LOCUS       sequence-176319-       16625 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR/Cas9-mediated genome editing in Arabidopsis. Contains Cas9 
            and empty gRNA scaffold, Bar resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16625)
  AUTHORS   Wang ZP, Xing HL, Dong L, Zhang HY, Han CY, Wang XC, Chen QJ
  TITLE     Egg cell-specific promoter-controlled CRISPR/Cas9 efficiently 
            generates homozygous mutants for multiple target genes in 
            Arabidopsis in a single generation.
  JOURNAL   Genome Biol. 2015 Jul 21;16:144. doi: 10.1186/s13059-015-0715-0.
  PUBMED    26193878
REFERENCE   2  (bases 1 to 16625)
  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: "Genome 
            Biol."; date: "2015-07-21"; volume: "16:144. doi"; pages: " 
            10.1186/s13059-015-0715-0"
COMMENT     Sequence Label: pXEE401B
FEATURES             Location/Qualifiers
     source          1..16625
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..25
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     213..235
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     227..244
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     228..244
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             957..1748
                     /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        1900..1975
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             3545..3610
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3617..3637
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3662..7762
                     /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             3662..7762
                     /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             7675..7683
                     /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             7763..7810
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     promoter        8567..9243
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     9180..9200
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     misc_feature    10347..10371
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             10796..11590
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      11677..12265
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12166..12185
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12419..12436
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    12451..12591
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(12517..12536)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     12677..12699
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      12935..13129
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(13195..14268)
                     /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(14697..15326)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
ORIGIN
        1 gtttacccgc caatatatcc tgtcaaacac tgatagttta aactgaaggc gggaaacgac
