LOCUS       sequence-152377-       15443 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For CRISPR/Cas9 mediated gene editing in Arabidopsis; provides a 
            visual screen for Cas9-free plants.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15443)
  AUTHORS   Gao X, Chen J, Dai X, Zhang D, Zhao Y
  TITLE     An effective strategy for reliably isolating heritable and Cas9-free
            Arabidopsis mutants generated by CRISPR/Cas9-mediated genome 
            editing.
  JOURNAL   Plant Physiol. 2016 May 15. pii: pp.00663.2016.
  PUBMED    27208253
REFERENCE   2  (bases 1 to 15443)
  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 
            Physiol. 2016 May 15. pii: pp.00663.2016."
COMMENT     Sequence Label: pHDE-35S-Cas9-mCherry
FEATURES             Location/Qualifiers
     source          1..15443
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..25
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     900..919
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      1203..1455
                     /label=MAS terminator
                     /note="mannopine synthase terminator"
     promoter        1927..2106
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        1959..2142
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     terminator      3257..3509
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        3642..3987
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     3930..3950
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             3992..8095
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             3992..8095
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             3992..8095
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             8108..8128
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(8640..8657)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8640..8656)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8649..8671)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    8878..8902
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             9430..10221
                     /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      10465..11053
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10954..10973
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11207..11224
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    11239..11379
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(11305..11324)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     11465..11487
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      11723..11917
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(11983..13050)
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     CDS             complement(13484..14113)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
ORIGIN
        1 gtttacccgc caatatatcc tgtcaaacac tgatagttta aactagtgaa gtgaagcttg
       61 gtctcaacta tgagacctaa gctggcacaa ctatatttcc aacatcacta gctaccatca
      121 aaagattgac ttctcatctt actcgattga aaccaaatta acatagggtt tttatttaaa
      181 taaaagttta accttctttt taaaaaattg ttcatagtgt catgtcagaa caagagctac
      241 aaatcacaca tagcatgcat aagcggagct atgatgagtg gtattgtttt gttcgtcact
      301 tgtcactctt ttccaacaca taatcccgac aacaacgtaa gagcatctct ctctctccac
      361 acacactcat gcatgcatgc attcttacac gtgattgcca tgcaaatctc ctttctcacc
      421 tataaataca aaccaaccct tcactacact cttcactcaa accaaaacaa gaaaacatac
      481 acaaatagca aaacggtacc aacaatggat aacatggcca tcatcaagga gttcatgcgc
      541 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
      601 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc
