LOCUS       sequence-313819-       16804 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Egg cell-specific promoter-controlled expression of 
            3×FLAG-NLS-zCas9-NLS. Contains gRNA scaffold for insertion of target
            sequence (U6-26 promoter). Selection of transgenic seeds by YFP 
            fluoresce..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16804)
  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 16804)
  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: pYEE
FEATURES             Location/Qualifiers
     source          1..16804
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        1..76
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             1646..1711
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1718..1738
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1763..5863
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             1763..5863
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     RBS             5776..5784
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             5864..5911
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(7402..7404,7405..7407,7408..8118)
                     /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(7447..7468)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7708..7727)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8055..8076
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    8374..8548
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    8626..8650
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             9075..9869
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      9956..10544
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10445..10464
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10698..10715
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    10730..10870
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(10796..10815)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     10956..10978
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      11214..11408
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(11474..12547)
                     /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(12976..13605)
                     /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    14906..14930
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     15118..15140
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     15132..15149
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     15133..15149
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             15862..16653
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
ORIGIN
        1 gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt
       61 ggcaccgagt cggtgctttt ttttgcaaaa ttttccagat cgatttcttc ttcctctgtt
      121 cttcggcgtt caatttctgg ggttttctct tcgttttctg taactgaaac ctaaaatttg
      181 acctaaaaaa aatctcaaat aatatgattc agtggttttg tacttttcag ttagttgagt
      241 tttgcagttc cgatgagata aaccaatacc atggttatac tagtgaataa aagcatttgc
      301 gtttggttta tcattgcgtt tatacaagga cagagatcca ctgagctgga atagcttaaa
      361 accattatca gaacaaaata aaccattttt tgttaagaat cagagcatag taaacaacag
      421 aaacaaccta agagaggtaa cttgtccaag aagatagcta attatatcta ttttataaaa
      481 gttatcatag tttgtaagtc acaaaagatg caaataacag agaaactagg agacttgaga
