LOCUS       sequence-390447-       16797 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cas9 fused with TLS2 mobility signal that confers graft 
            mobility. Induced by estradiol..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16797)
  AUTHORS   Yang L, Machin F, Wang S, Saplaoura E, Kragler F
  TITLE     Heritable transgene-free genome editing in plants by grafting of 
            wild-type shoots to transgenic donor rootstocks.
  JOURNAL   Nat Biotechnol. 2023 Jan 2. doi: 10.1038/s41587-022-01585-8.
  PUBMED    36593415
REFERENCE   2  (bases 1 to 16797)
  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: "Nat 
            Biotechnol. 2023 Jan 2. doi: 10.1038/s41587-022-01585-8."
COMMENT     Sequence Label: Cas9 TLS2
FEATURES             Location/Qualifiers
     source          1..16797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      869..1121
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        1444..1490
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             1524..1589
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1596..1616
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1641..5741
                     /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             1641..5741
                     /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             5654..5662
                     /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             5742..5789
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     protein_bind    6502..6526
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(6683..6707)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(7237..7254)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7237..7253)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7246..7268)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    7475..7499
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             8020..8811
                     /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"
     rep_origin      9057..9645
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9546..9565
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9799..9816
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    9831..9971
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(9897..9916)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     10057..10079
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      10315..10509
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(10575..11648)
                     /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(12077..12706)
                     /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    14037..14061
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     promoter        14252..14298
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             14327..14578
                     /codon_start=1
                     /gene="lexA (truncated)"
                     /product="DNA-binding domain of E. coli LexA"
                     /label=LexA BD
                     /note="binds the 16-bp palindromic sequence 
                     5'-CTGTATATATATACAG-3'"
                     /translation="MKALTARQQEVFDLIRDHISQTGMPPTRAEIAQRLGFRSPNAAEE
                     HLKALARKGVIEIVSGASRGIRLLQEEEEGLPLVGRVAA"
     CDS             14594..14827
                     /codon_start=1
                     /gene="UL48"
                     /product="transcriptional activation domain of herpes 
                     simplex virus protein VP16 (Triezenberg et al., 1988; 
