LOCUS       sequence-356747-       14182 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express SpCas9 and Kanamycin resistance gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14182)
  AUTHORS   Oh Y, Lee B, Kim H, Kim SG
  TITLE     A multiplex guide RNA expression system and its efficacy for plant 
            genome engineering.
  JOURNAL   Plant Methods. 2020 Mar 12;16:37. doi: 10.1186/s13007-020-00580-x. 
            eCollection 2020.
  PUBMED    32190101
REFERENCE   2  (bases 1 to 14182)
  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: "Plant 
            Methods."; date: "2020-03-12"; volume: "16:37. doi"; pages: " 
            10.1186/s13007-020-00580-x. eCollection 2020"
COMMENT     Sequence Label: pECO300
FEATURES             Location/Qualifiers
     source          1..14182
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..345
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     289..309
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     protein_bind    445..469
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             472..4575
                     /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             472..4575
                     /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             4591..4611
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4612..4638
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             4612..4638
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(4643..4667)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    4987..5011
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             6342..6971
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             7400..8473
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      8539..8733
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(8969..8991)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    9077..9217
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     9132..9151
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(9232..9249)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9403..9991)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9483..9502)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(10237..11028)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     misc_feature    11549..11573
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(11666..11918)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(12078..12872)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(12189..12208)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     12799..12818
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(12908..13087)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_RNA        complement(13191..13266)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 tgagactttt caacaaaggg taatatcggg aaacctcctc ggattccatt gcccagctat
       61 ctgtcacttc atcaaaagga cagtagaaaa ggaaggtggc acctacaaat gccatcattg
      121 cgataaagga aaggctatcg ttcaagatgc ccctgccgac agtggtccca aagatggacc
      181 cccacccacg aggagcatcg tggaaaaaga agacgttcca accacgtctt caaagcaagt
      241 ggattgatgt gatatctcca ctgacgtaag ggatgacgca caatcccact atccttcgca
      301 agacccttcc tctatataag gaagttcatt tcatttggag aggactccgg tatttttaca
      361 acaattacca caacaaaaca aacaacaaac aacattacaa tttactattc tagtcgacct
      421 gcaggcggcc gcactagtga tatcacaagt ttgtacaaaa aagcaggctt catggacaag
