LOCUS       sequence-351582-       15392 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Binary expression vector with A.th. codon-optimized Cas9 and sgRNA 
            for miRNA160b.
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 Apr 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_R160b
FEATURES             Location/Qualifiers
     source          1..15392
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(98..889)
                     /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    1410..1434
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     protein_bind    1611..1632
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1647..1652,1653..1670,1671..1677)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1685..1701
                     /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     1690..1712
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1709..1725
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1709..1725
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2726..6829
                     /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             2726..6829
                     /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             6842..6862
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    7374..7398
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      7911..7930
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     terminator      8371..8390
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     protein_bind    complement(8485..8509)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      complement(8639..8891)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(8931..9725)
                     /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(9042..9061)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     9652..9671
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(9759..9938)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     complement(10028..10045)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10028..10044)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10037..10059)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    10240..10264
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             11595..12224
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             12653..13726
                     /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      13792..13986
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(14222..14244)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    14330..14470
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     14385..14404
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(14485..14502)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(14656..15244)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(14736..14755)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agtttggctg tgagcaatta tgtgcttagt gcatctaacg cttgagttaa gccgcgccgc
       61 gaagcggcgt cggcttgaac gaatttctag ctagacatta tttgccgact accttggtga
      121 tctcgccttt cacgtagtgg acaaattctt ccaactgatc tgcgcgcgag gccaagcgat
