LOCUS       sequence-395472-       17011 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR/Cas based plant genome editing and gene regulation; expresses
            3×FLAG-NLS-zCas9-NLS, gRNA scaffold for insertion of target sequence
            (OsU3 promoter), Bar resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17011)
  AUTHORS   Xing HL, Dong L, Wang ZP, Zhang HY, Han CY, Liu B, Wang XC, Chen QJ
  TITLE     A CRISPR/Cas9 toolkit for multiplex genome editing in plants.
  JOURNAL   BMC Plant Biol. 2014 Nov 29;14(1):327.
  PUBMED    25432517
REFERENCE   2  (bases 1 to 17011)
  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: "BMC Plant 
            Biol."; date: "2014-11-29"; volume: "14(1)"; pages: "327"
COMMENT     Sequence Label: pBUE411
FEATURES             Location/Qualifiers
     source          1..17011
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        84..463
                     /label=OsU3 promoter
                     /note="Oryza sativa (rice) snRNA U3 promoter"
     CDS             743..1534
                     /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_RNA        1686..1761
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2065..4053
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     CDS             4072..4137
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4144..4164
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4189..8289
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4189..8289
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     RBS             8202..8210
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             8290..8337
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(8994..9010)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8994..9010)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9007..9029)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9018..9034
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(9042..9048,9049..9066,9067..9072))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9087..9108
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        9299..9644
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     9587..9607
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             9695..10246
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     polyA_signal    10253..10427
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    10505..10529
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             10954..11748
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      11835..12423
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12324..12343
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12577..12594
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    12609..12749
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(12675..12694)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     12835..12857
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      13093..13287
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(13353..14426)
