LOCUS       sequence-275276-       17545 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid with NOP1-gRNA to tranform Mp as CRISPR control. Contains 
            Cas9 and HygR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17545)
  AUTHORS   Sauret-Gueto S, Frangedakis E, Silvestri L, Rebmann M, Tomaselli M, 
            Markel K, Delmans M, West A, Patron NJ, Haseloff J
  TITLE     Systematic tools for reprogramming plant gene expression in a simple
            model, Marchantia polymorpha.
  JOURNAL   ACS Synth Biol. 2020 Mar 12. doi: 10.1021/acssynbio.9b00511.
  PUBMED    32163700
REFERENCE   2  (bases 1 to 17545)
  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: "ACS Synth 
            Biol. 2020 Mar 12. doi: 10.1021/acssynbio.9b00511."
COMMENT     Sequence Label: L2_NOP1gRNA-Cas9-CsA
FEATURES             Location/Qualifiers
     source          1..17545
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        1456..1531
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(1543..1567)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3317..7420
                     /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             3317..7420
                     /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             7433..7453
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      7477..7729
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     polyA_signal    7776..7950
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    7976..8000
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             9301..9930
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             10359..11432
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      11498..11692
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(11928..11950)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    12036..12176
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     12091..12110
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(12191..12208)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12362..12950)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12442..12461)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(13038..13829)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     misc_feature    14615..14639
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        14761..15437
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     15374..15394
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             15510..16535
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     polyA_signal    16570..16744
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
ORIGIN
        1 cttcaacttg atgaatcgga ttgatcataa aatttcacaa acttatgtgc tgattaataa
       61 aaaatataat cagatttttc atgaataatt aagataaata acacatgcaa gtctcaatta
      121 gaatttaaca atctgactat aaaaagtcgg tttatcttga ttttcacaaa aatttaacat
      181 agtttcaagg tgaatgggct tttattttaa tccaatcaac tatttgacac atacatcttg
      241 caaatttatt tattttttat ggaaaaaaac atgatttatg tttttctttg agaaagtata
      301 ttgtggattg tcaaaaaaaa cctatggaag atttatatgt aagagttaaa aatttatttt