      181 cttcttcttg tccaagataa gcctgtctag cttcaagtat gacgggctga tactgggccg
      241 gcaggcgctc cattgcccag tcggcagcga catccttcgg cgcgattttg ccggttactg
      301 cgctgtacca aatgcgggac aacgtaagca ctacatttcg ctcatcgcca gcccagtcgg
      361 gcggcgagtt ccatagcgtt aaggtttcat ttagcgcctc aaatagatcc tgttcaggaa
      421 ccggatcaaa gagttcctcc gccgctggac ctaccaaggc aacgctatgt tctcttgctt
      481 ttgtcagcaa gatagccaga tcaatgtcga tcgtggctgg ctcgaagata cctgcaagaa
      541 tgtcattgcg ctgccattct ccaaattgca gttcgcgctt agctggataa cgccacggaa
      601 tgatgtcgtc gtgcacaaca atggtgactt ctacagcgcg gagaatctcg ctctctccag
      661 gggaagccga agtttccaaa aggtcgttga tcaaagctcg ccgcgttgtt tcatcaagcc
      721 ttacggtcac cgtaaccagc aaatcaatat cactgtgtgg cttcaggccg ccatccactg
      781 cggagccgta caaatgtacg gccagcaacg tcggttcgag atggcgctcg atgacgccaa
      841 ctacctctga tagttgagtc gatacttcgg cgatcaccgc ttcccccatg atgtttaact
      901 ttgttttagg gcgactgccc tgctgcgtaa catcgttgct gctccataac atcaaacatc
      961 gacccacggc gtaacgcgct tgctgcttgg atgcccgagg catagactgt accccaaaaa
     1021 aacagtcata acaagccatg aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca
     1081 aggttctgga ccagttgcgt gagcgcatac gctacttgca ttacagctta cgaaccgaac
     1141 aggcttatgt ccactgggtt cgtgcccgaa ttgatcacag gcagcaacgc tctgtcatcg
     1201 ttacaatcaa catgctaccc tccgcgagat catccgtgtt tcaaacccgg cagcttagtt
     1261 gccgttcttc cgaatagcat cggtaacatg agcaaagtct gccgccttac aacggctctc
     1321 ccgctgacgc cgtcccggac tgatgggctg cctgtatcga gtggtgattt tgtgccgagc
     1381 tgccggtcgg ggagctgttg gctggctggt ggcaggatat attgtggtgt aaacaaattg
     1441 acgcttagac aacttaataa cacattgcgg acgtttttaa tgtactgaat taacgccgaa
     1501 ttgctctagc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     1561 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     1621 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     1681 ggaattgtga gcggataaca atttcacaca ggaaacagct atgacatgat tacgaattca
     1741 aaaattacgg atatgaatat aggcatatcc gtatccgaat tatccgtttg acagctagca
     1801 acgattgtac aattgcttct ttaaaaaagg aagaaagaaa gaaagaaaag aatcaacatc
     1861 agcgttaaca aacggccccg ttacggccca aacggtcata tagagtaacg gcgttaagcg
     1921 ttgaaagact cctatcgaaa tacgtaaccg caaacgtgtc atagtcagat cccctcttcc
     1981 ttcaccgcct caaacacaaa aataatcttc tacagcctat atatacaacc cccccttcta
     2041 tctctccttt ctcacaattc atcatctttc tttctctacc cccaatttta agaaatcctc
     2101 tcttctcctc ttcattttca aggtaaatct ctctctctct ctctctctct gttattcctt
     2161 gttttaatta ggtatgtatt attgctagtt tgttaatctg cttatcttat gtatgcctta
     2221 tgtgaatatc tttatcttgt tcatctcatc cgtttagaag ctataaattt gttgatttga
     2281 ctgtgtatct acacgtggtt atgtttatat ctaatcagat atgaatttct tcatattgtt
     2341 gcgtttgtgt gtaccaatcc gaaatcgttg atttttttca tttaatcgtg tagctaattg
     2401 tacgtataca tatggatcta cgtatcaatt gttcatctgt ttgtgtttgt atgtatacag
     2461 atctgaaaac atcacttctc tcatctgatt gtgttgttac atacatagat atagatctgt
     2521 tatatcattt tttttattaa ttgtgtatat atatatgtgc atagatctgg attacatgat
     2581 tgtgattatt tacatgattt tgttatttac gtatgtatat atgtagatct ggactttttg
     2641 gagttgttga cttgattgta tttgtgtgtg tatatgtgtg ttctgatctt gatatgttat
