LOCUS       sequence-347050-       12023 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pNOC episomal plasmid harboring the dead version of humanized spCas9
            gene sequence tagged to the KRAB domain and 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 12023)
  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 12023)
  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-KRAB_sgRNA Td2
FEATURES             Location/Qualifiers
     source          1..12023
                     /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(4771..4965)
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEPWLV"
     CDS             complement(join(5029..6543,6544..6546,6547..6612,6613..6618,
                     6619..9102,9103..9105,9106..9129))
                     /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(5029..6612,6613..6615,6616..9102,9103..9105,
                     9106..9129))
                     /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(5029..9129)
                     /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(9130..9150)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(9371..9382)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     ncRNA           10364..10400
                     /product="self-cleaving hammerhead ribozyme (Ferre-D'Amare 
                     and Scott, 2010)"
                     /label=hammerhead ribozyme
                     /ncRNA_class="hammerhead_ribozyme"
     misc_RNA        10421..10496
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     ncRNA           10501..10568
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     misc_feature    join(11597..12023,1..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 tataacgaat 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 gccggacccc cacaagtcgt tacgggccat accctccatc ttgatatcgc aagactgctt
     4681 gtccttaaaa atgctacgag aagagactga agatttaatt tcaaaagctg tttcagaatc
     4741 agggtgggtt tcttgatgaa tttccctctc aactagccag ggttcttcgc ctttctctaa
     4801 tctcagtatg acatctggct tggtcagttg ataaccaagg gaaactaaat ttttgtagtt
     4861 ttccaacata acgttcctgt aaacaatttg ctgggcagtg tcaagtagtt tccactcctc
     4921 acgagtaaag tccacgaaaa catccttaaa tgtgaccaac gttctgctcc aggctgtcaa
     4981 agacttcgcg tcgttaacgg acccgctgcc ggacccgtca gccctgctgt ctccaccgag
     5041 ctgagagagg tcgattcttg tttcatagag ccccgtaatt gactgatgaa tcagtgtggc
     5101 gtccaggacc tcctttgtag aggtgtaccg ctttctgtct atggtggtgt cgaagtactt
     5161 gaaggctgca ggcgcgccca agttggtcag agtaaacaag tggataatgt tttctgcctg
     5221 ctccctgatg ggcttatccc tgtgcttatt gtaagcagaa agcaccttat cgaggttagc
     5281 gtcggcgagg atcactcttt tggagaattc gcttatttgc tcgatgatct catcaaggta
     5341 gtgtttgtgt tgttccacga acagctgctt ctgctcatta tcttcgggag accctttgag
     5401 cttttcatag tggctggcca gatacaagaa attaacgtat ttagagggca gtgccagctc
     5461 gttacctttc tgcagctcgc ccgcactagc gagcattcgt ttccggccgt tttcaagctc
     5521 aaagagagag tacttgggaa gcttaatgat gaggtctttt ttgacctctt tatatccttt
     5581 cgcctcgaga aagtcgatgg ggtttttttc gaagcttgat cgctccatga ttgtgatgcc
     5641 cagcagttcc ttgacgcttt tgagtttttt agacttccct ttctccactt tggccacaac
     5701 cagtacactg taagcgactg taggagaatc gaatccgccg tatttcttgg ggtcccaatc
     5761 ttttttgcgt gcgatcagct tgtcgctgtt ccttttcggg aggatacttt ccttggagaa
     5821 gcctccggtc tgtacttcgg tctttttaac gatgttcacc tgcggcatgg acaggacctt
     5881 ccggactgtc gcgaaatccc tacccttgtc ccacacgatt tctcctgttt ctccgtttgt
     5941 ttcgataagt ggtcgcttcc gaatctctcc attggccagt gtaatctcgg tcttgaaaaa
     6001 attcataata ttgctgtaaa agaagtactt agcggtggcc ttgcctattt cctgctcaga
     6061 ctttgcgatc attttcctaa catcgtacac tttatagtct ccgtaaacaa attcagattc
