LOCUS       sequence-228202-       11111 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level2 MoClo construct, containing level1 plant expression cassettes
            for SpCas-p(pEPOR1CB0002) and sgRNA_NbPDS(pEPOR1CB0114).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11111)
  AUTHORS   Raitskin O, Schudoma C, West A, Patron NJ
  TITLE     Comparison of efficiency and specificity of CRISPR-associated (Cas) 
            nucleases in plants: An expanded toolkit for precision genome 
            engineering.
  JOURNAL   PLoS One. 2019 Feb 27;14(2):e0211598. doi: 
            10.1371/journal.pone.0211598. eCollection 2019.
  PUBMED    30811422
REFERENCE   2  (bases 1 to 11111)
  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"; journalName: "PLoS One.";
            date: "2019-02-27"; volume: "14(2):e0211598. doi"; pages: " 
            10.1371/journal.pone.0211598. eCollection 2019"
COMMENT     Sequence Label: pEPOR2KN0002
FEATURES             Location/Qualifiers
     source          1..11111
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        454..799
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     742..762
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             890..4993
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             890..4993
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5006..5026
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(5033..5035,5036..5038,5039..5752)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5078..5099)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5339..5358)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5686..5707
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      5765..6017
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_RNA        6269..6344
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(6356..6380)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    6449..6473
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             6782..7411
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             7840..8913
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGASSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      8979..9173
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     promoter        9235..9339
                     /gene="bla"
                     /label=AmpR promoter
     CDS             9340..10134
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      10305..10893
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10794..10813
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    11029..11053
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 gaacactctg tgccgatgta tcacctctcg caagaattca agcttggagg tcaacatggt
       61 ggagcacgac actctggtct actccaaaaa tgtcaaagat acagtctcag aagatcaaag
      121 ggctattgag acttttcaac aaaggataat ttcgggaaac ctcctcggat tccattgccc
      181 agctatctgt cacttcatcg aaaggacagt agaaaaggaa ggtggctcct acaaatgcca
      241 tcattgcgat aaaggaaagg ctatcattca agatctctct gccgacagtg gtcccaaaga
      301 tggaccccca cccacgagga gcatcgtgga aaaagaagag gttccaacca cgtctacaaa
      361 gcaagtggat tgatgtgata acatggtgga gcacgacact ctggtctact ccaaaaatgt
      421 caaagataca gtctcagaag atcaaagggc tattgagact tttcaacaaa ggataatttc
