LOCUS       sequence-313815-       17420 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 CFP fluoresce..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17420)
  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 17420)
  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: pCUU
FEATURES             Location/Qualifiers
     source          1..17420
                     /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..8551)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIE"
     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    8990..9164
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    9242..9266
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             9691..10485
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      10572..11160
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11061..11080
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11314..11331
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    11346..11486
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(11412..11431)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     11572..11594
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      11830..12024
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(12090..13163)
                     /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(13592..14221)
                     /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    15522..15546
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     15734..15756
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     15748..15765
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     15749..15765
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             16478..17269
                     /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 caccctgacc tggggcgtgc agtgcttcag 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 atcagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa
     8521 ggccaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     8581 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     8641 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     8701 gttcgtgacc gccgccggga tcactctcgg catgaatcaa ttcaagcgtg aagagcaagc
     8761 aaggaaagct aaggtgaaca atgagaaaaa gacggaaata gtgaaaccag agagttgtag
     8821 caatgaagga gatgtcaagg atctgaaaag aaaggactct gaggatggaa acgagggtga
     8881 ggaagaagaa gcttcttcga aaccgaaaaa gccaaaagtt gctctttctc atcttcagga
     8941 cattgacgac acagaagctg atcaagaaga agagtaatct agtctcgagt ttctccataa
     9001 taatgtgtga gtagttccca gataagggaa ttagggttcc tatagggttt cgctcatgtg
     9061 ttgagcatat aagaaaccct tagtatgtat ttgtatttgt aaaatacttc tatcaataaa
     9121 atttctaatt cctaaaacca aaatccagta ctaaaatcca gatcccccga attaattcgg
     9181 cgttaattca gtacattaaa aacgtccgca atgtgttatt aagttgtcta agcgtcaatt
     9241 tgtttacacc acaatatatc ctgccaccag ccagccaaca gctccccgac cggcagctcg
     9301 gcacaaaatc accactcgat acaggcagcc catcagtccg ggacggcgtc agcgggagag
     9361 ccgttgtaag gcggcagact ttgctcatgt taccgatgct attcggaaga acggcaacta
     9421 agctgccggg tttgaaacac ggatgatctc gcggagggta gcatgttgat tgtaacgatg
     9481 acagagcgtt gctgcctgtg atcaccgcgg tttcaaaatc ggctccgtcg atactatgtt
     9541 atacgccaac tttgaaaaca actttgaaaa agctgttttc tggtatttaa ggttttagaa
     9601 tgcaaggaac agtgaattgg agttcgtctt gttataatta gcttcttggg gtatctttaa
     9661 atactgtaga aaagaggaag gaaataataa atggctaaaa tgagaatatc accggaattg
     9721 aaaaaactga tcgaaaaata ccgctgcgta aaagatacgg aaggaatgtc tcctgctaag
     9781 gtatataagc tggtgggaga aaatgaaaac ctatatttaa aaatgacgga cagccggtat
     9841 aaagggacca cctatgatgt ggaacgggaa aaggacatga tgctatggct ggaaggaaag
     9901 ctgcctgttc caaaggtcct gcactttgaa cggcatgatg gctggagcaa tctgctcatg
     9961 agtgaggccg atggcgtcct ttgctcggaa gagtatgaag atgaacaaag ccctgaaaag
    10021 attatcgagc tgtatgcgga gtgcatcagg ctctttcact ccatcgacat atcggattgt
    10081 ccctatacga atagcttaga cagccgctta gccgaattgg attacttact gaataacgat
    10141 ctggccgatg tggattgcga aaactgggaa gaagacactc catttaaaga tccgcgcgag
    10201 ctgtatgatt ttttaaagac ggaaaagccc gaagaggaac ttgtcttttc ccacggcgac
    10261 ctgggagaca gcaacatctt tgtgaaagat ggcaaagtaa gtggctttat tgatcttggg
    10321 agaagcggca gggcggacaa gtggtatgac attgccttct gcgtccggtc gatcagggag
    10381 gatatcgggg aagaacagta tgtcgagcta ttttttgact tactggggat caagcctgat
