LOCUS       sequence-351574-       15392 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Binary expression vector with A.th. codon-optimized Cas9 and sgRNA 
            for miRNA390a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15392)
  AUTHORS   Lukan T, Veillet F, Kriznik M, Coll A, Mahkovec Povalej T, Pogacar 
            K, Stare K, Chauvin L, Chauvin JE, Gruden K
  TITLE     CRISPR/Cas9-mediated fine-tuning of miRNA expression in tetraploid 
            potato.
  JOURNAL   Hortic Res. 2022 Jun 30;9:uhac147. doi: 10.1093/hr/uhac147. 
            eCollection 2022.
  PUBMED    36072839
REFERENCE   2  (bases 1 to 15392)
  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: "Hortic 
            Res."; date: "2022-06-30"; volume: "9:uhac147. doi"; pages: " 
            10.1093/hr/uhac147. eCollection 2022"
COMMENT     Sequence Label: pDe_Cas9_KANA_R390a
FEATURES             Location/Qualifiers
     source          1..15392
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(164..186)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    272..412
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     327..346
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(427..444)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(598..1186)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(678..697)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1432..2223)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     misc_feature    2744..2768
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     protein_bind    2945..2966
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2981..2986,2987..3004,3005..3011)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3019..3035
                     /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)."
     primer_bind     3024..3046
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3043..3059
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3043..3059
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4060..8163
                     /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             4060..8163
                     /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             8176..8196
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    8708..8732
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      9245..9264
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     terminator      9705..9724
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     protein_bind    complement(9819..9843)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      complement(9973..10225)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(10265..11059)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(10376..10395)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     10986..11005
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(11093..11272)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     complement(11362..11379)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(11362..11378)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11371..11393)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    11574..11598
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             12929..13558
