggtac acaaccttga agatcttctc
     4381 gatctggccc atttggtccc cgctcaggcc ctcgtaggga atgatgacgt ggatgtcgat
     4441 cttcaggccg ttctcgccgc tcaggacgat gcgctggatt ggggtcacgg agacgcccag
     4501 gttctggaaa agcgaggaca caccgccctg ctccagcacc tggtcgaggt tatagccagc
     4561 tgtctgccgc cagtcgccca cgaagtcctc gagagtaaac accatgttaa cggacccgct
     4621 gccggacccg tcagccctgc tgtctccacc gagctgagag aggtcgattc ttgtttcata
     4681 gagccccgta attgactgat gaatcagtgt ggcgtccagg acctcctttg tagaggtgta
     4741 ccgctttctg tctatggtgg tgtcgaagta cttgaaggct gcaggcgcgc ccaagttggt
     4801 cagagtaaac aagtggataa tgttttctgc ctgctccctg atgggcttat ccctgtgctt
     4861 attgtaagca gaaagcacct tatcgaggtt agcgtcggcg aggatcactc ttttggagaa
     4921 ttcgcttatt tgctcgatga tctcatcaag gtagtgtttg tgttgttcca cgaacagctg
     4981 cttctgctca ttatcttcgg gagacccttt gagcttttca tagtggctgg ccagatacaa
     5041 gaaattaacg tatttagagg gcagtgccag ctcgttacct ttctgcagct cgcccgcact
     5101 agcgagcatt cgtttccggc cgttttcaag ctcaaagaga gagtacttgg gaagcttaat
     5161 gatgaggtct tttttgacct ctttatatcc tttcgcctcg agaaagtcga tggggttttt
     5221 ttcgaagctt gatcgctcca tgattgtgat gcccagcagt tccttgacgc ttttgagttt
     5281 tttagacttc cctttctcca ctttggccac aaccagtaca ctgtaagcga ctgtaggaga
     5341 atcgaatccg ccgtatttct tggggtccca atcttttttg cgtgcgatca gcttgtcgct
     5401 gttccttttc gggaggatac tttccttgga gaagcctccg gtctgtactt cggtcttttt
     5461 aacgatgttc acctgcggca tggacaggac cttccggact gtcgcgaaat ccctaccctt
     5521 gtcccacacg atttctcctg tttctccgtt tgtttcgata agtggtcgct tccgaatctc
     5581 tccattggcc agtgtaatct cggtcttgaa aaaattcata atattgctgt aaaagaagta
     5641 cttagcggtg gccttgccta tttcctgctc agactttgcg atcattttcc taacatcgta
     5701 cactttatag tctccgtaaa caaattcaga ttcaagcttg ggatattttt tgataagtgc
     5761 agtgcctacc actgcattca ggtaggcatc atgcgcatgg tggtaattgt tgatctctct
     5821 caccttataa aactgaaagt cctttctgaa atctgagacc agcttagact tcagagtaat
     5881 aactttcacc tctcgaatca gtttgtcatt ttcatcgtac ttggtgttca tgcgtgaatc
     5941 gagaatttgg gccacgtgct tggtgatctg gcgtgtctca acaagctgcc ttttgatgaa
     6001 gccggcttta tccaactcag acaggccacc tcgttcagcc ttagtcagat tatcgaactt
     6061 ccgttgtgtg atcagtttgg cgttcagcag ctgccgccaa taatttttca ttttcttgac
     6121 aacttcttct gaggggacgt tatcactctt ccctctattt ttatcggatc ttgtcaacac
     6181 tttattatca atagaatcat ctttgagaaa agactggggc acgatagcat ccacgtcgta
     6241 gtcggagagc cgattgatgt ccagttcctg atccacgtac atgtccctgc cgttctgcag
     6301 gtagtacagg tagagcttct cattctgaag ctgggtgttt tcaactgggt gttccttaag
     6361 gatttgggac cccagttctt ttataccctc ttcaatcctc ttcatccttt ccctactgtt
     6421 cttctgtccc ttctgggtag tttggttctc tcgggccatc tcgataacga tattctcggg
     6481 cttatgcctt cccattactt tgacgagttc atccacgacc ttaacggtct gcagtattcc
     6541 ctttttgata gctgggctac ctgcaagatt agcgatgtgc tcgtgaagac tgtccccctg
     6601 gccagaaact tgtgctttct ggatgtcctc cttaaaggtg agagagtcat catggatcaa
     6661 ctgcatgaag ttccggttgg caaatccatc ggacttaaga aaatccagga ttgtctttcc
     6721 actctgcttg tctcggatcc cattgatcag ttttcttgac agccgccccc atcctgtata
