LOCUS       sequence-127777-       16760 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Egg cell-specific promoter-controlled expression of 
            3×FLAG-NLS-zCas9-NLS. Contains gRNA scaffold for insertion of target
            sequence (U6-26 promoter), Hyg resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16760)
  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 16760)
  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: pHEE401
FEATURES             Location/Qualifiers
     source          1..16760
                     /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             3193..3258
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3265..3285
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3310..7410
                     /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             3310..7410
                     /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             7323..7331
                     /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             7411..7458
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(8115..8131)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8115..8131)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8128..8150)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8139..8155
                     /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(8163..8169,8170..8187,8188..8193))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8208..8229
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8420..9097
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     CDS             9165..10190
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     polyA_signal    10230..10404
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    10482..10506
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             10931..11725
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      11812..12400
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12301..12320
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12554..12571
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    12586..12726
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(12652..12671)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     12812..12834
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      13070..13264
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(13330..14403)
                     /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(14832..15461)
                     /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 tggactttga taaatgttcc tcgctgacgt aagaagacat tagtaatggt
     2221 tataatatat agctttctat gaatgtatgg tgagaaaatg tctgttcact gattttgagt
     2281 ttggaataaa agcatttgcg tttggtttat cattgcgttt atacaaggac agagatccac
     2341 tgagctggaa tagcttaaaa ccattatcag aacaaaataa accatttttt gttaagaatc
     2401 agagcatagt aaacaacaga aacaacctaa gagaggtaac ttgtccaaga agatagctaa
     2461 ttatatctat tttataaaag ttatcatagt ttgtaagtca caaaagatgc aaataacaga
     2521 gaaactagga gacttgagaa tatacattct tgtatatttg tattcgagat tgtgaaaatt
     2581 tgaccataag tttaaattct taaaaagata tatctgatct agatgatggt tatagactgt
     2641 aattttacca catgtttaat gatggatagt gacacacatg acacatcgac aacactatag
     2701 catcttattt agattacaac atgaaatttt tctgtaatac atgtctttgt acataattta
     2761 aaagtaattc ctaagaaata tatttataca aggagtttaa agaaaacata gcataaagtt
     2821 caatgagtag taaaaaccat atacagtata tagcataaag ttcaatgagt ttattacaaa