      481 gggaaacctc ctcggattcc attgcccagc tatctgtcac ttcatcgaaa ggacagtaga
      541 aaaggaaggt ggctcctaca aatgccatca ttgcgataaa ggaaaggcta tcattcaaga
      601 tctctctgcc gacagtggtc ccaaagatgg acccccaccc acgaggagca tcgtggaaaa
      661 agaagaggtt ccaaccacgt ctacaaagca agtggattga tgtgacatct ccactgacgt
      721 aagggatgac gcacaatccc actatccttc gcaagaccct tcctctatat aaggaagttc
      781 atttcatttg gagaggacac gctcgagtat aagagctcat ttttacaaca attaccaaca
      841 acaacaaaca acaaacaaca ttacaattac atttacaatt atcgatacaa tggataagaa
      901 gtactctatc ggactcgata tcggaactaa ctctgtggga tgggctgtga tcaccgatga
      961 gtacaaggtg ccatctaaga agttcaaggt tctcggaaac accgataggc actctatcaa
     1021 gaaaaacctt atcggtgctc tcctcttcga ttctggtgaa actgctgagg ctaccagact
     1081 caagagaacc gctagaagaa ggtacaccag aagaaagaac aggatctgct acctccaaga
     1141 gatcttctct aacgagatgg ctaaagtgga tgattcattc ttccacaggc tcgaagagtc
     1201 attcctcgtg gaagaagata agaagcacga gaggcaccct atcttcggaa acatcgttga
     1261 tgaggtggca taccacgaga agtaccctac tatctaccac ctcagaaaga agctcgttga
     1321 ttctactgat aaggctgatc tcaggctcat ctacctcgct ctcgctcaca tgatcaagtt
     1381 cagaggacac ttcctcatcg agggtgatct caaccctgat aactctgatg tggataagtt
     1441 gttcatccag ctcgtgcaga cctacaacca gcttttcgaa gagaacccta tcaacgcttc
     1501 aggtgtggat gctaaggcta tcctctctgc taggctctct aagtcaagaa ggcttgagaa
     1561 cctcattgct cagctccctg gtgagaagaa gaacggactt ttcggaaact tgatcgctct
     1621 ctctctcgga ctcaccccta acttcaagtc taacttcgat ctcgctgagg atgcaaagct
     1681 ccagctctca aaggatacct acgatgatga tctcgataac ctcctcgctc agatcggaga
     1741 tcagtacgct gatttgttcc tcgctgctaa gaacctctct gatgctatcc tcctcagtga
     1801 tatcctcaga gtgaacaccg agatcaccaa ggctccactc tcagcttcta tgatcaagag
     1861 atacgatgag caccaccagg atctcacact tctcaaggct cttgttagac agcagctccc
     1921 agagaagtac aaagagattt tcttcgatca gtctaagaac ggatacgctg gttacatcga
     1981 tggtggtgca tctcaagaag agttctacaa gttcatcaag cctatcctcg agaagatgga
     2041 tggaaccgag gaactcctcg tgaagctcaa tagagaggat cttctcagaa agcagaggac
     2101 cttcgataac ggatctatcc ctcatcagat ccacctcgga gagttgcacg ctatccttag
     2161 aaggcaagag gatttctacc cattcctcaa ggataacagg gaaaagattg agaagattct
     2221 caccttcaga atcccttact acgtgggacc tctcgctaga ggaaactcaa gattcgcttg
     2281 gatgaccaga aagtctgagg aaaccatcac cccttggaac ttcgaagagg tggtggataa
     2341 gggtgctagt gctcagtctt tcatcgagag gatgaccaac ttcgataaga accttccaaa
     2401 cgagaaggtg ctccctaagc actctttgct ctacgagtac ttcaccgtgt acaacgagtt
     2461 gaccaaggtt aagtacgtga ccgagggaat gaggaagcct gcttttttgt caggtgagca
     2521 aaagaaggct atcgttgatc tcttgttcaa gaccaacaga aaggtgaccg tgaagcagct
     2581 caaagaggat tacttcaaga aaatcgagtg cttcgattca gttgagattt ctggtgttga
     2641 ggataggttc aacgcatctc tcggaaccta ccacgatctc ctcaagatca ttaaggataa
     2701 ggatttcttg gataacgagg aaaacgagga tatcttggag gatatcgttc ttaccctcac
     2761 cctctttgaa gatagagaga tgattgaaga aaggctcaag acctacgctc atctcttcga
     2821 tgataaggtg atgaagcagt tgaagagaag aagatacact ggttggggaa ggctctcaag
     2881 aaagctcatt aacggaatca gggataagca gtctggaaag acaatccttg atttcctcaa
     2941 gtctgatgga ttcgctaaca gaaacttcat gcagctcatc cacgatgatt ctctcacctt
     3001 taaagaggat atccagaagg ctcaggtttc aggacagggt gatagtctcc atgagcatat
     3061 cgctaacctc gctggatctc ctgcaatcaa gaagggaatc ctccagactg tgaaggttgt