      541 atatacattc ttgtatattt gtattcgaga ttgtgaaaat ttgaccataa gtttaaattc
      601 ttaaaaagat atatctgatc taggtgatgg ttatagactg taattttacc acatgtttaa
      661 tgatggatag tgacacacat gacacatcga caacactata gcatcttatt tagattacaa
      721 catgaaattt ttctgtaata catgtctttg tacataattt aaaagtaatt cctaagaaat
      781 atatttatac aaggagttta aagaaaacat agcataaagt tcaatgagta gtaaaaacca
      841 tatacagtat atagcataaa gttcaatgag tttattacaa aagcattggt tcactttctg
      901 taacacgacg ttaaaccttc gtctccaata ggagcgctac tgattcaaca tgccaatata
      961 tactaaatac gtttctacag tcaaatgctt taacgtttca tgattaagtg actatttacc
     1021 gtcaatcctt tcccattcct cccactaatc caacttttta attactctta aatcaccact
     1081 aagctagtaa cgcctatcat gaattagctc tactaaatct agcaaccttt caaatttgca
     1141 gtattgcagg tgtctctgtg tctttaaaat agttgcctta tgatttcttc ggtttcaaga
     1201 tgatcaaata gttatagatt tcatgctcac acatgctcat tagatgtgta catactttac
     1261 ttacccaaat ctattttctc gcaaagattt tgatggtaaa gctgatttgg ttctattgaa
     1321 ctaaatcaaa cgagtttcag actgagtgat tctaatccgg cccattagcc cctaaacaga
     1381 cccactaatt acgcagcttt taatagagta attacaccta gtttacccac taaaccacta
     1441 agcactaatt atctcacaat ctaatgagct tccctcgtaa ttacttgggc tttcactcta
     1501 ccatttattt gtaacagtca agtctctact gtctctatat aaactctcta aagttaacac
     1561 acaattctca tcacaaacaa atcaaccaaa gcaacttcta ctctttcttc tttcgacctt
     1621 atcaatctgt tgagaaatct agatggatta caaggaccac gacggggatt acaaggacca
     1681 cgacattgat tacaaggatg atgatgacaa gatggctccg aagaagaaga ggaaggttgg
     1741 catccacggg gtgccagctg ctgacaagaa gtactcgatc ggcctcgata ttgggactaa
     1801 ctctgttggc tgggccgtga tcaccgacga gtacaaggtg ccctcaaaga agttcaaggt
     1861 cctgggcaac accgatcggc attccatcaa gaagaatctc attggcgctc tcctgttcga
     1921 cagcggcgag acggctgagg ctacgcggct caagcgcacc gcccgcaggc ggtacacgcg
     1981 caggaagaat cgcatctgct acctgcagga gattttctcc aacgagatgg cgaaggttga
     2041 cgattctttc ttccacaggc tggaggagtc attcctcgtg gaggaggata agaagcacga
     2101 gcggcatcca atcttcggca acattgtcga cgaggttgcc taccacgaga agtaccctac
     2161 gatctaccat ctgcggaaga agctcgtgga ctccacagat aaggcggacc tccgcctgat
     2221 ctacctcgct ctggcccaca tgattaagtt caggggccat ttcctgatcg agggggatct
     2281 caacccggac aatagcgatg ttgacaagct gttcatccag ctcgtgcaga cgtacaacca
     2341 gctcttcgag gagaacccca ttaatgcgtc aggcgtcgac gcgaaggcta tcctgtccgc
     2401 taggctctcg aagtctcggc gcctcgagaa cctgatcgcc cagctgccgg gcgagaagaa
     2461 gaacggcctg ttcgggaatc tcattgcgct cagcctgggg ctcacgccca acttcaagtc
     2521 gaatttcgat ctcgctgagg acgccaagct gcagctctcc aaggacacat acgacgatga
     2581 cctggataac ctcctggccc agatcggcga tcagtacgcg gacctgttcc tcgctgccaa
     2641 gaatctgtcg gacgccatcc tcctgtctga tattctcagg gtgaacaccg agattacgaa
     2701 ggctccgctc tcagcctcca tgatcaagcg ctacgacgag caccatcagg atctgaccct
     2761 cctgaaggcg ctggtcaggc agcagctccc cgagaagtac aaggagatct tcttcgatca
     2821 gtcgaagaac ggctacgctg ggtacattga cggcggggcc tctcaggagg agttctacaa
     2881 gttcatcaag ccgattctgg agaagatgga cggcacggag gagctgctgg tgaagctcaa
     2941 tcgcgaggac ctcctgagga agcagcggac attcgataac ggcagcatcc cacaccagat
     3001 tcatctcggg gagctgcacg ctatcctgag gaggcaggag gacttctacc ctttcctcaa
     3061 ggataaccgc gagaagatcg agaagattct gactttcagg atcccgtact acgtcggccc
     3121 actcgctagg ggcaactccc gcttcgcttg gatgacccgc aagtcagagg agacgatcac
     3181 gccgtggaac ttcgaggagg tggtcgacaa gggcgctagc gctcagtcgt tcatcgagag