     2701 gtatgtgcag cgaattcggc gcgccatgga taagaagtac tctatcggac tcgatatcgg
     2761 aactaactct gtgggatggg ctgtgatcac cgatgagtac aaggtgccat ctaagaagtt
     2821 caaggttctc ggaaacaccg ataggcactc tatcaagaaa aaccttatcg gtgctctcct
     2881 cttcgattct ggtgaaactg ctgaggctac cagactcaag agaaccgcta gaagaaggta
     2941 caccagaaga aagaacagga tctgctacct ccaagagatc ttctctaacg agatggctaa
     3001 agtggatgat tcattcttcc acaggctcga agagtcattc ctcgtggaag aagataagaa
     3061 gcacgagagg caccctatct tcggaaacat cgttgatgag gtggcatacc acgagaagta
     3121 ccctactatc taccacctca gaaagaagct cgttgattct actgataagg ctgatctcag
     3181 gctcatctac ctcgctctcg ctcacatgat caagttcaga ggacacttcc tcatcgaggg
     3241 tgatctcaac cctgataact ctgatgtgga taagttgttc atccagctcg tgcagaccta
     3301 caaccagctt ttcgaagaga accctatcaa cgcttcaggt gtggatgcta aggctatcct
     3361 ctctgctagg ctctctaagt caagaaggct tgagaacctc attgctcagc tccctggtga
     3421 gaagaagaac ggacttttcg gaaacttgat cgctctctct ctcggactca cccctaactt
     3481 caagtctaac ttcgatctcg ctgaggatgc aaagctccag ctctcaaagg atacctacga
     3541 tgatgatctc gataacctcc tcgctcagat cggagatcag tacgctgatt tgttcctcgc
     3601 tgctaagaac ctctctgatg ctatcctcct cagtgatatc ctcagagtga acaccgagat
     3661 caccaaggct ccactctcag cttctatgat caagagatac gatgagcacc accaggatct
     3721 cacacttctc aaggctcttg ttagacagca gctcccagag aagtacaaag agattttctt
     3781 cgatcagtct aagaacggat acgctggtta catcgatggt ggtgcatctc aagaagagtt
     3841 ctacaagttc atcaagccta tcctcgagaa gatggatgga accgaggaac tcctcgtgaa
     3901 gctcaataga gaggatcttc tcagaaagca gaggaccttc gataacggat ctatccctca
     3961 tcagatccac ctcggagagt tgcacgctat ccttagaagg caagaggatt tctacccatt
     4021 cctcaaggat aacagggaaa agattgagaa gattctcacc ttcagaatcc cttactacgt
     4081 gggacctctc gctagaggaa actcaagatt cgcttggatg accagaaagt ctgaggaaac
     4141 catcacccct tggaacttcg aagaggtggt ggataagggt gctagtgctc agtctttcat
     4201 cgagaggatg accaacttcg ataagaacct tccaaacgag aaggtgctcc ctaagcactc
     4261 tttgctctac gagtacttca ccgtgtacaa cgagttgacc aaggttaagt acgtgaccga
     4321 gggaatgagg aagcctgctt ttttgtcagg tgagcaaaag aaggctatcg ttgatctctt
     4381 gttcaagacc aacagaaagg tgaccgtgaa gcagctcaaa gaggattact tcaagaaaat
     4441 cgagtgcttc gattcagttg agatttctgg tgttgaggat aggttcaacg catctctcgg
     4501 aacctaccac gatctcctca agatcattaa ggataaggat ttcttggata acgaggaaaa
     4561 cgaggatatc ttggaggata tcgttcttac cctcaccctc tttgaagata gagagatgat
     4621 tgaagaaagg ctcaagacct acgctcatct cttcgatgat aaggtgatga agcagttgaa
     4681 gagaagaaga tacactggtt ggggaaggct ctcaagaaag ctcattaacg gaatcaggga
     4741 taagcagtct ggaaagacaa tccttgattt cctcaagtct gatggattcg ctaacagaaa
     4801 cttcatgcag ctcatccacg atgattctct cacctttaaa gaggatatcc agaaggctca
     4861 ggtttcagga cagggtgata gtctccatga gcatatcgct aacctcgctg gatctcctgc
     4921 aatcaagaag ggaatcctcc agactgtgaa ggttgtggat gagttggtga aggtgatggg
     4981 aaggcataag cctgagaaca tcgtgatcga aatggctaga gagaaccaga ccactcagaa
     5041 gggacagaag aactctaggg aaaggatgaa gaggatcgag gaaggtatca aagagcttgg
     5101 atctcagatc ctcaaagagc accctgttga gaacactcag ctccagaatg agaagctcta
