LOCUS       sequence-313821-       17435 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 YFP fluoresce..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17435)
  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 17435)
  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: pYUU
FEATURES             Location/Qualifiers
     source          1..17435
                     /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             join(8033..8035,8036..8038,8039..8749)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(8078..8099)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(8339..8358)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8686..8707
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    9005..9179
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    9257..9281
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             9706..10500
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      10587..11175
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11076..11095
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11329..11346
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    11361..11501
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(11427..11446)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     11587..11609
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      11845..12039
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(12105..13178)
                     /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(13607..14236)
                     /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    15537..15561
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     15749..15771
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     15763..15780
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     15764..15780
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             16493..17284
                     /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 ctatggtgag
     8041 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     8101 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     8161 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     8221 caccttcggc tacggcctga tgtgcttcgc ccgctacccc gaccacatga agcagcacga
     8281 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     8341 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     8401 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     8461 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     8521 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     8581 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     8641 ctaccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     8701 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaaga aacaattcaa
     8761 gcgtgaagag caagcaagga aagctaaggt gaacaatgag aaaaagacgg aaatagtgaa
     8821 accagagagt tgtagcaatg aaggagatgt caaggatctg aaaagaaagg actctgagga
     8881 tggaaacgag ggtgaggaag aagaagcttc ttcgaaaccg aaaaagccaa aagttgctct
     8941 ttctcatctt caggacattg acgacacaga agctgatcaa gaagaagagt aatctagtct
     9001 cgagtttctc cataataatg tgtgagtagt tcccagataa gggaattagg gttcctatag
     9061 ggtttcgctc atgtgttgag catataagaa acccttagta tgtatttgta tttgtaaaat
     9121 acttctatca ataaaatttc taattcctaa aaccaaaatc cagtactaaa atccagatcc
     9181 cccgaattaa ttcggcgtta attcagtaca ttaaaaacgt ccgcaatgtg ttattaagtt
     9241 gtctaagcgt caatttgttt acaccacaat atatcctgcc accagccagc caacagctcc
     9301 ccgaccggca gctcggcaca aaatcaccac tcgatacagg cagcccatca gtccgggacg
     9361 gcgtcagcgg gagagccgtt gtaaggcggc agactttgct catgttaccg atgctattcg
     9421 gaagaacggc aactaagctg ccgggtttga aacacggatg atctcgcgga gggtagcatg
     9481 ttgattgtaa cgatgacaga gcgttgctgc ctgtgatcac cgcggtttca aaatcggctc
     9541 cgtcgatact atgttatacg ccaactttga aaacaacttt gaaaaagctg ttttctggta
     9601 tttaaggttt tagaatgcaa ggaacagtga attggagttc gtcttgttat aattagcttc
     9661 ttggggtatc tttaaatact gtagaaaaga ggaaggaaat aataaatggc taaaatgaga
     9721 atatcaccgg aattgaaaaa actgatcgaa aaataccgct gcgtaaaaga tacggaagga
     9781 atgtctcctg ctaaggtata taagctggtg ggagaaaatg aaaacctata tttaaaaatg
     9841 acggacagcc ggtataaagg gaccacctat gatgtggaac gggaaaagga catgatgcta
     9901 tggctggaag gaaagctgcc tgttccaaag gtcctgcact ttgaacggca tgatggctgg
     9961 agcaatctgc tcatgagtga ggccgatggc gtcctttgct cggaagagta tgaagatgaa
    10021 caaagccctg aaaagattat cgagctgtat gcggagtgca tcaggctctt tcactccatc
