LOCUS       sequence-452221-       16668 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T-DNA vector for SpCas9-D10A based A-to-G base editing for dicot 
            plants; NGG PAM; SpCas9D10A-ABE 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 16668)
  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 16668)
  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: pYPQAT33
FEATURES             Location/Qualifiers
     source          1..16668
                     /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             7394..7414
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8015..8080
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             8087..8107
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(8132..8155,8156..8158,8159..12232)
                     /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             12145..12153
                     /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             12233..12280
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     protein_bind    12955..12975
                     /gene="mutant version of attB"
                     /label=attB5
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     CDS             complement(13336..13641)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGAGL"
     primer_bind     13506..13522
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     13532..13551
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        13532..13550
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(13598..13618)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(13598..13616)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(13637..13653)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    13661..13677
                     /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(13685..13691,13692..13709,13710..13715))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13730..13751
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_RNA        13793..13868
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(14071..14095)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    14429..14485
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(14494..14510)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    14518..14534
                     /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(14542..14548,14549..14566,14567..14572))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    14587..14608
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        14799..15475
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     CDS             15538..16335
                     /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(15595..15614)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     16205..16224
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    16392..16566
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    16644..16668
                     /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 ggctccgatg aaacggacag ccgacggaag
     7381 cgagttcgag tcaccaaaga agaagcggaa agtctctgag gtggagtttt cccacgagta
     7441 ctggatgaga catgccctga ccctggccaa gagggcacgg gatgagaggg aggtgcctgt
     7501 gggagccgtg ctggtgctga acaatagagt gatcggcgag ggctggaaca gagccatcgg
     7561 cctgcacgac ccaacagccc atgccgaaat tatggccctg agacagggcg gcctggtcat
     7621 gcagaactac agactgattg acgccaccct gtacgtgaca ttcgagcctt gcgtgatgtg
     7681 cgccggcgcc atgatccact ctaggatcgg ccgcgtggtg tttggcgtga ggaactcaaa
     7741 aagaggcgcc gcaggctccc tgatgaacgt gctgaactac cccggcatga atcaccgcgt
     7801 cgaaattacc gagggaatcc tggcagatga atgtgccgcc ctgctgtgcg atttctatcg