     5161 cctctactac ctccagaacg gaagggatat gtatgtggat caagagttgg atatcaacag
     5221 gctctctgat tacgatgttg atcatatcgt gccacagtca ttcttgaagg atgattctat
     5281 cgataacaag gtgctcacca ggtctgataa gaacaggggt aagagtgata acgtgccaag
     5341 tgaagaggtt gtgaagaaaa tgaagaacta ttggaggcag ctcctcaacg ctaagctcat
     5401 cactcagaga aagttcgata acttgactaa ggctgagagg ggaggactct ctgaattgga
     5461 taaggcagga ttcatcaaga ggcagcttgt ggaaaccagg cagatcacta agcacgttgc
     5521 acagatcctc gattctagga tgaacaccaa gtacgatgag aacgataagt tgatcaggga
     5581 agtgaaggtt atcaccctca agtcaaagct cgtgtctgat ttcagaaagg atttccaatt
     5641 ctacaaggtg agggaaatca acaactacca ccacgctcac gatgcttacc ttaacgctgt
     5701 tgttggaacc gctctcatca agaagtatcc taagctcgag tcagagttcg tgtacggtga
     5761 ttacaaggtg tacgatgtga ggaagatgat cgctaagtct gagcaagaga tcggaaaggc
     5821 taccgctaag tatttcttct actctaacat catgaatttc ttcaagaccg agattaccct
     5881 cgctaacggt gagatcagaa agaggccact catcgagaca aacggtgaaa caggtgagat
     5941 cgtgtgggat aagggaaggg atttcgctac cgttagaaag gtgctctcta tgccacaggt
     6001 gaacatcgtt aagaaaaccg aggtgcagac cggtggattc tctaaagagt ctatcctccc
     6061 taagaggaac tctgataagc tcattgctag gaagaaggat tgggacccta agaaatacgg
     6121 tggtttcgat tctcctaccg tggcttactc tgttctcgtt gtggctaagg ttgagaaggg
     6181 aaagagtaag aagctcaagt ctgttaagga acttctcgga atcactatca tggaaaggtc
     6241 atctttcgag aagaacccaa tcgatttcct cgaggctaag ggatacaaag aggttaagaa
     6301 ggatctcatc atcaagctcc caaagtactc actcttcgaa ctcgagaacg gtagaaagag
     6361 gatgctcgct tctgctggtg agcttcaaaa gggaaacgag cttgctctcc catctaagta
     6421 cgttaacttt ctttacctcg cttctcacta cgagaagttg aagggatctc cagaagataa
     6481 cgagcagaag caacttttcg ttgagcagca caagcactac ttggatgaga tcatcgagca
     6541 gatctctgag ttctctaaaa gggtgatcct cgctgatgca aacctcgata aggtgttgtc
     6601 tgcttacaac aagcacagag ataagcctat cagggaacag gcagagaaca tcatccatct
     6661 cttcaccctt accaacctcg gtgctcctgc tgctttcaag tacttcgata caaccatcga
     6721 taggaagaga tacacctcta ccaaagaagt gctcgatgct accctcatcc atcagtctat
     6781 cactggactc tacgagacta ggatcgatct ctcacagctc ggtggtgatt caagggctga
     6841 tcctaagaag aagaggaagg tttgaggcgc gccgagctcc aggcctccca gctttcgtcc
     6901 gtatcatcgg tttcgacaac gttcgtcaag ttcaatgcat cagtttcatt gcccacacac
     6961 cagaatccta ctaagtttga gtattatggc attggaaaag ctgttttctt ctatcatttg
     7021 ttctgcttgt aatttactgt gttctttcag tttttgtttt cggacatcaa aatgcaaatg
     7081 gatggataag agttaataaa tgatatggtc cttttgttca ttctcaaatt attattatct
     7141 gttgttttta ctttaatggg ttgaatttaa gtaagaaagg aactaacagt gtgatattaa
     7201 ggtgcaatgt tagacatata aaacagtctt tcacctctct ttggttatgt cttgaattgg
     7261 tttgtttctt cacttatctg tgtaatcaag tttactatga gtctatgatc aagtaattat
     7321 gcaatcaagt taagtacagt ataggcttga gctccctagg cctgttatcc ctaacaagtt
     7381 tgtacaaaaa agcaggctgg atccactagt gagctccttt ttttcttctt cttcgttcat
     7441 acagtttttt tttgtttatc agcttacatt ttcttgaacc gtagctttcg ttttcttctt
     7501 tttaactttc cattcggagt ttttgtatct tgtttcatag tttgtcccag gattagaatg
     7561 attaggcatc gaaccttcaa gaatttgatt gaataaaaca tcttcattct taagatatga
     7621 agataatctt caaaaggccc ctgggaatct gaaagaagag aagcaggccc atttatatgg
