LOCUS       sequence-390448-       16750 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cas9 without a mobility signal and the mRNA is not graft 
            mobile. Induced by estradiol..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16750)
  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 16750)
  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 Null
FEATURES             Location/Qualifiers
     source          1..16750
                     /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    6455..6479
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(6636..6660)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(7190..7207)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7190..7206)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7199..7221)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    7428..7452
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             7973..8764
                     /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      9010..9598
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9499..9518
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9752..9769
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    9784..9924
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(9850..9869)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     10010..10032
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      10268..10462
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(10528..11601)
                     /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(12030..12659)
                     /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    13990..14014
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     promoter        14205..14251
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             14280..14531
                     /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             14547..14780
                     /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             14790..15725
                     /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             14838..15731
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPGKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATV"
     promoter        16289..16468
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        16321..16504
                     /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 gatcgagaga gctttcgttc gtatcatcgg
     5821 tttcgacaac gttcgtcaag ttcaatgcat cagtttcatt gcgcacacac cagaatccta
     5881 ctgagtttga gtattatggc attgggaaaa ctgtttttct tgtaccattt gttgtgcttg
     5941 taatttactg tgttttttat tcggttttcg ctatcgaact gtgaaatgga aatggatgga
     6001 gaagagttaa tgaatgatat ggtccttttg ttcattctca aattaatatt atttgttttt
     6061 tctcttattt gttgtgtgtt gaatttgaaa ttataagaga tatgcaaaca ttttgttttg
     6121 agtaaaaatg tgtcaaatcg tggcctctaa tgaccgaagt taatatgagg agtaaaacac
     6181 ttgtagttgt accattatgc ttattcacta ggcaacaaat atattttcag acctagaaaa
     6241 gctgcaaatg ttactgaata caagtatgtc ctcttgtgtt ttagacattt atgaactttc
     6301 ctttatgtaa ttttccagaa tccttgtcag attctaatca ttgctttata attatagtta
     6361 tactcatgga tttgtagttg agtatgaaaa tattttttaa tgcattttat gacttgccaa
     6421 ttgattgaca acgtcgaggc gcgccaagct atcaacaagt ttgtacaaaa aagcaggctc
     6481 caccatggga accaattcag tcgactggat ccggtaccga attcgcttac taaaagccag
     6541 ataacagtat gcgtatttgc gcgctgattt ttgcggtata agaatatata ctgatatgta
     6601 tacccgaagt atgtcaaaaa gaggtgtgct tctagaccca gctttcttgt acaaagtggt
     6661 tgataattct taattaacta gtcgatccag gcctcccagc tttcgtccgt atcatcggtt
     6721 tcgacaacgt tcgtcaagtt caatgcatca gtttcattgc ccacacacca gaatcctact
     6781 aagtttgagt attatggcat tggaaaagct gttttcttct atcatttgtt ctgcttgtaa
     6841 tttactgtgt tctttcagtt tttgttttcg gacatcaaaa tgcaaatgga tggataagag
     6901 ttaataaatg atatggtcct tttgttcatt ctcaaattat tattatctgt tgtttttact
     6961 ttaatgggtt gaatttaagt aagaaaggaa ctaacagtgt gatattaagg tgcaatgtta