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             13987..15060
                     /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      15126..15320
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
ORIGIN
        1 cgccgcttcg cgtcggccta tcgcggccgc tggccgctca aaaatggctg gcctacggcc
       61 aggcaatcta ccagggcgcg gacaagccgc gccgtcgcca ctcgaccgcc ggcgcccaca
      121 tcaaggcacc ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
      181 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
      241 gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata
      301 gcggagtgta tactggctta actatgcggc atcagagcag attgtactga gagtgcacca
      361 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc
      421 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
      481 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
      541 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
      601 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
      661 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
      721 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
      781 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
      841 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
      901 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
      961 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     1021 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     1081 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     1141 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     1201 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     1261 gcatgatata tctcccaatt tgtgtagggc ttattatgca cgcttaaaaa taataaaagc
     1321 agacttgacc tgatagtttg gctgtgagca attatgtgct tagtgcatct aacgcttgag
     1381 ttaagccgcg ccgcgaagcg gcgtcggctt gaacgaattt ctagctagac attatttgcc
     1441 gactaccttg gtgatctcgc ctttcacgta gtggacaaat tcttccaact gatctgcgcg
     1501 cgaggccaag cgatcttctt cttgtccaag ataagcctgt ctagcttcaa gtatgacggg
     1561 ctgatactgg gccggcaggc gctccattgc ccagtcggca gcgacatcct tcggcgcgat
     1621 tttgccggtt actgcgctgt accaaatgcg ggacaacgta agcactacat ttcgctcatc
     1681 gccagcccag tcgggcggcg agttccatag cgttaaggtt tcatttagcg cctcaaatag
     1741 atcctgttca ggaaccggat caaagagttc ctccgccgct ggacctacca aggcaacgct
     1801 atgttctctt gcttttgtca gcaagatagc cagatcaatg tcgatcgtgg ctggctcgaa
     1861 gatacctgca agaatgtcat tgcgctgcca ttctccaaat tgcagttcgc gcttagctgg
     1921 ataacgccac ggaatgatgt cgtcgtgcac aacaatggtg acttctacag cgcggagaat
     1981 ctcgctctct ccaggggaag ccgaagtttc caaaaggtcg ttgatcaaag ctcgccgcgt
     2041 tgtttcatca agccttacgg tcaccgtaac cagcaaatca atatcactgt gtggcttcag
     2101 gccgccatcc actgcggagc cgtacaaatg tacggccagc aacgtcggtt cgagatggcg
     2161 ctcgatgacg ccaactacct ctgatagttg agtcgatact tcggcgatca ccgcttcccc
     2221 catgatgttt aactttgttt tagggcgact gccctgctgc gtaacatcgt tgctgctcca
     2281 taacatcaaa catcgaccca cggcgtaacg cgcttgctgc ttggatgccc gaggcataga
     2341 ctgtacccca aaaaaacagt cataacaagc catgaaaacc gccactgcgc cgttaccacc
     2401 gctgcgttcg gtcaaggttc tggaccagtt gcgtgagcgc atacgctact tgcattacag
     2461 cttacgaacc gaacaggctt atgtccactg ggttcgtgcc cgaattgatc acaggcagca
