LOCUS       sequence-337777-       13805 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express SpCas9 and Basta resistance gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13805)
  AUTHORS   Oh Y, Lee B, Kim H, Kim SG
  TITLE     A multiplex guide RNA expression system and its efficacy for plant 
            genome engineering.
  JOURNAL   Plant Methods. 2020 Mar 12;16:37. doi: 10.1186/s13007-020-00580-x. 
            eCollection 2020.
  PUBMED    32190101
REFERENCE   2  (bases 1 to 13805)
  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: "Plant 
            Methods."; date: "2020-03-12"; volume: "16:37. doi"; pages: " 
            10.1186/s13007-020-00580-x. eCollection 2020"
COMMENT     Sequence Label: pECO200
FEATURES             Location/Qualifiers
     source          1..13805
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        complement(1..76)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        993..1337
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     1281..1301
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     protein_bind    1437..1461
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1464..5567
                     /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             1464..5567
                     /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             5583..5603
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5604..5630
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             5604..5630
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(5635..5659)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    5979..6003
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             7334..7963
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             8392..9465
                     /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      9531..9725
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(9961..9983)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    10069..10209
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     10124..10143
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(10224..10241)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10395..10983)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10475..10494)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(11229..12020)
                     /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    12541..12565
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(12658..12910)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(12930..13481)
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     promoter        complement(13523..13702)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
ORIGIN
        1 gcaccgactc ggtgccactt tttcaagttg ataacggact agccttattt taacttgcta
       61 tttctagctc taaaacctct gcaggtgact ctcgagacac ctgcctccaa tcactacttc
      121 gactctagct gtatataaac tcagcttcgt tttcttatct aagcgatgtg ggacttttga
      181 agattgtttt caacttaaat gggcctatat aagaaatact attgttcttt cccatataaa
      241 tgggcctgct tctcttcttt cagattccca ggggcctttt gaagattatc ttcatatctt
      301 aagaatgaag atgttttatt caatcaaatt cttgaaggtt cgatgcctaa tcattcgaat
      361 tcgagctctc ccatatggtc gactagagcc aagctgatct cctttgcccc ggagatcacc
      421 atggacgact ttctctatct ctacgatcta ggaagaaagt tcgacggaga aggtgacgat