                     /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"
     CDS             complement(14855..15484)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    16785..16809
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     join(16997..17011,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(17011,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgcc aagcttagta attcatccag gtctccaagt tctaggattt
       61 tcagaactgc aacttatttt atcaaggaat ctttaaacat acgaacagat cacttaaagt
      121 tcttctgaag caacttaaag ttatcaggca tgcatggatc ttggaggaat cagatgtgca
      181 gtcagggacc atagcacaag acaggcgtct tctactggtg ctaccagcaa atgctggaag
      241 ccgggaacac tgggtacgtt ggaaaccacg tgatgtgaag aagtaagata aactgtagga
      301 gaaaagcatt tcgtagtggg ccatgaagcc tttcaggaca tgtattgcag tatgggccgg
      361 cccattacgc aattggacga caacaaagac tagtattagt accacctcgg ctatccacat
      421 agatcaaagc tgatttaaaa gagttgtgca gatgatccgt ggcgtgagac caacccagtg
      481 gacataagcc tgttcggttc gtaagctgta atgcaagtag cgtatgcgct cacgcaactg
      541 gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt tcatggcttg
      601 ttatgactgt ttttttgggg tacagtctat gcctcgggca tccaagcagc aagcgcgtta
      661 cgccgtgggt cgatgtttga tgttatggag cagcaacgat gttacgcagc agggcagtcg
      721 ccctaaaaca aagttaaaca tcatggggga agcggtgatc gccgaagtat cgactcaact
      781 atcagaggta gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt
      841 gtacggctcc gcagtggatg gcggcctgaa gccacacagt gatattgatt tgctggttac
      901 ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg atcaacgacc ttttggaaac
      961 ttcggcttcc cctggagaga gcgagattct ccgcgctgta gaagtcacca ttgttgtgca
     1021 cgacgacatc attccgtggc gttatccagc taagcgcgaa ctgcaatttg gagaatggca
     1081 gcgcaatgac attcttgcag gtatcttcga gccagccacg atcgacattg atctggctat
     1141 cttgctgaca aaagcaagag aacatagcgt tgccttggta ggtccagcgg cggaggaact
     1201 ctttgatccg gttcctgaac aggatctatt tgaggcgcta aatgaaacct taacgctatg
     1261 gaactcgccg cccgactggg ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat
     1321 ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat
     1381 ggagcgcctg ccggcccagt atcagcccgt catacttgaa gctagacagg cttatcttgg
     1441 acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg tccactacgt
     1501 gaaaggcgag atcaccaagg tagtcggcaa ataatgtcta gctagaaatt cgttcaagcc
     1561 gacgccgctt cgcggcgcgg cttaactcaa gcgttagatg cactaagcac ataattgctc
     1621 acagccaaac tatcaggtca agtctgcttt tattattttt aagcgtgcat aataagccgg
     1681 tctcggtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa
     1741 aaagtggcac cgagtcggtg cttttttttt tcgttttgca ttgagttttc tccgtcgcat
     1801 gtttgcagtt ttattttccg ttttgcattg aaatttctcc gtctcatgtt tgcagcgtgt
     1861 tcaaaaagta cgcagctgta tttcacttat ttacggcgcc acattttcat gccgtttgtg
     1921 ccaactatcc cgagctagtg aatacagctt ggcttcacac aacactggtg acccgctgac
     1981 ctgctcgtac ctcgtaccgt cgtacggcac agcatttgga attaaagggt gtgatcgata
     2041 ctgcttgctg ctaagcttgc atgcctgcag tgcagcgtga cccggtcgtg cccctctcta
     2101 gagataatga gcattgcatg tctaagttat aaaaaattac cacatatttt ttttgtcaca
