LOCUS       sequence-211624-       15937 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cas9 expression in M.polymorpha.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15937)
  AUTHORS   Sugano SS, Nishihama R, Shirakawa M, Takagi J, Matsuda Y, Ishida S, 
            Shimada T, Hara-Nishimura I, Osakabe K, Kohchi T
  TITLE     Efficient CRISPR/Cas9-based genome editing and its application to 
            conditional genetic analysis in Marchantia polymorpha.
  JOURNAL   PLoS One. 2018 Oct 31;13(10):e0205117. doi: 
            10.1371/journal.pone.0205117. eCollection 2018.
  PUBMED    30379827
REFERENCE   2  (bases 1 to 15937)
  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: "PLoS One.";
            date: "2018-10-31"; volume: "13(10):e0205117. doi"; pages: " 
            10.1371/journal.pone.0205117. eCollection 2018"
COMMENT     Sequence Label: pMpGE006
FEATURES             Location/Qualifiers
     source          1..15937
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1743..1767
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1788..5891
                     /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             1788..5891
                     /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             5904..5924
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(6427..6451)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      6483..6735
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(6768..6784)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6768..6784)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6781..6803)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6792..6808
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6816..6822,6823..6840,6841..6846))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6861..6882
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     polyA_signal    6967..7141
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(7182..8207)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     promoter        complement(8274..8950)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     complement(8317..8337)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     misc_feature    9145..9169
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             9690..10481
                     /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      10727..11315
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11216..11235
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11469..11486
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    11501..11641
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(11567..11586)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     11727..11749
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      11985..12179
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(12245..13318)
                     /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(13747..14376)
                     /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    15677..15701
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     15889..15911
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     15903..15920
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     15904..15920