     3241 gatgacgaat ttcgacaaga acctgccaaa tgagaaggtg ctccctaagc actcgctcct
     3301 gtacgagtac ttcacagtct acaacgagct gactaaggtg aagtatgtga ccgagggcat
     3361 gaggaagccg gctttcctgt ctggggagca gaagaaggcc atcgtggacc tcctgttcaa
     3421 gaccaaccgg aaggtcacgg ttaagcagct caaggaggac tacttcaaga agattgagtg
     3481 cttcgattcg gtcgagatct ctggcgttga ggaccgcttc aacgcctccc tggggaccta
     3541 ccacgatctc ctgaagatca ttaaggataa ggacttcctg gacaacgagg agaatgagga
     3601 tatcctcgag gacattgtgc tgacactcac tctgttcgag gaccgggaga tgatcgagga
     3661 gcgcctgaag acttacgccc atctcttcga tgacaaggtc atgaagcagc tcaagaggag
     3721 gaggtacacc ggctggggga ggctgagcag gaagctcatc aacggcattc gggacaagca
     3781 gtccgggaag acgatcctcg acttcctgaa gagcgatggc ttcgcgaacc gcaatttcat
     3841 gcagctgatt cacgatgaca gcctcacatt caaggaggat atccagaagg ctcaggtgag
     3901 cggccagggg gactcgctgc acgagcatat cgcgaacctc gctggctcgc cagctatcaa
     3961 gaaggggatt ctgcagaccg tgaaggttgt ggacgagctg gtgaaggtca tgggcaggca
     4021 caagcctgag aacatcgtca ttgagatggc ccgggagaat cagaccacgc agaagggcca
     4081 gaagaactca cgcgagagga tgaagaggat cgaggagggc attaaggagc tggggtccca
     4141 gatcctcaag gagcacccgg tggagaacac gcagctgcag aatgagaagc tctacctgta
     4201 ctacctccag aatggccgcg atatgtatgt ggaccaggag ctggatatta acaggctcag
     4261 cgattacgac gtcgatcata tcgttccaca gtcattcctg aaggatgact ccattgacaa
     4321 caaggtcctc accaggtcgg acaagaaccg gggcaagtct gataatgttc cttcagagga
     4381 ggtcgttaag aagatgaaga actactggcg ccagctcctg aatgccaagc tgatcacgca
     4441 gcggaagttc gataacctca caaaggctga gaggggcggg ctctctgagc tggacaaggc
     4501 gggcttcatc aagaggcagc tggtcgagac acggcagatc actaagcacg ttgcgcagat
     4561 tctcgactca cggatgaaca ctaagtacga tgagaatgac aagctgatcc gcgaggtgaa
     4621 ggtcatcacc ctgaagtcaa agctcgtctc cgacttcagg aaggatttcc agttctacaa
     4681 ggttcgggag atcaacaatt accaccatgc ccatgacgcg tacctgaacg cggtggtcgg
     4741 cacagctctg atcaagaagt acccaaagct cgagagcgag ttcgtgtacg gggactacaa
     4801 ggtttacgat gtgaggaaga tgatcgccaa gtcggagcag gagattggca aggctaccgc
     4861 caagtacttc ttctactcta acattatgaa tttcttcaag acagagatca ctctggccaa
     4921 tggcgagatc cggaagcgcc ccctcatcga gacgaacggc gagacggggg agatcgtgtg
     4981 ggacaagggc agggatttcg cgaccgtcag gaaggttctc tccatgccac aagtgaatat
     5041 cgtcaagaag acagaggtcc agactggcgg gttctctaag gagtcaattc tgcctaagcg
     5101 gaacagcgac aagctcatcg cccgcaagaa ggactgggat ccgaagaagt acggcgggtt
     5161 cgacagcccc actgtggcct actcggtcct ggttgtggcg aaggttgaga agggcaagtc
     5221 caagaagctc aagagcgtga aggagctgct ggggatcacg attatggagc gctccagctt
     5281 cgagaagaac ccgatcgatt tcctggaggc gaagggctac aaggaggtga agaaggacct
     5341 gatcattaag ctccccaagt actcactctt cgagctggag aacggcagga agcggatgct
     5401 ggcttccgct ggcgagctgc agaaggggaa cgagctggct ctgccgtcca agtatgtgaa
     5461 cttcctctac ctggcctccc actacgagaa gctcaagggc agccccgagg acaacgagca
     5521 gaagcagctg ttcgtcgagc agcacaagca ttacctcgac gagatcattg agcagatttc
     5581 cgagttctcc aagcgcgtga tcctggccga cgcgaatctg gataaggtcc tctccgcgta
     5641 caacaagcac cgcgacaagc caatcaggga gcaggctgag aatatcattc atctcttcac
     5701 cctgacgaac ctcggcgccc ctgctgcttt caagtacttc gacacaacta tcgatcgcaa
     5761 gaggtacaca agcactaagg aggtcctgga cgcgaccctc atccaccagt cgattaccgg
     5821 cctctacgag acgcgcatcg acctgtctca gctcgggggc gacaagcggc cagcggcgac
