LOCUS       sequence-390446-       15927 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cas9 fused with TLS1 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 15927)
  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 15927)
  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 TLS1
FEATURES             Location/Qualifiers
     source          1..15927
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      917..1169
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        1492..1538
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             1572..1637
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1644..1664
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1689..5789
                     /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             1689..5789
                     /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             5702..5710
                     /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             5790..5837
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(6415..6432)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6415..6431)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6424..6446)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    6653..6677
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             7198..7989
                     /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      8235..8823
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8724..8743
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8977..8994
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    9009..9149
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(9075..9094)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9235..9257
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      9493..9687
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(9753..10826)
                     /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(11255..11884)
                     /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    13215..13239
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     promoter        13430..13476
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             13505..13756
                     /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             13772..14005
                     /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             14015..14950
                     /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             14063..14956
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPGKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATV"
     promoter        15514..15693
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        15546..15729
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
ORIGIN
        1 gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcg
       61 gccgcgctcc cgattccgga agtgcttgac attggggaat tcagcgagag cctgacctat
      121 tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc
      181 gctgttctgc agccggtcgc ggaggccatg gatgcgatcg ctgcggccga tcttagccag
      241 acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat
      301 ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc
      361 gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc
      421 gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc
      481 cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc
      541 gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc
      601 gagcggaggc atccggagct tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt
      661 ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg
      721 cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc
      781 gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga
      841 aaccgacgcc ccagcactcg tccgggatct tggaggtgat gtaacatgat cacaagctga
      901 tcccccgaat ttccccgatc gttcaaacat ttggcaataa agtttcttaa gattgaatcc
      961 tgttgccggt cttgcgatga ttatcatata atttctgttg aattacgtta agcatgtaat
     1021 aattaacatg taatgcatga cgttatttat gagatgggtt tttatgatta gagtcccgca
     1081 attatacatt taatacgcga tagaaaacaa aatatagcgc gcaaactagg ataaattatc
     1141 gcgcgcggtg tcatctatgt tactagatcg gggaattgat cccccctcga cagcttgcat
     1201 gccagcttgg gctgcaggtc gaggctaaaa aactaatcgc attatcatcc cctcgacgta
     1261 ctgtacatat aaccactggt tttatataca gcagtactgt acatataacc actggtttta
     1321 tatacagcag tcgacgtact gtacatataa ccactggttt tatatacagc agtactgtac
     1381 atataaccac tggttttata tacagcagtc gaggtaagat tagatatgga tatgtatatg
     1441 gatatgtata tggtggtaat gccatgtaat atgctcgact ctaggatctt cgcaagaccc
     1501 ttcctctata taaggaagtt catttcattt ggagaggaca cgctgaagct agtcgactct
     1561 agcctcgaat ggattacaag gaccacgacg gggattacaa ggaccacgac attgattaca
     1621 aggatgatga tgacaagatg gctccgaaga agaagaggaa ggttggcatc cacggggtgc
     1681 cagctgctga caagaagtac tcgatcggcc tcgatattgg gactaactct gttggctggg
     1741 ccgtgatcac cgacgagtac aaggtgccct caaagaagtt caaggtcctg ggcaacaccg
     1801 atcggcattc catcaagaag aatctcattg gcgctctcct gttcgacagc ggcgagacgg
     1861 ctgaggctac gcggctcaag cgcaccgccc gcaggcggta cacgcgcagg aagaatcgca
     1921 tctgctacct gcaggagatt ttctccaacg agatggcgaa ggttgacgat tctttcttcc
     1981 acaggctgga ggagtcattc ctcgtggagg aggataagaa gcacgagcgg catccaatct
     2041 tcggcaacat tgtcgacgag gttgcctacc acgagaagta ccctacgatc taccatctgc
     2101 ggaagaagct cgtggactcc acagataagg cggacctccg cctgatctac ctcgctctgg
     2161 cccacatgat taagttcagg ggccatttcc tgatcgaggg ggatctcaac ccggacaata
     2221 gcgatgttga caagctgttc atccagctcg tgcagacgta caaccagctc ttcgaggaga
     2281 accccattaa tgcgtcaggc gtcgacgcga aggctatcct gtccgctagg ctctcgaagt
     2341 ctcggcgcct cgagaacctg atcgcccagc tgccgggcga gaagaagaac ggcctgttcg
     2401 ggaatctcat tgcgctcagc ctggggctca cgcccaactt caagtcgaat ttcgatctcg
     2461 ctgaggacgc caagctgcag ctctccaagg acacatacga cgatgacctg gataacctcc
     2521 tggcccagat cggcgatcag tacgcggacc tgttcctcgc tgccaagaat ctgtcggacg
     2581 ccatcctcct gtctgatatt ctcagggtga acaccgagat tacgaaggct ccgctctcag
     2641 cctccatgat caagcgctac gacgagcacc atcaggatct gaccctcctg aaggcgctgg
     2701 tcaggcagca gctccccgag aagtacaagg agatcttctt cgatcagtcg aagaacggct
     2761 acgctgggta cattgacggc ggggcctctc aggaggagtt ctacaagttc atcaagccga
     2821 ttctggagaa gatggacggc acggaggagc tgctggtgaa gctcaatcgc gaggacctcc
     2881 tgaggaagca gcggacattc gataacggca gcatcccaca ccagattcat ctcggggagc