       61 aatctgatcc aagctcaagc tgctctagca ttcgccattc aggctgcgca actgttggga
      121 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
      181 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
      241 cagtgccaag cttcgacttg ccttccgcac aatacatcat ttcttcttag ctttttttct
      301 tcttcttcgt tcatacagtt tttttttgtt tatcagctta cattttcttg aaccgtagct
      361 ttcgttttct tctttttaac tttccattcg gagtttttgt atcttgtttc atagtttgtc
      421 ccaggattag aatgattagg catcgaacct tcaagaattt gattgaataa aacatcttca
      481 ttcttaagat atgaagataa tcttcaaaag gcccctggga atctgaaaga agagaagcag
      541 gcccatttat atgggaaaga acaatagtat ttcttatata ggcccattta agttgaaaac
      601 aatcttcaaa agtcccacat cgcttagata agaaaacgaa gctgagttta tatacagcta
      661 gagtcgaagt agtgattggg agaccaaccc agtggacata agcctgttcg gttcgtaagc
      721 tgtaatgcaa gtagcgtatg cgctcacgca actggtccag aaccttgacc gaacgcagcg
      781 gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt ggggtacagt
      841 ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt ttgatgttat
      901 ggagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta aacatcatgg
      961 gggaagcggt gatcgccgaa gtatcgactc aactatcaga ggtagttggc gtcatcgagc
     1021 gccatctcga accgacgttg ctggccgtac atttgtacgg ctccgcagtg gatggcggcc
     1081 tgaagccaca cagtgatatt gatttgctgg ttacggtgac cgtaaggctt gatgaaacaa
     1141 cgcggcgagc tttgatcaac gaccttttgg aaacttcggc ttcccctgga gagagcgaga
     1201 ttctccgcgc tgtagaagtc accattgttg tgcacgacga catcattccg tggcgttatc
     1261 cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa tgacattctt gcaggtatct
     1321 tcgagccagc cacgatcgac attgatctgg ctatcttgct gacaaaagca agagaacata
     1381 gcgttgcctt ggtaggtcca gcggcggagg aactctttga tccggttcct gaacaggatc
     1441 tatttgaggc gctaaatgaa accttaacgc tatggaactc gccgcccgac tgggctggcg
     1501 atgagcgaaa tgtagtgctt acgttgtccc gcatttggta cagcgcagta accggcaaaa
     1561 tcgcgccgaa ggatgtcgct gccgactggg caatggagcg cctgccggcc cagtatcagc
     1621 ccgtcatact tgaagctaga caggcttatc ttggacaaga agaagatcgc ttggcctcgc
     1681 gcgcagatca gttggaagaa tttgtccact acgtgaaagg cgagatcacc aaggtagtcg
     1741 gcaaataatg tctagctaga aattcgttca agccgacgcc gcttcgcggc gcggcttaac
     1801 tcaagcgtta gatgcactaa gcacataatt gctcacagcc aaactatcag gtcaagtctg
     1861 cttttattat ttttaagcgt gcataataag ccggtctcgg ttttagagct agaaatagca
     1921 agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt
     1981 tttgcaaaat tttccagatc gatttcttct tcctctgttc ttcggcgttc aatttctggg
     2041 gttttctctt cgttttctgt aactgaaacc taaaatttga cctaaaaaaa atctcaaata
     2101 atatgattca gtggttttgt acttttcagt tagttgagtt ttgcagttcc gatgagataa
     2161 accaatacca tggttatact agtgaataaa agcatttgcg tttggtttat cattgcgttt
     2221 atacaaggac agagatccac tgagctggaa tagcttaaaa ccattatcag aacaaaataa
     2281 accatttttt gttaagaatc agagcatagt aaacaacaga aacaacctaa gagaggtaac
     2341 ttgtccaaga agatagctaa ttatatctat tttataaaag ttatcatagt ttgtaagtca