     2161 cttgtttgaa gtgcagttta tctatcttta tacatatatt taaactttac tctacgaata
     2221 atataatcta tagtactaca ataatatcag tgttttagag aatcatataa atgaacagtt
     2281 agacatggtc taaaggacaa ttgagtattt tgacaacagg actctacagt tttatctttt
     2341 tagtgtgcat gtgttctcct ttttttttgc aaatagcttc acctatataa tacttcatcc
     2401 attttattag tacatccatt tagggtttag ggttaatggt ttttatagac taattttttt
     2461 agtacatcta ttttattcta ttttagcctc taaattaaga aaactaaaac tctattttag
     2521 tttttttatt taataattta gatataaaat agaataaaat aaagtgacta aaaattaaac
     2581 aaataccctt taagaaatta aaaaaactaa ggaaacattt ttcttgtttc gagtagataa
     2641 tgccagcctg ttaaacgccg tcgacgagtc taacggacac caaccagcga accagcagcg
     2701 tcgcgtcggg ccaagcgaag cagacggcac ggcatctctg tcgctgcctc tggacccctc
     2761 tcgagagttc cgctccaccg ttggacttgc tccgctgtcg gcatccagaa attgcgtggc
     2821 ggagcggcag acgtgagccg gcacggcagg cggcctcctc ctcctctcac ggcaccggca
     2881 gctacggggg attcctttcc caccgctcct tcgctttccc ttcctcgccn gccgtaataa
     2941 atagacaccc cctccacacc ctctttcccc aacctcgtgt tgttcggagc gcacacacac
     3001 acaaccagat ctcccccaaa tccacccgtc ggcacctccg cttcaaggta cgccgctcgt
     3061 cctccccccc cctctctacc ttctctagat cggcgttccg gtccatggtt agggcccggt
     3121 agttctactt ctgttcatgt ttgtgttaga tccgtgtttg tgttagatcc gtgctgctag
     3181 cgttcgtaca cggatgcgac ctgtacgtca gacacgttct gattgctaac ttgccagtgt
     3241 ttctctttgg ggaatcctgg gatggctcta gccgttccgc agacgggatc gatttcatga
     3301 ttttttttgt ttcgttgcat agggtttggt ttgccctttt cctttatttc aatatatgcc
     3361 gtgcacttgt ttgtcgggtc atcttttcat gctttttttt gtcttggttg tgatgatgtg
     3421 gtctggttgg gcggtcgttc tagatcggag tagaattctg tttcaaacta cctggtggat
     3481 ttattaattt tggatctgta tgtgtgtgcc atacatattc atagttacga attgaagatg
     3541 atggatggaa atatcgatct aggataggta tacatgttga tgcgggtttt actgatgcat
     3601 atacagagat gctttttgtt cgcttggttg tgatgatgtg gtgtggttgg gcggtcgttc
     3661 attcgttcta gatcggagta gaatactgtt tcaaactacc tggtgtattt attaattttg
     3721 gaactgtatg tgtgtgtcat acatcttcat agttacgagt ttaagatgga tggaaatatc
     3781 gatctaggat aggtatacat gttgatgtgg gttttactga tgcatataca tgatggcata
     3841 tgcagcatct attcatatgc tctaaccttg agtacctatc tattataata aacaagtatg
     3901 ttttataatt attttgatct tgatatactt ggatgatggc atatgcagca gctatatgtg
     3961 gattttttta gccctgcctt catacgctat ttatttgctt ggtactgttt cttttgtcga
     4021 tgctcaccct gttgtttggt gttacttctg cagccctagg cctactagat ggattacaag
     4081 gaccacgacg gggattacaa ggaccacgac attgattaca aggatgatga tgacaagatg
     4141 gctccgaaga agaagaggaa ggttggcatc cacggggtgc cagctgctga caagaagtac
     4201 tcgatcggcc tcgatattgg gactaactct gttggctggg ccgtgatcac cgacgagtac
     4261 aaggtgccct caaagaagtt caaggtcctg ggcaacaccg atcggcattc catcaagaag
     4321 aatctcattg gcgctctcct gttcgacagc ggcgagacgg ctgaggctac gcggctcaag
     4381 cgcaccgccc gcaggcggta cacgcgcagg aagaatcgca tctgctacct gcaggagatt
     4441 ttctccaacg agatggcgaa ggttgacgat tctttcttcc acaggctgga ggagtcattc
     4501 ctcgtggagg aggataagaa gcacgagcgg catccaatct tcggcaacat tgtcgacgag
     4561 gttgcctacc acgagaagta ccctacgatc taccatctgc ggaagaagct cgtggactcc
