LOCUS       sequence-454173-       16546 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T-DNA vector for SpCas9 mediated mutagenesis for monocot plants; NGG
            PAM; Cas9 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 16546)
  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 16546)
  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: pYPQAT05
FEATURES             Location/Qualifiers
     source          1..16546
                     /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             2628..2693
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2700..2720
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2745..6845
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             2745..6845
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     RBS             6758..6766
                     /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             6846..6893
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     protein_bind    7568..7588
                     /gene="mutant version of attB"
                     /label=attB5
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        7625..8002
                     /label=OsU3 promoter
                     /note="Oryza sativa (rice) snRNA U3 promoter"
     CDS             complement(7973..8305)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGAGLCHG
                     SSAQLF"
     primer_bind     8170..8186
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8196..8215
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8196..8214
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8262..8282)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(8262..8280)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8301..8317)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8325..8341
                     /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(8349..8355,8356..8373,8374..8379))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8394..8415
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_RNA        8457..8532
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(8621..8645)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(8713..8729)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8737..8753
                     /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(8761..8767,8768..8785,8786..8791))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8806..8827
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        9018..9694
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     9631..9651
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             9739..10290
                     /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    10297..10471
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    10549..10573
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             10998..11792
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      11879..12467
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12368..12387
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12621..12638
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    12653..12793
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(12719..12738)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     12879..12901
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      13137..13331
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(13397..14470)