     2401 caaaagatgc aaataacaga gaaactagga gacttgagaa tatacattct tgtatatttg
     2461 tattcgagat tgtgaaaatt tgaccataag tttaaattct taaaaagata tatctgatct
     2521 aggtgatggt tatagactgt aattttacca catgtttaat gatggatagt gacacacatg
     2581 acacatcgac aacactatag catcttattt agattacaac atgaaatttt tctgtaatac
     2641 atgtctttgt acataattta aaagtaattc ctaagaaata tatttataca aggagtttaa
     2701 agaaaacata gcataaagtt caatgagtag taaaaaccat atacagtata tagcataaag
     2761 ttcaatgagt ttattacaaa agcattggtt cactttctgt aacacgacgt taaaccttcg
     2821 tctccaatag gagcgctact gattcaacat gccaatatat actaaatacg tttctacagt
     2881 caaatgcttt aacgtttcat gattaagtga ctatttaccg tcaatccttt cccattcctc
     2941 ccactaatcc aactttttaa ttactcttaa atcaccacta agctagtaac gcctatcatg
     3001 aattagctct actaaatcta gcaacctttc aaatttgcag tattgcaggt gtctctgtgt
     3061 ctttaaaata gttgccttat gatttcttcg gtttcaagat gatcaaatag ttatagattt
     3121 catgctcaca catgctcatt agatgtgtac atactttact tacccaaatc tattttctcg
     3181 caaagatttt gatggtaaag ctgatttggt tctattgaac taaatcaaac gagtttcaga
     3241 ctgagtgatt ctaatccggc ccattagccc ctaaacagac ccactaatta cgcagctttt
     3301 aatagagtaa ttacacctag tttacccact aaaccactaa gcactaatta tctcacaatc
     3361 taatgagctt ccctcgtaat tacttgggct ttcactctac catttatttg taacagtcaa
     3421 gtctctactg tctctatata aactctctaa agttaacaca caattctcat cacaaacaaa
     3481 tcaaccaaag caacttctac tctttcttct ttcgacctta tcaatctgtt gagaaatcta
     3541 gatggattac aaggaccacg acggggatta caaggaccac gacattgatt acaaggatga
     3601 tgatgacaag atggctccga agaagaagag gaaggttggc atccacgggg tgccagctgc
     3661 tgacaagaag tactcgatcg gcctcgatat tgggactaac tctgttggct gggccgtgat
     3721 caccgacgag tacaaggtgc cctcaaagaa gttcaaggtc ctgggcaaca ccgatcggca
     3781 ttccatcaag aagaatctca ttggcgctct cctgttcgac agcggcgaga cggctgaggc
     3841 tacgcggctc aagcgcaccg cccgcaggcg gtacacgcgc aggaagaatc gcatctgcta
     3901 cctgcaggag attttctcca acgagatggc gaaggttgac gattctttct tccacaggct
     3961 ggaggagtca ttcctcgtgg aggaggataa gaagcacgag cggcatccaa tcttcggcaa
     4021 cattgtcgac gaggttgcct accacgagaa gtaccctacg atctaccatc tgcggaagaa
     4081 gctcgtggac tccacagata aggcggacct ccgcctgatc tacctcgctc tggcccacat
     4141 gattaagttc aggggccatt tcctgatcga gggggatctc aacccggaca atagcgatgt
     4201 tgacaagctg ttcatccagc tcgtgcagac gtacaaccag ctcttcgagg agaaccccat
     4261 taatgcgtca ggcgtcgacg cgaaggctat cctgtccgct aggctctcga agtctcggcg
     4321 cctcgagaac ctgatcgccc agctgccggg cgagaagaag aacggcctgt tcgggaatct
     4381 cattgcgctc agcctggggc tcacgcccaa cttcaagtcg aatttcgatc tcgctgagga
     4441 cgccaagctg cagctctcca aggacacata cgacgatgac ctggataacc tcctggccca
     4501 gatcggcgat cagtacgcgg acctgttcct cgctgccaag aatctgtcgg acgccatcct
     4561 cctgtctgat attctcaggg tgaacaccga gattacgaag gctccgctct cagcctccat
     4621 gatcaagcgc tacgacgagc accatcagga tctgaccctc ctgaaggcgc tggtcaggca
     4681 gcagctcccc gagaagtaca aggagatctt cttcgatcag tcgaagaacg gctacgctgg
