LOCUS       sequence-211943-       17650 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  All-in-one genome editing vector in M. polymorpha (chlorsulfuron 
            resistant).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17650)
  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 17650)
  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: pMpGE014
FEATURES             Location/Qualifiers
     source          1..17650
                     /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"
     protein_bind    6480..6504
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        7065..7140
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(7182..7206)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      7246..7498
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(7531..7547)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7531..7547)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7544..7566)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7555..7571
                     /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(7579..7585,7586..7603,7604..7609))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7624..7645
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     polyA_signal    7730..7904
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     promoter        complement(9987..10663)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     complement(10030..10050)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     misc_feature    10858..10882
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             11403..12194
                     /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      12440..13028
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12929..12948
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     13182..13199
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    13214..13354
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(13280..13299)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     13440..13462
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      13698..13892
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(13958..15031)
                     /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(15460..16089)
                     /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    17390..17414
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     17602..17624
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     17616..17633
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     17617..17633
                     /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 cggttgacta gagttatcaa
     6481 caagtttgta caaaaaagca ggctccgcgg ccgccccctt gtaacgtgag actactacac
     6541 tcctcaacgc aagttctatc tcatatctct gcaagagctt attccaagaa actttcttgt
     6601 gcttctatgc atgcccaggt accttaagtc gcgcgcatag tcacttcaca gccttttcag
     6661 ttctcccttt gatggctcag agtgacgcaa ctgcacaaaa ataagaaaaa agtcgaagcg
     6721 tcacgagaga tgtatccggc tagtttcctt ggagttcgga ttgctctctt tctctgtctc
     6781 cccgaaccat atggaattta tgaactgctg ttcactagtc agtcacttgt ttcagaactt
     6841 ttgagtggtc gatttatcca ccttcttgtt tcttcctggg tccagaatct gacttcttga
     6901 gacaacaaac agtgcacagt ggatcgaaga atgaccacag ttgagacacg tggcgctggc
     6961 ctcatttatt gcaccatacg tttagtgaag aactgaataa tggtggatat aaagcagagt
     7021 tgcacccagc ctctcgaaat gcaggtgatg actcacctgc ataagtttta gagctagaaa
     7081 tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc
     7141 tttttttcta gtatcgggaa ttctaaaggg tgggcgcgcc gacccagctt tcttgtacaa
     7201 agtggttgat aacagcgctt agaggcttag agctcgaatt tccccgatcg ttcaaacatt
     7261 tggcaataaa gtttcttaag attgaatcct gttgccggtc ttgcgatgat tatcatataa
     7321 tttctgttga attacgttaa gcatgtaata attaacatgt aatgcatgac gttatttatg
     7381 agatgggttt ttatgattag agtcccgcaa ttatacattt aatacgcgat agaaaacaaa
     7441 atatagcgcg caaactagga taaattatcg cgcgcggtgt catctatgtt actagatcgg
     7501 gaattggttc cggaaccaat tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta
     7561 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     7621 ctaatgagtg agctaactca cattaggctg aattaggcgc gcctatttct gaattcaaca