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaatgagtc acacacattg ttgagagaca tatcaaaata actcaattgt aaggtaagtt
       61 gatagatttc ccagttaaat gcattatttt aacaatattt gtatctcaaa ttttcaaaat
      121 catatcaaat ttgcagatct tgatgatgat tcacaactct gtcttcgatt tttgattgtt
      181 aacgggtatc gatatttggg taacttaaaa agaatttgct aaagatctta aaaaacagat
      241 taaaaaaacc tttcaaaata gctaatcctt taaatccgtt tcacaatata atgctcattg
      301 atgataatcg aaaagattcc attttaaagt gcatataaat tgcttaatta tctctttctg
      361 tctcgtaata ttttgttctc tcctattgga tcggacccgc atcttctcga tcagttcact
      421 atactaatat ttggtcaact tcaagagaga ttagaacttc gttctccaaa gtgtaaaggc
      481 cgacaagcag aaaaggaaac ttcaatcaga agccttctag gttttcccgc gtcccaaaaa
      541 gcgaagggag gacggggagg acggtgaggc cagaagaaga aagcaacagt aaaatctcat
      601 gtgattgcgt gcatgtcatg gatctatggg atgctagtcc accgtttgga agtatgtgca
      661 gaaggagaaa ggcgcgggag gccaaaagcg acttcggcca cctctctact ttctagaaag
      721 tttgctcgca gcttgtaata agcgcggaaa aggtactcga aaaaccggag caatcggtgt
      781 ggtatcagca gtcgggttcg ctaaacctac gtcttcttat tttggaatgt taaattgaaa
      841 gggcgtgccg gccctaagca cggacgatgt gacagacgga ggagtgtgag caattcggat
      901 gacgatgctg gccggcagtc acgacaaaga gtccgaaggg gcatggacag ggcaatatat
      961 aaatggaaaa gaggctttga atattcctaa cccttcgtcg tcggcctccc aacttgccct
     1021 cgcatgctga attggccgcc tatctgcgaa tggccggccg ccgaccctaa gtcgcaataa
     1081 cttgaacgga gccgatgcaa agtccttatc gaacgctcgt tattataaac ggggcagacc
     1141 taggtattta attatatcaa aaccctccgc aatttcgcta atcattccct aatttcttct
     1201 actttggcgt ctagtgctgt ggtcgcgctt gctctcaaag ctcgctggga ggcagtggag
     1261 cgtctggctt aaaatcgatc gccgcaaccc tagagttcct ttcacttctt tctctcttgc
     1321 gtggagagca tcgacatcga agagctgcat cttgtcttct ggtatgcgcc ttgtgccctc
     1381 gtctctgtac gtcgttacga ttgctcgata ttgtgattgt ctagacctga aacccgactg
     1441 gagcttttac gtggtgtcga ttggggcctc gagcgagtga ttttttaggt tcgttttagc
     1501 caagtacatc ccactttggt cagtcctgtt atctcggtaa ggtgttcagt tcggtctctg
     1561 gtgcggaggt gcctcgtaag tttcctgcat gatagatttg tctatccttg acttgaagag
     1621 gagtcgaagg tcagcgaggt tggctgtgtg ctatggacgt cggtgctcct tggttccgtg
     1681 cttctatggg tgttgcctca acagtattga tgtgcctgca gaaaggttga ctagagttat
     1741 caacaagttt gtacaaaaaa gcaggctccg cggccgcccc cttcaccatg gataagaagt
     1801 actctatcgg actcgatatc ggaactaact ctgtgggatg ggctgtgatc accgatgagt
     1861 acaaggtgcc atctaagaag ttcaaggttc tcggaaacac cgataggcac tctatcaaga
     1921 aaaaccttat cggtgctctc ctcttcgatt ctggtgaaac tgctgaggct accagactca
     1981 agagaaccgc tagaagaagg tacaccagaa gaaagaacag gatctgctac ctccaagaga
     2041 tcttctctaa cgagatggct aaagtggatg attcattctt ccacaggctc gaagagtcat
     2101 tcctcgtgga agaagataag aagcacgaga ggcaccctat cttcggaaac atcgttgatg
     2161 aggtggcata ccacgagaag taccctacta tctaccacct cagaaagaag ctcgttgatt
     2221 ctactgataa ggctgatctc aggctcatct acctcgctct cgctcacatg atcaagttca
     2281 gaggacactt cctcatcgag ggtgatctca accctgataa ctctgatgtg gataagttgt
     2341 tcatccagct cgtgcagacc tacaaccagc ttttcgaaga gaaccctatc aacgcttcag
     2401 gtgtggatgc taaggctatc ctctctgcta ggctctctaa gtcaagaagg cttgagaacc
     2461 tcattgctca gctccctggt gagaagaaga acggactttt cggaaacttg atcgctctct
     2521 ctctcggact cacccctaac ttcaagtcta acttcgatct cgctgaggat gcaaagctcc
