LOCUS       sequence-173269-       17990 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR/Cas9 in Arabidopsis with seed a fluorescent reporter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17990)
  AUTHORS   Tsutsui H, Higashiyama T
  TITLE     pKAMA-ITACHI vectors for highly efficient CRISPR/Cas9-mediated gene 
            knockout in Arabidopsis thaliana.
  JOURNAL   Plant Cell Physiol. 2016 Nov 17. pii: pcw191.
  PUBMED    27856772
REFERENCE   2  (bases 1 to 17990)
  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: "Plant Cell 
            Physiol. 2016 Nov 17. pii: pcw191."
COMMENT     Sequence Label: pKIR1.0
FEATURES             Location/Qualifiers
     source          1..17990
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      19..28
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    41..65
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1745..1810
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1817..1837
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1862..5962
                     /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             1862..5962
                     /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             5963..6010
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      6024..6272
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     protein_bind    complement(6281..6305)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             8451..9146
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSDMALK
                     LVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAVARYC
                     DLPSKLGHK"
     terminator      9147..9398
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    9502..9526
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             10857..11486
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             11915..12988
                     /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"
     rep_origin      13054..13248
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(13484..13506)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    13592..13732
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     13647..13666
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(13747..13764)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13918..14506)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13998..14017)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(14752..15543)
                     /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"
     misc_feature    16064..16088
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(16181..16433)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(16631..17656)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(17692..17871)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
ORIGIN
        1 cgacgtcgca tgctcccggc cgccatggcc gcgggatatc acaagtttgt acaaaaaagc
       61 aggctccgcg gccgcgatcc ctcaactttt gattcgctat ttgcagtgca cctgtggcgt
      121 tcatcacatc ttttgtgaca ctgtttgcac tggtcattgc tattacaaag gaccttcctg
      181 atgttgaagg agatcgaaag taagtaactg cacgcataac cattttcttt ccgctctttg
