LOCUS       sequence-313813-       16789 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 CFP 
            fluoresce..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16789)
  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 16789)
  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: pCEE
FEATURES             Location/Qualifiers
     source          1..16789
                     /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..7920)
                     /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(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    8359..8533
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    8611..8635
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             9060..9854
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      9941..10529
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10430..10449
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10683..10700
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    10715..10855
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(10781..10800)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     10941..10963
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      11199..11393
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(11459..12532)
                     /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(12961..13590)
                     /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    14891..14915
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     15103..15125
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     15117..15134
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     15118..15134
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             15847..16638
                     /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 accctgacct ggggcgtgca gtgcttcagc
     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 tcagccacaa cgtctatatc
     7861 accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag
     7921 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     7981 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     8041 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     8101 atgaatcaat tcaagcgtga agagcaagca aggaaagcta aggtgaacaa tgagaaaaag
     8161 acggaaatag tgaaaccaga gagttgtagc aatgaaggag atgtcaagga tctgaaaaga
     8221 aaggactctg aggatggaaa cgagggtgag gaagaagaag cttcttcgaa accgaaaaag
     8281 ccaaaagttg ctctttctca tcttcaggac attgacgaca cagaagctga tcaagaagaa
     8341 gagtaatcta gtctcgagtt tctccataat aatgtgtgag tagttcccag ataagggaat
     8401 tagggttcct atagggtttc gctcatgtgt tgagcatata agaaaccctt agtatgtatt
     8461 tgtatttgta aaatacttct atcaataaaa tttctaattc ctaaaaccaa aatccagtac
     8521 taaaatccag atcccccgaa ttaattcggc gttaattcag tacattaaaa acgtccgcaa
     8581 tgtgttatta agttgtctaa gcgtcaattt gtttacacca caatatatcc tgccaccagc
     8641 cagccaacag ctccccgacc ggcagctcgg cacaaaatca ccactcgata caggcagccc
     8701 atcagtccgg gacggcgtca gcgggagagc cgttgtaagg cggcagactt tgctcatgtt
     8761 accgatgcta ttcggaagaa cggcaactaa gctgccgggt ttgaaacacg gatgatctcg
     8821 cggagggtag catgttgatt gtaacgatga cagagcgttg ctgcctgtga tcaccgcggt
     8881 ttcaaaatcg gctccgtcga tactatgtta tacgccaact ttgaaaacaa ctttgaaaaa
     8941 gctgttttct ggtatttaag gttttagaat gcaaggaaca gtgaattgga gttcgtcttg
     9001 ttataattag cttcttgggg tatctttaaa tactgtagaa aagaggaagg aaataataaa
     9061 tggctaaaat gagaatatca ccggaattga aaaaactgat cgaaaaatac cgctgcgtaa
     9121 aagatacgga aggaatgtct cctgctaagg tatataagct ggtgggagaa aatgaaaacc
     9181 tatatttaaa aatgacggac agccggtata aagggaccac ctatgatgtg gaacgggaaa
     9241 aggacatgat gctatggctg gaaggaaagc tgcctgttcc aaaggtcctg cactttgaac
     9301 ggcatgatgg ctggagcaat ctgctcatga gtgaggccga tggcgtcctt tgctcggaag
     9361 agtatgaaga tgaacaaagc cctgaaaaga ttatcgagct gtatgcggag tgcatcaggc
     9421 tctttcactc catcgacata tcggattgtc cctatacgaa tagcttagac agccgcttag
     9481 ccgaattgga ttacttactg aataacgatc tggccgatgt ggattgcgaa aactgggaag
     9541 aagacactcc atttaaagat ccgcgcgagc tgtatgattt tttaaagacg gaaaagcccg
     9601 aagaggaact tgtcttttcc cacggcgacc tgggagacag caacatcttt gtgaaagatg
     9661 gcaaagtaag tggctttatt gatcttggga gaagcggcag ggcggacaag tggtatgaca
     9721 ttgccttctg cgtccggtcg atcagggagg atatcgggga agaacagtat gtcgagctat