     2581 agctctcaaa ggatacctac gatgatgatc tcgataacct cctcgctcag atcggagatc
     2641 agtacgctga tttgttcctc gctgctaaga acctctctga tgctatcctc ctcagtgata
     2701 tcctcagagt gaacaccgag atcaccaagg ctccactctc agcttctatg atcaagagat
     2761 acgatgagca ccaccaggat ctcacacttc tcaaggctct tgttagacag cagctcccag
     2821 agaagtacaa agagattttc ttcgatcagt ctaagaacgg atacgctggt tacatcgatg
     2881 gtggtgcatc tcaagaagag ttctacaagt tcatcaagcc tatcctcgag aagatggatg
     2941 gaaccgagga actcctcgtg aagctcaata gagaggatct tctcagaaag cagaggacct
     3001 tcgataacgg atctatccct catcagatcc acctcggaga gttgcacgct atccttagaa
     3061 ggcaagagga tttctaccca ttcctcaagg ataacaggga aaagattgag aagattctca
     3121 ccttcagaat cccttactac gtgggacctc tcgctagagg aaactcaaga ttcgcttgga
     3181 tgaccagaaa gtctgaggaa accatcaccc cttggaactt cgaagaggtg gtggataagg
     3241 gtgctagtgc tcagtctttc atcgagagga tgaccaactt cgataagaac cttccaaacg
     3301 agaaggtgct ccctaagcac tctttgctct acgagtactt caccgtgtac aacgagttga
     3361 ccaaggttaa gtacgtgacc gagggaatga ggaagcctgc ttttttgtca ggtgagcaaa
     3421 agaaggctat cgttgatctc ttgttcaaga ccaacagaaa ggtgaccgtg aagcagctca
     3481 aagaggatta cttcaagaaa atcgagtgct tcgattcagt tgagatttct ggtgttgagg
     3541 ataggttcaa cgcatctctc ggaacctacc acgatctcct caagatcatt aaggataagg
     3601 atttcttgga taacgaggaa aacgaggata tcttggagga tatcgttctt accctcaccc
     3661 tctttgaaga tagagagatg attgaagaaa ggctcaagac ctacgctcat ctcttcgatg
     3721 ataaggtgat gaagcagttg aagagaagaa gatacactgg ttggggaagg ctctcaagaa
     3781 agctcattaa cggaatcagg gataagcagt ctggaaagac aatccttgat ttcctcaagt
     3841 ctgatggatt cgctaacaga aacttcatgc agctcatcca cgatgattct ctcaccttta
     3901 aagaggatat ccagaaggct caggtttcag gacagggtga tagtctccat gagcatatcg
     3961 ctaacctcgc tggatctcct gcaatcaaga agggaatcct ccagactgtg aaggttgtgg
     4021 atgagttggt gaaggtgatg ggaaggcata agcctgagaa catcgtgatc gaaatggcta
     4081 gagagaacca gaccactcag aagggacaga agaactctag ggaaaggatg aagaggatcg
     4141 aggaaggtat caaagagctt ggatctcaga tcctcaaaga gcaccctgtt gagaacactc
     4201 agctccagaa tgagaagctc tacctctact acctccagaa cggaagggat atgtatgtgg
     4261 atcaagagtt ggatatcaac aggctctctg attacgatgt tgatcatatc gtgccacagt
     4321 cattcttgaa ggatgattct atcgataaca aggtgctcac caggtctgat aagaacaggg
     4381 gtaagagtga taacgtgcca agtgaagagg ttgtgaagaa aatgaagaac tattggaggc
     4441 agctcctcaa cgctaagctc atcactcaga gaaagttcga taacttgact aaggctgaga
     4501 ggggaggact ctctgaattg gataaggcag gattcatcaa gaggcagctt gtggaaacca
     4561 ggcagatcac taagcacgtt gcacagatcc tcgattctag gatgaacacc aagtacgatg
     4621 agaacgataa gttgatcagg gaagtgaagg ttatcaccct caagtcaaag ctcgtgtctg
     4681 atttcagaaa ggatttccaa ttctacaagg tgagggaaat caacaactac caccacgctc
     4741 acgatgctta ccttaacgct gttgttggaa ccgctctcat caagaagtat cctaagctcg
     4801 agtcagagtt cgtgtacggt gattacaagg tgtacgatgt gaggaagatg atcgctaagt
     4861 ctgagcaaga gatcggaaag gctaccgcta agtatttctt ctactctaac atcatgaatt
     4921 tcttcaagac cgagattacc ctcgctaacg gtgagatcag aaagaggcca ctcatcgaga
     4981 caaacggtga aacaggtgag atcgtgtggg ataagggaag ggatttcgct accgttagaa
     5041 aggtgctctc tatgccacag gtgaacatcg ttaagaaaac cgaggtgcag accggtggat
     5101 tctctaaaga gtctatcctc cctaagagga actctgataa gctcattgct aggaagaagg
     5161 attgggaccc taagaaatac ggtggtttcg attctcctac cgtggcttac tctgttctcg