                     /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(14899..15528)
                     /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 cgctcgtcct 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 attcgccctt caccatggat tacaaggacc
     2641 acgacgggga ttacaaggac cacgacattg attacaagga tgatgatgac aagatggctc
     2701 cgaagaagaa gaggaaggtt ggcatccacg gggtgccagc tgctgacaag aagtactcga
     2761 tcggcctcga tattgggact aactctgttg gctgggccgt gatcaccgac gagtacaagg
     2821 tgccctcaaa gaagttcaag gtcctgggca acaccgatcg gcattccatc aagaagaatc
     2881 tcattggcgc tctcctgttc gacagcggcg agacggctga ggctacgcgg ctcaagcgca
     2941 ccgcccgcag gcggtacacg cgcaggaaga atcgcatctg ctacctgcag gagattttct
     3001 ccaacgagat ggcgaaggtt gacgattctt tcttccacag gctggaggag tcattcctcg
     3061 tggaggagga taagaagcac gagcggcatc caatcttcgg caacattgtc gacgaggttg
     3121 cctaccacga gaagtaccct acgatctacc atctgcggaa gaagctcgtg gactccacag
     3181 ataaggcgga cctccgcctg atctacctcg ctctggccca catgattaag ttcaggggcc
     3241 atttcctgat cgagggggat ctcaacccgg acaatagcga tgttgacaag ctgttcatcc
     3301 agctcgtgca gacgtacaac cagctcttcg aggagaaccc cattaatgcg tcaggcgtcg
     3361 acgcgaaggc tatcctgtcc gctaggctct cgaagtctcg gcgcctcgag aacctgatcg
     3421 cccagctgcc gggcgagaag aagaacggcc tgttcgggaa tctcattgcg ctcagcctgg
     3481 ggctcacgcc caacttcaag tcgaatttcg atctcgctga ggacgccaag ctgcagctct
     3541 ccaaggacac atacgacgat gacctggata acctcctggc ccagatcggc gatcagtacg
     3601 cggacctgtt cctcgctgcc aagaatctgt cggacgccat cctcctgtct gatattctca
     3661 gggtgaacac cgagattacg aaggctccgc tctcagcctc catgatcaag cgctacgacg
     3721 agcaccatca ggatctgacc ctcctgaagg cgctggtcag gcagcagctc cccgagaagt
     3781 acaaggagat cttcttcgat cagtcgaaga acggctacgc tgggtacatt gacggcgggg
     3841 cctctcagga ggagttctac aagttcatca agccgattct ggagaagatg gacggcacgg
     3901 aggagctgct ggtgaagctc aatcgcgagg acctcctgag gaagcagcgg acattcgata
     3961 acggcagcat cccacaccag attcatctcg gggagctgca cgctatcctg aggaggcagg
     4021 aggacttcta ccctttcctc aaggataacc gcgagaagat cgagaagatt ctgactttca
     4081 ggatcccgta ctacgtcggc ccactcgcta ggggcaactc ccgcttcgct tggatgaccc
     4141 gcaagtcaga ggagacgatc acgccgtgga acttcgagga ggtggtcgac aagggcgcta
     4201 gcgctcagtc gttcatcgag aggatgacga atttcgacaa gaacctgcca aatgagaagg
     4261 tgctccctaa gcactcgctc ctgtacgagt acttcacagt ctacaacgag ctgactaagg
     4321 tgaagtatgt gaccgagggc atgaggaagc cggctttcct gtctggggag cagaagaagg
     4381 ccatcgtgga cctcctgttc aagaccaacc ggaaggtcac ggttaagcag ctcaaggagg
     4441 actacttcaa gaagattgag tgcttcgatt cggtcgagat ctctggcgtt gaggaccgct
     4501 tcaacgcctc cctggggacc taccacgatc tcctgaagat cattaaggat aaggacttcc
     4561 tggacaacga ggagaatgag gatatcctcg aggacattgt gctgacactc actctgttcg
     4621 aggaccggga gatgatcgag gagcgcctga agacttacgc ccatctcttc gatgacaagg
     4681 tcatgaagca gctcaagagg aggaggtaca ccggctgggg gaggctgagc aggaagctca
     4741 tcaacggcat tcgggacaag cagtccggga agacgatcct cgacttcctg aagagcgatg
     4801 gcttcgcgaa ccgcaatttc atgcagctga ttcacgatga cagcctcaca ttcaaggagg
     4861 atatccagaa ggctcaggtg agcggccagg gggactcgct gcacgagcat atcgcgaacc
     4921 tcgctggctc gccagctatc aagaagggga ttctgcagac cgtgaaggtt gtggacgagc
     4981 tggtgaaggt catgggcagg cacaagcctg agaacatcgt cattgagatg gcccgggaga
     5041 atcagaccac gcagaagggc cagaagaact cacgcgagag gatgaagagg atcgaggagg
     5101 gcattaagga gctggggtcc cagatcctca aggagcaccc ggtggagaac acgcagctgc
     5161 agaatgagaa gctctacctg tactacctcc agaatggccg cgatatgtat gtggaccagg