     2881 agcattggtt cactttctgt aacacgacgt taaaccttcg tctccaatag gagcgctact
     2941 gattcaacat gccaatatat actaaatacg tttctacagt caaatgcttt aacgtttcat
     3001 gattaagtga ctatttaccg tcaatccttt cccattcctc ccactaatcc aactttttaa
     3061 ttactcttaa atcaccacta agcttcgaat ccatccaaaa ccacaatata aaaacagaac
     3121 tctcgtaact caatcatcgc aaaacaaaac aaaacaaaac aaaaacccca aaaagaaaga
     3181 ataagctaga tggattacaa ggaccacgac ggggattaca aggaccacga cattgattac
     3241 aaggatgatg atgacaagat ggctccgaag aagaagagga aggttggcat ccacggggtg
     3301 ccagctgctg acaagaagta ctcgatcggc ctcgatattg ggactaactc tgttggctgg
     3361 gccgtgatca ccgacgagta caaggtgccc tcaaagaagt tcaaggtcct gggcaacacc
     3421 gatcggcatt ccatcaagaa gaatctcatt ggcgctctcc tgttcgacag cggcgagacg
     3481 gctgaggcta cgcggctcaa gcgcaccgcc cgcaggcggt acacgcgcag gaagaatcgc
     3541 atctgctacc tgcaggagat tttctccaac gagatggcga aggttgacga ttctttcttc
     3601 cacaggctgg aggagtcatt cctcgtggag gaggataaga agcacgagcg gcatccaatc
     3661 ttcggcaaca ttgtcgacga ggttgcctac cacgagaagt accctacgat ctaccatctg
     3721 cggaagaagc tcgtggactc cacagataag gcggacctcc gcctgatcta cctcgctctg
     3781 gcccacatga ttaagttcag gggccatttc ctgatcgagg gggatctcaa cccggacaat
     3841 agcgatgttg acaagctgtt catccagctc gtgcagacgt acaaccagct cttcgaggag
     3901 aaccccatta atgcgtcagg cgtcgacgcg aaggctatcc tgtccgctag gctctcgaag
     3961 tctcggcgcc tcgagaacct gatcgcccag ctgccgggcg agaagaagaa cggcctgttc
     4021 gggaatctca ttgcgctcag cctggggctc acgcccaact tcaagtcgaa tttcgatctc
     4081 gctgaggacg ccaagctgca gctctccaag gacacatacg acgatgacct ggataacctc
     4141 ctggcccaga tcggcgatca gtacgcggac ctgttcctcg ctgccaagaa tctgtcggac
     4201 gccatcctcc tgtctgatat tctcagggtg aacaccgaga ttacgaaggc tccgctctca
     4261 gcctccatga tcaagcgcta cgacgagcac catcaggatc tgaccctcct gaaggcgctg
     4321 gtcaggcagc agctccccga gaagtacaag gagatcttct tcgatcagtc gaagaacggc
     4381 tacgctgggt acattgacgg cggggcctct caggaggagt tctacaagtt catcaagccg
     4441 attctggaga agatggacgg cacggaggag ctgctggtga agctcaatcg cgaggacctc
     4501 ctgaggaagc agcggacatt cgataacggc agcatcccac accagattca tctcggggag
     4561 ctgcacgcta tcctgaggag gcaggaggac ttctaccctt tcctcaagga taaccgcgag
     4621 aagatcgaga agattctgac tttcaggatc ccgtactacg tcggcccact cgctaggggc
     4681 aactcccgct tcgcttggat gacccgcaag tcagaggaga cgatcacgcc gtggaacttc
     4741 gaggaggtgg tcgacaaggg cgctagcgct cagtcgttca tcgagaggat gacgaatttc
     4801 gacaagaacc tgccaaatga gaaggtgctc cctaagcact cgctcctgta cgagtacttc
     4861 acagtctaca acgagctgac taaggtgaag tatgtgaccg agggcatgag gaagccggct
     4921 ttcctgtctg gggagcagaa gaaggccatc gtggacctcc tgttcaagac caaccggaag
     4981 gtcacggtta agcagctcaa ggaggactac ttcaagaaga ttgagtgctt cgattcggtc
     5041 gagatctctg gcgttgagga ccgcttcaac gcctccctgg ggacctacca cgatctcctg
     5101 aagatcatta aggataagga cttcctggac aacgaggaga atgaggatat cctcgaggac
     5161 attgtgctga cactcactct gttcgaggac cgggagatga tcgaggagcg cctgaagact
     5221 tacgcccatc tcttcgatga caaggtcatg aagcagctca agaggaggag gtacaccggc
     5281 tgggggaggc tgagcaggaa gctcatcaac ggcattcggg acaagcagtc cgggaagacg
     5341 atcctcgact tcctgaagag cgatggcttc gcgaaccgca atttcatgca gctgattcac
     5401 gatgacagcc tcacattcaa ggaggatatc cagaaggctc aggtgagcgg ccagggggac
     5461 tcgctgcacg agcatatcgc gaacctcgct ggctcgccag ctatcaagaa ggggattctg
     5521 cagaccgtga aggttgtgga cgagctggtg aaggtcatgg gcaggcacaa gcctgagaac
