LOCUS       sequence-452219-       16029 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T-DNA vector for SpCas9 mediated mutagenesis for dicot plants; NGG 
            PAM; Cas9 was driven by 2x35s and the sgRNA was driven by AtU3 
            promoter; Kanamycin for plants selection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16029)
  AUTHORS   Cheng Y, Li G, Qi A, Mandlik R, Pan C, Wang D, Ge S, Qi Y
  TITLE     A comprehensive all-in-one CRISPR toolbox for large-scale screens in
            plants.
  JOURNAL   Plant Cell. 2025 Apr 9:koaf081. doi: 10.1093/plcell/koaf081.
  PUBMED    40202185
REFERENCE   2  (bases 1 to 16029)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant Cell.
            2025 Apr 9:koaf081. doi: 10.1093/plcell/koaf081."
COMMENT     Sequence Label: pYPQAT03
FEATURES             Location/Qualifiers
     source          1..16029
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             425..1219
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      1306..1894
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1795..1814
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2048..2065
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2080..2220
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2146..2165)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2306..2328
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      2564..2758
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(2824..3897)
                     /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(4326..4955)
                     /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    6256..6280
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     6483..6499
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6926..7270
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     protein_bind    7330..7354
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             7376..7441
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             7448..7468
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7493..11593
                     /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             7493..11593
                     /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             11506..11514
                     /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             11594..11641
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     protein_bind    12316..12336
                     /gene="mutant version of attB"
                     /label=attB5
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     CDS             complement(12697..13002)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGAGL"
     primer_bind     12867..12883
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12893..12912
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        12893..12911
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(12959..12979)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(12959..12977)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(12998..13014)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    13022..13038
                     /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(13046..13052,13053..13070,13071..13076))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13091..13112
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_RNA        13154..13229