     6781 tcggcgcctc ttgagctgtt tcatgacttt gtcgtcgaag agatgagcgt aagttttcaa
     6841 gcgttcttca atcatctccc tatcttcaaa caacgtaagg gtgaggacaa tgtcctcaag
     6901 aatgtcctcg ttctcctcat tgtccaggaa gtccttgtct ttaatgattt tcaggagatc
     6961 gtgatacgtt cccagggatg cgttgaagcg atcctccact ccgctgattt caacagagtc
     7021 gaaacattca atctttttga aatagtcttc tttgagctgt ttcacggtaa ctttccggtt
     7081 cgtcttgaag aggaggtcca cgatagcttt cttctgctct ccagacagga atgctggctt
     7141 tctcatccct tctgtgacgt atttgacctt ggtgagctcg ttataaactg tgaagtactc
     7201 gtacagcaga gagtgtttag gaagcacctt ttcgttaggc agatttttat caaagttagt
     7261 catcctttcg atgaaggact gggcagaggc ccccttatcc acgacttcct cgaagttcca
     7321 gggagtgatg gtctcttctg atttgcgagt catccacgcg aatctggaat ttccccgggc
     7381 gagggggcct acatagtagg gtatccgaaa tgtgaggatt ttctcaatct tttccctgtt
     7441 atctttcaaa aaggggtaga aatcctcttg ccgcctgagg atagcgtgca gttcgcccag
     7501 gtgaatctgg tgggggatgc ttccattgtc gaaagtgcgc tgtttgcgca acagatcttc
     7561 tctgttaagc tttaccagca gctcctcggt gccgtccatt ttttccaaga tgggcttaat
     7621 aaatttgtaa aattcctcct ggcttgctcc gccgtcaatg tatccggcgt agccattttt
     7681 agactgatcg aagaaaattt ccttgtactt ctcaggcagt tgctgtctga caagggcctt
     7741 cagcaaagtc aagtcttggt ggtgctcatc atagcgcttg atcatactag cgctcagcgg
     7801 agctttggtg atctccgtgt tcactcgcag aatatcactc agcagaatgg cgtctgacag
     7861 gttctttgcc gccaaaaaaa ggtctgcgta ctggtcgccg atctgggcca gcagattgtc
     7921 gagatcatca tcgtaggtgt ctttgctcag ttgaagcttg gcatcttcgg ccaggtcgaa
     7981 gttagattta aagttggggg tcagcccgag tgacagggcg ataagattac caaacaggcc
     8041 gttcttcttc tccccaggga gctgtgcgat gaggttttcg agccgccggg atttggacag
     8101 cctagcgctc aggattgctt tggcgtcaac tccggatgcg ttgatcgggt tctcttcgaa
     8161 aagctgattg taagtctgaa ccagttggat aaagagtttg tcgacatcgc tgttgtctgg
     8221 gttcaggtcc ccctcgatga ggaagtgtcc ccgaaatttg atcatatgcg ccagcgcgag
     8281 atagatcaac cgcaagtcag ccttatcagt actgtctaca agcttcttcc tcagatgata
     8341 tatggttggg tacttttcat ggtacgccac ctcgtccacg atattgccaa agattgggtg
     8401 gcgctcgtgc tttttatcct cctccaccaa aaaggactcc tccagcctat ggaagaaaga
     8461 gtcatccacc ttagccatct cattactaaa gatctcctgc aggtagcaga tccgattctt
     8521 tctgcgggta tatctgcgcc gtgctgttct tttgagccgc gtggcttcgg ccgtctcccc
     8581 ggagtcgaac aggagggcgc caatgaggtt cttctttatg ctgtggcgat cggtattgcc
     8641 cagaactttg aattttttgc tcggcacctt gtactcgtcc gtaatgacgg cccagccgac
     8701 gctgtttgtg ccgatagcga gcccaatgga gtacttcttg tccaccttcc gctttttctt
     8761 gggcatgctc gagggttgcg tgtgtatctg tgtgcagtgg atattgttac cgagtttggt
     8821 gagcgtgagt ccgttagtgc cctggtggtg gtggattagg agagtgggtg actcggtgtc
     8881 catggctttc ttcgctcatt ataggagggg aaaggaatga gggagggtgg ggagaccgcg
     8941 tctgttgttg accaccgatt tacttcttgc ctcccttccc ctccctcccc tcaatccgta
     9001 cgacacaaat agtagccgag tgtctgctgc agagcgcatg attagtgtgg tagacaacga
     9061 gggagggaag gatgtacagg gcatggcacg gagaagcgat ggtggccagg aagaggagag
     9121 gtcgcgagaa caggatgtgt tgcgaatgga taaaaacaga aagcgatggc tctgggcttc
     9181 gaaagcaggg gacattagga cgtgtagacc atctcgacgg atccctctgt atctctgttg