     9781 tttttgactt actggggatc aagcctgatt gggagaaaat aaaatattat attttactgg
     9841 atgaattgtt ttagtaccta gaatgcatga ccaaaatccc ttaacgtgag ttttcgttcc
     9901 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     9961 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
    10021 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
    10081 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
    10141 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
    10201 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
    10261 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
    10321 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
    10381 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
    10441 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
    10501 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
    10561 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
    10621 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
    10681 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
    10741 atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg
    10801 catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg
    10861 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
    10921 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
    10981 gaaacgcgcg aggcagggtg ccttgatgtg ggcgccggcg gtcgagtggc gacggcgcgg
    11041 cttgtccgcg ccctggtaga ttgcctggcc gtaggccagc catttttgag cggccagcgg
    11101 ccgcgatagg ccgacgcgaa gcggcggggc gtagggagcg cagcgaccga agggtaggcg
    11161 ctttttgcag ctcttcggct gtgcgctggc cagacagtta tgcacaggcc aggcgggttt
    11221 taagagtttt aataagtttt aaagagtttt aggcggaaaa atcgcctttt ttctctttta
    11281 tatcagtcac ttacatgtgt gaccggttcc caatgtacgg ctttgggttc ccaatgtacg
    11341 ggttccggtt cccaatgtac ggctttgggt tcccaatgta cgtgctatcc acaggaaaca
    11401 gaccttttcg acctttttcc cctgctaggg caatttgccc tagcatctgc tccgtacatt
    11461 aggaaccggc ggatgcttcg ccctcgatca ggttgcggta gcgcatgact aggatcgggc
    11521 cagcctgccc cgcctcctcc ttcaaatcgt actccggcag gtcatttgac ccgatcagct
    11581 tgcgcacggt gaaacagaac ttcttgaact ctccggcgct gccactgcgt tcgtagatcg
    11641 tcttgaacaa ccatctggct tctgccttgc ctgcggcgcg gcgtgccagg cggtagagaa
    11701 aacggccgat gccgggatcg atcaaaaagt aatcggggtg aaccgtcagc acgtccgggt
    11761 tcttgccttc tgtgatctcg cggtacatcc aatcagctag ctcgatctcg atgtactccg
    11821 gccgcccggt ttcgctcttt acgatcttgt agcggctaat caaggcttca ccctcggata
    11881 ccgtcaccag gcggccgttc ttggccttct tcgtacgctg catggcaacg tgcgtggtgt
    11941 ttaaccgaat gcaggtttct accaggtcgt ctttctgctt tccgccatcg gctcgccggc
    12001 agaacttgag tacgtccgca acgtgtggac ggaacacgcg gccgggcttg tctcccttcc
    12061 cttcccggta tcggttcatg gattcggtta gatgggaaac cgccatcagt accaggtcgt
    12121 aatcccacac actggccatg ccggccggcc ctgcggaaac ctctacgtgc ccgtctggaa
    12181 gctcgtagcg gatcacctcg ccagctcgtc ggtcacgctt cgacagacgg aaaacggcca
    12241 cgtccatgat gctgcgacta tcgcgggtgc ccacgtcata gagcatcgga acgaaaaaat
    12301 ctggttgctc gtcgcccttg ggcggcttcc taatcgacgg cgcaccggct gccggcggtt
    12361 gccgggattc tttgcggatt cgatcagcgg ccgcttgcca cgattcaccg gggcgtgctt
    12421 ctgcctcgat gcgttgccgc tgggcggcct gcgcggcctt caacttctcc accaggtcat
    12481 cacccagcgc cgcgccgatt tgtaccgggc cggatggttt gcgaccgctc acgccgattc
    12541 ctcgggcttg ggggttccag tgccattgca gggccggcag gcaacccagc cgcttacgcc
    12601 tggccaaccg cccgttcctc cacacatggg gcattccacg gcgtcggtgc ctggttgttc
    12661 ttgattttcc atgccgcctc ctttagccgc taaaattcat ctactcattt attcatttgc
    12721 tcatttactc tggtagctgc gcgatgtatt cagatagcag ctcggtaatg gtcttgcctt
    12781 ggcgtaccgc gtacatcttc agcttggtgt gatcctccgc cggcaactga aagttgaccc
    12841 gcttcatggc tggcgtgtct gccaggctgg ccaacgttgc agccttgctg ctgcgtgcgc
    12901 tcggacggcc ggcacttagc gtgtttgtgc ttttgctcat tttctcttta cctcattaac
    12961 tcaaatgagt tttgatttaa tttcagcggc cagcgcctgg acctcgcggg cagcgtcgcc
    13021 ctcgggttct gattcaagaa cggttgtgcc ggcggcggca gtgcctgggt agctcacgcg
    13081 ctgcgtgata cgggactcaa gaatgggcag ctcgtacccg gccagcgcct cggcaacctc
    13141 accgccgatg cgcgtgcctt tgatcgcccg cgacacgaca aaggccgctt gtagccttcc
    13201 atccgtgacc tcaatgcgct gcttaaccag ctccaccagg tcggcggtgg cccatatgtc
    13261 gtaagggctt ggctgcaccg gaatcagcac gaagtcggct gccttgatcg cggacacagc
    13321 caagtccgcc gcctggggcg ctccgtcgat cactacgaag tcgcgccggc cgatggcctt
