LOCUS       sequence-313818-       17426 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ubiquitin 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 17426)
  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 17426)
  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: pTUU
FEATURES             Location/Qualifiers
     source          1..17426
                     /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             2277..2342
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2349..2369
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2394..6494
                     /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             2394..6494
                     /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             6407..6415
                     /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             6495..6542
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8033..8731
                     /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     8431..8450
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     8625..8648
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    8996..9170
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    9248..9272
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             9697..10491
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      10578..11166
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11067..11086
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11320..11337
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    11352..11492
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(11418..11437)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     11578..11600
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      11836..12030
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(12096..13169)
                     /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(13598..14227)
                     /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    15528..15552
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     15740..15762
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     15754..15771
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     15755..15771
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             16484..17275
                     /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 atggatacac agtctagctc aacagagctt
      301 ttaacccaaa ttggtacaat agaatacaac tttagatcat aattctcaaa agaaagagat
      361 tccttagcta ttctatctgc cactccattt ccttctcggc ttgtatgcac aagcataaaa
      421 tcctcaaact tgctaagtag atactttatg tcttggataa ttggattgag acttgacaag
      481 cataactttc atgtaaccaa agacacaagt tgctgagaat ccacctcaaa aatgatcttc
      541 ctataattga atcgggataa tgacagcaca gcccatctaa gagcctccac ttctacttcc
      601 agcacgcttc ttacttttac cacagctctt gcacctaacc ataacacctt ccctgtatga
      661 tcgcgaagca cccaccctaa gccacatttt aatccttctg ttggccatgc cccatcaaag