      481 accatgttca ccaccgataa tgagaagatt agcctcttca atttcagaaa gaatgctgac
      541 ccacagatgg ttagagaggc ctacgcggca ggtctcatca agacgatcta cccgagtaat
      601 aatctccagg agatcaaata ccttcccaag aaggttaaag atgcagtcaa aagattcagg
      661 actaactgca tcaagaacac agagaaagat atatttctca agatcagaag tactattcca
      721 gtatggacga ttcaaggctt gcttcataaa ccaaggcaag taatagagat tggagtctct
      781 aagaaagtag ttcctactga atcaaaggcc atggagtcaa aaattcagat cgaggatcta
      841 acagaactcg ccgtgaagac tggcgaacag ttcatacaga gtcttttacg actcaatgac
      901 aagaagaaaa tcttcgtcaa catggtggag cacgacactc tcgtctactc caagaatatc
      961 aaagatacag tctcagaaga ccaaagggct attgagactt ttcaacaaag ggtaatatcg
     1021 ggaaacctcc tcggattcca ttgcccagct atctgtcact tcatcaaaag gacagtagaa
     1081 aaggaaggtg gcacctacaa atgccatcat tgcgataaag gaaaggctat cgttcaagat
     1141 gcccctgccg acagtggtcc caaagatgga cccccaccca cgaggagcat cgtggaaaaa
     1201 gaagacgttc caaccacgtc ttcaaagcaa gtggattgat gtgatatctc cactgacgta
     1261 agggatgacg cacaatccca ctatccttcg caagaccctt cctctatata aggaagttca
     1321 tttcatttgg agaggactcc ggtattttta caacaattac cacaacaaaa caaacaacaa
     1381 acaacattac aatttactat tctagtcgac ctgcaggcgg ccgcactagt gatatcacaa
     1441 gtttgtacaa aaaagcaggc ttcatggaca agaagtacag catcggcctg gacatcggta
     1501 ccaacagcgt gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc aagaagttca
     1561 aggtgctggg caacaccgac cgccacagca tcaagaagaa cctgatcggc gccctgctgt
     1621 tcgacagcgg cgagaccgcc gaggccaccc gcctgaagcg caccgcccgc cgccgctaca
     1681 cccgccgcaa gaaccgcatc tgctacctgc aggagatctt cagcaacgag atggccaagg
     1741 tggacgacag cttcttccac cgcctggagg agagcttcct ggtggaggag gacaagaagc
     1801 acgagcgcca ccccatcttc ggcaacatcg tggacgaggt ggcctaccac gagaagtacc
     1861 ccaccatcta ccatctgcgc aagaagctgg tggacagcac cgacaaggcc gacctgcgcc
     1921 tgatctacct ggccctggcc cacatgatca agttccgcgg ccacttcctg atcgagggcg
     1981 acctgaaccc cgacaacagc gacgtggaca agctgttcat ccagctggtg cagacctaca
     2041 accagctgtt cgaggagaac cccatcaacg ccagcggcgt ggacgccaag gccatcctga
     2101 gcgcccgcct gagcaagagc cgccgcctgg agaacctgat cgcccagctg cccggcgaga
     2161 agaagaacgg cctgttcggc aacctgatcg ccctgagcct gggcctgacc cccaacttca
     2221 agagcaactt cgacctggcc gaggacgcca agctgcagct gagcaaggac acctacgacg
     2281 acgacctgga caacctgctg gcccagatcg gcgaccagta cgccgacctg ttcctggccg
     2341 ccaagaacct gagcgacgcc atcctgctga gcgacatcct gcgcgtgaac accgagatca
     2401 ccaaggcccc cctgagcgcc agcatgatca agcgctacga cgagcaccac caggacctga
     2461 ccctgctgaa ggccctggtg cgccagcagc tgcccgagaa gtacaaggag atcttcttcg
     2521 accagagcaa gaacggctac gccggctaca tcgacggcgg cgccagccag gaggagttct
     2581 acaagttcat caagcccatc ctggagaaga tggacggcac cgaggagctg ctggtgaagc
     2641 tgaaccgcga ggacctgctg cgcaagcagc gcaccttcga caacggcagc atcccccacc
     2701 agatccacct gggcgagctg cacgccatcc tgcgccgcca ggaggacttc taccccttcc
     2761 tgaaggacaa ccgcgagaag atcgagaaga tcctgacctt ccgcatcccc tactacgtgg
     2821 gccccctggc ccgcggcaac agccgcttcg cctggatgac ccgcaagagc gaggagacca
     2881 tcaccccctg gaacttcgag gaggtggtgg acaagggcgc cagcgcccag agcttcatcg
     2941 agcgcatgac caacttcgac aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc
     3001 tgctgtacga gtacttcacc gtgtacaacg agctgaccaa ggtgaagtac gtgaccgagg
