LOCUS       sequence-313816-       16795 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Egg cell-specific promoter-controlled expression of 
            3×FLAG-NLS-zCas9-NLS. Contains gRNA scaffold for insertion of target
            sequence (U6-26 promoter). Selection of transgenic seeds by mTomato 
            fluoresce..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16795)
  AUTHORS   Angulo J, Astin CP, Bauer O , Blash KJ, Bowen NM, Chukwudinma NJ, 
            Dinofrio AS, Faletti DO, Ghulam AM, Gusinde-Duffy CM, Horace KJ, 
            Ingram AM, Isaack KE, Kiser RJ, Kobylanski JS, Long MR, Manning GA, 
            Morales JM, Nguyen KH, Pham RT, Phillips MH, Reel TW, Seo JE, Vo HD,
            Wukuson AM, Yeary KA, Zheng GY, Lukowitz W
  TITLE     Targeted mutagenesis of the Arabidopsis GROWTH-REGULATING FACTOR 
            (GRF) gene family suggests competition of multiplexed sgRNAs for 
            Cas9 apoprotein
  JOURNAL   bioRxiv
REFERENCE   2  (bases 1 to 16795)
  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: "bioRxiv"
COMMENT     Sequence Label: pTEE
FEATURES             Location/Qualifiers
     source          1..16795
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        1..76
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             1646..1711
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1718..1738
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1763..5863
                     /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             1763..5863
                     /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             5776..5784
                     /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             5864..5911
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7402..8100
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGL
                     VTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQ
                     ALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHH
                     LFLYGMDELYK"
     primer_bind     7800..7819
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     7994..8017
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    8365..8539
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    8617..8641
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             9066..9860
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      9947..10535
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10436..10455
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10689..10706
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    10721..10861
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(10787..10806)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     10947..10969
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      11205..11399
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(11465..12538)
                     /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(12967..13596)
                     /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    14897..14921