      481 aagtacagca tcggcctgga catcggtacc aacagcgtgg gctgggccgt gatcaccgac
      541 gagtacaagg tgcccagcaa gaagttcaag gtgctgggca acaccgaccg ccacagcatc
      601 aagaagaacc tgatcggcgc cctgctgttc gacagcggcg agaccgccga ggccacccgc
      661 ctgaagcgca ccgcccgccg ccgctacacc cgccgcaaga accgcatctg ctacctgcag
      721 gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccaccg cctggaggag
      781 agcttcctgg tggaggagga caagaagcac gagcgccacc ccatcttcgg caacatcgtg
      841 gacgaggtgg cctaccacga gaagtacccc accatctacc atctgcgcaa gaagctggtg
      901 gacagcaccg acaaggccga cctgcgcctg atctacctgg ccctggccca catgatcaag
      961 ttccgcggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag
     1021 ctgttcatcc agctggtgca gacctacaac cagctgttcg aggagaaccc catcaacgcc
     1081 agcggcgtgg acgccaaggc catcctgagc gcccgcctga gcaagagccg ccgcctggag
     1141 aacctgatcg cccagctgcc cggcgagaag aagaacggcc tgttcggcaa cctgatcgcc
     1201 ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga ggacgccaag
     1261 ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc
     1321 gaccagtacg ccgacctgtt cctggccgcc aagaacctga gcgacgccat cctgctgagc
     1381 gacatcctgc gcgtgaacac cgagatcacc aaggcccccc tgagcgccag catgatcaag
     1441 cgctacgacg agcaccacca ggacctgacc ctgctgaagg ccctggtgcg ccagcagctg
     1501 cccgagaagt acaaggagat cttcttcgac cagagcaaga acggctacgc cggctacatc
     1561 gacggcggcg ccagccagga ggagttctac aagttcatca agcccatcct ggagaagatg
     1621 gacggcaccg aggagctgct ggtgaagctg aaccgcgagg acctgctgcg caagcagcgc
     1681 accttcgaca acggcagcat cccccaccag atccacctgg gcgagctgca cgccatcctg
     1741 cgccgccagg aggacttcta ccccttcctg aaggacaacc gcgagaagat cgagaagatc
     1801 ctgaccttcc gcatccccta ctacgtgggc cccctggccc gcggcaacag ccgcttcgcc
     1861 tggatgaccc gcaagagcga ggagaccatc accccctgga acttcgagga ggtggtggac
     1921 aagggcgcca gcgcccagag cttcatcgag cgcatgacca acttcgacaa gaacctgccc
     1981 aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtacaacgag
     2041 ctgaccaagg tgaagtacgt gaccgagggc atgcgcaagc ccgccttcct gagcggcgag
     2101 cagaagaagg ccatcgtgga cctgctgttc aagaccaacc gcaaggtgac cgtgaagcag
     2161 ctgaaggagg actacttcaa gaagatcgag tgcttcgaca gcgtggagat cagcggcgtg
     2221 gaggaccgct tcaacgccag cctgggcacc taccacgacc tgctgaagat catcaaggac
     2281 aaggacttcc tggacaacga ggagaacgag gacatcctgg aggacatcgt gctgaccctg
     2341 accctgttcg aggaccgcga gatgatcgag gagcgcctga agacctacgc ccacctgttc
     2401 gacgacaagg tgatgaagca gctgaagcgc cgccgctaca ccggctgggg ccgcctgagc
     2461 cgcaagctta tcaacggcat ccgcgacaag cagagcggca agaccatcct ggacttcctg
     2521 aagagcgacg gcttcgccaa ccgcaacttc atgcagctga tccacgacga cagcctgacc
     2581 ttcaaggagg acatccagaa ggcccaggtg agcggccagg gcgacagcct gcacgagcac
     2641 atcgccaacc tggccggcag ccccgccatc aagaagggca tcctgcagac cgtgaaggtg
     2701 gtggacgagc tggtgaaggt gatgggccgc cacaagcccg agaacatcgt gatcgagatg
     2761 gcccgcgaga accagaccac ccagaagggc cagaagaaca gccgcgagcg catgaagcgc
     2821 atcgaggagg gcatcaagga gctgggcagc cagatcctga aggagcaccc cgtggagaac
     2881 acccagctgc agaacgagaa gctgtacctg tactacctgc agaacggccg cgacatgtac
     2941 gtggaccagg agctggacat caaccgcctg agcgactacg acgtggacca catcgtgccc
     3001 cagagcttcc tgaaggacga cagcatcgac aacaaggtgc tgacccgcag cgacaagaac
     3061 cgcggcaaga gcgacaacgt gcccagcgag gaggtggtga agaagatgaa gaactactgg
     3121 cgccagctgc tgaacgccaa gctgatcacc cagcgcaagt tcgacaacct gaccaaggcc