     5581 atcgtcattg agatggcccg ggagaatcag accacgcaga agggccagaa gaactcacgc
     5641 gagaggatga agaggatcga ggagggcatt aaggagctgg ggtcccagat cctcaaggag
     5701 cacccggtgg agaacacgca gctgcagaat gagaagctct acctgtacta cctccagaat
     5761 ggccgcgata tgtatgtgga ccaggagctg gatattaaca ggctcagcga ttacgacgtc
     5821 gatcatatcg ttccacagtc attcctgaag gatgactcca ttgacaacaa ggtcctcacc
     5881 aggtcggaca agaaccgggg caagtctgat aatgttcctt cagaggaggt cgttaagaag
     5941 atgaagaact actggcgcca gctcctgaat gccaagctga tcacgcagcg gaagttcgat
     6001 aacctcacaa aggctgagag gggcgggctc tctgagctgg acaaggcggg cttcatcaag
     6061 aggcagctgg tcgagacacg gcagatcact aagcacgttg cgcagattct cgactcacgg
     6121 atgaacacta agtacgatga gaatgacaag ctgatccgcg aggtgaaggt catcaccctg
     6181 aagtcaaagc tcgtctccga cttcaggaag gatttccagt tctacaaggt tcgggagatc
     6241 aacaattacc accatgccca tgacgcgtac ctgaacgcgg tggtcggcac agctctgatc
     6301 aagaagtacc caaagctcga gagcgagttc gtgtacgggg actacaaggt ttacgatgtg
     6361 aggaagatga tcgccaagtc ggagcaggag attggcaagg ctaccgccaa gtacttcttc
     6421 tactctaaca ttatgaattt cttcaagaca gagatcactc tggccaatgg cgagatccgg
     6481 aagcgccccc tcatcgagac gaacggcgag acgggggaga tcgtgtggga caagggcagg
     6541 gatttcgcga ccgtcaggaa ggttctctcc atgccacaag tgaatatcgt caagaagaca
     6601 gaggtccaga ctggcgggtt ctctaaggag tcaattctgc ctaagcggaa cagcgacaag
     6661 ctcatcgccc gcaagaagga ctgggatccg aagaagtacg gcgggttcga cagccccact
     6721 gtggcctact cggtcctggt tgtggcgaag gttgagaagg gcaagtccaa gaagctcaag
     6781 agcgtgaagg agctgctggg gatcacgatt atggagcgct ccagcttcga gaagaacccg
     6841 atcgatttcc tggaggcgaa gggctacaag gaggtgaaga aggacctgat cattaagctc
     6901 cccaagtact cactcttcga gctggagaac ggcaggaagc ggatgctggc ttccgctggc
     6961 gagctgcaga aggggaacga gctggctctg ccgtccaagt atgtgaactt cctctacctg
     7021 gcctcccact acgagaagct caagggcagc cccgaggaca acgagcagaa gcagctgttc
     7081 gtcgagcagc acaagcatta cctcgacgag atcattgagc agatttccga gttctccaag
     7141 cgcgtgatcc tggccgacgc gaatctggat aaggtcctct ccgcgtacaa caagcaccgc
     7201 gacaagccaa tcagggagca ggctgagaat atcattcatc tcttcaccct gacgaacctc
     7261 ggcgcccctg ctgctttcaa gtacttcgac acaactatcg atcgcaagag gtacacaagc
     7321 actaaggagg tcctggacgc gaccctcatc caccagtcga ttaccggcct ctacgagacg
     7381 cgcatcgacc tgtctcagct cgggggcgac aagcggccag cggcgacgaa gaaggcgggg
     7441 caggcgaaga agaagaagtg agctcagagc tttcgttcgt atcatcggtt tcgacaacgt
     7501 tcgtcaagtt caatgcatca gtttcattgc gcacacacca gaatcctact gagtttgagt
     7561 attatggcat tgggaaaact gtttttcttg taccatttgt tgtgcttgta atttactgtg
     7621 ttttttattc ggttttcgct atcgaactgt gaaatggaaa tggatggaga agagttaatg
     7681 aatgatatgg tccttttgtt cattctcaaa ttaatattat ttgttttttc tcttatttgt
     7741 tgtgtgttga atttgaaatt ataagagata tgcaaacatt ttgttttgag taaaaatgtg
     7801 tcaaatcgtg gcctctaatg accgaagtta atatgaggag taaaacactt gtagttgtac
     7861 cattatgctt attcactagg caacaaatat attttcagac ctagaaaagc tgcaaatgtt
     7921 actgaataca agtatgtcct cttgtgtttt agacatttat gaactttcct ttatgtaatt
     7981 ttccagaatc cttgtcagat tctaatcatt gctttataat tatagttata ctcatggatt
     8041 tgtagttgag tatgaaaata ttttttaatg cattttatga cttgccaatt gattgacaac
     8101 gaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     8161 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     8221 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     8281 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg ctagagcagc