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     15109..15131
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     15123..15140
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     15124..15140
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             15853..16644
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
ORIGIN
        1 gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt
       61 ggcaccgagt cggtgctttt ttttgcaaaa ttttccagat cgatttcttc ttcctctgtt
      121 cttcggcgtt caatttctgg ggttttctct tcgttttctg taactgaaac ctaaaatttg
      181 acctaaaaaa aatctcaaat aatatgattc agtggttttg tacttttcag ttagttgagt
      241 tttgcagttc cgatgagata aaccaatacc atggttatac tagtgaataa aagcatttgc
      301 gtttggttta tcattgcgtt tatacaagga cagagatcca ctgagctgga atagcttaaa
      361 accattatca gaacaaaata aaccattttt tgttaagaat cagagcatag taaacaacag
      421 aaacaaccta agagaggtaa cttgtccaag aagatagcta attatatcta ttttataaaa
      481 gttatcatag tttgtaagtc acaaaagatg caaataacag agaaactagg agacttgaga
      541 atatacattc ttgtatattt gtattcgaga ttgtgaaaat ttgaccataa gtttaaattc
      601 ttaaaaagat atatctgatc taggtgatgg ttatagactg taattttacc acatgtttaa
      661 tgatggatag tgacacacat gacacatcga caacactata gcatcttatt tagattacaa
      721 catgaaattt ttctgtaata catgtctttg tacataattt aaaagtaatt cctaagaaat
      781 atatttatac aaggagttta aagaaaacat agcataaagt tcaatgagta gtaaaaacca
      841 tatacagtat atagcataaa gttcaatgag tttattacaa aagcattggt tcactttctg
      901 taacacgacg ttaaaccttc gtctccaata ggagcgctac tgattcaaca tgccaatata
      961 tactaaatac gtttctacag tcaaatgctt taacgtttca tgattaagtg actatttacc
     1021 gtcaatcctt tcccattcct cccactaatc caacttttta attactctta aatcaccact
     1081 aagctagtaa cgcctatcat gaattagctc tactaaatct agcaaccttt caaatttgca
     1141 gtattgcagg tgtctctgtg tctttaaaat agttgcctta tgatttcttc ggtttcaaga
     1201 tgatcaaata gttatagatt tcatgctcac acatgctcat tagatgtgta catactttac
     1261 ttacccaaat ctattttctc gcaaagattt tgatggtaaa gctgatttgg ttctattgaa
     1321 ctaaatcaaa cgagtttcag actgagtgat tctaatccgg cccattagcc cctaaacaga
     1381 cccactaatt acgcagcttt taatagagta attacaccta gtttacccac taaaccacta
     1441 agcactaatt atctcacaat ctaatgagct tccctcgtaa ttacttgggc tttcactcta
     1501 ccatttattt gtaacagtca agtctctact gtctctatat aaactctcta aagttaacac
     1561 acaattctca tcacaaacaa atcaaccaaa gcaacttcta ctctttcttc tttcgacctt
     1621 atcaatctgt tgagaaatct agatggatta caaggaccac gacggggatt acaaggacca
     1681 cgacattgat tacaaggatg atgatgacaa gatggctccg aagaagaaga ggaaggttgg
     1741 catccacggg gtgccagctg ctgacaagaa gtactcgatc ggcctcgata ttgggactaa
     1801 ctctgttggc tgggccgtga tcaccgacga gtacaaggtg ccctcaaaga agttcaaggt
     1861 cctgggcaac accgatcggc attccatcaa gaagaatctc attggcgctc tcctgttcga
     1921 cagcggcgag acggctgagg ctacgcggct caagcgcacc gcccgcaggc ggtacacgcg
     1981 caggaagaat cgcatctgct acctgcagga gattttctcc aacgagatgg cgaaggttga
     2041 cgattctttc ttccacaggc tggaggagtc attcctcgtg gaggaggata agaagcacga