      661 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg
      721 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
      781 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
      841 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
      901 cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag
      961 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac
     1021 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
     1081 aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag
     1141 tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtgagag
     1201 ctcttggact cccatgttgg caaaggcaac caaacaaaca atgaatgatc cgctcctgca
     1261 tatggggcgg tttgagtatt tcaactgcca tttgggctga attgaagaca tgctcctgtc
     1321 agaaattccg tgatcttact caatattcag taatctcggc caatatccta aatgtgcgtg
     1381 gctttatctg tctttgtatt gtttcatcaa ttcatgtaac gtttgctttt cttatgaatt
     1441 ttcaaataaa ttatcgtatt gagaccgaat tctgtctaga cctaactgag taagctagct
     1501 tgttcgagta ttatggcatt gggaaaactg tttttcttgt accatttgtt gtgcttgtaa
     1561 tttactgtgt tttttattcg gttttcgcta tcgaactgtg aaatggaaat ggatggagaa
     1621 gagttaatga atgatatggt ccttttgttc attctcaaat taatattatt tgttttttct
     1681 cttatttgtt gtgtgttgaa tttgaaatta taagagatat gcaaacattt tgttttgagt
     1741 aaaaatgtgt caaatcgtgg cctctaatga ccgaagttaa tatgaggagt aaaacactag
     1801 atccccaaac aagcttgaaa ctgaaggcgg gaaacgacaa tctgatcatg agcggagaat
     1861 taagggagtc acgttatgac ccccgccgat gacgcgggac aagccgtttt acgtttggaa
     1921 ctgacagaac cgcaacgttg aaggagccac tcagccgcgg gtttctggag tttaatgagc
     1981 taagcacata cgtcagaaac cattattgcg cgttcaaaag tcgcctaagg tcactatcag
     2041 ctagcaaata tttcttgtca aaaatgctcc actgacgttc cataaattcc cctcggtatc
     2101 caattagagt ctcatattca ctctcaatcc aaataatctg caccggatcc gctagaggat
     2161 ctcgacctgc aagatcccgg ggggcaatga gatatgaaaa agcctgaact caccgcgacg
     2221 tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg
     2281 gagggcgaag aatctcgtgc tttcagcttc gatataggag ggcgtggata tgtcctgcgg
     2341 gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcg
     2401 gccgcgctcc cgattccgga agtgcttgac attggggaat tcagcgagag cctgacctat
     2461 tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc
     2521 gctgttctgc agccggtcgc ggaggccatg gatgcgatcg ctgcggccga tcttagccag
     2581 acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat
     2641 ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc
     2701 gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc
     2761 gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc
     2821 cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc
     2881 gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc
     2941 gagcggaggc atccggagct tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt
     3001 ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg
     3061 cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc
     3121 gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga
     3181 aaccgacgcc ccagcactcg tccgggatct tggaggtgat gtaacatgat cacaagctga
     3241 tcccccgaat ttccccgatc gttcaaacat ttggcaataa agtttcttaa gattgaatcc
     3301 tgttgccggt cttgcgatga ttatcatata atttctgttg aattacgtta agcatgtaat
     3361 aattaacatg taatgcatga cgttatttat gagatgggtt tttatgatta gagtcccgca
     3421 attatacatt taatacgcga tagaaaacaa aatatagcgc gcaaactagg ataaattatc
     3481 gcgcgcggtg tcatctatgt tactagatcg ggaattgatc ccccctcgac agcttgcatg
     3541 ccagcttggg ctgcaggtcg acgcgtcaac atggtggagc acgacactct cgtctactcc
     3601 aagaatatca aagatacagt ctcagaagac caaagggcta ttgagacttt tcaacaaagg
     3661 gtaatatcgg gaaacctcct cggattccat tgcccagcta tctgtcactt catcaaaagg