     7681 cattgcggac gtttttaatg tactgaatta acgccgaatt aattcggggg atctggattt
     7741 tagtactgga ttttggtttt aggaattaga aattttattg atagaagtat tttacaaata
     7801 caaatacata ctaagggttt cttatatgct caacacatga gcgaaaccct ataggaaccc
     7861 taattccctt atctgggaac tactcacaca ttattatgga gaaactcgag ttagtatata
     7921 gtccttccat caccctctgt gataatctcc ttaaaacttg ctcccccagg aatcataggc
     7981 agaacgtgct cctggtgggg aactatcaca tccagtaaat aaggaccggg agaatctagc
     8041 atagtttgaa ttgctgctct tagttcatcc ttcttcacca cacgcgcatt tggcaccttg
     8101 aaaccgtctg caactgtaca gaagtcgggg tagatctgct cctcgtgatc tggatcacct
     8161 aagtaagtat gtgctctgtt ggccttgtag aatctatcct ccaattgtac gaccattcct
     8221 agatgttggt tgtttaagat catgatcttc acgggtaact tctccacaaa aattgtagcc
     8281 aactcctgta cgttcatcac aaagctacca tcgccatcga tgtcgatcac aggaatatct
     8341 ggttgagttg cggcagcccc caaagcagaa ggaagtccga atcccatggc tccaagacct
     8401 ccagatgtca accaacgccg cggttcgttg tacttgaacc actgggctgc ccacatttga
     8461 tgttggccaa caccagtaga tatgatggca tttccaccag tgagttcgta cagcgtctgt
     8521 atggcagatt gaggagcaat ggcatcctcg aatacgggaa acttcattgg ccatttctgc
     8581 ttttgttctt ctagttctgc cacccagttg gagaagtcga aagtcctctt ttgactcttc
     8641 aagagggcat tgatttcact tagcgcaggt ttcacatcag cacacatgga cacatggggc
     8701 tgtttgttct ttccaatttc agcagggtca atatcgatat gaacgatgct ggacctgcta
     8761 gcaaaagtct ccagtttacc agtcactcga tcgtcgaaac gcactccaaa tgccaacaac
     8821 agatctgctt ggtccacggc gtagttggcg taaacagttc catgcattcc aagcattttc
     8881 aaacttaatg gatcagactc gggatacgct cccaaaccca ttaaagtcga tgtgatagga
     8941 attccggtaa gctcgacaaa ttccctcaac tcttcgctgg cgtgattgca acctcctcca
     9001 gaatacacta caggcttctt agaactagaa atgagtctta taatctgctc gattaacgtt
     9061 ggttccggtg gccgaggcag tcgttgtaag tagcccttca gttgcattgg ctgagaccaa
     9121 tctggaatag ccatttgctg ttgcacgtcc ttgggaatat ctaccaagac tgggcccggc
     9181 ctacctgaag ctgccaaaaa gaaagcctcc ctgattacgc gagggatgtc atcaatgttc
     9241 ataacaaggt aattgtgctt ggtgatcgat ctcgtaacct ccacaattgg agtttcttga
     9301 aaagcatcgg taccaatcat cctactggat acttgacctg taatggccac tagtggcaca
     9361 ctgtccaaga gggcatcagc gagcccggta acaagattgg tcgctccagg accggatgtg
     9421 gctatacaaa cccctactcg tccagacgat ttagcataac cttcagcagc gaaaatctcc
     9481 ccctgctcgt ggcgacagag aatgttgcgg atgctgctgg aacgtgtaag agcctggtga
     9541 atctccatag acgctccgcc gggataagcg aacaccgttc taatcccttc tcgctcaaga
     9601 gcctccacca aaatgtcggc tcctttgcgg ggctcgtcat ccttgtagcg agactgaaag
     9661 tcggacgggg ttggtgcaac gagtgacttc ccgctgctaa gccgtggtcc actcacatcg
     9721 actgcagact ggatcttctc gctaacagct tgggctttca cttccagtga acgctcagtc
     9781 ttccgagagg tgttgtatgc atgtcccctc agagcggctc ccgagaagaa accactcctc
     9841 acctttagct ctttagctcc cttgcaacaa gaccttgccg atggctgcac cttctccagg
     9901 ctcgctatgc tcaaaggcac agctgcgcat ccactaattg acgccatgct cgagagagat
     9961 agatttgtag agagagactg gtgatttcag cgtgtcctct ccaaatgaaa tgaacttcct
    10021 tatatagagg aagggtcttg cgaaggatag tgggattgtg cgtcatccct tacgtcagtg
    10081 gagatatcac atcaatccac ttgctttgaa gacgtggttg gaacgtcttc tttttccacg
    10141 atgctcctcg tgggtggggg tccatctttg ggaccactgt cggcagaggc atcttgaacg
    10201 atagcctttc ctttatcgca attatggcat ttgtaggtgc caccttcctt ttctactgtc
    10261 cttttgatga agtgacagat agctgggcaa tggaatccga ggaggtttcc cgatattacc
    10321 ctttgttgaa aagtctcaat agccctttgg tcttctgaga ctgtatcttt gatattcttg
    10381 gagtagacga gagtgtcgtg ctccaccatg ttcacatcaa tccacttgct ttgaagacgt
    10441 ggttggaacg tcttcttttt ccacgatgct cctcgtgggt gggggtccat ctttgggacc
    10501 actgtcggca gaggcatctt gaacgatagc ctttccttta tcgcaatgat ggcatttgta
    10561 ggtgccacct tccttttcta ctgtcctttt gatgaagtga cagatagctg ggcaatggaa
    10621 tccgaggagg tttcccgata ttaccctttg ttgaaaagtc tcaatagccc tttggtcttc
    10681 tgagactgta tctttgatat tcttggagta gacgagagtg tcgtgctcca ccatgttggc
    10741 aagctgctct agccaatacg caaaccgaat tcagaaataa attcagccta attcggcgtt
    10801 aattcagtac attaaaaacg tccgcaatgt gttattaagt tgtctaagcg tcaatttgtt
    10861 tacaccacaa tatatcctgc caccagccag ccaacagctc cccgaccggc agctcggcac