     2101 gcggcatcca atcttcggca acattgtcga cgaggttgcc taccacgaga agtaccctac
     2161 gatctaccat ctgcggaaga agctcgtgga ctccacagat aaggcggacc tccgcctgat
     2221 ctacctcgct ctggcccaca tgattaagtt caggggccat ttcctgatcg agggggatct
     2281 caacccggac aatagcgatg ttgacaagct gttcatccag ctcgtgcaga cgtacaacca
     2341 gctcttcgag gagaacccca ttaatgcgtc aggcgtcgac gcgaaggcta tcctgtccgc
     2401 taggctctcg aagtctcggc gcctcgagaa cctgatcgcc cagctgccgg gcgagaagaa
     2461 gaacggcctg ttcgggaatc tcattgcgct cagcctgggg ctcacgccca acttcaagtc
     2521 gaatttcgat ctcgctgagg acgccaagct gcagctctcc aaggacacat acgacgatga
     2581 cctggataac ctcctggccc agatcggcga tcagtacgcg gacctgttcc tcgctgccaa
     2641 gaatctgtcg gacgccatcc tcctgtctga tattctcagg gtgaacaccg agattacgaa
     2701 ggctccgctc tcagcctcca tgatcaagcg ctacgacgag caccatcagg atctgaccct
     2761 cctgaaggcg ctggtcaggc agcagctccc cgagaagtac aaggagatct tcttcgatca
     2821 gtcgaagaac ggctacgctg ggtacattga cggcggggcc tctcaggagg agttctacaa
     2881 gttcatcaag ccgattctgg agaagatgga cggcacggag gagctgctgg tgaagctcaa
     2941 tcgcgaggac ctcctgagga agcagcggac attcgataac ggcagcatcc cacaccagat
     3001 tcatctcggg gagctgcacg ctatcctgag gaggcaggag gacttctacc ctttcctcaa
     3061 ggataaccgc gagaagatcg agaagattct gactttcagg atcccgtact acgtcggccc
     3121 actcgctagg ggcaactccc gcttcgcttg gatgacccgc aagtcagagg agacgatcac
     3181 gccgtggaac ttcgaggagg tggtcgacaa gggcgctagc gctcagtcgt tcatcgagag
     3241 gatgacgaat ttcgacaaga acctgccaaa tgagaaggtg ctccctaagc actcgctcct
     3301 gtacgagtac ttcacagtct acaacgagct gactaaggtg aagtatgtga ccgagggcat
     3361 gaggaagccg gctttcctgt ctggggagca gaagaaggcc atcgtggacc tcctgttcaa
     3421 gaccaaccgg aaggtcacgg ttaagcagct caaggaggac tacttcaaga agattgagtg
     3481 cttcgattcg gtcgagatct ctggcgttga ggaccgcttc aacgcctccc tggggaccta
     3541 ccacgatctc ctgaagatca ttaaggataa ggacttcctg gacaacgagg agaatgagga
     3601 tatcctcgag gacattgtgc tgacactcac tctgttcgag gaccgggaga tgatcgagga
     3661 gcgcctgaag acttacgccc atctcttcga tgacaaggtc atgaagcagc tcaagaggag
     3721 gaggtacacc ggctggggga ggctgagcag gaagctcatc aacggcattc gggacaagca
     3781 gtccgggaag acgatcctcg acttcctgaa gagcgatggc ttcgcgaacc gcaatttcat
     3841 gcagctgatt cacgatgaca gcctcacatt caaggaggat atccagaagg ctcaggtgag
     3901 cggccagggg gactcgctgc acgagcatat cgcgaacctc gctggctcgc cagctatcaa
     3961 gaaggggatt ctgcagaccg tgaaggttgt ggacgagctg gtgaaggtca tgggcaggca
     4021 caagcctgag aacatcgtca ttgagatggc ccgggagaat cagaccacgc agaagggcca
     4081 gaagaactca cgcgagagga tgaagaggat cgaggagggc attaaggagc tggggtccca
     4141 gatcctcaag gagcacccgg tggagaacac gcagctgcag aatgagaagc tctacctgta
     4201 ctacctccag aatggccgcg atatgtatgt ggaccaggag ctggatatta acaggctcag
     4261 cgattacgac gtcgatcata tcgttccaca gtcattcctg aaggatgact ccattgacaa
     4321 caaggtcctc accaggtcgg acaagaaccg gggcaagtct gataatgttc cttcagagga
     4381 ggtcgttaag aagatgaaga actactggcg ccagctcctg aatgccaagc tgatcacgca
     4441 gcggaagttc gataacctca caaaggctga gaggggcggg ctctctgagc tggacaaggc
     4501 gggcttcatc aagaggcagc tggtcgagac acggcagatc actaagcacg ttgcgcagat