     4621 acagataagg cggacctccg cctgatctac ctcgctctgg cccacatgat taagttcagg
     4681 ggccatttcc tgatcgaggg ggatctcaac ccggacaata gcgatgttga caagctgttc
     4741 atccagctcg tgcagacgta caaccagctc ttcgaggaga accccattaa tgcgtcaggc
     4801 gtcgacgcga aggctatcct gtccgctagg ctctcgaagt ctcggcgcct cgagaacctg
     4861 atcgcccagc tgccgggcga gaagaagaac ggcctgttcg ggaatctcat tgcgctcagc
     4921 ctggggctca cgcccaactt caagtcgaat ttcgatctcg ctgaggacgc caagctgcag
     4981 ctctccaagg acacatacga cgatgacctg gataacctcc tggcccagat cggcgatcag
     5041 tacgcggacc tgttcctcgc tgccaagaat ctgtcggacg ccatcctcct gtctgatatt
     5101 ctcagggtga acaccgagat tacgaaggct ccgctctcag cctccatgat caagcgctac
     5161 gacgagcacc atcaggatct gaccctcctg aaggcgctgg tcaggcagca gctccccgag
     5221 aagtacaagg agatcttctt cgatcagtcg aagaacggct acgctgggta cattgacggc
     5281 ggggcctctc aggaggagtt ctacaagttc atcaagccga ttctggagaa gatggacggc
     5341 acggaggagc tgctggtgaa gctcaatcgc gaggacctcc tgaggaagca gcggacattc
     5401 gataacggca gcatcccaca ccagattcat ctcggggagc tgcacgctat cctgaggagg
     5461 caggaggact tctacccttt cctcaaggat aaccgcgaga agatcgagaa gattctgact
     5521 ttcaggatcc cgtactacgt cggcccactc gctaggggca actcccgctt cgcttggatg
     5581 acccgcaagt cagaggagac gatcacgccg tggaacttcg aggaggtggt cgacaagggc
     5641 gctagcgctc agtcgttcat cgagaggatg acgaatttcg acaagaacct gccaaatgag
     5701 aaggtgctcc ctaagcactc gctcctgtac gagtacttca cagtctacaa cgagctgact
     5761 aaggtgaagt atgtgaccga gggcatgagg aagccggctt tcctgtctgg ggagcagaag
     5821 aaggccatcg tggacctcct gttcaagacc aaccggaagg tcacggttaa gcagctcaag
     5881 gaggactact tcaagaagat tgagtgcttc gattcggtcg agatctctgg cgttgaggac
     5941 cgcttcaacg cctccctggg gacctaccac gatctcctga agatcattaa ggataaggac
     6001 ttcctggaca acgaggagaa tgaggatatc ctcgaggaca ttgtgctgac actcactctg
     6061 ttcgaggacc gggagatgat cgaggagcgc ctgaagactt acgcccatct cttcgatgac
     6121 aaggtcatga agcagctcaa gaggaggagg tacaccggct gggggaggct gagcaggaag
     6181 ctcatcaacg gcattcggga caagcagtcc gggaagacga tcctcgactt cctgaagagc
     6241 gatggcttcg cgaaccgcaa tttcatgcag ctgattcacg atgacagcct cacattcaag
     6301 gaggatatcc agaaggctca ggtgagcggc cagggggact cgctgcacga gcatatcgcg
     6361 aacctcgctg gctcgccagc tatcaagaag gggattctgc agaccgtgaa ggttgtggac
     6421 gagctggtga aggtcatggg caggcacaag cctgagaaca tcgtcattga gatggcccgg
     6481 gagaatcaga ccacgcagaa gggccagaag aactcacgcg agaggatgaa gaggatcgag
     6541 gagggcatta aggagctggg gtcccagatc ctcaaggagc acccggtgga gaacacgcag
     6601 ctgcagaatg agaagctcta cctgtactac ctccagaatg gccgcgatat gtatgtggac
     6661 caggagctgg atattaacag gctcagcgat tacgacgtcg atcatatcgt tccacagtca
     6721 ttcctgaagg atgactccat tgacaacaag gtcctcacca ggtcggacaa gaaccggggc
     6781 aagtctgata atgttccttc agaggaggtc gttaagaaga tgaagaacta ctggcgccag
     6841 ctcctgaatg ccaagctgat cacgcagcgg aagttcgata acctcacaaa ggctgagagg
     6901 ggcgggctct ctgagctgga caaggcgggc ttcatcaaga ggcagctggt cgagacacgg
     6961 cagatcacta agcacgttgc gcagattctc gactcacgga tgaacactaa gtacgatgag