     2521 acgctctgtc atcgttacaa tcaacatgct accctccgcg agatcatccg tgtttcaaac
     2581 ccggcagctt agttgccgtt cttccgaata gcatcggtaa catgagcaaa gtctgccgcc
     2641 ttacaacggc tctcccgctg acgccgtccc ggactgatgg gctgcctgta tcgagtggtg
     2701 attttgtgcc gagctgccgg tcggggagct gttggctggc tggtggcagg atatattgtg
     2761 gtgtaaacaa attgacgctt agacaactta ataacacatt gcggacgttt ttaatgtact
     2821 gaattaacgc cgaattgctc tagccaatac gcaaaccgcc tctccccgcg cgttggccga
     2881 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     2941 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     3001 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgaca
     3061 tgattacgaa ttcaaaaatt acggatatga atataggcat atccgtatcc gaattatccg
     3121 tttgacagct agcaacgatt gtacaattgc ttctttaaaa aaggaagaaa gaaagaaaga
     3181 aaagaatcaa catcagcgtt aacaaacggc cccgttacgg cccaaacggt catatagagt
     3241 aacggcgtta agcgttgaaa gactcctatc gaaatacgta accgcaaacg tgtcatagtc
     3301 agatcccctc ttccttcacc gcctcaaaca caaaaataat cttctacagc ctatatatac
     3361 aaccccccct tctatctctc ctttctcaca attcatcatc tttctttctc tacccccaat
     3421 tttaagaaat cctctcttct cctcttcatt ttcaaggtaa atctctctct ctctctctct
     3481 ctctgttatt ccttgtttta attaggtatg tattattgct agtttgttaa tctgcttatc
     3541 ttatgtatgc cttatgtgaa tatctttatc ttgttcatct catccgttta gaagctataa
     3601 atttgttgat ttgactgtgt atctacacgt ggttatgttt atatctaatc agatatgaat
     3661 ttcttcatat tgttgcgttt gtgtgtacca atccgaaatc gttgattttt ttcatttaat
     3721 cgtgtagcta attgtacgta tacatatgga tctacgtatc aattgttcat ctgtttgtgt
     3781 ttgtatgtat acagatctga aaacatcact tctctcatct gattgtgttg ttacatacat
     3841 agatatagat ctgttatatc atttttttta ttaattgtgt atatatatat gtgcatagat
     3901 ctggattaca tgattgtgat tatttacatg attttgttat ttacgtatgt atatatgtag
     3961 atctggactt tttggagttg ttgacttgat tgtatttgtg tgtgtatatg tgtgttctga
     4021 tcttgatatg ttatgtatgt gcagcgaatt cggcgcgcca tggataagaa gtactctatc
     4081 ggactcgata tcggaactaa ctctgtggga tgggctgtga tcaccgatga gtacaaggtg
     4141 ccatctaaga agttcaaggt tctcggaaac accgataggc actctatcaa gaaaaacctt
     4201 atcggtgctc tcctcttcga ttctggtgaa actgctgagg ctaccagact caagagaacc
     4261 gctagaagaa ggtacaccag aagaaagaac aggatctgct acctccaaga gatcttctct
     4321 aacgagatgg ctaaagtgga tgattcattc ttccacaggc tcgaagagtc attcctcgtg
     4381 gaagaagata agaagcacga gaggcaccct atcttcggaa acatcgttga tgaggtggca
     4441 taccacgaga agtaccctac tatctaccac ctcagaaaga agctcgttga ttctactgat
     4501 aaggctgatc tcaggctcat ctacctcgct ctcgctcaca tgatcaagtt cagaggacac
     4561 ttcctcatcg agggtgatct caaccctgat aactctgatg tggataagtt gttcatccag
     4621 ctcgtgcaga cctacaacca gcttttcgaa gagaacccta tcaacgcttc aggtgtggat
     4681 gctaaggcta tcctctctgc taggctctct aagtcaagaa ggcttgagaa cctcattgct
     4741 cagctccctg gtgagaagaa gaacggactt ttcggaaact tgatcgctct ctctctcgga
     4801 ctcaccccta acttcaagtc taacttcgat ctcgctgagg atgcaaagct ccagctctca
     4861 aaggatacct acgatgatga tctcgataac ctcctcgctc agatcggaga tcagtacgct
     4921 gatttgttcc tcgctgctaa gaacctctct gatgctatcc tcctcagtga tatcctcaga
     4981 gtgaacaccg agatcaccaa ggctccactc tcagcttcta tgatcaagag atacgatgag
     5041 caccaccagg atctcacact tctcaaggct cttgttagac agcagctccc agagaagtac
     5101 aaagagattt tcttcgatca gtctaagaac ggatacgctg gttacatcga tggtggtgca