     4561 tctcgactca cggatgaaca ctaagtacga tgagaatgac aagctgatcc gcgaggtgaa
     4621 ggtcatcacc ctgaagtcaa agctcgtctc cgacttcagg aaggatttcc agttctacaa
     4681 ggttcgggag atcaacaatt accaccatgc ccatgacgcg tacctgaacg cggtggtcgg
     4741 cacagctctg atcaagaagt acccaaagct cgagagcgag ttcgtgtacg gggactacaa
     4801 ggtttacgat gtgaggaaga tgatcgccaa gtcggagcag gagattggca aggctaccgc
     4861 caagtacttc ttctactcta acattatgaa tttcttcaag acagagatca ctctggccaa
     4921 tggcgagatc cggaagcgcc ccctcatcga gacgaacggc gagacggggg agatcgtgtg
     4981 ggacaagggc agggatttcg cgaccgtcag gaaggttctc tccatgccac aagtgaatat
     5041 cgtcaagaag acagaggtcc agactggcgg gttctctaag gagtcaattc tgcctaagcg
     5101 gaacagcgac aagctcatcg cccgcaagaa ggactgggat ccgaagaagt acggcgggtt
     5161 cgacagcccc actgtggcct actcggtcct ggttgtggcg aaggttgaga agggcaagtc
     5221 caagaagctc aagagcgtga aggagctgct ggggatcacg attatggagc gctccagctt
     5281 cgagaagaac ccgatcgatt tcctggaggc gaagggctac aaggaggtga agaaggacct
     5341 gatcattaag ctccccaagt actcactctt cgagctggag aacggcagga agcggatgct
     5401 ggcttccgct ggcgagctgc agaaggggaa cgagctggct ctgccgtcca agtatgtgaa
     5461 cttcctctac ctggcctccc actacgagaa gctcaagggc agccccgagg acaacgagca
     5521 gaagcagctg ttcgtcgagc agcacaagca ttacctcgac gagatcattg agcagatttc
     5581 cgagttctcc aagcgcgtga tcctggccga cgcgaatctg gataaggtcc tctccgcgta
     5641 caacaagcac cgcgacaagc caatcaggga gcaggctgag aatatcattc atctcttcac
     5701 cctgacgaac ctcggcgccc ctgctgcttt caagtacttc gacacaacta tcgatcgcaa
     5761 gaggtacaca agcactaagg aggtcctgga cgcgaccctc atccaccagt cgattaccgg
     5821 cctctacgag acgcgcatcg acctgtctca gctcgggggc gacaagcggc cagcggcgac
     5881 gaagaaggcg gggcaggcga agaagaagaa gtgagctcag agctttcgtt cgtatcatcg
     5941 gtttcgacaa cgttcgtcaa gttcaatgca tcagtttcat tgcgcacaca ccagaatcct
     6001 actgagtttg agtattatgg cattgggaaa actgtttttc ttgtaccatt tgttgtgctt
     6061 gtaatttact gtgtttttta ttcggttttc gctatcgaac tgtgaaatgg aaatggatgg
     6121 agaagagtta atgaatgata tggtcctttt gttcattctc aaattaatat tatttgtttt
     6181 ttctcttatt tgttgtgtgt tgaatttgaa attataagag atatgcaaac attttgtttt
     6241 gagtaaaaat gtgtcaaatc gtggcctcta atgaccgaag ttaatatgag gagtaaaaca
     6301 cttgtagttg taccattatg cttattcact aggcaacaaa tatattttca gacctagaaa
     6361 agctgcaaat gttactgaat acaagtatgt cctcttgtgt tttagacatt tatgaacttt
     6421 cctttatgta attttccaga atccttgtca gattctaatc attgctttat aattatagtt
     6481 atactcatgg atttgtagtt gagtatgaaa atatttttta atgcatttta tgacttgcca
     6541 attgattgac aacgaattcg ggcccattta aataggcgaa aggggtttag ggtttaagct
     6601 cagacttcta ttggagtaaa tgggacggtg tcacattttc cgttttggaa atgaactttg
     6661 ggctcacgtt atgggctatt agatatttga tgggctttct agtaaataca atataagtta
     6721 ttgggcttag tttaaataag cccatgttgg aaatatttga cacatgtctt ggctactagt
     6781 gctaaacatg caaccgaaca gttgtcgaga caagtcgcag catatacaat ggatcaaaca
     6841 cgcctagtgt cgccgcgtct cgctcatgtg tcaccttgtt tcctcgtttt tttttaattt
     6901 ttcataagtt cttttgtttt atcttcaata caaatttttg gctgtatctt gcaaactctt