     7861 gatgcctaga caggtgttca atgctcagaa gaaggcccag agctccatca actccggagg
     7921 atctagcgga ggctcctctg gctctgagac acctggcaca agcgagagcg caacacctga
     7981 aagcagcggg ggcagcagcg gggggtcaac catggattac aaggaccacg acggggatta
     8041 caaggaccac gacattgatt acaaggatga tgatgacaag atggctccga agaagaagag
     8101 gaaggttggc atccacgggg tgccagctgc tgacaagaag tactcgatcg gcctcgctat
     8161 tgggactaac tctgttggct gggccgtgat caccgacgag tacaaggtgc cctcaaagaa
     8221 gttcaaggtc ctgggcaaca ccgatcggca ttccatcaag aagaatctca ttggcgctct
     8281 cctgttcgac agcggcgaga cggctgaggc tacgcggctc aagcgcaccg cccgcaggcg
     8341 gtacacgcgc aggaagaatc gcatctgcta cctgcaggag attttctcca acgagatggc
     8401 gaaggttgac gattctttct tccacaggct ggaggagtca ttcctcgtgg aggaggataa
     8461 gaagcacgag cggcatccaa tcttcggcaa cattgtcgac gaggttgcct accacgagaa
     8521 gtaccctacg atctaccatc tgcggaagaa gctcgtggac tccacagata aggcggacct
     8581 ccgcctgatc tacctcgctc tggcccacat gattaagttc aggggccatt tcctgatcga
     8641 gggggatctc aacccggaca atagcgatgt tgacaagctg ttcatccagc tcgtgcagac
     8701 gtacaaccag ctcttcgagg agaaccccat taatgcgtca ggcgtcgacg cgaaggctat
     8761 cctgtccgct aggctctcga agtctcggcg cctcgagaac ctgatcgccc agctgccggg
     8821 cgagaagaag aacggcctgt tcgggaatct cattgcgctc agcctggggc tcacgcccaa
     8881 cttcaagtcg aatttcgatc tcgctgagga cgccaagctg cagctctcca aggacacata
     8941 cgacgatgac ctggataacc tcctggccca gatcggcgat cagtacgcgg acctgttcct
     9001 cgctgccaag aatctgtcgg acgccatcct cctgtctgat attctcaggg tgaacaccga
     9061 gattacgaag gctccgctct cagcctccat gatcaagcgc tacgacgagc accatcagga
     9121 tctgaccctc ctgaaggcgc tggtcaggca gcagctcccc gagaagtaca aggagatctt
     9181 cttcgatcag tcgaagaacg gctacgctgg gtacattgac ggcggggcct ctcaggagga
     9241 gttctacaag ttcatcaagc cgattctgga gaagatggac ggcacggagg agctgctggt
     9301 gaagctcaat cgcgaggacc tcctgaggaa gcagcggaca ttcgataacg gcagcatccc
     9361 acaccagatt catctcgggg agctgcacgc tatcctgagg aggcaggagg acttctaccc
     9421 tttcctcaag gataaccgcg agaagatcga gaagattctg actttcagga tcccgtacta
     9481 cgtcggccca ctcgctaggg gcaactcccg cttcgcttgg atgacccgca agtcagagga
     9541 gacgatcacg ccgtggaact tcgaggaggt ggtcgacaag ggcgctagcg ctcagtcgtt
     9601 catcgagagg atgacgaatt tcgacaagaa cctgccaaat gagaaggtgc tccctaagca
     9661 ctcgctcctg tacgagtact tcacagtcta caacgagctg actaaggtga agtatgtgac
     9721 cgagggcatg aggaagccgg ctttcctgtc tggggagcag aagaaggcca tcgtggacct
     9781 cctgttcaag accaaccgga aggtcacggt taagcagctc aaggaggact acttcaagaa
     9841 gattgagtgc ttcgattcgg tcgagatctc tggcgttgag gaccgcttca acgcctccct
     9901 ggggacctac cacgatctcc tgaagatcat taaggataag gacttcctgg acaacgagga
     9961 gaatgaggat atcctcgagg acattgtgct gacactcact ctgttcgagg accgggagat
    10021 gatcgaggag cgcctgaaga cttacgccca tctcttcgat gacaaggtca tgaagcagct
    10081 caagaggagg aggtacaccg gctgggggag gctgagcagg aagctcatca acggcattcg
    10141 ggacaagcag tccgggaaga cgatcctcga cttcctgaag agcgatggct tcgcgaaccg
    10201 caatttcatg cagctgattc acgatgacag cctcacattc aaggaggata tccagaaggc
    10261 tcaggtgagc ggccaggggg actcgctgca cgagcatatc gcgaacctcg ctggctcgcc
    10321 agctatcaag aaggggattc tgcagaccgt gaaggttgtg gacgagctgg tgaaggtcat
    10381 gggcaggcac aagcctgaga acatcgtcat tgagatggcc cgggagaatc agaccacgca
    10441 gaagggccag aagaactcac gcgagaggat gaagaggatc gaggagggca ttaaggagct
    10501 ggggtcccag atcctcaagg agcacccggt ggagaacacg cagctgcaga atgagaagct
    10561 ctacctgtac tacctccaga atggccgcga tatgtatgtg gaccaggagc tggatattaa
    10621 caggctcagc gattacgacg tcgatcatat cgttccacag tcattcctga aggatgactc
    10681 cattgacaac aaggtcctca ccaggtcgga caagaaccgg ggcaagtctg ataatgttcc
