LOCUS       sequence-452579-       17185 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T-DNA vector for SpCas9-D10A based A-to-G base editing for monocot 
            plants; NGG PAM; SpCas9D10A-ABE was driven by ZmUbi1 and the sgRNA 
            was driven by OsU3 promoter; BASTA for plants selection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17185)
  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 17185)
  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: pYPQAT35
FEATURES             Location/Qualifiers
     source          1..17185
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    283..307
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     510..526
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        542..2539
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     protein_bind    2582..2606
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2646..2666
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3267..3332
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3339..3359
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3384..3407,3408..3410,3411..7484)
                     /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             7397..7405
                     /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             7485..7532
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     protein_bind    8207..8227
                     /gene="mutant version of attB"
                     /label=attB5
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        8264..8641
                     /label=OsU3 promoter
                     /note="Oryza sativa (rice) snRNA U3 promoter"
     CDS             complement(8612..8944)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGAGLCHG
                     SSAQLF"
     primer_bind     8809..8825
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8835..8854
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8835..8853
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8901..8921)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(8901..8919)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8940..8956)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8964..8980
                     /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(8988..8994,8995..9012,9013..9018))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9033..9054
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_RNA        9096..9171
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(9260..9284)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(9352..9368)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    9376..9392
                     /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(9400..9406,9407..9424,9425..9430))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9445..9466
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        9657..10333
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     10270..10290
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             10378..10929
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     polyA_signal    10936..11110