     8341 ttgccaacat ggtggagcac gacactctcg tctactccaa gaatatcaaa gatacagtct
     8401 cagaagacca aagggctatt gagacttttc aacaaagggt aatatcggga aacctcctcg
     8461 gattccattg cccagctatc tgtcacttca tcaaaaggac agtagaaaag gaaggtggca
     8521 cctacaaatg ccatcattgc gataaaggaa aggctatcgt tcaagatgcc tctgccgaca
     8581 gtggtcccaa agatggaccc ccacccacga ggagcatcgt ggaaaaagaa gacgttccaa
     8641 ccacgtcttc aaagcaagtg gattgatgtg ataacatggt ggagcacgac actctcgtct
     8701 actccaagaa tatcaaagat acagtctcag aagaccaaag ggctattgag acttttcaac
     8761 aaagggtaat atcgggaaac ctcctcggat tccattgccc agctatctgt cacttcatca
     8821 aaaggacagt agaaaaggaa ggtggcacct acaaatgcca tcattgcgat aaaggaaagg
     8881 ctatcgttca agatgcctct gccgacagtg gtcccaaaga tggaccccca cccacgagga
     8941 gcatcgtgga aaaagaagac gttccaacca cgtcttcaaa gcaagtggat tgatgtgata
     9001 tctccactga cgtaagggat gacgcacaat cccactatcc ttcgcaagac cttcctctat
     9061 ataaggaagt tcatttcatt tggagaggac acgctgaaat caccagtctc tctctacaaa
     9121 tctatctctc tcgagctttc gcagatcccg gggggcaatg agatatgaaa aagcctgaac
     9181 tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga
     9241 tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat
     9301 atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc
     9361 actttgcatc ggccgcgctc ccgattccgg aagtgcttga cattggggag tttagcgaga
     9421 gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa
     9481 ccgaactgcc cgctgttcta caaccggtcg cggaggctat ggatgcgatc gctgcggccg
     9541 atcttagcca gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta
     9601 catggcgtga tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga
     9661 tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg
     9721 aggactgccc cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga
     9781 cggacaatgg ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc
     9841 aatacgaggt cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga
     9901 cgcgctactt cgagcggagg catccggagc ttgcaggatc gccacgactc cgggcgtata
     9961 tgctccgcat tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg
    10021 cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc
    10081 gtacacaaat cgcccgcaga agcgcggccg tctggaccga tggctgtgta gaagtactcg
    10141 ccgatagtgg aaaccgacgc cccagcactc gtccgagggc aaagaaatag agtagatgcc
    10201 gaccggatct gtcgatcgac aagctcgagt ttctccataa taatgtgtga gtagttccca
    10261 gataagggaa ttagggttcc tatagggttt cgctcatgtg ttgagcatat aagaaaccct
    10321 tagtatgtat ttgtatttgt aaaatacttc tatcaataaa atttctaatt cctaaaacca
    10381 aaatccagta ctaaaatcca gatcccccga attaattcgg cgttaattca gtacattaaa
    10441 aacgtccgca atgtgttatt aagttgtcta agcgtcaatt tgtttacacc acaatatatc
    10501 ctgccaccag ccagccaaca gctccccgac cggcagctcg gcacaaaatc accactcgat
    10561 acaggcagcc catcagtccg ggacggcgtc agcgggagag ccgttgtaag gcggcagact
    10621 ttgctcatgt taccgatgct attcggaaga acggcaacta agctgccggg tttgaaacac
    10681 ggatgatctc gcggagggta gcatgttgat tgtaacgatg acagagcgtt gctgcctgtg
    10741 atcaccgcgg tttcaaaatc ggctccgtcg atactatgtt atacgccaac tttgaaaaca
    10801 actttgaaaa agctgttttc tggtatttaa ggttttagaa tgcaaggaac agtgaattgg
    10861 agttcgtctt gttataatta gcttcttggg gtatctttaa atactgtaga aaagaggaag
    10921 gaaataataa atggctaaaa tgagaatatc accggaattg aaaaaactga tcgaaaaata
    10981 ccgctgcgta aaagatacgg aaggaatgtc tcctgctaag gtatataagc tggtgggaga
    11041 aaatgaaaac ctatatttaa aaatgacgga cagccggtat aaagggacca cctatgatgt
    11101 ggaacgggaa aaggacatga tgctatggct ggaaggaaag ctgcctgttc caaaggtcct