      721 ttgcacttaa cccaagattg tggtggagct tcccatgttt ctcgtctgtc ccgacggtgt
      781 tgtggttggt gctttcctta cattctgagc ctctttcctt ctaatccact catctgcatc
      841 ttcttgtgtc cttactaata cctcattggt tccaaattcc ctccctttaa gcaccagctc
      901 gtttctgttc ttccacagcc tcccaagtat ccaagggact aaagcctcca cattcttcag
      961 atcaggatat tcttgtttaa gatgttgaac tctatggagg tttgtatgaa ctgatgatct
     1021 aggaccggat aagttccctt cttcatagcg aacttattca aagaatgttt tgtgtatcat
     1081 tcttgttaca ttgttattaa tgaaaaaata ttattggtca ttggactgaa cacgagtgtt
     1141 aaatatggac caggccccaa ataagatcca ttgatatatg aattaaataa caagaataaa
     1201 tcgagtcacc aaaccacttg ccttttttaa cgagacttgt tcaccaactt gatacaaaag
     1261 tcattatcct atgcaaatca ataatcatac aaaaatatcc aataacacta aaaaattaaa
     1321 agaaatggat aatttcacaa tatgttatac gataaagaag ttacttttcc aagaaattca
     1381 ctgattttat aagcccactt gcattagata aatggcaaaa aaaaacaaaa aggaaaagaa
     1441 ataaagcacg aagaattcta gaaaatacga aatacgcttc aatgcagtgg gacccacggt
     1501 tcaattattg ccaattttca gctccaccgt atatttaaaa aataaaacga taatgctaaa
     1561 aaaatataaa tcgtaacgat cgttaaatct caacggctgg atcttatgac gaccgttaga
     1621 aattgtggtt gtcgacgagt cagtaataaa cggcgtcaaa gtggttgcag ccggcacaca
     1681 cgagtcgtgt ttatcaactc aaagcacaaa tacttttcct caacctaaaa ataaggcaat
     1741 tagccaaaaa caactttgcg tgtaaacaac gctcaataca cgtgtcattt tattattagc
     1801 tattgcttca ccgccttagc tttctcgtga cctagtcgtc ctcgtctttt cttcttcttc
     1861 ttctataaaa caatacccaa agagctcttc ttcttcacaa ttcagatttc aatttctcaa
     1921 aatcttaaaa actttctctc aattctctct accgtgatca aggtaaattt ctgtgttcct
     1981 tattctctca aaatcttcga ttttgttttc gttcgatccc aatttcgtat atgttctttg
     2041 gtttagattc tgttaatctt agatcgaaga cgattttctg ggtttgatcg ttagatatca
     2101 tcttaattct cgattagggt ttcatagata tcatccgatt tgttcaaata atttgagttt
     2161 tgtcgaataa ttactcttcg atttgtgatt tctatctaga tctggtgtta gtttctagtt
     2221 tgtgcgatcg aatttgtcga ttaatctgag tttttctgat taacaggttc tagatggatt
     2281 acaaggacca cgacggggat tacaaggacc acgacattga ttacaaggat gatgatgaca
     2341 agatggctcc gaagaagaag aggaaggttg gcatccacgg ggtgccagct gctgacaaga
     2401 agtactcgat cggcctcgat attgggacta actctgttgg ctgggccgtg atcaccgacg
     2461 agtacaaggt gccctcaaag aagttcaagg tcctgggcaa caccgatcgg cattccatca
     2521 agaagaatct cattggcgct ctcctgttcg acagcggcga gacggctgag gctacgcggc
     2581 tcaagcgcac cgcccgcagg cggtacacgc gcaggaagaa tcgcatctgc tacctgcagg
     2641 agattttctc caacgagatg gcgaaggttg acgattcttt cttccacagg ctggaggagt
     2701 cattcctcgt ggaggaggat aagaagcacg agcggcatcc aatcttcggc aacattgtcg
     2761 acgaggttgc ctaccacgag aagtacccta cgatctacca tctgcggaag aagctcgtgg
     2821 actccacaga taaggcggac ctccgcctga tctacctcgc tctggcccac atgattaagt
     2881 tcaggggcca tttcctgatc gagggggatc tcaacccgga caatagcgat gttgacaagc
     2941 tgttcatcca gctcgtgcag acgtacaacc agctcttcga ggagaacccc attaatgcgt
     3001 caggcgtcga cgcgaaggct atcctgtccg ctaggctctc gaagtctcgg cgcctcgaga