     7021 gacatataaa acagtctttc acctctcttt ggttatgtct tgaattggtt tgtttcttca
     7081 cttatctgtg taatcaagtt tactatgagt ctatgatcaa gtaattatgc aatcaagtta
     7141 agtacagtat aggctttttg tgtcgagggg gtaccgagtc gaggaattca ctggccgtcg
     7201 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     7261 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     7321 agttgcgcag cctgaatggc gggtaccgag ctcgaattca attcggcgtt aattcagtac
     7381 attaaaaacg tccgcaatgt gttattaagt tgtctaagcg tcaatttgtt tacaccacaa
     7441 tatatcctgc caccagccag ccaacagctc cccgaccggc agctcggcac aaaatcacca
     7501 ctcgatacag gcagcccatc agtccgggac ggcgtcagcg ggagagccgt tgtaaggcgg
     7561 cagactttgc tcatgttacc gatgctattc ggaagaacgg caactaagct gccgggtttg
     7621 aaacacggat gatctcgcgg agggtagcat gttgattgta acgatgacag agcgttgctg
     7681 cctgtgatca attcgggcac gaacccagtg gacataagcc tgttcggttc gtaagctgta
     7741 atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg
     7801 taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgggg tacagtctat
     7861 gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag
     7921 cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca tcatggggga
     7981 agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca
     8041 tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa
     8101 gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg
     8161 gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct
     8221 ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc
     8281 taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga
     8341 gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt
     8401 tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt
     8461 tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga
     8521 gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc
     8581 gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt
     8641 catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc
     8701 agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg tagtcggcaa
     8761 ataatgtcta gctagaaatt cgttcaagcc gacgccgctt cgcggcgcgg cttaactcaa
     8821 gcgttagatg cactaagcac ataattgctc acagccaaac tatcaggtca agtctgcttt
     8881 tattattttt aagcgtgcat aataagccct acacaaattg ggagatatat catgcatgac
     8941 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     9001 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     9061 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     9121 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     9181 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     9241 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     9301 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     9361 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     9421 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     9481 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     9541 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     9601 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     9661 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     9721 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     9781 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg
     9841 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
     9901 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
     9961 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
    10021 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagggtgc cttgatgtgg
    10081 gcgccggcgg tcgagtggcg acggcgcggc ttgtccgcgc cctggtagat tgcctggccg
    10141 taggccagcc atttttgagc ggccagcggc cgcgataggc cgacgcgaag cggcggggcg
    10201 tagggagcgc agcgaccgaa gggtaggcgc tttttgcagc tcttcggctg tgcgctggcc
    10261 agacagttat gcacaggcca ggcgggtttt aagagtttta ataagtttta aagagtttta
    10321 ggcggaaaaa tcgccttttt tctcttttat atcagtcact tacatgtgtg accggttccc
    10381 aatgtacggc tttgggttcc caatgtacgg gttccggttc ccaatgtacg gctttgggtt