     5881 gaagaaggcg gggcaggcga agaagaagaa gtgagctcag agctttcgtt cgtatcatcg
     5941 gtttcgacaa cgttcgtcaa gttcaatgca tcagtttcat tgcgcacaca ccagaatcct
     6001 actgagtttg agtattatgg cattgggaaa actgtttttc ttgtaccatt tgttgtgctt
     6061 gtaatttact gtgtttttta ttcggttttc gctatcgaac tgtgaaatgg aaatggatgg
     6121 agaagagtta atgaatgata tggtcctttt gttcattctc aaattaatat tatttgtttt
     6181 ttctcttatt tgttgtgtgt tgaatttgaa attataagag atatgcaaac attttgtttt
     6241 gagtaaaaat gtgtcaaatc gtggcctcta atgaccgaag ttaatatgag gagtaaaaca
     6301 cttgtagttg taccattatg cttattcact aggcaacaaa tatattttca gacctagaaa
     6361 agctgcaaat gttactgaat acaagtatgt cctcttgtgt tttagacatt tatgaacttt
     6421 cctttatgta attttccaga atccttgtca gattctaatc attgctttat aattatagtt
     6481 atactcatgg atttgtagtt gagtatgaaa atatttttta atgcatttta tgacttgcca
     6541 attgattgac aacgaattcg ggcccattta aataggcgaa aggggtttag ggtttaagct
     6601 cagacttcta ttggagtaaa tgggacggtg tcacattttc cgttttggaa atgaactttg
     6661 ggctcacgtt atgggctatt agatatttga tgggctttct agtaaataca atataagtta
     6721 ttgggcttag tttaaataag cccatgttgg aaatatttga cacatgtctt ggctactagt
     6781 gctaaacatg caaccgaaca gttgtcgaga caagtcgcag catatacaat ggatcaaaca
     6841 cgcctagtgt cgccgcgtct cgctcatgtg tcaccttgtt tcctcgtttt tttttaattt
     6901 ttcataagtt cttttgtttt atcttcaata caaatttttg gctgtatctt gcaaactctt
     6961 cgatcatatc gccaatatac gtgaacactg gtgatctaat ttgttgtgtt aattgttaaa
     7021 tttagattct attctccggt ttaaaagtga attatatgta tcatggttaa aacattgtaa
     7081 gtaagatgat aataaaatga taaatttagt tgatggataa cgtgaagcaa aaaatgagat
     7141 agatacattt gattttgtcg tattttgaca tatgcggaga gtgagctacg cgcatgaaga
     7201 tcaagagaca cttgctcgag ctcacagagt gacgtgtaaa aagcttagac tgaagtcccc
     7261 atgcaaacct aatcctacgt ggctcaaacc acgagctcac ttgacaatat ataaactcct
     7321 cctaagtccc gttctcttca tccatctctc acaacaaaca aaaagaaaat ggttaagctc
     7381 agcaatctcc tcgttggatc tatggtgagc aagggcgagg agctgttcac cggggtggtg
     7441 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     7501 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     7561 ctgcccgtgc cctggcccac cctcgtgacc accttcggct acggcctgat gtgcttcgcc
     7621 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     7681 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     7741 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     7801 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     7861 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     7921 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     7981 gtgctgctgc ccgacaacca ctacctgagc taccagtccg ccctgagcaa agaccccaac
     8041 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     8101 atggacgagc tgtacaagaa acaattcaag cgtgaagagc aagcaaggaa agctaaggtg
     8161 aacaatgaga aaaagacgga aatagtgaaa ccagagagtt gtagcaatga aggagatgtc
     8221 aaggatctga aaagaaagga ctctgaggat ggaaacgagg gtgaggaaga agaagcttct
     8281 tcgaaaccga aaaagccaaa agttgctctt tctcatcttc aggacattga cgacacagaa
     8341 gctgatcaag aagaagagta atctagtctc gagtttctcc ataataatgt gtgagtagtt
     8401 cccagataag ggaattaggg ttcctatagg gtttcgctca tgtgttgagc atataagaaa
     8461 cccttagtat gtatttgtat ttgtaaaata cttctatcaa taaaatttct aattcctaaa
     8521 accaaaatcc agtactaaaa tccagatccc ccgaattaat tcggcgttaa ttcagtacat
     8581 taaaaacgtc cgcaatgtgt tattaagttg tctaagcgtc aatttgttta caccacaata