     4741 gtacattgac ggcggggcct ctcaggagga gttctacaag ttcatcaagc cgattctgga
     4801 gaagatggac ggcacggagg agctgctggt gaagctcaat cgcgaggacc tcctgaggaa
     4861 gcagcggaca ttcgataacg gcagcatccc acaccagatt catctcgggg agctgcacgc
     4921 tatcctgagg aggcaggagg acttctaccc tttcctcaag gataaccgcg agaagatcga
     4981 gaagattctg actttcagga tcccgtacta cgtcggccca ctcgctaggg gcaactcccg
     5041 cttcgcttgg atgacccgca agtcagagga gacgatcacg ccgtggaact tcgaggaggt
     5101 ggtcgacaag ggcgctagcg ctcagtcgtt catcgagagg atgacgaatt tcgacaagaa
     5161 cctgccaaat gagaaggtgc tccctaagca ctcgctcctg tacgagtact tcacagtcta
     5221 caacgagctg actaaggtga agtatgtgac cgagggcatg aggaagccgg ctttcctgtc
     5281 tggggagcag aagaaggcca tcgtggacct cctgttcaag accaaccgga aggtcacggt
     5341 taagcagctc aaggaggact acttcaagaa gattgagtgc ttcgattcgg tcgagatctc
     5401 tggcgttgag gaccgcttca acgcctccct ggggacctac cacgatctcc tgaagatcat
     5461 taaggataag gacttcctgg acaacgagga gaatgaggat atcctcgagg acattgtgct
     5521 gacactcact ctgttcgagg accgggagat gatcgaggag cgcctgaaga cttacgccca
     5581 tctcttcgat gacaaggtca tgaagcagct caagaggagg aggtacaccg gctgggggag
     5641 gctgagcagg aagctcatca acggcattcg ggacaagcag tccgggaaga cgatcctcga
     5701 cttcctgaag agcgatggct tcgcgaaccg caatttcatg cagctgattc acgatgacag
     5761 cctcacattc aaggaggata tccagaaggc tcaggtgagc ggccaggggg actcgctgca
     5821 cgagcatatc gcgaacctcg ctggctcgcc agctatcaag aaggggattc tgcagaccgt
     5881 gaaggttgtg gacgagctgg tgaaggtcat gggcaggcac aagcctgaga acatcgtcat
     5941 tgagatggcc cgggagaatc agaccacgca gaagggccag aagaactcac gcgagaggat
     6001 gaagaggatc gaggagggca ttaaggagct ggggtcccag atcctcaagg agcacccggt
     6061 ggagaacacg cagctgcaga atgagaagct ctacctgtac tacctccaga atggccgcga
     6121 tatgtatgtg gaccaggagc tggatattaa caggctcagc gattacgacg tcgatcatat
     6181 cgttccacag tcattcctga aggatgactc cattgacaac aaggtcctca ccaggtcgga
     6241 caagaaccgg ggcaagtctg ataatgttcc ttcagaggag gtcgttaaga agatgaagaa
     6301 ctactggcgc cagctcctga atgccaagct gatcacgcag cggaagttcg ataacctcac
     6361 aaaggctgag aggggcgggc tctctgagct ggacaaggcg ggcttcatca agaggcagct
     6421 ggtcgagaca cggcagatca ctaagcacgt tgcgcagatt ctcgactcac ggatgaacac
     6481 taagtacgat gagaatgaca agctgatccg cgaggtgaag gtcatcaccc tgaagtcaaa
     6541 gctcgtctcc gacttcagga aggatttcca gttctacaag gttcgggaga tcaacaatta
     6601 ccaccatgcc catgacgcgt acctgaacgc ggtggtcggc acagctctga tcaagaagta
     6661 cccaaagctc gagagcgagt tcgtgtacgg ggactacaag gtttacgatg tgaggaagat
     6721 gatcgccaag tcggagcagg agattggcaa ggctaccgcc aagtacttct tctactctaa
     6781 cattatgaat ttcttcaaga cagagatcac tctggccaat ggcgagatcc ggaagcgccc
     6841 cctcatcgag acgaacggcg agacggggga gatcgtgtgg gacaagggca gggatttcgc
     6901 gaccgtcagg aaggttctct ccatgccaca agtgaatatc gtcaagaaga cagaggtcca
     6961 gactggcggg ttctctaagg agtcaattct gcctaagcgg aacagcgaca agctcatcgc
     7021 ccgcaagaag gactgggatc cgaagaagta cggcgggttc gacagcccca ctgtggccta