     3721 acagtagaaa aggaaggtgg cacctacaaa tgccatcatt gcgataaagg aaaggctatc
     3781 gttcaagatg cctctgccga cagtggtccc aaagatggac ccccacccac gaggagcatc
     3841 gtggaaaaag aagacgttcc aaccacgtct tcaaagcaag tggattgatg tgatatctcc
     3901 actgacgtaa gggatgacgc acaatcccac tatccttcgc aagacccttc ctctatataa
     3961 ggaagttcat ttcatttgga gaggacacga tatggacaag aagtactcca ttgggctcga
     4021 tatcggcaca aacagcgtcg gctgggccgt cattacggac gagtacaagg tgccgagcaa
     4081 aaaattcaaa gttctgggca ataccgatcg ccacagcata aagaagaacc tcattggcgc
     4141 cctcctgttc gactccgggg agacggccga agccacgcgg ctcaaaagaa cagcacggcg
     4201 cagatatacc cgcagaaaga atcggatctg ctacctgcag gagatcttta gtaatgagat
     4261 ggctaaggtg gatgactctt tcttccatag gctggaggag tcctttttgg tggaggagga
     4321 taaaaagcac gagcgccacc caatctttgg caatatcgtg gacgaggtgg cgtaccatga
     4381 aaagtaccca accatatatc atctgaggaa gaagcttgta gacagtactg ataaggctga
     4441 cttgcggttg atctatctcg cgctggcgca tatgatcaaa tttcggggac acttcctcat
     4501 cgagggggac ctgaacccag acaacagcga tgtcgacaaa ctctttatcc aactggttca
     4561 gacttacaat cagcttttcg aagagaaccc gatcaacgca tccggagttg acgccaaagc
     4621 aatcctgagc gctaggctgt ccaaatcccg gcggctcgaa aacctcatcg cacagctccc
     4681 tggggagaag aagaacggcc tgtttggtaa tcttatcgcc ctgtcactcg ggctgacccc
     4741 caactttaaa tctaacttcg acctggccga agatgccaag cttcaactga gcaaagacac
     4801 ctacgatgat gatctcgaca atctgctggc ccagatcggc gaccagtacg cagacctttt
     4861 tttggcggca aagaacctgt cagacgccat tctgctgagt gatattctgc gagtgaacac
     4921 ggagatcacc aaagctccgc tgagcgctag tatgatcaag cgctatgatg agcaccacca
     4981 agacttgact ttgctgaagg cccttgtcag acagcaactg cctgagaagt acaaggaaat
     5041 tttcttcgat cagtctaaaa atggctacgc cggatacatt gacggcggag caagccagga
     5101 ggaattttac aaatttatta agcccatctt ggaaaaaatg gacggcaccg aggagctgct
     5161 ggtaaagctt aacagagaag atctgttgcg caaacagcgc actttcgaca atggaagcat
     5221 cccccaccag attcacctgg gcgaactgca cgctatcctc aggcggcaag aggatttcta
     5281 cccctttttg aaagataaca gggaaaagat tgagaaaatc ctcacatttc ggatacccta
     5341 ctatgtaggc cccctcgccc ggggaaattc cagattcgcg tggatgactc gcaaatcaga
     5401 agagaccatc actccctgga acttcgagga agtcgtggat aagggggcct ctgcccagtc
     5461 cttcatcgaa aggatgacta actttgataa aaatctgcct aacgaaaagg tgcttcctaa
     5521 acactctctg ctgtacgagt acttcacagt ttataacgag ctcaccaagg tcaaatacgt
     5581 cacagaaggg atgagaaagc cagcattcct gtctggagag cagaagaaag ctatcgtgga
     5641 cctcctcttc aagacgaacc ggaaagttac cgtgaaacag ctcaaagaag actatttcaa
     5701 aaagattgaa tgtttcgact ctgttgaaat cagcggagtg gaggatcgct tcaacgcatc
     5761 cctgggaacg tatcacgatc tcctgaaaat cattaaagac aaggacttcc tggacaatga
     5821 ggagaacgag gacattcttg aggacattgt cctcaccctt acgttgtttg aagataggga
     5881 gatgattgaa gaacgcttga aaacttacgc tcatctcttc gacgacaaag tcatgaaaca
     5941 gctcaagagg cgccgatata caggatgggg gcggctgtca agaaaactga tcaatgggat
     6001 ccgagacaag cagagtggaa agacaatcct ggattttctt aagtccgatg gatttgccaa
     6061 ccggaacttc atgcagttga tccatgatga ctctctcacc tttaaggagg acatccagaa
     6121 agcacaagtt tctggccagg gggacagtct tcacgagcac atcgctaatc ttgcaggtag
     6181 cccagctatc aaaaagggaa tactgcagac cgttaaggtc gtggatgaac tcgtcaaagt
     6241 aatgggaagg cataagcccg agaatatcgt tatcgagatg gcccgagaga accaaactac
     6301 ccagaaggga cagaagaaca gtagggaaag gatgaagagg attgaagagg gtataaaaga
     6361 actggggtcc caaatcctta aggaacaccc agttgaaaac acccagcttc agaatgagaa
     6421 gctctacctg tactacctgc agaacggcag ggacatgtac gtggatcagg aactggacat
     6481 caatcggctc tccgactacg acgtggatca tatcgtgccc cagtcttttc tcaaagatga
     6541 ttctattgat aataaagtgt tgacaagatc cgataaaaat agagggaaga gtgataacgt