                     Cousens et al., 1989)"
                     /label=VP16 AD
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     CDS             14837..15772
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPGKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             14885..15778
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPGKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATV"
     promoter        16336..16515
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        16368..16551
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
ORIGIN
        1 cactttgcat cggccgcgct cccgattccg gaagtgcttg acattgggga attcagcgag
       61 agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa
      121 accgaactgc ccgctgttct gcagccggtc gcggaggcca tggatgcgat cgctgcggcc
      181 gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg tcaatacact
      241 acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg gcaaactgtg
      301 atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat gctttgggcc
      361 gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa caatgtcctg
      421 acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt cggggattcc
      481 caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat ggagcagcag
      541 acgcgctact tcgagcggag gcatccggag cttgcaggat cgccgcggct ccgggcgtat
      601 atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa tttcgatgat
      661 gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg gactgtcggg
      721 cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt agaagtactc
      781 gccgatagtg gaaaccgacg ccccagcact cgtccgggat cttggaggtg atgtaacatg
      841 atcacaagct gatcccccga atttccccga tcgttcaaac atttggcaat aaagtttctt
      901 aagattgaat cctgttgccg gtcttgcgat gattatcata taatttctgt tgaattacgt
      961 taagcatgta ataattaaca tgtaatgcat gacgttattt atgagatggg tttttatgat
     1021 tagagtcccg caattataca tttaatacgc gatagaaaac aaaatatagc gcgcaaacta
     1081 ggataaatta tcgcgcgcgg tgtcatctat gttactagat cggggaattg atcccccctc
     1141 gacagcttgc atgccagctt gggctgcagg tcgaggctaa aaaactaatc gcattatcat
     1201 cccctcgacg tactgtacat ataaccactg gttttatata cagcagtact gtacatataa
     1261 ccactggttt tatatacagc agtcgacgta ctgtacatat aaccactggt tttatataca
     1321 gcagtactgt acatataacc actggtttta tatacagcag tcgaggtaag attagatatg
     1381 gatatgtata tggatatgta tatggtggta atgccatgta atatgctcga ctctaggatc
     1441 ttcgcaagac ccttcctcta tataaggaag ttcatttcat ttggagagga cacgctgaag
     1501 ctagtcgact ctagcctcga atggattaca aggaccacga cggggattac aaggaccacg
     1561 acattgatta caaggatgat gatgacaaga tggctccgaa gaagaagagg aaggttggca
     1621 tccacggggt gccagctgct gacaagaagt actcgatcgg cctcgatatt gggactaact
     1681 ctgttggctg ggccgtgatc accgacgagt acaaggtgcc ctcaaagaag ttcaaggtcc
     1741 tgggcaacac cgatcggcat tccatcaaga agaatctcat tggcgctctc ctgttcgaca
     1801 gcggcgagac ggctgaggct acgcggctca agcgcaccgc ccgcaggcgg tacacgcgca
     1861 ggaagaatcg catctgctac ctgcaggaga ttttctccaa cgagatggcg aaggttgacg
     1921 attctttctt ccacaggctg gaggagtcat tcctcgtgga ggaggataag aagcacgagc
     1981 ggcatccaat cttcggcaac attgtcgacg aggttgccta ccacgagaag taccctacga
     2041 tctaccatct gcggaagaag ctcgtggact ccacagataa ggcggacctc cgcctgatct
     2101 acctcgctct ggcccacatg attaagttca ggggccattt cctgatcgag ggggatctca
     2161 acccggacaa tagcgatgtt gacaagctgt tcatccagct cgtgcagacg tacaaccagc
     2221 tcttcgagga gaaccccatt aatgcgtcag gcgtcgacgc gaaggctatc ctgtccgcta
     2281 ggctctcgaa gtctcggcgc ctcgagaacc tgatcgccca gctgccgggc gagaagaaga
     2341 acggcctgtt cgggaatctc attgcgctca gcctggggct cacgcccaac ttcaagtcga