      241 gctcaatcca tttgacagtc aaagacaatg tttaaccagc tccgtttgat atattgtctt
      301 tatgtgtttg ttcaagcatg tttagttaat catgcctttg attgatcttg aataggttcc
      361 aaatatcaac cctggcaaca aaacttggag tgagaaacat tgcattcctc ggttctggac
      421 ttctgctagt aaattatgtt tcagccatat cactagcttt ctacatgcct caggtgaatt
      481 catctatttc cgtcttaact atttcggtta attaaagcac gaacaccatt actgcatgta
      541 gaagcttgat aaactatcgc caccaattta tttttgttgc gatattgtta ctttcctcag
      601 tatgcagctt tgaaaagacc aaccctctta tcctttaaca atgaacaggt ttttagaggt
      661 agcttgatga ttcctgcaca tgtgatcttg gcttcaggct taattttcca ggtaaagcat
      721 tatgagatac tcttatatct cttacatact tttgagataa tgcacaagaa cttcataact
      781 atatgcttta gtttctgcat ttgacactgc caaattcatt aatctctaat atctttgttg
      841 ttgatctttg gtagacatgg gtactagaaa aagcaaacta caccaaggta aaatactttt
      901 gtacaaacat aaactcgtta tcacggaaca tcaatggagt gtatatctaa cggagtgtag
      961 aaacatttga ttattgcagg aagctatctc aggatattat cggtttatat ggaatctctt
     1021 ctacgcagag tatctgttat tccccttcct ctagctttca atttcatggt gaggatatgc
     1081 agttttcttt gtatatcatt cttcttcttc tttgtagctt ggagtcaaaa tcggttcctt
     1141 catgtacata catcaaggat atgtccttct gaatttttat atcttgcaat aaaaatgctt
     1201 gtaccaattg aaacaccagc tttttgagtt ctatgatcac tgacttggtt ctaaccaaaa
     1261 aaaaaaaatg tttaatttac atatctaaaa gtaggtttag ggaaacctaa acagtaaaat
     1321 atttgtatat tattcgaatt tcactcatca taaaaactta aattgcacca taaaattttg
     1381 ttttactatt aatgatgtaa tttgtgtaac ttaagataaa aataatattc cgtaagttaa
     1441 ccggctaaaa ccacgtataa accagggaac ctgttaaacc ggttctttac tggataaaga
     1501 aatgaaagcc catgtagaca gctccattag agcccaaacc ctaaatttct catctatata
     1561 aaaggagtga cattagggtt tttgttcgtc ctcttaaagc ttctcgtttt ctctgccgtc
     1621 tctctcattc gcgcgacgca aacgatcttc aggtgatctt ctttctccaa atcctctctc
     1681 ataactctga tttcgtactt gtgtatttga gctcacgctc tgtttctctc accacagccc
     1741 catggactat aaggaccacg acggagacta caaggatcat gatattgatt acaaagacga
     1801 tgacgataag atggccccaa agaagaagcg gaaggtcggt atccacggag tcccagcagc
     1861 cgacaagaag tacagcatcg gcctggacat cggcaccaac tctgtgggct gggccgtgat
     1921 caccgacgag tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca
     1981 cagcatcaag aagaacctga tcggagccct gctgttcgac agcggcgaaa cagccgaggc
     2041 cacccggctg aagagaaccg ccagaagaag atacaccaga cggaagaacc ggatctgcta
     2101 tctgcaagag atcttcagca acgagatggc caaggtggac gacagcttct tccacagact
     2161 ggaagagtcc ttcctggtgg aagaggataa gaagcacgag cggcacccca tcttcggcaa
     2221 catcgtggac gaggtggcct accacgagaa gtaccccacc atctaccacc tgagaaagaa
     2281 actggtggac agcaccgaca aggccgacct gcggctgatc tatctggccc tggcccacat
     2341 gatcaagttc cggggccact tcctgatcga gggcgacctg aaccccgaca acagcgacgt
     2401 ggacaagctg ttcatccagc tggtgcagac ctacaaccag ctgttcgagg aaaaccccat
     2461 caacgccagc ggcgtggacg ccaaggccat cctgtctgcc agactgagca agagcagacg
     2521 gctggaaaat ctgatcgccc agctgcccgg cgagaagaag aatggcctgt tcggaaacct
     2581 gattgccctg agcctgggcc tgacccccaa cttcaagagc aacttcgacc tggccgagga
     2641 tgccaaactg cagctgagca aggacaccta cgacgacgac ctggacaacc tgctggccca
     2701 gatcggcgac cagtacgccg acctgtttct ggccgccaag aacctgtccg acgccatcct