     2941 tgcacgctat cctgaggagg caggaggact tctacccttt cctcaaggat aaccgcgaga
     3001 agatcgagaa gattctgact ttcaggatcc cgtactacgt cggcccactc gctaggggca
     3061 actcccgctt cgcttggatg acccgcaagt cagaggagac gatcacgccg tggaacttcg
     3121 aggaggtggt cgacaagggc gctagcgctc agtcgttcat cgagaggatg acgaatttcg
     3181 acaagaacct gccaaatgag aaggtgctcc ctaagcactc gctcctgtac gagtacttca
     3241 cagtctacaa cgagctgact aaggtgaagt atgtgaccga gggcatgagg aagccggctt
     3301 tcctgtctgg ggagcagaag aaggccatcg tggacctcct gttcaagacc aaccggaagg
     3361 tcacggttaa gcagctcaag gaggactact tcaagaagat tgagtgcttc gattcggtcg
     3421 agatctctgg cgttgaggac cgcttcaacg cctccctggg gacctaccac gatctcctga
     3481 agatcattaa ggataaggac ttcctggaca acgaggagaa tgaggatatc ctcgaggaca
     3541 ttgtgctgac actcactctg ttcgaggacc gggagatgat cgaggagcgc ctgaagactt
     3601 acgcccatct cttcgatgac aaggtcatga agcagctcaa gaggaggagg tacaccggct
     3661 gggggaggct gagcaggaag ctcatcaacg gcattcggga caagcagtcc gggaagacga
     3721 tcctcgactt cctgaagagc gatggcttcg cgaaccgcaa tttcatgcag ctgattcacg
     3781 atgacagcct cacattcaag gaggatatcc agaaggctca ggtgagcggc cagggggact
     3841 cgctgcacga gcatatcgcg aacctcgctg gctcgccagc tatcaagaag gggattctgc
     3901 agaccgtgaa ggttgtggac gagctggtga aggtcatggg caggcacaag cctgagaaca
     3961 tcgtcattga gatggcccgg gagaatcaga ccacgcagaa gggccagaag aactcacgcg
     4021 agaggatgaa gaggatcgag gagggcatta aggagctggg gtcccagatc ctcaaggagc
     4081 acccggtgga gaacacgcag ctgcagaatg agaagctcta cctgtactac ctccagaatg
     4141 gccgcgatat gtatgtggac caggagctgg atattaacag gctcagcgat tacgacgtcg
     4201 atcatatcgt tccacagtca ttcctgaagg atgactccat tgacaacaag gtcctcacca
     4261 ggtcggacaa gaaccggggc aagtctgata atgttccttc agaggaggtc gttaagaaga
     4321 tgaagaacta ctggcgccag ctcctgaatg ccaagctgat cacgcagcgg aagttcgata
     4381 acctcacaaa ggctgagagg ggcgggctct ctgagctgga caaggcgggc ttcatcaaga
     4441 ggcagctggt cgagacacgg cagatcacta agcacgttgc gcagattctc gactcacgga
     4501 tgaacactaa gtacgatgag aatgacaagc tgatccgcga ggtgaaggtc atcaccctga
     4561 agtcaaagct cgtctccgac ttcaggaagg atttccagtt ctacaaggtt cgggagatca
     4621 acaattacca ccatgcccat gacgcgtacc tgaacgcggt ggtcggcaca gctctgatca
     4681 agaagtaccc aaagctcgag agcgagttcg tgtacgggga ctacaaggtt tacgatgtga
     4741 ggaagatgat cgccaagtcg gagcaggaga ttggcaaggc taccgccaag tacttcttct
     4801 actctaacat tatgaatttc ttcaagacag agatcactct ggccaatggc gagatccgga
     4861 agcgccccct catcgagacg aacggcgaga cgggggagat cgtgtgggac aagggcaggg
     4921 atttcgcgac cgtcaggaag gttctctcca tgccacaagt gaatatcgtc aagaagacag
     4981 aggtccagac tggcgggttc tctaaggagt caattctgcc taagcggaac agcgacaagc
     5041 tcatcgcccg caagaaggac tgggatccga agaagtacgg cgggttcgac agccccactg
     5101 tggcctactc ggtcctggtt gtggcgaagg ttgagaaggg caagtccaag aagctcaaga
     5161 gcgtgaagga gctgctgggg atcacgatta tggagcgctc cagcttcgag aagaacccga
     5221 tcgatttcct ggaggcgaag ggctacaagg aggtgaagaa ggacctgatc attaagctcc
     5281 ccaagtactc actcttcgag ctggagaacg gcaggaagcg gatgctggct tccgctggcg
     5341 agctgcagaa ggggaacgag ctggctctgc cgtccaagta tgtgaacttc ctctacctgg
     5401 cctcccacta cgagaagctc aagggcagcc ccgaggacaa cgagcagaag cagctgttcg
     5461 tcgagcagca caagcattac ctcgacgaga tcattgagca gatttccgag ttctccaagc