     2401 atttcgatct cgctgaggac gccaagctgc agctctccaa ggacacatac gacgatgacc
     2461 tggataacct cctggcccag atcggcgatc agtacgcgga cctgttcctc gctgccaaga
     2521 atctgtcgga cgccatcctc ctgtctgata ttctcagggt gaacaccgag attacgaagg
     2581 ctccgctctc agcctccatg atcaagcgct acgacgagca ccatcaggat ctgaccctcc
     2641 tgaaggcgct ggtcaggcag cagctccccg agaagtacaa ggagatcttc ttcgatcagt
     2701 cgaagaacgg ctacgctggg tacattgacg gcggggcctc tcaggaggag ttctacaagt
     2761 tcatcaagcc gattctggag aagatggacg gcacggagga gctgctggtg aagctcaatc
     2821 gcgaggacct cctgaggaag cagcggacat tcgataacgg cagcatccca caccagattc
     2881 atctcgggga gctgcacgct atcctgagga ggcaggagga cttctaccct ttcctcaagg
     2941 ataaccgcga gaagatcgag aagattctga ctttcaggat cccgtactac gtcggcccac
     3001 tcgctagggg caactcccgc ttcgcttgga tgacccgcaa gtcagaggag acgatcacgc
     3061 cgtggaactt cgaggaggtg gtcgacaagg gcgctagcgc tcagtcgttc atcgagagga
     3121 tgacgaattt cgacaagaac ctgccaaatg agaaggtgct ccctaagcac tcgctcctgt
     3181 acgagtactt cacagtctac aacgagctga ctaaggtgaa gtatgtgacc gagggcatga
     3241 ggaagccggc tttcctgtct ggggagcaga agaaggccat cgtggacctc ctgttcaaga
     3301 ccaaccggaa ggtcacggtt aagcagctca aggaggacta cttcaagaag attgagtgct
     3361 tcgattcggt cgagatctct ggcgttgagg accgcttcaa cgcctccctg gggacctacc
     3421 acgatctcct gaagatcatt aaggataagg acttcctgga caacgaggag aatgaggata
     3481 tcctcgagga cattgtgctg acactcactc tgttcgagga ccgggagatg atcgaggagc
     3541 gcctgaagac ttacgcccat ctcttcgatg acaaggtcat gaagcagctc aagaggagga
     3601 ggtacaccgg ctgggggagg ctgagcagga agctcatcaa cggcattcgg gacaagcagt
     3661 ccgggaagac gatcctcgac ttcctgaaga gcgatggctt cgcgaaccgc aatttcatgc
     3721 agctgattca cgatgacagc ctcacattca aggaggatat ccagaaggct caggtgagcg
     3781 gccaggggga ctcgctgcac gagcatatcg cgaacctcgc tggctcgcca gctatcaaga
     3841 aggggattct gcagaccgtg aaggttgtgg acgagctggt gaaggtcatg ggcaggcaca
     3901 agcctgagaa catcgtcatt gagatggccc gggagaatca gaccacgcag aagggccaga
     3961 agaactcacg cgagaggatg aagaggatcg aggagggcat taaggagctg gggtcccaga
     4021 tcctcaagga gcacccggtg gagaacacgc agctgcagaa tgagaagctc tacctgtact
     4081 acctccagaa tggccgcgat atgtatgtgg accaggagct ggatattaac aggctcagcg
     4141 attacgacgt cgatcatatc gttccacagt cattcctgaa ggatgactcc attgacaaca
     4201 aggtcctcac caggtcggac aagaaccggg gcaagtctga taatgttcct tcagaggagg
     4261 tcgttaagaa gatgaagaac tactggcgcc agctcctgaa tgccaagctg atcacgcagc
     4321 ggaagttcga taacctcaca aaggctgaga ggggcgggct ctctgagctg gacaaggcgg
     4381 gcttcatcaa gaggcagctg gtcgagacac ggcagatcac taagcacgtt gcgcagattc
     4441 tcgactcacg gatgaacact aagtacgatg agaatgacaa gctgatccgc gaggtgaagg
     4501 tcatcaccct gaagtcaaag ctcgtctccg acttcaggaa ggatttccag ttctacaagg
     4561 ttcgggagat caacaattac caccatgccc atgacgcgta cctgaacgcg gtggtcggca
     4621 cagctctgat caagaagtac ccaaagctcg agagcgagtt cgtgtacggg gactacaagg
     4681 tttacgatgt gaggaagatg atcgccaagt cggagcagga gattggcaag gctaccgcca
     4741 agtacttctt ctactctaac attatgaatt tcttcaagac agagatcact ctggccaatg
     4801 gcgagatccg gaagcgcccc ctcatcgaga cgaacggcga gacgggggag atcgtgtggg
     4861 acaagggcag ggatttcgcg accgtcagga aggttctctc catgccacaa gtgaatatcg
     4921 tcaagaagac agaggtccag actggcgggt tctctaagga gtcaattctg cctaagcgga
     4981 acagcgacaa gctcatcgcc cgcaagaagg actgggatcc gaagaagtac ggcgggttcg
     5041 acagccccac tgtggcctac tcggtcctgg ttgtggcgaa ggttgagaag ggcaagtcca
     5101 agaagctcaa gagcgtgaag gagctgctgg ggatcacgat tatggagcgc tccagcttcg
     5161 agaagaaccc gatcgatttc ctggaggcga agggctacaa ggaggtgaag aaggacctga