     3181 gagcgcggcg gcctgagcga gctggacaag gccggcttca tcaagcgcca gctggtggag
     3241 acccgccaga tcaccaagca cgtggcccag atcctggaca gccgcatgaa caccaagtac
     3301 gacgagaacg acaagctgat ccgcgaggtg aaggtgatca ccctgaagag caagctggtg
     3361 agcgacttcc gcaaggactt ccagttctac aaggtgcgcg agatcaacaa ctaccaccac
     3421 gcccacgacg cctacctgaa cgccgtggtg ggcaccgccc tgatcaagaa gtaccccaag
     3481 ctggagagcg agttcgtgta cggcgactac aaggtgtacg acgtgcgcaa gatgatcgcc
     3541 aagagcgagc aggagatcgg caaggccacc gccaagtact tcttctacag caacatcatg
     3601 aacttcttca agaccgagat caccctggcc aacggcgaga tccgcaagcg ccccctgatc
     3661 gagaccaacg gcgagaccgg cgagatcgtg tgggacaagg gccgcgactt cgccaccgtg
     3721 cgcaaggtgc tgagcatgcc ccaggtgaac atcgtgaaga agaccgaggt gcagaccggc
     3781 ggcttcagca aggagagcat cctgcccaag cgcaacagcg acaagctgat cgcccgcaag
     3841 aaggactggg accccaagaa gtacggcggc ttcgacagcc ccaccgtggc ctacagcgtg
     3901 ctggtggtgg ccaaggtgga gaagggcaag agcaagaagc tgaagagcgt gaaggagctg
     3961 ctgggcatca ccatcatgga gcgcagcagc ttcgagaaga accccatcga cttcctggag
     4021 gccaagggct acaaggaggt gaagaaggac ctgatcatca agctgcccaa gtacagcctg
     4081 ttcgagctgg agaacggccg caagcgcatg ctggccagcg ccggcgagct gcagaagggc
     4141 aacgagctgg ccctgcccag caagtacgtg aacttcctgt acctggccag ccactacgag
     4201 aagctgaagg gcagccccga ggacaacgag cagaagcagc tgttcgtgga gcagcacaag
     4261 cactacctgg acgagatcat cgagcagatc agcgagttca gcaagcgcgt gatcctggcc
     4321 gacgccaacc tggacaaggt gctgagcgcc tacaacaagc accgcgacaa gcccatccgc
     4381 gagcaggccg agaacatcat ccacctgttc accctgacca acctgggcgc ccccgccgcc
     4441 ttcaagtact tcgacaccac catcgaccgc aagcgctaca ccagcaccaa ggaggtgctg
     4501 gacgccaccc tgatccacca gagcatcacc ggtctgtacg agacccgcat cgacctgagc
     4561 cagctgggcg gcgacggcgg ctccggacct ccaaagaaaa agagaaaagt atacccctac
     4621 gacgtgcccg actacgccta acacccagct ttcttgtaca aagtggtgat atcccgcggc
     4681 catgctagag tccgcaaaaa tcaccagtct ctctctacaa atctatctct ctctattttt
     4741 ctccagaata atgtgtgagt agttcccaga taagggaatt agggttctta tagggtttcg
     4801 ctcatgtgtt gagcatataa gaaaccctta gtatgtattt gtatttgtaa aatacttcta
     4861 tcaataaaat ttctaattcc taaaaccaaa atccagtgac ctgcaggcat gcgacgtcgg
     4921 gcccaagctt agcttgagct tggatcagat tgtcgtttcc cgccttcagt ttaaactatc
     4981 agtgtttgac aggatatatt ggcgggtaaa cctaagagaa aagagcgttt attagaataa
     5041 tcggatattt aaaagggcgt gaaaaggttt atccgttcgt ccatttgtat gtgcatgcca
     5101 accacagggt tcccctcggg atcaaagtac tttaaagtac tttaaagtac tttaaagtac
     5161 tttgatccaa cccctccgct gctatagtgc agtcggcttc tgacgttcag tgcagccgtc
     5221 ttctgaaaac gacatgtcgc acaagtccta agttacgcga caggctgccg ccctgccctt
     5281 ttcctggcgt tttcttgtcg cgtgttttag tcgcataaag tagaatactt gcgactagaa
     5341 ccggagacat tacgccatga acaagagcgc cgccgctggc ctgctgggct atgcccgcgt
     5401 cagcaccgac gaccaggact tgaccaacca acgggccgaa ctgcacgcgg ccggctgcac
     5461 caagctgttt tccgagaaga tcaccggcac caggcgcgac cgcccggagc tggccaggat
     5521 gcttgaccac ctacgccctg gcgacgttgt gacagtgacc aggctagacc gcctggcccg
     5581 cagcacccgc gacctactgg acattgccga gcgcatccag gaggccggcg cgggcctgcg
     5641 tagcctggca gagccgtggg ccgacaccac cacgccggcc ggccgcatgg tgttgaccgt
     5701 gttcgccggc attgccgagt tcgagcgttc cctaatcatc gaccgcaccc ggagcgggcg
     5761 cgaggccgcc aaggcccgag gcgtgaagtt tggcccccgc cctaccctca ccccggcaca
     5821 gatcgcgcac gcccgcgagc tgatcgacca ggaaggccgc accgtgaaag aggcggctgc