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(13432..13456)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    13790..13846
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(13855..13871)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    13879..13895
                     /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(13903..13909,13910..13927,13928..13933))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13948..13969
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        14160..14836
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     CDS             14899..15696
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLL
                     SSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEH
                     QGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDI
                     ALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(14956..14975)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     15566..15585
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    15753..15927
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    16005..16029
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 ccagccagcc aacagctccc cgaccggcag ctcggcacaa aatcaccact cgatacaggc
       61 agcccatcag tccgggacgg cgtcagcggg agagccgttg taaggcggca gactttgctc
      121 atgttaccga tgctattcgg aagaacggca actaagctgc cgggtttgaa acacggatga
      181 tctcgcggag ggtagcatgt tgattgtaac gatgacagag cgttgctgcc tgtgatcacc
      241 gcggtttcaa aatcggctcc gtcgatacta tgttatacgc caactttgaa aacaactttg
      301 aaaaagctgt tttctggtat ttaaggtttt agaatgcaag gaacagtgaa ttggagttcg
      361 tcttgttata attagcttct tggggtatct ttaaatactg tagaaaagag gaaggaaata
      421 ataaatggct aaaatgagaa tatcaccgga attgaaaaaa ctgatcgaaa aataccgctg
      481 cgtaaaagat acggaaggaa tgtctcctgc taaggtatat aagctggtgg gagaaaatga
      541 aaacctatat ttaaaaatga cggacagccg gtataaaggg accacctatg atgtggaacg
      601 ggaaaaggac atgatgctat ggctggaagg aaagctgcct gttccaaagg tcctgcactt
      661 tgaacggcat gatggctgga gcaatctgct catgagtgag gccgatggcg tcctttgctc
      721 ggaagagtat gaagatgaac aaagccctga aaagattatc gagctgtatg cggagtgcat
      781 caggctcttt cactccatcg acatatcgga ttgtccctat acgaatagct tagacagccg
      841 cttagccgaa ttggattact tactgaataa cgatctggcc gatgtggatt gcgaaaactg
      901 ggaagaagac actccattta aagatccgcg cgagctgtat gattttttaa agacggaaaa
      961 gcccgaagag gaacttgtct tttcccacgg cgacctggga gacagcaaca tctttgtgaa
     1021 agatggcaaa gtaagtggct ttattgatct tgggagaagc ggcagggcgg acaagtggta
     1081 tgacattgcc ttctgcgtcc ggtcgatcag ggaggatatc ggggaagaac agtatgtcga
     1141 gctatttttt gacttactgg ggatcaagcc tgattgggag aaaataaaat attatatttt
     1201 actggatgaa ttgttttagt acctagaatg catgaccaaa atcccttaac gtgagttttc
     1261 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     1321 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     1381 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     1441 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     1501 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     1561 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     1621 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     1681 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     1741 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     1801 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     1861 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     1921 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     1981 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     2041 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct
     2101 tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga
     2161 tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg
     2221 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     2281 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     2341 tcaccgaaac gcgcgaggca gggtgccttg atgtgggcgc cggcggtcga gtggcgacgg
     2401 cgcggcttgt ccgcgccctg gtagattgcc tggccgtagg ccagccattt ttgagcggcc
     2461 agcggccgcg ataggccgac gcgaagcggc ggggcgtagg gagcgcagcg accgaagggt
     2521 aggcgctttt tgcagctctt cggctgtgcg ctggccagac agttatgcac aggccaggcg
     2581 ggttttaaga gttttaataa gttttaaaga gttttaggcg gaaaaatcgc cttttttctc
     2641 ttttatatca gtcacttaca tgtgtgaccg gttcccaatg tacggctttg ggttcccaat
     2701 gtacgggttc cggttcccaa tgtacggctt tgggttccca atgtacgtgc tatccacagg
     2761 aaacagacct tttcgacctt tttcccctgc tagggcaatt tgccctagca tctgctccgt
     2821 acattaggaa ccggcggatg cttcgccctc gatcaggttg cggtagcgca tgactaggat
     2881 cgggccagcc tgccccgcct cctccttcaa atcgtactcc ggcaggtcat ttgacccgat
     2941 cagcttgcgc acggtgaaac agaacttctt gaactctccg gcgctgccac tgcgttcgta
     3001 gatcgtcttg aacaaccatc tggcttctgc cttgcctgcg gcgcggcgtg ccaggcggta
     3061 gagaaaacgg ccgatgccgg gatcgatcaa aaagtaatcg gggtgaaccg tcagcacgtc
     3121 cgggttcttg ccttctgtga tctcgcggta catccaatca gctagctcga tctcgatgta
     3181 ctccggccgc ccggtttcgc tctttacgat cttgtagcgg ctaatcaagg cttcaccctc
     3241 ggataccgtc accaggcggc cgttcttggc cttcttcgta cgctgcatgg caacgtgcgt
     3301 ggtgtttaac cgaatgcagg tttctaccaa gtcgtctttc tgctttccgc catcggctcg
     3361 ccggcagaac ttgagtacgt ccgcaacgtg tggacggaac acgcggccgg gcttgtctcc
     3421 cttcccttcc cggtatcggt tcatggattc ggttagatgg gaaaccgcca tcagtaccag
     3481 gtcgtaatcc cacacactgg ccatgccggc cggccctgcg gaaacctcta cgtgcccgtc
     3541 tggaagctcg tagcggatca cctcgccagc tcgtcggtca cgcttcgaca gacggaaaac
     3601 ggccacgtcc atgatgctgc gactatcgcg ggtgcccacg tcatagagca tcggaacgaa
     3661 aaaatctggt tgctcgtcgc ccttgggcgg cttcctaatc gacggcgcac cggctgccgg
     3721 cggttgccgg gattctttgc ggattcgatc agcggccgct tgccacgatt caccggggcg
     3781 tgcttctgcc tcgatgcgtt gccgctgggc ggcctgcgcg gccttcaact tctccaccag
     3841 gtcatcaccc agcgccgcgc cgatttgtac cgggccggat ggtttgcgac cgctcacgcc
     3901 gattcctcgg gcttgggggt tccagtgcca ttgcagggcc ggcaggcaac ccagccgctt
     3961 acgcctggcc aaccgcccgt tcctccacac atggggcatt ccacggcgtc ggtgcctggt
     4021 tgttcttgat tttccatgcc gcctccttta gccgctaaaa ttcatctact catttattca
     4081 tttgctcatt tactctggta gctgcgcgat gtattcagat agcagctcgg taatggtctt
     4141 gccttggcgt accgcgtaca tcttcagctt ggtgtgatcc tccgccggca actgaaagtt
     4201 gacccgcttc atggctggcg tgtctgccag gctggccaac gttgcagcct tgctgctgcg
     4261 tgcgctcgga cggccggcac ttagcgtgtt tgtgcttttg ctcattttct ctttacctca
     4321 ttaactcaaa tgagttttga tttaatttca gcggccagcg cctggacctc gcgggcagcg
     4381 tcgccctcgg gttctgattc aagaacggtt gtgccggcgg cggcagtgcc tgggtagctc
     4441 acgcgctgcg tgatacggga ctcaagaatg ggcagctcgt acccggccag cgcctcggca
     4501 acctcaccgc cgatgcgcgt gcctttgatc gcccgcgaca cgacaaaggc cgcttgtagc
     4561 cttccatccg tgacctcaat gcgctgctta accagctcca ccaggtcggc ggtggcccat
     4621 atgtcgtaag ggcttggctg caccggaatc agcacgaagt cggctgcctt gatcgcggac
     4681 acagccaagt ccgccgcctg gggcgctccg tcgatcacta cgaagtcgcg ccggccgatg
     4741 gccttcacgt cgcggtcaat cgtcgggcgg tcgatgccga caacggttag cggttgatct