     7021 aatgacaagc tgatccgcga ggtgaaggtc atcaccctga agtcaaagct cgtctccgac
     7081 ttcaggaagg atttccagtt ctacaaggtt cgggagatca acaattacca ccatgcccat
     7141 gacgcgtacc tgaacgcggt ggtcggcaca gctctgatca agaagtaccc aaagctcgag
     7201 agcgagttcg tgtacgggga ctacaaggtt tacgatgtga ggaagatgat cgccaagtcg
     7261 gagcaggaga ttggcaaggc taccgccaag tacttcttct actctaacat tatgaatttc
     7321 ttcaagacag agatcactct ggccaatggc gagatccgga agcgccccct catcgagacg
     7381 aacggcgaga cgggggagat cgtgtgggac aagggcaggg atttcgcgac cgtcaggaag
     7441 gttctctcca tgccacaagt gaatatcgtc aagaagacag aggtccagac tggcgggttc
     7501 tctaaggagt caattctgcc taagcggaac agcgacaagc tcatcgcccg caagaaggac
     7561 tgggatccga agaagtacgg cgggttcgac agccccactg tggcctactc ggtcctggtt
     7621 gtggcgaagg ttgagaaggg caagtccaag aagctcaaga gcgtgaagga gctgctgggg
     7681 atcacgatta tggagcgctc cagcttcgag aagaacccga tcgatttcct ggaggcgaag
     7741 ggctacaagg aggtgaagaa ggacctgatc attaagctcc ccaagtactc actcttcgag
     7801 ctggagaacg gcaggaagcg gatgctggct tccgctggcg agctgcagaa ggggaacgag
     7861 ctggctctgc cgtccaagta tgtgaacttc ctctacctgg cctcccacta cgagaagctc
     7921 aagggcagcc ccgaggacaa cgagcagaag cagctgttcg tcgagcagca caagcattac
     7981 ctcgacgaga tcattgagca gatttccgag ttctccaagc gcgtgatcct ggccgacgcg
     8041 aatctggata aggtcctctc cgcgtacaac aagcaccgcg acaagccaat cagggagcag
     8101 gctgagaata tcattcatct cttcaccctg acgaacctcg gcgcccctgc tgctttcaag
     8161 tacttcgaca caactatcga tcgcaagagg tacacaagca ctaaggaggt cctggacgcg
     8221 accctcatcc accagtcgat taccggcctc tacgagacgc gcatcgacct gtctcagctc
     8281 gggggcgaca agcggccagc ggcgacgaag aaggcggggc aggcgaagaa gaagaagtga
     8341 gctcagagct ttcgttcgta tcatcggttt cgacaacgtt cgtcaagttc aatgcatcag
     8401 tttcattgcg cacacaccag aatcctactg agtttgagta ttatggcatt gggaaaactg
     8461 tttttcttgt accatttgtt gtgcttgtaa tttactgtgt tttttattcg gttttcgcta
     8521 tcgaactgtg aaatggaaat ggatggagaa gagttaatga atgatatggt ccttttgttc
     8581 attctcaaat taatattatt tgttttttct cttatttgtt gtgtgttgaa tttgaaatta
     8641 taagagatat gcaaacattt tgttttgagt aaaaatgtgt caaatcgtgg cctctaatga
     8701 ccgaagttaa tatgaggagt aaaacacttg tagttgtacc attatgctta ttcactaggc
     8761 aacaaatata ttttcagacc tagaaaagct gcaaatgtta ctgaatacaa gtatgtcctc
     8821 ttgtgtttta gacatttatg aactttcctt tatgtaattt tccagaatcc ttgtcagatt
     8881 ctaatcattg ctttataatt atagttatac tcatggattt gtagttgagt atgaaaatat
     8941 tttttaatgc attttatgac ttgccaattg attgacaacg aattcgtaat catgtcatag
     9001 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     9061 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     9121 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     9181 cgcgcgggga gaggcggttt gcgtattggc tagagcagct tgccaacatg gtggagcacg
     9241 acactctcgt ctactccaag aatatcaaag atacagtctc agaagaccaa agggctattg
     9301 agacttttca acaaagggta atatcgggaa acctcctcgg attccattgc ccagctatct
     9361 gtcacttcat caaaaggaca gtagaaaagg aaggtggcac ctacaaatgc catcattgcg
     9421 ataaaggaaa ggctatcgtt caagatgcct ctgccgacag tggtcccaaa gatggacccc
     9481 cacccacgag gagcatcgtg gaaaaagaag acgttccaac cacgtcttca aagcaagtgg