     5521 gcgtgatcct ggccgacgcg aatctggata aggtcctctc cgcgtacaac aagcaccgcg
     5581 acaagccaat cagggagcag gctgagaata tcattcatct cttcaccctg acgaacctcg
     5641 gcgcccctgc tgctttcaag tacttcgaca caactatcga tcgcaagagg tacacaagca
     5701 ctaaggaggt cctggacgcg accctcatcc accagtcgat taccggcctc tacgagacgc
     5761 gcatcgacct gtctcagctc gggggcgaca agcggccagc ggcgacgaag aaggcggggc
     5821 aggcgaagaa gaagaagtga attatcagag tggcgcagcg gaagcgtggt gggcccataa
     5881 cccacaggtc ccaggatcga aacctggctc tgatatcgag agagctttcg ttcgtatcat
     5941 cggtttcgac aacgttcgtc aagttcaatg catcagtttc attgcccaca caccagaatc
     6001 ctactaagtt tgagtattat ggcattggaa aagctgtttt cttctatcat ttgttctgct
     6061 tgtaatttac tgtgttcttt cagtttttgt tttcggacat caaaatgcaa atggatggat
     6121 aagagttaat aaatgatatg gtccttttgt tcattctcaa attattatta tctgttgttt
     6181 ttactttaat gggttgaatt taagtaagaa aggaactaac agtgtgatat taaggtgcaa
     6241 tgttagacat ataaaacagt ctttcacctc tctttggtta tgtcttgaat tggtttgttt
     6301 cttcacttat ctgtgtaatc aagtttacta tgagtctatg atcaagtaat tatgcaatca
     6361 agttaagtac agtataggct ttttgtgtcg agggggtacc gagtcgagga attcactggc
     6421 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     6481 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     6541 ccaacagttg cgcagcctga atggcgggta ccgagctcga attcaattcg gcgttaattc
     6601 agtacattaa aaacgtccgc aatgtgttat taagttgtct aagcgtcaat ttgtttacac
     6661 cacaatatat cctgccacca gccagccaac agctccccga ccggcagctc ggcacaaaat
     6721 caccactcga tacaggcagc ccatcagtcc gggacggcgt cagcgggaga gccgttgtaa
     6781 ggcggcagac tttgctcatg ttaccgatgc tattcggaag aacggcaact aagctgccgg
     6841 gtttgaaaca cggatgatct cgcggagggt agcatgttga ttgtaacgat gacagagcgt
     6901 tgctgcctgt gatcaattcg ggcacgaacc cagtggacat aagcctgttc ggttcgtaag
     6961 ctgtaatgca agtagcgtat gcgctcacgc aactggtcca gaaccttgac cgaacgcagc
     7021 ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg actgtttttt tggggtacag
     7081 tctatgcctc gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta
     7141 tggagcagca acgatgttac gcagcagggc agtcgcccta aaacaaagtt aaacatcatg
     7201 ggggaagcgg tgatcgccga agtatcgact caactatcag aggtagttgg cgtcatcgag
     7261 cgccatctcg aaccgacgtt gctggccgta catttgtacg gctccgcagt ggatggcggc
     7321 ctgaagccac acagtgatat tgatttgctg gttacggtga ccgtaaggct tgatgaaaca
     7381 acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg cttcccctgg agagagcgag
     7441 attctccgcg ctgtagaagt caccattgtt gtgcacgacg acatcattcc gtggcgttat
     7501 ccagctaagc gcgaactgca atttggagaa tggcagcgca atgacattct tgcaggtatc
     7561 ttcgagccag ccacgatcga cattgatctg gctatcttgc tgacaaaagc aagagaacat
     7621 agcgttgcct tggtaggtcc agcggcggag gaactctttg atccggttcc tgaacaggat
     7681 ctatttgagg cgctaaatga aaccttaacg ctatggaact cgccgcccga ctgggctggc
     7741 gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt acagcgcagt aaccggcaaa
     7801 atcgcgccga aggatgtcgc tgccgactgg gcaatggagc gcctgccggc ccagtatcag
     7861 cccgtcatac ttgaagctag acaggcttat cttggacaag aagaagatcg cttggcctcg
     7921 cgcgcagatc agttggaaga atttgtccac tacgtgaaag gcgagatcac caaggtagtc
     7981 ggcaaataat gtctagctag aaattcgttc aagccgacgc cgcttcgcgg cgcggcttaa
