LOCUS       sequence-182760-       11685 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  protoplast vector for targeted deletion of MLO gene in barley 
            (PvUbi1 promoter driving the gRNA array).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11685)
  AUTHORS   Cermak T, Curtin SJ, Gil-Humanes J, Cegan R, Kono TJY, Konecna E, 
            Belanto JJ, Starker CG, Mathre JW, Greenstein RL, Voytas DF
  TITLE     A multi-purpose toolkit to enable advanced genome engineering in 
            plants.
  JOURNAL   Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 
            10.1105/tpc.16.00922.
  PUBMED    28522548
REFERENCE   2  (bases 1 to 11685)
  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 Cell.
            2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922."
COMMENT     Sequence Label: pTC441
FEATURES             Location/Qualifiers
     source          1..11685
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      84..672
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     573..592
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     826..843
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             3477..3533
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3534..7637
                     /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             3534..7637
                     /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             7638..7685
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      7696..7944
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     CDS             complement(8680..8691)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    9958..9977
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     misc_RNA        9998..10073
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    10074..10093
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     misc_RNA        10114..10189
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    10190..10209
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     polyA_signal    10241..10415
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             10885..11676
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
ORIGIN
        1 cacccatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc
       61 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
      121 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
      181 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
      241 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
      301 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
      361 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
      421 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
      481 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
      541 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
      601 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
      661 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
      721 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
      781 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