     9541 attgatgtga tatctccact gacgtaaggg atgacgcaca atcccactat ccttcgcaag
     9601 acccttcctc tatataagga agttcatttc atttggagag gacacgctga aatcaccagt
     9661 ctctctctac aaatctatct ctctcgagtc taccatgagc ccagaacgac gcccggccga
     9721 catccgccgt gccaccgagg cggacatgcc ggcggtctgc accatcgtca accactacat
     9781 cgagacaagc acggtcaact tccgtaccga gccgcaggaa ccgcaggagt ggacggacga
     9841 cctcgtccgt ctgcgggagc gctatccctg gctcgtcgcc gaggtggacg gcgaggtcgc
     9901 cggcatcgcc tacgcgggcc cctggaaggc acgcaacgcc tacgactgga cggccgagtc
     9961 gaccgtgtac gtctcccccc gccaccagcg gacgggactg ggctccacgc tctacaccca
    10021 cctgctgaag tccctggagg cacagggctt caagagcgtg gtcgctgtca tcgggctgcc
    10081 caacgacccg agcgtgcgca tgcacgaggc gctcggatat gccccccgcg gcatgctgcg
    10141 ggcggccggc ttcaagcacg ggaactggca tgacgtgggt ttctggcagc tggacttcag
    10201 cctgccggta ccgccccgtc cggtcctgcc cgtcaccgag atttgactcg agtttctcca
    10261 taataatgtg tgagtagttc ccagataagg gaattagggt tcctataggg tttcgctcat
    10321 gtgttgagca tataagaaac ccttagtatg tatttgtatt tgtaaaatac ttctatcaat
    10381 aaaatttcta attcctaaaa ccaaaatcca gtactaaaat ccagatcccc cgaattaatt
    10441 cggcgttaat tcagtacatt aaaaacgtcc gcaatgtgtt attaagttgt ctaagcgtca
    10501 atttgtttac accacaatat atcctgccac cagccagcca acagctcccc gaccggcagc
    10561 tcggcacaaa atcaccactc gatacaggca gcccatcagt ccgggacggc gtcagcggga
    10621 gagccgttgt aaggcggcag actttgctca tgttaccgat gctattcgga agaacggcaa
    10681 ctaagctgcc gggtttgaaa cacggatgat ctcgcggagg gtagcatgtt gattgtaacg
    10741 atgacagagc gttgctgcct gtgatcaccg cggtttcaaa atcggctccg tcgatactat
    10801 gttatacgcc aactttgaaa acaactttga aaaagctgtt ttctggtatt taaggtttta
    10861 gaatgcaagg aacagtgaat tggagttcgt cttgttataa ttagcttctt ggggtatctt
    10921 taaatactgt agaaaagagg aaggaaataa taaatggcta aaatgagaat atcaccggaa
    10981 ttgaaaaaac tgatcgaaaa ataccgctgc gtaaaagata cggaaggaat gtctcctgct
    11041 aaggtatata agctggtggg agaaaatgaa aacctatatt taaaaatgac ggacagccgg
    11101 tataaaggga ccacctatga tgtggaacgg gaaaaggaca tgatgctatg gctggaagga
    11161 aagctgcctg ttccaaaggt cctgcacttt gaacggcatg atggctggag caatctgctc
    11221 atgagtgagg ccgatggcgt cctttgctcg gaagagtatg aagatgaaca aagccctgaa
    11281 aagattatcg agctgtatgc ggagtgcatc aggctctttc actccatcga catatcggat
    11341 tgtccctata cgaatagctt agacagccgc ttagccgaat tggattactt actgaataac
    11401 gatctggccg atgtggattg cgaaaactgg gaagaagaca ctccatttaa agatccgcgc
    11461 gagctgtatg attttttaaa gacggaaaag cccgaagagg aacttgtctt ttcccacggc
    11521 gacctgggag acagcaacat ctttgtgaaa gatggcaaag taagtggctt tattgatctt
    11581 gggagaagcg gcagggcgga caagtggtat gacattgcct tctgcgtccg gtcgatcagg
    11641 gaggatatcg gggaagaaca gtatgtcgag ctattttttg acttactggg gatcaagcct
    11701 gattgggaga aaataaaata ttatatttta ctggatgaat tgttttagta cctagaatgc
    11761 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
    11821 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
    11881 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
    11941 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