     5221 agctggatat taacaggctc agcgattacg acgtcgatca tatcgttcca cagtcattcc
     5281 tgaaggatga ctccattgac aacaaggtcc tcaccaggtc ggacaagaac cggggcaagt
     5341 ctgataatgt tccttcagag gaggtcgtta agaagatgaa gaactactgg cgccagctcc
     5401 tgaatgccaa gctgatcacg cagcggaagt tcgataacct cacaaaggct gagaggggcg
     5461 ggctctctga gctggacaag gcgggcttca tcaagaggca gctggtcgag acacggcaga
     5521 tcactaagca cgttgcgcag attctcgact cacggatgaa cactaagtac gatgagaatg
     5581 acaagctgat ccgcgaggtg aaggtcatca ccctgaagtc aaagctcgtc tccgacttca
     5641 ggaaggattt ccagttctac aaggttcggg agatcaacaa ttaccaccat gcccatgacg
     5701 cgtacctgaa cgcggtggtc ggcacagctc tgatcaagaa gtacccaaag ctcgagagcg
     5761 agttcgtgta cggggactac aaggtttacg atgtgaggaa gatgatcgcc aagtcggagc
     5821 aggagattgg caaggctacc gccaagtact tcttctactc taacattatg aatttcttca
     5881 agacagagat cactctggcc aatggcgaga tccggaagcg ccccctcatc gagacgaacg
     5941 gcgagacggg ggagatcgtg tgggacaagg gcagggattt cgcgaccgtc aggaaggttc
     6001 tctccatgcc acaagtgaat atcgtcaaga agacagaggt ccagactggc gggttctcta
     6061 aggagtcaat tctgcctaag cggaacagcg acaagctcat cgcccgcaag aaggactggg
     6121 atccgaagaa gtacggcggg ttcgacagcc ccactgtggc ctactcggtc ctggttgtgg
     6181 cgaaggttga gaagggcaag tccaagaagc tcaagagcgt gaaggagctg ctggggatca
     6241 cgattatgga gcgctccagc ttcgagaaga acccgatcga tttcctggag gcgaagggct
     6301 acaaggaggt gaagaaggac ctgatcatta agctccccaa gtactcactc ttcgagctgg
     6361 agaacggcag gaagcggatg ctggcttccg ctggcgagct gcagaagggg aacgagctgg
     6421 ctctgccgtc caagtatgtg aacttcctct acctggcctc ccactacgag aagctcaagg
     6481 gcagccccga ggacaacgag cagaagcagc tgttcgtcga gcagcacaag cattacctcg
     6541 acgagatcat tgagcagatt tccgagttct ccaagcgcgt gatcctggcc gacgcgaatc
     6601 tggataaggt cctctccgcg tacaacaagc accgcgacaa gccaatcagg gagcaggctg
     6661 agaatatcat tcatctcttc accctgacga acctcggcgc ccctgctgct ttcaagtact
     6721 tcgacacaac tatcgatcgc aagaggtaca caagcactaa ggaggtcctg gacgcgaccc
     6781 tcatccacca gtcgattacc ggcctctacg agacgcgcat cgacctgtct cagctcgggg
     6841 gcgacaagcg gccagcggcg acgaagaagg cggggcaggc gaagaagaag aagtgagctc
     6901 agagctttcg ttcgtatcat cggtttcgac aacgttcgtc aagttcaatg catcagtttc
     6961 attgcgcaca caccagaatc ctactgagtt tgagtattat ggcattggga aaactgtttt
     7021 tcttgtacca tttgttgtgc ttgtaattta ctgtgttttt tattcggttt tcgctatcga
     7081 actgtgaaat ggaaatggat ggagaagagt taatgaatga tatggtcctt ttgttcattc
     7141 tcaaattaat attatttgtt ttttctctta tttgttgtgt gttgaatttg aaattataag
     7201 agatatgcaa acattttgtt ttgagtaaaa atgtgtcaaa tcgtggcctc taatgaccga
     7261 agttaatatg aggagtaaaa cacttgtagt tgtaccatta tgcttattca ctaggcaaca
     7321 aatatatttt cagacctaga aaagctgcaa atgttactga atacaagtat gtcctcttgt
     7381 gttttagaca tttatgaact ttcctttatg taattttcca gaatccttgt cagattctaa
     7441 tcattgcttt ataattatag ttatactcat ggatttgtag ttgagtatga aaatattttt
     7501 taatgcattt tatgacttgc caattgttcc ggaactctag ataagcttac cggaaagggc
     7561 gaattcgcaa ctttgtatac aaaagttgcc ccatggcgtt ccctctagat aacgcaggat
     7621 cctaaaggga tctttaaaca tacgaacaga tcacttaaag ttcttctgaa gcaacttaaa
     7681 gttatcaggc atgcatggat cttggaggaa tcagatgtgc agtcagggac catagcacag
     7741 gacaggcgtc ttctactggt gctaccagca aatgctggaa gccgggaaca ctgggtacgt
     7801 tggaaaccac gtgatgtgga gtaagataaa ctgtaggaga aaagcatttc gtagtgggcc
     7861 atgaagcctt tcaggacatg tattgcagta tgggccggcc cattacgcaa ttggacgaca
     7921 acaaagacta gtattagtac cacctcggct atccacatag atcaaagctg gtttaaaaga