     6961 cgatcatatc gccaatatac gtgaacactg gtgatctaat ttgttgtgtt aattgttaaa
     7021 tttagattct attctccggt ttaaaagtga attatatgta tcatggttaa aacattgtaa
     7081 gtaagatgat aataaaatga taaatttagt tgatggataa cgtgaagcaa aaaatgagat
     7141 agatacattt gattttgtcg tattttgaca tatgcggaga gtgagctacg cgcatgaaga
     7201 tcaagagaca cttgctcgag ctcacagagt gacgtgtaaa aagcttagac tgaagtcccc
     7261 atgcaaacct aatcctacgt ggctcaaacc acgagctcac ttgacaatat ataaactcct
     7321 cctaagtccc gttctcttca tccatctctc acaacaaaca aaaagaaaat ggttaagctc
     7381 agcaatctcc tcgttggatc tatgagcaag ggcgaggagg tcatcaaaga gttcatgcgc
     7441 ttcaaggtgc gcatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc
     7501 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggcggcccc
     7561 ctgcccttcg cctgggacat cctgtccccc cagttcatgt acggctccaa ggcgtacgtg
     7621 aagcaccccg ccgacatccc cgattacaag aagctgtcct tccccgaggg cttcaagtgg
     7681 gagcgcgtga tgaacttcga ggacggcggt ctggtgaccg tgacccagga ctcctccctg
     7741 caggacggca cgctgatcta caaggtgaag atgcgcggca ccaacttccc ccccgacggc
     7801 cccgtaatgc agaagaagac catgggctgg gaggcctcca ccgagcgcct gtacccccgc
     7861 gacggcgtgc tgaagggcga gatccaccag gccctgaagc tgaaggacgg cggccactac
     7921 ctggtggagt tcaagaccat ctacatggcc aagaagcccg tgcaactgcc cggctactac
     7981 tacgtggaca ccaagctgga catcacctcc cacaacgagg actacaccat cgtggaacag
     8041 tacgagcgct ccgagggccg ccaccacctg ttcctgtacg gcatggacga gctgtacaag
     8101 cttaagaaga agaaattcaa gcgtgaagag caagcaagga aagctaaggt gaacaatgag
     8161 aaaaagacgg aaatagtgaa accagagagt tgtagcaatg aaggagatgt caaggatctg
     8221 aaaagaaagg actctgagga tggaaacgag ggtgaggaag aagaagcttc ttcgaaaccg
     8281 aaaaagccaa aagttgctct ttctcatctt caggacattg acgacacaga agctgatcaa
     8341 gaagaagagt aatctagtct cgagtttctc cataataatg tgtgagtagt tcccagataa
     8401 gggaattagg gttcctatag ggtttcgctc atgtgttgag catataagaa acccttagta
     8461 tgtatttgta tttgtaaaat acttctatca ataaaatttc taattcctaa aaccaaaatc
     8521 cagtactaaa atccagatcc cccgaattaa ttcggcgtta attcagtaca ttaaaaacgt
     8581 ccgcaatgtg ttattaagtt gtctaagcgt caatttgttt acaccacaat atatcctgcc
     8641 accagccagc caacagctcc ccgaccggca gctcggcaca aaatcaccac tcgatacagg
     8701 cagcccatca gtccgggacg gcgtcagcgg gagagccgtt gtaaggcggc agactttgct
     8761 catgttaccg atgctattcg gaagaacggc aactaagctg ccgggtttga aacacggatg
     8821 atctcgcgga gggtagcatg ttgattgtaa cgatgacaga gcgttgctgc ctgtgatcac
     8881 cgcggtttca aaatcggctc cgtcgatact atgttatacg ccaactttga aaacaacttt
     8941 gaaaaagctg ttttctggta tttaaggttt tagaatgcaa ggaacagtga attggagttc
     9001 gtcttgttat aattagcttc ttggggtatc tttaaatact gtagaaaaga ggaaggaaat
     9061 aataaatggc taaaatgaga atatcaccgg aattgaaaaa actgatcgaa aaataccgct
     9121 gcgtaaaaga tacggaagga atgtctcctg ctaaggtata taagctggtg ggagaaaatg
     9181 aaaacctata tttaaaaatg acggacagcc ggtataaagg gaccacctat gatgtggaac
     9241 gggaaaagga catgatgcta tggctggaag gaaagctgcc tgttccaaag gtcctgcact
     9301 ttgaacggca tgatggctgg agcaatctgc tcatgagtga ggccgatggc gtcctttgct