     7681 gaaagaacaa tagtatttct tatataggcc catttaagtt gaaaacaatc ttcaaaagtc
     7741 ccacatcgct tagataagaa aacgaagctg agtttatata cagctagagt cgaagtagtg
     7801 attgaggagt aagaatgatg tgccgtttaa gagctatgct ggaaacagca tagcaagttt
     7861 aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc tttttttgtt
     7921 ttttatgtct agctctagct aacgccggcg gttcagttgc attatgtctt tatacacccc
     7981 tacctagatg attaagtttt actttagttg gtgtgaaatg gataaattct aaaatatgag
     8041 gtgtggaatg aaggattgtc atcaattagt tggccccaac caagtaaaat aagaaggccg
     8101 gcccattaca attaagtcgt cacacaagtg ggcttcattg aaacaagcgc aaaaaggagt
     8161 ccaggcccgt gttagcgtga agactcaacc agcgatttct ccctcatcgg ttgcacagaa
     8221 aagctgtgtg ttgtttatat ggcgaaacct aacagtctga cttgggtgaa agtgtgtggc
     8281 atacgtttaa gagctatgct ggaaacagca tagcaagttt aaataaggct agtccgttat
     8341 caacttgaaa aagtggcacc gagtcggtgc tttttttgtt ttttatgtct ggtgaccata
     8401 ggacatcggc cgagctgaat agtacgcgtt acgacttatc ggcgcgccga attagatcac
     8461 ctgagggagc tcactagtgg atccacccag ctttcttgta caaagtggtt cgataattcc
     8521 gatccagcct aggcccgggc ctgaggacgc gtccatggtt aattaagacg tccggaccga
     8581 ctagtggatc ctctagagtc gacctgcagg catgcaagct tcagcttgca tgccggtcga
     8641 tctagtaaca tagatgacac cgcgcgcgat aatttatcct agtttgcgcg ctatattttg
     8701 ttttctatcg cgtattaaat gtataattgc gggactctaa tcataaaaac ccatctcata
     8761 aataacgtca tgcattacat gttaattatt acatgcttaa cgtaattcaa cagaaattat
     8821 atgataatca tcgcaagacc ggcaacagga ttcaatctta agaagcttta ttgccaaatg
     8881 tttgaacgat ctgcttgact ctagctagag tccgaacccc agagtcccgc tcagaagaac
     8941 tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc
     9001 acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac
     9061 gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag
     9121 cggccatttt ccaccatgat attcggcaag caggcatcgc cgtgggtcac gacgagatcc
     9181 tcgccgtcgg gcatccgcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga
     9241 tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc
     9301 tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc
     9361 cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg
     9421 agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg
     9481 tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg
     9541 tcttggagtt cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc
     9601 tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca
     9661 tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca
     9721 atcatgcctc gatcgagttg agggtgaata tgagactcta attggatacc gaggggaatt
     9781 tatggaacgt cagtggagca tttttgacaa gaaatatttg ctagctgata gtgaccttag
     9841 gcgacttttg aacgcgcaat aatggtttct gacgtatgtg cttagctcat taaactccag
     9901 aaacccgcgg ctcagtggct ccttcaacgt tgcggttctg tcagttccaa acgtaaaacg
     9961 gcttgtcccg cgtcatcggc gggggtcata acgtgactcc cttaattctc atgtatgaaa
    10021 gcttggcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
    10081 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
    10141 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgagcttga gcttggatca
    10201 gattgtcgtt tcccgccttc agtttaaact atcagtgttt gacaggatat attggcgggt
