LOCUS       sequence-346581-       11644 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pNOC episomal plasmid harboring the dead version of humanized spCas9
            gene sequence tagged with Nlux and spacer sequence on sgRNA is 
            replaced by the type IIS restriction site for endonuclease BaeI and.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11644)
  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 11644)
  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 NC
FEATURES             Location/Qualifiers
     source          1..11644
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             4..30
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      32..41
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             46..66
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     rep_origin      complement(2255..2843)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2335..2354)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3014..3805,3806..3874))
                     /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     3637..3656
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3875..3979)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    4016..4519
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     ncRNA           complement(5548..5615)
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     misc_RNA        complement(5620..5695)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     ncRNA           complement(5724..5760)
                     /product="self-cleaving hammerhead ribozyme (Ferre-D'Amare 
                     and Scott, 2010)"
                     /label=hammerhead ribozyme
                     /ncRNA_class="hammerhead_ribozyme"
     CDS             6742..6753
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             6974..6994
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(6995..7018,7019..7021,7022..9505,9506..9511,9512..9577,
                     9578..9580,9581..11095)
                     /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             join(6995..7018,7019..7021,7022..9508,9509..9511,
                     9512..11095)
                     /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             6995..11095
                     /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"
ORIGIN
        1 ggatacccct acgacgtgcc cgactacgcg tccatggtgg ctagcccgaa aaagaagagg
       61 aaggtctgac aattggaaag atccaagaga gacgagtaga gatttttttt ttgggattga
      121 tgtttgtcgt tctttgagtt gtcgtcgagt tacgcctttt gtaagaatgt tccgcaggag
      181 aggaggagga tgggcatgag tgagggtgag agggcttgcc cgcttttttt tttaaaaacg
      241 ctgaagacgt ggttgtcaaa caaacccccc atagaaacga ttttgttacg gtgcggtcca
      301 gacgtcactt gaatggctcc gcggaaaggc cagggaggga aggggggagg gaggaaacat
      361 gaaacatgtt gaacggctca acagggtttg ggggacaaga gaggtagcgc cctgatggac
      421 tgctccctcc cctcctttcc ctcaatgtct cattcatcca tgcttccccc ttctctctct
      481 cccctccgtt ccatcccccg cgggcgtggt agtggcgtga tgggatccac taaaatgtac
      541 gtgtaagaaa agccggtgag cttacgcttt tgtgaaagtg ggagtacgag tgttgtgtgt
      601 gtgtgtagtg gtttcagacc ccagacagag gcgaagcaga aaaagcagac gatgaagacg
      661 acgaagaaat gagcagtcta tttttatcgt ggaaacagaa gaggtgatat cgtctcctgc
      721 aggcatgcaa gctgatccac tagaggccat ggcgatggag tggatggagg aggaggcgag