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    11188..11212
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             11637..12431
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      12518..13106
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     13007..13026
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     13260..13277
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    13292..13432
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(13358..13377)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     13518..13540
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      13776..13970
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(14036..15109)
                     /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(15538..16167)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
ORIGIN
        1 ctactttatg cgactaaaac acgcgacaag aaaacgccag gaaaagggca gggcggcagc
       61 ctgtcgcgta acttaggact tgtgcgacat gtcgttttca gaagacggct gcactgaacg
      121 tcagaagccg actgcactat agcagcggag gggttggatc aaagtacttt gatcccgagg
      181 ggaaccctgt ggttggcatg cacatacaaa tggacgaacg gataaacctt ttcacgccct
      241 tttaaatatc cgattattct aataaacgct cttttctctt aggtttaccc gccaatatat
      301 cctgtcaaac actgatagtt taaactgaag gcgggaaacg acaatctgat ccaagctcaa
      361 gctgctctag cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
      421 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
      481 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca agcttgcatg
      541 cctgcagtgc agcgtgaccc ggtcgtgccc ctctctagag ataatgagca ttgcatgtct
      601 aagttataaa aaattaccac atattttttt tgtcacactt gtttgaagtg cagtttatct
      661 atctttatac atatatttaa actttactct acgaataata taatctatag tactacaata
      721 atatcagtgt tttagagaat catataaatg aacagttaga catggtctaa aggacaattg
      781 agtattttga caacaggact ctacagtttt atctttttag tgtgcatgtg ttctcctttt
      841 tttttgcaaa tagcttcacc tatataatac ttcatccatt ttattagtac atccatttag
      901 ggtttagggt taatggtttt tatagactaa tttttttagt acatctattt tattctattt
      961 tagcctctaa attaagaaaa ctaaaactct attttagttt ttttatttaa taatttagat
     1021 ataaaataga ataaaataaa gtgactaaaa attaaacaaa taccctttaa gaaattaaaa
     1081 aaactaagga aacatttttc ttgtttcgag tagataatgc cagcctgtta aacgccgtcg
     1141 acgagtctaa cggacaccaa ccagcgaacc agcagcgtcg cgtcgggcca agcgaagcag
     1201 acggcacggc atctctgtcg ctgcctctgg acccctctcg agagttccgc tccaccgttg
     1261 gacttgctcc gctgtcggca tccagaaatt gcgtggcgga gcggcagacg tgagccggca
     1321 cggcaggcgg cctcctcctc ctctcacggc accggcagct acgggggatt cctttcccac
     1381 cgctccttcg ctttcccttc ctcgcccgcc gtaataaata gacaccccct ccacaccctc
     1441 tttccccaac ctcgtgttgt tcggagcgca cacacacaca accagatctc ccccaaatcc
     1501 acccgtcggc acctccgctt caaggtacgc cgctcgtccy cccccccccc cccctctcta
     1561 ccttctctag atcggcgttc cggtccatgg ttagggcccg gtagttctac ttctgttcat
     1621 gtttgtgtta gatccgtgtt tgtgttagat ccgtgctgct agcgttcgta cacggatgcg
     1681 acctgtacgt cagacacgtt ctgattgcta acttgccagt gtttctcttt ggggaatcct
     1741 gggatggctc tagccgttcc gcagacggga tcgatttcat gatttttttt gtttcgttgc
     1801 atagggtttg gtttgccctt ttcctttatt tcaatatatg ccgtgcactt gtttgtcggg