     5221 ttgtggctaa ggttgagaag ggaaagagta agaagctcaa gtctgttaag gaacttctcg
     5281 gaatcactat catggaaagg tcatctttcg agaagaaccc aatcgatttc ctcgaggcta
     5341 agggatacaa agaggttaag aaggatctca tcatcaagct cccaaagtac tcactcttcg
     5401 aactcgagaa cggtagaaag aggatgctcg cttctgctgg tgagcttcaa aagggaaacg
     5461 agcttgctct cccatctaag tacgttaact ttctttacct cgcttctcac tacgagaagt
     5521 tgaagggatc tccagaagat aacgagcaga agcaactttt cgttgagcag cacaagcact
     5581 acttggatga gatcatcgag cagatctctg agttctctaa aagggtgatc ctcgctgatg
     5641 caaacctcga taaggtgttg tctgcttaca acaagcacag agataagcct atcagggaac
     5701 aggcagagaa catcatccat ctcttcaccc ttaccaacct cggtgctcct gctgctttca
     5761 agtacttcga tacaaccatc gataggaaga gatacacctc taccaaagaa gtgctcgatg
     5821 ctaccctcat ccatcagtct atcactggac tctacgagac taggatcgat ctctcacagc
     5881 tcggtggtga ttcaagggct gatcctaaga agaagaggaa ggtttgaggc gcgccgagct
     5941 ccaggcctcc cagctttcgt ccgtatcatc ggtttcgaca acgttcgtca agttcaatgc
     6001 atcagtttca ttgcccacac accagaatcc tactaagttt gagtattatg gcattggaaa
     6061 agctgttttc ttctatcatt tgttctgctt gtaatttact gtgttctttc agtttttgtt
     6121 ttcggacatc aaaatgcaaa tggatggata agagttaata aatgatatgg tccttttgtt
     6181 cattctcaaa ttattattat ctgttgtttt tactttaatg ggttgaattt aagtaagaaa
     6241 ggaactaaca gtgtgatatt aaggtgcaat gttagacata taaaacagtc tttcacctct
     6301 ctttggttat gtcttgaatt ggtttgtttc ttcacttatc tgtgtaatca agtttactat
     6361 gagtctatga tcaagtaatt atgcaatcaa gttaagtaca gtataggctt aagggtgggc
     6421 gcgccgaccc agctttcttg tacaaagtgg ttgataacag cgcttagagc tcgaatttcc
     6481 ccgatcgttc aaacatttgg caataaagtt tcttaagatt gaatcctgtt gccggtcttg
     6541 cgatgattat catataattt ctgttgaatt acgttaagca tgtaataatt aacatgtaat
     6601 gcatgacgtt atttatgaga tgggttttta tgattagagt cccgcaatta tacatttaat
     6661 acgcgataga aaacaaaata tagcgcgcaa actaggataa attatcgcgc gcggtgtcat
     6721 ctatgttact agatcgggaa ttggttccgg aaccaattcg taatcatgtc atagctgttt
     6781 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     6841 tgtaaagcct ggggtgccta atgagtgagc taactcacat taggctgaat taggcgcgcc
     6901 tatttctgaa ttcaacacat tgcggacgtt tttaatgtac tgaattaacg ccgaattaat
     6961 tcgggggatc tggattttag tactggattt tggttttagg aattagaaat tttattgata
     7021 gaagtatttt acaaatacaa atacatacta agggtttctt atatgctcaa cacatgagcg
     7081 aaaccctata ggaaccctaa ttcccttatc tgggaactac tcacacatta ttatggagaa
     7141 actcgagctt gtcgatcgac agatcccggt cggcatctac tctatttctt tgccctcgga
     7201 cgagtgctgg ggcgtcggtt tccactatcg gcgagtactt ctacacagcc atcggtccag
     7261 acggccgcgc ttctgcgggc gatttgtgta cgcccgacag tcccggctcc ggatcggacg
     7321 attgcgtcgc atcgaccctg cgcccaagct gcatcatcga aattgccgtc aaccaagctc
     7381 tgatagagtt ggtcaagacc aatgcggagc atatacgccc ggagtcgtgg cgatcctgca
     7441 agctccggat gcctccgctc gaagtagcgc gtctgctgct ccatacaagc caaccacggc
     7501 ctccagaaga agatgttggc gacctcgtat tgggaatccc cgaacatcgc ctcgctccag
     7561 tcaatgaccg ctgttatgcg gccattgtcc gtcaggacat tgttggagcc gaaatccgcg
     7621 tgcacgaggt gccggacttc ggggcagtcc tcggcccaaa gcatcagctc atcgagagcc
     7681 tgcgcgacgg acgcactgac ggtgtcgtcc atcacagttt gccagtgata cacatgggga
     7741 tcagcaatcg cgcatatgaa atcacgccat gtagtgtatt gaccgattcc ttgcggtccg
     7801 aatgggccga acccgctcgt ctggctaaga tcggccgcag cgatcgcatc catagcctcc