     7081 ctcggtcctg gttgtggcga aggttgagaa gggcaagtcc aagaagctca agagcgtgaa
     7141 ggagctgctg gggatcacga ttatggagcg ctccagcttc gagaagaacc cgatcgattt
     7201 cctggaggcg aagggctaca aggaggtgaa gaaggacctg atcattaagc tccccaagta
     7261 ctcactcttc gagctggaga acggcaggaa gcggatgctg gcttccgctg gcgagctgca
     7321 gaaggggaac gagctggctc tgccgtccaa gtatgtgaac ttcctctacc tggcctccca
     7381 ctacgagaag ctcaagggca gccccgagga caacgagcag aagcagctgt tcgtcgagca
     7441 gcacaagcat tacctcgacg agatcattga gcagatttcc gagttctcca agcgcgtgat
     7501 cctggccgac gcgaatctgg ataaggtcct ctccgcgtac aacaagcacc gcgacaagcc
     7561 aatcagggag caggctgaga atatcattca tctcttcacc ctgacgaacc tcggcgcccc
     7621 tgctgctttc aagtacttcg acacaactat cgatcgcaag aggtacacaa gcactaagga
     7681 ggtcctggac gcgaccctca tccaccagtc gattaccggc ctctacgaga cgcgcatcga
     7741 cctgtctcag ctcgggggcg acaagcggcc agcggcgacg aagaaggcgg ggcaggcgaa
     7801 gaagaagaag tgagctcaga gctttcgttc gtatcatcgg tttcgacaac gttcgtcaag
     7861 ttcaatgcat cagtttcatt gcgcacacac cagaatccta ctgagtttga gtattatggc
     7921 attgggaaaa ctgtttttct tgtaccattt gttgtgcttg taatttactg tgttttttat
     7981 tcggttttcg ctatcgaact gtgaaatgga aatggatgga gaagagttaa tgaatgatat
     8041 ggtccttttg ttcattctca aattaatatt atttgttttt tctcttattt gttgtgtgtt
     8101 gaatttgaaa ttataagaga tatgcaaaca ttttgttttg agtaaaaatg tgtcaaatcg
     8161 tggcctctaa tgaccgaagt taatatgagg agtaaaacac ttgtagttgt accattatgc
     8221 ttattcacta ggcaacaaat atattttcag acctagaaaa gctgcaaatg ttactgaata
     8281 caagtatgtc ctcttgtgtt ttagacattt atgaactttc ctttatgtaa ttttccagaa
     8341 tccttgtcag attctaatca ttgctttata attatagtta tactcatgga tttgtagttg
     8401 agtatgaaaa tattttttaa tgcattttat gacttgccaa ttgattgaca acgaattcga
     8461 ggcggtttgc gtattggcta gagcagcttg ccaacatggt ggagcacgac actctcgtct
     8521 actccaagaa tatcaaagat acagtctcag aagaccaaag ggctattgag acttttcaac
     8581 aaagggtaat atcgggaaac ctcctcggat tccattgccc agctatctgt cacttcatca
     8641 aaaggacagt agaaaaggaa ggtggcacct acaaatgcca tcattgcgat aaaggaaagg
     8701 ctatcgttca agatgcctct gccgacagtg gtcccaaaga tggaccccca cccacgagga
     8761 gcatcgtgga aaaagaagac gttccaacca cgtcttcaaa gcaagtggat tgatgtgaac
     8821 atggtggagc acgacactct cgtctactcc aagaatatca aagatacagt ctcagaagac
     8881 caaagggcta ttgagacttt tcaacaaagg gtaatatcgg gaaacctcct cggattccat
     8941 tgcccagcta tctgtcactt catcaaaagg acagtagaaa aggaaggtgg cacctacaaa
     9001 tgccatcatt gcgataaagg aaaggctatc gttcaagatg cctctgccga cagtggtccc
     9061 aaagatggac ccccacccac gaggagcatc gtggaaaaag aagacgttcc aaccacgtct
     9121 tcaaagcaag tggattgatg tgatatctcc actgacgtaa gggatgacgc acaatcccac
     9181 tatccttcgc aagacccttc ctctatataa ggaagttcat ttcatttgga gaggacctcg
     9241 acaccatctt ccacacactc aagccacact attggagaac acacagggac aacacaccat
     9301 aagatccaag ggaggccacc gccgccgcgc cggtaaccac cccgcccctc tcctctttct
     9361 ttctccgttt gatctttctt tcttcttttt gtgggtagaa tttgaatccc tcagcattgt