      781 cgtagcagca agcgtgagtt atacagccag gcacatgtcg caatccttcg gtctcgggct
      841 tgaaatccac gcactaatca cgctgggcca tgcaaagagc aatgccgagg cccaccacac
      901 aaaacgctgt gtcgcgcgtt gcggcctgaa gcttcatact tcttagtcgc cgccaaaagg
      961 gctcgagaga cgagacccgt tggcatgacc gatgttgttc gacgcggttt gcttcgtcac
     1021 agtcgacgtg attcaggaat ctggagcctg cagatcattt ttttcagcct gatatcgttc
     1081 ttttccactg agaaccatca gaccaccttt tcttccattg tgtgaaggag taggagttgc
     1141 cgtgctgctt tgtgggagac atctgcgatg gtgaccagcc tcccgtcgtc tggtcgacgt
     1201 gacgagcctc ttcactgttc ttcgacggag agacgcaagc gagacggctc tagacctttt
     1261 ggacacgcat tctgtgtgtg aactagtgga cagtgatacc acgtctgaaa gctcaccact
     1321 gcccatggtg cagctacttg tcacaaagtt ttgactccgt cggtatcacc attcgcgctc
     1381 gtgtgcctgg ttgttccgcc acgccggcct gccccggggc ggggcaatat tctaaaatct
     1441 cacgcaaaac accgcactta cccctcacac atattcgtga tagaccacca ccaatctcag
     1501 cccgcatcaa cacaggaggg cccatgcccc tctcgcagga ggagagcacg ctgatcgaga
     1561 gggctacggc gacgattaac tcgattccta tctccgagga ttactcggtg gcttccgctg
     1621 ctctgtcgtc cgacggacgg atcttcacgg gggtcaacgt gtaccatttt acgggggggc
     1681 cttgcgcgga gctggtggtc ctggggaccg ccgctgctgc cgcggctggg aacctgacgt
     1741 gcattgtcgc catcgggaac gagaaccgcg ggattctgag cccctgcggg cggtgccggc
     1801 aggtcctgct cgacctccac cctgggatca aggcgattgt gaaggattcc gacgggcagc
     1861 ctacggcggt cggtatcaga gaacttctcc cttcgggcta cgtgtgggag agcgcttgaa
     1921 ctagtagtag atgccgaccg gagtccgcaa aaatcaccag tctctctcta caaatctatc
     1981 tctctctatt tttctccaga ataatgtgtg agtagttccc agataaggga attagggttc
     2041 ttatagggtt tcgctcatgt gttgagcata taagaaaccc ttagtatgta tttgtatttg
     2101 taaaatactt ctatcaataa aatttctaat tcctaaaacc aaaatccagt gacctggttt
     2161 aaaccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     2221 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2281 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2341 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2401 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     2461 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     2521 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     2581 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     2641 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     2701 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     2761 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     2821 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     2881 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     2941 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     3001 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3061 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     3121 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     3181 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     3241 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     3301 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     3361 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     3421 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     3481 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     3541 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     3601 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     3661 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     3721 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     3781 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     3841 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     3901 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     3961 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgggtcc ttttcatcac