     7981 gttgtgcaga tgatccgtgg cagagaccgg cgccgctaca gggcgcgtcc cattcgccat
     8041 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     8101 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
     8161 cacgacgttg taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat
     8221 tgggtaccgg gccccccctc gaggtcctcc agcttttgtt ccctttagtg agggttaatt
     8281 gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     8341 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     8401 agctaactca cattaattgc gttgcgctca ctgcccgctt tccaccggtg gtctctgttt
     8461 tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca
     8521 ccgagtcggt gctttttttg tcccttcgaa gggcaattct gcagatatcc atcacactgg
     8581 cggccgctcg aggtcgaggg tatcgataag cttgaattcg acccagcttt cttgtacaaa
     8641 gtggttcgat aattcttaat taactagttc tagagcggcc gccaccgcgg tggagctcga
     8701 attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     8761 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     8821 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     8881 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattggct agagcagctt
     8941 gccaacatgg tggagcacga cactctcgtc tactccaaga atatcaaaga tacagtctca
     9001 gaagaccaaa gggctattga gacttttcaa caaagggtaa tatcgggaaa cctcctcgga
     9061 ttccattgcc cagctatctg tcacttcatc aaaaggacag tagaaaagga aggtggcacc
     9121 tacaaatgcc atcattgcga taaaggaaag gctatcgttc aagatgcctc tgccgacagt
     9181 ggtcccaaag atggaccccc acccacgagg agcatcgtgg aaaaagaaga cgttccaacc
     9241 acgtcttcaa agcaagtgga ttgatgtgaa catggtggag cacgacactc tcgtctactc
     9301 caagaatatc aaagatacag tctcagaaga ccaaagggct attgagactt ttcaacaaag
     9361 ggtaatatcg ggaaacctcc tcggattcca ttgcccagct atctgtcact tcatcaaaag
     9421 gacagtagaa aaggaaggtg gcacctacaa atgccatcat tgcgataaag gaaaggctat
     9481 cgttcaagat gcctctgccg acagtggtcc caaagatgga cccccaccca cgaggagcat
     9541 cgtggaaaaa gaagacgttc caaccacgtc ttcaaagcaa gtggattgat gtgatatctc
     9601 cactgacgta agggatgacg cacaatccca ctatccttcg caagaccctt cctctatata
     9661 aggaagttca tttcatttgg agaggacacg ctgaaatcac cagtctctct ctacaaatct
     9721 atctctctcg agtctaccat gagcccagaa cgacgcccgg ccgacatccg ccgtgccacc
     9781 gaggcggaca tgccggcggt ctgcaccatc gtcaaccact acatcgagac aagcacggtc
     9841 aacttccgta ccgagccgca ggaaccgcag gagtggacgg acgacctcgt ccgtctgcgg
     9901 gagcgctatc cctggctcgt cgccgaggtg gacggcgagg tcgccggcat cgcctacgcg
     9961 ggcccctgga aggcacgcaa cgcctacgac tggacggccg agtcgaccgt gtacgtctcc
    10021 ccccgccacc agcggacggg actgggctcc acgctctaca cccacctgct gaagtccctg
    10081 gaggcacagg gcttcaagag cgtggtcgct gtcatcgggc tgcccaacga cccgagcgtg
    10141 cgcatgcacg aggcgctcgg atatgccccc cgcggcatgc tgcgggcggc cggcttcaag
    10201 cacgggaact ggcatgacgt gggtttctgg cagctggact tcagcctgcc ggtaccgccc
    10261 cgtccggtcc tgcccgtcac cgagatttga ctcgagtttc tccataataa tgtgtgagta
    10321 gttcccagat aagggaatta gggttcctat agggtttcgc tcatgtgttg agcatataag
    10381 aaacccttag tatgtatttg tatttgtaaa atacttctat caataaaatt tctaattcct
    10441 aaaaccaaaa tccagtacta aaatccagat cccccgaatt aattcggcgt taattcagta
    10501 cattaaaaac gtccgcaatg tgttattaag ttgtctaagc gtcaatttgt ttacaccaca
    10561 atatatcctg ccaccagcca gccaacagct ccccgaccgg cagctcggca caaaatcacc
    10621 actcgataca ggcagcccat cagtccggga cggcgtcagc gggagagccg ttgtaaggcg
    10681 gcagactttg ctcatgttac cgatgctatt cggaagaacg gcaactaagc tgccgggttt
    10741 gaaacacgga tgatctcgcg gagggtagca tgttgattgt aacgatgaca gagcgttgct
    10801 gcctgtgatc accgcggttt caaaatcggc tccgtcgata ctatgttata cgccaacttt