     9421 tcatcggtag tttttctttt catgatttgt gacaaatgca gcctcgtgcg gagctttttt
     9481 gtaggtagaa gctagatgag ccccgagagg aggccagcgg acattaggag ggcgactgag
     9541 gcggatatgc cagccgtctg caccatcgtc aatcactaca tcgaaacctc cacggtgaac
     9601 ttcaggacag agccgcagga gccccaggag tggactgacg atctcgtccg cctgagggag
     9661 cgctacccgt ggctcgtcgc tgaggttgac ggcgaggtgg ctgggatcgc ttacgctggc
     9721 ccctggaagg cgcgcaatgc ttacgattgg accgccgaga gcacggttta cgtgtcgcca
     9781 aggcaccaga ggaccggcct cgggtcaaca ctgtacactc atctcctgaa gtccctggag
     9841 gctcagggct tcaagagcgt ggtcgccgtt attggcctgc caaacgaccc ttctgtgcgg
     9901 atgcacgagg ccctcggcta cgctccacgc gggatgctga gggctgctgg cttcaagcac
     9961 gggaattggc atgacgtggg cttctggcag ctcgatttct cgctgccggt cccgccgagg
    10021 ccggtgctcc ccgtcacgga gatctagctc gatcgagcgg ccatgctaga gtccgcaaaa
    10081 atcaccagtc tctctctaca aatctatctc tctctatttt tctccagaat aatgtgtgag
    10141 tagttcccag ataagggaat tagggttctt atagggtttc gctcatgtgt tgagcatata
    10201 agaaaccctt agtatgtatt tgtatttgta aaatacttct atcaataaaa tttctaattc
    10261 ctaaaaccaa aatccagtga cctactagta attcagtaca ttaaaaacgt ccgcaatgtg
    10321 ttattaagtt gtctaagcgt caatttgttt acaccacaat atatcctgcc accagccagc
    10381 caacagctcc ccgaccggca gctcggcaca aaatcaccac tcgatacagg cagcccatca
    10441 gtccgggacg gcgtcagcgg gagagccgtt gtaaggcggc agactttgct catgttaccg
    10501 atgctattcg gaagaacggc aactaagctg ccgggtttga aacacggatg atctcgcgga
    10561 gggtagcatg ttgattgtaa cgatgacaga gcgttgctgc ctgtgatcac cgcggtttca
    10621 aaatcggctc cgtcgatact atgttatacg ccaactttga aaacaacttt gaaaaagctg
    10681 ttttctggta tttaaggttt tagaatgcaa ggaacagtga attggagttc gtcttgttat
    10741 aattagcttc ttggggtatc tttaaatact gtagaaaaga ggaaggaaat aataaatggc
    10801 taaaatgaga atatcaccgg aattgaaaaa actgatcgaa aaataccgct gcgtaaaaga
    10861 tacggaagga atgtctcctg ctaaggtata taagctggtg ggagaaaatg aaaacctata
    10921 tttaaaaatg acggacagcc ggtataaagg gaccacctat gatgtggaac gggaaaagga
    10981 catgatgcta tggctggaag gaaagctgcc tgttccaaag gtcctgcact ttgaacggca
    11041 tgatggctgg agcaatctgc tcatgagtga ggccgatggc gtcctttgct cggaagagta
    11101 tgaagatgaa caaagccctg aaaagattat cgagctgtat gcggagtgca tcaggctctt
    11161 tcactccatc gacatatcgg attgtcccta tacgaatagc ttagacagcc gcttagccga
    11221 attggattac ttactgaata acgatctggc cgatgtggat tgcgaaaact gggaagaaga
    11281 cactccattt aaagatccgc gcgagctgta tgatttttta aagacggaaa agcccgaaga
    11341 ggaacttgtc ttttcccacg gcgacctggg agacagcaac atctttgtga aagatggcaa
    11401 agtaagtggc tttattgatc ttgggagaag cggcagggcg gacaagtggt atgacattgc
    11461 cttctgcgtc cggtcgatca gggaggatat cggggaagaa cagtatgtcg agctattttt
    11521 tgacttactg gggatcaagc ctgattggga gaaaataaaa tattatattt tactggatga
    11581 attgttttag tacctagaat gcatgaccaa aatcccttaa cgtgagtttt cgttccactg
    11641 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
    11701 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
    11761 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