     6121 aagcttggga tattttttga taagtgcagt gcctaccact gcattcaggt aggcatcatg
     6181 cgcatggtgg taattgttga tctctctcac cttataaaac tgaaagtcct ttctgaaatc
     6241 tgagaccagc ttagacttca gagtaataac tttcacctct cgaatcagtt tgtcattttc
     6301 atcgtacttg gtgttcatgc gtgaatcgag aatttgggcc acgtgcttgg tgatctggcg
     6361 tgtctcaaca agctgccttt tgatgaagcc ggctttatcc aactcagaca ggccacctcg
     6421 ttcagcctta gtcagattat cgaacttccg ttgtgtgatc agtttggcgt tcagcagctg
     6481 ccgccaataa tttttcattt tcttgacaac ttcttctgag gggacgttat cactcttccc
     6541 tctattttta tcggatcttg tcaacacttt attatcaata gaatcatctt tgagaaaaga
     6601 ctggggcacg atagcatcca cgtcgtagtc ggagagccga ttgatgtcca gttcctgatc
     6661 cacgtacatg tccctgccgt tctgcaggta gtacaggtag agcttctcat tctgaagctg
     6721 ggtgttttca actgggtgtt ccttaaggat ttgggacccc agttctttta taccctcttc
     6781 aatcctcttc atcctttccc tactgttctt ctgtcccttc tgggtagttt ggttctctcg
     6841 ggccatctcg ataacgatat tctcgggctt atgccttccc attactttga cgagttcatc
     6901 cacgacctta acggtctgca gtattccctt tttgatagct gggctacctg caagattagc
     6961 gatgtgctcg tgaagactgt ccccctggcc agaaacttgt gctttctgga tgtcctcctt
     7021 aaaggtgaga gagtcatcat ggatcaactg catgaagttc cggttggcaa atccatcgga
     7081 cttaagaaaa tccaggattg tctttccact ctgcttgtct cggatcccat tgatcagttt
     7141 tcttgacagc cgcccccatc ctgtatatcg gcgcctcttg agctgtttca tgactttgtc
     7201 gtcgaagaga tgagcgtaag ttttcaagcg ttcttcaatc atctccctat cttcaaacaa
     7261 cgtaagggtg aggacaatgt cctcaagaat gtcctcgttc tcctcattgt ccaggaagtc
     7321 cttgtcttta atgattttca ggagatcgtg atacgttccc agggatgcgt tgaagcgatc
     7381 ctccactccg ctgatttcaa cagagtcgaa acattcaatc tttttgaaat agtcttcttt
     7441 gagctgtttc acggtaactt tccggttcgt cttgaagagg aggtccacga tagctttctt
     7501 ctgctctcca gacaggaatg ctggctttct catcccttct gtgacgtatt tgaccttggt
     7561 gagctcgtta taaactgtga agtactcgta cagcagagag tgtttaggaa gcaccttttc
     7621 gttaggcaga tttttatcaa agttagtcat cctttcgatg aaggactggg cagaggcccc
     7681 cttatccacg acttcctcga agttccaggg agtgatggtc tcttctgatt tgcgagtcat
     7741 ccacgcgaat ctggaatttc cccgggcgag ggggcctaca tagtagggta tccgaaatgt
     7801 gaggattttc tcaatctttt ccctgttatc tttcaaaaag gggtagaaat cctcttgccg
     7861 cctgaggata gcgtgcagtt cgcccaggtg aatctggtgg gggatgcttc cattgtcgaa
     7921 agtgcgctgt ttgcgcaaca gatcttctct gttaagcttt accagcagct cctcggtgcc
     7981 gtccattttt tccaagatgg gcttaataaa tttgtaaaat tcctcctggc ttgctccgcc
     8041 gtcaatgtat ccggcgtagc catttttaga ctgatcgaag aaaatttcct tgtacttctc
     8101 aggcagttgc tgtctgacaa gggccttcag caaagtcaag tcttggtggt gctcatcata
     8161 gcgcttgatc atactagcgc tcagcggagc tttggtgatc tccgtgttca ctcgcagaat
     8221 atcactcagc agaatggcgt ctgacaggtt ctttgccgcc aaaaaaaggt ctgcgtactg
     8281 gtcgccgatc tgggccagca gattgtcgag atcatcatcg taggtgtctt tgctcagttg
     8341 aagcttggca tcttcggcca ggtcgaagtt agatttaaag ttgggggtca gcccgagtga
     8401 cagggcgata agattaccaa acaggccgtt cttcttctcc ccagggagct gtgcgatgag
     8461 gttttcgagc cgccgggatt tggacagcct agcgctcagg attgctttgg cgtcaactcc
     8521 ggatgcgttg atcgggttct cttcgaaaag ctgattgtaa gtctgaacca gttggataaa
     8581 gagtttgtcg acatcgctgt tgtctgggtt caggtccccc tcgatgagga agtgtccccg
     8641 aaatttgatc atatgcgcca gcgcgagata gatcaaccgc aagtcagcct tatcagtact
     8701 gtctacaagc ttcttcctca gatgatatat ggttgggtac ttttcatggt acgccacctc
     8761 gtccacgata ttgccaaaga ttgggtggcg ctcgtgcttt ttatcctcct ccaccaaaaa
     8821 ggactcctcc agcctatgga agaaagagtc atccacctta gccatctcat tactaaagat
     8881 ctcctgcagg tagcagatcc gattctttct gcgggtatat ctgcgccgtg ctgttctttt
     8941 gagccgcgtg gcttcggccg tctccccgga gtcgaacagg agggcgccaa tgaggttctt
     9001 ctttatgctg tggcgatcgg tattgcccag aactttgaat tttttgctcg gcaccttgta