     5161 tctcaagaag agttctacaa gttcatcaag cctatcctcg agaagatgga tggaaccgag
     5221 gaactcctcg tgaagctcaa tagagaggat cttctcagaa agcagaggac cttcgataac
     5281 ggatctatcc ctcatcagat ccacctcgga gagttgcacg ctatccttag aaggcaagag
     5341 gatttctacc cattcctcaa ggataacagg gaaaagattg agaagattct caccttcaga
     5401 atcccttact acgtgggacc tctcgctaga ggaaactcaa gattcgcttg gatgaccaga
     5461 aagtctgagg aaaccatcac cccttggaac ttcgaagagg tggtggataa gggtgctagt
     5521 gctcagtctt tcatcgagag gatgaccaac ttcgataaga accttccaaa cgagaaggtg
     5581 ctccctaagc actctttgct ctacgagtac ttcaccgtgt acaacgagtt gaccaaggtt
     5641 aagtacgtga ccgagggaat gaggaagcct gcttttttgt caggtgagca aaagaaggct
     5701 atcgttgatc tcttgttcaa gaccaacaga aaggtgaccg tgaagcagct caaagaggat
     5761 tacttcaaga aaatcgagtg cttcgattca gttgagattt ctggtgttga ggataggttc
     5821 aacgcatctc tcggaaccta ccacgatctc ctcaagatca ttaaggataa ggatttcttg
     5881 gataacgagg aaaacgagga tatcttggag gatatcgttc ttaccctcac cctctttgaa
     5941 gatagagaga tgattgaaga aaggctcaag acctacgctc atctcttcga tgataaggtg
     6001 atgaagcagt tgaagagaag aagatacact ggttggggaa ggctctcaag aaagctcatt
     6061 aacggaatca gggataagca gtctggaaag acaatccttg atttcctcaa gtctgatgga
     6121 ttcgctaaca gaaacttcat gcagctcatc cacgatgatt ctctcacctt taaagaggat
     6181 atccagaagg ctcaggtttc aggacagggt gatagtctcc atgagcatat cgctaacctc
     6241 gctggatctc ctgcaatcaa gaagggaatc ctccagactg tgaaggttgt ggatgagttg
     6301 gtgaaggtga tgggaaggca taagcctgag aacatcgtga tcgaaatggc tagagagaac
     6361 cagaccactc agaagggaca gaagaactct agggaaagga tgaagaggat cgaggaaggt
     6421 atcaaagagc ttggatctca gatcctcaaa gagcaccctg ttgagaacac tcagctccag
     6481 aatgagaagc tctacctcta ctacctccag aacggaaggg atatgtatgt ggatcaagag
     6541 ttggatatca acaggctctc tgattacgat gttgatcata tcgtgccaca gtcattcttg
     6601 aaggatgatt ctatcgataa caaggtgctc accaggtctg ataagaacag gggtaagagt
     6661 gataacgtgc caagtgaaga ggttgtgaag aaaatgaaga actattggag gcagctcctc
     6721 aacgctaagc tcatcactca gagaaagttc gataacttga ctaaggctga gaggggagga
     6781 ctctctgaat tggataaggc aggattcatc aagaggcagc ttgtggaaac caggcagatc
     6841 actaagcacg ttgcacagat cctcgattct aggatgaaca ccaagtacga tgagaacgat
     6901 aagttgatca gggaagtgaa ggttatcacc ctcaagtcaa agctcgtgtc tgatttcaga
     6961 aaggatttcc aattctacaa ggtgagggaa atcaacaact accaccacgc tcacgatgct
     7021 taccttaacg ctgttgttgg aaccgctctc atcaagaagt atcctaagct cgagtcagag
     7081 ttcgtgtacg gtgattacaa ggtgtacgat gtgaggaaga tgatcgctaa gtctgagcaa
     7141 gagatcggaa aggctaccgc taagtatttc ttctactcta acatcatgaa tttcttcaag
     7201 accgagatta ccctcgctaa cggtgagatc agaaagaggc cactcatcga gacaaacggt
     7261 gaaacaggtg agatcgtgtg ggataaggga agggatttcg ctaccgttag aaaggtgctc
     7321 tctatgccac aggtgaacat cgttaagaaa accgaggtgc agaccggtgg attctctaaa
     7381 gagtctatcc tccctaagag gaactctgat aagctcattg ctaggaagaa ggattgggac
     7441 cctaagaaat acggtggttt cgattctcct accgtggctt actctgttct cgttgtggct
     7501 aaggttgaga agggaaagag taagaagctc aagtctgtta aggaacttct cggaatcact
     7561 atcatggaaa ggtcatcttt cgagaagaac ccaatcgatt tcctcgaggc taagggatac
     7621 aaagaggtta agaaggatct catcatcaag ctcccaaagt actcactctt cgaactcgag