     4801 tcccgcacgg ccgcccaatc gcgggcactg ccctggggat cggaatcgac taacagaaca
     4861 tcggccccgg cgagttgcag ggcgcgggct agatgggttg cgatggtcgt cttgcctgac
     4921 ccgcctttct ggttaagtac agcgataacc ttcatgcgtt ccccttgcgt atttgtttat
     4981 ttactcatcg catcatatac gcagcgaccg catgacgcaa gctgttttac tcaaatacac
     5041 atcacctttt tagacggcgg cgctcggttt cttcagcggc caagctggcc ggccaggccg
     5101 ccagcttggc atcagacaaa ccggccagga tttcatgcag ccgcacggtt gagacgtgcg
     5161 cgggcggctc gaacacgtac ccggccgcga tcatctccgc ctcgatctct tcggtaatga
     5221 aaaacggttc gtcctggccg tcctggtgcg gtttcatgct tgttcctctt ggcgttcatt
     5281 ctcggcggcc gccagggcgt cggcctcggt caatgcgtcc tcacggaagg caccgcgccg
     5341 cctggcctcg gtgggcgtca cttcctcgct gcgctcaagt gcgcggtaca gggtcgagcg
     5401 atgcacgcca agcagtgcag ccgcctcttt cacggtgcgg ccttcctggt cgatcagctc
     5461 gcgggcgtgc gcgatctgtg ccggggtgag ggtagggcgg gggccaaact tcacgcctcg
     5521 ggccttggcg gcctcgcgcc cgctccgggt gcggtcgatg attagggaac gctcgaactc
     5581 ggcaatgccg gcgaacacgg tcaacaccat gcggccggcc ggcgtggtgg tgtcggccca
     5641 cggctctgcc aggctacgca ggcccgcgcc ggcctcctgg atgcgctcgg caatgtccag
     5701 taggtcgcgg gtgctgcggg ccaggcggtc tagcctggtc actgtcacaa cgtcgccagg
     5761 gcgtaggtgg tcaagcatcc tggccagctc cgggcggtcg cgcctggtgc cggtgatctt
     5821 ctcggaaaac agcttggtgc agccggccgc gtgcagttcg gcccgttggt tggtcaagtc
     5881 ctggtcgtcg gtgctgacgc gggcatagcc cagcaggcca gcggcggcgc tcttgttcat
     5941 ggcgtaatgt ctccggttct agtcgcaagt attctacttt atgcgactaa aacacgcgac
     6001 aagaaaacgc caggaaaagg gcagggcggc agcctgtcgc gtaacttagg acttgtgcga
     6061 catgtcgttt tcagaagacg gctgcactga acgtcagaag ccgactgcac tatagcagcg
     6121 gaggggttgg atcaaagtac tttgatcccg aggggaaccc tgtggttggc atgcacatac
     6181 aaatggacga acggataaac cttttcacgc ccttttaaat atccgattat tctaataaac
     6241 gctcttttct cttaggttta cccgccaata tatcctgtca aacactgata gtttaaactg
     6301 aaggcgggaa acgacaatct gatccaagct caagctgctc tagcattcgc cattcaggct
     6361 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     6421 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     6481 ttgtaaaacg acggccagtg ccaagcttgg cgtgcctgca ggtcaacatg gtggagcacg
     6541 acacacttgt ctactccaaa aatatcaaag atacagtctc agaagaccaa agggcaattg
     6601 agacttttca acaaagggta atatccggaa acctcctcgg attccattgc ccagctatct
     6661 gtcactttat tgtgaagata gtggaaaagg aaggtggctc ctacaaatgc catcattgcg
     6721 ataaaggaaa ggccatcgtt gaagatgcct ctgccgacag tggtcccaaa gatggacccc
     6781 cacccacgag gagcatcgtg gaaaaagaag acgttccaac cacgtcttca aagcaagtgg
     6841 attgatgtga taacatggtg gagcacgaca cacttgtcta ctccaaaaat atcaaagata
     6901 cagtctcaga agaccaaagg gcaattgaga cttttcaaca aagggtaata tccggaaacc
     6961 tcctcggatt ccattgccca gctatctgtc actttattgt gaagatagtg gaaaaggaag
     7021 gtggctccta caaatgccat cattgcgata aaggaaaggc catcgttgaa gatgcctctg
     7081 ccgacagtgg tcccaaagat ggacccccac ccacgaggag catcgtggaa aaagaagacg
     7141 ttccaaccac gtcttcaaag caagtggatt gatgtgatat ctccactgac gtaagggatg
     7201 acgcacaatc ccactatcct tcgcaagacc cttcctctat ataaggaagt tcatttcatt
     7261 tggagaggac ctcgactcta gaggatcccc gggtaccggg ccccccctcg aggcgcgcca
     7321 agctatcaaa caagtttgta caaaaaagca ggctccgaat tcgcccttca ccatggatta
     7381 caaggaccac gacggggatt acaaggacca cgacattgat tacaaggatg atgatgacaa
     7441 gatggctccg aagaagaaga ggaaggttgg catccacggg gtgccagctg ctgacaagaa
     7501 gtactcgatc ggcctcgata ttgggactaa ctctgttggc tgggccgtga tcaccgacga
     7561 gtacaaggtg ccctcaaaga agttcaaggt cctgggcaac accgatcggc attccatcaa
