LOCUS       sequence-452296-       16621 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T-DNA vector for A3A/Y130F-CBE_V01 based C-to-T base editing for 
            dicot plants; NGG PAM; wide working window; A3A/Y130F-CBE_V01 was 
            driven by 2x35s and the sgRNA was driven by AtU3 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16621)
  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 16621)
  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: pYPQAT15
FEATURES             Location/Qualifiers
     source          1..16621
                     /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             8021..8086
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             8093..8113
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(8138..8161,8162..8164,8165..12238)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     RBS             12151..12159
                     /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             12239..12286
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             12308..12556
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             12569..12589
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      12611..12863
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    12908..12928
                     /gene="mutant version of attB"
                     /label=attB5
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     CDS             complement(13289..13594)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGAGL"
     primer_bind     13459..13475
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     13485..13504
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        13485..13503
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(13551..13571)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(13551..13569)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(13590..13606)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    13614..13630
                     /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(13638..13644,13645..13662,13663..13668))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13683..13704
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_RNA        13746..13821
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(14024..14048)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    14382..14438
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(14447..14463)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    14471..14487
                     /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(14495..14501,14502..14519,14520..14525))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    14540..14561
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        14752..15428
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     CDS             15491..16288
                     /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(15548..15567)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     16158..16177
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    16345..16519
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    16597..16621
                     /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 tggaggcctc
     7381 tccggcttca ggtcctcgcc atctcatgga cccccatatc tttacctcca atttcaataa
     7441 cggcattggc aggcacaaga catacctctg ttatgaggtc gagcggctcg ataacggaac
     7501 ctcagtgaag atggaccaac acaggggttt ccttcataac caagcgaaaa atctcctttg
     7561 cggtttctat gggcggcacg cggagctgag attcttggat ttggtcccga gcctccagct
     7621 ggatcccgcg caaatatacc gcgtcacttg gttcatatca tggagcccgt gctttagctg
     7681 ggggtgtgcc ggtgaggttc gcgcgttcct ccaggaaaac acccatgttc gcctccggat