     3061 gcatgcgcaa gcccgccttc ctgagcggcg agcagaagaa ggccatcgtg gacctgctgt
     3121 tcaagaccaa ccgcaaggtg accgtgaagc agctgaagga ggactacttc aagaagatcg
     3181 agtgcttcga cagcgtggag atcagcggcg tggaggaccg cttcaacgcc agcctgggca
     3241 cctaccacga cctgctgaag atcatcaagg acaaggactt cctggacaac gaggagaacg
     3301 aggacatcct ggaggacatc gtgctgaccc tgaccctgtt cgaggaccgc gagatgatcg
     3361 aggagcgcct gaagacctac gcccacctgt tcgacgacaa ggtgatgaag cagctgaagc
     3421 gccgccgcta caccggctgg ggccgcctga gccgcaagct tatcaacggc atccgcgaca
     3481 agcagagcgg caagaccatc ctggacttcc tgaagagcga cggcttcgcc aaccgcaact
     3541 tcatgcagct gatccacgac gacagcctga ccttcaagga ggacatccag aaggcccagg
     3601 tgagcggcca gggcgacagc ctgcacgagc acatcgccaa cctggccggc agccccgcca
     3661 tcaagaaggg catcctgcag accgtgaagg tggtggacga gctggtgaag gtgatgggcc
     3721 gccacaagcc cgagaacatc gtgatcgaga tggcccgcga gaaccagacc acccagaagg
     3781 gccagaagaa cagccgcgag cgcatgaagc gcatcgagga gggcatcaag gagctgggca
     3841 gccagatcct gaaggagcac cccgtggaga acacccagct gcagaacgag aagctgtacc
     3901 tgtactacct gcagaacggc cgcgacatgt acgtggacca ggagctggac atcaaccgcc
     3961 tgagcgacta cgacgtggac cacatcgtgc cccagagctt cctgaaggac gacagcatcg
     4021 acaacaaggt gctgacccgc agcgacaaga accgcggcaa gagcgacaac gtgcccagcg
     4081 aggaggtggt gaagaagatg aagaactact ggcgccagct gctgaacgcc aagctgatca
     4141 cccagcgcaa gttcgacaac ctgaccaagg ccgagcgcgg cggcctgagc gagctggaca
     4201 aggccggctt catcaagcgc cagctggtgg agacccgcca gatcaccaag cacgtggccc
     4261 agatcctgga cagccgcatg aacaccaagt acgacgagaa cgacaagctg atccgcgagg
     4321 tgaaggtgat caccctgaag agcaagctgg tgagcgactt ccgcaaggac ttccagttct
     4381 acaaggtgcg cgagatcaac aactaccacc acgcccacga cgcctacctg aacgccgtgg
     4441 tgggcaccgc cctgatcaag aagtacccca agctggagag cgagttcgtg tacggcgact
     4501 acaaggtgta cgacgtgcgc aagatgatcg ccaagagcga gcaggagatc ggcaaggcca
     4561 ccgccaagta cttcttctac agcaacatca tgaacttctt caagaccgag atcaccctgg
     4621 ccaacggcga gatccgcaag cgccccctga tcgagaccaa cggcgagacc ggcgagatcg
     4681 tgtgggacaa gggccgcgac ttcgccaccg tgcgcaaggt gctgagcatg ccccaggtga
     4741 acatcgtgaa gaagaccgag gtgcagaccg gcggcttcag caaggagagc atcctgccca
     4801 agcgcaacag cgacaagctg atcgcccgca agaaggactg ggaccccaag aagtacggcg
     4861 gcttcgacag ccccaccgtg gcctacagcg tgctggtggt ggccaaggtg gagaagggca
     4921 agagcaagaa gctgaagagc gtgaaggagc tgctgggcat caccatcatg gagcgcagca
     4981 gcttcgagaa gaaccccatc gacttcctgg aggccaaggg ctacaaggag gtgaagaagg
     5041 acctgatcat caagctgccc aagtacagcc tgttcgagct ggagaacggc cgcaagcgca
     5101 tgctggccag cgccggcgag ctgcagaagg gcaacgagct ggccctgccc agcaagtacg
     5161 tgaacttcct gtacctggcc agccactacg agaagctgaa gggcagcccc gaggacaacg
     5221 agcagaagca gctgttcgtg gagcagcaca agcactacct ggacgagatc atcgagcaga
     5281 tcagcgagtt cagcaagcgc gtgatcctgg ccgacgccaa cctggacaag gtgctgagcg
     5341 cctacaacaa gcaccgcgac aagcccatcc gcgagcaggc cgagaacatc atccacctgt
     5401 tcaccctgac caacctgggc gcccccgccg ccttcaagta cttcgacacc accatcgacc
     5461 gcaagcgcta caccagcacc aaggaggtgc tggacgccac cctgatccac cagagcatca
     5521 ccggtctgta cgagacccgc atcgacctga gccagctggg cggcgacggc ggctccggac
     5581 ctccaaagaa aaagagaaaa gtatacccct acgacgtgcc cgactacgcc taacacccag
     5641 ctttcttgta caaagtggtg atatcccgcg gccatgctag agtccgcaaa aatcaccagt
     5701 ctctctctac aaatctatct ctctctattt ttctccagaa taatgtgtga gtagttccca