     2761 gctgagcgac atcctgagag tgaacaccga gatcaccaag gcccccctga gcgcctctat
     2821 gatcaagaga tacgacgagc accaccagga cctgaccctg ctgaaagctc tcgtgcggca
     2881 gcagctgcct gagaagtaca aagagatttt cttcgaccag agcaagaacg gctacgccgg
     2941 ctacattgac ggcggagcca gccaggaaga gttctacaag ttcatcaagc ccatcctgga
     3001 aaagatggac ggcaccgagg aactgctcgt gaagctgaac agagaggacc tgctgcggaa
     3061 gcagcggacc ttcgacaacg gcagcatccc ccaccagatc cacctgggag agctgcacgc
     3121 cattctgcgg cggcaggaag atttttaccc attcctgaag gacaaccggg aaaagatcga
     3181 gaagatcctg accttccgca tcccctacta cgtgggccct ctggccaggg gaaacagcag
     3241 attcgcctgg atgaccagaa agagcgagga aaccatcacc ccctggaact tcgaggaagt
     3301 ggtggacaag ggcgcttccg cccagagctt catcgagcgg atgaccaact tcgataagaa
     3361 cctgcccaac gagaaggtgc tgcccaagca cagcctgctg tacgagtact tcaccgtgta
     3421 taacgagctg accaaagtga aatacgtgac cgagggaatg agaaagcccg ccttcctgag
     3481 cggcgagcag aaaaaggcca tcgtggacct gctgttcaag accaaccgga aagtgaccgt
     3541 gaagcagctg aaagaggact acttcaagaa aatcgagtgc ttcgactccg tggaaatctc
     3601 cggcgtggaa gatcggttca acgcctccct gggcacatac cacgatctgc tgaaaattat
     3661 caaggacaag gacttcctgg acaatgagga aaacgaggac attctggaag atatcgtgct
     3721 gaccctgaca ctgtttgagg acagagagat gatcgaggaa cggctgaaaa cctatgccca
     3781 cctgttcgac gacaaagtga tgaagcagct gaagcggcgg agatacaccg gctggggcag
     3841 gctgagccgg aagctgatca acggcatccg ggacaagcag tccggcaaga caatcctgga
     3901 tttcctgaag tccgacggct tcgccaacag aaacttcatg cagctgatcc acgacgacag
     3961 cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg atagcctgca
     4021 cgagcacatt gccaatctgg ccggcagccc cgccattaag aagggcatcc tgcagacagt
     4081 gaaggtggtg gacgagctcg tgaaagtgat gggccggcac aagcccgaga acatcgtgat
     4141 cgaaatggcc agagagaacc agaccaccca gaagggacag aagaacagcc gcgagagaat
     4201 gaagcggatc gaagagggca tcaaagagct gggcagccag atcctgaaag aacaccccgt
     4261 ggaaaacacc cagctgcaga acgagaagct gtacctgtac tacctgcaga atgggcggga
     4321 tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc gactacgatg tggaccatat
     4381 cgtgcctcag agctttctga aggacgactc catcgacaac aaggtgctga ccagaagcga
     4441 caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa
     4501 ctactggcgg cagctgctga acgccaagct gattacccag agaaagttcg acaatctgac
     4561 caaggccgag agaggcggcc tgagcgaact ggataaggcc ggcttcatca agagacagct
     4621 ggtggaaacc cggcagatca caaagcacgt ggcacagatc ctggactccc ggatgaacac
     4681 taagtacgac gagaatgaca agctgatccg ggaagtgaaa gtgatcaccc tgaagtccaa
     4741 gctggtgtcc gatttccgga aggatttcca gttttacaaa gtgcgcgaga tcaacaacta
     4801 ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga accgccctga tcaaaaagta
     4861 ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat
     4921 gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc aagtacttct tctacagcaa
     4981 catcatgaac tttttcaaga ccgagattac cctggccaac ggcgagatcc ggaagcggcc
     5041 tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg gataagggcc gggattttgc
     5101 caccgtgcgg aaagtgctga gcatgcccca agtgaatatc gtgaaaaaga ccgaggtgca
     5161 gacaggcggc ttcagcaaag agtctatcct gcccaagagg aacagcgata agctgatcgc
     5221 cagaaagaag gactgggacc ctaagaagta cggcggcttc gacagcccca ccgtggccta