     9241 tgcgtgaatg ttttctgtgc acgtgtagtg tgtgagagta gaacccggga actcgaacag
     9301 agaaaagcat gggtggctgt ggtgtggagg cttcgttccc accacatgcc cttctccttc
     9361 gcctcgcctc tccctgcctt cttccacgca cccttgcgcc cctcgttctc aatacctggc
     9421 tcacttccac cattcaaaca accatcacga tacaggcatt tatctatcgt tgaagacttc
     9481 ttcctccggt agatcttagc caaggtaaga agaggggcat gcagcaagga gaaagaaatg
     9541 atgcatgatg aggaatagaa ggggaggagg gagggatatg atgggaagcg aaagcgcata
     9601 ttctggtggt ctgctgcctg atggggacgc gtctagctgt gacactgagg acggtggctg
     9661 ctggtggctg cgggcgctgc cttggtgatc aatgggagta aagggaggga aggaggtagc
     9721 gtgaacggat gacgcggaga agtttagggg tctctttacg tatcgcccct gccgcccgcc
     9781 tctctgcgat aaatgtgcct gttaccctgc agcctctatt cttcactgtg ttcctgtttt
     9841 ccaacagcct ctattcttcc ctgtcttttg ttgcagtggc gtcatcctct ctttgcccca
     9901 gtcgtcgttc tctcgactca ctcactcccc ccctccttcc ctccctccat ccacagaatc
     9961 gatgaagagc caccccctga tgagtccgtg aggacgaaac gagtaagctc gtcggggtgc
    10021 ttcacgtacg cctgttttag agctagaaat agcaagttaa aataaggcta gtccgttatc
    10081 aacttgaaaa agtggcaccg agtcggtgct tttggccggc atggtcccag cctcctcgct
    10141 ggcgccggct gggcaacatg cttcggcatg gcgaatggga ctcctgggta ccatgggaaa
    10201 gaaaggatga gaaaggagaa aggacatcta gataaccggc atatcacggt ggtgtattag
    10261 tgtagaatag tgaagagaag acttgggaaa atgtgtagga aaggttgttt ctgtgtatgt
    10321 gggttgggat gggtggctgt ttgagaagga acagcgggca gggcgatgta gtgctgaacg
    10381 ggcacggaac cactgagact gaaggaagta gggagagaga ggggcagggg acgtgcactt
    10441 taatctttgc ctcggtagag tatacccatg caagagtatg tggccacctg tggtggcttt
    10501 tggccaggtc tggtgcagtg ccaatcatct cccatcaata atacaacttc agaacaacgg
    10561 cgcattgatg gggagagaga aagtaaattt aagtaagggg tacgtagtag aggattcaac
    10621 tgaaatattt tcgaggagcg gttgggaagt tgaccgattg aaaggagaag ggaggggagc
    10681 aggtgtgata gtcatgtgta aagtaattct tttttgccgt cgtcacacaa tccacatcaa
    10741 tgataaaata tgtttaagga tcaatcacac ggagtcggtc ataaggcaac cgcaaacgca
    10801 atgcaaacta gcaagcaagc aggaccacaa caacaaccat taccatcaca ggcgacagca
    10861 gcagcagctg cagcagcagc aaggcaagca aaggccatct gcgcgtggac ttttccaggc
    10921 accggcttta agcgtaattt tcaatgattg ttgtccgtgt atcctctcac cctttcaccg
    10981 cttgcgcgca cgtgagcagc acaactggta atgcccgagg acaatagacg aggaaaaaga
    11041 agaagaaaaa tgaagaggga gtgatgaaga agaagagaag aataaaagaa gacagaaaag
    11101 aagataaaaa agtcaaaaga cgacaaagac ggaaaaaaaa gaaacgcgca attttaacac
    11161 cagcaacgac gacgaaaagg gcgtcagctt tgcaccggtt tcttagacgg atcgcttgcc
    11221 tgtaacttac acgcgcctcg tatcttttaa tgatggaata atttgggaat ttactctgtg
    11281 tttatttatt tttatgtttt gtatttggat tttagaaagt aaataaagaa ggtagaagag
    11341 ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa atttcaacaa aaagcgtact
    11401 ttacatatat atttattaga caagaaaagc agattaaata gatatacatt cgattaacga
    11461 taagtaaaat gtaaaatcac aggattttcg tgtgtggtct tctacacaga caagatgaaa
    11521 caattcggca ttaatacctg agagcaggaa gtacaagata aaaggtagta tttgttggcg
    11581 atccccctag agtcttttac atcttcggaa aacaaaaact attttttctt taattt
//