     5761 gataagggaa ttagggttct tatagggttt cgctcatgtg ttgagcatat aagaaaccct
     5821 tagtatgtat ttgtatttgt aaaatacttc tatcaataaa atttctaatt cctaaaacca
     5881 aaatccagtg acctgcaggc atgcgacgtc gggcccaagc ttagcttgag cttggatcag
     5941 attgtcgttt cccgccttca gtttaaacta tcagtgtttg acaggatata ttggcgggta
     6001 aacctaagag aaaagagcgt ttattagaat aatcggatat ttaaaagggc gtgaaaaggt
     6061 ttatccgttc gtccatttgt atgtgcatgc caaccacagg gttcccctcg ggatcaaagt
     6121 actttaaagt actttaaagt actttaaagt actttgatcc aacccctccg ctgctatagt
     6181 gcagtcggct tctgacgttc agtgcagccg tcttctgaaa acgacatgtc gcacaagtcc
     6241 taagttacgc gacaggctgc cgccctgccc ttttcctggc gttttcttgt cgcgtgtttt
     6301 agtcgcataa agtagaatac ttgcgactag aaccggagac attacgccat gaacaagagc
     6361 gccgccgctg gcctgctggg ctatgcccgc gtcagcaccg acgaccagga cttgaccaac
     6421 caacgggccg aactgcacgc ggccggctgc accaagctgt tttccgagaa gatcaccggc
     6481 accaggcgcg accgcccgga gctggccagg atgcttgacc acctacgccc tggcgacgtt
     6541 gtgacagtga ccaggctaga ccgcctggcc cgcagcaccc gcgacctact ggacattgcc
     6601 gagcgcatcc aggaggccgg cgcgggcctg cgtagcctgg cagagccgtg ggccgacacc
     6661 accacgccgg ccggccgcat ggtgttgacc gtgttcgccg gcattgccga gttcgagcgt
     6721 tccctaatca tcgaccgcac ccggagcggg cgcgaggccg ccaaggcccg aggcgtgaag
     6781 tttggccccc gccctaccct caccccggca cagatcgcgc acgcccgcga gctgatcgac
     6841 caggaaggcc gcaccgtgaa agaggcggct gcactgcttg gcgtgcatcg ctcgaccctg
     6901 taccgcgcac ttgagcgcag cgaggaagtg acgcccaccg aggccaggcg gcgcggtgcc
     6961 ttccgtgagg acgcattgac cgaggccgac gccctggcgg ccgccgagaa tgaacgccaa
     7021 gaggaacaag catgaaaccg caccaggacg gccaggacga accgtttttc attaccgaag
     7081 agatcgaggc ggagatgatc gcggccgggt acgtgttcga gccgcccgcg cacgtctcaa
     7141 ccgtgcggct gcatgaaatc ctggccggtt tgtctgatgc caagctggcg gcctggccgg
     7201 ccagcttggc cgctgaagaa accgagcgcc gccgtctaaa aaggtgatgt gtatttgagt
     7261 aaaacagctt gcgtcatgcg gtcgctgcgt atatgatgcg atgagtaaat aaacaaatac
     7321 gcaaggggaa cgcatgaagg ttatcgctgt acttaaccag aaaggcgggt caggcaagac
     7381 gaccatcgca acccatctag cccgcgccct gcaactcgcc ggggccgatg ttctgttagt
     7441 cgattccgat ccccagggca gtgcccgcga ttgggcggcc gtgcgggaag atcaaccgct
     7501 aaccgttgtc ggcatcgacc gcccgacgat tgaccgcgac gtgaaggcca tcggccggcg
     7561 cgacttcgta gtgatcgacg gagcgcccca ggcggcggac ttggctgtgt ccgcgatcaa
     7621 ggcagccgac ttcgtgctga ttccggtgca gccaagccct tacgacatat gggccaccgc
     7681 cgacctggtg gagctggtta agcagcgcat tgaggtcacg gatggaaggc tacaagcggc
     7741 ctttgtcgtg tcgcgggcga tcaaaggcac gcgcatcggc ggtgaggttg ccgaggcgct
     7801 ggccgggtac gagctgccca ttcttgagtc ccgtatcacg cagcgcgtga gctacccagg
     7861 cactgccgcc gccggcacaa ccgttcttga atcagaaccc gagggcgacg ctgcccgcga
     7921 ggtccaggcg ctggccgctg aaattaaatc aaaactcatt tgagttaatg aggtaaagag
     7981 aaaatgagca aaagcacaaa cacgctaagt gccggccgtc cgagcgcacg cagcagcaag
     8041 gctgcaacgt tggccagcct ggcagacacg ccagccatga agcgggtcaa ctttcagttg
     8101 ccggcggagg atcacaccaa gctgaagatg tacgcggtac gccaaggcaa gaccattacc
     8161 gagctgctat ctgaatacat cgcgcagcta ccagagtaaa tgagcaaatg aataaatgag
     8221 tagatgaatt ttagcggcta aaggaggcgg catggaaaat caagaacaac caggcaccga
     8281 cgccgtggaa tgccccatgt gtggaggaac gggcggttgg ccaggcgtaa gcggctgggt
     8341 tgtctgccgg ccctgcaatg gcactggaac ccccaagccc gaggaatcgg cgtgagcggt