    11161 gcactttgaa cggcatgatg gctggagcaa tctgctcatg agtgaggccg atggcgtcct
    11221 ttgctcggaa gagtatgaag atgaacaaag ccctgaaaag attatcgagc tgtatgcgga
    11281 gtgcatcagg ctctttcact ccatcgacat atcggattgt ccctatacga atagcttaga
    11341 cagccgctta gccgaattgg attacttact gaataacgat ctggccgatg tggattgcga
    11401 aaactgggaa gaagacactc catttaaaga tccgcgcgag ctgtatgatt ttttaaagac
    11461 ggaaaagccc gaagaggaac ttgtcttttc ccacggcgac ctgggagaca gcaacatctt
    11521 tgtgaaagat ggcaaagtaa gtggctttat tgatcttggg agaagcggca gggcggacaa
    11581 gtggtatgac attgccttct gcgtccggtc gatcagggag gatatcgggg aagaacagta
    11641 tgtcgagcta ttttttgact tactggggat caagcctgat tgggagaaaa taaaatatta
    11701 tattttactg gatgaattgt tttagtacct agaatgcatg accaaaatcc cttaacgtga
    11761 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
    11821 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
    11881 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
    11941 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
    12001 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
    12061 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
    12121 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
    12181 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
    12241 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
    12301 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
    12361 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
    12421 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
    12481 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
    12541 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
    12601 tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg
    12661 ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg
    12721 gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
    12781 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
    12841 ccgtcatcac cgaaacgcgc gaggcagggt gccttgatgt gggcgccggc ggtcgagtgg
    12901 cgacggcgcg gcttgtccgc gccctggtag attgcctggc cgtaggccag ccatttttga
    12961 gcggccagcg gccgcgatag gccgacgcga agcggcgggg cgtagggagc gcagcgaccg
    13021 aagggtaggc gctttttgca gctcttcggc tgtgcgctgg ccagacagtt atgcacaggc
    13081 caggcgggtt ttaagagttt taataagttt taaagagttt taggcggaaa aatcgccttt
    13141 tttctctttt atatcagtca cttacatgtg tgaccggttc ccaatgtacg gctttgggtt
    13201 cccaatgtac gggttccggt tcccaatgta cggctttggg ttcccaatgt acgtgctatc
    13261 cacaggaaac agaccttttc gacctttttc ccctgctagg gcaatttgcc ctagcatctg
    13321 ctccgtacat taggaaccgg cggatgcttc gccctcgatc aggttgcggt agcgcatgac
    13381 taggatcggg ccagcctgcc ccgcctcctc cttcaaatcg tactccggca ggtcatttga
    13441 cccgatcagc ttgcgcacgg tgaaacagaa cttcttgaac tctccggcgc tgccactgcg
    13501 ttcgtagatc gtcttgaaca accatctggc ttctgccttg cctgcggcgc ggcgtgccag
    13561 gcggtagaga aaacggccga tgccgggatc gatcaaaaag taatcggggt gaaccgtcag
    13621 cacgtccggg ttcttgcctt ctgtgatctc gcggtacatc caatcagcta gctcgatctc
    13681 gatgtactcc ggccgcccgg tttcgctctt tacgatcttg tagcggctaa tcaaggcttc
    13741 accctcggat accgtcacca ggcggccgtt cttggccttc ttcgtacgct gcatggcaac
    13801 gtgcgtggtg tttaaccgaa tgcaggtttc taccaggtcg tctttctgct ttccgccatc
    13861 ggctcgccgg cagaacttga gtacgtccgc aacgtgtgga cggaacacgc ggccgggctt
    13921 gtctcccttc ccttcccggt atcggttcat ggattcggtt agatgggaaa ccgccatcag
    13981 taccaggtcg taatcccaca cactggccat gccggccggc cctgcggaaa cctctacgtg