     3061 acctgatcgc ccagctgccg ggcgagaaga agaacggcct gttcgggaat ctcattgcgc
     3121 tcagcctggg gctcacgccc aacttcaagt cgaatttcga tctcgctgag gacgccaagc
     3181 tgcagctctc caaggacaca tacgacgatg acctggataa cctcctggcc cagatcggcg
     3241 atcagtacgc ggacctgttc ctcgctgcca agaatctgtc ggacgccatc ctcctgtctg
     3301 atattctcag ggtgaacacc gagattacga aggctccgct ctcagcctcc atgatcaagc
     3361 gctacgacga gcaccatcag gatctgaccc tcctgaaggc gctggtcagg cagcagctcc
     3421 ccgagaagta caaggagatc ttcttcgatc agtcgaagaa cggctacgct gggtacattg
     3481 acggcggggc ctctcaggag gagttctaca agttcatcaa gccgattctg gagaagatgg
     3541 acggcacgga ggagctgctg gtgaagctca atcgcgagga cctcctgagg aagcagcgga
     3601 cattcgataa cggcagcatc ccacaccaga ttcatctcgg ggagctgcac gctatcctga
     3661 ggaggcagga ggacttctac cctttcctca aggataaccg cgagaagatc gagaagattc
     3721 tgactttcag gatcccgtac tacgtcggcc cactcgctag gggcaactcc cgcttcgctt
     3781 ggatgacccg caagtcagag gagacgatca cgccgtggaa cttcgaggag gtggtcgaca
     3841 agggcgctag cgctcagtcg ttcatcgaga ggatgacgaa tttcgacaag aacctgccaa
     3901 atgagaaggt gctccctaag cactcgctcc tgtacgagta cttcacagtc tacaacgagc
     3961 tgactaaggt gaagtatgtg accgagggca tgaggaagcc ggctttcctg tctggggagc
     4021 agaagaaggc catcgtggac ctcctgttca agaccaaccg gaaggtcacg gttaagcagc
     4081 tcaaggagga ctacttcaag aagattgagt gcttcgattc ggtcgagatc tctggcgttg
     4141 aggaccgctt caacgcctcc ctggggacct accacgatct cctgaagatc attaaggata
     4201 aggacttcct ggacaacgag gagaatgagg atatcctcga ggacattgtg ctgacactca
     4261 ctctgttcga ggaccgggag atgatcgagg agcgcctgaa gacttacgcc catctcttcg
     4321 atgacaaggt catgaagcag ctcaagagga ggaggtacac cggctggggg aggctgagca
     4381 ggaagctcat caacggcatt cgggacaagc agtccgggaa gacgatcctc gacttcctga
     4441 agagcgatgg cttcgcgaac cgcaatttca tgcagctgat tcacgatgac agcctcacat
     4501 tcaaggagga tatccagaag gctcaggtga gcggccaggg ggactcgctg cacgagcata
     4561 tcgcgaacct cgctggctcg ccagctatca agaaggggat tctgcagacc gtgaaggttg
     4621 tggacgagct ggtgaaggtc atgggcaggc acaagcctga gaacatcgtc attgagatgg
     4681 cccgggagaa tcagaccacg cagaagggcc agaagaactc acgcgagagg atgaagagga
     4741 tcgaggaggg cattaaggag ctggggtccc agatcctcaa ggagcacccg gtggagaaca
     4801 cgcagctgca gaatgagaag ctctacctgt actacctcca gaatggccgc gatatgtatg
     4861 tggaccagga gctggatatt aacaggctca gcgattacga cgtcgatcat atcgttccac
     4921 agtcattcct gaaggatgac tccattgaca acaaggtcct caccaggtcg gacaagaacc
     4981 ggggcaagtc tgataatgtt ccttcagagg aggtcgttaa gaagatgaag aactactggc
     5041 gccagctcct gaatgccaag ctgatcacgc agcggaagtt cgataacctc acaaaggctg
     5101 agaggggcgg gctctctgag ctggacaagg cgggcttcat caagaggcag ctggtcgaga
     5161 cacggcagat cactaagcac gttgcgcaga ttctcgactc acggatgaac actaagtacg
     5221 atgagaatga caagctgatc cgcgaggtga aggtcatcac cctgaagtca aagctcgtct
     5281 ccgacttcag gaaggatttc cagttctaca aggttcggga gatcaacaat taccaccatg
     5341 cccatgacgc gtacctgaac gcggtggtcg gcacagctct gatcaagaag tacccaaagc
     5401 tcgagagcga gttcgtgtac ggggactaca aggtttacga tgtgaggaag atgatcgcca