     7621 gaagaatctc attggcgctc tcctgttcga cagcggcgag acggctgagg ctacgcggct
     7681 caagcgcacc gcccgcaggc ggtacacgcg caggaagaat cgcatctgct acctgcagga
     7741 gattttctcc aacgagatgg cgaaggttga cgattctttc ttccacaggc tggaggagtc
     7801 attcctcgtg gaggaggata agaagcacga gcggcatcca atcttcggca acattgtcga
     7861 cgaggttgcc taccacgaga agtaccctac gatctaccat ctgcggaaga agctcgtgga
     7921 ctccacagat aaggcggacc tccgcctgat ctacctcgct ctggcccaca tgattaagtt
     7981 caggggccat ttcctgatcg agggggatct caacccggac aatagcgatg ttgacaagct
     8041 gttcatccag ctcgtgcaga cgtacaacca gctcttcgag gagaacccca ttaatgcgtc
     8101 aggcgtcgac gcgaaggcta tcctgtccgc taggctctcg aagtctcggc gcctcgagaa
     8161 cctgatcgcc cagctgccgg gcgagaagaa gaacggcctg ttcgggaatc tcattgcgct
     8221 cagcctgggg ctcacgccca acttcaagtc gaatttcgat ctcgctgagg acgccaagct
     8281 gcagctctcc aaggacacat acgacgatga cctggataac ctcctggccc agatcggcga
     8341 tcagtacgcg gacctgttcc tcgctgccaa gaatctgtcg gacgccatcc tcctgtctga
     8401 tattctcagg gtgaacaccg agattacgaa ggctccgctc tcagcctcca tgatcaagcg
     8461 ctacgacgag caccatcagg atctgaccct cctgaaggcg ctggtcaggc agcagctccc
     8521 cgagaagtac aaggagatct tcttcgatca gtcgaagaac ggctacgctg ggtacattga
     8581 cggcggggcc tctcaggagg agttctacaa gttcatcaag ccgattctgg agaagatgga
     8641 cggcacggag gagctgctgg tgaagctcaa tcgcgaggac ctcctgagga agcagcggac
     8701 attcgataac ggcagcatcc cacaccagat tcatctcggg gagctgcacg ctatcctgag
     8761 gaggcaggag gacttctacc ctttcctcaa ggataaccgc gagaagatcg agaagattct
     8821 gactttcagg atcccgtact acgtcggccc actcgctagg ggcaactccc gcttcgcttg
     8881 gatgacccgc aagtcagagg agacgatcac gccgtggaac ttcgaggagg tggtcgacaa
     8941 gggcgctagc gctcagtcgt tcatcgagag gatgacgaat ttcgacaaga acctgccaaa
     9001 tgagaaggtg ctccctaagc actcgctcct gtacgagtac ttcacagtct acaacgagct
     9061 gactaaggtg aagtatgtga ccgagggcat gaggaagccg gctttcctgt ctggggagca
     9121 gaagaaggcc atcgtggacc tcctgttcaa gaccaaccgg aaggtcacgg ttaagcagct
     9181 caaggaggac tacttcaaga agattgagtg cttcgattcg gtcgagatct ctggcgttga
     9241 ggaccgcttc aacgcctccc tggggaccta ccacgatctc ctgaagatca ttaaggataa
     9301 ggacttcctg gacaacgagg agaatgagga tatcctcgag gacattgtgc tgacactcac
     9361 tctgttcgag gaccgggaga tgatcgagga gcgcctgaag acttacgccc atctcttcga
     9421 tgacaaggtc atgaagcagc tcaagaggag gaggtacacc ggctggggga ggctgagcag
     9481 gaagctcatc aacggcattc gggacaagca gtccgggaag acgatcctcg acttcctgaa
     9541 gagcgatggc ttcgcgaacc gcaatttcat gcagctgatt cacgatgaca gcctcacatt
     9601 caaggaggat atccagaagg ctcaggtgag cggccagggg gactcgctgc acgagcatat
     9661 cgcgaacctc gctggctcgc cagctatcaa gaaggggatt ctgcagaccg tgaaggttgt
     9721 ggacgagctg gtgaaggtca tgggcaggca caagcctgag aacatcgtca ttgagatggc
     9781 ccgggagaat cagaccacgc agaagggcca gaagaactca cgcgagagga tgaagaggat
     9841 cgaggagggc attaaggagc tggggtccca gatcctcaag gagcacccgg tggagaacac
     9901 gcagctgcag aatgagaagc tctacctgta ctacctccag aatggccgcg atatgtatgt
     9961 ggaccaggag ctggatatta acaggctcag cgattacgac gtcgatcata tcgttccaca
    10021 gtcattcctg aaggatgact ccattgacaa caaggtcctc accaggtcgg acaagaaccg
    10081 gggcaagtct gataatgttc cttcagagga ggtcgttaag aagatgaaga actactggcg
    10141 ccagctcctg aatgccaagc tgatcacgca gcggaagttc gataacctca caaaggctga
    10201 gaggggcggg ctctctgagc tggacaaggc gggcttcatc aagaggcagc tggtcgagac
    10261 acggcagatc actaagcacg ttgcgcagat tctcgactca cggatgaaca ctaagtacga
    10321 tgagaatgac aagctgatcc gcgaggtgaa ggtcatcacc ctgaagtcaa agctcgtctc