     1861 tcatcttttc atgctttttt ttgtcttggt tgtgatgatg tggtctggtt gggcggtcgt
     1921 tctagatcgg agtagaatta attctgtttc aaactacctg gtggatttat taattttgga
     1981 tctgtatgtg tgtgccatac atattcatag ttacgaattg aagatgatgg atggaaatat
     2041 cgatctagga taggtataca tgttgatgcg ggttttactg atgcatatac agagatgctt
     2101 tttgttcgct tggttgtgat gatgtggtgt ggttgggcgg tcgttcattc gttctagatc
     2161 ggagtagaat actgtttcaa actacctggt gtatttatta attttggaac tgtatgtgtg
     2221 tgtcatacat cttcatagtt acgagtttaa gatggatgga aatatcgatc taggataggt
     2281 atacatgttg atgtgggttt tactgatgca tatacatgat ggcatatgca gcatctattc
     2341 atatgctcta accttgagta cctatctatt ataataaaca agtatgtttt ataattattt
     2401 tgatcttgat atacttggat gatggcatat gcagcagcta tatgtggatt tttttagccc
     2461 tgccttcata cgctatttat ttgcttggta ctgtttcttt tgtcgatgct caccctgttg
     2521 tttggtgtta cttctgcagg tcgactctag aggatcccct cgaggcgcgc caagctatca
     2581 aacaagtttg tacaaaaaag caggctccga tgaaacggac agccgacgga agcgagttcg
     2641 agtcaccaaa gaagaagcgg aaagtctctg aggtggagtt ttcccacgag tactggatga
     2701 gacatgccct gaccctggcc aagagggcac gggatgagag ggaggtgcct gtgggagccg
     2761 tgctggtgct gaacaataga gtgatcggcg agggctggaa cagagccatc ggcctgcacg
     2821 acccaacagc ccatgccgaa attatggccc tgagacaggg cggcctggtc atgcagaact
     2881 acagactgat tgacgccacc ctgtacgtga cattcgagcc ttgcgtgatg tgcgccggcg
     2941 ccatgatcca ctctaggatc ggccgcgtgg tgtttggcgt gaggaactca aaaagaggcg
     3001 ccgcaggctc cctgatgaac gtgctgaact accccggcat gaatcaccgc gtcgaaatta
     3061 ccgagggaat cctggcagat gaatgtgccg ccctgctgtg cgatttctat cggatgccta
     3121 gacaggtgtt caatgctcag aagaaggccc agagctccat caactccgga ggatctagcg
     3181 gaggctcctc tggctctgag acacctggca caagcgagag cgcaacacct gaaagcagcg
     3241 ggggcagcag cggggggtca accatggatt acaaggacca cgacggggat tacaaggacc
     3301 acgacattga ttacaaggat gatgatgaca agatggctcc gaagaagaag aggaaggttg
     3361 gcatccacgg ggtgccagct gctgacaaga agtactcgat cggcctcgct attgggacta
     3421 actctgttgg ctgggccgtg atcaccgacg agtacaaggt gccctcaaag aagttcaagg
     3481 tcctgggcaa caccgatcgg cattccatca agaagaatct cattggcgct ctcctgttcg
     3541 acagcggcga gacggctgag gctacgcggc tcaagcgcac cgcccgcagg cggtacacgc
     3601 gcaggaagaa tcgcatctgc tacctgcagg agattttctc caacgagatg gcgaaggttg
     3661 acgattcttt cttccacagg ctggaggagt cattcctcgt ggaggaggat aagaagcacg
     3721 agcggcatcc aatcttcggc aacattgtcg acgaggttgc ctaccacgag aagtacccta
     3781 cgatctacca tctgcggaag aagctcgtgg actccacaga taaggcggac ctccgcctga
     3841 tctacctcgc tctggcccac atgattaagt tcaggggcca tttcctgatc gagggggatc
     3901 tcaacccgga caatagcgat gttgacaagc tgttcatcca gctcgtgcag acgtacaacc
     3961 agctcttcga ggagaacccc attaatgcgt caggcgtcga cgcgaaggct atcctgtccg
     4021 ctaggctctc gaagtctcgg cgcctcgaga acctgatcgc ccagctgccg ggcgagaaga
     4081 agaacggcct gttcgggaat ctcattgcgc tcagcctggg gctcacgccc aacttcaagt
     4141 cgaatttcga tctcgctgag gacgccaagc tgcagctctc caaggacaca tacgacgatg
     4201 acctggataa cctcctggcc cagatcggcg atcagtacgc ggacctgttc ctcgctgcca
     4261 agaatctgtc ggacgccatc ctcctgtctg atattctcag ggtgaacacc gagattacga
     4321 aggctccgct ctcagcctcc atgatcaagc gctacgacga gcaccatcag gatctgaccc