    10441 cccaatgtac gtgctatcca caggaaagag accttttcga cctttttccc ctgctagggc
    10501 aatttgccct agcatctgct ccgtacatta ggaaccggcg gatgcttcgc cctcgatcag
    10561 gttgcggtag cgcatgacta ggatcgggcc agcctgcccc gcctcctcct tcaaatcgta
    10621 ctccggcagg tcatttgacc cgatcagctt gcgcacggtg aaacagaact tcttgaactc
    10681 tccggcgctg ccactgcgtt cgtagatcgt cttgaacaac catctggctt ctgccttgcc
    10741 tgcggcgcgg cgtgccaggc ggtagagaaa acggccgatg ccgggatcga tcaaaaagta
    10801 atcggggtga accgtcagca cgtccgggtt cttgccttct gtgatctcgc ggtacatcca
    10861 atcagctagc tcgatctcga tgtactccgg ccgcccggtt tcgctcttta cgatcttgta
    10921 gcggctaatc aaggcttcac cctcggatac cgtcaccagg cggccgttct tggccttctt
    10981 cgtacgctgc atggcaacgt gcgtggtgtt taaccgaatg caggtttcta ccaggtcgtc
    11041 tttctgcttt ccgccatcgg ctcgccggca gaacttgagt acgtccgcaa cgtgtggacg
    11101 gaacacgcgg ccgggcttgt ctcccttccc ttcccggtat cggttcatgg attcggttag
    11161 atgggaaacc gccatcagta ccaggtcgta atcccacaca ctggccatgc cggccggccc
    11221 tgcggaaacc tctacgtgcc cgtctggaag ctcgtagcgg atcacctcgc cagctcgtcg
    11281 gtcacgcttc gacagacgga aaacggccac gtccatgatg ctgcgactat cgcgggtgcc
    11341 cacgtcatag agcatcggaa cgaaaaaatc tggttgctcg tcgcccttgg gcggcttcct
    11401 aatcgacggc gcaccggctg ccggcggttg ccgggattct ttgcggattc gatcagcggc
    11461 cgcttgccac gattcaccgg ggcgtgcttc tgcctcgatg cgttgccgct gggcggcctg
    11521 cgcggccttc aacttctcca ccaggtcatc acccagcgcc gcgccgattt gtaccgggcc
    11581 ggatggtttg cgaccgctca cgccgattcc tcgggcttgg gggttccagt gccattgcag
    11641 ggccggcaga caacccagcc gcttacgcct ggccaaccgc ccgttcctcc acacatgggg
    11701 cattccacgg cgtcggtgcc tggttgttct tgattttcca tgccgcctcc tttagccgct
    11761 aaaattcatc tactcattta ttcatttgct catttactct ggtagctgcg cgatgtattc
    11821 agatagcagc tcggtaatgg tcttgccttg gcgtaccgcg tacatcttca gcttggtgtg
    11881 atcctccgcc ggcaactgaa agttgacccg cttcatggct ggcgtgtctg ccaggctggc
    11941 caacgttgca gccttgctgc tgcgtgcgct cggacggccg gcacttagcg tgtttgtgct
    12001 tttgctcatt ttctctttac ctcattaact caaatgagtt ttgatttaat ttcagcggcc
    12061 agcgcctgga cctcgcgggc agcgtcgccc tcgggttctg attcaagaac ggttgtgccg
    12121 gcggcggcag tgcctgggta gctcacgcgc tgcgtgatac gggactcaag aatgggcagc
    12181 tcgtacccgg ccagcgcctc ggcaacctca ccgccgatgc gcgtgccttt gatcgcccgc
    12241 gacacgacaa aggccgcttg tagccttcca tccgtgacct caatgcgctg cttaaccagc
    12301 tccaccaggt cggcggtggc ccatatgtcg taagggcttg gctgcaccgg aatcagcacg
    12361 aagtcggctg ccttgatcgc ggacacagcc aagtccgccg cctggggcgc tccgtcgatc
    12421 actacgaagt cgcgccggcc gatggccttc acgtcgcggt caatcgtcgg gcggtcgatg
    12481 ccgacaacgg ttagcggttg atcttcccgc acggccgccc aatcgcgggc actgccctgg
    12541 ggatcggaat cgactaacag aacatcggcc ccggcgagtt gcagggcgcg ggctagatgg
    12601 gttgcgatgg tcgtcttgcc tgacccgcct ttctggttaa gtacagcgat aaccttcatg
    12661 cgttcccctt gcgtatttgt ttatttactc atcgcatcat atacgcagcg accgcatgac
    12721 gcaagctgtt ttactcaaat acacatcacc tttttagacg gcggcgctcg gtttcttcag
    12781 cggccaagct ggccggccag gccgccagct tggcatcaga caaaccggcc aggatttcat
    12841 gcagccgcac ggttgagacg tgcgcgggcg gctcgaacac gtacccggcc gcgatcatct
    12901 ccgcctcgat ctcttcggta atgaaaaacg gttcgtcctg gccgtcctgg tgcggtttca
    12961 tgcttgttcc tcttggcgtt cattctcggc ggccgccagg gcgtcggcct cggtcaatgc
    13021 gtcctcacgg aaggcaccgc gccgcctggc ctcggtgggc gtcacttcct cgctgcgctc
    13081 aagtgcgcgg tacagggtcg agcgatgcac gccaagcagt gcagccgcct ctttcacggt
    13141 gcggccttcc tggtcgatca gctcgcgggc gtgcgcgatc tgtgccgggg tgagggtagg
    13201 gcgggggcca aacttcacgc ctcgggcctt ggcggcctcg cgcccgctcc gggtgcggtc
    13261 gatgattagg gaacgctcga actcggcaat gccggcgaac acggtcaaca ccatgcggcc
    13321 ggccggcgtg gtggtgtcgg cccacggctc tgccaggcta cgcaggcccg cgccggcctc
    13381 ctggatgcgc tcggcaatgt ccagtaggtc gcgggtgctg cgggccaggc ggtctagcct
    13441 ggtcactgtc acaacgtcgc cagggcgtag gtggtcaagc atcctggcca gctccgggcg
    13501 gtcgcgcctg gtgccggtga tcttctcgga aaacagcttg gtgcagccgg ccgcgtgcag