     5281 ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc aagaaactga agagtgtgaa
     5341 agagctgctg gggatcacca tcatggaaag aagcagcttc gagaagaatc ccatcgactt
     5401 tctggaagcc aagggctaca aagaagtgaa aaaggacctg atcatcaagc tgcctaagta
     5461 ctccctgttc gagctggaaa acggccggaa gagaatgctg gcctctgccg gcgaactgca
     5521 gaagggaaac gaactggccc tgccctccaa atatgtgaac ttcctgtacc tggccagcca
     5581 ctatgagaag ctgaagggct cccccgagga taatgagcag aaacagctgt ttgtggaaca
     5641 gcacaagcac tacctggacg agatcatcga gcagatcagc gagttctcca agagagtgat
     5701 cctggccgac gctaatctgg acaaagtgct gtccgcctac aacaagcacc gggataagcc
     5761 catcagagag caggccgaga atatcatcca cctgtttacc ctgaccaatc tgggagcccc
     5821 tgccgccttc aagtactttg acaccaccat cgaccggaag aggtacacca gcaccaaaga
     5881 ggtgctggac gccaccctga tccaccagag catcaccggc ctgtacgaga cacggatcga
     5941 cctgtctcag ctgggaggcg acaaaaggcc ggcggccacg aaaaaggccg gccaggcaaa
     6001 aaagaaaaag taaaagggtg ggatatgaag atgaagatga aatatttggt gtgtcaaata
     6061 aaaagcttgt gtgcttaagt ttgtgttttt ttcttggctt gttgtgttat gaatttgtgg
     6121 ctttttctaa tattaaatga atgtaagatc tcattataat gaataaacaa atgtttctat
     6181 aatccattgt gaatgttttg ttggatctct tctgcagcat ataactactg tatgtgctat
     6241 ggtatggact atggaatatg attaaagata aggcgcgccg acccagcttt cttgtacaaa
     6301 gtggtgatat cactagtgta tgtaggtata gtaacatgaa caagaacaga ctcaagtaca
     6361 agatcgcata cgaaaatgga aatggcaatg tcacttccac ataatcaaac acgaatcctc
     6421 atatcaacaa ggcctgagat tctaactagc tcataacaac ttagccaata gttacttgag
     6481 actaccaaat gtatgtagaa ctaaagacta agggacagag agttcgtcta aacaggtgaa
     6541 tctagtcgtt gttatctaat aaacaattca gccccaaatg cagaacacac atagagctct
     6601 ctattgattc aaattacgat ctgatactga taacgtctag atttttaggg ttaaagcaat
     6661 caatcacctg acgattcaag gtggttggat catgacgatt ccagaaaaca tcaagcaagc
     6721 tctcaaagct acactctttg ggatcatact gaactctaac aacctcgtta tgtcccgtag
     6781 tgccagtaca gacatcctcg taactcggat tgtgcacgat gccatggcta tacccaacct
     6841 cggtcttggt cacaccagga actctctggt aagctagctc cactccccag aaacaaccgg
     6901 cgccaaattg cgcgaattgc tgacctgaag acggaacatc atcgtcgggt ccttgggcga
     6961 ttgcggcgga agatgggtca gcttgggctt gaggacgaga cccgaatccg agtctgttga
     7021 aaaggttgtt cattggggat ttgtatacgg agattggtcg tcgagaggtt tgagggaaag
     7081 gacaaatggg tttggctctg gagaaagaga gtgcggcttt agagagagaa ttgagaggtt
     7141 tagagagaga tgcggcggcg atgagcggag gagagacgac gaggacctgc attatcaaag
     7201 cagtgacgtg gtgaaatttg gaacttttaa gaggcagata gatttattat ttgtatccat
     7261 tttcttcatt gttctagaat gtcgcggaac aaattttaaa actaaatcct aaatttttct
     7321 aattttgttg ccaatagtgg atatgtgggc cgtatagaag gaatctattg aaggcccaaa
     7381 cccatactga cgagcccaaa ggttcgtttt gcgttttatg tttcggttcg atgccaacgc
     7441 cacattctga gctaggcaaa aaacaaacgt gtctttgaat agactcctct cgttaacaca
     7501 tgcagcggct gcatggtgac gccattaaca cgtggcctac aattgcatga tgtctccatt
     7561 gacacgtgac ttctcgtctc ctttcttaat atatctaaca aacactccta cctcttccaa
     7621 aatatataca catctttttg atcaatctct cattcaaaat ctcattctct ctagtaaaca
     7681 agaacaaaaa aatggcggat acagctagag gaacccatca cgatatcatc ggcagagacc