      361 caaatatgtc aacaatccac aatatcgaag aaaattttca aaaaatcatc acatatgatg
      421 taaatataat atgaatatgt gcaatcttat ttcattttac tctgaaaact taatttcttt
      481 ttatagagca ttctgtactt actggtgtca aatatacaga taaaatgcag tccataacgt
      541 atcactctat aagcaaaatc ataaacttcg tgatctctca cagttcaaac gagaactata
      601 gaaacccttc aactagcaaa aaactcacaa ctttctattc caatatcttt aattgtctga
      661 cgatttacaa atagaaacga aaaccagaca cacacacaca cgcaaactac aaagtaaaaa
      721 cacaaccacc tgacattaga cagtgagatt attaaacatc ctaaactaaa aaacaaacaa
      781 attaattctc tatgtacaag tccccatcgt gatcaatttt tgcatcccaa tatagtggcg
      841 gttagtataa gaggaccttc gtctattcac tctccatgtt tgtcaaacta tagtacagta
      901 catatatgta tgggaacaaa ttccgaacac tctagtacac tcctcaacgc aagttctatc
      961 tcatatctct gcaagagctt attccaagaa actttcttgt gcttctatgc atgcccaggt
     1021 accttaagtc gcgcgcatag tcacttcaca gccttttcag ttctcccttt gatggctcag
     1081 agtgacgcaa ctgcacaaaa ataagaaaaa agtcgaagcg tcacgagaga tgtatccggc
     1141 tagtttcctt ggagttcgga ttgctctctt tctctgtctc cccgaaccat atggaattta
     1201 tgaactgctg ttcactagtc agtcacttgt ttcagaactt ttgagtggtc gatttatcca
     1261 ccttcttgtt tcttcctggg tccagaatct gacttcttga gacaacaaac agtgcacagt
     1321 ggatcgaaga atgaccacag ttgagacacg tggcgctggc ctcatttatt gcaccatacg
     1381 tttagtgaag aactgaataa tggtggatat aaagcagagt tgcacccagc ctctcgatag
     1441 tctttgtgag agaatgtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta
     1501 tcaacttgaa aaagtggcac cgagtcggtg ctttttttct agacccagct ttcttgtaca
     1561 aagttggccg ctacagagga gcaaatgagt cacacacatt gttgagagac atatcaaaat
     1621 aactcaattg taaggtaagt tgatagattt cccagttaaa tgcattattt taacaatatt
     1681 tgtatctcaa attttcaaaa tcatatcaaa tttgcagatc ttgatgatga ttcacaactc
     1741 tgtcttcgat ttttgattgt taacgggtat cgatatttgg gtaacttaaa aagaatttgc
     1801 taaagatctt aaaaaacaga ttaaaaaaac ctttcaaaat agctaatcct ttaaatccgt
     1861 ttcacaatat aatgctcatt gatgataatc gaaaagattc cattttaaag tgcatataaa
     1921 ttgcttaatt atctctttct gtctcgtaat attttgttct ctcctattgg atcggacccg
     1981 catcttctcg atcagttcac tatactaata tttggtcaac ttcaagagag attagaactt
     2041 cgttctccaa agtgtaaagg ccgacaagca gaaaaggaaa cttcaatcag aagccttcta
     2101 ggttttcccg cgtcccaaaa agcgaaggga ggacggggag gacggtgagg ccagaagaag
     2161 aaagcaacag taaaatctca tgtgattgcg tgcatgtcat ggatctatgg gatgctagtc
     2221 caccgtttgg aagtatgtgc agaaggagaa aggcgcggga ggccaaaagc gacttcggcc
     2281 acctctctac tttctagaaa gtttgctcgc agcttgtaat aagcgcggaa aaggtactcg
     2341 aaaaaccgga gcaatcggtg tggtatcagc agtcgggttc gctaaaccta cgtcttctta
     2401 ttttggaatg ttaaattgaa agggcgtgcc ggccctaagc acggacgatg tgacagacgg
     2461 aggagtgtga gcaattcgga tgacgatgct ggccggcagt cacgacaaag agtccgaagg
     2521 ggcatggaca gggcaatata taaatggaaa agaggctttg aatattccta acccttcgtc
     2581 gtcggcctcc caacttgccc tcgcatgctg aattggccgc ctatctgcga atggccggcc
     2641 gccgacccta agtcgcaata acttgaacgg agccgatgca aagtccttat cgaacgctcg
     2701 ttattataaa cggggcagac ctaggtattt aattatatca aaaccctccg caatttcgct