     7741 tttcgcagcg aggattttcg actatgatcc actttacaaa gaggccctcc aaatgcttag
     7801 agatgctggg gctcaagttt caattatgac atacgacgag tttaagcatt gttgggacac
     7861 ttttgtcgat catcagggtt gtcccttcca gccgtgggat ggtcttgatg agcattcgca
     7921 agctttgagt ggcaggcttc gcgcgatact tcagaaccag ggtaactcag gttctgagac
     7981 gcctggcaca agtgagtcag caacacccga gtccaccatg gattacaagg accacgacgg
     8041 ggattacaag gaccacgaca ttgattacaa ggatgatgat gacaagatgg ctccgaagaa
     8101 gaagaggaag gttggcatcc acggggtgcc agctgctgac aagaagtact cgatcggcct
     8161 cgctattggg actaactctg ttggctgggc cgtgatcacc gacgagtaca aggtgccctc
     8221 aaagaagttc aaggtcctgg gcaacaccga tcggcattcc atcaagaaga atctcattgg
     8281 cgctctcctg ttcgacagcg gcgagacggc tgaggctacg cggctcaagc gcaccgcccg
     8341 caggcggtac acgcgcagga agaatcgcat ctgctacctg caggagattt tctccaacga
     8401 gatggcgaag gttgacgatt ctttcttcca caggctggag gagtcattcc tcgtggagga
     8461 ggataagaag cacgagcggc atccaatctt cggcaacatt gtcgacgagg ttgcctacca
     8521 cgagaagtac cctacgatct accatctgcg gaagaagctc gtggactcca cagataaggc
     8581 ggacctccgc ctgatctacc tcgctctggc ccacatgatt aagttcaggg gccatttcct
     8641 gatcgagggg gatctcaacc cggacaatag cgatgttgac aagctgttca tccagctcgt
     8701 gcagacgtac aaccagctct tcgaggagaa ccccattaat gcgtcaggcg tcgacgcgaa
     8761 ggctatcctg tccgctaggc tctcgaagtc tcggcgcctc gagaacctga tcgcccagct
     8821 gccgggcgag aagaagaacg gcctgttcgg gaatctcatt gcgctcagcc tggggctcac
     8881 gcccaacttc aagtcgaatt tcgatctcgc tgaggacgcc aagctgcagc tctccaagga
     8941 cacatacgac gatgacctgg ataacctcct ggcccagatc ggcgatcagt acgcggacct
     9001 gttcctcgct gccaagaatc tgtcggacgc catcctcctg tctgatattc tcagggtgaa
     9061 caccgagatt acgaaggctc cgctctcagc ctccatgatc aagcgctacg acgagcacca
     9121 tcaggatctg accctcctga aggcgctggt caggcagcag ctccccgaga agtacaagga
     9181 gatcttcttc gatcagtcga agaacggcta cgctgggtac attgacggcg gggcctctca
     9241 ggaggagttc tacaagttca tcaagccgat tctggagaag atggacggca cggaggagct
     9301 gctggtgaag ctcaatcgcg aggacctcct gaggaagcag cggacattcg ataacggcag
     9361 catcccacac cagattcatc tcggggagct gcacgctatc ctgaggaggc aggaggactt
     9421 ctaccctttc ctcaaggata accgcgagaa gatcgagaag attctgactt tcaggatccc
     9481 gtactacgtc ggcccactcg ctaggggcaa ctcccgcttc gcttggatga cccgcaagtc
     9541 agaggagacg atcacgccgt ggaacttcga ggaggtggtc gacaagggcg ctagcgctca
     9601 gtcgttcatc gagaggatga cgaatttcga caagaacctg ccaaatgaga aggtgctccc
     9661 taagcactcg ctcctgtacg agtacttcac agtctacaac gagctgacta aggtgaagta
     9721 tgtgaccgag ggcatgagga agccggcttt cctgtctggg gagcagaaga aggccatcgt
     9781 ggacctcctg ttcaagacca accggaaggt cacggttaag cagctcaagg aggactactt
     9841 caagaagatt gagtgcttcg attcggtcga gatctctggc gttgaggacc gcttcaacgc
     9901 ctccctgggg acctaccacg atctcctgaa gatcattaag gataaggact tcctggacaa
     9961 cgaggagaat gaggatatcc tcgaggacat tgtgctgaca ctcactctgt tcgaggaccg
    10021 ggagatgatc gaggagcgcc tgaagactta cgcccatctc ttcgatgaca aggtcatgaa
    10081 gcagctcaag aggaggaggt acaccggctg ggggaggctg agcaggaagc tcatcaacgg
    10141 cattcgggac aagcagtccg ggaagacgat cctcgacttc ctgaagagcg atggcttcgc
    10201 gaaccgcaat ttcatgcagc tgattcacga tgacagcctc acattcaagg aggatatcca
    10261 gaaggctcag gtgagcggcc agggggactc gctgcacgag catatcgcga acctcgctgg
    10321 ctcgccagct atcaagaagg ggattctgca gaccgtgaag gttgtggacg agctggtgaa
    10381 ggtcatgggc aggcacaagc ctgagaacat cgtcattgag atggcccggg agaatcagac
    10441 cacgcagaag ggccagaaga actcacgcga gaggatgaag aggatcgagg agggcattaa
    10501 ggagctgggg tcccagatcc tcaaggagca cccggtggag aacacgcagc tgcagaatga
    10561 gaagctctac ctgtactacc tccagaatgg ccgcgatatg tatgtggacc aggagctgga