     7741 agtacccgat gatgggccga gaccgagacc agtaccagat gtccggacga ggatctgact
     7801 actccaagtc taggcagatt gctaaagctg caactgctgt cacagctggt ggttccctcc
     7861 ttgttctctc cagccttacc cttgttggaa ctgtcatagc tttgactgtt gcaacacctc
     7921 tgctcgttat cttcagccca atccttgtcc cggctctcat cacagttgca ctcctcatca
     7981 ccggttttct ttcctctgga gggtttggca ttgccgctat aaccgttttc tcttggattt
     8041 acaagtaagc acacatttat catcttactt cataattttg tgcaatatgt gcatgcatgt
     8101 gttgagccag tagctttgga tcaatttttt tggtcgaata acaaatgtaa caataagaaa
     8161 ttgcaaattc tagggaacat ttggttaact aaatacgaaa tttgacctag ctagcttgaa
     8221 tgtgtctgtg tatatcatct atataggtaa aatgcttggt atgataccta ttgattgtga
     8281 ataggtacgc aacgggagag cacccacagg gatcagacaa gttggacagt gcaaggatga
     8341 agttgggaag caaagctcag gatctgaaag acagagctca gtactacgga cagcaacata
     8401 ctggtgggga acatgaccgt gaccgtactc gtggtggcca gcacactact atgagcgagc
     8461 tgattaagga gaacatgcac atgaagctgt acatggaggg caccgtgaac aaccaccact
     8521 tcaagtgcac atccgagggc gaaggcaagc cctacgaggg cacccagacc atgagaatca
     8581 aggtggtcga gggcggccct ctccccttcg ccttcgacat cctggctacc agcttcatgt
     8641 acggcagcag aaccttcatc aaccacaccc agggcatccc cgacttcttt aagcagtcct
     8701 tccctgaggg cttcacatgg gagagagtca ccacatacga agacgggggc gtgctgaccg
     8761 ctacccagga caccagcctc caggacggct gcctcatcta caacgtcaag atcagagggg
     8821 tgaacttccc atccaacggc cctgtgatgc agaagaaaac actcggctgg gaggccaaca
     8881 ccgagatgct gtaccccgct gacggcggcc tggaaggcag aagcgacatg gccctgaagc
     8941 tcgtgggcgg gggccacctg atctgcaact tcaagaccac atacagatcc aagaaacccg
     9001 ctaagaacct caagatgccc ggcgtctact atgtggacca cagactggaa agaatcaagg
     9061 aggccgacaa agagacctac gtcgagcagc acgaggtggc tgtggccaga tactgcgacc
     9121 tccctagcaa actggggcac aagtgaatcg ttcaaacatt tggcaataaa gtttcttaag
     9181 attgaatcct gttgccggtc ttgcgatgat tatcatataa tttctgttga attacgttaa
     9241 gcatgtaata attaacatgt aatgcatgac gttatttatg agatgggttt ttatgattag
     9301 agtcccgcaa ttatacattt aatacgcgat agaaaacaaa atatagcgcg caaactagga
     9361 taaattatcg cgcgcggtgt catctatgtt actagatcac tagtgcggcc gcctgcaggt
     9421 cgaccatatg ggagagctca agcttagctt gagcttggat cagattgtcg tttcccgcct
     9481 tcagtttaaa ctatcagtgt ttgacaggat atattggcgg gtaaacctaa gagaaaagag
     9541 cgtttattag aataatcgga tatttaaaag ggcgtgaaaa ggtttatccg ttcgtccatt
     9601 tgtatgtgca tgccaaccac agggttcccc tcgggatcaa agtactttaa agtactttaa
     9661 agtactttaa agtactttga tccaacccct ccgctgctat agtgcagtcg gcttctgacg
     9721 ttcagtgcag ccgtcttctg aaaacgacat gtcgcacaag tcctaagtta cgcgacaggc
     9781 tgccgccctg cccttttcct ggcgttttct tgtcgcgtgt tttagtcgca taaagtagaa
     9841 tacttgcgac tagaaccgga gacattacgc catgaacaag agcgccgccg ctggcctgct
     9901 gggctatgcc cgcgtcagca ccgacgacca ggacttgacc aaccaacggg ccgaactgca
     9961 cgcggccggc tgcaccaagc tgttttccga gaagatcacc ggcaccaggc gcgaccgccc
    10021 ggagctggcc aggatgcttg accacctacg ccctggcgac gttgtgacag tgaccaggct
    10081 agaccgcctg gcccgcagca cccgcgacct actggacatt gccgagcgca tccaggaggc
    10141 cggcgcgggc ctgcgtagcc tggcagagcc gtgggccgac accaccacgc cggccggccg
    10201 catggtgttg accgtgttcg ccggcattgc cgagttcgag cgttccctaa tcatcgaccg