      841 gaggaagcgg gagagcgccc atatgcgcac tcctcgcatg cggcgcgccg atctgcaggc
      901 gcgcctgcag tgcagcgtga cccggtcgtg cccctctcta gagataatga gcattgcatg
      961 tctaagttat aaaaaattac cacatatttt ttttgtcaca cttgtttgaa gtgcagttta
     1021 tctatcttta tacatatatt taaactttac tctacgaata atataatcta tagtactaca
     1081 ataatatcag tgttttagag aatcatataa atgaacagtt agacatggtc taaaggacaa
     1141 ttgagtattt tgacaacagg actctacagt tttatctttt tagtgtgcat gtgttctcct
     1201 ttttttttgc aaatagcttc acctatataa tacttcatcc attttattag tacatccatt
     1261 tagggtttag ggttaatggt ttttatagac taattttttt agtacatcta ttttattcta
     1321 ttttagcctc taaattaaga aaactaaaac tctattttag tttttttatt taataattta
     1381 gatataaaat agaataaaat aaagtgacta aaaattaaac aaataccctt taagaaatta
     1441 aaaaaactaa ggaaacattt ttcttgtttc gagtagataa tgccagcctg ttaaacgccg
     1501 tcgacgagtc taacggacac caaccagcga accagcagcg tcgcgtcggg ccaagcgaag
     1561 cagacggcac ggcatctctg tcgctgcctc tggacccctc tcgagagttc cgctccaccg
     1621 ttggacttgc tccgctgtcg gcatccagaa attgcgtggc ggagcggcag acgtgagccg
     1681 gcacggcagg cggcctcctc ctcctctcac ggcaccggca gctacggggg attcctttcc
     1741 caccgctcct tcgctttccc ttcctcgccc gccgtaataa atagacaccc cctccacacc
     1801 ctctttcccc aacctcgtgt tgttcggagc gcacacacac acaaccagat ctcccccaaa
     1861 tccacccgtc ggcacctccg cttcaaggta cgccgctcgt cctccccccc ccccctctct
     1921 accttctcta gatcggcgtt ccggtccatg gttagggccc ggtagttcta cttctgttca
     1981 tgtttgtgtt agatccgtgt ttgtgttaga tccgtgctgc tagcgttcgt acacggatgc
     2041 gacctgtacg tcagacacgt tctgattgct aacttgccag tgtttctctt tggggaatcc
     2101 tgggatggct ctagccgttc cgcagacggg atcgatttca tgattttttt tgtttcgttg
     2161 catagggttt ggtttgccct tttcctttat ttcaatatat gccgtgcact tgtttgtcgg
     2221 gtcatctttt catgcttttt tttgtcttgg ttgtgatgat gtggtctggt tgggcggtcg
     2281 ttctagatcg gagtagaatt aattctgttt caaactacct ggtggattta ttaattttgg
     2341 atctgtatgt gtgtgccata catattcata gttacgaatt gaagatgatg gatggaaata
     2401 tcgatctagg ataggtatac atgttgatgc gggttttact gatgcatata cagagatgct
     2461 ttttgttcgc ttggttgtga tgatgtggtg tggttgggcg gtcgttcatt cgttctagat
     2521 cggagtagaa tactgtttca aactacctgg tgtatttatt aattttggaa ctgtatgtgt
     2581 gtgtcataca tcttcatagt tacgagttta agatggatgg aaatatcgat ctaggatagg
     2641 tatacatgtt gatgtgggtt ttactgatgc atatacatga tggcatatgc agcatctatt
     2701 catatgctct aaccttgagt acctatctat tataataaac aagtatgttt tataattatt
     2761 ttgatcttga tatacttgga tgatggcata tgcagcagct atatgtggat ttttttagcc
     2821 ctgccttcat acgctattta tttgcttggt actgtttctt ttgtcgatgc tcaccctgtt
     2881 gtttggtgtt acttcctgca ggaagcatgg accactacct cgacatcagg ctcaggccag
     2941 acccagagtt cccaccagcc cagctcatgt ccgtcctctt cggcaagctc caccaggccc
     3001 tcgtggccca gggcggcgac aggatcggcg tgtccttccc agacctcgac gagtccaggt
     3061 ccaggctcgg cgagaggctc cgcatccacg cctccgccga cgacctcagg gccctcctcg
     3121 ccaggccgtg gctggagggc ctcagggacc acctccagtt cggcgagcca gccgtggtgc
     3181 cacacccaac cccatacagg caagtgtcca gggtgcaagc caagtccaac ccagagaggc
     3241 tcaggaggag gctcatgagg aggcacgacc tctccgagga agaggccagg aagcgcatcc