    10741 ttcagaggag gtcgttaaga agatgaagaa ctactggcgc cagctcctga atgccaagct
    10801 gatcacgcag cggaagttcg ataacctcac aaaggctgag aggggcgggc tctctgagct
    10861 ggacaaggcg ggcttcatca agaggcagct ggtcgagaca cggcagatca ctaagcacgt
    10921 tgcgcagatt ctcgactcac ggatgaacac taagtacgat gagaatgaca agctgatccg
    10981 cgaggtgaag gtcatcaccc tgaagtcaaa gctcgtctcc gacttcagga aggatttcca
    11041 gttctacaag gttcgggaga tcaacaatta ccaccatgcc catgacgcgt acctgaacgc
    11101 ggtggtcggc acagctctga tcaagaagta cccaaagctc gagagcgagt tcgtgtacgg
    11161 ggactacaag gtttacgatg tgaggaagat gatcgccaag tcggagcagg agattggcaa
    11221 ggctaccgcc aagtacttct tctactctaa cattatgaat ttcttcaaga cagagatcac
    11281 tctggccaat ggcgagatcc ggaagcgccc cctcatcgag acgaacggcg agacggggga
    11341 gatcgtgtgg gacaagggca gggatttcgc gaccgtcagg aaggttctct ccatgccaca
    11401 agtgaatatc gtcaagaaga cagaggtcca gactggcggg ttctctaagg agtcaattct
    11461 gcctaagcgg aacagcgaca agctcatcgc ccgcaagaag gactgggatc cgaagaagta
    11521 cggcgggttc gacagcccca ctgtggccta ctcggtcctg gttgtggcga aggttgagaa
    11581 gggcaagtcc aagaagctca agagcgtgaa ggagctgctg gggatcacga ttatggagcg
    11641 ctccagcttc gagaagaacc cgatcgattt cctggaggcg aagggctaca aggaggtgaa
    11701 gaaggacctg atcattaagc tccccaagta ctcactcttc gagctggaga acggcaggaa
    11761 gcggatgctg gcttccgctg gcgagctgca gaaggggaac gagctggctc tgccgtccaa
    11821 gtatgtgaac ttcctctacc tggcctccca ctacgagaag ctcaagggca gccccgagga
    11881 caacgagcag aagcagctgt tcgtcgagca gcacaagcat tacctcgacg agatcattga
    11941 gcagatttcc gagttctcca agcgcgtgat cctggccgac gcgaatctgg ataaggtcct
    12001 ctccgcgtac aacaagcacc gcgacaagcc aatcagggag caggctgaga atatcattca
    12061 tctcttcacc ctgacgaacc tcggcgcccc tgctgctttc aagtacttcg acacaactat
    12121 cgatcgcaag aggtacacaa gcactaagga ggtcctggac gcgaccctca tccaccagtc
    12181 gattaccggc ctctacgaga cgcgcatcga cctgtctcag ctcgggggcg acaagcggcc
    12241 agcggcgacg aagaaggcgg ggcaggcgaa gaagaagaag tgagctcaga gctttcgttc
    12301 gtatcatcgg tttcgacaac gttcgtcaag ttcaatgcat cagtttcatt gcgcacacac
    12361 cagaatccta ctgagtttga gtattatggc attgggaaaa ctgtttttct tgtaccattt
    12421 gttgtgcttg taatttactg tgttttttat tcggttttcg ctatcgaact gtgaaatgga
    12481 aatggatgga gaagagttaa tgaatgatat ggtccttttg ttcattctca aattaatatt
    12541 atttgttttt tctcttattt gttgtgtgtt gaatttgaaa ttataagaga tatgcaaaca
    12601 ttttgttttg agtaaaaatg tgtcaaatcg tggcctctaa tgaccgaagt taatatgagg
    12661 agtaaaacac ttgtagttgt accattatgc ttattcacta ggcaacaaat atattttcag
    12721 acctagaaaa gctgcaaatg ttactgaata caagtatgtc ctcttgtgtt ttagacattt
    12781 atgaactttc ctttatgtaa ttttccagaa tccttgtcag attctaatca ttgctttata
    12841 attatagtta tactcatgga tttgtagttg agtatgaaaa tattttttaa tgcattttat
    12901 gacttgccaa ttgttccgga actctagata agcttaccgg aaagggcgaa ttcgcaactt
    12961 tgtatacaaa agttgcccca tggcgttccc tctagataac gcaggatcct atttacttta
    13021 aatttttctt atggctcagc ctgtgatgga taactgaatc aaacaaatgg cgtctgggtt
    13081 taagaacatc tgttttggct atgttggacg aaacaagtga acttttagga tcaacttccg
    13141 tttatatacg gagcttatat cgagcaataa gataagtggg ctttttatgt aatttaatgg
    13201 gctatcgtcc atatattcac taatacccat gcccagtacc catgtatgcg tttcatataa
    13261 gctcctaatt tctcccacat cgctcaaatc taaacaaatc ttgttgtata tataacactg
    13321 agggagcacc attggtcaga gaccggcgcc gctacagggc gcgtcccatt cgccattcag
    13381 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
    13441 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
    13501 acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac tcactatagg gcgaattggg
    13561 taccgggccc cccctcgagg tcctccagct tttgttccct ttagtgaggg ttaattgcgc