    10261 cacccggagc gggcgcgagg ccgccaaggc ccgaggcgtg aagtttggcc cccgccctac
    10321 cctcaccccg gcacagatcg cgcacgcccg cgagctgatc gaccaggaag gccgcaccgt
    10381 gaaagaggcg gctgcactgc ttggcgtgca tcgctcgacc ctgtaccgcg cacttgagcg
    10441 cagcgaggaa gtgacgccca ccgaggccag gcggcgcggt gccttccgtg aggacgcatt
    10501 gaccgaggcc gacgccctgg cggccgccga gaatgaacgc caagaggaac aagcatgaaa
    10561 ccgcaccagg acggccagga cgaaccgttt ttcattaccg aagagatcga ggcggagatg
    10621 atcgcggccg ggtacgtgtt cgagccgccc gcgcacgtct caaccgtgcg gctgcatgaa
    10681 atcctggccg gtttgtctga tgccaagctg gcggcctggc cggccagctt ggccgctgaa
    10741 gaaaccgagc gccgccgtct aaaaaggtga tgtgtatttg agtaaaacag cttgcgtcat
    10801 gcggtcgctg cgtatatgat gcgatgagta aataaacaaa tacgcaaggg gaacgcatga
    10861 aggttatcgc tgtacttaac cagaaaggcg ggtcaggcaa gacgaccatc gcaacccatc
    10921 tagcccgcgc cctgcaactc gccggggccg atgttctgtt agtcgattcc gatccccagg
    10981 gcagtgcccg cgattgggcg gccgtgcggg aagatcaacc gctaaccgtt gtcggcatcg
    11041 accgcccgac gattgaccgc gacgtgaagg ccatcggccg gcgcgacttc gtagtgatcg
    11101 acggagcgcc ccaggcggcg gacttggctg tgtccgcgat caaggcagcc gacttcgtgc
    11161 tgattccggt gcagccaagc ccttacgaca tatgggccac cgccgacctg gtggagctgg
    11221 ttaagcagcg cattgaggtc acggatggaa ggctacaagc ggcctttgtc gtgtcgcggg
    11281 cgatcaaagg cacgcgcatc ggcggtgagg ttgccgaggc gctggccggg tacgagctgc
    11341 ccattcttga gtcccgtatc acgcagcgcg tgagctaccc aggcactgcc gccgccggca
    11401 caaccgttct tgaatcagaa cccgagggcg acgctgcccg cgaggtccag gcgctggccg
    11461 ctgaaattaa atcaaaactc atttgagtta atgaggtaaa gagaaaatga gcaaaagcac
    11521 aaacacgcta agtgccggcc gtccgagcgc acgcagcagc aaggctgcaa cgttggccag
    11581 cctggcagac acgccagcca tgaagcgggt caactttcag ttgccggcgg aggatcacac
    11641 caagctgaag atgtacgcgg tacgccaagg caagaccatt accgagctgc tatctgaata
    11701 catcgcgcag ctaccagagt aaatgagcaa atgaataaat gagtagatga attttagcgg
    11761 ctaaaggagg cggcatggaa aatcaagaac aaccaggcac cgacgccgtg gaatgcccca
    11821 tgtgtggagg aacgggcggt tggccaggcg taagcggctg ggttgtctgc cggccctgca
    11881 atggcactgg aacccccaag cccgaggaat cggcgtgagc ggtcgcaaac catccggccc
    11941 ggtacaaatc ggcgcggcgc tgggtgatga cctggtggag aagttgaagg ccgcgcaggc
    12001 cgcccagcgg caacgcatcg aggcagaagc acgccccggt gaatcgtggc aagcggccgc
    12061 tgatcgaatc cgcaaagaat cccggcaacc gccggcagcc ggtgcgccgt cgattaggaa
    12121 gccgcccaag ggcgacgagc aaccagattt tttcgttccg atgctctatg acgtgggcac
    12181 ccgcgatagt cgcagcatca tggacgtggc cgttttccgt ctgtcgaagc gtgaccgacg
    12241 agctggcgag gtgatccgct acgagcttcc agacgggcac gtagaggttt ccgcagggcc
    12301 ggccggcatg gccagtgtgt gggattacga cctggtactg atggcggttt cccatctaac
    12361 cgaatccatg aaccgatacc gggaagggaa gggagacaag cccggccgcg tgttccgtcc
    12421 acacgttgcg gacgtactca agttctgccg gcgagccgat ggcggaaagc agaaagacga
    12481 cctggtagaa acctgcattc ggttaaacac cacgcacgtt gccatgcagc gtacgaagaa
    12541 ggccaagaac ggccgcctgg tgacggtatc cgagggtgaa gccttgatta gccgctacaa
    12601 gatcgtaaag agcgaaaccg ggcggccgga gtacatcgag atcgagctag ctgattggat
    12661 gtaccgcgag atcacagaag gcaagaaccc ggacgtgctg acggttcacc ccgattactt
    12721 tttgatcgat cccggcatcg gccgttttct ctaccgcctg gcacgccgcg ccgcaggcaa