     7681 aacggtagaa agaggatgct cgcttctgct ggtgagcttc aaaagggaaa cgagcttgct
     7741 ctcccatcta agtacgttaa ctttctttac ctcgcttctc actacgagaa gttgaaggga
     7801 tctccagaag ataacgagca gaagcaactt ttcgttgagc agcacaagca ctacttggat
     7861 gagatcatcg agcagatctc tgagttctct aaaagggtga tcctcgctga tgcaaacctc
     7921 gataaggtgt tgtctgctta caacaagcac agagataagc ctatcaggga acaggcagag
     7981 aacatcatcc atctcttcac ccttaccaac ctcggtgctc ctgctgcttt caagtacttc
     8041 gatacaacca tcgataggaa gagatacacc tctaccaaag aagtgctcga tgctaccctc
     8101 atccatcagt ctatcactgg actctacgag actaggatcg atctctcaca gctcggtggt
     8161 gattcaaggg ctgatcctaa gaagaagagg aaggtttgag gcgcgccgag ctccaggcct
     8221 cccagctttc gtccgtatca tcggtttcga caacgttcgt caagttcaat gcatcagttt
     8281 cattgcccac acaccagaat cctactaagt ttgagtatta tggcattgga aaagctgttt
     8341 tcttctatca tttgttctgc ttgtaattta ctgtgttctt tcagtttttg ttttcggaca
     8401 tcaaaatgca aatggatgga taagagttaa taaatgatat ggtccttttg ttcattctca
     8461 aattattatt atctgttgtt tttactttaa tgggttgaat ttaagtaaga aaggaactaa
     8521 cagtgtgata ttaaggtgca atgttagaca tataaaacag tctttcacct ctctttggtt
     8581 atgtcttgaa ttggtttgtt tcttcactta tctgtgtaat caagtttact atgagtctat
     8641 gatcaagtaa ttatgcaatc aagttaagta cagtataggc ttgagctccc taggcctgtt
     8701 atccctaaca agtttgtaca aaaaagcagg ctggatccac tagtgagctc ctttttttct
     8761 tcttcttcgt tcatacagtt tttttttgtt tatcagctta cattttcttg aaccgtagct
     8821 ttcgttttct tctttttaac tttccattcg gagtttttgt atcttgtttc atagtttgtc
     8881 ccaggattag aatgattagg catcgaacct tcaagaattt gattgaataa aacatcttca
     8941 ttcttaagat atgaagataa tcttcaaaag gcccctggga atctgaaaga agagaagcag
     9001 gcccatttat atgggaaaga acaatagtat ttcttatata ggcccattta agttgaaaac
     9061 aatcttcaaa agtcccacat cgcttagata agaaaacgaa gctgagttta tatacagcta
     9121 gagtcgaagt agtgattgat ggagaatctg taaagctcgt ttaagagcta tgctggaaac
     9181 agcatagcaa gtttaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg
     9241 gtgctttttt tgttttttat gtctagctct agctaacgcc ggcggttcag ttgcattatg
     9301 tctttataca cccctaccta gatgattaag ttttacttta gttggtgtga aatggataaa
     9361 ttctaaaata tgaggtgtgg aatgaaggat tgtcatcaat tagttggccc caaccaagta
     9421 aaataagaag gccggcccat tacaattaag tcgtcacaca agtgggcttc attgaaacaa
     9481 gcgcaaaaag gagtccaggc ccgtgttagc gtgaagactc aaccagcgat ttctccctca
     9541 tcggttgcac agaaaagctg tgtgttgttt atatggcgaa acctaacagt ctgacttgca
     9601 gatcaattga atcatccagt ttaagagcta tgctggaaac agcatagcaa gtttaaataa
     9661 ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt tgttttttat
     9721 gtctggtgac cataggacat cggccgagct gaatagtacg cgttacgact tatcggcgcg
     9781 ccgaattaga tcacctgagg gagctcacta gtggatccac ccagctttct tgtacaaagt
     9841 ggttcgataa ttccgatcca gcctaggccc gggcctgagg acgcgtccat ggttaattaa
     9901 gacgtccgga ccgactagtg gatcctctag agtcgacctg caggcatgca agcttcagct
     9961 tgcatgccgg tcgatctagt aacatagatg acaccgcgcg cgataattta tcctagtttg
    10021 cgcgctatat tttgttttct atcgcgtatt aaatgtataa ttgcgggact ctaatcataa
    10081 aaacccatct cataaataac gtcatgcatt acatgttaat tattacatgc ttaacgtaat
    10141 tcaacagaaa ttatatgata atcatcgcaa gaccggcaac aggattcaat cttaagaagc
    10201 tttattgcca aatgtttgaa cgatctgctt gactctagct agagtccgaa ccccagagtc