    13621 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
    13681 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
    13741 aactcacatt aattgcgttg cgctcactgc ccgctttcca ccggtggtct ctgttttaga
    13801 gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga
    13861 gtcggtgctt ttttttccct ttcctttttt cttttttttg ccataaactt aaatttgtat
    13921 atcgatcatt gtagatattg aaaacctaga acaaaccaac atccatgtga atgtctttca
    13981 tgactgattt agagataatt cttgaatttt ggaactagaa tctataatga gcctaaatta
    14041 aaacattgtg aactagtaag ggcgaattcg acccagcttt cttgtacaaa gtggttcgat
    14101 aattcttaat taactagttc tagagcggcc gccaccgcgg tggagctcga atttccccga
    14161 tcgttcaaac atttggcaat aaagtttctt aagattgaat cctgttgccg gtcttgcgat
    14221 gattatcata taatttctgt tgaattacgt taagcatgta ataattaaca tgtaatgcat
    14281 gacgttattt atgagatggg tttttatgat tagagtcccg caattataca tttaatacgc
    14341 gatagaaaac aaaatatagc gcgcaaacta ggataaatta tcgcgcgcgg tgtcatctat
    14401 gttactgaat tcgtaatcat ggtcatagaa gcttgcatgc ctgcaggtcg actctagagg
    14461 atccccgggt accgagctcg aattcgtaat catgtcatag ctgtttcctg tgtgaaattg
    14521 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
    14581 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
    14641 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
    14701 gcgtattggc tagagcagct tgccaacatg gtggagcacg acactctcgt ctactccaag
    14761 aatatcaaag atacagtctc agaagaccaa agggctattg agacttttca acaaagggta
    14821 atatcgggaa acctcctcgg attccattgc ccagctatct gtcacttcat caaaaggaca
    14881 gtagaaaagg aaggtggcac ctacaaatgc catcattgcg ataaaggaaa ggctatcgtt
    14941 caagatgcct ctgccgacag tggtcccaaa gatggacccc cacccacgag gagcatcgtg
    15001 gaaaaagaag acgttccaac cacgtcttca aagcaagtgg attgatgtga acatggtgga
    15061 gcacgacact ctcgtctact ccaagaatat caaagataca gtctcagaag accaaagggc
    15121 tattgagact tttcaacaaa gggtaatatc gggaaacctc ctcggattcc attgcccagc
    15181 tatctgtcac ttcatcaaaa ggacagtaga aaaggaaggt ggcacctaca aatgccatca
    15241 ttgcgataaa ggaaaggcta tcgttcaaga tgcctctgcc gacagtggtc ccaaagatgg
    15301 acccccaccc acgaggagca tcgtggaaaa agaagacgtt ccaaccacgt cttcaaagca
    15361 agtggattga tgtgatatct ccactgacgt aagggatgac gcacaatccc actatccttc
    15421 gcaagaccct tcctctatat aaggaagttc atttcatttg gagaggacac gctgaaatca
    15481 ccagtctctc tctacaaatc tatctctctc gagctttcgc agatctgtcg atcgaccatg
    15541 gggattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
    15601 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt tcggctgtca
    15661 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactc
    15721 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
    15781 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
    15841 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
    15901 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
    15961 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
    16021 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
    16081 ggcgaggatc tcgtcgtgac acatggcgat gcctgcttgc cgaatatcat ggtggaaaat
    16141 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
    16201 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
    16261 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
    16321 gacgagttct tctgagcggg actctggggt tcggatcgat cctctagcta gagtcgatcg
    16381 acaagctcga gtttctccat aataatgtgt gagtagttcc cagataaggg aattagggtt
    16441 cctatagggt ttcgctcatg tgttgagcat ataagaaacc cttagtatgt atttgtattt
    16501 gtaaaatact tctatcaata aaatttctaa ttcctaaaac caaaatccag tactaaaatc
    16561 cagatccccc gaattaattc ggcgttaatt cagtacatta aaaacgtccg caatgtgtta
    16621 ttaagttgtc taagcgtcaa tttgtttaca ccacaatata tcctgcca
//