     5461 agtcggagca ggagattggc aaggctaccg ccaagtactt cttctactct aacattatga
     5521 atttcttcaa gacagagatc actctggcca atggcgagat ccggaagcgc cccctcatcg
     5581 agacgaacgg cgagacgggg gagatcgtgt gggacaaggg cagggatttc gcgaccgtca
     5641 ggaaggttct ctccatgcca caagtgaata tcgtcaagaa gacagaggtc cagactggcg
     5701 ggttctctaa ggagtcaatt ctgcctaagc ggaacagcga caagctcatc gcccgcaaga
     5761 aggactggga tccgaagaag tacggcgggt tcgacagccc cactgtggcc tactcggtcc
     5821 tggttgtggc gaaggttgag aagggcaagt ccaagaagct caagagcgtg aaggagctgc
     5881 tggggatcac gattatggag cgctccagct tcgagaagaa cccgatcgat ttcctggagg
     5941 cgaagggcta caaggaggtg aagaaggacc tgatcattaa gctccccaag tactcactct
     6001 tcgagctgga gaacggcagg aagcggatgc tggcttccgc tggcgagctg cagaagggga
     6061 acgagctggc tctgccgtcc aagtatgtga acttcctcta cctggcctcc cactacgaga
     6121 agctcaaggg cagccccgag gacaacgagc agaagcagct gttcgtcgag cagcacaagc
     6181 attacctcga cgagatcatt gagcagattt ccgagttctc caagcgcgtg atcctggccg
     6241 acgcgaatct ggataaggtc ctctccgcgt acaacaagca ccgcgacaag ccaatcaggg
     6301 agcaggctga gaatatcatt catctcttca ccctgacgaa cctcggcgcc cctgctgctt
     6361 tcaagtactt cgacacaact atcgatcgca agaggtacac aagcactaag gaggtcctgg
     6421 acgcgaccct catccaccag tcgattaccg gcctctacga gacgcgcatc gacctgtctc
     6481 agctcggggg cgacaagcgg ccagcggcga cgaagaaggc ggggcaggcg aagaagaaga
     6541 agtgagctca gagctttcgt tcgtatcatc ggtttcgaca acgttcgtca agttcaatgc
     6601 atcagtttca ttgcgcacac accagaatcc tactgagttt gagtattatg gcattgggaa
     6661 aactgttttt cttgtaccat ttgttgtgct tgtaatttac tgtgtttttt attcggtttt
     6721 cgctatcgaa ctgtgaaatg gaaatggatg gagaagagtt aatgaatgat atggtccttt
     6781 tgttcattct caaattaata ttatttgttt tttctcttat ttgttgtgtg ttgaatttga
     6841 aattataaga gatatgcaaa cattttgttt tgagtaaaaa tgtgtcaaat cgtggcctct
     6901 aatgaccgaa gttaatatga ggagtaaaac acttgtagtt gtaccattat gcttattcac
     6961 taggcaacaa atatattttc agacctagaa aagctgcaaa tgttactgaa tacaagtatg
     7021 tcctcttgtg ttttagacat ttatgaactt tcctttatgt aattttccag aatccttgtc
     7081 agattctaat cattgcttta taattatagt tatactcatg gatttgtagt tgagtatgaa
     7141 aatatttttt aatgcatttt atgacttgcc aattgattga caacgaattc gggcccattt
     7201 aaataggcga aaggggttta gggtttaagc tcagacttct attggagtaa atgggacggt
     7261 gtcacatttt ccgttttgga aatgaacttt gggctcacgt tatgggctat tagatatttg
     7321 atgggctttc tagtaaatac aatataagtt attgggctta gtttaaataa gcccatgttg
     7381 gaaatatttg acacatgtct tggctactag tgctaaacat gcaaccgaac agttgtcgag
     7441 acaagtcgca gcatatacaa tggatcaaac acgcctagtg tcgccgcgtc tcgctcatgt
     7501 gtcaccttgt ttcctcgttt ttttttaatt tttcataagt tcttttgttt tatcttcaat
     7561 acaaattttt ggctgtatct tgcaaactct tcgatcatat cgccaatata cgtgaacact
     7621 ggtgatctaa tttgttgtgt taattgttaa atttagattc tattctccgg tttaaaagtg
     7681 aattatatgt atcatggtta aaacattgta agtaagatga taataaaatg ataaatttag
     7741 ttgatggata acgtgaagca aaaaatgaga tagatacatt tgattttgtc gtattttgac