     6601 cccctcagaa gaagttgtca agaaaatgaa aaattattgg cggcagctgc tgaacgccaa
     6661 actgatcaca caacggaagt tcgataatct gactaaggct gaacgaggtg gcctgtctga
     6721 gttggataaa gccggcttca tcaaaaggca gcttgttgag acacgccaga tcaccaagca
     6781 cgtggcccaa attctcgatt cacgcatgaa caccaagtac gatgaaaatg acaaactgat
     6841 tcgagaggtg aaagttatta ctctgaagtc taagctggtc tcagatttca gaaaggactt
     6901 tcagttttat aaggtgagag agatcaacaa ttaccaccat gcgcatgatg cctacctgaa
     6961 tgcagtggta ggcactgcac ttatcaaaaa atatcccaag cttgaatctg aatttgttta
     7021 cggagactat aaagtgtacg atgttaggaa aatgatcgca aagtctgagc aggaaatagg
     7081 caaggccacc gctaagtact tcttttacag caatattatg aattttttca agaccgagat
     7141 tacactggcc aatggagaga ttcggaagcg accacttatc gaaacaaacg gagaaacagg
     7201 agaaatcgtg tgggacaagg gtagggattt cgcgacagtc cggaaggtcc tgtccatgcc
     7261 gcaggtgaac atcgttaaaa agaccgaagt acagaccgga ggcttctcca aggaaagtat
     7321 cctcccgaaa aggaacagcg acaagctgat cgcacgcaaa aaagattggg accccaagaa
     7381 atacggcgga ttcgattctc ctacagtcgc ttacagtgta ctggttgtgg ccaaagtgga
     7441 gaaagggaag tctaaaaaac tcaaaagcgt caaggaactg ctgggcatca caatcatgga
     7501 gcgatcaagc ttcgaaaaaa accccatcga ctttctcgag gcgaaaggat ataaagaggt
     7561 caaaaaagac ctcatcatta agcttcccaa gtactctctc tttgagcttg aaaacggccg
     7621 gaaacgaatg ctcgctagtg cgggcgagct gcagaaaggt aacgagctgg cactgccctc
     7681 taaatacgtt aatttcttgt atctggccag ccactatgaa aagctcaaag ggtctcccga
     7741 agataatgag cagaagcagc tgttcgtgga acaacacaaa cactaccttg atgagatcat
     7801 cgagcaaata agcgaattct ccaaaagagt gatcctcgcc gacgctaacc tcgataaggt
     7861 gctttctgct tacaataagc acagggataa gcccatcagg gagcaggcag aaaacattat
     7921 ccacttgttt actctgacca acttgggcgc gcctgcagcc ttcaagtact tcgacaccac
     7981 catagacaga aagcggtaca cctctacaaa ggaggtcctg gacgccacac tgattcatca
     8041 gtcaattacg gggctctatg aaacaagaat cgacctctct cagctcggtg gagacagcag
     8101 ggctgacccc aagaagaaga ggaaggtgtg atcgatccag gcctcccagc tttcgtccgt
     8161 atcatcggtt tcgacaacgt tcgtcaagtt caatgcatca gtttcattgc ccacacacca
     8221 gaatcctact aagtttgagt attatggcat tggaaaagct gttttcttct atcatttgtt
     8281 ctgcttgtaa tttactgtgt tctttcagtt tttgttttcg gacatcaaaa tgcaaatgga
     8341 tggataagag ttaataaatg atatggtcct tttgttcatt ctcaaattat tattatctgt
     8401 tgtttttact ttaatgggtt gaatttaagt aagaaaggaa ctaacagtgt gatattaagg
     8461 tgcaatgtta gacatataaa acagtctttc acctctcttt ggttatgtct tgaattggtt
     8521 tgtttcttca cttatctgtg taatcaagtt tactatgagt ctatgatcaa gtaattatgc
     8581 aatcaagtta agtacagtat aggctttttg tgtcgagggg gtaccgagtc gaggaattca
     8641 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     8701 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     8761 ccttcccaac agttgcgcag cctgaatggc gggtaccgag ctcgaattca attcggcgtt
     8821 aattcagtac attaaaaacg tccgcaatgt gttattaagt tgtctaagcg tcaatttgtt
     8881 tacaccacaa tatatcctgc caccagccag ccaacagctc cccgaccggc agctcggcac
     8941 aaaatcacca ctcgatacag gcagcccatc agtccgggac ggcgtcagcg ggagagccgt
     9001 tgtaaggcgg cagactttgc tcatgttacc gatgctattc ggaagaacgg caactaagct
     9061 gccgggtttg aaacacggat gatctcgcgg agggtagcat gttgattgta acgatgacag
     9121 agcgttgctg cctgtgatca attcgggcac gaacccagtg gacataagcc tcgttcggtt
     9181 cgtaagctgt aatgcaagta gcgtaactgc cgtcacgcaa ctggtccaga accttgaccg
     9241 aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc ttcttgttat gacatgtttt
     9301 tttggggtac agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc cgtgggtcga
     9361 tgtttgatgt tatggagcag caacgatgtt acgcagcagg gcagtcgccc taaaacaaag
     9421 ttaaacatca tgggggaagc ggtgatcgcc gaagtatcga ctcaactatc agaggtagtt
     9481 ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg tacatttgta cggctccgca