    10441 tgggagaaaa taaaatatta tattttactg gatgaattgt tttagtacct agaatgcatg
    10501 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
    10561 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
    10621 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
    10681 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
    10741 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
    10801 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
    10861 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
    10921 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
    10981 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
    11041 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
    11101 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
    11161 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
    11221 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
    11281 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
    11341 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg
    11401 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
    11461 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
    11521 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
    11581 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagggt gccttgatgt
    11641 gggcgccggc ggtcgagtgg cgacggcgcg gcttgtccgc gccctggtag attgcctggc
    11701 cgtaggccag ccatttttga gcggccagcg gccgcgatag gccgacgcga agcggcgggg
    11761 cgtagggagc gcagcgaccg aagggtaggc gctttttgca gctcttcggc tgtgcgctgg
    11821 ccagacagtt atgcacaggc caggcgggtt ttaagagttt taataagttt taaagagttt
    11881 taggcggaaa aatcgccttt tttctctttt atatcagtca cttacatgtg tgaccggttc
    11941 ccaatgtacg gctttgggtt cccaatgtac gggttccggt tcccaatgta cggctttggg
    12001 ttcccaatgt acgtgctatc cacaggaaac agaccttttc gacctttttc ccctgctagg
    12061 gcaatttgcc ctagcatctg ctccgtacat taggaaccgg cggatgcttc gccctcgatc
    12121 aggttgcggt agcgcatgac taggatcggg ccagcctgcc ccgcctcctc cttcaaatcg
    12181 tactccggca ggtcatttga cccgatcagc ttgcgcacgg tgaaacagaa cttcttgaac
    12241 tctccggcgc tgccactgcg ttcgtagatc gtcttgaaca accatctggc ttctgccttg
    12301 cctgcggcgc ggcgtgccag gcggtagaga aaacggccga tgccgggatc gatcaaaaag
    12361 taatcggggt gaaccgtcag cacgtccggg ttcttgcctt ctgtgatctc gcggtacatc
    12421 caatcagcta gctcgatctc gatgtactcc ggccgcccgg tttcgctctt tacgatcttg
    12481 tagcggctaa tcaaggcttc accctcggat accgtcacca ggcggccgtt cttggccttc
    12541 ttcgtacgct gcatggcaac gtgcgtggtg tttaaccgaa tgcaggtttc taccaggtcg
    12601 tctttctgct ttccgccatc ggctcgccgg cagaacttga gtacgtccgc aacgtgtgga
    12661 cggaacacgc ggccgggctt gtctcccttc ccttcccggt atcggttcat ggattcggtt
    12721 agatgggaaa ccgccatcag taccaggtcg taatcccaca cactggccat gccggccggc
    12781 cctgcggaaa cctctacgtg cccgtctgga agctcgtagc ggatcacctc gccagctcgt
    12841 cggtcacgct tcgacagacg gaaaacggcc acgtccatga tgctgcgact atcgcgggtg
    12901 cccacgtcat agagcatcgg aacgaaaaaa tctggttgct cgtcgccctt gggcggcttc
    12961 ctaatcgacg gcgcaccggc tgccggcggt tgccgggatt ctttgcggat tcgatcagcg
    13021 gccgcttgcc acgattcacc ggggcgtgct tctgcctcga tgcgttgccg ctgggcggcc
    13081 tgcgcggcct tcaacttctc caccaggtca tcacccagcg ccgcgccgat ttgtaccggg
    13141 ccggatggtt tgcgaccgct cacgccgatt cctcgggctt gggggttcca gtgccattgc
    13201 agggccggca ggcaacccag ccgcttacgc ctggccaacc gcccgttcct ccacacatgg
    13261 ggcattccac ggcgtcggtg cctggttgtt cttgattttc catgccgcct cctttagccg
    13321 ctaaaattca tctactcatt tattcatttg ctcatttact ctggtagctg cgcgatgtat
    13381 tcagatagca gctcggtaat ggtcttgcct tggcgtaccg cgtacatctt cagcttggtg
    13441 tgatcctccg ccggcaactg aaagttgacc cgcttcatgg ctggcgtgtc tgccaggctg
    13501 gccaacgttg cagccttgct gctgcgtgcg ctcggacggc cggcacttag cgtgtttgtg
    13561 cttttgctca ttttctcttt acctcattaa ctcaaatgag ttttgattta atttcagcgg
    13621 ccagcgcctg gacctcgcgg gcagcgtcgc cctcgggttc tgattcaaga acggttgtgc
    13681 cggcggcggc agtgcctggg tagctcacgc gctgcgtgat acgggactca agaatgggca
    13741 gctcgtaccc ggccagcgcc tcggcaacct caccgccgat gcgcgtgcct ttgatcgccc
    13801 gcgacacgac aaaggccgct tgtagccttc catccgtgac ctcaatgcgc tgcttaacca
    13861 gctccaccag gtcggcggtg gcccatatgt cgtaagggct tggctgcacc ggaatcagca
    13921 cgaagtcggc tgccttgatc gcggacacag ccaagtccgc cgcctggggc gctccgtcga
    13981 tcactacgaa gtcgcgccgg ccgatggcct tcacgtcgcg gtcaatcgtc gggcggtcga