     7861 gcgaccggtt gtagaacagc gggcagttcg gtttcaggca ggtcttgcaa cgtgacaccc
     7921 tgtgaacggc gggagatgca ataggtcagg ctctcgctaa actccccaat gtcaagcact
     7981 tccggaatcg ggagcgcggc cgatgcaaag tgccgataaa cataacgatc tttgtagaaa
     8041 ccatcggcgc agctatttac ccgcaggaca tatccacgcc ctcctacatc gaagctgaaa
     8101 gcacgagatt cttcgccctc cgagagctgc atcaggtcgg agacgctgtc gaacttttcg
     8161 atcagaaact tctcgacaga cgtcgcggtg agttcaggct ttttcatatc tcattgcccc
     8221 ccggatctgc gaaagctcga gagagataga tttgtagaga gagactggtg atttcagcgt
     8281 gtcctctcca aatgaaatga acttccttat atagaggaag ggtcttgcga aggatagtgg
     8341 gattgtgcgt catcccttac gtcagtggag atatcacatc aatccacttg ctttgaagac
     8401 gtggttggaa cgtcttcttt ttccacgatg ctcctcgtgg gtgggggtcc atctttggga
     8461 ccactgtcgg cagaggcatc ttgaacgata gcctttcctt tatcgcaatg atggcatttg
     8521 taggtgccac cttccttttc tactgtcctt ttgatgaagt gacagatagc tgggcaatgg
     8581 aatccgagga ggtttcccga tattaccctt tgttgaaaag tctcaatagc cctttggtct
     8641 tctgagactg tatctttgat attcttggag tagacgagag tgtcgtgctc caccatgttc
     8701 acatcaatcc acttgctttg aagacgtggt tggaacgtct tctttttcca cgatgctcct
     8761 cgtgggtggg ggtccatctt tgggaccact gtcggcagag gcatcttgaa cgatagcctt
     8821 tcctttatcg caatgatggc atttgtaggt gccaccttcc ttttctactg tccttttgat
     8881 gaagtgacag atagctgggc aatggaatcc gaggaggttt cccgatatta ccctttgttg
     8941 aaaagtctca atagcccttt ggtcttctga gactgtatct ttgatattct tggagtagac
     9001 gagagtgtcg tgctccacca tgttggcaag ctgctctagc caatacgcaa accgaattca
     9061 gaaataaatt cagcctaatt cggcgttaat tcagtacatt aaaaacgtcc gcaatgtgtt
     9121 attaagttgt ctaagcgtca atttgtttac accacaatat atcctgccac cagccagcca
     9181 acagctcccc gaccggcagc tcggcacaaa atcaccactc gatacaggca gcccatcagt
     9241 ccgggacggc gtcagcggga gagccgttgt aaggcggcag actttgctca tgttaccgat
     9301 gctattcgga agaacggcaa ctaagctgcc gggtttgaaa cacggatgat ctcgcggagg
     9361 gtagcatgtt gattgtaacg atgacagagc gttgctgcct gtgatcaatt cgggcacgaa
     9421 cccagtggac ataagcctgt tcggttcgta agctgtaatg caagtagcgt atgcgctcac
     9481 gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg gcggttttca
     9541 tggcttgtta tgactgtttt tttggggtac agtctatgcc tcgggcatcc aagcagcaag
     9601 cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagg
     9661 gcagtcgccc taaaacaaag ttaaacatca tgggggaagc ggtgatcgcc gaagtatcga
     9721 ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg
     9781 tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagtgat attgatttgc
     9841 tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt
     9901 tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg
     9961 ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag
    10021 aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc
    10081 tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg
    10141 aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa
    10201 cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt
    10261 cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact
    10321 gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct agacaggctt
    10381 atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc
    10441 actacgtgaa aggcgagatc accaaggtag tcggcaaata atgtctagct agaaattcgt