    11821 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
    11881 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
    11941 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
    12001 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
    12061 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
    12121 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
    12181 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
    12241 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
    12301 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
    12361 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
    12421 cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct
    12481 gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata
    12541 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
    12601 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
    12661 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
    12721 cgcgcgaggc agggtgcctt gatgtgggcg ccggcggtcg agtggcgacg gcgcggcttg
    12781 tccgcgccct ggtagattgc ctggccgtag gccagccatt tttgagcggc cagcggccgc
    12841 gataggccga cgcgaagcgg cggggcgtag ggagcgcagc gaccgaaggg taggcgcttt
    12901 ttgcagctct tcggctgtgc gctggccaga cagttatgca caggccaggc gggttttaag
    12961 agttttaata agttttaaag agttttaggc ggaaaaatcg ccttttttct cttttatatc
    13021 agtcacttac atgtgtgacc ggttcccaat gtacggcttt gggttcccaa tgtacgggtt
    13081 ccggttccca atgtacggct ttgggttccc aatgtacgtg ctatccacag gaaacagacc
    13141 ttttcgacct ttttcccctg ctagggcaat ttgccctagc atctgctccg tacattagga
    13201 accggcggat gcttcgccct cgatcaggtt gcggtagcgc atgactagga tcgggccagc
    13261 ctgccccgcc tcctccttca aatcgtactc cggcaggtca tttgacccga tcagcttgcg
    13321 cacggtgaaa cagaacttct tgaactctcc ggcgctgcca ctgcgttcgt agatcgtctt
    13381 gaacaaccat ctggcttctg ccttgcctgc ggcgcggcgt gccaggcggt agagaaaacg
    13441 gccgatgccg ggatcgatca aaaagtaatc ggggtgaacc gtcagcacgt ccgggttctt
    13501 gccttctgtg atctcgcggt acatccaatc agctagctcg atctcgatgt actccggccg
    13561 cccggtttcg ctctttacga tcttgtagcg gctaatcaag gcttcaccct cggataccgt
    13621 caccaggcgg ccgttcttgg ccttcttcgt acgctgcatg gcaacgtgcg tggtgtttaa
    13681 ccgaatgcag gtttctacca ggtcgtcttt ctgctttccg ccatcggctc gccggcagaa
    13741 cttgagtacg tccgcaacgt gtggacggaa cacgcggccg ggcttgtctc ccttcccttc
    13801 ccggtatcgg ttcatggatt cggttagatg ggaaaccgcc atcagtacca ggtcgtaatc
    13861 ccacacactg gccatgccgg ccggccctgc ggaaacctct acgtgcccgt ctggaagctc
    13921 gtagcggatc acctcgccag ctcgtcggtc acgcttcgac agacggaaaa cggccacgtc
    13981 catgatgctg cgactatcgc gggtgcccac gtcatagagc atcggaacga aaaaatctgg
    14041 ttgctcgtcg cccttgggcg gcttcctaat cgacggcgca ccggctgccg gcggttgccg
    14101 ggattctttg cggattcgat cagcggccgc ttgccacgat tcaccggggc gtgcttctgc
    14161 ctcgatgcgt tgccgctggg cggcctgcgc ggccttcaac ttctccacca ggtcatcacc
    14221 cagcgccgcg ccgatttgta ccgggccgga tggtttgcga ccgctcacgc cgattcctcg