     3121 ggatgagttg gtgaaggtga tgggaaggca taagcctgag aacatcgtga tcgaaatggc
     3181 tagagagaac cagaccactc agaagggaca gaagaactct agggaaagga tgaagaggat
     3241 cgaggaaggt atcaaagagc ttggatctca gatcctcaaa gagcaccctg ttgagaacac
     3301 tcagctccag aatgagaagc tctacctcta ctacctccag aacggaaggg atatgtatgt
     3361 ggatcaagag ttggatatca acaggctctc tgattacgat gttgatcata tcgtgccaca
     3421 gtcattcttg aaggatgatt ctatcgataa caaggtgctc accaggtctg ataagaacag
     3481 gggtaagagt gataacgtgc caagtgaaga ggttgtgaag aaaatgaaga actattggag
     3541 gcagctcctc aacgctaagc tcatcactca gagaaagttc gataacttga ctaaggctga
     3601 gaggggagga ctctctgaat tggataaggc aggattcatc aagaggcagc ttgtggaaac
     3661 caggcagatc actaagcacg ttgcacagat cctcgattct aggatgaaca ccaagtacga
     3721 tgagaacgat aagttgatca gggaagtgaa ggttatcacc ctcaagtcaa agctcgtgtc
     3781 tgatttcaga aaggatttcc aattctacaa ggtgagggaa atcaacaact accaccacgc
     3841 tcacgatgct taccttaacg ctgttgttgg aaccgctctc atcaagaagt atcctaagct
     3901 cgagtcagag ttcgtgtacg gtgattacaa ggtgtacgat gtgaggaaga tgatcgctaa
     3961 gtctgagcaa gagatcggaa aggctaccgc taagtatttc ttctactcta acatcatgaa
     4021 tttcttcaag accgagatta ccctcgctaa cggtgagatc agaaagaggc cactcatcga
     4081 gacaaacggt gaaacaggtg agatcgtgtg ggataaggga agggatttcg ctaccgttag
     4141 aaaggtgctc tctatgccac aggtgaacat cgttaagaaa accgaggtgc agaccggtgg
     4201 attctctaaa gagtctatcc tccctaagag gaactctgat aagctcattg ctaggaagaa
     4261 ggattgggac cctaagaaat acggtggttt cgattctcct accgtggctt actctgttct
     4321 cgttgtggct aaggttgaga agggaaagag taagaagctc aagtctgtta aggaacttct
     4381 cggaatcact atcatggaaa ggtcatcttt cgagaagaac ccaatcgatt tcctcgaggc
     4441 taagggatac aaagaggtta agaaggatct catcatcaag ctcccaaagt actcactctt
     4501 cgaactcgag aacggtagaa agaggatgct cgcttctgct ggtgagcttc aaaagggaaa
     4561 cgagcttgct ctcccatcta agtacgttaa ctttctttac ctcgcttctc actacgagaa
     4621 gttgaaggga tctccagaag ataacgagca gaagcaactt ttcgttgagc agcacaagca
     4681 ctacttggat gagatcatcg agcagatctc tgagttctct aaaagggtga tcctcgctga
     4741 tgcaaacctc gataaggtgt tgtctgctta caacaagcac agagataagc ctatcaggga
     4801 acaggcagag aacatcatcc atctcttcac ccttaccaac ctcggtgctc ctgctgcttt
     4861 caagtacttc gatacaacca tcgataggaa gagatacacc tctaccaaag aagtgctcga
     4921 tgctaccctc atccatcagt ctatcactgg actctacgag actaggatcg atctctcaca
     4981 gctcggtggt gattcaaggg ctgatcctaa gaagaagagg aaggttggtt cgatggtgag
     5041 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     5101 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     5161 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     5221 caccttcggc tacggcctgc agtgcttcgc ccgctacccc gaccacatga agcagcacga
     5281 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     5341 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     5401 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     5461 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     5521 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     5581 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     5641 ctaccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     5701 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt gagcttgtca
     5761 agcagatcgt tcaaacattt ggcaataaag tttcttaaga ttgaatcctg ttgccggtct