     5221 tcattaagct ccccaagtac tcactcttcg agctggagaa cggcaggaag cggatgctgg
     5281 cttccgctgg cgagctgcag aaggggaacg agctggctct gccgtccaag tatgtgaact
     5341 tcctctacct ggcctcccac tacgagaagc tcaagggcag ccccgaggac aacgagcaga
     5401 agcagctgtt cgtcgagcag cacaagcatt acctcgacga gatcattgag cagatttccg
     5461 agttctccaa gcgcgtgatc ctggccgacg cgaatctgga taaggtcctc tccgcgtaca
     5521 acaagcaccg cgacaagcca atcagggagc aggctgagaa tatcattcat ctcttcaccc
     5581 tgacgaacct cggcgcccct gctgctttca agtacttcga cacaactatc gatcgcaaga
     5641 ggtacacaag cactaaggag gtcctggacg cgaccctcat ccaccagtcg attaccggcc
     5701 tctacgagac gcgcatcgac ctgtctcagc tcgggggcga caagcggcca gcggcgacga
     5761 agaaggcggg gcaggcgaag aagaagaagt gaattatcag agtggtggtg ggcccataac
     5821 ccacaggtcc gctctgatat cgagagagct ttcgttcgta tcatcggttt cgacaacgtt
     5881 cgtcaagttc aatgcatcag tttcattgcg cacacaccag aatcctactg agtttgagta
     5941 ttatggcatt gggaaaactg tttttcttgt accatttgtt gtgcttgtaa tttactgtgt
     6001 tttttattcg gttttcgcta tcgaactgtg aaatggaaat ggatggagaa gagttaatga
     6061 atgatatggt ccttttgttc attctcaaat taatattatt tgttttttct cttatttgtt
     6121 gtgtgttgaa tttgaaatta taagagatat gcaaacattt tgttttgagt aaaaatgtgt
     6181 caaatcgtgg cctctaatga ccgaagttaa tatgaggagt aaaacacttg tagttgtacc
     6241 attatgctta ttcactaggc aacaaatata ttttcagacc tagaaaagct gcaaatgtta
     6301 ctgaatacaa gtatgtcctc ttgtgtttta gacatttatg aactttcctt tatgtaattt
     6361 tccagaatcc ttgtcagatt ctaatcattg ctttataatt atagttatac tcatggattt
     6421 gtagttgagt atgaaaatat tttttaatgc attttatgac ttgccaattg attgacaacg
     6481 tcgaggcgcg ccaagctatc aacaagtttg tacaaaaaag caggctccac catgggaacc
     6541 aattcagtcg actggatccg gtaccgaatt cgcttactaa aagccagata acagtatgcg
     6601 tatttgcgcg ctgatttttg cggtataaga atatatactg atatgtatac ccgaagtatg
     6661 tcaaaaagag gtgtgcttct agacccagct ttcttgtaca aagtggttga taattcttaa
     6721 ttaactagtc gatccaggcc tcccagcttt cgtccgtatc atcggtttcg acaacgttcg
     6781 tcaagttcaa tgcatcagtt tcattgccca cacaccagaa tcctactaag tttgagtatt
     6841 atggcattgg aaaagctgtt ttcttctatc atttgttctg cttgtaattt actgtgttct
     6901 ttcagttttt gttttcggac atcaaaatgc aaatggatgg ataagagtta ataaatgata
     6961 tggtcctttt gttcattctc aaattattat tatctgttgt ttttacttta atgggttgaa
     7021 tttaagtaag aaaggaacta acagtgtgat attaaggtgc aatgttagac atataaaaca
     7081 gtctttcacc tctctttggt tatgtcttga attggtttgt ttcttcactt atctgtgtaa
     7141 tcaagtttac tatgagtcta tgatcaagta attatgcaat caagttaagt acagtatagg
     7201 ctttttgtgt cgagggggta ccgagtcgag gaattcactg gccgtcgttt tacaacgtcg
     7261 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     7321 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     7381 gaatggcggg taccgagctc gaattcaatt cggcgttaat tcagtacatt aaaaacgtcc
     7441 gcaatgtgtt attaagttgt ctaagcgtca atttgtttac accacaatat atcctgccac
     7501 cagccagcca acagctcccc gaccggcagc tcggcacaaa atcaccactc gatacaggca
     7561 gcccatcagt ccgggacggc gtcagcggga gagccgttgt aaggcggcag actttgctca
     7621 tgttaccgat gctattcgga agaacggcaa ctaagctgcc gggtttgaaa cacggatgat
     7681 ctcgcggagg gtagcatgtt gattgtaacg atgacagagc gttgctgcct gtgatcaatt
     7741 cgggcacgaa cccagtggac ataagcctgt tcggttcgta agctgtaatg caagtagcgt
     7801 atgcgctcac gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg
     7861 gcggttttca tggcttgtta tgactgtttt tttggggtac agtctatgcc tcgggcatcc
     7921 aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt
     7981 acgcagcagg gcagtcgccc taaaacaaag ttaaacatca tgggggaagc ggtgatcgcc
     8041 gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg
     8101 ttgctggccg tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagtgat
     8161 attgatttgc tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc
     8221 aacgaccttt tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa
     8281 gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg
     8341 caatttggag aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc
     8401 gacattgatc tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt
     8461 ccagcggcgg aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat
     8521 gaaaccttaa cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg
     8581 cttacgttgt cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc
     8641 gctgccgact gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct
     8701 agacaggctt atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa
     8761 gaatttgtcc actacgtgaa aggcgagatc accaaggtag tcggcaaata atgtctagct
     8821 agaaattcgt tcaagccgac gccgcttcgc ggcgcggctt aactcaagcg ttagatgcac
     8881 taagcacata attgctcaca gccaaactat caggtcaagt ctgcttttat tatttttaag
     8941 cgtgcataat aagccctaca caaattggga gatatatcat gcatgaccaa aatcccttaa
     9001 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     9061 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     9121 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     9181 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     9241 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     9301 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     9361 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     9421 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     9481 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     9541 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     9601 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     9661 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     9721 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     9781 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     9841 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca
     9901 atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg
     9961 tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
    10021 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
    10081 tttcaccgtc atcaccgaaa cgcgcgaggc agggtgcctt gatgtgggcg ccggcggtcg
    10141 agtggcgacg gcgcggcttg tccgcgccct ggtagattgc ctggccgtag gccagccatt
    10201 tttgagcggc cagcggccgc gataggccga cgcgaagcgg cggggcgtag ggagcgcagc
    10261 gaccgaaggg taggcgcttt ttgcagctct tcggctgtgc gctggccaga cagttatgca
    10321 caggccaggc gggttttaag agttttaata agttttaaag agttttaggc ggaaaaatcg
    10381 ccttttttct cttttatatc agtcacttac atgtgtgacc ggttcccaat gtacggcttt
    10441 gggttcccaa tgtacgggtt ccggttccca atgtacggct ttgggttccc aatgtacgtg
    10501 ctatccacag gaaagagacc ttttcgacct ttttcccctg ctagggcaat ttgccctagc
    10561 atctgctccg tacattagga accggcggat gcttcgccct cgatcaggtt gcggtagcgc
    10621 atgactagga tcgggccagc ctgccccgcc tcctccttca aatcgtactc cggcaggtca
    10681 tttgacccga tcagcttgcg cacggtgaaa cagaacttct tgaactctcc ggcgctgcca
    10741 ctgcgttcgt agatcgtctt gaacaaccat ctggcttctg ccttgcctgc ggcgcggcgt
    10801 gccaggcggt agagaaaacg gccgatgccg ggatcgatca aaaagtaatc ggggtgaacc
    10861 gtcagcacgt ccgggttctt gccttctgtg atctcgcggt acatccaatc agctagctcg
    10921 atctcgatgt actccggccg cccggtttcg ctctttacga tcttgtagcg gctaatcaag
    10981 gcttcaccct cggataccgt caccaggcgg ccgttcttgg ccttcttcgt acgctgcatg
    11041 gcaacgtgcg tggtgtttaa ccgaatgcag gtttctacca ggtcgtcttt ctgctttccg