    10381 cgacttcagg aaggatttcc agttctacaa ggttcgggag atcaacaatt accaccatgc
    10441 ccatgacgcg tacctgaacg cggtggtcgg cacagctctg atcaagaagt acccaaagct
    10501 cgagagcgag ttcgtgtacg gggactacaa ggtttacgat gtgaggaaga tgatcgccaa
    10561 gtcggagcag gagattggca aggctaccgc caagtacttc ttctactcta acattatgaa
    10621 tttcttcaag acagagatca ctctggccaa tggcgagatc cggaagcgcc ccctcatcga
    10681 gacgaacggc gagacggggg agatcgtgtg ggacaagggc agggatttcg cgaccgtcag
    10741 gaaggttctc tccatgccac aagtgaatat cgtcaagaag acagaggtcc agactggcgg
    10801 gttctctaag gagtcaattc tgcctaagcg gaacagcgac aagctcatcg cccgcaagaa
    10861 ggactgggat ccgaagaagt acggcgggtt cgacagcccc actgtggcct actcggtcct
    10921 ggttgtggcg aaggttgaga agggcaagtc caagaagctc aagagcgtga aggagctgct
    10981 ggggatcacg attatggagc gctccagctt cgagaagaac ccgatcgatt tcctggaggc
    11041 gaagggctac aaggaggtga agaaggacct gatcattaag ctccccaagt actcactctt
    11101 cgagctggag aacggcagga agcggatgct ggcttccgct ggcgagctgc agaaggggaa
    11161 cgagctggct ctgccgtcca agtatgtgaa cttcctctac ctggcctccc actacgagaa
    11221 gctcaagggc agccccgagg acaacgagca gaagcagctg ttcgtcgagc agcacaagca
    11281 ttacctcgac gagatcattg agcagatttc cgagttctcc aagcgcgtga tcctggccga
    11341 cgcgaatctg gataaggtcc tctccgcgta caacaagcac cgcgacaagc caatcaggga
    11401 gcaggctgag aatatcattc atctcttcac cctgacgaac ctcggcgccc ctgctgcttt
    11461 caagtacttc gacacaacta tcgatcgcaa gaggtacaca agcactaagg aggtcctgga
    11521 cgcgaccctc atccaccagt cgattaccgg cctctacgag acgcgcatcg acctgtctca
    11581 gctcgggggc gacaagcggc cagcggcgac gaagaaggcg gggcaggcga agaagaagaa
    11641 gtgagctcag agctttcgtt cgtatcatcg gtttcgacaa cgttcgtcaa gttcaatgca
    11701 tcagtttcat tgcgcacaca ccagaatcct actgagtttg agtattatgg cattgggaaa
    11761 actgtttttc ttgtaccatt tgttgtgctt gtaatttact gtgtttttta ttcggttttc
    11821 gctatcgaac tgtgaaatgg aaatggatgg agaagagtta atgaatgata tggtcctttt
    11881 gttcattctc aaattaatat tatttgtttt ttctcttatt tgttgtgtgt tgaatttgaa
    11941 attataagag atatgcaaac attttgtttt gagtaaaaat gtgtcaaatc gtggcctcta
    12001 atgaccgaag ttaatatgag gagtaaaaca cttgtagttg taccattatg cttattcact
    12061 aggcaacaaa tatattttca gacctagaaa agctgcaaat gttactgaat acaagtatgt
    12121 cctcttgtgt tttagacatt tatgaacttt cctttatgta attttccaga atccttgtca
    12181 gattctaatc attgctttat aattatagtt atactcatgg atttgtagtt gagtatgaaa
    12241 atatttttta atgcatttta tgacttgcca attgttccgg aactctagat aagcttaccg
    12301 gaaagggcga attcgcaact ttgtatacaa aagttgcccc atggcgttcc ctctagataa
    12361 cgcaggatcc tatttacttt aaatttttct tatggctcag cctgtgatgg ataactgaat
    12421 caaacaaatg gcgtctgggt ttaagaacat ctgttttggc tatgttggac gaaacaagtg
    12481 aacttttagg atcaacttcc gtttatatac ggagcttata tcgagcaata agataagtgg
    12541 gctttttatg taatttaatg ggctatcgtc catatattca ctaataccca tgcccagtac
    12601 ccatgtatgc gtttcatata agctcctaat ttctcccaca tcgctcaaat ctaaacaaat
    12661 cttgttgtat atataacact gagggagcac cattggtcag agaccggcgc cgctacaggg
    12721 cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
    12781 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
    12841 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga
    12901 ctcactatag ggcgaattgg gtaccgggcc ccccctcgag gtcctccagc ttttgttccc
    12961 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
    13021 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
    13081 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
    13141 accggtggtc tctgttttag agctagaaat agcaagttaa aataaggcta gtccgttatc