     4381 tcctgaaggc gctggtcagg cagcagctcc ccgagaagta caaggagatc ttcttcgatc
     4441 agtcgaagaa cggctacgct gggtacattg acggcggggc ctctcaggag gagttctaca
     4501 agttcatcaa gccgattctg gagaagatgg acggcacgga ggagctgctg gtgaagctca
     4561 atcgcgagga cctcctgagg aagcagcgga cattcgataa cggcagcatc ccacaccaga
     4621 ttcatctcgg ggagctgcac gctatcctga ggaggcagga ggacttctac cctttcctca
     4681 aggataaccg cgagaagatc gagaagattc tgactttcag gatcccgtac tacgtcggcc
     4741 cactcgctag gggcaactcc cgcttcgctt ggatgacccg caagtcagag gagacgatca
     4801 cgccgtggaa cttcgaggag gtggtcgaca agggcgctag cgctcagtcg ttcatcgaga
     4861 ggatgacgaa tttcgacaag aacctgccaa atgagaaggt gctccctaag cactcgctcc
     4921 tgtacgagta cttcacagtc tacaacgagc tgactaaggt gaagtatgtg accgagggca
     4981 tgaggaagcc ggctttcctg tctggggagc agaagaaggc catcgtggac ctcctgttca
     5041 agaccaaccg gaaggtcacg gttaagcagc tcaaggagga ctacttcaag aagattgagt
     5101 gcttcgattc ggtcgagatc tctggcgttg aggaccgctt caacgcctcc ctggggacct
     5161 accacgatct cctgaagatc attaaggata aggacttcct ggacaacgag gagaatgagg
     5221 atatcctcga ggacattgtg ctgacactca ctctgttcga ggaccgggag atgatcgagg
     5281 agcgcctgaa gacttacgcc catctcttcg atgacaaggt catgaagcag ctcaagagga
     5341 ggaggtacac cggctggggg aggctgagca ggaagctcat caacggcatt cgggacaagc
     5401 agtccgggaa gacgatcctc gacttcctga agagcgatgg cttcgcgaac cgcaatttca
     5461 tgcagctgat tcacgatgac agcctcacat tcaaggagga tatccagaag gctcaggtga
     5521 gcggccaggg ggactcgctg cacgagcata tcgcgaacct cgctggctcg ccagctatca
     5581 agaaggggat tctgcagacc gtgaaggttg tggacgagct ggtgaaggtc atgggcaggc
     5641 acaagcctga gaacatcgtc attgagatgg cccgggagaa tcagaccacg cagaagggcc
     5701 agaagaactc acgcgagagg atgaagagga tcgaggaggg cattaaggag ctggggtccc
     5761 agatcctcaa ggagcacccg gtggagaaca cgcagctgca gaatgagaag ctctacctgt
     5821 actacctcca gaatggccgc gatatgtatg tggaccagga gctggatatt aacaggctca
     5881 gcgattacga cgtcgatcat atcgttccac agtcattcct gaaggatgac tccattgaca
     5941 acaaggtcct caccaggtcg gacaagaacc ggggcaagtc tgataatgtt ccttcagagg
     6001 aggtcgttaa gaagatgaag aactactggc gccagctcct gaatgccaag ctgatcacgc
     6061 agcggaagtt cgataacctc acaaaggctg agaggggcgg gctctctgag ctggacaagg
     6121 cgggcttcat caagaggcag ctggtcgaga cacggcagat cactaagcac gttgcgcaga
     6181 ttctcgactc acggatgaac actaagtacg atgagaatga caagctgatc cgcgaggtga
     6241 aggtcatcac cctgaagtca aagctcgtct ccgacttcag gaaggatttc cagttctaca
     6301 aggttcggga gatcaacaat taccaccatg cccatgacgc gtacctgaac gcggtggtcg
     6361 gcacagctct gatcaagaag tacccaaagc tcgagagcga gttcgtgtac ggggactaca
     6421 aggtttacga tgtgaggaag atgatcgcca agtcggagca ggagattggc aaggctaccg
     6481 ccaagtactt cttctactct aacattatga atttcttcaa gacagagatc actctggcca
     6541 atggcgagat ccggaagcgc cccctcatcg agacgaacgg cgagacgggg gagatcgtgt
     6601 gggacaaggg cagggatttc gcgaccgtca ggaaggttct ctccatgcca caagtgaata
     6661 tcgtcaagaa gacagaggtc cagactggcg ggttctctaa ggagtcaatt ctgcctaagc
     6721 ggaacagcga caagctcatc gcccgcaaga aggactggga tccgaagaag tacggcgggt
     6781 tcgacagccc cactgtggcc tactcggtcc tggttgtggc gaaggttgag aagggcaagt
     6841 ccaagaagct caagagcgtg aaggagctgc tggggatcac gattatggag cgctccagct