     4021 gtgctataaa aataattata atttaaattt tttaatataa atatataaat taaaaataga
     4081 aagtaaaaaa agaaattaaa gaaaaaatag tttttgtttt ccgaagatgt aaaagactct
     4141 agggggatcg ccaacaaata ctacctttta tcttgtactt cctgctctca ggtattaatg
     4201 ccgaattgtt tcatcttgtc tgtgtagaag accacacacg aaaatcctgt gattttacat
     4261 tttacttatc gttaatcgaa tgtatatcta tttaatctgc ttttcttgtc taataaatat
     4321 atatgtaaag tacgcttttt gttgaaattt tttaaacctt tgtttatttt tttttcttca
     4381 ttccgtaact cttctacctt ctttatttac tttctaaaat ccaaatacaa aacataaaaa
     4441 taaataaaca cagagtaaat tcccaaatta ttccatcatt aaaagatacg aggcgcgtgt
     4501 aagttacagg caagcgatcc gtctaagaaa ccggtgcaaa gctgacgccc ttttcgtcgt
     4561 cgttgctggt gttaaaattg cgcgtttctt ttttttccgt ctttgtcgtc ttttgacttt
     4621 tttatcttct tttctgtctt cttttattct tctcttcttc ttcatcactc cctcttcatt
     4681 tttcttcttc tttttcctcg tctattgtcc tcgggcatta ccagttgtgc tgctcacgtg
     4741 cgcgcaagcg gtgaaagggt gagaggatac acggacaaca atcattgaaa attacgctta
     4801 aagccggtgc ctggaaaagt ccacgcgcag atggcctttg cttgccttgc tgctgctgca
     4861 gctgctgctg ctgtcgcctg tgatggtaat ggttgttgtt gtggtcctgc ttgcttgcta
     4921 gtttgcattg cgtttgcggt tgccttatga ccgactccgt gtgattgatc cttaaacata
     4981 ttttatcatt gatgtggatt gtgtgacgac ggcaaaaaag aattacttta cacatgacta
     5041 tcacacctgc tcccctccct tctcctttca atcggtcaac ttcccaaccg ctcctcgaaa
     5101 atatttcagt tgaatcctct actacgtacc ccttacttaa atttactttc tctctcccca
     5161 tcaatgcgcc gttgttctga agttgtatta ttgatgggag atgattggca ctgcaccaga
     5221 cctggccaaa agccaccaca ggtggccaca tactcttgca tgggtatact ctaccgaggc
     5281 aaagattaaa gtgcacgtcc cctgcccctc tctctcccta cttccttcag tctcagtggt
     5341 tccgtgcccg ttcagcacta catcgccctg cccgctgttc cttctcaaac agccacccat
     5401 cccaacccac atacacagaa acaacctttc ctacacattt tcccaagtct tctcttcact
     5461 attctacact aatacaccac cgtgatatgc cggttatcta gatgtccttt ctcctttctc
     5521 atcctttctt tcccatggta cccaggagtc ccattcgcca tgccgaagca tgttgcccag
     5581 ccggcgccag cgaggaggct gggaccatgc cggccaaaag caccgactcg gtgccacttt
     5641 ttcaagttga taacggacta gccttatttt aacttgctat ttctagctct aaaacccagc
     5701 tggatactag cgtcatggga tccgacgagc ttactcgttt cgtcctcacg gactcatcag
     5761 ggatccgctc ttcatcgatt ctgtggatgg agggagggaa ggaggggggg agtgagtgag
     5821 tcgagagaac gacgactggg gcaaagagag gatgacgcca ctgcaacaaa agacagggaa
     5881 gaatagaggc tgttggaaaa caggaacaca gtgaagaata gaggctgcag ggtaacaggc
     5941 acatttatcg cagagaggcg ggcggcaggg gcgatacgta aagagacccc taaacttctc
     6001 cgcgtcatcc gttcacgcta cctccttccc tccctttact cccattgatc accaaggcag
     6061 cgcccgcagc caccagcagc caccgtcctc agtgtcacag ctagacgcgt ccccatcagg
     6121 cagcagacca ccagaatatg cgctttcgct tcccatcata tccctccctc ctccccttct
     6181 attcctcatc atgcatcatt tctttctcct tgctgcatgc ccctcttctt accttggcta
     6241 agatctaccg gaggaagaag tcttcaacga tagataaatg cctgtatcgt gatggttgtt
     6301 tgaatggtgg aagtgagcca ggtattgaga acgaggggcg caagggtgcg tggaagaagg