    13381 cacgtcgcgg tcaatcgtcg ggcggtcgat gccgacaacg gttagcggtt gatcttcccg
    13441 cacggccgcc caatcgcggg cactgccctg gggatcggaa tcgactaaca gaacatcggc
    13501 cccggcgagt tgcagggcgc gggctagatg ggttgcgatg gtcgtcttgc ctgacccgcc
    13561 tttctggtta agtacagcga taaccttcat gcgttcccct tgcgtatttg tttatttact
    13621 catcgcatca tatacgcagc gaccgcatga cgcaagctgt tttactcaaa tacacatcac
    13681 ctttttagac ggcggcgctc ggtttcttca gcggccaagc tggccggcca ggccgccagc
    13741 ttggcatcag acaaaccggc caggatttca tgcagccgca cggttgagac gtgcgcgggc
    13801 ggctcgaaca cgtacccggc cgcgatcatc tccgcctcga tctcttcggt aatgaaaaac
    13861 ggttcgtcct ggccgtcctg gtgcggtttc atgcttgttc ctcttggcgt tcattctcgg
    13921 cggccgccag ggcgtcggcc tcggtcaatg cgtcctcacg gaaggcaccg cgccgcctgg
    13981 cctcggtggg cgtcacttcc tcgctgcgct caagtgcgcg gtacagggtc gagcgatgca
    14041 cgccaagcag tgcagccgcc tctttcacgg tgcggccttc ctggtcgatc agctcgcggg
    14101 cgtgcgcgat ctgtgccggg gtgagggtag ggcgggggcc aaacttcacg cctcgggcct
    14161 tggcggcctc gcgcccgctc cgggtgcggt cgatgattag ggaacgctcg aactcggcaa
    14221 tgccggcgaa cacggtcaac accatgcggc cggccggcgt ggtggtgtcg gcccacggct
    14281 ctgccaggct acgcaggccc gcgccggcct cctggatgcg ctcggcaatg tccagtaggt
    14341 cgcgggtgct gcgggccagg cggtctagcc tggtcactgt cacaacgtcg ccagggcgta
    14401 ggtggtcaag catcctggcc agctccgggc ggtcgcgcct ggtgccggtg atcttctcgg
    14461 aaaacagctt ggtgcagccg gccgcgtgca gttcggcccg ttggttggtc aagtcctggt
    14521 cgtcggtgct gacgcgggca tagcccagca ggccagcggc ggcgctcttg ttcatggcgt
    14581 aatgtctccg gttctagtcg caagtattct actttatgcg actaaaacac gcgacaagaa
    14641 aacgccagga aaagggcagg gcggcagcct gtcgcgtaac ttaggacttg tgcgacatgt
    14701 cgttttcaga agacggctgc actgaacgtc agaagccgac tgcactatag cagcggaggg
    14761 gttggatcaa agtactttga tcccgagggg aaccctgtgg ttggcatgca catacaaatg
    14821 gacgaacgga taaacctttt cacgcccttt taaatatccg attattctaa taaacgctct
    14881 tttctcttag gtttacccgc caatatatcc tgtcaaacac tgatagttta aactgaaggc
    14941 gggaaacgac aatctgatcc aagctcaagc tgctctagca ttcgccattc aggctgcgca
    15001 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
    15061 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
    15121 aaacgacggc cagtgccaag cttcgacttg ccttccgcac aatacatcat ttcttcttag
    15181 ctttttttct tcttcttcgt tcatacagtt tttttttgtt tatcagctta cattttcttg
    15241 aaccgtagct ttcgttttct tctttttaac tttccattcg gagtttttgt atcttgtttc
    15301 atagtttgtc ccaggattag aatgattagg catcgaacct tcaagaattt gattgaataa
    15361 aacatcttca ttcttaagat atgaagataa tcttcaaaag gcccctggga atctgaaaga
    15421 agagaagcag gcccatttat atgggaaaga acaatagtat ttcttatata ggcccattta
    15481 agttgaaaac aatcttcaaa agtcccacat cgcttagata agaaaacgaa gctgagttta
    15541 tatacagcta gagtcgaagt agtgattggg agaccaaccc agtggacata agcctgttcg
    15601 gttcgtaagc tgtaatgcaa gtagcgtatg cgctcacgca actggtccag aaccttgacc
    15661 gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt
    15721 ggggtacagt ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt
    15781 ttgatgttat ggagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta
    15841 aacatcatgg gggaagcggt gatcgccgaa gtatcgactc aactatcaga ggtagttggc
    15901 gtcatcgagc gccatctcga accgacgttg ctggccgtac atttgtacgg ctccgcagtg
    15961 gatggcggcc tgaagccaca cagtgatatt gatttgctgg ttacggtgac cgtaaggctt
    16021 gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg aaacttcggc ttcccctgga
    16081 gagagcgaga ttctccgcgc tgtagaagtc accattgttg tgcacgacga catcattccg
    16141 tggcgttatc cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa tgacattctt
    16201 gcaggtatct tcgagccagc cacgatcgac attgatctgg ctatcttgct gacaaaagca
    16261 agagaacata gcgttgcctt ggtaggtcca gcggcggagg aactctttga tccggttcct
    16321 gaacaggatc tatttgaggc gctaaatgaa accttaacgc tatggaactc gccgcccgac
    16381 tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc gcatttggta cagcgcagta
    16441 accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg caatggagcg cctgccggcc
    16501 cagtatcagc ccgtcatact tgaagctaga caggcttatc ttggacaaga agaagatcgc
    16561 ttggcctcgc gcgcagatca gttggaagaa tttgtccact acgtgaaagg cgagatcacc
    16621 aaggtagtcg gcaaataatg tctagctaga aattcgttca agccgacgcc gcttcgcggc
    16681 gcggcttaac tcaagcgtta gatgcactaa gcacataatt gctcacagcc aaactatcag
    16741 gtcaagtctg cttttattat ttttaagcgt gcataataag ccggtctcg
//