     6901 tcgagaagaa cccgatcgat ttcctggagg cgaagggcta caaggaggtg aagaaggacc
     6961 tgatcattaa gctccccaag tactcactct tcgagctgga gaacggcagg aagcggatgc
     7021 tggcttccgc tggcgagctg cagaagggga acgagctggc tctgccgtcc aagtatgtga
     7081 acttcctcta cctggcctcc cactacgaga agctcaaggg cagccccgag gacaacgagc
     7141 agaagcagct gttcgtcgag cagcacaagc attacctcga cgagatcatt gagcagattt
     7201 ccgagttctc caagcgcgtg atcctggccg acgcgaatct ggataaggtc ctctccgcgt
     7261 acaacaagca ccgcgacaag ccaatcaggg agcaggctga gaatatcatt catctcttca
     7321 ccctgacgaa cctcggcgcc cctgctgctt tcaagtactt cgacacaact atcgatcgca
     7381 agaggtacac aagcactaag gaggtcctgg acgcgaccct catccaccag tcgattaccg
     7441 gcctctacga gacgcgcatc gacctgtctc agctcggggg cgacaagcgg ccagcggcga
     7501 cgaagaaggc ggggcaggcg aagaagaaga agtgagctca gagctttcgt tcgtatcatc
     7561 ggtttcgaca acgttcgtca agttcaatgc atcagtttca ttgcgcacac accagaatcc
     7621 tactgagttt gagtattatg gcattgggaa aactgttttt cttgtaccat ttgttgtgct
     7681 tgtaatttac tgtgtttttt attcggtttt cgctatcgaa ctgtgaaatg gaaatggatg
     7741 gagaagagtt aatgaatgat atggtccttt tgttcattct caaattaata ttatttgttt
     7801 tttctcttat ttgttgtgtg ttgaatttga aattataaga gatatgcaaa cattttgttt
     7861 tgagtaaaaa tgtgtcaaat cgtggcctct aatgaccgaa gttaatatga ggagtaaaac
     7921 acttgtagtt gtaccattat gcttattcac taggcaacaa atatattttc agacctagaa
     7981 aagctgcaaa tgttactgaa tacaagtatg tcctcttgtg ttttagacat ttatgaactt
     8041 tcctttatgt aattttccag aatccttgtc agattctaat cattgcttta taattatagt
     8101 tatactcatg gatttgtagt tgagtatgaa aatatttttt aatgcatttt atgacttgcc
     8161 aattgttccg gaactctaga taagcttacc ggaaagggcg aattcgcaac tttgtataca
     8221 aaagttgccc catggcgttc cctctagata acgcaggatc ctaaagggat ctttaaacat
     8281 acgaacagat cacttaaagt tcttctgaag caacttaaag ttatcaggca tgcatggatc
     8341 ttggaggaat cagatgtgca gtcagggacc atagcacagg acaggcgtct tctactggtg
     8401 ctaccagcaa atgctggaag ccgggaacac tgggtacgtt ggaaaccacg tgatgtggag
     8461 taagataaac tgtaggagaa aagcatttcg tagtgggcca tgaagccttt caggacatgt
     8521 attgcagtat gggccggccc attacgcaat tggacgacaa caaagactag tattagtacc
     8581 acctcggcta tccacataga tcaaagctgg tttaaaagag ttgtgcagat gatccgtggc
     8641 agagaccggc gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg
     8701 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     8761 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     8821 ccagtgagcg cgcgtaatac gactcactat agggcgaatt gggtaccggg ccccccctcg
     8881 aggtcctcca gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg
     8941 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     9001 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     9061 ttgcgctcac tgcccgcttt ccaccggtgg tctctgtttt agagctagaa atagcaagtt
     9121 aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg ctttttttgt
     9181 cccttcgaag ggcaattctg cagatatcca tcacactggc ggccgctcga ggtcgagggt
     9241 atcgataagc ttgaattcga cccagctttc ttgtacaaag tggttcgata attcttaatt
     9301 aactagttct agagcggccg ccaccgcggt ggagctcgaa ttcgtaatca tgtcatagct