    12001 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
    12061 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
    12121 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
    12181 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
    12241 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
    12301 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
    12361 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
    12421 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
    12481 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
    12541 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
    12601 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
    12661 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
    12721 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
    12781 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
    12841 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ggtgccttga
    12901 tgtgggcgcc ggcggtcgag tggcgacggc gcggcttgtc cgcgccctgg tagattgcct
    12961 ggccgtaggc cagccatttt tgagcggcca gcggccgcga taggccgacg cgaagcggcg
    13021 gggcgtaggg agcgcagcga ccgaagggta ggcgcttttt gcagctcttc ggctgtgcgc
    13081 tggccagaca gttatgcaca ggccaggcgg gttttaagag ttttaataag ttttaaagag
    13141 ttttaggcgg aaaaatcgcc ttttttctct tttatatcag tcacttacat gtgtgaccgg
    13201 ttcccaatgt acggctttgg gttcccaatg tacgggttcc ggttcccaat gtacggcttt
    13261 gggttcccaa tgtacgtgct atccacagga aacagacctt ttcgaccttt ttcccctgct
    13321 agggcaattt gccctagcat ctgctccgta cattaggaac cggcggatgc ttcgccctcg
    13381 atcaggttgc ggtagcgcat gactaggatc gggccagcct gccccgcctc ctccttcaaa
    13441 tcgtactccg gcaggtcatt tgacccgatc agcttgcgca cggtgaaaca gaacttcttg
    13501 aactctccgg cgctgccact gcgttcgtag atcgtcttga acaaccatct ggcttctgcc
    13561 ttgcctgcgg cgcggcgtgc caggcggtag agaaaacggc cgatgccggg atcgatcaaa
    13621 aagtaatcgg ggtgaaccgt cagcacgtcc gggttcttgc cttctgtgat ctcgcggtac
    13681 atccaatcag ctagctcgat ctcgatgtac tccggccgcc cggtttcgct ctttacgatc
    13741 ttgtagcggc taatcaaggc ttcaccctcg gataccgtca ccaggcggcc gttcttggcc
    13801 ttcttcgtac gctgcatggc aacgtgcgtg gtgtttaacc gaatgcaggt ttctaccagg
    13861 tcgtctttct gctttccgcc atcggctcgc cggcagaact tgagtacgtc cgcaacgtgt
    13921 ggacggaaca cgcggccggg cttgtctccc ttcccttccc ggtatcggtt catggattcg
    13981 gttagatggg aaaccgccat cagtaccagg tcgtaatccc acacactggc catgccggcc
    14041 ggccctgcgg aaacctctac gtgcccgtct ggaagctcgt agcggatcac ctcgccagct
    14101 cgtcggtcac gcttcgacag acggaaaacg gccacgtcca tgatgctgcg actatcgcgg
    14161 gtgcccacgt catagagcat cggaacgaaa aaatctggtt gctcgtcgcc cttgggcggc
    14221 ttcctaatcg acggcgcacc ggctgccggc ggttgccggg attctttgcg gattcgatca
    14281 gcggccgctt gccacgattc accggggcgt gcttctgcct cgatgcgttg ccgctgggcg
    14341 gcctgcgcgg ccttcaactt ctccaccagg tcatcaccca gcgccgcgcc gatttgtacc
    14401 gggccggatg gtttgcgacc gctcacgccg attcctcggg cttgggggtt ccagtgccat
    14461 tgcagggccg gcaggcaacc cagccgctta cgcctggcca accgcccgtt cctccacaca
    14521 tggggcattc cacggcgtcg gtgcctggtt gttcttgatt ttccatgccg cctcctttag