    10261 aaacctaaga gaaaagagcg tttattagaa taatcggata tttaaaaggg cgtgaaaagg
    10321 tttatccgtt cgtccatttg tatgtgcatg ccaaccacag ggttcccctc gggatcaaag
    10381 tactttaaag tactttaaag tactttaaag tactttgatc caacccctcc gctgctatag
    10441 tgcagtcggc ttctgacgtt cagtgcagcc gtcttctgaa aacgacatgt cgcacaagtc
    10501 ctaagttacg cgacaggctg ccgccctgcc cttttcctgg cgttttcttg tcgcgtgttt
    10561 tagtcgcata aagtagaata cttgcgacta gaaccggaga cattacgcca tgaacaagag
    10621 cgccgccgct ggcctgctgg gctatgcccg cgtcagcacc gacgaccagg acttgaccaa
    10681 ccaacgggcc gaactgcacg cggccggctg caccaagctg ttttccgaga agatcaccgg
    10741 caccaggcgc gaccgcccgg agctggccag gatgcttgac cacctacgcc ctggcgacgt
    10801 tgtgacagtg accaggctag accgcctggc ccgcagcacc cgcgacctac tggacattgc
    10861 cgagcgcatc caggaggccg gcgcgggcct gcgtagcctg gcagagccgt gggccgacac
    10921 caccacgccg gccggccgca tggtgttgac cgtgttcgcc ggcattgccg agttcgagcg
    10981 ttccctaatc atcgaccgca cccggagcgg gcgcgaggcc gccaaggccc gaggcgtgaa
    11041 gtttggcccc cgccctaccc tcaccccggc acagatcgcg cacgcccgcg agctgatcga
    11101 ccaggaaggc cgcaccgtga aagaggcggc tgcactgctt ggcgtgcatc gctcgaccct
    11161 gtaccgcgca cttgagcgca gcgaggaagt gacgcccacc gaggccaggc ggcgcggtgc
    11221 cttccgtgag gacgcattga ccgaggccga cgccctggcg gccgccgaga atgaacgcca
    11281 agaggaacaa gcatgaaacc gcaccaggac ggccaggacg aaccgttttt cattaccgaa
    11341 gagatcgagg cggagatgat cgcggccggg tacgtgttcg agccgcccgc gcacgtctca
    11401 accgtgcggc tgcatgaaat cctggccggt ttgtctgatg ccaagctggc ggcctggccg
    11461 gccagcttgg ccgctgaaga aaccgagcgc cgccgtctaa aaaggtgatg tgtatttgag
    11521 taaaacagct tgcgtcatgc ggtcgctgcg tatatgatgc gatgagtaaa taaacaaata
    11581 cgcaagggga acgcatgaag gttatcgctg tacttaacca gaaaggcggg tcaggcaaga
    11641 cgaccatcgc aacccatcta gcccgcgccc tgcaactcgc cggggccgat gttctgttag
    11701 tcgattccga tccccagggc agtgcccgcg attgggcggc cgtgcgggaa gatcaaccgc
    11761 taaccgttgt cggcatcgac cgcccgacga ttgaccgcga cgtgaaggcc atcggccggc
    11821 gcgacttcgt agtgatcgac ggagcgcccc aggcggcgga cttggctgtg tccgcgatca
    11881 aggcagccga cttcgtgctg attccggtgc agccaagccc ttacgacata tgggccaccg
    11941 ccgacctggt ggagctggtt aagcagcgca ttgaggtcac ggatggaagg ctacaagcgg
    12001 cctttgtcgt gtcgcgggcg atcaaaggca cgcgcatcgg cggtgaggtt gccgaggcgc
    12061 tggccgggta cgagctgccc attcttgagt cccgtatcac gcagcgcgtg agctacccag
    12121 gcactgccgc cgccggcaca accgttcttg aatcagaacc cgagggcgac gctgcccgcg
    12181 aggtccaggc gctggccgct gaaattaaat caaaactcat ttgagttaat gaggtaaaga
    12241 gaaaatgagc aaaagcacaa acacgctaag tgccggccgt ccgagcgcac gcagcagcaa
    12301 ggctgcaacg ttggccagcc tggcagacac gccagccatg aagcgggtca actttcagtt
    12361 gccggcggag gatcacacca agctgaagat gtacgcggta cgccaaggca agaccattac
    12421 cgagctgcta tctgaataca tcgcgcagct accagagtaa atgagcaaat gaataaatga
    12481 gtagatgaat tttagcggct aaaggaggcg gcatggaaaa tcaagaacaa ccaggcaccg
    12541 acgccgtgga atgccccatg tgtggaggaa cgggcggttg gccaggcgta agcggctggg
    12601 ttgtctgccg gccctgcaat ggcactggaa cccccaagcc cgaggaatcg gcgtgagcgg
    12661 tcgcaaacca tccggcccgg tacaaatcgg cgcggcgctg ggtgatgacc tggtggagaa