     8401 cgcaaaccat ccggcccggt acaaatcggc gcggcgctgg gtgatgacct ggtggagaag
     8461 ttgaaggccg cgcaggccgc ccagcggcaa cgcatcgagg cagaagcacg ccccggtgaa
     8521 tcgtggcaag cggccgctga tcgaatccgc aaagaatccc ggcaaccgcc ggcagccggt
     8581 gcgccgtcga ttaggaagcc gcccaagggc gacgagcaac cagatttttt cgttccgatg
     8641 ctctatgacg tgggcacccg cgatagtcgc agcatcatgg acgtggccgt tttccgtctg
     8701 tcgaagcgtg accgacgagc tggcgaggtg atccgctacg agcttccaga cgggcacgta
     8761 gaggtttccg cagggccggc cggcatggcc agtgtgtggg attacgacct ggtactgatg
     8821 gcggtttccc atctaaccga atccatgaac cgataccggg aagggaaggg agacaagccc
     8881 ggccgcgtgt tccgtccaca cgttgcggac gtactcaagt tctgccggcg agccgatggc
     8941 ggaaagcaga aagacgacct ggtagaaacc tgcattcggt taaacaccac gcacgttgcc
     9001 atgcagcgta cgaagaaggc caagaacggc cgcctggtga cggtatccga gggtgaagcc
     9061 ttgattagcc gctacaagat cgtaaagagc gaaaccgggc ggccggagta catcgagatc
     9121 gagctagctg attggatgta ccgcgagatc acagaaggca agaacccgga cgtgctgacg
     9181 gttcaccccg attacttttt gatcgatccc ggcatcggcc gttttctcta ccgcctggca
     9241 cgccgcgccg caggcaaggc agaagccaga tggttgttca agacgatcta cgaacgcagt
     9301 ggcagcgccg gagagttcaa gaagttctgt ttcaccgtgc gcaagctgat cgggtcaaat
     9361 gacctgccgg agtacgattt gaaggaggag gcggggcagg ctggcccgat cctagtcatg
     9421 cgctaccgca acctgatcga gggcgaagca tccgccggtt cctaatgtac ggagcagatg
     9481 ctagggcaaa ttgccctagc aggggaaaaa ggtcgaaaag gtctctttcc tgtggatagc
     9541 acgtacattg ggaacccaaa gccgtacatt gggaaccgga acccgtacat tgggaaccca
     9601 aagccgtaca ttgggaaccg gtcacacatg taagtgactg atataaaaga gaaaaaaggc
     9661 gatttttccg cctaaaactc tttaaaactt attaaaactc ttaaaacccg cctggcctgt
     9721 gcataactgt ctggccagcg cacagccgaa gagctgcaaa aagcgcctac ccttcggtcg
     9781 ctgcgctccc tacgccccgc cgcttcgcgt cggcctatcg cggccgctgg ccgctcaaaa
     9841 atggctggcc tacggccagg caatctacca gggcgcggac aagccgcgcc gtcgccactc
     9901 gaccgccggc gcccacatca aggcaccctg cctcgcgcgt ttcggtgatg acggtgaaaa
     9961 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
    10021 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac
    10081 ccagtcacgt agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt
    10141 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
    10201 cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
    10261 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
    10321 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
    10381 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
    10441 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
    10501 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
    10561 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
    10621 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
    10681 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
    10741 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
    10801 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
    10861 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
    10921 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
    10981 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
    11041 taagggattt tggtcatgca tgatatatct cccaatttgt gtagggctta ttatgcacgc
    11101 ttaaaaataa taaaagcaga cttgacctga tagtttggct gtgagcaatt atgtgcttag