     5881 actgcttggc gtgcatcgct cgaccctgta ccgcgcactt gagcgcagcg aggaagtgac
     5941 gcccaccgag gccaggcggc gcggtgcctt ccgtgaggac gcattgaccg aggccgacgc
     6001 cctggcggcc gccgagaatg aacgccaaga ggaacaagca tgaaaccgca ccaggacggc
     6061 caggacgaac cgtttttcat taccgaagag atcgaggcgg agatgatcgc ggccgggtac
     6121 gtgttcgagc cgcccgcgca cgtctcaacc gtgcggctgc atgaaatcct ggccggtttg
     6181 tctgatgcca agctggcggc ctggccggcc agcttggccg ctgaagaaac cgagcgccgc
     6241 cgtctaaaaa ggtgatgtgt atttgagtaa aacagcttgc gtcatgcggt cgctgcgtat
     6301 atgatgcgat gagtaaataa acaaatacgc aaggggaacg catgaaggtt atcgctgtac
     6361 ttaaccagaa aggcgggtca ggcaagacga ccatcgcaac ccatctagcc cgcgccctgc
     6421 aactcgccgg ggccgatgtt ctgttagtcg attccgatcc ccagggcagt gcccgcgatt
     6481 gggcggccgt gcgggaagat caaccgctaa ccgttgtcgg catcgaccgc ccgacgattg
     6541 accgcgacgt gaaggccatc ggccggcgcg acttcgtagt gatcgacgga gcgccccagg
     6601 cggcggactt ggctgtgtcc gcgatcaagg cagccgactt cgtgctgatt ccggtgcagc
     6661 caagccctta cgacatatgg gccaccgccg acctggtgga gctggttaag cagcgcattg
     6721 aggtcacgga tggaaggcta caagcggcct ttgtcgtgtc gcgggcgatc aaaggcacgc
     6781 gcatcggcgg tgaggttgcc gaggcgctgg ccgggtacga gctgcccatt cttgagtccc
     6841 gtatcacgca gcgcgtgagc tacccaggca ctgccgccgc cggcacaacc gttcttgaat
     6901 cagaacccga gggcgacgct gcccgcgagg tccaggcgct ggccgctgaa attaaatcaa
     6961 aactcatttg agttaatgag gtaaagagaa aatgagcaaa agcacaaaca cgctaagtgc
     7021 cggccgtccg agcgcacgca gcagcaaggc tgcaacgttg gccagcctgg cagacacgcc
     7081 agccatgaag cgggtcaact ttcagttgcc ggcggaggat cacaccaagc tgaagatgta
     7141 cgcggtacgc caaggcaaga ccattaccga gctgctatct gaatacatcg cgcagctacc
     7201 agagtaaatg agcaaatgaa taaatgagta gatgaatttt agcggctaaa ggaggcggca
     7261 tggaaaatca agaacaacca ggcaccgacg ccgtggaatg ccccatgtgt ggaggaacgg
     7321 gcggttggcc aggcgtaagc ggctgggttg tctgccggcc ctgcaatggc actggaaccc
     7381 ccaagcccga ggaatcggcg tgagcggtcg caaaccatcc ggcccggtac aaatcggcgc
     7441 ggcgctgggt gatgacctgg tggagaagtt gaaggccgcg caggccgccc agcggcaacg
     7501 catcgaggca gaagcacgcc ccggtgaatc gtggcaagcg gccgctgatc gaatccgcaa
     7561 agaatcccgg caaccgccgg cagccggtgc gccgtcgatt aggaagccgc ccaagggcga
     7621 cgagcaacca gattttttcg ttccgatgct ctatgacgtg ggcacccgcg atagtcgcag
     7681 catcatggac gtggccgttt tccgtctgtc gaagcgtgac cgacgagctg gcgaggtgat
     7741 ccgctacgag cttccagacg ggcacgtaga ggtttccgca gggccggccg gcatggccag
     7801 tgtgtgggat tacgacctgg tactgatggc ggtttcccat ctaaccgaat ccatgaaccg
     7861 ataccgggaa gggaagggag acaagcccgg ccgcgtgttc cgtccacacg ttgcggacgt
     7921 actcaagttc tgccggcgag ccgatggcgg aaagcagaaa gacgacctgg tagaaacctg
     7981 cattcggtta aacaccacgc acgttgccat gcagcgtacg aagaaggcca agaacggccg
     8041 cctggtgacg gtatccgagg gtgaagcctt gattagccgc tacaagatcg taaagagcga
     8101 aaccgggcgg ccggagtaca tcgagatcga gctagctgat tggatgtacc gcgagatcac
     8161 agaaggcaag aacccggacg tgctgacggt tcaccccgat tactttttga tcgatcccgg
     8221 catcggccgt tttctctacc gcctggcacg ccgcgccgca ggcaaggcag aagccagatg
     8281 gttgttcaag acgatctacg aacgcagtgg cagcgccgga gagttcaaga agttctgttt
     8341 caccgtgcgc aagctgatcg ggtcaaatga cctgccggag tacgatttga aggaggaggc
     8401 ggggcaggct ggcccgatcc tagtcatgcg ctaccgcaac ctgatcgagg gcgaagcatc
     8461 cgccggttcc taatgtacgg agcagatgct agggcaaatt gccctagcag gggaaaaagg
     8521 tcgaaaaggt ctctttcctg tggatagcac gtacattggg aacccaaagc cgtacattgg