    10081 gacatatcgg attgtcccta tacgaatagc ttagacagcc gcttagccga attggattac
    10141 ttactgaata acgatctggc cgatgtggat tgcgaaaact gggaagaaga cactccattt
    10201 aaagatccgc gcgagctgta tgatttttta aagacggaaa agcccgaaga ggaacttgtc
    10261 ttttcccacg gcgacctggg agacagcaac atctttgtga aagatggcaa agtaagtggc
    10321 tttattgatc ttgggagaag cggcagggcg gacaagtggt atgacattgc cttctgcgtc
    10381 cggtcgatca gggaggatat cggggaagaa cagtatgtcg agctattttt tgacttactg
    10441 gggatcaagc ctgattggga gaaaataaaa tattatattt tactggatga attgttttag
    10501 tacctagaat gcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
    10561 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
    10621 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
    10681 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
    10741 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
    10801 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
    10861 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
    10921 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
    10981 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
    11041 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
    11101 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
    11161 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
    11221 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
    11281 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
    11341 agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
    11401 tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
    11461 tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac
    11521 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
    11581 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc
    11641 agggtgcctt gatgtgggcg ccggcggtcg agtggcgacg gcgcggcttg tccgcgccct
    11701 ggtagattgc ctggccgtag gccagccatt tttgagcggc cagcggccgc gataggccga
    11761 cgcgaagcgg cggggcgtag ggagcgcagc gaccgaaggg taggcgcttt ttgcagctct
    11821 tcggctgtgc gctggccaga cagttatgca caggccaggc gggttttaag agttttaata
    11881 agttttaaag agttttaggc ggaaaaatcg ccttttttct cttttatatc agtcacttac
    11941 atgtgtgacc ggttcccaat gtacggcttt gggttcccaa tgtacgggtt ccggttccca
    12001 atgtacggct ttgggttccc aatgtacgtg ctatccacag gaaacagacc ttttcgacct
    12061 ttttcccctg ctagggcaat ttgccctagc atctgctccg tacattagga accggcggat
    12121 gcttcgccct cgatcaggtt gcggtagcgc atgactagga tcgggccagc ctgccccgcc
    12181 tcctccttca aatcgtactc cggcaggtca tttgacccga tcagcttgcg cacggtgaaa
    12241 cagaacttct tgaactctcc ggcgctgcca ctgcgttcgt agatcgtctt gaacaaccat
    12301 ctggcttctg ccttgcctgc ggcgcggcgt gccaggcggt agagaaaacg gccgatgccg
    12361 ggatcgatca aaaagtaatc ggggtgaacc gtcagcacgt ccgggttctt gccttctgtg
    12421 atctcgcggt acatccaatc agctagctcg atctcgatgt actccggccg cccggtttcg
    12481 ctctttacga tcttgtagcg gctaatcaag gcttcaccct cggataccgt caccaggcgg
    12541 ccgttcttgg ccttcttcgt acgctgcatg gcaacgtgcg tggtgtttaa ccgaatgcag
    12601 gtttctacca ggtcgtcttt ctgctttccg ccatcggctc gccggcagaa cttgagtacg
    12661 tccgcaacgt gtggacggaa cacgcggccg ggcttgtctc ccttcccttc ccggtatcgg
    12721 ttcatggatt cggttagatg ggaaaccgcc atcagtacca ggtcgtaatc ccacacactg
    12781 gccatgccgg ccggccctgc ggaaacctct acgtgcccgt ctggaagctc gtagcggatc
    12841 acctcgccag ctcgtcggtc acgcttcgac agacggaaaa cggccacgtc catgatgctg
    12901 cgactatcgc gggtgcccac gtcatagagc atcggaacga aaaaatctgg ttgctcgtcg
    12961 cccttgggcg gcttcctaat cgacggcgca ccggctgccg gcggttgccg ggattctttg
    13021 cggattcgat cagcggccgc ttgccacgat tcaccggggc gtgcttctgc ctcgatgcgt
    13081 tgccgctggg cggcctgcgc ggccttcaac ttctccacca ggtcatcacc cagcgccgcg
    13141 ccgatttgta ccgggccgga tggtttgcga ccgctcacgc cgattcctcg ggcttggggg
    13201 ttccagtgcc attgcagggc cggcaggcaa cccagccgct tacgcctggc caaccgcccg
    13261 ttcctccaca catggggcat tccacggcgt cggtgcctgg ttgttcttga ttttccatgc
    13321 cgcctccttt agccgctaaa attcatctac tcatttattc atttgctcat ttactctggt
    13381 agctgcgcga tgtattcaga tagcagctcg gtaatggtct tgccttggcg taccgcgtac
    13441 atcttcagct tggtgtgatc ctccgccggc aactgaaagt tgacccgctt catggctggc
    13501 gtgtctgcca ggctggccaa cgttgcagcc ttgctgctgc gtgcgctcgg acggccggca
    13561 cttagcgtgt ttgtgctttt gctcattttc tctttacctc attaactcaa atgagttttg
    13621 atttaatttc agcggccagc gcctggacct cgcgggcagc gtcgccctcg ggttctgatt
    13681 caagaacggt tgtgccggcg gcggcagtgc ctgggtagct cacgcgctgc gtgatacggg
    13741 actcaagaat gggcagctcg tacccggcca gcgcctcggc aacctcaccg ccgatgcgcg