     6361 cagggagagg cgaggcgaag gagaagggca tgtggtggga acgaagcctc cacaccacag
     6421 ccacccatgc ttttctctgt tcgagttccc gggttctact ctcacacact acacgtgcac
     6481 agaaaacatt cacgcacaac agagatacag agggatccgt cgagatggtc tacacgtcct
     6541 aatgtcccct gctttcgaag cccagagcca tcgctttctg tttttatcca ttcgcaacac
     6601 atcctgttct cgcgacctct cctcttcctg gccaccatcg cttctccgtg ccatgccctg
     6661 tacatccttc cctccctcgt tgtctaccac actaatcatg cgctctgcag cagacactcg
     6721 gctactattt gtgtcgtacg gattgagggg agggagggga agggaggcaa gaagtaaatc
     6781 ggtggtcaac aacagacgcg gtctccccac cctccctcat tcctttcccc tcctataatg
     6841 agcgaagaaa gccatggaca ccgagtcacc cactctccta atccaccacc accagggcac
     6901 taacggactc acgctcacca aactcggtaa caatatccac tgcacacaga tacacacgca
     6961 accctcgagc atgcccaaga aaaagcggaa ggtggacaag aagtactcca ttgggctcgc
     7021 tatcggcaca aacagcgtcg gctgggccgt cattacggac gagtacaagg tgccgagcaa
     7081 aaaattcaaa gttctgggca ataccgatcg ccacagcata aagaagaacc tcattggcgc
     7141 cctcctgttc gactccgggg agacggccga agccacgcgg ctcaaaagaa cagcacggcg
     7201 cagatatacc cgcagaaaga atcggatctg ctacctgcag gagatcttta gtaatgagat
     7261 ggctaaggtg gatgactctt tcttccatag gctggaggag tcctttttgg tggaggagga
     7321 taaaaagcac gagcgccacc caatctttgg caatatcgtg gacgaggtgg cgtaccatga
     7381 aaagtaccca accatatatc atctgaggaa gaagcttgta gacagtactg ataaggctga
     7441 cttgcggttg atctatctcg cgctggcgca tatgatcaaa tttcggggac acttcctcat
     7501 cgagggggac ctgaacccag acaacagcga tgtcgacaaa ctctttatcc aactggttca
     7561 gacttacaat cagcttttcg aagagaaccc gatcaacgca tccggagttg acgccaaagc
     7621 aatcctgagc gctaggctgt ccaaatcccg gcggctcgaa aacctcatcg cacagctccc
     7681 tggggagaag aagaacggcc tgtttggtaa tcttatcgcc ctgtcactcg ggctgacccc
     7741 caactttaaa tctaacttcg acctggccga agatgccaag cttcaactga gcaaagacac
     7801 ctacgatgat gatctcgaca atctgctggc ccagatcggc gaccagtacg cagacctttt
     7861 tttggcggca aagaacctgt cagacgccat tctgctgagt gatattctgc gagtgaacac
     7921 ggagatcacc aaagctccgc tgagcgctag tatgatcaag cgctatgatg agcaccacca
     7981 agacttgact ttgctgaagg cccttgtcag acagcaactg cctgagaagt acaaggaaat
     8041 tttcttcgat cagtctaaaa atggctacgc cggatacatt gacggcggag caagccagga
     8101 ggaattttac aaatttatta agcccatctt ggaaaaaatg gacggcaccg aggagctgct
     8161 ggtaaagctt aacagagaag atctgttgcg caaacagcgc actttcgaca atggaagcat
     8221 cccccaccag attcacctgg gcgaactgca cgctatcctc aggcggcaag aggatttcta
     8281 cccctttttg aaagataaca gggaaaagat tgagaaaatc ctcacatttc ggatacccta
     8341 ctatgtaggc cccctcgccc ggggaaattc cagattcgcg tggatgactc gcaaatcaga
     8401 agagaccatc actccctgga acttcgagga agtcgtggat aagggggcct ctgcccagtc
     8461 cttcatcgaa aggatgacta actttgataa aaatctgcct aacgaaaagg tgcttcctaa
     8521 acactctctg ctgtacgagt acttcacagt ttataacgag ctcaccaagg tcaaatacgt
     8581 cacagaaggg atgagaaagc cagcattcct gtctggagag cagaagaaag ctatcgtgga
     8641 cctcctcttc aagacgaacc ggaaagttac cgtgaaacag ctcaaagaag actatttcaa
     8701 aaagattgaa tgtttcgact ctgttgaaat cagcggagtg gaggatcgct tcaacgcatc
     8761 cctgggaacg tatcacgatc tcctgaaaat cattaaagac aaggacttcc tggacaatga
     8821 ggagaacgag gacattcttg aggacattgt cctcaccctt acgttgtttg aagataggga
     8881 gatgattgaa gaacgcttga aaacttacgc tcatctcttc gacgacaaag tcatgaaaca
     8941 gctcaagagg cgccgatata caggatgggg gcggctgtca agaaaactga tcaatgggat