     8041 ctcaagcgtt agatgcacta agcacataat tgctcacagc caaactatca ggtcaagtct
     8101 gcttttatta tttttaagcg tgcataataa gccctacaca aattgggaga tatatcatgc
     8161 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     8221 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     8281 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     8341 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     8401 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     8461 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     8521 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     8581 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     8641 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     8701 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     8761 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     8821 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     8881 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     8941 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     9001 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     9061 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
     9121 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
     9181 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     9241 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ggtgccttga
     9301 tgtgggcgcc ggcggtcgag tggcgacggc gcggcttgtc cgcgccctgg tagattgcct
     9361 ggccgtaggc cagccatttt tgagcggcca gcggccgcga taggccgacg cgaagcggcg
     9421 gggcgtaggg agcgcagcga ccgaagggta ggcgcttttt gcagctcttc ggctgtgcgc
     9481 tggccagaca gttatgcaca ggccaggcgg gttttaagag ttttaataag ttttaaagag
     9541 ttttaggcgg aaaaatcgcc ttttttctct tttatatcag tcacttacat gtgtgaccgg
     9601 ttcccaatgt acggctttgg gttcccaatg tacgggttcc ggttcccaat gtacggcttt
     9661 gggttcccaa tgtacgtgct atccacagga aagagacctt ttcgaccttt ttcccctgct
     9721 agggcaattt gccctagcat ctgctccgta cattaggaac cggcggatgc ttcgccctcg
     9781 atcaggttgc ggtagcgcat gactaggatc gggccagcct gccccgcctc ctccttcaaa
     9841 tcgtactccg gcaggtcatt tgacccgatc agcttgcgca cggtgaaaca gaacttcttg
     9901 aactctccgg cgctgccact gcgttcgtag atcgtcttga acaaccatct ggcttctgcc
     9961 ttgcctgcgg cgcggcgtgc caggcggtag agaaaacggc cgatgccggg atcgatcaaa
    10021 aagtaatcgg ggtgaaccgt cagcacgtcc gggttcttgc cttctgtgat ctcgcggtac
    10081 atccaatcag ctagctcgat ctcgatgtac tccggccgcc cggtttcgct ctttacgatc
    10141 ttgtagcggc taatcaaggc ttcaccctcg gataccgtca ccaggcggcc gttcttggcc
    10201 ttcttcgtac gctgcatggc aacgtgcgtg gtgtttaacc gaatgcaggt ttctaccagg
    10261 tcgtctttct gctttccgcc atcggctcgc cggcagaact tgagtacgtc cgcaacgtgt
    10321 ggacggaaca cgcggccggg cttgtctccc ttcccttccc ggtatcggtt catggattcg
    10381 gttagatggg aaaccgccat cagtaccagg tcgtaatccc acacactggc catgccggcc
    10441 ggccctgcgg aaacctctac gtgcccgtct ggaagctcgt agcggatcac ctcgccagct
    10501 cgtcggtcac gcttcgacag acggaaaacg gccacgtcca tgatgctgcg actatcgcgg
    10561 gtgcccacgt catagagcat cggaacgaaa aaatctggtt gctcgtcgcc cttgggcggc
    10621 ttcctaatcg acggcgcacc ggctgccggc ggttgccggg attctttgcg gattcgatca
    10681 gcggccgctt gccacgattc accggggcgt gcttctgcct cgatgcgttg ccgctgggcg
    10741 gcctgcgcgg ccttcaactt ctccaccagg tcatcaccca gcgccgcgcc gatttgtacc