    13201 aacttgaaaa agtggcaccg agtcggtgct tttttttccc tttccttttt tctttttttt
    13261 gccataaact taaatttgta tatcgatcat tgtagatatt gaaaacctag aacaaaccaa
    13321 catccatgtg aatgtctttc atgactgatt tagagataat tcttgaattt tggaactaga
    13381 atctataatg agcctaaatt aaaacattgt gaactagtaa gggcgaattc gacccagctt
    13441 tcttgtacaa agtggttcga taattcttaa ttaactagtt ctagagcggc cgccaccgcg
    13501 gtggagctcg aatttccccg atcgttcaaa catttggcaa taaagtttct taagattgaa
    13561 tcctgttgcc ggtcttgcga tgattatcat ataatttctg ttgaattacg ttaagcatgt
    13621 aataattaac atgtaatgca tgacgttatt tatgagatgg gtttttatga ttagagtccc
    13681 gcaattatac atttaatacg cgatagaaaa caaaatatag cgcgcaaact aggataaatt
    13741 atcgcgcgcg gtgtcatcta tgttactgaa ttcgtaatca tggtcataga agcttgcatg
    13801 cctgcaggtc gactctagag gatccccggg taccgagctc gaattcgtaa tcatgtcata
    13861 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
    13921 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
    13981 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
    14041 acgcgcgggg agaggcggtt tgcgtattgg ctagagcagc ttgccaacat ggtggagcac
    14101 gacactctcg tctactccaa gaatatcaaa gatacagtct cagaagacca aagggctatt
    14161 gagacttttc aacaaagggt aatatcggga aacctcctcg gattccattg cccagctatc
    14221 tgtcacttca tcaaaaggac agtagaaaag gaaggtggca cctacaaatg ccatcattgc
    14281 gataaaggaa aggctatcgt tcaagatgcc tctgccgaca gtggtcccaa agatggaccc
    14341 ccacccacga ggagcatcgt ggaaaaagaa gacgttccaa ccacgtcttc aaagcaagtg
    14401 gattgatgtg aacatggtgg agcacgacac tctcgtctac tccaagaata tcaaagatac
    14461 agtctcagaa gaccaaaggg ctattgagac ttttcaacaa agggtaatat cgggaaacct
    14521 cctcggattc cattgcccag ctatctgtca cttcatcaaa aggacagtag aaaaggaagg
    14581 tggcacctac aaatgccatc attgcgataa aggaaaggct atcgttcaag atgcctctgc
    14641 cgacagtggt cccaaagatg gacccccacc cacgaggagc atcgtggaaa aagaagacgt
    14701 tccaaccacg tcttcaaagc aagtggattg atgtgatatc tccactgacg taagggatga
    14761 cgcacaatcc cactatcctt cgcaagaccc ttcctctata taaggaagtt catttcattt
    14821 ggagaggaca cgctgaaatc accagtctct ctctacaaat ctatctctct cgagctttcg
    14881 cagatctgtc gatcgaccat ggggattgaa caagatggat tgcacgcagg ttctccggcc
    14941 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
    15001 gccgccgtgt ttcggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
    15061 tccggtgccc tgaatgaact ccaggacgag gcagcgcggc tatcgtggct ggccacgacg
    15121 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
    15181 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
    15241 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
    15301 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
    15361 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
    15421 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cacatggcga tgcctgcttg
    15481 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
    15541 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
    15601 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
    15661 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcggatcga
    15721 tcctctagct agagtcgatc gacaagctcg agtttctcca taataatgtg tgagtagttc
    15781 ccagataagg gaattagggt tcctataggg tttcgctcat gtgttgagca tataagaaac
    15841 ccttagtatg tatttgtatt tgtaaaatac ttctatcaat aaaatttcta attcctaaaa
    15901 ccaaaatcca gtactaaaat ccagatcccc cgaattaatt cggcgttaat tcagtacatt
    15961 aaaaacgtcc gcaatgtgtt attaagttgt ctaagcgtca atttgtttac accacaatat
    16021 atcctgcca
//