    10501 tcaagccgac gccgcttcgc ggcgcggctt aactcaagcg ttagatgcac taagcacata
    10561 attgctcaca gccaaactat caggtcaagt ctgcttttat tatttttaag cgtgcataat
    10621 aagccctaca caaattggga gatatatcat gcatgaccaa aatcccttaa cgtgagtttt
    10681 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
    10741 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
    10801 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
    10861 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
    10921 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
    10981 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
    11041 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
    11101 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
    11161 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
    11221 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
    11281 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
    11341 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
    11401 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
    11461 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
    11521 ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg
    11581 atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc
    11641 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
    11701 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
    11761 atcaccgaaa cgcgcgaggc agggtgcctt gatgtgggcg ccggcggtcg agtggcgacg
    11821 gcgcggcttg tccgcgccct ggtagattgc ctggccgtag gccagccatt tttgagcggc
    11881 cagcggccgc gataggccga cgcgaagcgg cggggcgtag ggagcgcagc gaccgaaggg
    11941 taggcgcttt ttgcagctct tcggctgtgc gctggccaga cagttatgca caggccaggc
    12001 gggttttaag agttttaata agttttaaag agttttaggc ggaaaaatcg ccttttttct
    12061 cttttatatc agtcacttac atgtgtgacc ggttcccaat gtacggcttt gggttcccaa
    12121 tgtacgggtt ccggttccca atgtacggct ttgggttccc aatgtacgtg ctatccacag
    12181 gaaagagacc ttttcgacct ttttcccctg ctagggcaat ttgccctagc atctgctccg
    12241 tacattagga accggcggat gcttcgccct cgatcaggtt gcggtagcgc atgactagga
    12301 tcgggccagc ctgccccgcc tcctccttca aatcgtactc cggcaggtca tttgacccga
    12361 tcagcttgcg cacggtgaaa cagaacttct tgaactctcc ggcgctgcca ctgcgttcgt
    12421 agatcgtctt gaacaaccat ctggcttctg ccttgcctgc ggcgcggcgt gccaggcggt
    12481 agagaaaacg gccgatgccg ggatcgatca aaaagtaatc ggggtgaacc gtcagcacgt
    12541 ccgggttctt gccttctgtg atctcgcggt acatccaatc agctagctcg atctcgatgt
    12601 actccggccg cccggtttcg ctctttacga tcttgtagcg gctaatcaag gcttcaccct
    12661 cggataccgt caccaggcgg ccgttcttgg ccttcttcgt acgctgcatg gcaacgtgcg
    12721 tggtgtttaa ccgaatgcag gtttctacca ggtcgtcttt ctgctttccg ccatcggctc
    12781 gccggcagaa cttgagtacg tccgcaacgt gtggacggaa cacgcggccg ggcttgtctc
    12841 ccttcccttc ccggtatcgg ttcatggatt cggttagatg ggaaaccgcc atcagtacca
    12901 ggtcgtaatc ccacacactg gccatgccgg ccggccctgc ggaaacctct acgtgcccgt
    12961 ctggaagctc gtagcggatc acctcgccag ctcgtcggtc acgcttcgac agacggaaaa
    13021 cggccacgtc catgatgctg cgactatcgc gggtgcccac gtcatagagc atcggaacga
    13081 aaaaatctgg ttgctcgtcg cccttgggcg gcttcctaat cgacggcgca ccggctgccg
    13141 gcggttgccg ggattctttg cggattcgat cagcggccgc ttgccacgat tcaccggggc
    13201 gtgcttctgc ctcgatgcgt tgccgctggg cggcctgcgc ggccttcaac ttctccacca
    13261 ggtcatcacc cagcgccgcg ccgatttgta ccgggccgga tggtttgcga ccgctcacgc