     9541 gtggatggcg gcctgaagcc acacagtgat attgatttgc tggttacggt gaccgtaagg
     9601 cttgatgaaa caacgcggcg agctttgatc aacgaccttt tggaaacttc ggcttcccct
     9661 ggagagagcg agattctccg cgctgtagaa gtcaccattg ttgtgcacga cgacatcatt
     9721 ccgtggcgtt atccagctaa gcgcgaactg caatttggag aatggcagcg caatgacatt
     9781 cttgcaggta tcttcgagcc agccacgatc gacattgatc tggctatctt gctgacaaaa
     9841 gcaagagaac atagcgttgc cttggtaggt ccagcggcgg aggaactctt tgatccggtt
     9901 cctgaacagg atctatttga ggcgctaaat gaaaccttaa cgctatggaa ctcgccgccc
     9961 gactgggctg gcgatgagcg aaatgtagtg cttacgttgt cccgcatttg gtacagcgca
    10021 gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact gggcaatgga gcgcctgccg
    10081 gcccagtatc agcccgtcat acttgaagct agacaggctt atcttggaca agaagaagat
    10141 cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc actacgtgaa aggcgagatc
    10201 accaaggtag tcggcaaata atgtctagct agaaattcgt tcaagccgac gccgcttcgc
    10261 cggcgttaac tcaagcgatt agatgcacta agcacataat tgctcacagc caaactatca
    10321 ggtcaagtct gcttttatta tttttaagcg tgcataataa gccctacaca aattgggaga
    10381 tatatcatgc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
    10441 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
    10501 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
    10561 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
    10621 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
    10681 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
    10741 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
    10801 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
    10861 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
    10921 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
    10981 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
    11041 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
    11101 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
    11161 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
    11221 cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
    11281 acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
    11341 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
    11401 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
    11461 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
    11521 ggtgccttga tgtgggcgcc ggcggtcgag tggcgacggc gcggcttgtc cgcgccctgg
    11581 tagattgcct ggccgtaggc cagccatttt tgagcggcca gcggccgcga taggccgacg
    11641 cgaagcggcg gggcgtaggg agcgcagcga ccgaagggta ggcgcttttt gcagctcttc
    11701 ggctgtgcgc tggccagaca gttatgcaca ggccaggcgg gttttaagag ttttaataag
    11761 ttttaaagag ttttaggcgg aaaaatcgcc ttttttctct tttatatcag tcacttacat
    11821 gtgtgaccgg ttcccaatgt acggctttgg gttcccaatg tacgggttcc ggttcccaat
    11881 gtacggcttt gggttcccaa tgtacgtgct atccacagga aagagacctt ttcgaccttt
    11941 ttcccctgct agggcaattt gccctagcat ctgctccgta cattaggaac cggcggatgc
    12001 ttcgccctcg atcaggttgc ggtagcgcat gactaggatc gggccagcct gccccgcctc
    12061 ctccttcaaa tcgtactccg gcaggtcatt tgacccgatc agcttgcgca cggtgaaaca
    12121 gaacttcttg aactctccgg cgctgccact gcgttcgtag atcgtcttga acaaccatct
    12181 ggcttctgcc ttgcctgcgg cgcggcgtgc caggcggtag agaaaacggc cgatgccggg
    12241 atcgatcaaa aagtaatcgg ggtgaaccgt cagcacgtcc gggttcttgc cttctgtgat
    12301 ctcgcggtac atccaatcag ctagctcgat ctcgatgtac tccggccgcc cggtttcgct
    12361 ctttacgatc ttgtagcggc taatcaaggc ttcaccctcg gataccgtca ccaggcggcc
    12421 gttcttggcc ttcttcgtac gctgcatggc aacgtgcgtg gtgtttaacc gaatgcaggt
    12481 ttctaccagg tcgtctttct gctttccgcc atcggctcgc cggcagaact tgagtacgtc