     7801 atatgcggag agtgagctac gcgcatgaag atcaagagac acttgctcga gctcacagag
     7861 tgacgtgtaa aaagcttaga ctgaagtccc catgcaaacc taatcctacg tggctcaaac
     7921 cacgagctca cttgacaata tataaactcc tcctaagtcc cgttctcttc atccatctct
     7981 cacaacaaac aaaaagaaaa tggttaagct cagcaatctc ctcgttggat ctatgagcaa
     8041 gggcgaggag gtcatcaaag agttcatgcg cttcaaggtg cgcatggagg gctccatgaa
     8101 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac
     8161 cgccaagctg aaggtgacca agggcggccc cctgcccttc gcctgggaca tcctgtcccc
     8221 ccagttcatg tacggctcca aggcgtacgt gaagcacccc gccgacatcc ccgattacaa
     8281 gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
     8341 tctggtgacc gtgacccagg actcctccct gcaggacggc acgctgatct acaaggtgaa
     8401 gatgcgcggc accaacttcc cccccgacgg ccccgtaatg cagaagaaga ccatgggctg
     8461 ggaggcctcc accgagcgcc tgtacccccg cgacggcgtg ctgaagggcg agatccacca
     8521 ggccctgaag ctgaaggacg gcggccacta cctggtggag ttcaagacca tctacatggc
     8581 caagaagccc gtgcaactgc ccggctacta ctacgtggac accaagctgg acatcacctc
     8641 ccacaacgag gactacacca tcgtggaaca gtacgagcgc tccgagggcc gccaccacct
     8701 gttcctgtac ggcatggacg agctgtacaa gcttaagaag aagaaattca agcgtgaaga
     8761 gcaagcaagg aaagctaagg tgaacaatga gaaaaagacg gaaatagtga aaccagagag
     8821 ttgtagcaat gaaggagatg tcaaggatct gaaaagaaag gactctgagg atggaaacga
     8881 gggtgaggaa gaagaagctt cttcgaaacc gaaaaagcca aaagttgctc tttctcatct
     8941 tcaggacatt gacgacacag aagctgatca agaagaagag taatctagtc tcgagtttct
     9001 ccataataat gtgtgagtag ttcccagata agggaattag ggttcctata gggtttcgct
     9061 catgtgttga gcatataaga aacccttagt atgtatttgt atttgtaaaa tacttctatc
     9121 aataaaattt ctaattccta aaaccaaaat ccagtactaa aatccagatc ccccgaatta
     9181 attcggcgtt aattcagtac attaaaaacg tccgcaatgt gttattaagt tgtctaagcg
     9241 tcaatttgtt tacaccacaa tatatcctgc caccagccag ccaacagctc cccgaccggc
     9301 agctcggcac aaaatcacca ctcgatacag gcagcccatc agtccgggac ggcgtcagcg
     9361 ggagagccgt tgtaaggcgg cagactttgc tcatgttacc gatgctattc ggaagaacgg
     9421 caactaagct gccgggtttg aaacacggat gatctcgcgg agggtagcat gttgattgta
     9481 acgatgacag agcgttgctg cctgtgatca ccgcggtttc aaaatcggct ccgtcgatac
     9541 tatgttatac gccaactttg aaaacaactt tgaaaaagct gttttctggt atttaaggtt
     9601 ttagaatgca aggaacagtg aattggagtt cgtcttgtta taattagctt cttggggtat
     9661 ctttaaatac tgtagaaaag aggaaggaaa taataaatgg ctaaaatgag aatatcaccg
     9721 gaattgaaaa aactgatcga aaaataccgc tgcgtaaaag atacggaagg aatgtctcct
     9781 gctaaggtat ataagctggt gggagaaaat gaaaacctat atttaaaaat gacggacagc
     9841 cggtataaag ggaccaccta tgatgtggaa cgggaaaagg acatgatgct atggctggaa
     9901 ggaaagctgc ctgttccaaa ggtcctgcac tttgaacggc atgatggctg gagcaatctg
     9961 ctcatgagtg aggccgatgg cgtcctttgc tcggaagagt atgaagatga acaaagccct
    10021 gaaaagatta tcgagctgta tgcggagtgc atcaggctct ttcactccat cgacatatcg
    10081 gattgtccct atacgaatag cttagacagc cgcttagccg aattggatta cttactgaat
    10141 aacgatctgg ccgatgtgga ttgcgaaaac tgggaagaag acactccatt taaagatccg