     2761 aatcattccc taatttcttc tactttggcg tctagtgctg tggtcgcgct tgctctcaaa
     2821 gctcgctggg aggcagtgga gcgtctggct taaaatcgat cgccgcaacc ctagagttcc
     2881 tttcacttct ttctctcttg cgtggagagc atcgacatcg aggagctgct acttgtcttc
     2941 tggtatgcgc cttgtgccct cgtctctgta cgtcgttacg attgctcgat attgtgattg
     3001 tctagacctg aaacccgact ggagctttta cgtggtgtcg attggggcct cgagcgagtg
     3061 attttttagg ttcgttttag ccaagtacat cccactttgg tcagtcctgt tatctcggta
     3121 aggtgttcag ttcggtcact ggtgcggagg tgcctcgtaa gtttcctgca tgatagattt
     3181 gtctatcctt gacttgaaga ggagtcgaag gtcagcgagg ttggctgtgt gctatggacg
     3241 tcggtgctcc ttggttccgt gcttctatgg gtgttgcctc aacagtattg atgtgcctgc
     3301 agaaaggttg tcaccaatgg ataagaagta ctctatcgga ctcgatatcg gaactaactc
     3361 tgtgggatgg gctgtgatca ccgatgagta caaggtgcca tctaagaagt tcaaggttct
     3421 cggaaacacc gataggcact ctatcaagaa aaaccttatc ggtgctctcc tcttcgattc
     3481 tggtgaaact gctgaggcta ccagactcaa gagaaccgct agaagaaggt acaccagaag
     3541 aaagaacagg atctgctacc tccaagagat cttctctaac gagatggcta aagtggatga
     3601 ttcattcttc cacaggctcg aagagtcatt cctcgtggaa gaagataaga agcacgagag
     3661 gcaccctatc ttcggaaaca tcgttgatga ggtggcatac cacgagaagt accctactat
     3721 ctaccacctc agaaagaagc tcgttgattc tactgataag gctgatctca ggctcatcta
     3781 cctcgctctc gctcacatga tcaagttcag aggacacttc ctcatcgagg gtgatctcaa
     3841 ccctgataac tctgatgtgg ataagttgtt catccagctc gtgcagacct acaaccagct
     3901 tttcgaagag aaccctatca acgcttcagg tgtggatgct aaggctatcc tctctgctag
     3961 gctctctaag tcaagaaggc ttgagaacct cattgctcag ctccctggtg agaagaagaa
     4021 cggacttttc ggaaacttga tcgctctctc tctcggactc acccctaact tcaagtctaa
     4081 cttcgatctc gctgaggatg caaagctcca gctctcaaag gatacctacg atgatgatct
     4141 cgataacctc ctcgctcaga tcggagatca gtacgctgat ttgttcctcg ctgctaagaa
     4201 cctctctgat gctatcctcc tcagtgatat cctcagagtg aacaccgaga tcaccaaggc
     4261 tccactctca gcttctatga tcaagagata cgatgagcac caccaggatc tcacacttct
     4321 caaggctctt gttagacagc agctcccaga gaagtacaaa gagattttct tcgatcagtc
     4381 taagaacgga tacgctggtt acatcgatgg tggtgcatct caagaagagt tctacaagtt
     4441 catcaagcct atcctcgaga agatggatgg aaccgaggaa ctcctcgtga agctcaatag
     4501 agaggatctt ctcagaaagc agaggacctt cgataacgga tctatccctc atcagatcca
     4561 cctcggagag ttgcacgcta tccttagaag gcaagaggat ttctacccat tcctcaagga
     4621 taacagggaa aagattgaga agattctcac cttcagaatc ccttactacg tgggacctct
     4681 cgctagagga aactcaagat tcgcttggat gaccagaaag tctgaggaaa ccatcacccc
     4741 ttggaacttc gaagaggtgg tggataaggg tgctagtgct cagtctttca tcgagaggat
     4801 gaccaacttc gataagaacc ttccaaacga gaaggtgctc cctaagcact ctttgctcta
     4861 cgagtacttc accgtgtaca acgagttgac caaggttaag tacgtgaccg agggaatgag
     4921 gaagcctgct tttttgtcag gtgagcaaaa gaaggctatc gttgatctct tgttcaagac
     4981 caacagaaag gtgaccgtga agcagctcaa agaggattac ttcaagaaaa tcgagtgctt
     5041 cgattcagtt gagatttctg gtgttgagga taggttcaac gcatctctcg gaacctacca
     5101 cgatctcctc aagatcatta aggataagga tttcttggat aacgaggaaa acgaggatat
     5161 cttggaggat atcgttctta ccctcaccct ctttgaagat agagagatga ttgaagaaag