     9361 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     9421 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     9481 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     9541 cgcggggaga ggcggtttgc gtattggcta gagcagcttg ccaacatggt ggagcacgac
     9601 actctcgtct actccaagaa tatcaaagat acagtctcag aagaccaaag ggctattgag
     9661 acttttcaac aaagggtaat atcgggaaac ctcctcggat tccattgccc agctatctgt
     9721 cacttcatca aaaggacagt agaaaaggaa ggtggcacct acaaatgcca tcattgcgat
     9781 aaaggaaagg ctatcgttca agatgcctct gccgacagtg gtcccaaaga tggaccccca
     9841 cccacgagga gcatcgtgga aaaagaagac gttccaacca cgtcttcaaa gcaagtggat
     9901 tgatgtgaac atggtggagc acgacactct cgtctactcc aagaatatca aagatacagt
     9961 ctcagaagac caaagggcta ttgagacttt tcaacaaagg gtaatatcgg gaaacctcct
    10021 cggattccat tgcccagcta tctgtcactt catcaaaagg acagtagaaa aggaaggtgg
    10081 cacctacaaa tgccatcatt gcgataaagg aaaggctatc gttcaagatg cctctgccga
    10141 cagtggtccc aaagatggac ccccacccac gaggagcatc gtggaaaaag aagacgttcc
    10201 aaccacgtct tcaaagcaag tggattgatg tgatatctcc actgacgtaa gggatgacgc
    10261 acaatcccac tatccttcgc aagacccttc ctctatataa ggaagttcat ttcatttgga
    10321 gaggacacgc tgaaatcacc agtctctctc tacaaatcta tctctctcga gtctaccatg
    10381 agcccagaac gacgcccggc cgacatccgc cgtgccaccg aggcggacat gccggcggtc
    10441 tgcaccatcg tcaaccacta catcgagaca agcacggtca acttccgtac cgagccgcag
    10501 gaaccgcagg agtggacgga cgacctcgtc cgtctgcggg agcgctatcc ctggctcgtc
    10561 gccgaggtgg acggcgaggt cgccggcatc gcctacgcgg gcccctggaa ggcacgcaac
    10621 gcctacgact ggacggccga gtcgaccgtg tacgtctccc cccgccacca gcggacggga
    10681 ctgggctcca cgctctacac ccacctgctg aagtccctgg aggcacaggg cttcaagagc
    10741 gtggtcgctg tcatcgggct gcccaacgac ccgagcgtgc gcatgcacga ggcgctcgga
    10801 tatgcccccc gcggcatgct gcgggcggcc ggcttcaagc acgggaactg gcatgacgtg
    10861 ggtttctggc agctggactt cagcctgccg gtaccgcccc gtccggtcct gcccgtcacc
    10921 gagatttgac tcgagtttct ccataataat gtgtgagtag ttcccagata agggaattag
    10981 ggttcctata gggtttcgct catgtgttga gcatataaga aacccttagt atgtatttgt
    11041 atttgtaaaa tacttctatc aataaaattt ctaattccta aaaccaaaat ccagtactaa
    11101 aatccagatc ccccgaatta attcggcgtt aattcagtac attaaaaacg tccgcaatgt
    11161 gttattaagt tgtctaagcg tcaatttgtt tacaccacaa tatatcctgc caccagccag
    11221 ccaacagctc cccgaccggc agctcggcac aaaatcacca ctcgatacag gcagcccatc
    11281 agtccgggac ggcgtcagcg ggagagccgt tgtaaggcgg cagactttgc tcatgttacc
    11341 gatgctattc ggaagaacgg caactaagct gccgggtttg aaacacggat gatctcgcgg
    11401 agggtagcat gttgattgta acgatgacag agcgttgctg cctgtgatca ccgcggtttc
    11461 aaaatcggct ccgtcgatac tatgttatac gccaactttg aaaacaactt tgaaaaagct
    11521 gttttctggt atttaaggtt ttagaatgca aggaacagtg aattggagtt cgtcttgtta
    11581 taattagctt cttggggtat ctttaaatac tgtagaaaag aggaaggaaa taataaatgg
    11641 ctaaaatgag aatatcaccg gaattgaaaa aactgatcga aaaataccgc tgcgtaaaag
    11701 atacggaagg aatgtctcct gctaaggtat ataagctggt gggagaaaat gaaaacctat
    11761 atttaaaaat gacggacagc cggtataaag ggaccaccta tgatgtggaa cgggaaaagg
    11821 acatgatgct atggctggaa ggaaagctgc ctgttccaaa ggtcctgcac tttgaacggc
    11881 atgatggctg gagcaatctg ctcatgagtg aggccgatgg cgtcctttgc tcggaagagt
    11941 atgaagatga acaaagccct gaaaagatta tcgagctgta tgcggagtgc atcaggctct