    10801 gggccggatg gtttgcgacc gctcacgccg attcctcggg cttgggggtt ccagtgccat
    10861 tgcagggccg gcagacaacc cagccgctta cgcctggcca accgcccgtt cctccacaca
    10921 tggggcattc cacggcgtcg gtgcctggtt gttcttgatt ttccatgccg cctcctttag
    10981 ccgctaaaat tcatctactc atttattcat ttgctcattt actctggtag ctgcgcgatg
    11041 tattcagata gcagctcggt aatggtcttg ccttggcgta ccgcgtacat cttcagcttg
    11101 gtgtgatcct ccgccggcaa ctgaaagttg acccgcttca tggctggcgt gtctgccagg
    11161 ctggccaacg ttgcagcctt gctgctgcgt gcgctcggac ggccggcact tagcgtgttt
    11221 gtgcttttgc tcattttctc tttacctcat taactcaaat gagttttgat ttaatttcag
    11281 cggccagcgc ctggacctcg cgggcagcgt cgccctcggg ttctgattca agaacggttg
    11341 tgccggcggc ggcagtgcct gggtagctca cgcgctgcgt gatacgggac tcaagaatgg
    11401 gcagctcgta cccggccagc gcctcggcaa cctcaccgcc gatgcgcgtg cctttgatcg
    11461 cccgcgacac gacaaaggcc gcttgtagcc ttccatccgt gacctcaatg cgctgcttaa
    11521 ccagctccac caggtcggcg gtggcccata tgtcgtaagg gcttggctgc accggaatca
    11581 gcacgaagtc ggctgccttg atcgcggaca cagccaagtc cgccgcctgg ggcgctccgt
    11641 cgatcactac gaagtcgcgc cggccgatgg ccttcacgtc gcggtcaatc gtcgggcggt
    11701 cgatgccgac aacggttagc ggttgatctt cccgcacggc cgcccaatcg cgggcactgc
    11761 cctggggatc ggaatcgact aacagaacat cggccccggc gagttgcagg gcgcgggcta
    11821 gatgggttgc gatggtcgtc ttgcctgacc cgcctttctg gttaagtaca gcgataacct
    11881 tcatgcgttc cccttgcgta tttgtttatt tactcatcgc atcatatacg cagcgaccgc
    11941 atgacgcaag ctgttttact caaatacaca tcaccttttt agacggcggc gctcggtttc
    12001 ttcagcggcc aagctggccg gccaggccgc cagcttggca tcagacaaac cggccaggat
    12061 ttcatgcagc cgcacggttg agacgtgcgc gggcggctcg aacacgtacc cggccgcgat
    12121 catctccgcc tcgatctctt cggtaatgaa aaacggttcg tcctggccgt cctggtgcgg
    12181 tttcatgctt gttcctcttg gcgttcattc tcggcggccg ccagggcgtc ggcctcggtc
    12241 aatgcgtcct cacggaaggc accgcgccgc ctggcctcgg tgggcgtcac ttcctcgctg
    12301 cgctcaagtg cgcggtacag ggtcgagcga tgcacgccaa gcagtgcagc cgcctctttc
    12361 acggtgcggc cttcctggtc gatcagctcg cgggcgtgcg cgatctgtgc cggggtgagg
    12421 gtagggcggg ggccaaactt cacgcctcgg gccttggcgg cctcgcgccc gctccgggtg
    12481 cggtcgatga ttagggaacg ctcgaactcg gcaatgccgg cgaacacggt caacaccatg
    12541 cggccggccg gcgtggtggt gtcggcccac ggctctgcca ggctacgcag gcccgcgccg
    12601 gcctcctgga tgcgctcggc aatgtccagt aggtcgcggg tgctgcgggc caggcggtct
    12661 agcctggtca ctgtcacaac gtcgccaggg cgtaggtggt caagcatcct ggccagctcc
    12721 gggcggtcgc gcctggtgcc ggtgatcttc tcggaaaaca gcttggtgca gccggccgcg
    12781 tgcagttcgg cccgttggtt ggtcaagtcc tggtcgtcgg tgctgacgcg ggcatagccc
    12841 agcaggccag cggcggcgct cttgttcatg gcgtaatgtc tccggttcta gtcgcaagta
    12901 ttctacttta tgcgactaaa acacgcgaca agaaaacgcc aggaaaaggg cagggcggca
    12961 gcctgtcgcg taacttagga cttgtgcgac atgtcgtttt cagaagacgg ctgcactgaa
    13021 cgtcagaagc cgactgcact atagcagcgg aggggttgga tcaaagtact ttaaagtact
    13081 ttaaagtact ttaaagtact ttgatcccga ggggaaccct gtggttggca tgcacataca
    13141 aatggacgaa cggataaacc ttttcacgcc cttttaaata tccgattatt ctaataaacg
    13201 ctcttttctc ttaggtttac ccgccaatat atcctgtcaa acactgatag tttaaactga
    13261 aggcgggaaa cgacaatctg atccaagctc aagctaagct tgcatgcctg caggatatcg
    13321 tggatccaag cttgccacgt gccgccacgt gccgccacgt gccgccacgt gcctctagag