    10861 gaaaacaact ttgaaaaagc tgttttctgg tatttaaggt tttagaatgc aaggaacagt
    10921 gaattggagt tcgtcttgtt ataattagct tcttggggta tctttaaata ctgtagaaaa
    10981 gaggaaggaa ataataaatg gctaaaatga gaatatcacc ggaattgaaa aaactgatcg
    11041 aaaaataccg ctgcgtaaaa gatacggaag gaatgtctcc tgctaaggta tataagctgg
    11101 tgggagaaaa tgaaaaccta tatttaaaaa tgacggacag ccggtataaa gggaccacct
    11161 atgatgtgga acgggaaaag gacatgatgc tatggctgga aggaaagctg cctgttccaa
    11221 aggtcctgca ctttgaacgg catgatggct ggagcaatct gctcatgagt gaggccgatg
    11281 gcgtcctttg ctcggaagag tatgaagatg aacaaagccc tgaaaagatt atcgagctgt
    11341 atgcggagtg catcaggctc tttcactcca tcgacatatc ggattgtccc tatacgaata
    11401 gcttagacag ccgcttagcc gaattggatt acttactgaa taacgatctg gccgatgtgg
    11461 attgcgaaaa ctgggaagaa gacactccat ttaaagatcc gcgcgagctg tatgattttt
    11521 taaagacgga aaagcccgaa gaggaacttg tcttttccca cggcgacctg ggagacagca
    11581 acatctttgt gaaagatggc aaagtaagtg gctttattga tcttgggaga agcggcaggg
    11641 cggacaagtg gtatgacatt gccttctgcg tccggtcgat cagggaggat atcggggaag
    11701 aacagtatgt cgagctattt tttgacttac tggggatcaa gcctgattgg gagaaaataa
    11761 aatattatat tttactggat gaattgtttt agtacctaga atgcatgacc aaaatccctt
    11821 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
    11881 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
    11941 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
    12001 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
    12061 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
    12121 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
    12181 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
    12241 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
    12301 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
    12361 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
    12421 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
    12481 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
    12541 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
    12601 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
    12661 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta
    12721 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
    12781 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
    12841 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
    12901 gttttcaccg tcatcaccga aacgcgcgag gcagggtgcc ttgatgtggg cgccggcggt
    12961 cgagtggcga cggcgcggct tgtccgcgcc ctggtagatt gcctggccgt aggccagcca
    13021 tttttgagcg gccagcggcc gcgataggcc gacgcgaagc ggcggggcgt agggagcgca
    13081 gcgaccgaag ggtaggcgct ttttgcagct cttcggctgt gcgctggcca gacagttatg
    13141 cacaggccag gcgggtttta agagttttaa taagttttaa agagttttag gcggaaaaat
    13201 cgcctttttt ctcttttata tcagtcactt acatgtgtga ccggttccca atgtacggct
    13261 ttgggttccc aatgtacggg ttccggttcc caatgtacgg ctttgggttc ccaatgtacg
    13321 tgctatccac aggaaacaga ccttttcgac ctttttcccc tgctagggca atttgcccta
    13381 gcatctgctc cgtacattag gaaccggcgg atgcttcgcc ctcgatcagg ttgcggtagc
    13441 gcatgactag gatcgggcca gcctgccccg cctcctcctt caaatcgtac tccggcaggt
    13501 catttgaccc gatcagcttg cgcacggtga aacagaactt cttgaactct ccggcgctgc
    13561 cactgcgttc gtagatcgtc ttgaacaacc atctggcttc tgccttgcct gcggcgcggc
    13621 gtgccaggcg gtagagaaaa cggccgatgc cgggatcgat caaaaagtaa tcggggtgaa
    13681 ccgtcagcac gtccgggttc ttgccttctg tgatctcgcg gtacatccaa tcagctagct
    13741 cgatctcgat gtactccggc cgcccggttt cgctctttac gatcttgtag cggctaatca