     3301 cagacaccgt ggccagggcc ctcgacctcc cattcgtgac cctcaggtcc cagtccaccg
     3361 gccagcactt ccgcctcttc atcaggcacg gcccactcca ggtgaccgcc gaggagggcg
     3421 gctttacctg ctacggcctc tccaagggcg gcttcgtgcc gtggttcggc tccggcgcca
     3481 ccaacttctc cctcctcaag caagccggcg acgtggagga gaacccaggc ccaatggaca
     3541 agaagtactc gatcggcctc gacatcggga cgaactcagt tggctgggcc gtgatcaccg
     3601 acgagtacaa ggtgccctct aagaagttca aggtcctggg gaacaccgac cgccattcca
     3661 tcaagaagaa cctcatcggc gctctcctgt tcgacagcgg ggagaccgct gaggctacga
     3721 ggctcaagag aaccgctagg cgccggtaca cgagaaggaa gaacaggatc tgctacctcc
     3781 aagagatttt ctccaacgag atggccaagg ttgacgattc attcttccac cgcctggagg
     3841 agtctttcct cgtggaggag gataagaagc acgagcggca tcccatcttc ggcaacatcg
     3901 tggacgaggt tgcctaccac gagaagtacc ctacgatcta ccatctgcgg aagaagctcg
     3961 tggactccac cgataaggcg gacctcagac tgatctacct cgctctggcc cacatgatca
     4021 agttccgcgg ccatttcctg atcgaggggg atctcaaccc agacaacagc gatgttgaca
     4081 agctgttcat ccaactcgtg cagacctaca accaactctt cgaggagaac ccgatcaacg
     4141 cctctggcgt ggacgcgaag gctatcctgt ccgcgaggct ctcgaagtcc aggaggctgg
     4201 agaacctgat cgctcagctc ccaggcgaga agaagaacgg cctgttcggg aacctcatcg
     4261 ctctcagcct ggggctcacc ccgaacttca agtcgaactt cgatctcgct gaggacgcca
     4321 agctgcaact ctccaaggac acctacgacg atgacctcga taacctcctg gcccagatcg
     4381 gcgatcaata cgcggacctg ttcctcgctg ccaagaacct gtcggacgcc atcctcctgt
     4441 cagatatcct ccgcgtgaac accgagatca cgaaggctcc actctctgcc tccatgatca
     4501 agcgctacga cgagcaccat caggatctga ccctcctgaa ggcgctggtc cgccaacagc
     4561 tcccggagaa gtacaaggag attttcttcg atcagtcgaa gaacggctac gctgggtaca
     4621 tcgacggcgg ggcctcacaa gaggagttct acaagttcat caagccaatc ctggagaaga
     4681 tggacggcac ggaggagctc ctggtgaagc tcaacaggga ggacctcctg cggaagcaga
     4741 gaaccttcga taacggcagc atcccccacc aaatccatct cggggagctg cacgccatcc
     4801 tgagaaggca agaggacttc taccctttcc tcaaggataa ccgggagaag atcgagaaga
     4861 tcctgacctt cagaatccca tactacgtcg gccctctcgc gcgggggaac tcaagattcg
     4921 cttggatgac ccgcaagtct gaggagacca tcacgccgtg gaacttcgag gaggtggtgg
     4981 acaagggcgc tagcgctcag tcgttcatcg agaggatgac caacttcgac aagaacctgc
     5041 ccaacgagaa ggtgctccct aagcactcgc tcctgtacga gtacttcacc gtctacaacg
     5101 agctcacgaa ggtgaagtac gtcaccgagg gcatgcgcaa gccagcgttc ctgtccgggg
     5161 agcagaagaa ggctatcgtg gacctcctgt tcaagaccaa ccggaaggtc acggttaagc
     5221 aactcaagga ggactacttc aagaagatcg agtgcttcga ttcggtcgag atcagcggcg
     5281 ttgaggaccg cttcaacgcc agcctcggga cctaccacga tctcctgaag atcatcaagg
     5341 ataaggactt cctggacaac gaggagaacg aggatatcct ggaggacatc gtgctgaccc
     5401 tcacgctgtt cgaggacagg gagatgatcg aggagcgcct gaagacgtac gcccatctct
     5461 tcgatgacaa ggtcatgaag caactcaagc gccggagata caccggctgg gggaggctgt
     5521 cccgcaagct catcaacggc atccgggaca agcagtccgg gaagaccatc ctcgacttcc
     5581 tcaagagcga tggcttcgcc aacaggaact tcatgcaact gatccacgat gacagcctca
     5641 ccttcaagga ggatatccaa aaggctcaag tgagcggcca gggggactcg ctgcacgagc
     5701 atatcgcgaa cctcgctggc tcccccgcga tcaagaaggg catcctccag accgtgaagg
     5761 ttgtggacga gctcgtgaag gtcatgggcc ggcacaagcc tgagaacatc gtcatcgaga