    13321 cgattcctcg ggcttggggg ttccagtgcc attgcagggc cggcagacaa cccagccgct
    13381 tacgcctggc caaccgcccg ttcctccaca catggggcat tccacggcgt cggtgcctgg
    13441 ttgttcttga ttttccatgc cgcctccttt agccgctaaa attcatctac tcatttattc
    13501 atttgctcat ttactctggt agctgcgcga tgtattcaga tagcagctcg gtaatggtct
    13561 tgccttggcg taccgcgtac atcttcagct tggtgtgatc ctccgccggc aactgaaagt
    13621 tgacccgctt catggctggc gtgtctgcca ggctggccaa cgttgcagcc ttgctgctgc
    13681 gtgcgctcgg acggccggca cttagcgtgt ttgtgctttt gctcattttc tctttacctc
    13741 attaactcaa atgagttttg atttaatttc agcggccagc gcctggacct cgcgggcagc
    13801 gtcgccctcg ggttctgatt caagaacggt tgtgccggcg gcggcagtgc ctgggtagct
    13861 cacgcgctgc gtgatacggg actcaagaat gggcagctcg tacccggcca gcgcctcggc
    13921 aacctcaccg ccgatgcgcg tgcctttgat cgcccgcgac acgacaaagg ccgcttgtag
    13981 ccttccatcc gtgacctcaa tgcgctgctt aaccagctcc accaggtcgg cggtggccca
    14041 tatgtcgtaa gggcttggct gcaccggaat cagcacgaag tcggctgcct tgatcgcgga
    14101 cacagccaag tccgccgcct ggggcgctcc gtcgatcact acgaagtcgc gccggccgat
    14161 ggccttcacg tcgcggtcaa tcgtcgggcg gtcgatgccg acaacggtta gcggttgatc
    14221 ttcccgcacg gccgcccaat cgcgggcact gccctgggga tcggaatcga ctaacagaac
    14281 atcggccccg gcgagttgca gggcgcgggc tagatgggtt gcgatggtcg tcttgcctga
    14341 cccgcctttc tggttaagta cagcgataac cttcatgcgt tccccttgcg tatttgttta
    14401 tttactcatc gcatcatata cgcagcgacc gcatgacgca agctgtttta ctcaaataca
    14461 catcaccttt ttagacggcg gcgctcggtt tcttcagcgg ccaagctggc cggccaggcc
    14521 gccagcttgg catcagacaa accggccagg atttcatgca gccgcacggt tgagacgtgc
    14581 gcgggcggct cgaacacgta cccggccgcg atcatctccg cctcgatctc ttcggtaatg
    14641 aaaaacggtt cgtcctggcc gtcctggtgc ggtttcatgc ttgttcctct tggcgttcat
    14701 tctcggcggc cgccagggcg tcggcctcgg tcaatgcgtc ctcacggaag gcaccgcgcc
    14761 gcctggcctc ggtgggcgtc acttcctcgc tgcgctcaag tgcgcggtac agggtcgagc
    14821 gatgcacgcc aagcagtgca gccgcctctt tcacggtgcg gccttcctgg tcgatcagct
    14881 cgcgggcgtg cgcgatctgt gccggggtga gggtagggcg ggggccaaac ttcacgcctc
    14941 gggccttggc ggcctcgcgc ccgctccggg tgcggtcgat gattagggaa cgctcgaact
    15001 cggcaatgcc ggcgaacacg gtcaacacca tgcggccggc cggcgtggtg gtgtcggccc
    15061 acggctctgc caggctacgc aggcccgcgc cggcctcctg gatgcgctcg gcaatgtcca
    15121 gtaggtcgcg ggtgctgcgg gccaggcggt ctagcctggt cactgtcaca acgtcgccag
    15181 ggcgtaggtg gtcaagcatc ctggccagct ccgggcggtc gcgcctggtg ccggtgatct
    15241 tctcggaaaa cagcttggtg cagccggccg cgtgcagttc ggcccgttgg ttggtcaagt
    15301 cctggtcgtc ggtgctgacg cgggcatagc ccagcaggcc agcggcggcg ctcttgttca
    15361 tggcgtaatg tctccggttc tagtcgcaag tattctactt tatgcgacta aaacacgcga
    15421 caagaaaacg ccaggaaaag ggcagggcgg cagcctgtcg cgtaacttag gacttgtgcg
    15481 acatgtcgtt ttcagaagac ggctgcactg aacgtcagaa gccgactgca ctatagcagc
    15541 ggaggggttg gatcaaagta ctttgatccc gaggggaacc ctgtggttgg catgcacata
    15601 caaatggacg aacggataaa ccttttcacg cccttttaaa tatccgatta ttctaataaa
    15661 cgctcttttc tcttaggttt acccgccaat atatcctgtc aaacactgat agtttaaact
    15721 gaaggcggga aacgacaatc tgatccaagc tcaagctgct ctagcattcg ccattcaggc
    15781 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
    15841 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
    15901 gttgtaaaac gacggccagt gccaagcttg gcccggg
//