    10201 cgcgagctgt atgatttttt aaagacggaa aagcccgaag aggaacttgt cttttcccac
    10261 ggcgacctgg gagacagcaa catctttgtg aaagatggca aagtaagtgg ctttattgat
    10321 cttgggagaa gcggcagggc ggacaagtgg tatgacattg ccttctgcgt ccggtcgatc
    10381 agggaggata tcggggaaga acagtatgtc gagctatttt ttgacttact ggggatcaag
    10441 cctgattggg agaaaataaa atattatatt ttactggatg aattgtttta gtacctagaa
    10501 tgcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
    10561 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
    10621 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
    10681 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
    10741 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
    10801 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
    10861 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
    10921 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
    10981 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
    11041 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
    11101 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
    11161 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
    11221 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
    11281 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
    11341 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
    11401 atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
    11461 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
    11521 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
    11581 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagggtgcct
    11641 tgatgtgggc gccggcggtc gagtggcgac ggcgcggctt gtccgcgccc tggtagattg
    11701 cctggccgta ggccagccat ttttgagcgg ccagcggccg cgataggccg acgcgaagcg
    11761 gcggggcgta gggagcgcag cgaccgaagg gtaggcgctt tttgcagctc ttcggctgtg
    11821 cgctggccag acagttatgc acaggccagg cgggttttaa gagttttaat aagttttaaa
    11881 gagttttagg cggaaaaatc gccttttttc tcttttatat cagtcactta catgtgtgac
    11941 cggttcccaa tgtacggctt tgggttccca atgtacgggt tccggttccc aatgtacggc
    12001 tttgggttcc caatgtacgt gctatccaca ggaaacagac cttttcgacc tttttcccct
    12061 gctagggcaa tttgccctag catctgctcc gtacattagg aaccggcgga tgcttcgccc
    12121 tcgatcaggt tgcggtagcg catgactagg atcgggccag cctgccccgc ctcctccttc
    12181 aaatcgtact ccggcaggtc atttgacccg atcagcttgc gcacggtgaa acagaacttc
    12241 ttgaactctc cggcgctgcc actgcgttcg tagatcgtct tgaacaacca tctggcttct
    12301 gccttgcctg cggcgcggcg tgccaggcgg tagagaaaac ggccgatgcc gggatcgatc
    12361 aaaaagtaat cggggtgaac cgtcagcacg tccgggttct tgccttctgt gatctcgcgg
    12421 tacatccaat cagctagctc gatctcgatg tactccggcc gcccggtttc gctctttacg
    12481 atcttgtagc ggctaatcaa ggcttcaccc tcggataccg tcaccaggcg gccgttcttg
    12541 gccttcttcg tacgctgcat ggcaacgtgc gtggtgttta accgaatgca ggtttctacc
    12601 aggtcgtctt tctgctttcc gccatcggct cgccggcaga acttgagtac gtccgcaacg
    12661 tgtggacgga acacgcggcc gggcttgtct cccttccctt cccggtatcg gttcatggat