    12781 ggcagaagcc agatggttgt tcaagacgat ctacgaacgc agtggcagcg ccggagagtt
    12841 caagaagttc tgtttcaccg tgcgcaagct gatcgggtca aatgacctgc cggagtacga
    12901 tttgaaggag gaggcggggc aggctggccc gatcctagtc atgcgctacc gcaacctgat
    12961 cgagggcgaa gcatccgccg gttcctaatg tacggagcag atgctagggc aaattgccct
    13021 agcaggggaa aaaggtcgaa aaggtctctt tcctgtggat agcacgtaca ttgggaaccc
    13081 aaagccgtac attgggaacc ggaacccgta cattgggaac ccaaagccgt acattgggaa
    13141 ccggtcacac atgtaagtga ctgatataaa agagaaaaaa ggcgattttt ccgcctaaaa
    13201 ctctttaaaa cttattaaaa ctcttaaaac ccgcctggcc tgtgcataac tgtctggcca
    13261 gcgcacagcc gaagagctgc aaaaagcgcc tacccttcgg tcgctgcgct ccctacgccc
    13321 cgccgcttcg cgtcggccta tcgcggccgc tggccgctca aaaatggctg gcctacggcc
    13381 aggcaatcta ccagggcgcg gacaagccgc gccgtcgcca ctcgaccgcc ggcgcccaca
    13441 tcaaggcacc ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
    13501 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
    13561 gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata
    13621 gcggagtgta tactggctta actatgcggc atcagagcag attgtactga gagtgcacca
    13681 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc
    13741 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
    13801 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
    13861 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
    13921 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
    13981 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
    14041 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
    14101 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
    14161 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
    14221 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
    14281 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
    14341 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
    14401 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
    14461 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
    14521 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
    14581 gcatgatata tctcccaatt tgtgtagggc ttattatgca cgcttaaaaa taataaaagc
    14641 agacttgacc tgatagtttg gctgtgagca attatgtgct tagtgcatct aacgcttgag
    14701 ttaagccgcg ccgcgaagcg gcgtcggctt gaacgaattt ctagctagac attatttgcc
    14761 gactaccttg gtgatctcgc ctttcacgta gtggacaaat tcttccaact gatctgcgcg
    14821 cgaggccaag cgatcttctt cttgtccaag ataagcctgt ctagcttcaa gtatgacggg
    14881 ctgatactgg gccggcaggc gctccattgc ccagtcggca gcgacatcct tcggcgcgat
    14941 tttgccggtt actgcgctgt accaaatgcg ggacaacgta agcactacat ttcgctcatc
    15001 gccagcccag tcgggcggcg agttccatag cgttaaggtt tcatttagcg cctcaaatag
    15061 atcctgttca ggaaccggat caaagagttc ctccgccgct ggacctacca aggcaacgct
    15121 atgttctctt gcttttgtca gcaagatagc cagatcaatg tcgatcgtgg ctggctcgaa
    15181 gatacctgca agaatgtcat tgcgctgcca ttctccaaat tgcagttcgc gcttagctgg
    15241 ataacgccac ggaatgatgt cgtcgtgcac aacaatggtg acttctacag cgcggagaat
    15301 ctcgctctct ccaggggaag ccgaagtttc caaaaggtcg ttgatcaaag ctcgccgcgt
    15361 tgtttcatca agccttacgg tcaccgtaac cagcaaatca atatcactgt gtggcttcag
    15421 gccgccatcc actgcggagc cgtacaaatg tacggccagc aacgtcggtt cgagatggcg