     9001 ccgagacaag cagagtggaa agacaatcct ggattttctt aagtccgatg gatttgccaa
     9061 ccggaacttc atgcagttga tccatgatga ctctctcacc tttaaggagg acatccagaa
     9121 agcacaagtt tctggccagg gggacagtct tcacgagcac atcgctaatc ttgcaggtag
     9181 cccagctatc aaaaagggaa tactgcagac cgttaaggtc gtggatgaac tcgtcaaagt
     9241 aatgggaagg cataagcccg agaatatcgt tatcgagatg gcccgagaga accaaactac
     9301 ccagaaggga cagaagaaca gtagggaaag gatgaagagg attgaagagg gtataaaaga
     9361 actggggtcc caaatcctta aggaacaccc agttgaaaac acccagcttc agaatgagaa
     9421 gctctacctg tactacctgc agaacggcag ggacatgtac gtggatcagg aactggacat
     9481 caatcggctc tccgactacg acgtggatgc tatcgtgccc cagtcttttc tcaaagatga
     9541 ttctattgat aataaagtgt tgacaagatc cgataaaaat agagggaaga gtgataacgt
     9601 cccctcagaa gaagttgtca agaaaatgaa aaattattgg cggcagctgc tgaacgccaa
     9661 actgatcaca caacggaagt tcgataatct gactaaggct gaacgaggtg gcctgtctga
     9721 gttggataaa gccggcttca tcaaaaggca gcttgttgag acacgccaga tcaccaagca
     9781 cgtggcccaa attctcgatt cacgcatgaa caccaagtac gatgaaaatg acaaactgat
     9841 tcgagaggtg aaagttatta ctctgaagtc taagctggtc tcagatttca gaaaggactt
     9901 tcagttttat aaggtgagag agatcaacaa ttaccaccat gcgcatgatg cctacctgaa
     9961 tgcagtggta ggcactgcac ttatcaaaaa atatcccaag cttgaatctg aatttgttta
    10021 cggagactat aaagtgtacg atgttaggaa aatgatcgca aagtctgagc aggaaatagg
    10081 caaggccacc gctaagtact tcttttacag caatattatg aattttttca agaccgagat
    10141 tacactggcc aatggagaga ttcggaagcg accacttatc gaaacaaacg gagaaacagg
    10201 agaaatcgtg tgggacaagg gtagggattt cgcgacagtc cggaaggtcc tgtccatgcc
    10261 gcaggtgaac atcgttaaaa agaccgaagt acagaccgga ggcttctcca aggaaagtat
    10321 cctcccgaaa aggaacagcg acaagctgat cgcacgcaaa aaagattggg accccaagaa
    10381 atacggcgga ttcgattctc ctacagtcgc ttacagtgta ctggttgtgg ccaaagtgga
    10441 gaaagggaag tctaaaaaac tcaaaagcgt caaggaactg ctgggcatca caatcatgga
    10501 gcgatcaagc ttcgaaaaaa accccatcga ctttctcgag gcgaaaggat ataaagaggt
    10561 caaaaaagac ctcatcatta agcttcccaa gtactctctc tttgagcttg aaaacggccg
    10621 gaaacgaatg ctcgctagtg cgggcgagct gcagaaaggt aacgagctgg cactgccctc
    10681 taaatacgtt aatttcttgt atctggccag ccactatgaa aagctcaaag ggtctcccga
    10741 agataatgag cagaagcagc tgttcgtgga acaacacaaa cactaccttg atgagatcat
    10801 cgagcaaata agcgaattct ccaaaagagt gatcctcgcc gacgctaacc tcgataaggt
    10861 gctttctgct tacaataagc acagggataa gcccatcagg gagcaggcag aaaacattat
    10921 ccacttgttt actctgacca acttgggcgc gcctgcagcc ttcaagtact tcgacaccac
    10981 catagacaga aagcggtaca cctctacaaa ggaggtcctg gacgccacac tgattcatca
    11041 gtcaattacg gggctctatg aaacaagaat cgacctctct cagctcggtg gagacagcag
    11101 ggctgacggg tccggcagcg ggtccgttaa catggtgttt actctcgagg acttcgtggg
    11161 cgactggcgg cagacagctg gctataacct cgaccaggtg ctggagcagg gcggtgtgtc
    11221 ctcgcttttc cagaacctgg gcgtctccgt gaccccaatc cagcgcatcg tcctgagcgg
    11281 cgagaacggc ctgaagatcg acatccacgt catcattccc tacgagggcc tgagcgggga
    11341 ccaaatgggc cagatcgaga agatcttcaa ggttgtgtac cccgtggacg atcaccactt
    11401 caaggtgata ctgcactacg ggacgctggt catcgacggt gtgacgccga acatgattga
    11461 ctacttcggc cgtccttacg agggcattgc cgtgttcgac ggcaagaaga tcacggtgac
    11521 cggcaccctg tggaacggca ataagatcat tgacgagcgg ctgatcaacc cggatggctc
    11581 gctgttgttc cgcgtgacca tcaacggggt aaccggctgg cgcctgtgcg agcgcatcct
    11641 ggcc
//