     9361 cggaagagta tgaagatgaa caaagccctg aaaagattat cgagctgtat gcggagtgca
     9421 tcaggctctt tcactccatc gacatatcgg attgtcccta tacgaatagc ttagacagcc
     9481 gcttagccga attggattac ttactgaata acgatctggc cgatgtggat tgcgaaaact
     9541 gggaagaaga cactccattt aaagatccgc gcgagctgta tgatttttta aagacggaaa
     9601 agcccgaaga ggaacttgtc ttttcccacg gcgacctggg agacagcaac atctttgtga
     9661 aagatggcaa agtaagtggc tttattgatc ttgggagaag cggcagggcg gacaagtggt
     9721 atgacattgc cttctgcgtc cggtcgatca gggaggatat cggggaagaa cagtatgtcg
     9781 agctattttt tgacttactg gggatcaagc ctgattggga gaaaataaaa tattatattt
     9841 tactggatga attgttttag tacctagaat gcatgaccaa aatcccttaa cgtgagtttt
     9901 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     9961 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
    10021 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
    10081 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
    10141 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
    10201 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
    10261 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
    10321 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
    10381 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
    10441 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
    10501 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
    10561 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
    10621 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
    10681 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
    10741 ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg
    10801 atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc
    10861 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
    10921 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
    10981 atcaccgaaa cgcgcgaggc agggtgcctt gatgtgggcg ccggcggtcg agtggcgacg
    11041 gcgcggcttg tccgcgccct ggtagattgc ctggccgtag gccagccatt tttgagcggc
    11101 cagcggccgc gataggccga cgcgaagcgg cggggcgtag ggagcgcagc gaccgaaggg
    11161 taggcgcttt ttgcagctct tcggctgtgc gctggccaga cagttatgca caggccaggc
    11221 gggttttaag agttttaata agttttaaag agttttaggc ggaaaaatcg ccttttttct
    11281 cttttatatc agtcacttac atgtgtgacc ggttcccaat gtacggcttt gggttcccaa
    11341 tgtacgggtt ccggttccca atgtacggct ttgggttccc aatgtacgtg ctatccacag
    11401 gaaacagacc ttttcgacct ttttcccctg ctagggcaat ttgccctagc atctgctccg
    11461 tacattagga accggcggat gcttcgccct cgatcaggtt gcggtagcgc atgactagga
    11521 tcgggccagc ctgccccgcc tcctccttca aatcgtactc cggcaggtca tttgacccga
    11581 tcagcttgcg cacggtgaaa cagaacttct tgaactctcc ggcgctgcca ctgcgttcgt
    11641 agatcgtctt gaacaaccat ctggcttctg ccttgcctgc ggcgcggcgt gccaggcggt
    11701 agagaaaacg gccgatgccg ggatcgatca aaaagtaatc ggggtgaacc gtcagcacgt
    11761 ccgggttctt gccttctgtg atctcgcggt acatccaatc agctagctcg atctcgatgt
    11821 actccggccg cccggtttcg ctctttacga tcttgtagcg gctaatcaag gcttcaccct
    11881 cggataccgt caccaggcgg ccgttcttgg ccttcttcgt acgctgcatg gcaacgtgcg
    11941 tggtgtttaa ccgaatgcag gtttctacca ggtcgtcttt ctgctttccg ccatcggctc