     5221 gctcaagacc tacgctcatc tcttcgatga taaggtgatg aagcagttga agagaagaag
     5281 atacactggt tggggaaggc tctcaagaaa gctcattaac ggaatcaggg ataagcagtc
     5341 tggaaagaca atccttgatt tcctcaagtc tgatggattc gctaacagaa acttcatgca
     5401 gctcatccac gatgattctc tcacctttaa agaggatatc cagaaggctc aggtttcagg
     5461 acagggtgat agtctccatg agcatatcgc taacctcgct ggatctcctg caatcaagaa
     5521 gggaatcctc cagactgtga aggttgtgga tgagttggtg aaggtgatgg gaaggcataa
     5581 gcctgagaac atcgtgatcg aaatggctag agagaaccag accactcaga agggacagaa
     5641 gaactctagg gaaaggatga agaggatcga ggaaggtatc aaagagcttg gatctcagat
     5701 cctcaaagag caccctgttg agaacactca gctccagaat gagaagctct acctctacta
     5761 cctccagaac ggaagggata tgtatgtgga tcaagagttg gatatcaaca ggctctctga
     5821 ttacgatgtt gatcatatcg tgccacagtc attcttgaag gatgattcta tcgataacaa
     5881 ggtgctcacc aggtctgata agaacagggg taagagtgat aacgtgccaa gtgaagaggt
     5941 tgtgaagaaa atgaagaact attggaggca gctcctcaac gctaagctca tcactcagag
     6001 aaagttcgat aacttgacta aggctgagag gggaggactc tctgaattgg ataaggcagg
     6061 attcatcaag aggcagcttg tggaaaccag gcagatcact aagcacgttg cacagatcct
     6121 cgattctagg atgaacacca agtacgatga gaacgataag ttgatcaggg aagtgaaggt
     6181 tatcaccctc aagtcaaagc tcgtgtctga tttcagaaag gatttccaat tctacaaggt
     6241 gagggaaatc aacaactacc accacgctca cgatgcttac cttaacgctg ttgttggaac
     6301 cgctctcatc aagaagtatc ctaagctcga gtcagagttc gtgtacggtg attacaaggt
     6361 gtacgatgtg aggaagatga tcgctaagtc tgagcaagag atcggaaagg ctaccgctaa
     6421 gtatttcttc tactctaaca tcatgaattt cttcaagacc gagattaccc tcgctaacgg
     6481 tgagatcaga aagaggccac tcatcgagac aaacggtgaa acaggtgaga tcgtgtggga
     6541 taagggaagg gatttcgcta ccgttagaaa ggtgctctct atgccacagg tgaacatcgt
     6601 taagaaaacc gaggtgcaga ccggtggatt ctctaaagag tctatcctcc ctaagaggaa
     6661 ctctgataag ctcattgcta ggaagaagga ttgggaccct aagaaatacg gtggtttcga
     6721 ttctcctacc gtggcttact ctgttctcgt tgtggctaag gttgagaagg gaaagagtaa
     6781 gaagctcaag tctgttaagg aacttctcgg aatcactatc atggaaaggt catctttcga
     6841 gaagaaccca atcgatttcc tcgaggctaa gggatacaaa gaggttaaga aggatctcat
     6901 catcaagctc ccaaagtact cactcttcga actcgagaac ggtagaaaga ggatgctcgc
     6961 ttctgctggt gagcttcaaa agggaaacga gcttgctctc ccatctaagt acgttaactt
     7021 tctttacctc gcttctcact acgagaagtt gaagggatct ccagaagata acgagcagaa
     7081 gcaacttttc gttgagcagc acaagcacta cttggatgag atcatcgagc agatctctga
     7141 gttctctaaa agggtgatcc tcgctgatgc aaacctcgat aaggtgttgt ctgcttacaa
     7201 caagcacaga gataagccta tcagggaaca ggcagagaac atcatccatc tcttcaccct
     7261 taccaacctc ggtgctcctg ctgctttcaa gtacttcgat acaaccatcg ataggaagag
     7321 atacacctct accaaagaag tgctcgatgc taccctcatc catcagtcta tcactggact
     7381 ctacgagact aggatcgatc tctcacagct cggtggtgat tcaagggctg atcctaagaa
     7441 gaagaggaag gtttgagctt gagctcgaat ttccccgatc gttcaaacat ttggcaataa
     7501 agtttcttaa gattgaatcc tgttgccggt cttgcgatga ttatcatata atttctgttg
     7561 aattacgtta agcatgtaat aattaacatg taatgcatga cgttatttat gagatgggtt
     7621 tttatgatta gagtcccgca attatacatt taatacgcga tagaaaacaa aatatagcgc