    12721 tcggttagat gggaaaccgc catcagtacc aggtcgtaat cccacacact ggccatgccg
    12781 gccggccctg cggaaacctc tacgtgcccg tctggaagct cgtagcggat cacctcgcca
    12841 gctcgtcggt cacgcttcga cagacggaaa acggccacgt ccatgatgct gcgactatcg
    12901 cgggtgccca cgtcatagag catcggaacg aaaaaatctg gttgctcgtc gcccttgggc
    12961 ggcttcctaa tcgacggcgc accggctgcc ggcggttgcc gggattcttt gcggattcga
    13021 tcagcggccg cttgccacga ttcaccgggg cgtgcttctg cctcgatgcg ttgccgctgg
    13081 gcggcctgcg cggccttcaa cttctccacc aggtcatcac ccagcgccgc gccgatttgt
    13141 accgggccgg atggtttgcg accgctcacg ccgattcctc gggcttgggg gttccagtgc
    13201 cattgcaggg ccggcaggca acccagccgc ttacgcctgg ccaaccgccc gttcctccac
    13261 acatggggca ttccacggcg tcggtgcctg gttgttcttg attttccatg ccgcctcctt
    13321 tagccgctaa aattcatcta ctcatttatt catttgctca tttactctgg tagctgcgcg
    13381 atgtattcag atagcagctc ggtaatggtc ttgccttggc gtaccgcgta catcttcagc
    13441 ttggtgtgat cctccgccgg caactgaaag ttgacccgct tcatggctgg cgtgtctgcc
    13501 aggctggcca acgttgcagc cttgctgctg cgtgcgctcg gacggccggc acttagcgtg
    13561 tttgtgcttt tgctcatttt ctctttacct cattaactca aatgagtttt gatttaattt
    13621 cagcggccag cgcctggacc tcgcgggcag cgtcgccctc gggttctgat tcaagaacgg
    13681 ttgtgccggc ggcggcagtg cctgggtagc tcacgcgctg cgtgatacgg gactcaagaa
    13741 tgggcagctc gtacccggcc agcgcctcgg caacctcacc gccgatgcgc gtgcctttga
    13801 tcgcccgcga cacgacaaag gccgcttgta gccttccatc cgtgacctca atgcgctgct
    13861 taaccagctc caccaggtcg gcggtggccc atatgtcgta agggcttggc tgcaccggaa
    13921 tcagcacgaa gtcggctgcc ttgatcgcgg acacagccaa gtccgccgcc tggggcgctc
    13981 cgtcgatcac tacgaagtcg cgccggccga tggccttcac gtcgcggtca atcgtcgggc
    14041 ggtcgatgcc gacaacggtt agcggttgat cttcccgcac ggccgcccaa tcgcgggcac
    14101 tgccctgggg atcggaatcg actaacagaa catcggcccc ggcgagttgc agggcgcggg
    14161 ctagatgggt tgcgatggtc gtcttgcctg acccgccttt ctggttaagt acagcgataa
    14221 ccttcatgcg ttccccttgc gtatttgttt atttactcat cgcatcatat acgcagcgac
    14281 cgcatgacgc aagctgtttt actcaaatac acatcacctt tttagacggc ggcgctcggt
    14341 ttcttcagcg gccaagctgg ccggccaggc cgccagcttg gcatcagaca aaccggccag
    14401 gatttcatgc agccgcacgg ttgagacgtg cgcgggcggc tcgaacacgt acccggccgc
    14461 gatcatctcc gcctcgatct cttcggtaat gaaaaacggt tcgtcctggc cgtcctggtg
    14521 cggtttcatg cttgttcctc ttggcgttca ttctcggcgg ccgccagggc gtcggcctcg
    14581 gtcaatgcgt cctcacggaa ggcaccgcgc cgcctggcct cggtgggcgt cacttcctcg
    14641 ctgcgctcaa gtgcgcggta cagggtcgag cgatgcacgc caagcagtgc agccgcctct
    14701 ttcacggtgc ggccttcctg gtcgatcagc tcgcgggcgt gcgcgatctg tgccggggtg
    14761 agggtagggc gggggccaaa cttcacgcct cgggccttgg cggcctcgcg cccgctccgg
    14821 gtgcggtcga tgattaggga acgctcgaac tcggcaatgc cggcgaacac ggtcaacacc
    14881 atgcggccgg ccggcgtggt ggtgtcggcc cacggctctg ccaggctacg caggcccgcg
    14941 ccggcctcct ggatgcgctc ggcaatgtcc agtaggtcgc gggtgctgcg ggccaggcgg
    15001 tctagcctgg tcactgtcac aacgtcgcca gggcgtaggt ggtcaagcat cctggccagc