    12001 gccggcagaa cttgagtacg tccgcaacgt gtggacggaa cacgcggccg ggcttgtctc
    12061 ccttcccttc ccggtatcgg ttcatggatt cggttagatg ggaaaccgcc atcagtacca
    12121 ggtcgtaatc ccacacactg gccatgccgg ccggccctgc ggaaacctct acgtgcccgt
    12181 ctggaagctc gtagcggatc acctcgccag ctcgtcggtc acgcttcgac agacggaaaa
    12241 cggccacgtc catgatgctg cgactatcgc gggtgcccac gtcatagagc atcggaacga
    12301 aaaaatctgg ttgctcgtcg cccttgggcg gcttcctaat cgacggcgca ccggctgccg
    12361 gcggttgccg ggattctttg cggattcgat cagcggccgc ttgccacgat tcaccggggc
    12421 gtgcttctgc ctcgatgcgt tgccgctggg cggcctgcgc ggccttcaac ttctccacca
    12481 ggtcatcacc cagcgccgcg ccgatttgta ccgggccgga tggtttgcga ccgctcacgc
    12541 cgattcctcg ggcttggggg ttccagtgcc attgcagggc cggcaggcaa cccagccgct
    12601 tacgcctggc caaccgcccg ttcctccaca catggggcat tccacggcgt cggtgcctgg
    12661 ttgttcttga ttttccatgc cgcctccttt agccgctaaa attcatctac tcatttattc
    12721 atttgctcat ttactctggt agctgcgcga tgtattcaga tagcagctcg gtaatggtct
    12781 tgccttggcg taccgcgtac atcttcagct tggtgtgatc ctccgccggc aactgaaagt
    12841 tgacccgctt catggctggc gtgtctgcca ggctggccaa cgttgcagcc ttgctgctgc
    12901 gtgcgctcgg acggccggca cttagcgtgt ttgtgctttt gctcattttc tctttacctc
    12961 attaactcaa atgagttttg atttaatttc agcggccagc gcctggacct cgcgggcagc
    13021 gtcgccctcg ggttctgatt caagaacggt tgtgccggcg gcggcagtgc ctgggtagct
    13081 cacgcgctgc gtgatacggg actcaagaat gggcagctcg tacccggcca gcgcctcggc
    13141 aacctcaccg ccgatgcgcg tgcctttgat cgcccgcgac acgacaaagg ccgcttgtag
    13201 ccttccatcc gtgacctcaa tgcgctgctt aaccagctcc accaggtcgg cggtggccca
    13261 tatgtcgtaa gggcttggct gcaccggaat cagcacgaag tcggctgcct tgatcgcgga
    13321 cacagccaag tccgccgcct ggggcgctcc gtcgatcact acgaagtcgc gccggccgat
    13381 ggccttcacg tcgcggtcaa tcgtcgggcg gtcgatgccg acaacggtta gcggttgatc
    13441 ttcccgcacg gccgcccaat cgcgggcact gccctgggga tcggaatcga ctaacagaac
    13501 atcggccccg gcgagttgca gggcgcgggc tagatgggtt gcgatggtcg tcttgcctga
    13561 cccgcctttc tggttaagta cagcgataac cttcatgcgt tccccttgcg tatttgttta
    13621 tttactcatc gcatcatata cgcagcgacc gcatgacgca agctgtttta ctcaaataca
    13681 catcaccttt ttagacggcg gcgctcggtt tcttcagcgg ccaagctggc cggccaggcc
    13741 gccagcttgg catcagacaa accggccagg atttcatgca gccgcacggt tgagacgtgc
    13801 gcgggcggct cgaacacgta cccggccgcg atcatctccg cctcgatctc ttcggtaatg
    13861 aaaaacggtt cgtcctggcc gtcctggtgc ggtttcatgc ttgttcctct tggcgttcat
    13921 tctcggcggc cgccagggcg tcggcctcgg tcaatgcgtc ctcacggaag gcaccgcgcc
    13981 gcctggcctc ggtgggcgtc acttcctcgc tgcgctcaag tgcgcggtac agggtcgagc
    14041 gatgcacgcc aagcagtgca gccgcctctt tcacggtgcg gccttcctgg tcgatcagct
    14101 cgcgggcgtg cgcgatctgt gccggggtga gggtagggcg ggggccaaac ttcacgcctc
    14161 gggccttggc ggcctcgcgc ccgctccggg tgcggtcgat gattagggaa cgctcgaact
    14221 cggcaatgcc ggcgaacacg gtcaacacca tgcggccggc cggcgtggtg gtgtcggccc
    14281 acggctctgc caggctacgc aggcccgcgc cggcctcctg gatgcgctcg gcaatgtcca
    14341 gtaggtcgcg ggtgctgcgg gccaggcggt ctagcctggt cactgtcaca acgtcgccag