    12001 ttcactccat cgacatatcg gattgtccct atacgaatag cttagacagc cgcttagccg
    12061 aattggatta cttactgaat aacgatctgg ccgatgtgga ttgcgaaaac tgggaagaag
    12121 acactccatt taaagatccg cgcgagctgt atgatttttt aaagacggaa aagcccgaag
    12181 aggaacttgt cttttcccac ggcgacctgg gagacagcaa catctttgtg aaagatggca
    12241 aagtaagtgg ctttattgat cttgggagaa gcggcagggc ggacaagtgg tatgacattg
    12301 ccttctgcgt ccggtcgatc agggaggata tcggggaaga acagtatgtc gagctatttt
    12361 ttgacttact ggggatcaag cctgattggg agaaaataaa atattatatt ttactggatg
    12421 aattgtttta gtacctagaa tgcatgacca aaatccctta acgtgagttt tcgttccact
    12481 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
    12541 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
    12601 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
    12661 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
    12721 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
    12781 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
    12841 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
    12901 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
    12961 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
    13021 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
    13081 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
    13141 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
    13201 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
    13261 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
    13321 tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat
    13381 agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca
    13441 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
    13501 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
    13561 acgcgcgagg cagggtgcct tgatgtgggc gccggcggtc gagtggcgac ggcgcggctt
    13621 gtccgcgccc tggtagattg cctggccgta ggccagccat ttttgagcgg ccagcggccg
    13681 cgataggccg acgcgaagcg gcggggcgta gggagcgcag cgaccgaagg gtaggcgctt
    13741 tttgcagctc ttcggctgtg cgctggccag acagttatgc acaggccagg cgggttttaa
    13801 gagttttaat aagttttaaa gagttttagg cggaaaaatc gccttttttc tcttttatat
    13861 cagtcactta catgtgtgac cggttcccaa tgtacggctt tgggttccca atgtacgggt
    13921 tccggttccc aatgtacggc tttgggttcc caatgtacgt gctatccaca ggaaacagac
    13981 cttttcgacc tttttcccct gctagggcaa tttgccctag catctgctcc gtacattagg
    14041 aaccggcgga tgcttcgccc tcgatcaggt tgcggtagcg catgactagg atcgggccag
    14101 cctgccccgc ctcctccttc aaatcgtact ccggcaggtc atttgacccg atcagcttgc
    14161 gcacggtgaa acagaacttc ttgaactctc cggcgctgcc actgcgttcg tagatcgtct
    14221 tgaacaacca tctggcttct gccttgcctg cggcgcggcg tgccaggcgg tagagaaaac
    14281 ggccgatgcc gggatcgatc aaaaagtaat cggggtgaac cgtcagcacg tccgggttct
    14341 tgccttctgt gatctcgcgg tacatccaat cagctagctc gatctcgatg tactccggcc
    14401 gcccggtttc gctctttacg atcttgtagc ggctaatcaa ggcttcaccc tcggataccg
    14461 tcaccaggcg gccgttcttg gccttcttcg tacgctgcat ggcaacgtgc gtggtgttta
    14521 accgaatgca ggtttctacc aggtcgtctt tctgctttcc gccatcggct cgccggcaga
    14581 acttgagtac gtccgcaacg tgtggacgga acacgcggcc gggcttgtct cccttccctt
    14641 cccggtatcg gttcatggat tcggttagat gggaaaccgc catcagtacc aggtcgtaat