    11101 ccatcggctc gccggcagaa cttgagtacg tccgcaacgt gtggacggaa cacgcggccg
    11161 ggcttgtctc ccttcccttc ccggtatcgg ttcatggatt cggttagatg ggaaaccgcc
    11221 atcagtacca ggtcgtaatc ccacacactg gccatgccgg ccggccctgc ggaaacctct
    11281 acgtgcccgt ctggaagctc gtagcggatc acctcgccag ctcgtcggtc acgcttcgac
    11341 agacggaaaa cggccacgtc catgatgctg cgactatcgc gggtgcccac gtcatagagc
    11401 atcggaacga aaaaatctgg ttgctcgtcg cccttgggcg gcttcctaat cgacggcgca
    11461 ccggctgccg gcggttgccg ggattctttg cggattcgat cagcggccgc ttgccacgat
    11521 tcaccggggc gtgcttctgc ctcgatgcgt tgccgctggg cggcctgcgc ggccttcaac
    11581 ttctccacca ggtcatcacc cagcgccgcg ccgatttgta ccgggccgga tggtttgcga
    11641 ccgctcacgc cgattcctcg ggcttggggg ttccagtgcc attgcagggc cggcagacaa
    11701 cccagccgct tacgcctggc caaccgcccg ttcctccaca catggggcat tccacggcgt
    11761 cggtgcctgg ttgttcttga ttttccatgc cgcctccttt agccgctaaa attcatctac
    11821 tcatttattc atttgctcat ttactctggt agctgcgcga tgtattcaga tagcagctcg
    11881 gtaatggtct tgccttggcg taccgcgtac atcttcagct tggtgtgatc ctccgccggc
    11941 aactgaaagt tgacccgctt catggctggc gtgtctgcca ggctggccaa cgttgcagcc
    12001 ttgctgctgc gtgcgctcgg acggccggca cttagcgtgt ttgtgctttt gctcattttc
    12061 tctttacctc attaactcaa atgagttttg atttaatttc agcggccagc gcctggacct
    12121 cgcgggcagc gtcgccctcg ggttctgatt caagaacggt tgtgccggcg gcggcagtgc
    12181 ctgggtagct cacgcgctgc gtgatacggg actcaagaat gggcagctcg tacccggcca
    12241 gcgcctcggc aacctcaccg ccgatgcgcg tgcctttgat cgcccgcgac acgacaaagg
    12301 ccgcttgtag ccttccatcc gtgacctcaa tgcgctgctt aaccagctcc accaggtcgg
    12361 cggtggccca tatgtcgtaa gggcttggct gcaccggaat cagcacgaag tcggctgcct
    12421 tgatcgcgga cacagccaag tccgccgcct ggggcgctcc gtcgatcact acgaagtcgc
    12481 gccggccgat ggccttcacg tcgcggtcaa tcgtcgggcg gtcgatgccg acaacggtta
    12541 gcggttgatc ttcccgcacg gccgcccaat cgcgggcact gccctgggga tcggaatcga
    12601 ctaacagaac atcggccccg gcgagttgca gggcgcgggc tagatgggtt gcgatggtcg
    12661 tcttgcctga cccgcctttc tggttaagta cagcgataac cttcatgcgt tccccttgcg
    12721 tatttgttta tttactcatc gcatcatata cgcagcgacc gcatgacgca agctgtttta
    12781 ctcaaataca catcaccttt ttagacggcg gcgctcggtt tcttcagcgg ccaagctggc
    12841 cggccaggcc gccagcttgg catcagacaa accggccagg atttcatgca gccgcacggt
    12901 tgagacgtgc gcgggcggct cgaacacgta cccggccgcg atcatctccg cctcgatctc
    12961 ttcggtaatg aaaaacggtt cgtcctggcc gtcctggtgc ggtttcatgc ttgttcctct
    13021 tggcgttcat tctcggcggc cgccagggcg tcggcctcgg tcaatgcgtc ctcacggaag
    13081 gcaccgcgcc gcctggcctc ggtgggcgtc acttcctcgc tgcgctcaag tgcgcggtac
    13141 agggtcgagc gatgcacgcc aagcagtgca gccgcctctt tcacggtgcg gccttcctgg
    13201 tcgatcagct cgcgggcgtg cgcgatctgt gccggggtga gggtagggcg ggggccaaac
    13261 ttcacgcctc gggccttggc ggcctcgcgc ccgctccggg tgcggtcgat gattagggaa
    13321 cgctcgaact cggcaatgcc ggcgaacacg gtcaacacca tgcggccggc cggcgtggtg
    13381 gtgtcggccc acggctctgc caggctacgc aggcccgcgc cggcctcctg gatgcgctcg
    13441 gcaatgtcca gtaggtcgcg ggtgctgcgg gccaggcggt ctagcctggt cactgtcaca
    13501 acgtcgccag ggcgtaggtg gtcaagcatc ctggccagct ccgggcggtc gcgcctggtg
    13561 ccggtgatct tctcggaaaa cagcttggtg cagccggccg cgtgcagttc ggcccgttgg
    13621 ttggtcaagt cctggtcgtc ggtgctgacg cgggcatagc ccagcaggcc agcggcggcg
    13681 ctcttgttca tggcgtaatg tctccggttc tagtcgcaag tattctactt tatgcgacta
    13741 aaacacgcga caagaaaacg ccaggaaaag ggcagggcgg cagcctgtcg cgtaacttag
    13801 gacttgtgcg acatgtcgtt ttcagaagac ggctgcactg aacgtcagaa gccgactgca
    13861 ctatagcagc ggaggggttg gatcaaagta ctttaaagta ctttaaagta ctttaaagta
    13921 ctttgatccc gaggggaacc ctgtggttgg catgcacata caaatggacg aacggataaa