     8641 tatcctgcca ccagccagcc aacagctccc cgaccggcag ctcggcacaa aatcaccact
     8701 cgatacaggc agcccatcag tccgggacgg cgtcagcggg agagccgttg taaggcggca
     8761 gactttgctc atgttaccga tgctattcgg aagaacggca actaagctgc cgggtttgaa
     8821 acacggatga tctcgcggag ggtagcatgt tgattgtaac gatgacagag cgttgctgcc
     8881 tgtgatcacc gcggtttcaa aatcggctcc gtcgatacta tgttatacgc caactttgaa
     8941 aacaactttg aaaaagctgt tttctggtat ttaaggtttt agaatgcaag gaacagtgaa
     9001 ttggagttcg tcttgttata attagcttct tggggtatct ttaaatactg tagaaaagag
     9061 gaaggaaata ataaatggct aaaatgagaa tatcaccgga attgaaaaaa ctgatcgaaa
     9121 aataccgctg cgtaaaagat acggaaggaa tgtctcctgc taaggtatat aagctggtgg
     9181 gagaaaatga aaacctatat ttaaaaatga cggacagccg gtataaaggg accacctatg
     9241 atgtggaacg ggaaaaggac atgatgctat ggctggaagg aaagctgcct gttccaaagg
     9301 tcctgcactt tgaacggcat gatggctgga gcaatctgct catgagtgag gccgatggcg
     9361 tcctttgctc ggaagagtat gaagatgaac aaagccctga aaagattatc gagctgtatg
     9421 cggagtgcat caggctcttt cactccatcg acatatcgga ttgtccctat acgaatagct
     9481 tagacagccg cttagccgaa ttggattact tactgaataa cgatctggcc gatgtggatt
     9541 gcgaaaactg ggaagaagac actccattta aagatccgcg cgagctgtat gattttttaa
     9601 agacggaaaa gcccgaagag gaacttgtct tttcccacgg cgacctggga gacagcaaca
     9661 tctttgtgaa agatggcaaa gtaagtggct ttattgatct tgggagaagc ggcagggcgg
     9721 acaagtggta tgacattgcc ttctgcgtcc ggtcgatcag ggaggatatc ggggaagaac
     9781 agtatgtcga gctatttttt gacttactgg ggatcaagcc tgattgggag aaaataaaat
     9841 attatatttt actggatgaa ttgttttagt acctagaatg catgaccaaa atcccttaac
     9901 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     9961 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
    10021 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
    10081 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
    10141 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
    10201 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
    10261 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
    10321 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
    10381 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
    10441 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
    10501 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
    10561 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
    10621 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
    10681 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
    10741 attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa
    10801 tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt
    10861 catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
    10921 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
    10981 ttcaccgtca tcaccgaaac gcgcgaggca gggtgccttg atgtgggcgc cggcggtcga
    11041 gtggcgacgg cgcggcttgt ccgcgccctg gtagattgcc tggccgtagg ccagccattt
    11101 ttgagcggcc agcggccgcg ataggccgac gcgaagcggc ggggcgtagg gagcgcagcg
    11161 accgaagggt aggcgctttt tgcagctctt cggctgtgcg ctggccagac agttatgcac
    11221 aggccaggcg ggttttaaga gttttaataa gttttaaaga gttttaggcg gaaaaatcgc
    11281 cttttttctc ttttatatca gtcacttaca tgtgtgaccg gttcccaatg tacggctttg
    11341 ggttcccaat gtacgggttc cggttcccaa tgtacggctt tgggttccca atgtacgtgc
    11401 tatccacagg aaacagacct tttcgacctt tttcccctgc tagggcaatt tgccctagca