    14281 ggcttggggg ttccagtgcc attgcagggc cggcagacaa cccagccgct tacgcctggc
    14341 caaccgcccg ttcctccaca catggggcat tccacggcgt cggtgcctgg ttgttcttga
    14401 ttttccatgc cgcctccttt agccgctaaa attcatctac tcatttattc atttgctcat
    14461 ttactctggt agctgcgcga tgtattcaga tagcagctcg gtaatggtct tgccttggcg
    14521 taccgcgtac atcttcagct tggtgtgatc ctccgccggc aactgaaagt tgacccgctt
    14581 catggctggc gtgtctgcca ggctggccaa cgttgcagcc ttgctgctgc gtgcgctcgg
    14641 acggccggca cttagcgtgt ttgtgctttt gctcattttc tctttacctc attaactcaa
    14701 atgagttttg atttaatttc agcggccagc gcctggacct cgcgggcagc gtcgccctcg
    14761 ggttctgatt caagaacggt tgtgccggcg gcggcagtgc ctgggtagct cacgcgctgc
    14821 gtgatacggg actcaagaat gggcagctcg tacccggcca gcgcctcggc aacctcaccg
    14881 ccgatgcgcg tgcctttgat cgcccgcgac acgacaaagg ccgcttgtag ccttccatcc
    14941 gtgacctcaa tgcgctgctt aaccagctcc accaggtcgg cggtggccca tatgtcgtaa
    15001 gggcttggct gcaccggaat cagcacgaag tcggctgcct tgatcgcgga cacagccaag
    15061 tccgccgcct ggggcgctcc gtcgatcact acgaagtcgc gccggccgat ggccttcacg
    15121 tcgcggtcaa tcgtcgggcg gtcgatgccg acaacggtta gcggttgatc ttcccgcacg
    15181 gccgcccaat cgcgggcact gccctgggga tcggaatcga ctaacagaac atcggccccg
    15241 gcgagttgca gggcgcgggc tagatgggtt gcgatggtcg tcttgcctga cccgcctttc
    15301 tggttaagta cagcgataac cttcatgcgt tccccttgcg tatttgttta tttactcatc
    15361 gcatcatata cgcagcgacc gcatgacgca agctgtttta ctcaaataca catcaccttt
    15421 ttagacggcg gcgctcggtt tcttcagcgg ccaagctggc cggccaggcc gccagcttgg
    15481 catcagacaa accggccagg atttcatgca gccgcacggt tgagacgtgc gcgggcggct
    15541 cgaacacgta cccggccgcg atcatctccg cctcgatctc ttcggtaatg aaaaacggtt
    15601 cgtcctggcc gtcctggtgc ggtttcatgc ttgttcctct tggcgttcat tctcggcggc
    15661 cgccagggcg tcggcctcgg tcaatgcgtc ctcacggaag gcaccgcgcc gcctggcctc
    15721 ggtgggcgtc acttcctcgc tgcgctcaag tgcgcggtac agggtcgagc gatgcacgcc
    15781 aagcagtgca gccgcctctt tcacggtgcg gccttcctgg tcgatcagct cgcgggcgtg
    15841 cgcgatctgt gccggggtga gggtagggcg ggggccaaac ttcacgcctc gggccttggc
    15901 ggcctcgcgc ccgctccggg tgcggtcgat gattagggaa cgctcgaact cggcaatgcc
    15961 ggcgaacacg gtcaacacca tgcggccggc cggcgtggtg gtgtcggccc acggctctgc
    16021 caggctacgc aggcccgcgc cggcctcctg gatgcgctcg gcaatgtcca gtaggtcgcg
    16081 ggtgctgcgg gccaggcggt ctagcctggt cactgtcaca acgtcgccag ggcgtaggtg
    16141 gtcaagcatc ctggccagct ccgggcggtc gcgcctggtg ccggtgatct tctcggaaaa
    16201 cagcttggtg cagccggccg cgtgcagttc ggcccgttgg ttggtcaagt cctggtcgtc
    16261 ggtgctgacg cgggcatagc ccagcaggcc agcggcggcg ctcttgttca tggcgtaatg
    16321 tctccggttc tagtcgcaag tattctactt tatgcgacta aaacacgcga caagaaaacg
    16381 ccaggaaaag ggcagggcgg cagcctgtcg cgtaacttag gacttgtgcg acatgtcgtt
    16441 ttcagaagac ggctgcactg aacgtcagaa gccgactgca ctatagcagc ggaggggttg
    16501 gatcaaagta ctttgatccc gaggggaacc ctgtggttgg catgcacata caaatggacg
    16561 aacggataaa ccttttcacg cccttttaaa tatccgttat tctaataaac gctcttttct
    16621 cttag
//