    10921 aaaatcacca ctcgatacag gcagcccatc agtccgggac ggcgtcagcg ggagagccgt
    10981 tgtaaggcgg cagactttgc tcatgttacc gatgctattc ggaagaacgg caactaagct
    11041 gccgggtttg aaacacggat gatctcgcgg agggtagcat gttgattgta acgatgacag
    11101 agcgttgctg cctgtgatca attcgggcac gaacccagtg gacataagcc tgttcggttc
    11161 gtaagctgta atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac
    11221 gcagcggtgg taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgggg
    11281 tacagtctat gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga
    11341 tgttatggag cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca
    11401 tcatggggga agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca
    11461 tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg
    11521 gcggcctgaa gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg
    11581 aaacaacgcg gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga
    11641 gcgagattct ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc
    11701 gttatccagc taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag
    11761 gtatcttcga gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag
    11821 aacatagcgt tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac
    11881 aggatctatt tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg
    11941 ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg
    12001 gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt
    12061 atcagcccgt catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg
    12121 cctcgcgcgc agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg
    12181 tagtcggcaa ataatgtcta gctagaaatt cgttcaagcc gacgccgctt cgcggcgcgg
    12241 cttaactcaa gcgttagatg cactaagcac ataattgctc acagccaaac tatcaggtca
    12301 agtctgcttt tattattttt aagcgtgcat aataagccct acacaaattg ggagatatat
    12361 catgcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
    12421 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
    12481 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
    12541 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
    12601 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
    12661 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
    12721 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
    12781 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
    12841 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
    12901 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
    12961 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
    13021 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
    13081 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
    13141 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
    13201 aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
    13261 gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac
    13321 tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga
    13381 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
    13441 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagggtgc
    13501 cttgatgtgg gcgccggcgg tcgagtggcg acggcgcggc ttgtccgcgc cctggtagat
    13561 tgcctggccg taggccagcc atttttgagc ggccagcggc cgcgataggc cgacgcgaag
    13621 cggcggggcg tagggagcgc agcgaccgaa gggtaggcgc tttttgcagc tcttcggctg
    13681 tgcgctggcc agacagttat gcacaggcca ggcgggtttt aagagtttta ataagtttta
    13741 aagagtttta ggcggaaaaa tcgccttttt tctcttttat atcagtcact tacatgtgtg
    13801 accggttccc aatgtacggc tttgggttcc caatgtacgg gttccggttc ccaatgtacg
    13861 gctttgggtt cccaatgtac gtgctatcca caggaaagag accttttcga cctttttccc
    13921 ctgctagggc aatttgccct agcatctgct ccgtacatta ggaaccggcg gatgcttcgc
    13981 cctcgatcag gttgcggtag cgcatgacta ggatcgggcc agcctgcccc gcctcctcct
    14041 tcaaatcgta ctccggcagg tcatttgacc cgatcagctt gcgcacggtg aaacagaact
    14101 tcttgaactc tccggcgctg ccactgcgtt cgtagatcgt cttgaacaac catctggctt
    14161 ctgccttgcc tgcggcgcgg cgtgccaggc ggtagagaaa acggccgatg ccgggatcga
    14221 tcaaaaagta atcggggtga accgtcagca cgtccgggtt cttgccttct gtgatctcgc
    14281 ggtacatcca atcagctagc tcgatctcga tgtactccgg ccgcccggtt tcgctcttta