    11461 tctgctccgt acattaggaa ccggcggatg cttcgccctc gatcaggttg cggtagcgca
    11521 tgactaggat cgggccagcc tgccccgcct cctccttcaa atcgtactcc ggcaggtcat
    11581 ttgacccgat cagcttgcgc acggtgaaac agaacttctt gaactctccg gcgctgccac
    11641 tgcgttcgta gatcgtcttg aacaaccatc tggcttctgc cttgcctgcg gcgcggcgtg
    11701 ccaggcggta gagaaaacgg ccgatgccgg gatcgatcaa aaagtaatcg gggtgaaccg
    11761 tcagcacgtc cgggttcttg ccttctgtga tctcgcggta catccaatca gctagctcga
    11821 tctcgatgta ctccggccgc ccggtttcgc tctttacgat cttgtagcgg ctaatcaagg
    11881 cttcaccctc ggataccgtc accaggcggc cgttcttggc cttcttcgta cgctgcatgg
    11941 caacgtgcgt ggtgtttaac cgaatgcagg tttctaccag gtcgtctttc tgctttccgc
    12001 catcggctcg ccggcagaac ttgagtacgt ccgcaacgtg tggacggaac acgcggccgg
    12061 gcttgtctcc cttcccttcc cggtatcggt tcatggattc ggttagatgg gaaaccgcca
    12121 tcagtaccag gtcgtaatcc cacacactgg ccatgccggc cggccctgcg gaaacctcta
    12181 cgtgcccgtc tggaagctcg tagcggatca cctcgccagc tcgtcggtca cgcttcgaca
    12241 gacggaaaac ggccacgtcc atgatgctgc gactatcgcg ggtgcccacg tcatagagca
    12301 tcggaacgaa aaaatctggt tgctcgtcgc ccttgggcgg cttcctaatc gacggcgcac
    12361 cggctgccgg cggttgccgg gattctttgc ggattcgatc agcggccgct tgccacgatt
    12421 caccggggcg tgcttctgcc tcgatgcgtt gccgctgggc ggcctgcgcg gccttcaact
    12481 tctccaccag gtcatcaccc agcgccgcgc cgatttgtac cgggccggat ggtttgcgac
    12541 cgctcacgcc gattcctcgg gcttgggggt tccagtgcca ttgcagggcc ggcaggcaac
    12601 ccagccgctt acgcctggcc aaccgcccgt tcctccacac atggggcatt ccacggcgtc
    12661 ggtgcctggt tgttcttgat tttccatgcc gcctccttta gccgctaaaa ttcatctact
    12721 catttattca tttgctcatt tactctggta gctgcgcgat gtattcagat agcagctcgg
    12781 taatggtctt gccttggcgt accgcgtaca tcttcagctt ggtgtgatcc tccgccggca
    12841 actgaaagtt gacccgcttc atggctggcg tgtctgccag gctggccaac gttgcagcct
    12901 tgctgctgcg tgcgctcgga cggccggcac ttagcgtgtt tgtgcttttg ctcattttct
    12961 ctttacctca ttaactcaaa tgagttttga tttaatttca gcggccagcg cctggacctc
    13021 gcgggcagcg tcgccctcgg gttctgattc aagaacggtt gtgccggcgg cggcagtgcc
    13081 tgggtagctc acgcgctgcg tgatacggga ctcaagaatg ggcagctcgt acccggccag
    13141 cgcctcggca acctcaccgc cgatgcgcgt gcctttgatc gcccgcgaca cgacaaaggc
    13201 cgcttgtagc cttccatccg tgacctcaat gcgctgctta accagctcca ccaggtcggc
    13261 ggtggcccat atgtcgtaag ggcttggctg caccggaatc agcacgaagt cggctgcctt
    13321 gatcgcggac acagccaagt ccgccgcctg gggcgctccg tcgatcacta cgaagtcgcg
    13381 ccggccgatg gccttcacgt cgcggtcaat cgtcgggcgg tcgatgccga caacggttag
    13441 cggttgatct tcccgcacgg ccgcccaatc gcgggcactg ccctggggat cggaatcgac
    13501 taacagaaca tcggccccgg cgagttgcag ggcgcgggct agatgggttg cgatggtcgt
    13561 cttgcctgac ccgcctttct ggttaagtac agcgataacc ttcatgcgtt ccccttgcgt
    13621 atttgtttat ttactcatcg catcatatac gcagcgaccg catgacgcaa gctgttttac
    13681 tcaaatacac atcacctttt tagacggcgg cgctcggttt cttcagcggc caagctggcc
    13741 ggccaggccg ccagcttggc atcagacaaa ccggccagga tttcatgcag ccgcacggtt
    13801 gagacgtgcg cgggcggctc gaacacgtac ccggccgcga tcatctccgc ctcgatctct
    13861 tcggtaatga aaaacggttc gtcctggccg tcctggtgcg gtttcatgct tgttcctctt
    13921 ggcgttcatt ctcggcggcc gccagggcgt cggcctcggt caatgcgtcc tcacggaagg
    13981 caccgcgccg cctggcctcg gtgggcgtca cttcctcgct gcgctcaagt gcgcggtaca
    14041 gggtcgagcg atgcacgcca agcagtgcag ccgcctcttt cacggtgcgg ccttcctggt
    14101 cgatcagctc gcgggcgtgc gcgatctgtg ccggggtgag ggtagggcgg gggccaaact