    10261 ccgctcagaa gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg
    10321 cgataccgta aagcacgagg aagcggtcag cccattcgcc gccaagctct tcagcaatat
    10381 cacgggtagc caacgctatg tcctgatagc ggtccgccac acccagccgg ccacagtcga
    10441 tgaatccaga aaagcggcca ttttccacca tgatattcgg caagcaggca tcgccgtggg
    10501 tcacgacgag atcctcgccg tcgggcatcc gcgccttgag cctggcgaac agttcggctg
    10561 gcgcgagccc ctgatgctct tcgtccagat catcctgatc gacaagaccg gcttccatcc
    10621 gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag gtagccggat
    10681 caagcgtatg cagccgccgc attgcatcag ccatgatgga tactttctcg gcaggagcaa
    10741 ggtgagatga caggagatcc tgccccggca cttcgcccaa tagcagccag tcccttcccg
    10801 cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc agccacgata
    10861 gccgcgctgc ctcgtcttgg agttcattca gggcaccgga caggtcggtc ttgacaaaaa
    10921 gaaccgggcg cccctgcgct gacagccgga acacggcggc atcagagcag ccgattgtct
    10981 gttgtgccca gtcatagccg aatagcctct ccacccaagc ggccggagaa cctgcgtgca
    11041 atccatcttg ttcaatcatg cctcgatcga gttgagggtg aatatgagac tctaattgga
    11101 taccgagggg aatttatgga acgtcagtgg agcatttttg acaagaaata tttgctagct
    11161 gatagtgacc ttaggcgact tttgaacgcg caataatggt ttctgacgta tgtgcttagc
    11221 tcattaaact ccagaaaccc gcggctcagt ggctccttca acgttgcggt tctgtcagtt
    11281 ccaaacgtaa aacggcttgt cccgcgtcat cggcgggggt cataacgtga ctcccttaat
    11341 tctcatgtat gaaagcttgg cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
    11401 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
    11461 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatgagc
    11521 ttgagcttgg atcagattgt cgtttcccgc cttcagttta aactatcagt gtttgacagg
    11581 atatattggc gggtaaacct aagagaaaag agcgtttatt agaataatcg gatatttaaa
    11641 agggcgtgaa aaggtttatc cgttcgtcca tttgtatgtg catgccaacc acagggttcc
    11701 cctcgggatc aaagtacttt aaagtacttt aaagtacttt aaagtacttt gatccaaccc
    11761 ctccgctgct atagtgcagt cggcttctga cgttcagtgc agccgtcttc tgaaaacgac
    11821 atgtcgcaca agtcctaagt tacgcgacag gctgccgccc tgcccttttc ctggcgtttt
    11881 cttgtcgcgt gttttagtcg cataaagtag aatacttgcg actagaaccg gagacattac
    11941 gccatgaaca agagcgccgc cgctggcctg ctgggctatg cccgcgtcag caccgacgac
    12001 caggacttga ccaaccaacg ggccgaactg cacgcggccg gctgcaccaa gctgttttcc
    12061 gagaagatca ccggcaccag gcgcgaccgc ccggagctgg ccaggatgct tgaccaccta
    12121 cgccctggcg acgttgtgac agtgaccagg ctagaccgcc tggcccgcag cacccgcgac
    12181 ctactggaca ttgccgagcg catccaggag gccggcgcgg gcctgcgtag cctggcagag
    12241 ccgtgggccg acaccaccac gccggccggc cgcatggtgt tgaccgtgtt cgccggcatt
    12301 gccgagttcg agcgttccct aatcatcgac cgcacccgga gcgggcgcga ggccgccaag
    12361 gcccgaggcg tgaagtttgg cccccgccct accctcaccc cggcacagat cgcgcacgcc
    12421 cgcgagctga tcgaccagga aggccgcacc gtgaaagagg cggctgcact gcttggcgtg
    12481 catcgctcga ccctgtaccg cgcacttgag cgcagcgagg aagtgacgcc caccgaggcc
    12541 aggcggcgcg gtgccttccg tgaggacgca ttgaccgagg ccgacgccct ggcggccgcc
    12601 gagaatgaac gccaagagga acaagcatga aaccgcacca ggacggccag gacgaaccgt
    12661 ttttcattac cgaagagatc gaggcggaga tgatcgcggc cgggtacgtg ttcgagccgc
    12721 ccgcgcacgt ctcaaccgtg cggctgcatg aaatcctggc cggtttgtct gatgccaagc
    12781 tggcggcctg gccggccagc ttggccgctg aagaaaccga gcgccgccgt ctaaaaaggt