    14341 cgatcttgta gcggctaatc aaggcttcac cctcggatac cgtcaccagg cggccgttct
    14401 tggccttctt cgtacgctgc atggcaacgt gcgtggtgtt taaccgaatg caggtttcta
    14461 ccaggtcgtc tttctgcttt ccgccatcgg ctcgccggca gaacttgagt acgtccgcaa
    14521 cgtgtggacg gaacacgcgg ccgggcttgt ctcccttccc ttcccggtat cggttcatgg
    14581 attcggttag atgggaaacc gccatcagta ccaggtcgta atcccacaca ctggccatgc
    14641 cggccggccc tgcggaaacc tctacgtgcc cgtctggaag ctcgtagcgg atcacctcgc
    14701 cagctcgtcg gtcacgcttc gacagacgga aaacggccac gtccatgatg ctgcgactat
    14761 cgcgggtgcc cacgtcatag agcatcggaa cgaaaaaatc tggttgctcg tcgcccttgg
    14821 gcggcttcct aatcgacggc gcaccggctg ccggcggttg ccgggattct ttgcggattc
    14881 gatcagcggc cgcttgccac gattcaccgg ggcgtgcttc tgcctcgatg cgttgccgct
    14941 gggcggcctg cgcggccttc aacttctcca ccaggtcatc acccagcgcc gcgccgattt
    15001 gtaccgggcc ggatggtttg cgaccgctca cgccgattcc tcgggcttgg gggttccagt
    15061 gccattgcag ggccggcaga caacccagcc gcttacgcct ggccaaccgc ccgttcctcc
    15121 acacatgggg cattccacgg cgtcggtgcc tggttgttct tgattttcca tgccgcctcc
    15181 tttagccgct aaaattcatc tactcattta ttcatttgct catttactct ggtagctgcg
    15241 cgatgtattc agatagcagc tcggtaatgg tcttgccttg gcgtaccgcg tacatcttca
    15301 gcttggtgtg atcctccgcc ggcaactgaa agttgacccg cttcatggct ggcgtgtctg
    15361 ccaggctggc caacgttgca gccttgctgc tgcgtgcgct cggacggccg gcacttagcg
    15421 tgtttgtgct tttgctcatt ttctctttac ctcattaact caaatgagtt ttgatttaat
    15481 ttcagcggcc agcgcctgga cctcgcgggc agcgtcgccc tcgggttctg attcaagaac
    15541 ggttgtgccg gcggcggcag tgcctgggta gctcacgcgc tgcgtgatac gggactcaag
    15601 aatgggcagc tcgtacccgg ccagcgcctc ggcaacctca ccgccgatgc gcgtgccttt
    15661 gatcgcccgc gacacgacaa aggccgcttg tagccttcca tccgtgacct caatgcgctg
    15721 cttaaccagc tccaccaggt cggcggtggc ccatatgtcg taagggcttg gctgcaccgg
    15781 aatcagcacg aagtcggctg ccttgatcgc ggacacagcc aagtccgccg cctggggcgc
    15841 tccgtcgatc actacgaagt cgcgccggcc gatggccttc acgtcgcggt caatcgtcgg
    15901 gcggtcgatg ccgacaacgg ttagcggttg atcttcccgc acggccgccc aatcgcgggc
    15961 actgccctgg ggatcggaat cgactaacag aacatcggcc ccggcgagtt gcagggcgcg
    16021 ggctagatgg gttgcgatgg tcgtcttgcc tgacccgcct ttctggttaa gtacagcgat
    16081 aaccttcatg cgttcccctt gcgtatttgt ttatttactc atcgcatcat atacgcagcg
    16141 accgcatgac gcaagctgtt ttactcaaat acacatcacc tttttagacg gcggcgctcg
    16201 gtttcttcag cggccaagct ggccggccag gccgccagct tggcatcaga caaaccggcc
    16261 aggatttcat gcagccgcac ggttgagacg tgcgcgggcg gctcgaacac gtacccggcc
    16321 gcgatcatct ccgcctcgat ctcttcggta atgaaaaacg gttcgtcctg gccgtcctgg
    16381 tgcggtttca tgcttgttcc tcttggcgtt cattctcggc ggccgccagg gcgtcggcct
    16441 cggtcaatgc gtcctcacgg aaggcaccgc gccgcctggc ctcggtgggc gtcacttcct
    16501 cgctgcgctc aagtgcgcgg tacagggtcg agcgatgcac gccaagcagt gcagccgcct
    16561 ctttcacggt gcggccttcc tggtcgatca gctcgcgggc gtgcgcgatc tgtgccgggg
    16621 tgagggtagg gcgggggcca aacttcacgc ctcgggcctt ggcggcctcg cgcccgctcc
    16681 gggtgcggtc gatgattagg gaacgctcga actcggcaat gccggcgaac acggtcaaca
    16741 ccatgcggcc ggccggcgtg gtggtgtcgg cccacggctc tgccaggcta cgcaggcccg
    16801 cgccggcctc ctggatgcgc tcggcaatgt ccagtaggtc gcgggtgctg cgggccaggc
    16861 ggtctagcct ggtcactgtc acaacgtcgc cagggcgtag gtggtcaagc atcctggcca
    16921 gctccgggcg gtcgcgcctg gtgccggtga tcttctcgga aaacagcttg gtgcagccgg
    16981 ccgcgtgcag ttcggcccgt tggttggtca agtcctggtc gtcggtgctg acgcgggcat
    17041 agcccagcag gccagcggcg gcgctcttgt tcatggcgta atgtctccgg ttctagtcgc
    17101 aagtattcta ctttatgcga ctaaaacacg cgacaagaaa acgccaggaa aagggcaggg
    17161 cggcagcctg tcgcgtaact taggacttgt gcgacatgtc gttttcagaa gacggctgca
    17221 ctgaacgtca gaagccgact gcactatagc agcggagggg ttggatcaaa gtactttgat
    17281 cccgagggga accctgtggt tggcatgcac atacaaatgg acgaacggat aaaccttttc
    17341 acgccctttt aaatatccga ttattctaat aaacgctctt ttctcttagg tttacccgcc
    17401 aatatatcct gtcaaacact gatagtttaa actgaaggcg ggaaacgaca atctgatcca
    17461 agctcaagct gctctagcat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
    17521 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
    17581 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc
    17641 ttggcccggg
//