    10621 tattaacagg ctcagcgatt acgacgtcga tcatatcgtt ccacagtcat tcctgaagga
    10681 tgactccatt gacaacaagg tcctcaccag gtcggacaag aaccggggca agtctgataa
    10741 tgttccttca gaggaggtcg ttaagaagat gaagaactac tggcgccagc tcctgaatgc
    10801 caagctgatc acgcagcgga agttcgataa cctcacaaag gctgagaggg gcgggctctc
    10861 tgagctggac aaggcgggct tcatcaagag gcagctggtc gagacacggc agatcactaa
    10921 gcacgttgcg cagattctcg actcacggat gaacactaag tacgatgaga atgacaagct
    10981 gatccgcgag gtgaaggtca tcaccctgaa gtcaaagctc gtctccgact tcaggaagga
    11041 tttccagttc tacaaggttc gggagatcaa caattaccac catgcccatg acgcgtacct
    11101 gaacgcggtg gtcggcacag ctctgatcaa gaagtaccca aagctcgaga gcgagttcgt
    11161 gtacggggac tacaaggttt acgatgtgag gaagatgatc gccaagtcgg agcaggagat
    11221 tggcaaggct accgccaagt acttcttcta ctctaacatt atgaatttct tcaagacaga
    11281 gatcactctg gccaatggcg agatccggaa gcgccccctc atcgagacga acggcgagac
    11341 gggggagatc gtgtgggaca agggcaggga tttcgcgacc gtcaggaagg ttctctccat
    11401 gccacaagtg aatatcgtca agaagacaga ggtccagact ggcgggttct ctaaggagtc
    11461 aattctgcct aagcggaaca gcgacaagct catcgcccgc aagaaggact gggatccgaa
    11521 gaagtacggc gggttcgaca gccccactgt ggcctactcg gtcctggttg tggcgaaggt
    11581 tgagaagggc aagtccaaga agctcaagag cgtgaaggag ctgctgggga tcacgattat
    11641 ggagcgctcc agcttcgaga agaacccgat cgatttcctg gaggcgaagg gctacaagga
    11701 ggtgaagaag gacctgatca ttaagctccc caagtactca ctcttcgagc tggagaacgg
    11761 caggaagcgg atgctggctt ccgctggcga gctgcagaag gggaacgagc tggctctgcc
    11821 gtccaagtat gtgaacttcc tctacctggc ctcccactac gagaagctca agggcagccc
    11881 cgaggacaac gagcagaagc agctgttcgt cgagcagcac aagcattacc tcgacgagat
    11941 cattgagcag atttccgagt tctccaagcg cgtgatcctg gccgacgcga atctggataa
    12001 ggtcctctcc gcgtacaaca agcaccgcga caagccaatc agggagcagg ctgagaatat
    12061 cattcatctc ttcaccctga cgaacctcgg cgcccctgct gctttcaagt acttcgacac
    12121 aactatcgat cgcaagaggt acacaagcac taaggaggtc ctggacgcga ccctcatcca
    12181 ccagtcgatt accggcctct acgagacgcg catcgacctg tctcagctcg ggggcgacaa
    12241 gcggccagcg gcgacgaaga aggcggggca ggcgaagaag aagaagggag acggatcagg
    12301 cgggtcaaca aacctcagtg acatcataga gaaggaaact ggtaagcaac tggttattca
    12361 agagagcatt ctgatgctcc ctgaggaagt cgaggaagtt ataggaaaca agcctgagag
    12421 cgatatactc gtgcatacgg cgtatgacga gagcacggat gaaaatgtga tgctcttgac
    12481 cagcgatgcg ccagaatata aaccctgggc actggttatt caggactcta acggggaaaa
    12541 taaaatcaag atgctctcag gtggctcccc aaagaagaaa cgcaaggttt agtggcggcc
    12601 gccgacgtcc gatcgttcaa acatttggca ataaagtttc ttaagattga atcctgttgc
    12661 cggtcttgcg atgattatca tataatttct gttgaattac gttaagcatg taataattaa
    12721 catgtaatgc atgacgttat ttatgagatg ggtttttatg attagagtcc cgcaattata
    12781 catttaatac gcgatagaaa acaaaatata gcgcgcaaac taggataaat tatcgcgcgc
    12841 ggtgtcatct atgttactag atcgggaatt gatcccccct cgacagcttc cggaaagggc
    12901 gaattcgcaa ctttgtatac aaaagttgcc ccatggcgtt ccctctagat aacgcaggat
    12961 cctatttact ttaaattttt cttatggctc agcctgtgat ggataactga atcaaacaaa
    13021 tggcgtctgg gtttaagaac atctgttttg gctatgttgg acgaaacaag tgaactttta
    13081 ggatcaactt ccgtttatat acggagctta tatcgagcaa taagataagt gggcttttta
    13141 tgtaatttaa tgggctatcg tccatatatt cactaatacc catgcccagt acccatgtat
    13201 gcgtttcata taagctccta atttctccca catcgctcaa atctaaacaa atcttgttgt
    13261 atatataaca ctgagggagc accattggtc agagaccggc gccgctacag ggcgcgtccc
    13321 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
    13381 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
    13441 tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat
    13501 agggcgaatt gggtaccggg ccccccctcg aggtcctcca gcttttgttc cctttagtga