    14161 tcacgcctcg ggccttggcg gcctcgcgcc cgctccgggt gcggtcgatg attagggaac
    14221 gctcgaactc ggcaatgccg gcgaacacgg tcaacaccat gcggccggcc ggcgtggtgg
    14281 tgtcggccca cggctctgcc aggctacgca ggcccgcgcc ggcctcctgg atgcgctcgg
    14341 caatgtccag taggtcgcgg gtgctgcggg ccaggcggtc tagcctggtc actgtcacaa
    14401 cgtcgccagg gcgtaggtgg tcaagcatcc tggccagctc cgggcggtcg cgcctggtgc
    14461 cggtgatctt ctcggaaaac agcttggtgc agccggccgc gtgcagttcg gcccgttggt
    14521 tggtcaagtc ctggtcgtcg gtgctgacgc gggcatagcc cagcaggcca gcggcggcgc
    14581 tcttgttcat ggcgtaatgt ctccggttct agtcgcaagt attctacttt atgcgactaa
    14641 aacacgcgac aagaaaacgc caggaaaagg gcagggcggc agcctgtcgc gtaacttagg
    14701 acttgtgcga catgtcgttt tcagaagacg gctgcactga acgtcagaag ccgactgcac
    14761 tatagcagcg gaggggttgg atcaaagtac tttgatcccg aggggaaccc tgtggttggc
    14821 atgcacatac aaatggacga acggataaac cttttcacgc ccttttaaat atccgattat
    14881 tctaataaac gctcttttct cttaggttta cccgccaata tatcctgtca aacactgata
    14941 gtttaaactg aaggcgggaa acgacaatct gatccaagct caagctgctc tagcattcgc
    15001 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
    15061 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
    15121 agtcacgacg ttgtaaaacg acggccagtg ccaagcttcg acttgccttc cgcacaatac
    15181 atcatttctt cttagctttt tttcttcttc ttcgttcata cagttttttt ttgtttatca
    15241 gcttacattt tcttgaaccg tagctttcgt tttcttcttt ttaactttcc attcggagtt
    15301 tttgtatctt gtttcatagt ttgtcccagg attagaatga ttaggcatcg aaccttcaag
    15361 aatttgattg aataaaacat cttcattctt aagatatgaa gataatcttc aaaaggcccc
    15421 tgggaatctg aaagaagaga agcaggccca tttatatggg aaagaacaat agtatttctt
    15481 atataggccc atttaagttg aaaacaatct tcaaaagtcc cacatcgctt agataagaaa
    15541 acgaagctga gtttatatac agctagagtc gaagtagtga ttgggagacc aacccagtgg
    15601 acataagcct gttcggttcg taagctgtaa tgcaagtagc gtatgcgctc acgcaactgg
    15661 tccagaacct tgaccgaacg cagcggtggt aacggcgcag tggcggtttt catggcttgt
    15721 tatgactgtt tttttggggt acagtctatg cctcgggcat ccaagcagca agcgcgttac
    15781 gccgtgggtc gatgtttgat gttatggagc agcaacgatg ttacgcagca gggcagtcgc
    15841 cctaaaacaa agttaaacat catgggggaa gcggtgatcg ccgaagtatc gactcaacta
    15901 tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc cgtacatttg
    15961 tacggctccg cagtggatgg cggcctgaag ccacacagtg atattgattt gctggttacg
    16021 gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga tcaacgacct tttggaaact
    16081 tcggcttccc ctggagagag cgagattctc cgcgctgtag aagtcaccat tgttgtgcac
    16141 gacgacatca ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg agaatggcag
    16201 cgcaatgaca ttcttgcagg tatcttcgag ccagccacga tcgacattga tctggctatc
    16261 ttgctgacaa aagcaagaga acatagcgtt gccttggtag gtccagcggc ggaggaactc
    16321 tttgatccgg ttcctgaaca ggatctattt gaggcgctaa atgaaacctt aacgctatgg
    16381 aactcgccgc ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt
    16441 tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg
    16501 gagcgcctgc cggcccagta tcagcccgtc atacttgaag ctagacaggc ttatcttgga
    16561 caagaagaag atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt ccactacgtg
    16621 aaaggcgaga tcaccaaggt agtcggcaaa taatgtctag ctagaaattc gttcaagccg
    16681 acgccgcttc gcggcgcggc ttaactcaag cgttagatgc actaagcaca taattgctca
    16741 cagccaaact atcaggtcaa gtctgctttt attattttta agcgtgcata ataagccggt
    16801 ctcg
//