    12541 cgcaacgtgt ggacggaaca cgcggccggg cttgtctccc ttcccttccc ggtatcggtt
    12601 catggattcg gttagatggg aaaccgccat cagtaccagg tcgtaatccc acacactggc
    12661 catgccggcc ggccctgcgg aaacctctac gtgcccgtct ggaagctcgt agcggatcac
    12721 ctcgccagct cgtcggtcac gcttcgacag acggaaaacg gccacgtcca tgatgctgcg
    12781 actatcgcgg gtgcccacgt catagagcat cggaacgaaa aaatctggtt gctcgtcgcc
    12841 cttgggcggc ttcctaatcg acggcgcacc ggctgccggc ggttgccggg attctttgcg
    12901 gattcgatca gcggccgctt gccacgattc accggggcgt gcttctgcct cgatgcgttg
    12961 ccgctgggcg gcctgcgcgg ccttcaactt ctccaccagg tcatcaccca gcgccgcgcc
    13021 gatttgtacc gggccggatg gtttgcgacc gtcacgccga ttcctcgggc ttgggggttc
    13081 cagtgccatt gcagggccgg cagacaaccc agccgcttac gcctggccaa ccgcccgttc
    13141 ctccacacat ggggcattcc acggcgtcgg tgcctggttg ttcttgattt tccatgccgc
    13201 ctcctttagc cgctaaaatt catctactca tttattcatt tgctcattta ctctggtagc
    13261 tgcgcgatgt attcagatag cagctcggta atggtcttgc cttggcgtac cgcgtacatc
    13321 ttcagcttgg tgtgatcctc cgccggcaac tgaaagttga cccgcttcat ggctggcgtg
    13381 tctgccaggc tggccaacgt tgcagccttg ctgctgcgtg cgctcggacg gccggcactt
    13441 agcgtgtttg tgcttttgct cattttctct ttacctcatt aactcaaatg agttttgatt
    13501 taatttcagc ggccagcgcc tggacctcgc gggcagcgtc gccctcgggt tctgattcaa
    13561 gaacggttgt gccggcggcg gcagtgcctg ggtagctcac gcgctgcgtg atacgggact
    13621 caagaatggg cagctcgtac ccggccagcg cctcggcaac ctcaccgccg atgcgcgtgc
    13681 ctttgatcgc ccgcgacacg acaaaggccg cttgtagcct tccatccgtg acctcaatgc
    13741 gctgcttaac cagctccacc aggtcggcgg tggcccatat gtcgtaaggg cttggctgca
    13801 ccggaatcag cacgaagtcg gctgccttga tcgcggacac agccaagtcc gccgcctggg
    13861 gcgctccgtc gatcactacg aagtcgcgcc ggccgatggc cttcacgtcg cggtcaatcg
    13921 tcgggcggtc gatgccgaca acggttagcg gttgatcttc ccgcacggcc gcccaatcgc
    13981 gggcactgcc ctggggatcg gaatcgacta acagaacatc ggccccggcg agttgcaggg
    14041 cgcgggctag atgggttgcg atggtcgtct tgcctgaccc gcctttctgg ttaagtacag
    14101 cgataacctt catgcgttcc ccttgcgtat ttgtttattt actcatcgca tcatatacgc
    14161 agcgaccgca tgacgcaagc tgttttactc aaatacacat caccttttta gacggcggcg
    14221 ctcggtttct tcagcggcca agctggccgg ccaggccgcc agcttggcat cagacaaacc
    14281 ggccaggatt tcatgcagcc gcacggttga gacgtgcgcg ggcggctcga acacgtaccc
    14341 ggccgcgatc atctccgcct cgatctcttc ggtaatgaaa aacggttcgt cctggccgtc
    14401 ctggtgcggt ttcatgcttg ttcctcttgg cgttcattct cggcggccgc cagggcgtcg
    14461 gcctcggtca atgcgtcctc acggaaggca ccgcgccgcc tggcctcggt gggcgtcact
    14521 tcctcgctgc gctcaagtgc gcggtacagg gtcgagcgat gcacgccaag cagtgcagcc
    14581 gcctctttca cggtgcggcc ttcctggtcg atcagctcgc gggcgtgcgc gatctgtgcc
    14641 ggggtgaggg tagggcgggg gccaaacttc acgcctcggg ccttggcggc ctcgcgcccg
    14701 ctccgggtgc ggtcgatgat tagggaacgc tcgaactcgg caatgccggc gaacacggtc
    14761 aacaccatgc ggccggccgg cgtggtggtg tcggcccacg gctctgccag gctacgcagg
    14821 cccgcgccgg cctcctggat gcgctcggca atgtccagta ggtcgcgggt gctgcgggcc
    14881 aggcggtcta gcctggtcac tgtcacaacg tcgccagggc gtaggtggtc aagcatcctg
    14941 gccagctccg ggcggtcgcg cctggtgccg gtgatcttct cggaaaacag cttggtgcag
    15001 ccggccgcgt gcagttcggc ccgttggttg gtcaagtcct ggtcgtcggt gctgacgcgg
    15061 gcatagccca gcaggccagc ggcggcgctc ttgttcatgg cgtaatgtct ccggttctag
    15121 tcgcaagtat tctactttat gcgactaaaa cacgcgacaa gaaaacgcca ggaaaagggc
    15181 agggcggcag cctgtcgcgt aacttaggac ttgtgcgaca tgtcgttttc agaagacggc
    15241 tgcactgaac gtcagaagcc gactgcacta tagcagcgga ggggttggat caaagtactt
    15301 taaagtactt taaagtactt taaagtactt tgatcccgag gggaaccctg tggttggcat
    15361 gcacatacaa atggacgaac ggataaacct tttcacgccc ttttaaatat ccgattattc
    15421 taataaacgc tcttttctct tag
//