     7681 gcaaactagg ataaattatc gcgcgcggtg tcatctatgt tactagatcg atccgtatcg
     7741 atagcctcta gctagagtcg atcgacaagc tcgagtttct ccataataat gtgtgagtag
     7801 ttcccagata agggaattag ggttcctata gggtttcgct catgtgttga gcatataaga
     7861 aacccttagt atgtatttgt atttgtaaaa tacttctatc aataaaattt ctaattccta
     7921 aaatcaaaat ccagtactaa aatccagatc gctaggtata cttgagaccg gatcctgaca
     7981 ggatatattg gcgggtaaac ctaagagaaa agagcgttta ttagaataat cggatattta
     8041 aaagggcgtg aaaaggttta tccgttcgtc catttgtatg tgcatgccaa ccacagggtt
     8101 cccctcggga tcaaagtact ttgatccaac ccctccgctg ctatagtgca gtcggcttct
     8161 gacgttcagt gcagccgtca tctgaaaacg acatgtcgca caagtcctaa gttacgcgac
     8221 aggctgccgc cctgcccttt tcctggcgtt ttcttgtcgc gtgttttagt cgcataaagt
     8281 agaatacttg cgactagaac cggagacatt acgccatgaa caagagcgcc gccgctggcc
     8341 tgctgggcta tgcccgcgtc agcaccgacg accaggactt gaccaaccaa cgggccgaac
     8401 tgcacgcggc cggctgcacc aagctgtttt ccgagaagat caccggcacc aggcgcgacc
     8461 gcccggagct ggccaggatg cttgaccacc tacgccctgg cgacgttgtg acagtgacca
     8521 ggctagaccg cctggcccgc agcacccgcg acctactgga cattgccgag cgcatccagg
     8581 aggccggcgc gggcctgcgt agcctggcag agccgtgggc cgacaccacc acgccggccg
     8641 gccgcatggt gttgaccgtg ttcgccggca ttgccgagtt cgagcgttcc ctaatcatcg
     8701 accgcacccg gagcgggcgc gaggccgcca aggcccgagg cgtgaagttt ggcccccgcc
     8761 ctaccctcac cccggcacag atcgcgcacg cccgcgagct gatcgaccag gaaggccgca
     8821 ccgtgaaaga ggcggctgca ctgcttggcg tgcatcgctc gaccctgtac cgcgcacttg
     8881 agcgcagcga ggaagtgacg cccaccgagg ccaggcggcg cggtgccttc cgtgaggacg
     8941 cattgaccga ggccgacgcc ctggcggccg ccgagaatga acgccaagag gaacaagcat
     9001 gaaaccgcac caggacggcc aggacgaacc gtttttcatt accgaagaga tcgaggcgga
     9061 gatgatcgcg gccgggtacg tgttcgagcc gcccgcgcac ctctcaaccg tgcggctgca
     9121 tgaaatcctg gccggtttgt ctgatgccaa gctggcggcc tggccggcca gcttggccgc
     9181 tgaagaaacc gagcgccgcc gtctaaaaag gtgatgtgta tttgagtaaa acagcttgcg
     9241 tcatgcggtc gctgcgtata tgatgcgatg agtaaataaa caaatacgca aggggaacgc
     9301 atgaaggtta tcgctgtact taaccagaaa ggcgggtcag gcaagacgac catcgcaacc
     9361 catctagccc gcgccctgca actcgccggg gccgatgttc tgttagtcga ttccgatccc
     9421 cagggcagtg cccgcgattg ggcggccgtg cgggaagatc aaccgctaac cgttgtcggc
     9481 atcgaccgcc cgacgattga ccgcgacgtg aaggccatcg gccggcgcga cttcgtagtg
     9541 atcgacggag cgccccaggc ggcggacttg gctgtgtccg cgatcaaggc agccgacttc
     9601 gtgctgattc cggtgcagcc aagcccttac gacatatggg ccaccgccga cctggtggag
     9661 ctggttaagc agcgcattga ggtcacggat ggaaggctac aagcggcctt tgtcgtgtcg
     9721 cgggcgatca aaggcacgcg catcggcggt gaggttgccg aggcgctggc cgggtacgag
     9781 ctgcccattc ttgagtcccg tatcacgcag cgcgtgagct acccaggcac tgccgccgcc
     9841 ggcacaaccg ttcttgaatc agaacccgag ggcgacgctg cccgcgaggt ccaggcgctg
     9901 gccgctgaaa ttaaatcaaa actcatttga gttaatgagg taaagagaaa atgagcaaaa
     9961 gcacaaacac gctaagtgcc ggccgtccga gcgcacgcag cagcaaggct gcaacgttgg
    10021 ccagcctggc agacacgcca gccatgaagc gggtcaactt tcagttgccg gcggaggatc
    10081 acaccaagct gaagatgtac gcggtacgcc aaggcaagac cattaccgag ctgctatctg