    15481 ctcgatgacg ccaactacct ctgatagttg agtcgatact tcggcgatca ccgcttcccc
    15541 catgatgttt aactttgttt tagggcgact gccctgctgc gtaacatcgt tgctgctcca
    15601 taacatcaaa catcgaccca cggcgtaacg cgcttgctgc ttggatgccc gaggcataga
    15661 ctgtacccca aaaaaacagt cataacaagc catgaaaacc gccactgcgc cgttaccacc
    15721 gctgcgttcg gtcaaggttc tggaccagtt gcgtgagcgc atacgctact tgcattacag
    15781 cttacgaacc gaacaggctt atgtccactg ggttcgtgcc cgaattgatc acaggcagca
    15841 acgctctgtc atcgttacaa tcaacatgct accctccgcg agatcatccg tgtttcaaac
    15901 ccggcagctt agttgccgtt cttccgaata gcatcggtaa catgagcaaa gtctgccgcc
    15961 ttacaacggc tctcccgctg acgccgtccc ggactgatgg gctgcctgta tcgagtggtg
    16021 attttgtgcc gagctgccgg tcggggagct gttggctggc tggtggcagg atatattgtg
    16081 gtgtaaacaa attgacgctt agacaactta ataacacatt gcggacgttt ttaatgtact
    16141 gaattaacgc cgaattgaat tatcagcttg catgccggtc gatctagtaa catagatgac
    16201 accgcgcgcg ataatttatc ctagtttgcg cgctatattt tgttttctat cgcgtattaa
    16261 atgtataatt gcgggactct aatcataaaa acccatctca taaataacgt catgcattac
    16321 atgttaatta ttacatgctt aacgtaattc aacagaaatt atatgataat catcgcaaga
    16381 ccggcaacag gattcaatct taagaaactt tattgccaaa tgtttgaacg atctgcttga
    16441 ctctagggaa ttaattcctg aatcactgcg accggccctc ccgcgaccca gccgagcgag
    16501 cttagcgaac tgtggacgag aactgtgcca ccaagcgtaa ggccgttctc tcgcatttgc
    16561 cttgctaggc tcgcgcgagt tgctggctga ggcgttctcg aaatcagctc ttgttcggtc
    16621 ggcatctact ctattccttt gccctcggac gagtgctggg gcgtcggttt ccactatcgg
    16681 cgagtacttc tacacagcca tcggtccaga cggccgcgct tctgcgggcg atttgtgtac
    16741 gcccgacagt cccggctccg gatcggacga ttgcgtcgca tcgaccctgc gcccaagctg
    16801 catcatcgaa attgccgtca accaagctct gatagagttg gtcaagacca atgcggagca
    16861 tatacgcccg gagccttggc gatcctgcaa gctccggatg cctccgctcg aagtagcgcg
    16921 tctgctgctc catacaagcc aaccacggcc tccagaagaa gatgttggcg acctcgtatt
    16981 gggaatcccc gaacatcgcc tcgctccagt caatgaccgc tgttatgcgg ccattgtccg
    17041 tcaggacatt gttggagccg aaatccgcgt gcacgaggtg ccggacttcg gggcagtcct
    17101 cggcccaaag catcagctca tcgagagcct gcgcgacgga cgcactgacg gtgtcgtcca
    17161 tcacagtttg ccagtgatac acatggggat cagcaatcgc gcatatgaaa tcacgccatg
    17221 tagtgtattg accgattcct tgcggtccga atgggccgaa cccgctcgtc tggctaagat
    17281 cggccgcagc gatagcatcc atagcctccg cgaccggctg aagaacagcg ggcagttcgg
    17341 tttcaggcag gtcttgcaac gtgacaccct gtgcacggcg ggagatgcaa taggtcaggc
    17401 tctcgctgaa ctccccaatg tcaagcactt ccggaatcgg gagcgcggcc gatgcaaagt
    17461 gccgataaac ataacgatct ttgtagaaac catcggcgca gctatttacc cgcaggacat
    17521 atccacgccc tcctacatcg aagctgaaag cacgagattc ttcgccctcc gagagctgca
    17581 tcaggtcgga gacgctgtcg aacttttcga tcagaaactt ctcgacagac gtcgcggtga
    17641 gttcaggctt tttcataggg gggattcgag ttgagagtga atatgagact ctaattggat
    17701 accgagggga atttatggaa cgtcagtgga gcatttttga caagaaatat ttgctagctg
    17761 atagtgacct taggcgactt ttgaacgcgc aataatggtt tctgacgtat gtgcttagct
    17821 cattaaactc cagaaacccg cggctcagtg gctccttcaa cgttgcggtt ctgtcagttc
    17881 caaacgtaaa acggcttgtc ccgcgtcatc ggcgggggtc ataacgtgac tcccttaatt
    17941 ctccgctcat gatcgataat tcgcggtacc cggggatcct ctagagggcc
//