    13801 tgcctttgat cgcccgcgac acgacaaagg ccgcttgtag ccttccatcc gtgacctcaa
    13861 tgcgctgctt aaccagctcc accaggtcgg cggtggccca tatgtcgtaa gggcttggct
    13921 gcaccggaat cagcacgaag tcggctgcct tgatcgcgga cacagccaag tccgccgcct
    13981 ggggcgctcc gtcgatcact acgaagtcgc gccggccgat ggccttcacg tcgcggtcaa
    14041 tcgtcgggcg gtcgatgccg acaacggtta gcggttgatc ttcccgcacg gccgcccaat
    14101 cgcgggcact gccctgggga tcggaatcga ctaacagaac atcggccccg gcgagttgca
    14161 gggcgcgggc tagatgggtt gcgatggtcg tcttgcctga cccgcctttc tggttaagta
    14221 cagcgataac cttcatgcgt tccccttgcg tatttgttta tttactcatc gcatcatata
    14281 cgcagcgacc gcatgacgca agctgtttta ctcaaataca catcaccttt ttagacggcg
    14341 gcgctcggtt tcttcagcgg ccaagctggc cggccaggcc gccagcttgg catcagacaa
    14401 accggccagg atttcatgca gccgcacggt tgagacgtgc gcgggcggct cgaacacgta
    14461 cccggccgcg atcatctccg cctcgatctc ttcggtaatg aaaaacggtt cgtcctggcc
    14521 gtcctggtgc ggtttcatgc ttgttcctct tggcgttcat tctcggcggc cgccagggcg
    14581 tcggcctcgg tcaatgcgtc ctcacggaag gcaccgcgcc gcctggcctc ggtgggcgtc
    14641 acttcctcgc tgcgctcaag tgcgcggtac agggtcgagc gatgcacgcc aagcagtgca
    14701 gccgcctctt tcacggtgcg gccttcctgg tcgatcagct cgcgggcgtg cgcgatctgt
    14761 gccggggtga gggtagggcg ggggccaaac ttcacgcctc gggccttggc ggcctcgcgc
    14821 ccgctccggg tgcggtcgat gattagggaa cgctcgaact cggcaatgcc ggcgaacacg
    14881 gtcaacacca tgcggccggc cggcgtggtg gtgtcggccc acggctctgc caggctacgc
    14941 aggcccgcgc cggcctcctg gatgcgctcg gcaatgtcca gtaggtcgcg ggtgctgcgg
    15001 gccaggcggt ctagcctggt cactgtcaca acgtcgccag ggcgtaggtg gtcaagcatc
    15061 ctggccagct ccgggcggtc gcgcctggtg ccggtgatct tctcggaaaa cagcttggtg
    15121 cagccggccg cgtgcagttc ggcccgttgg ttggtcaagt cctggtcgtc ggtgctgacg
    15181 cgggcatagc ccagcaggcc agcggcggcg ctcttgttca tggcgtaatg tctccggttc
    15241 tagtcgcaag tattctactt tatgcgacta aaacacgcga caagaaaacg ccaggaaaag
    15301 ggcagggcgg cagcctgtcg cgtaacttag gacttgtgcg acatgtcgtt ttcagaagac
    15361 ggctgcactg aacgtcagaa gccgactgca ctatagcagc ggaggggttg gatcaaagta
    15421 ctttgatccc gaggggaacc ctgtggttgg catgcacata caaatggacg aacggataaa
    15481 ccttttcacg cccttttaaa tatccgatta ttctaataaa cgctcttttc tcttaggttt
    15541 acccgccaat atatcctgtc aaacactgat agtttaaact gaaggcggga aacgacaatc
    15601 tgatccaagc tcaagctgct ctagcattcg ccattcaggc tgcgcaactg ttgggaaggg
    15661 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
    15721 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
    15781 gccaagcttc gacttgcctt ccgcacaata catcatttct tcttagcttt ttttcttctt
    15841 cttcgttcat acagtttttt tttgtttatc agcttacatt ttcttgaacc gtagctttcg
    15901 ttttcttctt tttaactttc cattcggagt ttttgtatct tgtttcatag tttgtcccag
    15961 gattagaatg attaggcatc gaaccttcaa gaatttgatt gaataaaaca tcttcattct
    16021 taagatatga agataatctt caaaaggccc ctgggaatct gaaagaagag aagcaggccc
    16081 atttatatgg gaaagaacaa tagtatttct tatataggcc catttaagtt gaaaacaatc
    16141 ttcaaaagtc ccacatcgct tagataagaa aacgaagctg agtttatata cagctagagt
    16201 cgaagtagtg attgggagac caacccagtg gacataagcc tgttcggttc gtaagctgta
    16261 atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg
    16321 taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgggg tacagtctat
    16381 gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag
    16441 cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca tcatggggga
    16501 agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca
    16561 tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa
    16621 gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg
    16681 gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct
    16741 ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc
    16801 taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga
    16861 gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt
    16921 tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt
    16981 tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga
    17041 gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc
    17101 gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt
    17161 catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc
    17221 agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg tagtcggcaa
    17281 ataatgtcta gctagaaatt cgttcaagcc gacgccgctt cgcggcgcgg cttaactcaa
    17341 gcgttagatg cactaagcac ataattgctc acagccaaac tatcaggtca agtctgcttt
    17401 tattattttt aagcgtgcat aataagccgg tctcg
//