    13981 ccttttcacg cccttttaaa tatccgatta ttctaataaa cgctcttttc tcttaggttt
    14041 acccgccaat atatcctgtc aaacactgat agtttaaact gaaggcggga aacgacaatc
    14101 tgatccaagc tcaagctaag cttgcatgcc tgcaggatat cgtggatcca agcttgccac
    14161 gtgccgccac gtgccgccac gtgccgccac gtgcctctag aggatccatc tccactgacg
    14221 taagggatga cgcacaatcc cactatcctt cgcaagaccc ttcctctata taaggaagtt
    14281 catttcattt ggagaggaca cgctgggatc cccaattccg ggcggaatga aagcgttaac
    14341 ggccaggcaa caagaggtgt ttgatctcat ccgtgatcac atcagccaga caggtatgcc
    14401 gccgacgcgt gcggaaatcg cgcagcgttt ggggttccgt tccccaaacg cggctgaaga
    14461 acatctgaag gcgctggcac gcaaaggcgt tattgaaatt gtttccggcg catcacgcgg
    14521 gattcgtctg ttgcaggaag aggaagaagg gttgccgctg gtaggtcgtg tggctgccgg
    14581 tgaaccgtcg agcgcccccc cgaccgatgt cagcctgggg gacgagctcc acttagacgg
    14641 cgaggacgtg gcgatggcgc atgccgacgc gctagacgat ttcgatctgg acatgttggg
    14701 ggacggggat tccccgggtc cgggatttac cccccacgac tccgccccct acggcgctct
    14761 ggatatggcc gacttcgagt ttgagcagat gtttaccgat gcccttggaa ttgacgagta
    14821 cggtggggat ccgtctgctg gagacatgag agctgccaac ctttggccaa gcccgctcat
    14881 gatcaaacgc tctaagaaga acagcctggc cttgtccctg acggccgacc agatggtcag
    14941 tgccttgttg gatgctgagc cccccatact ctattccgag tatgatccta ccagaccctt
    15001 cagtgaagct tcgatgatgg gcttactgac caacctggca gacagggagc tggttcacat
    15061 gatcaactgg gcgaagaggg tgccaggctt tgtggatttg accctccatg atcaggtcca
    15121 ccttctagaa tgtgcctggc tagagatcct gatgattggt ctcgtctggc gctccatgga
    15181 gcacccaggg aagctactgt ttgctcctaa cttgctcttg gacaggaacc agggaaaatg
    15241 tgtagagggc atggtggaga tcttcgacat gctgctggct acatcatctc ggttccgcat
    15301 gatgaatctg cagggagagg agtttgtgtg cctcaaatct attattttgc ttaattctgg
    15361 agtgtacaca tttctgtcca gcaccctgaa gtctctggaa gagaaggacc atatccaccg
    15421 agtcctggac aagatcacag acactttgat ccacctgatg gccaaggcag gcctgaccct
    15481 gcagcagcag caccagcggc tggcccagct cctcctcatc ctctcccaca tcaggcacat
    15541 gagtaacaaa ggcatggagc atctgtacag catgaagtgc aagaacgtgg tgcccctcta
    15601 tgacctgctg ctggagatgc tggacgccca ccgcctacat gcgcccacta gccgtggagg
    15661 ggcatccgtg gaggagacgg accaaagcca cttggccact gcgggctcta cttcatcgca
    15721 ttccttgcaa aagtattaca tcacggggga ggcagagggt ttccctgcca cagtctgaga
    15781 gctccctggc ggaattccca gagatgttag ctgaaatcat cactaatcag ataccaaaat
    15841 attcaaatgg aaatatcaaa aagcttctgt ttcatcaaaa atgactcgac ctaactgagt
    15901 aagctagctt gttcgagtat tatggcattg ggaaaactgt ttttcttgta ccatttgttg
    15961 tgcttgtaat ttactgtgtt ttttattcgg ttttcgctat cgaactgtga aatggaaatg
    16021 gatggagaag agttaatgaa tgatatggtc cttttgttca ttctcaaatt aatattattt
    16081 gttttttctc ttatttgttg tgtgttgaat ttgaaattat aagagatatg caaacatttt
    16141 gttttgagta aaaatgtgtc aaatcgtggc ctctaatgac cgaagttaat atgaggagta
    16201 aaacactaga tccccaaaca agcttgaaac tgaaggcggg aaacgacaat ctgatcatga
    16261 gcggagaatt aagggagtca cgttatgacc cccgccgatg acgcgggaca agccgtttta
    16321 cgtttggaac tgacagaacc gcaacgttga aggagccact cagccgcggg tttctggagt
    16381 ttaatgagct aagcacatac gtcagaaacc attattgcgc gttcaaaagt cgcctaaggt
    16441 cactatcagc tagcaaatat ttcttgtcaa aaatgctcca ctgacgttcc ataaattccc
    16501 ctcggtatcc aattagagtc tcatattcac tctcaatcca aataatctgc accggatccg
    16561 ctagaggatc tcgacctgca agatcccggg gggcaatgag atatgaaaaa gcctgaactc
    16621 accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc cgacctgatg
    16681 cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg gcgtggatat
    16741 gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt ttatcgg
//