    14581 ccgctaaaat tcatctactc atttattcat ttgctcattt actctggtag ctgcgcgatg
    14641 tattcagata gcagctcggt aatggtcttg ccttggcgta ccgcgtacat cttcagcttg
    14701 gtgtgatcct ccgccggcaa ctgaaagttg acccgcttca tggctggcgt gtctgccagg
    14761 ctggccaacg ttgcagcctt gctgctgcgt gcgctcggac ggccggcact tagcgtgttt
    14821 gtgcttttgc tcattttctc tttacctcat taactcaaat gagttttgat ttaatttcag
    14881 cggccagcgc ctggacctcg cgggcagcgt cgccctcggg ttctgattca agaacggttg
    14941 tgccggcggc ggcagtgcct gggtagctca cgcgctgcgt gatacgggac tcaagaatgg
    15001 gcagctcgta cccggccagc gcctcggcaa cctcaccgcc gatgcgcgtg cctttgatcg
    15061 cccgcgacac gacaaaggcc gcttgtagcc ttccatccgt gacctcaatg cgctgcttaa
    15121 ccagctccac caggtcggcg gtggcccata tgtcgtaagg gcttggctgc accggaatca
    15181 gcacgaagtc ggctgccttg atcgcggaca cagccaagtc cgccgcctgg ggcgctccgt
    15241 cgatcactac gaagtcgcgc cggccgatgg ccttcacgtc gcggtcaatc gtcgggcggt
    15301 cgatgccgac aacggttagc ggttgatctt cccgcacggc cgcccaatcg cgggcactgc
    15361 cctggggatc ggaatcgact aacagaacat cggccccggc gagttgcagg gcgcgggcta
    15421 gatgggttgc gatggtcgtc ttgcctgacc cgcctttctg gttaagtaca gcgataacct
    15481 tcatgcgttc cccttgcgta tttgtttatt tactcatcgc atcatatacg cagcgaccgc
    15541 atgacgcaag ctgttttact caaatacaca tcaccttttt agacggcggc gctcggtttc
    15601 ttcagcggcc aagctggccg gccaggccgc cagcttggca tcagacaaac cggccaggat
    15661 ttcatgcagc cgcacggttg agacgtgcgc gggcggctcg aacacgtacc cggccgcgat
    15721 catctccgcc tcgatctctt cggtaatgaa aaacggttcg tcctggccgt cctggtgcgg
    15781 tttcatgctt gttcctcttg gcgttcattc tcggcggccg ccagggcgtc ggcctcggtc
    15841 aatgcgtcct cacggaaggc accgcgccgc ctggcctcgg tgggcgtcac ttcctcgctg
    15901 cgctcaagtg cgcggtacag ggtcgagcga tgcacgccaa gcagtgcagc cgcctctttc
    15961 acggtgcggc cttcctggtc gatcagctcg cgggcgtgcg cgatctgtgc cggggtgagg
    16021 gtagggcggg ggccaaactt cacgcctcgg gccttggcgg cctcgcgccc gctccgggtg
    16081 cggtcgatga ttagggaacg ctcgaactcg gcaatgccgg cgaacacggt caacaccatg
    16141 cggccggccg gcgtggtggt gtcggcccac ggctctgcca ggctacgcag gcccgcgccg
    16201 gcctcctgga tgcgctcggc aatgtccagt aggtcgcggg tgctgcgggc caggcggtct
    16261 agcctggtca ctgtcacaac gtcgccaggg cgtaggtggt caagcatcct ggccagctcc
    16321 gggcggtcgc gcctggtgcc ggtgatcttc tcggaaaaca gcttggtgca gccggccgcg
    16381 tgcagttcgg cccgttggtt ggtcaagtcc tggtcgtcgg tgctgacgcg ggcatagccc
    16441 agcaggccag cggcggcgct cttgttcatg gcgtaatgtc tccggttcta gtcgcaagta
    16501 ttctacttta tgcgactaaa acacgcgaca agaaaacgcc aggaaaaggg cagggcggca
    16561 gcctgtcgcg taacttagga cttgtgcgac atgtcgtttt cagaagacgg ctgcactgaa
    16621 cgtcagaagc cgactgcact atagcagcgg aggggttgga tcaaagtact ttgatcccga
    16681 ggggaaccct gtggttggca tgcacataca aatggacgaa cggataaacc ttttcacgcc
    16741 cttttaaata tccgattatt ctaataaacg ctcttttctc ttaggtttac ccgccaatat
    16801 atcctgtcaa acactgatag tttaaactga aggcgggaaa cgacaatctg atccaagctc
    16861 aagctgctct agcattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
    16921 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
    16981 gtaacgccag ggttttccca gtcacgacgt t
//