    14041 cccgtctgga agctcgtagc ggatcacctc gccagctcgt cggtcacgct tcgacagacg
    14101 gaaaacggcc acgtccatga tgctgcgact atcgcgggtg cccacgtcat agagcatcgg
    14161 aacgaaaaaa tctggttgct cgtcgccctt gggcggcttc ctaatcgacg gcgcaccggc
    14221 tgccggcggt tgccgggatt ctttgcggat tcgatcagcg gccgcttgcc acgattcacc
    14281 ggggcgtgct tctgcctcga tgcgttgccg ctgggcggcc tgcgcggcct tcaacttctc
    14341 caccaggtca tcacccagcg ccgcgccgat ttgtaccggg ccggatggtt tgcgaccgct
    14401 cacgccgatt cctcgggctt gggggttcca gtgccattgc agggccggca gacaacccag
    14461 ccgcttacgc ctggccaacc gcccgttcct ccacacatgg ggcattccac ggcgtcggtg
    14521 cctggttgtt cttgattttc catgccgcct cctttagccg ctaaaattca tctactcatt
    14581 tattcatttg ctcatttact ctggtagctg cgcgatgtat tcagatagca gctcggtaat
    14641 ggtcttgcct tggcgtaccg cgtacatctt cagcttggtg tgatcctccg ccggcaactg
    14701 aaagttgacc cgcttcatgg ctggcgtgtc tgccaggctg gccaacgttg cagccttgct
    14761 gctgcgtgcg ctcggacggc cggcacttag cgtgtttgtg cttttgctca ttttctcttt
    14821 acctcattaa ctcaaatgag ttttgattta atttcagcgg ccagcgcctg gacctcgcgg
    14881 gcagcgtcgc cctcgggttc tgattcaaga acggttgtgc cggcggcggc agtgcctggg
    14941 tagctcacgc gctgcgtgat acgggactca agaatgggca gctcgtaccc ggccagcgcc
    15001 tcggcaacct caccgccgat gcgcgtgcct ttgatcgccc gcgacacgac aaaggccgct
    15061 tgtagccttc catccgtgac ctcaatgcgc tgcttaacca gctccaccag gtcggcggtg
    15121 gcccatatgt cgtaagggct tggctgcacc ggaatcagca cgaagtcggc tgccttgatc
    15181 gcggacacag ccaagtccgc cgcctggggc gctccgtcga tcactacgaa gtcgcgccgg
    15241 ccgatggcct tcacgtcgcg gtcaatcgtc gggcggtcga tgccgacaac ggttagcggt
    15301 tgatcttccc gcacggccgc ccaatcgcgg gcactgccct ggggatcgga atcgactaac
    15361 agaacatcgg ccccggcgag ttgcagggcg cgggctagat gggttgcgat ggtcgtcttg
    15421 cctgacccgc ctttctggtt aagtacagcg ataaccttca tgcgttcccc ttgcgtattt
    15481 gtttatttac tcatcgcatc atatacgcag cgaccgcatg acgcaagctg ttttactcaa
    15541 atacacatca cctttttaga cggcggcgct cggtttcttc agcggccaag ctggccggcc
    15601 aggccgccag cttggcatca gacaaaccgg ccaggatttc atgcagccgc acggttgaga
    15661 cgtgcgcggg cggctcgaac acgtacccgg ccgcgatcat ctccgcctcg atctcttcgg
    15721 taatgaaaaa cggttcgtcc tggccgtcct ggtgcggttt catgcttgtt cctcttggcg
    15781 ttcattctcg gcggccgcca gggcgtcggc ctcggtcaat gcgtcctcac ggaaggcacc
    15841 gcgccgcctg gcctcggtgg gcgtcacttc ctcgctgcgc tcaagtgcgc ggtacagggt
    15901 cgagcgatgc acgccaagca gtgcagccgc ctctttcacg gtgcggcctt cctggtcgat
    15961 cagctcgcgg gcgtgcgcga tctgtgccgg ggtgagggta gggcgggggc caaacttcac
    16021 gcctcgggcc ttggcggcct cgcgcccgct ccgggtgcgg tcgatgatta gggaacgctc
    16081 gaactcggca atgccggcga acacggtcaa caccatgcgg ccggccggcg tggtggtgtc
    16141 ggcccacggc tctgccaggc tacgcaggcc cgcgccggcc tcctggatgc gctcggcaat
    16201 gtccagtagg tcgcgggtgc tgcgggccag gcggtctagc ctggtcactg tcacaacgtc
    16261 gccagggcgt aggtggtcaa gcatcctggc cagctccggg cggtcgcgcc tggtgccggt
    16321 gatcttctcg gaaaacagct tggtgcagcc ggccgcgtgc agttcggccc gttggttggt
    16381 caagtcctgg tcgtcggtgc tgacgcgggc atagcccagc aggccagcgg cggcgctctt
    16441 gttcatggcg taatgtctcc ggttctagtc gcaagtattc tactttatgc gactaaaaca
    16501 cgcgacaaga aaacgccagg aaaagggcag ggcggcagcc tgtcgcgtaa cttaggactt
    16561 gtgcgacatg tcgttttcag aagacggctg cactgaacgt cagaagccga ctgcactata
    16621 gcagcggagg ggttggatca aagtactttg atcccgaggg gaaccctgtg gttggcatgc
    16681 acatacaaat ggacgaacgg ataaaccttt tcacgccctt ttaaatatcc gttattctaa
    16741 taaacgctct tttctcttag
//