    10141 aatacatcgc gcagctacca gagtaaatga gcaaatgaat aaatgagtag atgaatttta
    10201 gcggctaaag gaggcggcat ggaaaatcaa gaacaaccag gcaccgacgc cgtggaatgc
    10261 cccatgtgtg gaggaacggg cggttggcca ggcgtaagcg gctgggttgc ctgccggccc
    10321 tgcaatggca ctggaacccc caagcccgag gaatcggcgt gagcggtcgc aaaccatccg
    10381 gcccggtaca aatcggcgcg gcgctgggtg atgacctggt ggagaagttg aaggccgcgc
    10441 aggccgccca gcggcaacgc atcgaggcag aagcacgccc cggtgaatcg tggcaagcgg
    10501 ccgctgatcg aatccgcaaa gaatcccggc aaccgccggc agccggtgcg ccgtcgatta
    10561 ggaagccgcc caagggcgac gagcaaccag attttttcgt tccgatgctc tatgacgtgg
    10621 gcacccgcga tagtcgcagc atcatggacg tggccgtttt ccgtctgtcg aagcgtgacc
    10681 gacgagctgg cgaggtgatc cgctacgagc ttccagacgg gcacgtagag gtttccgcag
    10741 ggccggccgg catggccagt gtgtgggatt acgacctggt actgatggcg gtttcccatc
    10801 taaccgaatc catgaaccga taccgggaag ggaagggaga caagcccggc cgcgtgttcc
    10861 gtccacacgt tgcggacgta ctcaagttct gccggcgagc cgatggcgga aagcagaaag
    10921 acgacctggt agaaacctgc attcggttaa acaccacgca cgttgccatg cagcgtacga
    10981 agaaggccaa gaacggccgc ctggtgacgg tatccgaggg tgaagccttg attagccgct
    11041 acaagatcgt aaagagcgaa accgggcggc cggagtacat cgagatcgag ctagctgatt
    11101 ggatgtaccg cgagatcaca gaaggcaaga acccggacgt gctgacggtt caccccgatt
    11161 actttttgat cgatcccggc atcggccgtt ttctctaccg cctggcacgc cgcgccgcag
    11221 gcaaggcaga agccagatgg ttgttcaaga cgatctacga acgcagtggc agcgccggag
    11281 agttcaagaa gttctgtttc accgtgcgca agctgatcgg gtcaaatgac ctgccggagt
    11341 acgatttgaa ggaggaggcg gggcaggctg gcccgatcct agtcatgcgc taccgcaacc
    11401 tgatcgaggg cgaagcatcc gccggttcct aatgtacgga gcagatgcta gggcaaattg
    11461 ccctagcagg ggaaaaaggt cgaaaaggac tctttcctgt ggatagcacg tacattggga
    11521 acccaaagcc gtacattggg aaccggaacc cgtacattgg gaacccaaag ccgtacattg
    11581 ggaaccggtc acacatgtaa gtgactgata taaaagagaa aaaaggcgat ttttccgcct
    11641 aaaactcttt aaaacttatt aaaactctta aaacccgcct ggcctgtgca taactgtctg
    11701 gccagcgcac agccgaagtg ctgcaaaaag cgcctaccct tcggtcgctg cgctccctac
    11761 gccccgccgc ttcgcgtcgg cctatcgcgg ccgctggccg ctcaaaaatg gctggcctac
    11821 ggccaggcaa tctaccaggg cgcggacaag ccgcgccgtc gccactcgac cgccggcgcc
    11881 cacatcaagg caccctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
    11941 cagctcccgg tgacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
    12001 cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc
    12061 gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
    12121 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct
    12181 catccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
    12241 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
    12301 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
    12361 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
    12421 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
    12481 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
    12541 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
    12601 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