    13381 gatccatctc cactgacgta agggatgacg cacaatccca ctatccttcg caagaccctt
    13441 cctctatata aggaagttca tttcatttgg agaggacacg ctgggatccc caattccggg
    13501 cggaatgaaa gcgttaacgg ccaggcaaca agaggtgttt gatctcatcc gtgatcacat
    13561 cagccagaca ggtatgccgc cgacgcgtgc ggaaatcgcg cagcgtttgg ggttccgttc
    13621 cccaaacgcg gctgaagaac atctgaaggc gctggcacgc aaaggcgtta ttgaaattgt
    13681 ttccggcgca tcacgcggga ttcgtctgtt gcaggaagag gaagaagggt tgccgctggt
    13741 aggtcgtgtg gctgccggtg aaccgtcgag cgcccccccg accgatgtca gcctggggga
    13801 cgagctccac ttagacggcg aggacgtggc gatggcgcat gccgacgcgc tagacgattt
    13861 cgatctggac atgttggggg acggggattc cccgggtccg ggatttaccc cccacgactc
    13921 cgccccctac ggcgctctgg atatggccga cttcgagttt gagcagatgt ttaccgatgc
    13981 ccttggaatt gacgagtacg gtggggatcc gtctgctgga gacatgagag ctgccaacct
    14041 ttggccaagc ccgctcatga tcaaacgctc taagaagaac agcctggcct tgtccctgac
    14101 ggccgaccag atggtcagtg ccttgttgga tgctgagccc cccatactct attccgagta
    14161 tgatcctacc agacccttca gtgaagcttc gatgatgggc ttactgacca acctggcaga
    14221 cagggagctg gttcacatga tcaactgggc gaagagggtg ccaggctttg tggatttgac
    14281 cctccatgat caggtccacc ttctagaatg tgcctggcta gagatcctga tgattggtct
    14341 cgtctggcgc tccatggagc acccagggaa gctactgttt gctcctaact tgctcttgga
    14401 caggaaccag ggaaaatgtg tagagggcat ggtggagatc ttcgacatgc tgctggctac
    14461 atcatctcgg ttccgcatga tgaatctgca gggagaggag tttgtgtgcc tcaaatctat
    14521 tattttgctt aattctggag tgtacacatt tctgtccagc accctgaagt ctctggaaga
    14581 gaaggaccat atccaccgag tcctggacaa gatcacagac actttgatcc acctgatggc
    14641 caaggcaggc ctgaccctgc agcagcagca ccagcggctg gcccagctcc tcctcatcct
    14701 ctcccacatc aggcacatga gtaacaaagg catggagcat ctgtacagca tgaagtgcaa
    14761 gaacgtggtg cccctctatg acctgctgct ggagatgctg gacgcccacc gcctacatgc
    14821 gcccactagc cgtggagggg catccgtgga ggagacggac caaagccact tggccactgc
    14881 gggctctact tcatcgcatt ccttgcaaaa gtattacatc acgggggagg cagagggttt
    14941 ccctgccaca gtctgagagc tccctggcgg aattcccaga gatgttagct gaaatcatca
    15001 ctaatcagat accaaaatat tcaaatggaa atatcaaaaa gcttctgttt catcaaaaat
    15061 gactcgacct aactgagtaa gctagcttgt tcgagtatta tggcattggg aaaactgttt
    15121 ttcttgtacc atttgttgtg cttgtaattt actgtgtttt ttattcggtt ttcgctatcg
    15181 aactgtgaaa tggaaatgga tggagaagag ttaatgaatg atatggtcct tttgttcatt
    15241 ctcaaattaa tattatttgt tttttctctt atttgttgtg tgttgaattt gaaattataa
    15301 gagatatgca aacattttgt tttgagtaaa aatgtgtcaa atcgtggcct ctaatgaccg
    15361 aagttaatat gaggagtaaa acactagatc cccaaacaag cttgaaactg aaggcgggaa
    15421 acgacaatct gatcatgagc ggagaattaa gggagtcacg ttatgacccc cgccgatgac
    15481 gcgggacaag ccgttttacg tttggaactg acagaaccgc aacgttgaag gagccactca
    15541 gccgcgggtt tctggagttt aatgagctaa gcacatacgt cagaaaccat tattgcgcgt
    15601 tcaaaagtcg cctaaggtca ctatcagcta gcaaatattt cttgtcaaaa atgctccact
    15661 gacgttccat aaattcccct cggtatccaa ttagagtctc atattcactc tcaatccaaa
    15721 taatctgcac cggatccgct agaggatctc gacctgcaag atcccggggg gcaatgagat
    15781 atgaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac
    15841 agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat
    15901 gtaggagggc gtggatatgt cctgcgg
//