    14401 ggcgtaggtg gtcaagcatc ctggccagct ccgggcggtc gcgcctggtg ccggtgatct
    14461 tctcggaaaa cagcttggtg cagccggccg cgtgcagttc ggcccgttgg ttggtcaagt
    14521 cctggtcgtc ggtgctgacg cgggcatagc ccagcaggcc agcggcggcg ctcttgttca
    14581 tggcgtaatg tctccggttc tagtcgcaag tattctactt tatgcgacta aaacacgcga
    14641 caagaaaacg ccaggaaaag ggcagggcgg cagcctgtcg cgtaacttag gacttgtgcg
    14701 acatgtcgtt ttcagaagac ggctgcactg aacgtcagaa gccgactgca ctatagcagc
    14761 ggaggggttg gatcaaagta ctttgatccc gaggggaacc ctgtggttgg catgcacata
    14821 caaatggacg aacggataaa ccttttcacg cccttttaaa tatccgatta ttctaataaa
    14881 cgctcttttc tcttaggttt acccgccaat atatcctgtc aaacactgat agtttaaact
    14941 gaaggcggga aacgacaatc tgatccaagc tcaagctgct ctagcattcg ccattcaggc
    15001 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
    15061 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
    15121 gttgtaaaac gacggccagt gccaagcttc gacttgcctt ccgcacaata catcatttct
    15181 tcttagcttt ttttcttctt cttcgttcat acagtttttt tttgtttatc agcttacatt
    15241 ttcttgaacc gtagctttcg ttttcttctt tttaactttc cattcggagt ttttgtatct
    15301 tgtttcatag tttgtcccag gattagaatg attaggcatc gaaccttcaa gaatttgatt
    15361 gaataaaaca tcttcattct taagatatga agataatctt caaaaggccc ctgggaatct
    15421 gaaagaagag aagcaggccc atttatatgg gaaagaacaa tagtatttct tatataggcc
    15481 catttaagtt gaaaacaatc ttcaaaagtc ccacatcgct tagataagaa aacgaagctg
    15541 agtttatata cagctagagt cgaagtagtg attgggagac caacccagtg gacataagcc
    15601 tgttcggttc gtaagctgta atgcaagtag cgtatgcgct cacgcaactg gtccagaacc
    15661 ttgaccgaac gcagcggtgg taacggcgca gtggcggttt tcatggcttg ttatgactgt
    15721 ttttttgggg tacagtctat gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt
    15781 cgatgtttga tgttatggag cagcaacgat gttacgcagc agggcagtcg ccctaaaaca
    15841 aagttaaaca tcatggggga agcggtgatc gccgaagtat cgactcaact atcagaggta
    15901 gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt gtacggctcc
    15961 gcagtggatg gcggcctgaa gccacacagt gatattgatt tgctggttac ggtgaccgta
    16021 aggcttgatg aaacaacgcg gcgagctttg atcaacgacc ttttggaaac ttcggcttcc
    16081 cctggagaga gcgagattct ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc
    16141 attccgtggc gttatccagc taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac
    16201 attcttgcag gtatcttcga gccagccacg atcgacattg atctggctat cttgctgaca
    16261 aaagcaagag aacatagcgt tgccttggta ggtccagcgg cggaggaact ctttgatccg
    16321 gttcctgaac aggatctatt tgaggcgcta aatgaaacct taacgctatg gaactcgccg
    16381 cccgactggg ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc
    16441 gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat ggagcgcctg
    16501 ccggcccagt atcagcccgt catacttgaa gctagacagg cttatcttgg acaagaagaa
    16561 gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg tccactacgt gaaaggcgag
    16621 atcaccaagg tagtcggcaa ataatgtcta gctagaaatt cgttcaagcc gacgccgctt
    16681 cgcggcgcgg cttaactcaa gcgttagatg cactaagcac ataattgctc acagccaaac
    16741 tatcaggtca agtctgcttt tattattttt aagcgtgcat aataagccgg tctcg
//