    12661 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
    12721 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
    12781 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
    12841 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
    12901 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
    12961 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
    13021 tcatgcattc taggtgatta tttgccgact accttggtga tctcgccttt cacgtagtgg
    13081 acaaattctt ccaactgatc tgcgcgcgag gccaagcgat cttcttcttg tccaagataa
    13141 gcctgtctag cttcaagtat gacgggctga tactgggccg gcaggcgctc cattgcccag
    13201 tcggcagcga catccttcgg cgcgattttg ccggttactg cgctgtacca aatgcgggac
    13261 aacgtaagca ctacatttcg ctcatcacca gcccagtcgg gcggcgagtt ccatagcgtt
    13321 aaggtttcat ttagcgcctc aaatagatcc tgttcaggaa ccggatcaaa gagttcctcc
    13381 gccgctggac ctaccaaggc aacgctatgt tctcttgctt ttgtcagcaa gatagccaga
    13441 tcaatgtcga tcgtggctgg ctcgaagata cctgcaagaa tgtcattgcg ctgccattct
    13501 ccaaattgca gttcgcgctt agctggataa cgccacggaa tgatgtcgtc gtgcacaaca
    13561 atggtgactt ctacagcgcg gagaatctcg ctctctccag gggaagccga agtttccaaa
    13621 aggtcgttga tcaaagctcg ccgcgttgtt tcatcaagcc ttacggtcac cgtaaccagc
    13681 aaatcaatat cactgtgtgg cttcaggccg ccatccactg cggagccgta caaatgtacg
    13741 gccagcaacg tcggttcgag atggcgctcg atgacgccaa ctacctctga tagttgagtc
    13801 gatacttcgg cgatcaccgc ttccctcata atgtttaact ttgttttagg gcgactgccc
    13861 tgctgcgtaa catcgttgct gctccataac atcaaacatc gacccacggc gtaacgcgct
    13921 tgctgcttgg atgcccgagg catagactgt accccaaaaa aacagtcata acaagccatg
    13981 aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca aggttctgga ccagttgcgt
    14041 gagcgcatac gctacttgca ttacagctta cgaaccgaac aggcttatgt ccactgggtt
    14101 cgtgccttca tccgtttcca cggtgtgcgt cacccggcaa ccttgggtag cagcgaagtc
    14161 gaggcatttc tgtcctggct ggaacagaac ttattatttc cttcctcttt tctacagtat
    14221 ttaaagatac cccaagaagc taattataac aagacgaact ccaattcact gttccttgca
    14281 ttctaaaacc ttaaatacca gaaaacagct ttttcaaagt tgttttcaaa gttggcgtat
    14341 aacatagtat cgacggagcc gattttgaaa ccgcggtgat cacaggcagc aacgctctgt
    14401 catcgttaca atcaacatgc taccctccgc gagatcatcc gtgtttcaaa cccggcagct
    14461 tagttgccgt tcttccgaat agcatcggta acatgagcaa agtctgccgc cttacaacgg
    14521 ctctcccgct gacgccgtcc cggactgatg ggctgcctgt atcgagtggt gattttgtgc
    14581 cgagctgccg gtcggggagc tgttggctgg ctggtggcag gatatattgt ggtgtaaaca
    14641 taacggatcc ggtctcagga gaatgaggag gctagagcag cttgccaaca tggtggagca
    14701 cgacactctc gtctactcca agaatatcaa agatacagtc tcagaagacc aaagggctat
    14761 tgagactttt caacaaaggg taatatcggg aaacctcctc ggattccatt gcccagctat
    14821 ctgtcacttc atcaaaagga cagtagaaaa ggaaggtggc acctacaaat gccatcattg
    14881 cgataaagga aaggctatcg ttcaagatgc ctctgccgac agtggtccca aagatggacc
    14941 cccacccacg aggagcatcg tggaaaaaga agacgttcca accacgtctt caaagcaagt
    15001 ggattgatgt gaacatggtg gagcacgaca ctctcgtcta ctccaagaat atcaaagata
    15061 cagtctcaga agaccaaagg gctattgaga cttttcaaca aagggtaata tcgggaaacc