    14701 cccacacact ggccatgccg gccggccctg cggaaacctc tacgtgcccg tctggaagct
    14761 cgtagcggat cacctcgcca gctcgtcggt cacgcttcga cagacggaaa acggccacgt
    14821 ccatgatgct gcgactatcg cgggtgccca cgtcatagag catcggaacg aaaaaatctg
    14881 gttgctcgtc gcccttgggc ggcttcctaa tcgacggcgc accggctgcc ggcggttgcc
    14941 gggattcttt gcggattcga tcagcggccg cttgccacga ttcaccgggg cgtgcttctg
    15001 cctcgatgcg ttgccgctgg gcggcctgcg cggccttcaa cttctccacc aggtcatcac
    15061 ccagcgccgc gccgatttgt accgggccgg atggtttgcg accgctcacg ccgattcctc
    15121 gggcttgggg gttccagtgc cattgcaggg ccggcaggca acccagccgc ttacgcctgg
    15181 ccaaccgccc gttcctccac acatggggca ttccacggcg tcggtgcctg gttgttcttg
    15241 attttccatg ccgcctcctt tagccgctaa aattcatcta ctcatttatt catttgctca
    15301 tttactctgg tagctgcgcg atgtattcag atagcagctc ggtaatggtc ttgccttggc
    15361 gtaccgcgta catcttcagc ttggtgtgat cctccgccgg caactgaaag ttgacccgct
    15421 tcatggctgg cgtgtctgcc aggctggcca acgttgcagc cttgctgctg cgtgcgctcg
    15481 gacggccggc acttagcgtg tttgtgcttt tgctcatttt ctctttacct cattaactca
    15541 aatgagtttt gatttaattt cagcggccag cgcctggacc tcgcgggcag cgtcgccctc
    15601 gggttctgat tcaagaacgg ttgtgccggc ggcggcagtg cctgggtagc tcacgcgctg
    15661 cgtgatacgg gactcaagaa tgggcagctc gtacccggcc agcgcctcgg caacctcacc
    15721 gccgatgcgc gtgcctttga tcgcccgcga cacgacaaag gccgcttgta gccttccatc
    15781 cgtgacctca atgcgctgct taaccagctc caccaggtcg gcggtggccc atatgtcgta
    15841 agggcttggc tgcaccggaa tcagcacgaa gtcggctgcc ttgatcgcgg acacagccaa
    15901 gtccgccgcc tggggcgctc cgtcgatcac tacgaagtcg cgccggccga tggccttcac
    15961 gtcgcggtca atcgtcgggc ggtcgatgcc gacaacggtt agcggttgat cttcccgcac
    16021 ggccgcccaa tcgcgggcac tgccctgggg atcggaatcg actaacagaa catcggcccc
    16081 ggcgagttgc agggcgcggg ctagatgggt tgcgatggtc gtcttgcctg acccgccttt
    16141 ctggttaagt acagcgataa ccttcatgcg ttccccttgc gtatttgttt atttactcat
    16201 cgcatcatat acgcagcgac cgcatgacgc aagctgtttt actcaaatac acatcacctt
    16261 tttagacggc ggcgctcggt ttcttcagcg gccaagctgg ccggccaggc cgccagcttg
    16321 gcatcagaca aaccggccag gatttcatgc agccgcacgg ttgagacgtg cgcgggcggc
    16381 tcgaacacgt acccggccgc gatcatctcc gcctcgatct cttcggtaat gaaaaacggt
    16441 tcgtcctggc cgtcctggtg cggtttcatg cttgttcctc ttggcgttca ttctcggcgg
    16501 ccgccagggc gtcggcctcg gtcaatgcgt cctcacggaa ggcaccgcgc cgcctggcct
    16561 cggtgggcgt cacttcctcg ctgcgctcaa gtgcgcggta cagggtcgag cgatgcacgc
    16621 caagcagtgc agccgcctct ttcacggtgc ggccttcctg gtcgatcagc tcgcgggcgt
    16681 gcgcgatctg tgccggggtg agggtagggc gggggccaaa cttcacgcct cgggccttgg
    16741 cggcctcgcg cccgctccgg gtgcggtcga tgattaggga acgctcgaac tcggcaatgc
    16801 cggcgaacac ggtcaacacc atgcggccgg ccggcgtggt ggtgtcggcc cacggctctg
    16861 ccaggctacg caggcccgcg ccggcctcct ggatgcgctc ggcaatgtcc agtaggtcgc
    16921 gggtgctgcg ggccaggcgg tctagcctgg tcactgtcac aacgtcgcca gggcgtaggt
    16981 ggtcaagcat cctggccagc tccgggcggt cgcgcctggt gccggtgatc ttctcggaaa
    17041 acagcttggt gcagccggcc gcgtgcagtt cggcccgttg gttggtcaag tcctggtcgt
    17101 cggtgctgac gcgggcatag cccagcaggc cagcggcggc gctcttgttc atggcgtaat
    17161 gtctccggtt ctagtcgcaa gtatt
//