     8581 gaaccggaac ccgtacattg ggaacccaaa gccgtacatt gggaaccggt cacacatgta
     8641 agtgactgat ataaaagaga aaaaaggcga tttttccgcc taaaactctt taaaacttat
     8701 taaaactctt aaaacccgcc tggcctgtgc ataactgtct ggccagcgca cagccgaaga
     8761 gctgcaaaaa gcgcctaccc ttcggtcgct gcgctcccta cgccccgccg cttcgcgtcg
     8821 gcctatcgcg gccgctggcc gctcaaaaat ggctggccta cggccaggca atctaccagg
     8881 gcgcggacaa gccgcgccgt cgccactcga ccgccggcgc ccacatcaag gcaccctgcc
     8941 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     9001 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     9061 ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg
     9121 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     9181 accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac
     9241 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     9301 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     9361 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     9421 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     9481 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     9541 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     9601 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     9661 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     9721 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     9781 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     9841 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     9901 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     9961 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
    10021 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgcatg atatatctcc
    10081 caatttgtgt agggcttatt atgcacgctt aaaaataata aaagcagact tgacctgata
    10141 gtttggctgt gagcaattat gtgcttagtg catctaacgc ttgagttaag ccgcgccgcg
    10201 aagcggcgtc ggcttgaacg aatttctagc tagacattat ttgccgacta ccttggtgat
    10261 ctcgcctttc acgtagtgga caaattcttc caactgatct gcgcgcgagg ccaagcgatc
    10321 ttcttcttgt ccaagataag cctgtctagc ttcaagtatg acgggctgat actgggccgg
    10381 caggcgctcc attgcccagt cggcagcgac atccttcggc gcgattttgc cggttactgc
    10441 gctgtaccaa atgcgggaca acgtaagcac tacatttcgc tcatcgccag cccagtcggg
    10501 cggcgagttc catagcgtta aggtttcatt tagcgcctca aatagatcct gttcaggaac
    10561 cggatcaaag agttcctccg ccgctggacc taccaaggca acgctatgtt ctcttgcttt
    10621 tgtcagcaag atagccagat caatgtcgat cgtggctggc tcgaagatac ctgcaagaat
    10681 gtcattgcgc tgccattctc caaattgcag ttcgcgctta gctggataac gccacggaat
    10741 gatgtcgtcg tgcacaacaa tggtgacttc tacagcgcgg agaatctcgc tctctccagg
    10801 ggaagccgaa gtttccaaaa ggtcgttgat caaagctcgc cgcgttgttt catcaagcct
    10861 tacggtcacc gtaaccagca aatcaatatc actgtgtggc ttcaggccgc catccactgc
    10921 ggagccgtac aaatgtacgg ccagcaacgt cggttcgaga tggcgctcga tgacgccaac
    10981 tacctctgat agttgagtcg atacttcggc gatcaccgct tcccccatga tgtttaactt
    11041 tgttttaggg cgactgccct gctgcgtaac atcgttgctg ctccataaca tcaaacatcg
    11101 acccacggcg taacgcgctt gctgcttgga tgcccgaggc atagactgta ccccaaaaaa
    11161 acagtcataa caagccatga aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa
    11221 ggttctggac cagttgcgtg agcgcatacg ctacttgcat tacagcttac gaaccgaaca
    11281 ggcttatgtc cactgggttc gtgcccgaat tgatcacagg cagcaacgct ctgtcatcgt
    11341 tacaatcaac atgctaccct ccgcgagatc atccgtgttt caaacccggc agcttagttg