     5821 tgcgatgatt atcatataat ttctgttgaa ttacgttaag catgtaataa ttaacatgta
     5881 atgcatgacg ttatttatga gatgggtttt tatgattaga gtcccgcaat tatacattta
     5941 atacgcgata gaaaacaaaa tatagcgcgc aaactaggat aaattatcgc gcgcggtgtc
     6001 atctatgtta ctagatcgac gctactagaa ttcgagctcg gagcatcttc attcttaaga
     6061 tatgaagata atcttcaaaa ggcccctggg aatctgaaag aagagaagca ggcccattta
     6121 tatgggaaag aacaatagta tttcttatat aggcccattt aagttgaaaa caatcttcaa
     6181 aagtcccaca tcgcttagat aagaaaacga agctgagttt atatacagct agagtcgaag
     6241 tagtgattgc cgttaatttg agagtccagt tttagagcta gaaatagcaa gttaaaataa
     6301 ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt tctagaccca
     6361 gctttcttgt acaaagttgg cattacgctt tacgaggatg cacatgtgac cgagggacac
     6421 gaagtgatcc gtttaaacta tcagtgtttg acaggatata ttggcgggta aacctaagag
     6481 aaaagagcgt ttattagaat aatcggatat ttaaaagggc gtgaaaaggt ttatccgttc
     6541 gtccatttgt atgtgcatgc caaccacagg gttcccctcg ggagtcagcc gtgcggctgc
     6601 atgaaatcct ggccggtttg tctgatgcca agctggcggc ctggccggcc agcttggccg
     6661 ctgaagaaac cgagcgccgc cgtctaaaaa ggtgatgtgt atttgagtaa aacagcttgc
     6721 gtcatgcggt cgctgcgtat atgatgcgat gagtaaataa acaaatacgc aaggggaacg
     6781 catgaaggtt atcgctgtac ttaaccagaa aggcgggtca ggcaagacga ccatcgcaac
     6841 ccatctagcc cgcgccctgc aactcgccgg ggccgatgtt ctgttagtcg attccgatcc
     6901 ccagggcagt gcccgcgatt gggcggccgt gcgggaagat caaccgctaa ccgttgtcgg
     6961 catcgaccgc ccgacgattg accgcgacgt gaaggccatc ggccggcgcg acttcgtagt
     7021 gatcgacgga gcgccccagg cggcggactt ggctgtgtcc gcgatcaagg cagccgactt
     7081 cgtgctgatt ccggtgcagc caagccctta cgacatatgg gccaccgccg acctggtgga
     7141 gctggttaag cagcgcattg aggtcacgga tggaaggcta caagcggcct ttgtcgtgtc
     7201 gcgggcgatc aaaggcacgc gcatcggcgg tgaggttgcc gaggcgctgg ccgggtacga
     7261 gctgcccatt cttgagtccc gtatcacgca gcgcgtgagc tacccaggca ctgccgccgc
     7321 cggcacaacc gttcttgaat cagaacccga gggcgacgct gcccgcgagg tccaggcgct
     7381 ggccgctgaa attaaatcaa aactcatttg agttaatgag gtaaagagaa aatgagcaaa
     7441 agcacaaaca cgctaagtgc cggccgtccg agcgcacgca gcagcaaggc tgcaacgttg
     7501 gccagcctgg cagacacgcc agccatgaag cgggtcaact ttcagttgcc ggcggaggat
     7561 cacaccaagc tgaagatgta cgcggtacgc caaggcaaga ccattaccga gctgctatct
     7621 gaatacatcg cgcagctacc agagtaaatg agcaaatgaa taaatgagta gatgaatttt
     7681 agcggctaaa ggaggcggca tggaaaatca agaacaacca ggcaccgacg ccgtggaatg
     7741 ccccatgtgt ggaggaacgg gcggttggcc aggcgtaagc ggctgggttg tctgccggcc
     7801 ctgcaatggc actggaaccc ccaagcccga ggaatcggcg tgagcggtcg caaaccatcc
     7861 ggcccggtac aaatcggcgc ggcgctgggt gatgacctgg tggagaagtt gaaggccgcg
     7921 caggccgccc agcggcaacg catcgaggca gaagcacgcc ccggtgaatc gtggcaagcg
     7981 gccgctgatc gaatccgcaa agaatcccgg caaccgccgg cagccggtgc gtcgtcgatt
     8041 aggaagccgc ccaagggcga cgagcaacca gattttttcg ttccgatgct ctatgacgtg
     8101 ggcacccgcg atagtcgcag catcatggac gtggccgttt tccgtctgtc gaagcgtgac
     8161 cgacgagctg gcgaggtgat ccgctacgag cttccagacg ggcacgtaga ggtttccgca
     8221 gggccggccg gcatggccag tgtgtgggat tacgacctgg tactgatggc ggtttcccat
     8281 ctaaccgaat ccatgaaccg ataccgggaa gggaagggag acaagcccgg ccgcgtgttc
     8341 cgtccacacg ttgcggacgt actcaagttc tgccggcgag ccgatggcgg aaagcagaaa