     9061 ctcgtccgta atgacggccc agccgacgct gtttgtgccg atagcgagcc caatggagta
     9121 cttcttgtcc accttccgct ttttcttggg catgctcgag ggttgcgtgt gtatctgtgt
     9181 gcagtggata ttgttaccga gtttggtgag cgtgagtccg ttagtgccct ggtggtggtg
     9241 gattaggaga gtgggtgact cggtgtccat ggctttcttc gctcattata ggaggggaaa
     9301 ggaatgaggg agggtgggga gaccgcgtct gttgttgacc accgatttac ttcttgcctc
     9361 ccttcccctc cctcccctca atccgtacga cacaaatagt agccgagtgt ctgctgcaga
     9421 gcgcatgatt agtgtggtag acaacgaggg agggaaggat gtacagggca tggcacggag
     9481 aagcgatggt ggccaggaag aggagaggtc gcgagaacag gatgtgttgc gaatggataa
     9541 aaacagaaag cgatggctct gggcttcgaa agcaggggac attaggacgt gtagaccatc
     9601 tcgacggatc cctctgtatc tctgttgtgc gtgaatgttt tctgtgcacg tgtagtgtgt
     9661 gagagtagaa cccgggaact cgaacagaga aaagcatggg tggctgtggt gtggaggctt
     9721 cgttcccacc acatgccatt ctccttcgcc tcgcctctcc ctgccttctt ccacgcaccc
     9781 ttgcgcccct cgttctcaat acctggctca cttccaccat tcaaacaacc atcacgatac
     9841 aggcatttat ctatcgttga agacttcttc ctccggtaga tcttagccaa ggtaagaaga
     9901 ggggcatgca gcaaggagaa agaaatgatg catgatgagg aatagaaggg gaggagggag
     9961 ggatatgatg ggaagcgaaa gcgcatattc tggtggtctg ctgcctgatg gggacgcgtc
    10021 tagctgtgac actgaggacg gtggctgctg gtggctgcgg gcgctgcctt ggtgatcaat
    10081 gggagtaaag ggagggaagg aggtagcgtg aacggatgac gcggagaagt ttaggggtct
    10141 ctttacgtat cgcccctgcc gcccgcctct ctgcgataaa tgtgcctgtt accctgcagc
    10201 ctctattctt cactgtgttc ctgttttcca acagcctcta ttcttccctg tcttttgttg
    10261 cagtggcgtc atcctctctt tgccccagtc gtcgttctct cgactcactc actccccccc
    10321 tccttccctc cctccatcca cagaatcgat gaagagccac cccctgatga gtccgtgagg
    10381 acgaaacgag taagctcgtc ggggtgcttc acgtacgcct gttttagagc tagaaatagc
    10441 aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt
    10501 ggccggcatg gtcccagcct cctcgctggc gccggctggg caacatgctt cggcatggcg
    10561 aatgggactc ctgggtacca tgggaaagaa aggatgagaa aggagaaagg acatctagat
    10621 aaccggcata tcacggtggt gtattagtgt agaatagtga agagaagact tgggaaaatg
    10681 tgtaggaaag gttgtttctg tgtatgtggg ttgggatggg tggctgtttg agaaggaaca
    10741 gcgggcaggg cgatgtagtg ctgaacgggc acggaaccac tgagactgaa ggaagtaggg
    10801 agagagaggg gcaggggacg tgcactttaa tctttgcctc ggtagagtat acccatgcaa
    10861 gagtatgtgg ccacctgtgg tggcttttgg ccaggtctgg tgcagtgcca atcatctccc
    10921 atcaataata caacttcaga acaacggcgc attgatgggg agagagaaag taaatttaag
    10981 taaggggtac gtagtagagg attcaactga aatattttcg aggagcggtt gggaagttga
    11041 ccgattgaaa ggagaaggga ggggagcagg tgtgatagtc atgtgtaaag taattctttt
    11101 ttgccgtcgt cacacaatcc acatcaatga taaaatatgt ttaaggatca atcacacgga
    11161 gtcggtcata aggcaaccgc aaacgcaatg caaactagca agcaagcagg accacaacaa
    11221 caaccattac catcacaggc gacagcagca gcagctgcag cagcagcaag gcaagcaaag
    11281 gccatctgcg cgtggacttt tccaggcacc ggctttaagc gtaattttca atgattgttg
    11341 tccgtgtatc ctctcaccct ttcaccgctt gcgcgcacgt gagcagcaca actggtaatg
    11401 cccgaggaca atagacgagg aaaaagaaga agaaaaatga agagggagtg atgaagaaga
    11461 agagaagaat aaaagaagac agaaaagaag ataaaaaagt caaaagacga caaagacgga
    11521 aaaaaaagaa acgcgcaatt ttaacaccag caacgacgac gaaaagggcg tcagctttgc
    11581 accggtttct tagacggatc gcttgcctgt aacttacacg cgcctcgtat cttttaatga
    11641 tggaataatt tgggaattta ctctgtgttt atttattttt atgttttgta tttggatttt
    11701 agaaagtaaa taaagaaggt agaagagtta cggaatgaag aaaaaaaaat aaacaaaggt
    11761 ttaaaaaatt tcaacaaaaa gcgtacttta catatatatt tattagacaa gaaaagcaga
    11821 ttaaatagat atacattcga ttaacgataa gtaaaatgta aaatcacagg attttcgtgt
    11881 gtggtcttct acacagacaa gatgaaacaa ttcggcatta atacctgaga gcaggaagta
    11941 caagataaaa ggtagtattt gttggcgatc cccctagagt cttttacatc ttcggaaaac
    12001 aaaaactatt ttttctttaa ttt
//