    13561 gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
    13621 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
    13681 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccaccggtgg
    13741 tctctgtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa
    13801 aaagtggcac cgagtcggtg cttttttttc cctttccttt tttctttttt ttgccataaa
    13861 cttaaatttg tatatcgatc attgtagata ttgaaaacct agaacaaacc aacatccatg
    13921 tgaatgtctt tcatgactga tttagagata attcttgaat tttggaacta gaatctataa
    13981 tgagcctaaa ttaaaacatt gtgaactagt aagggcgaat tcgacccagc tttcttgtac
    14041 aaagtggttc gataattctt aattaactag ttctagagcg gccgccaccg cggtggagct
    14101 cgaatttccc cgatcgttca aacatttggc aataaagttt cttaagattg aatcctgttg
    14161 ccggtcttgc gatgattatc atataatttc tgttgaatta cgttaagcat gtaataatta
    14221 acatgtaatg catgacgtta tttatgagat gggtttttat gattagagtc ccgcaattat
    14281 acatttaata cgcgatagaa aacaaaatat agcgcgcaaa ctaggataaa ttatcgcgcg
    14341 cggtgtcatc tatgttactg aattcgtaat catggtcata gaagcttgca tgcctgcagg
    14401 tcgactctag aggatccccg ggtaccgagc tcgaattcgt aatcatgtca tagctgtttc
    14461 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
    14521 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
    14581 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
    14641 ggagaggcgg tttgcgtatt ggctagagca gcttgccaac atggtggagc acgacactct
    14701 cgtctactcc aagaatatca aagatacagt ctcagaagac caaagggcta ttgagacttt
    14761 tcaacaaagg gtaatatcgg gaaacctcct cggattccat tgcccagcta tctgtcactt
    14821 catcaaaagg acagtagaaa aggaaggtgg cacctacaaa tgccatcatt gcgataaagg
    14881 aaaggctatc gttcaagatg cctctgccga cagtggtccc aaagatggac ccccacccac
    14941 gaggagcatc gtggaaaaag aagacgttcc aaccacgtct tcaaagcaag tggattgatg
    15001 tgaacatggt ggagcacgac actctcgtct actccaagaa tatcaaagat acagtctcag
    15061 aagaccaaag ggctattgag acttttcaac aaagggtaat atcgggaaac ctcctcggat
    15121 tccattgccc agctatctgt cacttcatca aaaggacagt agaaaaggaa ggtggcacct
    15181 acaaatgcca tcattgcgat aaaggaaagg ctatcgttca agatgcctct gccgacagtg
    15241 gtcccaaaga tggaccccca cccacgagga gcatcgtgga aaaagaagac gttccaacca
    15301 cgtcttcaaa gcaagtggat tgatgtgata tctccactga cgtaagggat gacgcacaat
    15361 cccactatcc ttcgcaagac ccttcctcta tataaggaag ttcatttcat ttggagagga
    15421 cacgctgaaa tcaccagtct ctctctacaa atctatctct ctcgagcttt cgcagatctg
    15481 tcgatcgacc atggggattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
    15541 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
    15601 gtttcggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
    15661 cctgaatgaa ctccaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
    15721 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
    15781 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
    15841 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
    15901 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
    15961 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
    16021 gcgcatgccc gacggcgagg atctcgtcgt gacacatggc gatgcctgct tgccgaatat
    16081 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
    16141 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
    16201 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
    16261 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcggatc gatcctctag
    16321 ctagagtcga tcgacaagct cgagtttctc cataataatg tgtgagtagt tcccagataa
    16381 gggaattagg gttcctatag ggtttcgctc atgtgttgag catataagaa acccttagta
    16441 tgtatttgta tttgtaaaat acttctatca ataaaatttc taattcctaa aaccaaaatc
    16501 cagtactaaa atccagatcc cccgaattaa ttcggcgtta attcagtaca ttaaaaacgt
    16561 ccgcaatgtg ttattaagtt gtctaagcgt caatttgttt acaccacaat atatcctgcc
    16621 a
//