    13801 aggcttcacc ctcggatacc gtcaccaggc ggccgttctt ggccttcttc gtacgctgca
    13861 tggcaacgtg cgtggtgttt aaccgaatgc aggtttctac caggtcgtct ttctgctttc
    13921 cgccatcggc tcgccggcag aacttgagta cgtccgcaac gtgtggacgg aacacgcggc
    13981 cgggcttgtc tcccttccct tcccggtatc ggttcatgga ttcggttaga tgggaaaccg
    14041 ccatcagtac caggtcgtaa tcccacacac tggccatgcc ggccggccct gcggaaacct
    14101 ctacgtgccc gtctggaagc tcgtagcgga tcacctcgcc agctcgtcgg tcacgcttcg
    14161 acagacggaa aacggccacg tccatgatgc tgcgactatc gcgggtgccc acgtcataga
    14221 gcatcggaac gaaaaaatct ggttgctcgt cgcccttggg cggcttccta atcgacggcg
    14281 caccggctgc cggcggttgc cgggattctt tgcggattcg atcagcggcc gcttgccacg
    14341 attcaccggg gcgtgcttct gcctcgatgc gttgccgctg ggcggcctgc gcggccttca
    14401 acttctccac caggtcatca cccagcgccg cgccgatttg taccgggccg gatggtttgc
    14461 gaccgctcac gccgattcct cgggcttggg ggttccagtg ccattgcagg gccggcaggc
    14521 aacccagccg cttacgcctg gccaaccgcc cgttcctcca cacatggggc attccacggc
    14581 gtcggtgcct ggttgttctt gattttccat gccgcctcct ttagccgcta aaattcatct
    14641 actcatttat tcatttgctc atttactctg gtagctgcgc gatgtattca gatagcagct
    14701 cggtaatggt cttgccttgg cgtaccgcgt acatcttcag cttggtgtga tcctccgccg
    14761 gcaactgaaa gttgacccgc ttcatggctg gcgtgtctgc caggctggcc aacgttgcag
    14821 ccttgctgct gcgtgcgctc ggacggccgg cacttagcgt gtttgtgctt ttgctcattt
    14881 tctctttacc tcattaactc aaatgagttt tgatttaatt tcagcggcca gcgcctggac
    14941 ctcgcgggca gcgtcgccct cgggttctga ttcaagaacg gttgtgccgg cggcggcagt
    15001 gcctgggtag ctcacgcgct gcgtgatacg ggactcaaga atgggcagct cgtacccggc
    15061 cagcgcctcg gcaacctcac cgccgatgcg cgtgcctttg atcgcccgcg acacgacaaa
    15121 ggccgcttgt agccttccat ccgtgacctc aatgcgctgc ttaaccagct ccaccaggtc
    15181 ggcggtggcc catatgtcgt aagggcttgg ctgcaccgga atcagcacga agtcggctgc
    15241 cttgatcgcg gacacagcca agtccgccgc ctggggcgct ccgtcgatca ctacgaagtc
    15301 gcgccggccg atggccttca cgtcgcggtc aatcgtcggg cggtcgatgc cgacaacggt
    15361 tagcggttga tcttcccgca cggccgccca atcgcgggca ctgccctggg gatcggaatc
    15421 gactaacaga acatcggccc cggcgagttg cagggcgcgg gctagatggg ttgcgatggt
    15481 cgtcttgcct gacccgcctt tctggttaag tacagcgata accttcatgc gttccccttg
    15541 cgtatttgtt tatttactca tcgcatcata tacgcagcga ccgcatgacg caagctgttt
    15601 tactcaaata cacatcacct ttttagacgg cggcgctcgg tttcttcagc ggccaagctg
    15661 gccggccagg ccgccagctt ggcatcagac aaaccggcca ggatttcatg cagccgcacg
    15721 gttgagacgt gcgcgggcgg ctcgaacacg tacccggccg cgatcatctc cgcctcgatc
    15781 tcttcggtaa tgaaaaacgg ttcgtcctgg ccgtcctggt gcggtttcat gcttgttcct
    15841 cttggcgttc attctcggcg gccgccaggg cgtcggcctc ggtcaatgcg tcctcacgga
    15901 aggcaccgcg ccgcctggcc tcggtgggcg tcacttcctc gctgcgctca agtgcgcggt
    15961 acagggtcga gcgatgcacg ccaagcagtg cagccgcctc tttcacggtg cggccttcct
    16021 ggtcgatcag ctcgcgggcg tgcgcgatct gtgccggggt gagggtaggg cgggggccaa
    16081 acttcacgcc tcgggccttg gcggcctcgc gcccgctccg ggtgcggtcg atgattaggg
    16141 aacgctcgaa ctcggcaatg ccggcgaaca cggtcaacac catgcggccg gccggcgtgg
    16201 tggtgtcggc ccacggctct gccaggctac gcaggcccgc gccggcctcc tggatgcgct
    16261 cggcaatgtc cagtaggtcg cgggtgctgc gggccaggcg gtctagcctg gtcactgtca
    16321 caacgtcgcc agggcgtagg tggtcaagca tcctggccag ctccgggcgg tcgcgcctgg
    16381 tgccggtgat cttctcggaa aacagcttgg tgcagccggc cgcgtgcagt tcggcccgtt
    16441 ggttggtcaa gtcctggtcg tcggtgctga cgcgggcata gcccagcagg ccagcggcgg
    16501 cgctcttgtt catggcgtaa tgtctccggt tctagtcgca agtatt
//