    13561 ttcggcccgt tggttggtca agtcctggtc gtcggtgctg acgcgggcat agcccagcag
    13621 gccagcggcg gcgctcttgt tcatggcgta atgtctccgg ttctagtcgc aagtattcta
    13681 ctttatgcga ctaaaacacg cgacaagaaa acgccaggaa aagggcaggg cggcagcctg
    13741 tcgcgtaact taggacttgt gcgacatgtc gttttcagaa gacggctgca ctgaacgtca
    13801 gaagccgact gcactatagc agcggagggg ttggatcaaa gtactttaaa gtactttaaa
    13861 gtactttaaa gtactttgat cccgagggga accctgtggt tggcatgcac atacaaatgg
    13921 acgaacggat aaaccttttc acgccctttt aaatatccga ttattctaat aaacgctctt
    13981 ttctcttagg tttacccgcc aatatatcct gtcaaacact gatagtttaa actgaaggcg
    14041 ggaaacgaca atctgatcca agctcaagct aagcttgcat gcctgcagga tatcgtggat
    14101 ccaagcttgc cacgtgccgc cacgtgccgc cacgtgccgc cacgtgcctc tagaggatcc
    14161 atctccactg acgtaaggga tgacgcacaa tcccactatc cttcgcaaga cccttcctct
    14221 atataaggaa gttcatttca tttggagagg acacgctggg atccccaatt ccgggcggaa
    14281 tgaaagcgtt aacggccagg caacaagagg tgtttgatct catccgtgat cacatcagcc
    14341 agacaggtat gccgccgacg cgtgcggaaa tcgcgcagcg tttggggttc cgttccccaa
    14401 acgcggctga agaacatctg aaggcgctgg cacgcaaagg cgttattgaa attgtttccg
    14461 gcgcatcacg cgggattcgt ctgttgcagg aagaggaaga agggttgccg ctggtaggtc
    14521 gtgtggctgc cggtgaaccg tcgagcgccc ccccgaccga tgtcagcctg ggggacgagc
    14581 tccacttaga cggcgaggac gtggcgatgg cgcatgccga cgcgctagac gatttcgatc
    14641 tggacatgtt gggggacggg gattccccgg gtccgggatt taccccccac gactccgccc
    14701 cctacggcgc tctggatatg gccgacttcg agtttgagca gatgtttacc gatgcccttg
    14761 gaattgacga gtacggtggg gatccgtctg ctggagacat gagagctgcc aacctttggc
    14821 caagcccgct catgatcaaa cgctctaaga agaacagcct ggccttgtcc ctgacggccg
    14881 accagatggt cagtgccttg ttggatgctg agccccccat actctattcc gagtatgatc
    14941 ctaccagacc cttcagtgaa gcttcgatga tgggcttact gaccaacctg gcagacaggg
    15001 agctggttca catgatcaac tgggcgaaga gggtgccagg ctttgtggat ttgaccctcc
    15061 atgatcaggt ccaccttcta gaatgtgcct ggctagagat cctgatgatt ggtctcgtct
    15121 ggcgctccat ggagcaccca gggaagctac tgtttgctcc taacttgctc ttggacagga
    15181 accagggaaa atgtgtagag ggcatggtgg agatcttcga catgctgctg gctacatcat
    15241 ctcggttccg catgatgaat ctgcagggag aggagtttgt gtgcctcaaa tctattattt
    15301 tgcttaattc tggagtgtac acatttctgt ccagcaccct gaagtctctg gaagagaagg
    15361 accatatcca ccgagtcctg gacaagatca cagacacttt gatccacctg atggccaagg
    15421 caggcctgac cctgcagcag cagcaccagc ggctggccca gctcctcctc atcctctccc
    15481 acatcaggca catgagtaac aaaggcatgg agcatctgta cagcatgaag tgcaagaacg
    15541 tggtgcccct ctatgacctg ctgctggaga tgctggacgc ccaccgccta catgcgccca
    15601 ctagccgtgg aggggcatcc gtggaggaga cggaccaaag ccacttggcc actgcgggct
    15661 ctacttcatc gcattccttg caaaagtatt acatcacggg ggaggcagag ggtttccctg
    15721 ccacagtctg agagctccct ggcggaattc ccagagatgt tagctgaaat catcactaat
    15781 cagataccaa aatattcaaa tggaaatatc aaaaagcttc tgtttcatca aaaatgactc
    15841 gacctaactg agtaagctag cttgttcgag tattatggca ttgggaaaac tgtttttctt
    15901 gtaccatttg ttgtgcttgt aatttactgt gttttttatt cggttttcgc tatcgaactg
    15961 tgaaatggaa atggatggag aagagttaat gaatgatatg gtccttttgt tcattctcaa
    16021 attaatatta tttgtttttt ctcttatttg ttgtgtgttg aatttgaaat tataagagat
    16081 atgcaaacat tttgttttga gtaaaaatgt gtcaaatcgt ggcctctaat gaccgaagtt
    16141 aatatgagga gtaaaacact agatccccaa acaagcttga aactgaaggc gggaaacgac
    16201 aatctgatca tgagcggaga attaagggag tcacgttatg acccccgccg atgacgcggg
    16261 acaagccgtt ttacgtttgg aactgacaga accgcaacgt tgaaggagcc actcagccgc
    16321 gggtttctgg agtttaatga gctaagcaca tacgtcagaa accattattg cgcgttcaaa
    16381 agtcgcctaa ggtcactatc agctagcaaa tatttcttgt caaaaatgct ccactgacgt
    16441 tccataaatt cccctcggta tccaattaga gtctcatatt cactctcaat ccaaataatc
    16501 tgcaccggat ccgctagagg atctcgacct gcaagatccc ggggggcaat gagatatgaa
    16561 aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt
    16621 ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg
    16681 agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta
    16741 tgtttatcgg
//