    12721 gttgaaggcc gcgcaggccg cccagcggca acgcatcgag gcagaagcac gccccggtga
    12781 atcgtggcaa gcggccgctg atcgaatccg caaagaatcc cggcaaccgc cggcagccgg
    12841 tgcgccgtcg attaggaagc cgcccaaggg cgacgagcaa ccagattttt tcgttccgat
    12901 gctctatgac gtgggcaccc gcgatagtcg cagcatcatg gacgtggccg ttttccgtct
    12961 gtcgaagcgt gaccgacgag ctggcgaggt gatccgctac gagcttccag acgggcacgt
    13021 agaggtttcc gcagggccgg ccggcatggc cagtgtgtgg gattacgacc tggtactgat
    13081 ggcggtttcc catctaaccg aatccatgaa ccgataccgg gaagggaagg gagacaagcc
    13141 cggccgcgtg ttccgtccac acgttgcgga cgtactcaag ttctgccggc gagccgatgg
    13201 cggaaagcag aaagacgacc tggtagaaac ctgcattcgg ttaaacacca cgcacgttgc
    13261 catgcagcgt acgaagaagg ccaagaacgg ccgcctggtg acggtatccg agggtgaagc
    13321 cttgattagc cgctacaaga tcgtaaagag cgaaaccggg cggccggagt acatcgagat
    13381 cgagctagct gattggatgt accgcgagat cacagaaggc aagaacccgg acgtgctgac
    13441 ggttcacccc gattactttt tgatcgatcc cggcatcggc cgttttctct accgcctggc
    13501 acgccgcgcc gcaggcaagg cagaagccag atggttgttc aagacgatct acgaacgcag
    13561 tggcagcgcc ggagagttca agaagttctg tttcaccgtg cgcaagctga tcgggtcaaa
    13621 tgacctgccg gagtacgatt tgaaggagga ggcggggcag gctggcccga tcctagtcat
    13681 gcgctaccgc aacctgatcg agggcgaagc atccgccggt tcctaatgta cggagcagat
    13741 gctagggcaa attgccctag caggggaaaa aggtcgaaaa ggtctctttc ctgtggatag
    13801 cacgtacatt gggaacccaa agccgtacat tgggaaccgg aacccgtaca ttgggaaccc
    13861 aaagccgtac attgggaacc ggtcacacat gtaagtgact gatataaaag agaaaaaagg
    13921 cgatttttcc gcctaaaact ctttaaaact tattaaaact cttaaaaccc gcctggcctg
    13981 tgcataactg tctggccagc gcacagccga agagctgcaa aaagcgccta cccttcggtc
    14041 gctgcgctcc ctacgccccg ccgcttcgcg tcggcctatc gcggccgctg gccgctcaaa
    14101 aatggctggc ctacggccag gcaatctacc agggcgcgga caagccgcgc cgtcgccact
    14161 cgaccgccgg cgcccacatc aaggcaccct gcctcgcgcg tttcggtgat gacggtgaaa
    14221 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
    14281 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga
    14341 cccagtcacg tagcgatagc ggagtgtata ctggcttaac tatgcggcat cagagcagat
    14401 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
    14461 ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
    14521 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
    14581 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
    14641 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
    14701 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
    14761 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
    14821 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
    14881 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
    14941 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
    15001 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
    15061 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
    15121 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
    15181 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
    15241 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
    15301 ttaagggatt ttggtcatgc atgatatatc tcccaatttg tgtagggctt attatgcacg
    15361 cttaaaaata ataaaagcag acttgacctg at
//