    15121 tcctcggatt ccattgccca gctatctgtc acttcatcaa aaggacagta gaaaaggaag
    15181 gtggcaccta caaatgccat cattgcgata aaggaaaggc tatcgttcaa gatgcctctg
    15241 ccgacagtgg tcccaaagat ggacccccac ccacgaggag catcgtggaa aaagaagacg
    15301 ttccaaccac gtcttcaaag caagtggatt gatgtgatat ctccactgac gtaagggatg
    15361 acgcacaatc ccactatcct tcgcaagacc cttcctctat ataaggaagt tcatttcatt
    15421 tggagaggac acgctgaaat caccagtctc tctctacaaa tctatctctc tcgagctttc
    15481 gcagatccgg ggggcaatga gattcaccaa tgaaaaagcc tgaactcacc gcgacgtctg
    15541 tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga cctgatgcag ctctcggagg
    15601 gcgaagaatc tcgtgctttc agcttcgatg taggagggcg tggatatgtc ctgcgggtaa
    15661 atagctgcgc cgatggtttc tacaaagatc gttatgttta tcggcacttt gcatcggccg
    15721 cgctcccgat tccggaagtg cttgacattg gggagtttag cgagagcctg acctattgca
    15781 tctcccgccg ttcacagggt gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg
    15841 ttctacaacc ggtcgcggag gctatggatg cgatcgctgc ggccgatctt agccagacga
    15901 gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata cactacatgg cgtgatttca
    15961 tatgcgcgat tgctgatccc catgtgtatc actggcaaac tgtgatggac gacaccgtca
    16021 gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac tgccccgaag
    16081 tccggcacct cgtgcacgcg gatttcggct ccaacaatgt cctgacggac aatggccgca
    16141 taacagcggt cattgactgg agcgaggcga tgttcgggga ttcccaatac gaggtcgcca
    16201 acatcttctt ctggaggccg tggttggctt gtatggagca gcagacgcgc tacttcgagc
    16261 ggaggcatcc ggagcttgca ggatcgccac gactccgggc gtatatgctc cgcattggtc
    16321 ttgaccaact ctatcagagc ttggttgacg gcaatttcga tgatgcagct tgggcgcagg
    16381 gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca caaatcgccc
    16441 gcagaagcgc ggccgtctgg accgatggct gtgtagaagt actcgccgat agtggaaacc
    16501 gacgccccag cactcgtccg agggcaaaga aataggcttc tctagctaga gtcgatcgac
    16561 aagctcgagt ttctccataa taatgtgtga gtagttccca gataagggaa ttagggttcc
    16621 tatagggttt cgctcatgtg ttgagcatat aagaaaccct tagtatgtat ttgtatttgt
    16681 aaaatacttc tatcaataaa atttctaatt cctaaaacca aaatccagta ctaaaatcca
    16741 gatcgctagc aaggagtctg agcaggacaa ctcgcgtagt gagagttaca tgttcgttgg
    16801 gttcttccga cacggacctg agttggccaa cgtcccacct gaggtctgtg ccccggtgat
    16861 gagaagtgtg catctcgttc ttgcagctcg tcagtacttt cagaatcatg gcgtgcatgg
    16921 tagaatgacc cttataacgg acttcgacat ggcaatcgct atacaggagt attcattcaa
    16981 aagagatttt taaagatcaa tttttctttc tcataagtct aatacgtcat gataataact
    17041 ttaaaaaaaa ttgaactcgt taaaaaattt gatcaaattt taatttatta ttagatcata
    17101 tattacatat cagtttcata atgttatact aagatctttc atatatattt taacccacga
    17161 gacaaaatat gtttatatct ttatttattt aaatattttt tagctcaaaa catcgcaaac
    17221 tcagctatca taatgctttt taatgaaaaa aacaaatttt cctatcatac tctcattgtt
    17281 tataattttt tagaaaaaaa acttgaagat aaaataacaa caataatagt ttcaaaagca
    17341 catttttgtt gtaaatttca agaaaaatac aatcatatta ctgattgact acaaaaattt
    17401 gtaaattaat ccaggctaca tgtttaagat gtgtcaaaac tgcatcatct acatctcatt
    17461 ttaatttttc taaactacat acatagtatc aataacaatt gaacaatgct agttaatgtg
    17521 ttatactcat tagcttgtaa tatat
//