    14041 tgccgacaac ggttagcggt tgatcttccc gcacggccgc ccaatcgcgg gcactgccct
    14101 ggggatcgga atcgactaac agaacatcgg ccccggcgag ttgcagggcg cgggctagat
    14161 gggttgcgat ggtcgtcttg cctgacccgc ctttctggtt aagtacagcg ataaccttca
    14221 tgcgttcccc ttgcgtattt gtttatttac tcatcgcatc atatacgcag cgaccgcatg
    14281 acgcaagctg ttttactcaa atacacatca cctttttaga cggcggcgct cggtttcttc
    14341 agcggccaag ctggccggcc aggccgccag cttggcatca gacaaaccgg ccaggatttc
    14401 atgcagccgc acggttgaga cgtgcgcggg cggctcgaac acgtacccgg ccgcgatcat
    14461 ctccgcctcg atctcttcgg taatgaaaaa cggttcgtcc tggccgtcct ggtgcggttt
    14521 catgcttgtt cctcttggcg ttcattctcg gcggccgcca gggcgtcggc ctcggtcaat
    14581 gcgtcctcac ggaaggcacc gcgccgcctg gcctcggtgg gcgtcacttc ctcgctgcgc
    14641 tcaagtgcgc ggtacagggt cgagcgatgc acgccaagca gtgcagccgc ctctttcacg
    14701 gtgcggcctt cctggtcgat cagctcgcgg gcgtgcgcga tctgtgccgg ggtgagggta
    14761 gggcgggggc caaacttcac gcctcgggcc ttggcggcct cgcgcccgct ccgggtgcgg
    14821 tcgatgatta gggaacgctc gaactcggca atgccggcga acacggtcaa caccatgcgg
    14881 ccggccggcg tggtggtgtc ggcccacggc tctgccaggc tacgcaggcc cgcgccggcc
    14941 tcctggatgc gctcggcaat gtccagtagg tcgcgggtgc tgcgggccag gcggtctagc
    15001 ctggtcactg tcacaacgtc gccagggcgt aggtggtcaa gcatcctggc cagctccggg
    15061 cggtcgcgcc tggtgccggt gatcttctcg gaaaacagct tggtgcagcc ggccgcgtgc
    15121 agttcggccc gttggttggt caagtcctgg tcgtcggtgc tgacgcgggc atagcccagc
    15181 aggccagcgg cggcgctctt gttcatggcg taatgtctcc ggttctagtc gcaagtattc
    15241 tactttatgc gactaaaaca cgcgacaaga aaacgccagg aaaagggcag ggcggcagcc
    15301 tgtcgcgtaa cttaggactt gtgcgacatg tcgttttcag aagacggctg cactgaacgt
    15361 cagaagccga ctgcactata gcagcggagg ggttggatca aagtactttg atcccgaggg
    15421 gaaccctgtg gttggcatgc acatacaaat ggacgaacgg ataaaccttt tcacgccctt
    15481 ttaaatatcc gattattcta ataaacgctc ttttctctta ggtttacccg ccaatatatc
    15541 ctgtcaaaca ctgatagttt aaactgaagg cgggaaacga caatctgatc caagctcaag
    15601 ctgctctagc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
    15661 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
    15721 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gcttcgactt
    15781 gccttccgca caatacatca tttcttctta gctttttttc ttcttcttcg ttcatacagt
    15841 ttttttttgt ttatcagctt acattttctt gaaccgtagc tttcgttttc ttctttttaa
    15901 ctttccattc ggagtttttg tatcttgttt catagtttgt cccaggatta gaatgattag
    15961 gcatcgaacc ttcaagaatt tgattgaata aaacatcttc attcttaaga tatgaagata
    16021 atcttcaaaa ggcccctggg aatctgaaag aagagaagca ggcccattta tatgggaaag
    16081 aacaatagta tttcttatat aggcccattt aagttgaaaa caatcttcaa aagtcccaca
    16141 tcgcttagat aagaaaacga agctgagttt atatacagct agagtcgaag tagtgattgg
    16201 gagaccaacc cagtggacat aagcctgttc ggttcgtaag ctgtaatgca agtagcgtat
    16261 gcgctcacgc aactggtcca gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc
    16321 ggttttcatg gcttgttatg actgtttttt tggggtacag tctatgcctc gggcatccaa
    16381 gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta tggagcagca acgatgttac
    16441 gcagcagggc agtcgcccta aaacaaagtt aaacatcatg ggggaagcgg tgatcgccga
    16501 agtatcgact caactatcag aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt
    16561 gctggccgta catttgtacg gctccgcagt ggatggcggc ctgaagccac acagtgatat
    16621 tgatttgctg gttacggtga ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa
    16681 cgaccttttg gaaacttcgg cttcccctgg agagagcgag attctccgcg ctgtagaagt
    16741 caccattgtt gtgcacgacg acatcattcc gtggcgttat ccagctaagc gcgaactgca
    16801 atttggagaa tggcagcgca atgacattct tgcaggtatc ttcgagccag ccacgatcga
    16861 cattgatctg gctatcttgc tgacaaaagc aagagaacat agcgttgcct tggtaggtcc
    16921 agcggcggag gaactctttg atccggttcc tgaacaggat ctatttgagg cgctaaatga
    16981 aaccttaacg ctatggaact cgccgcccga ctgggctggc gatgagcgaa atgtagtgct
    17041 tacgttgtcc cgcatttggt acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc
    17101 tgccgactgg gcaatggagc gcctgccggc ccagtatcag cccgtcatac ttgaagctag
    17161 acaggcttat cttggacaag aagaagatcg cttggcctcg cgcgcagatc agttggaaga
    17221 atttgtccac tacgtgaaag gcgagatcac caaggtagtc ggcaaataat gtctagctag
    17281 aaattcgttc aagccgacgc cgcttcgcgg cgcggcttaa ctcaagcgtt agatgcacta
    17341 agcacataat tgctcacagc caaactatca ggtcaagtct gcttttatta tttttaagcg
    17401 tgcataataa gccggtctcg
//