    11161 tgcatctaac gcttgagtta agccgcgccg cgaagcggcg tcggcttgaa cgaatttcta
    11221 gctagacatt atttgccgac taccttggtg atctcgcctt tcacgtagtg gacaaattct
    11281 tccaactgat ctgcgcgcga ggccaagcga tcttcttctt gtccaagata agcctgtcta
    11341 gcttcaagta tgacgggctg atactgggcc ggcaggcgct ccattgccca gtcggcagcg
    11401 acatccttcg gcgcgatttt gccggttact gcgctgtacc aaatgcggga caacgtaagc
    11461 actacatttc gctcatcgcc agcccagtcg ggcggcgagt tccatagcgt taaggtttca
    11521 tttagcgcct caaatagatc ctgttcagga accggatcaa agagttcctc cgccgctgga
    11581 cctaccaagg caacgctatg ttctcttgct tttgtcagca agatagccag atcaatgtcg
    11641 atcgtggctg gctcgaagat acctgcaaga atgtcattgc gctgccattc tccaaattgc
    11701 agttcgcgct tagctggata acgccacgga atgatgtcgt cgtgcacaac aatggtgact
    11761 tctacagcgc ggagaatctc gctctctcca ggggaagccg aagtttccaa aaggtcgttg
    11821 atcaaagctc gccgcgttgt ttcatcaagc cttacggtca ccgtaaccag caaatcaata
    11881 tcactgtgtg gcttcaggcc gccatccact gcggagccgt acaaatgtac ggccagcaac
    11941 gtcggttcga gatggcgctc gatgacgcca actacctctg atagttgagt cgatacttcg
    12001 gcgatcaccg cttcccccat gatgtttaac tttgttttag ggcgactgcc ctgctgcgta
    12061 acatcgttgc tgctccataa catcaaacat cgacccacgg cgtaacgcgc ttgctgcttg
    12121 gatgcccgag gcatagactg taccccaaaa aaacagtcat aacaagccat gaaaaccgcc
    12181 actgcgccgt taccaccgct gcgttcggtc aaggttctgg accagttgcg tgagcgcata
    12241 cgctacttgc attacagctt acgaaccgaa caggcttatg tccactgggt tcgtgcccga
    12301 attgatcaca ggcagcaacg ctctgtcatc gttacaatca acatgctacc ctccgcgaga
    12361 tcatccgtgt ttcaaacccg gcagcttagt tgccgttctt ccgaatagca tcggtaacat
    12421 gagcaaagtc tgccgcctta caacggctct cccgctgacg ccgtcccgga ctgatgggct
    12481 gcctgtatcg agtggtgatt ttgtgccgag ctgccggtcg gggagctgtt ggctggctgg
    12541 tggcaggata tattgtggtg taaacaaatt gacgcttaga caacttaata acacattgcg
    12601 gacgttttta atgtactgaa ttaacgccga attgaattat cagcttgcat gccggtcgat
    12661 ctagtaacat agatgacacc gcgcgcgata atttatccta gtttgcgcgc tatattttgt
    12721 tttctatcgc gtattaaatg tataattgcg ggactctaat cataaaaacc catctcataa
    12781 ataacgtcat gcattacatg ttaattatta catgcttaac gtaattcaac agaaattata
    12841 tgataatcat cgcaagaccg gcaacaggat tcaatcttaa gaaactttat tgccaaatgt
    12901 ttgaacgatc tgcttgactc taggggtcat cagatttcgg tgacgggcag gaccggacgg
    12961 ggcggcaccg gcaggctgaa gtccagctgc cagaaaccca cgtcatgcca gttcccgtgc
    13021 ttgaagccgg ccgcccgcag catgccgcgg ggggcatatc cgagcgcctc gtgcatgcgc
    13081 acgctcgggt cgttgggcag cccgatgaca gcgaccacgc tcttgaagcc ctgtgcctcc
    13141 agggacttca gaaggtgggt gtagagcgtg gagcccagtc ccgtccgctg gtggcggggg
    13201 gagacgtaca cggtcgactc ggccgtccag tcgtaggcgt tgcgtgcctt ccagggaccc
    13261 gcgtaggcga tgccggcgac ctcgccgtcc acctcggcga cgagccaggg atagcgctcc
    13321 cgcagacgga cgaggtcgtc cgtccactcc tgcggttcct gcggctcggt acggaagttg
    13381 accgtgcttg tctcgatgta gtggttgacg atggtgcaga ccgccggcat gtccgcctcg
    13441 gtggcacggc ggatgtcggc cgggcgtcgt tctgggctca tggtagatcc cctcgatcga
    13501 gttgagagtg aatatgagac tctaattgga taccgagggg aatttatgga acgtcagtgg
    13561 agcatttttg acaagaaata tttgctagct gatagtgacc ttaggcgact tttgaacgcg
    13621 caataatggt ttctgacgta tgtgcttagc tcattaaact ccagaaaccc gcggctcagt
    13681 ggctccttca acgttgcggt tctgtcagtt ccaaacgtaa aacggcttgt cccgcgtcat
    13741 cggcgggggt cataacgtga ctcccttaat tctccgctca tgatcgataa ttcgagctca
    13801 aaaaa
//