    11401 ccgttcttcc gaatagcatc ggtaacatga gcaaagtctg ccgccttaca acggctctcc
    11461 cgctgacgcc gtcccggact gatgggctgc ctgtatcgag tggtgatttt gtgccgagct
    11521 gccggtcggg gagctgttgg ctggctggtg gcaggatata ttgtggtgta aacaaattga
    11581 cgcttagaca acttaataac acattgcgga cgtttttaat gtactgaatt aacgccgaat
    11641 tgaattatca gcttgcatgc cggtcgatct agtaacatag atgacaccgc gcgcgataat
    11701 ttatcctagt ttgcgcgcta tattttgttt tctatcgcgt attaaatgta taattgcggg
    11761 actctaatca taaaaaccca tctcataaat aacgtcatgc attacatgtt aattattaca
    11821 tgcttaacgt aattcaacag aaattatatg ataatcatcg caagaccggc aacaggattc
    11881 aatcttaaga aactttattg ccaaatgttt gaacgatctg cttgactcta gggaattaat
    11941 tcctgaatca ctgcgaccgg ccctcccgcg acccagccga gcgagcttag cgaactgtgg
    12001 acgagaactg tgccaccaag cgtaaggccg ttctctcgca tttgccttgc taggctcgcg
    12061 cgagttgctg gctgaggtca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc
    12121 gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc
    12181 tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc
    12241 cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag
    12301 gcatcgccat gggtcacgac gagatcatcg ccgtcgggca tgcgcgcctt gagcctggcg
    12361 aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga
    12421 ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg
    12481 caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc
    12541 tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc
    12601 cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg
    12661 gccagccacg atagccgcgc tgcctcgtcc tgcagttcat tcagggcacc ggacaggtcg
    12721 gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag
    12781 cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga
    12841 gaacctgcgt gcaatccatc ttgttcaatc atagggggga ttcgagttga gagtgaatat
    12901 gagactctaa ttggataccg aggggaattt atggaacgtc agtggagcat ttttgacaag
    12961 aaatatttgc tagctgatag tgaccttagg cgacttttga acgcgcaata atggtttctg
    13021 acgtatgtgc ttagctcatt aaactccaga aacccgcggc tcagtggctc cttcaacgtt
    13081 gcggttctgt cagttccaaa cgtaaaacgg cttgtcccgc gtcatcggcg ggggtcataa
    13141 cgtgactccc ttaattctcc gctcatgatc gataattcga gctcaaaaaa gcaccgactc
    13201 ggtgccactt tttcaagttg ataacggact agccttattt taacttgcta tttctagctc
    13261 taaaacctct gcaggtgact ctcgagacac ctgcctccaa tcactacttc gactctagct
    13321 gtatataaac tcagcttcgt tttcttatct aagcgatgtg ggacttttga agattgtttt
    13381 caacttaaat gggcctatat aagaaatact attgttcttt cccatataaa tgggcctgct
    13441 tctcttcttt cagattccca ggggcctttt gaagattatc ttcatatctt aagaatgaag
    13501 atgttttatt caatcaaatt cttgaaggtt cgatgcctaa tcattcgaat tcgagctctc
    13561 ccatatggtc gactagagcc aagctgatct cctttgcccc ggagatcacc atggacgact
    13621 ttctctatct ctacgatcta ggaagaaagt tcgacggaga aggtgacgat accatgttca
    13681 ccaccgataa tgagaagatt agcctcttca atttcagaaa gaatgctgac ccacagatgg
    13741 ttagagaggc ctacgcggca ggtctcatca agacgatcta cccgagtaat aatctccagg
    13801 agatcaaata ccttcccaag aaggttaaag atgcagtcaa aagattcagg actaactgca
    13861 tcaagaacac agagaaagat atatttctca agatcagaag tactattcca gtatggacga
    13921 ttcaaggctt gcttcataaa ccaaggcaag taatagagat tggagtctct aagaaagtag
    13981 ttcctactga atcaaaggcc atggagtcaa aaattcagat cgaggatcta acagaactcg
    14041 ccgtgaagac tggcgaacag ttcatacaga gtcttttacg actcaatgac aagaagaaaa
    14101 tcttcgtcaa catggtggag cacgacactc tcgtctactc caagaatatc aaagatacag
    14161 tctcagaaga ccaaagggct at
//