    12841 gatgtgtatt tgagtaaaac agcttgcgtc atgcggtcgc tgcgtatatg atgcgatgag
    12901 taaataaaca aatacgcaag gggaacgcat gaaggttatc gctgtactta accagaaagg
    12961 cgggtcaggc aagacgacca tcgcaaccca tctagcccgc gccctgcaac tcgccggggc
    13021 cgatgttctg ttagtcgatt ccgatcccca gggcagtgcc cgcgattggg cggccgtgcg
    13081 ggaagatcaa ccgctaaccg ttgtcggcat cgaccgcccg acgattgacc gcgacgtgaa
    13141 ggccatcggc cggcgcgact tcgtagtgat cgacggagcg ccccaggcgg cggacttggc
    13201 tgtgtccgcg atcaaggcag ccgacttcgt gctgattccg gtgcagccaa gcccttacga
    13261 catatgggcc accgccgacc tggtggagct ggttaagcag cgcattgagg tcacggatgg
    13321 aaggctacaa gcggcctttg tcgtgtcgcg ggcgatcaaa ggcacgcgca tcggcggtga
    13381 ggttgccgag gcgctggccg ggtacgagct gcccattctt gagtcccgta tcacgcagcg
    13441 cgtgagctac ccaggcactg ccgccgccgg cacaaccgtt cttgaatcag aacccgaggg
    13501 cgacgctgcc cgcgaggtcc aggcgctggc cgctgaaatt aaatcaaaac tcatttgagt
    13561 taatgaggta aagagaaaat gagcaaaagc acaaacacgc taagtgccgg ccgtccgagc
    13621 gcacgcagca gcaaggctgc aacgttggcc agcctggcag acacgccagc catgaagcgg
    13681 gtcaactttc agttgccggc ggaggatcac accaagctga agatgtacgc ggtacgccaa
    13741 ggcaagacca ttaccgagct gctatctgaa tacatcgcgc agctaccaga gtaaatgagc
    13801 aaatgaataa atgagtagat gaattttagc ggctaaagga ggcggcatgg aaaatcaaga
    13861 acaaccaggc accgacgccg tggaatgccc catgtgtgga ggaacgggcg gttggccagg
    13921 cgtaagcggc tgggttgtct gccggccctg caatggcact ggaaccccca agcccgagga
    13981 atcggcgtga gcggtcgcaa accatccggc ccggtacaaa tcggcgcggc gctgggtgat
    14041 gacctggtgg agaagttgaa ggccgcgcag gccgcccagc ggcaacgcat cgaggcagaa
    14101 gcacgccccg gtgaatcgtg gcaagcggcc gctgatcgaa tccgcaaaga atcccggcaa
    14161 ccgccggcag ccggtgcgcc gtcgattagg aagccgccca agggcgacga gcaaccagat
    14221 tttttcgttc cgatgctcta tgacgtgggc acccgcgata gtcgcagcat catggacgtg
    14281 gccgttttcc gtctgtcgaa gcgtgaccga cgagctggcg aggtgatccg ctacgagctt
    14341 ccagacgggc acgtagaggt ttccgcaggg ccggccggca tggccagtgt gtgggattac
    14401 gacctggtac tgatggcggt ttcccatcta accgaatcca tgaaccgata ccgggaaggg
    14461 aagggagaca agcccggccg cgtgttccgt ccacacgttg cggacgtact caagttctgc
    14521 cggcgagccg atggcggaaa gcagaaagac gacctggtag aaacctgcat tcggttaaac
    14581 accacgcacg ttgccatgca gcgtacgaag aaggccaaga acggccgcct ggtgacggta
    14641 tccgagggtg aagccttgat tagccgctac aagatcgtaa agagcgaaac cgggcggccg
    14701 gagtacatcg agatcgagct agctgattgg atgtaccgcg agatcacaga aggcaagaac
    14761 ccggacgtgc tgacggttca ccccgattac tttttgatcg atcccggcat cggccgtttt
    14821 ctctaccgcc tggcacgccg cgccgcaggc aaggcagaag ccagatggtt gttcaagacg
    14881 atctacgaac gcagtggcag cgccggagag ttcaagaagt tctgtttcac cgtgcgcaag
    14941 ctgatcgggt caaatgacct gccggagtac gatttgaagg aggaggcggg gcaggctggc
    15001 ccgatcctag tcatgcgcta ccgcaacctg atcgagggcg aagcatccgc cggttcctaa
    15061 tgtacggagc agatgctagg gcaaattgcc ctagcagggg aaaaaggtcg aaaaggtctc
    15121 tttcctgtgg atagcacgta cattgggaac ccaaagccgt acattgggaa ccggaacccg
    15181 tacattggga acccaaagcc gtacattggg aaccggtcac acatgtaagt gactgatata
    15241 aaagagaaaa aaggcgattt ttccgcctaa aactctttaa aacttattaa aactcttaaa
    15301 acccgcctgg cctgtgcata actgtctggc cagcgcacag ccgaagagct gcaaaaagcg
    15361 cctacccttc ggtcgctgcg ctccctacgc cc
//