    15061 tccgggcggt cgcgcctggt gccggtgatc ttctcggaaa acagcttggt gcagccggcc
    15121 gcgtgcagtt cggcccgttg gttggtcaag tcctggtcgt cggtgctgac gcgggcatag
    15181 cccagcaggc cagcggcggc gctcttgttc atggcgtaat gtctccggtt ctagtcgcaa
    15241 gtattctact ttatgcgact aaaacacgcg acaagaaaac gccaggaaaa gggcagggcg
    15301 gcagcctgtc gcgtaactta ggacttgtgc gacatgtcgt tttcagaaga cggctgcact
    15361 gaacgtcaga agccgactgc actatagcag cggaggggtt ggatcaaagt actttgatcc
    15421 cgaggggaac cctgtggttg gcatgcacat acaaatggac gaacggataa accttttcac
    15481 gcccttttaa atatccgatt attctaataa acgctctttt ctcttaggtt tacccgccaa
    15541 tatatcctgt caaacactga tagtttaaac tgaaggcggg aaacgacaat ctgatccaag
    15601 ctcaagctgc tctagcattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
    15661 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
    15721 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagctt
    15781 cgacttgcct tccgcacaat acatcatttc ttcttagctt tttttcttct tcttcgttca
    15841 tacagttttt ttttgtttat cagcttacat tttcttgaac cgtagctttc gttttcttct
    15901 ttttaacttt ccattcggag tttttgtatc ttgtttcata gtttgtccca ggattagaat
    15961 gattaggcat cgaaccttca agaatttgat tgaataaaac atcttcattc ttaagatatg
    16021 aagataatct tcaaaaggcc cctgggaatc tgaaagaaga gaagcaggcc catttatatg
    16081 ggaaagaaca atagtatttc ttatataggc ccatttaagt tgaaaacaat cttcaaaagt
    16141 cccacatcgc ttagataaga aaacgaagct gagtttatat acagctagag tcgaagtagt
    16201 gattgggaga ccaacccagt ggacataagc ctgttcggtt cgtaagctgt aatgcaagta
    16261 gcgtatgcgc tcacgcaact ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc
    16321 agtggcggtt ttcatggctt gttatgactg tttttttggg gtacagtcta tgcctcgggc
    16381 atccaagcag caagcgcgtt acgccgtggg tcgatgtttg atgttatgga gcagcaacga
    16441 tgttacgcag cagggcagtc gccctaaaac aaagttaaac atcatggggg aagcggtgat
    16501 cgccgaagta tcgactcaac tatcagaggt agttggcgtc atcgagcgcc atctcgaacc
    16561 gacgttgctg gccgtacatt tgtacggctc cgcagtggat ggcggcctga agccacacag
    16621 tgatattgat ttgctggtta cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt
    16681 gatcaacgac cttttggaaa cttcggcttc ccctggagag agcgagattc tccgcgctgt
    16741 agaagtcacc attgttgtgc acgacgacat cattccgtgg cgttatccag ctaagcgcga
    16801 actgcaattt ggagaatggc agcgcaatga cattcttgca ggtatcttcg agccagccac
    16861 gatcgacatt gatctggcta tcttgctgac aaaagcaaga gaacatagcg ttgccttggt
    16921 aggtccagcg gcggaggaac tctttgatcc ggttcctgaa caggatctat ttgaggcgct
    16981 aaatgaaacc ttaacgctat ggaactcgcc gcccgactgg gctggcgatg agcgaaatgt
    17041 agtgcttacg ttgtcccgca tttggtacag cgcagtaacc ggcaaaatcg cgccgaagga
    17101 tgtcgctgcc gactgggcaa tggagcgcct gccggcccag tatcagcccg tcatacttga
    17161 agctagacag gcttatcttg gacaagaaga agatcgcttg gcctcgcgcg cagatcagtt
    17221 ggaagaattt gtccactacg tgaaaggcga gatcaccaag gtagtcggca aataatgtct
    17281 agctagaaat tcgttcaagc cgacgccgct tcgcggcgcg gcttaactca agcgttagat
    17341 gcactaagca cataattgct cacagccaaa ctatcaggtc aagtctgctt ttattatttt
    17401 taagcgtgca taataagccg gtctcg
//