     8401 gacgacctgg tagaaacctg cattcggtta aacaccacgc acgttgccat gcagcgtacg
     8461 aagaaggcca agaacggccg cctggtgacg gtatccgagg gtgaagcctt gattagccgc
     8521 tacaagatcg taaagagcga aaccgggcgg ccggagtaca tcgagatcga gctagctgat
     8581 tggatgtacc gcgagattac agaaggcaag aacccggacg tgctgacggt tcaccccgat
     8641 tactttttga tcgatcccgg catcggccgt tttctctacc gcctggcacg ccgcgccgca
     8701 ggcaaggcag aagccagatg gttgttcaag acgatctacg aacgcagtgg cagcgccgga
     8761 gagttcaaga agttctgttt caccgtgcgc aagctgatcg ggtcaaatga cctgccggag
     8821 tacgatttga aggaggaggc ggggcaggct ggcccgatcc tagtcatgcg ctaccgcaac
     8881 ctgatcgagg gcgaagcatc cgccggttcc taatgtacgg agcagatgct agggcaaatt
     8941 gccctagcag gggaaaaagg tcgaaaaagc ttctttcctg tggatagcac gtacattggg
     9001 aacccaaagc cgtacattgg gaaccggaac ccgtacattg ggaacccaaa gccgtacatt
     9061 gggaaccggt cacacatgta agtgactgat ataaaagaga aaaaaggcga tttttccgcc
     9121 taaaactctt taaaacttat taaaactctt aaaacccgcc tggcctgtgc ataactgtct
     9181 ggccagcgca cagccgaaca gctgcaaaaa gcgcccacat caaggctccg agtgcgcgga
     9241 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     9301 ccctgataaa tgcttcaata atattgaaaa aggaagagta tggctaaaat gagaatatca
     9361 ccggaattga aaaaactgat cgaaaaatac cgctgcgtaa aagatacgga aggaatgtct
     9421 cctgctaagg tatataagct ggtgggagaa aatgaaaacc tatatttaaa aatgacggac
     9481 agccggtata aagggaccac ctatgatgtg gaacgggaaa aggacatgat gctatggctg
     9541 gaaggaaagc tgcctgttcc aaaggtcctg cactttgaac ggcatgatgg ctggagcaat
     9601 ctgctcatga gtgaggccga tggcgtcctt tgctcggaag agtatgaaga tgaacaaagc
     9661 cctgaaaaga ttatcgagct gtatgcggag tgcatcaggc tctttcactc catcgacata
     9721 tcggattgtc cctatacgaa tagcttagac agccgcttag ccgaattgga ttacttactg
     9781 aataacgatc tggccgatgt ggattgcgaa aactgggaag aggacactcc atttaaagat
     9841 ccgcgcgagc tgtatgattt tttaaagacg gaaaagcccg aagaggaact tgtcttttcc
     9901 cacggcgacc tgggagacag caacatcttt gtgaaagatg gcaaagtaag tggctttatt
     9961 gatcttggga gaagcggcag ggcggacaag tggtatgaca ttgccttctg cgtccggtcg
    10021 ctcagggagg atatcgggga agaacagtat gtcgagctat tttttgactt actggggatc
    10081 aagcctgatt gggagaaaat aaaatattat attttactgg atgaattgtt ttagctgtca
    10141 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
    10201 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
    10261 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
    10321 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
    10381 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
    10441 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
    10501 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
    10561 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
    10621 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
    10681 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
    10741 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
    10801 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
    10861 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
    10921 ttcctgctcg gatctgttgg accggacagt agtcatggtt gatgggctgc ctgtatcgag
    10981 tggtgatttt gtgccgagct gccggtcggg gagctgttgg ctggctggtg gcaggatata
    11041 ttgtggtgta aacaaattga cgcttagaca acttaataac acattgcgga cgtttttaat
    11101 gtactggggt t
//