     5821 tggccagaga gaaccaaacc acgcagaagg ggcaaaagaa ctctagggag cgcatgaagc
     5881 gcatcgagga gggcatcaag gagctggggt cccaaatcct caaggagcac ccagtggaga
     5941 acacccaact gcagaacgag aagctctacc tgtactacct ccagaacggc agggatatgt
     6001 acgtggacca agagctggat atcaaccgcc tcagcgatta cgacgtcgat catatcgttc
     6061 cccagtcttt cctgaaggat gactccatcg acaacaaggt cctcaccagg tcggacaaga
     6121 accgcggcaa gtcagataac gttccatctg aggaggtcgt taagaagatg aagaactact
     6181 ggaggcagct cctgaacgcc aagctgatca cgcaaaggaa gttcgacaac ctcaccaagg
     6241 ctgagagagg cgggctctca gagctggaca aggccggctt catcaagcgg cagctggtcg
     6301 agaccagaca aatcacgaag cacgttgcgc aaatcctcga ctctcggatg aacacgaagt
     6361 acgatgagaa cgacaagctg atcagggagg ttaaggtgat caccctgaag tctaagctcg
     6421 tctccgactt caggaaggat ttccagttct acaaggttcg cgagatcaac aactaccacc
     6481 atgcccatga cgcttacctc aacgctgtgg tcggcaccgc tctgatcaag aagtacccaa
     6541 agctggagtc cgagttcgtg tacggggact acaaggttta cgatgtgcgc aagatgatcg
     6601 ccaagtcgga gcaagagatc ggcaaggcta ccgccaagta cttcttctac tcaaacatca
     6661 tgaacttctt caagaccgag atcacgctgg ccaacggcga gatccggaag agaccgctca
     6721 tcgagaccaa cggcgagacg ggggagatcg tgtgggacaa gggcagggat ttcgcgaccg
     6781 tccgcaaggt tctctccatg ccccaggtga acatcgtcaa gaagaccgag gtccaaacgg
     6841 gcgggttctc aaaggagtct atcctgccta agcggaacag cgacaagctc atcgccagaa
     6901 agaaggactg ggacccaaag aagtacggcg ggttcgacag ccctaccgtg gcctactcgg
     6961 tcctggttgt ggcgaaggtt gagaagggca agtccaagaa gctcaagagc gtgaaggagc
     7021 tcctggggat caccatcatg gagaggtcca gcttcgagaa gaacccaatc gacttcctgg
     7081 aggccaaggg ctacaaggag gtgaagaagg acctgatcat caagctcccg aagtactctc
     7141 tcttcgagct ggagaacggc aggaagagaa tgctggcttc cgctggcgag ctccagaagg
     7201 ggaacgagct cgcgctgcca agcaagtacg tgaacttcct ctacctggct tcccactacg
     7261 agaagctcaa gggcagcccg gaggacaacg agcaaaagca gctgttcgtc gagcagcaca
     7321 agcattacct cgacgagatc atcgagcaaa tctccgagtt cagcaagcgc gtgatcctcg
     7381 ccgacgcgaa cctggataag gtcctctccg cctacaacaa gcaccgggac aagcccatca
     7441 gagagcaagc ggagaacatc atccatctct tcaccctgac gaacctcggc gctcctgctg
     7501 ctttcaagta cttcgacacc acgatcgatc ggaagagata cacctccacg aaggaggtcc
     7561 tggacgcgac cctcatccac cagtcgatca ccggcctgta cgagacgagg atcgacctct
     7621 cacaactcgg cggggataag agacccgcag caaccaagaa ggcagggcaa gcaaagaaga
     7681 agaagtgact cgagatatga agatgaagat gaaatatttg gtgtgtcaaa taaaaagctt
     7741 gtgtgcttaa gtttgtgttt ttttcttggc ttgttgtgtt atgaatttgt ggctttttct
     7801 aatattaaat gaatgtaaga tcacattata atgaataaac aaatgtttct ataatccatt
     7861 gtgaatgttt tgttggatct cttctgcagc atataactac tgtatgtgct atggtatgga
     7921 ctatggaata tgattaaaga taaggagctc cggtgacgga cggcgcgccg aagagcacgt
     7981 cagtgtttgg tttccactag cacgagtagc gcaatcagaa aattttcaat gcatgaagta
     8041 ctaaacgaag tttatttaga aattttttta agaaatgagt gtaatttttt gcgacgaatt
     8101 taatgacaat aattaatcga tgattgccta cagtaatgct acagtaacca acctctaatc
     8161 atgcgtcgaa tgcgtcatta gattcgtctc gcaaaatagc acaagaatta tgaaattaat
     8221 tttacaaact atttttattt aatactaata attaactgtc aaagtttgtg ctactcgcaa
     8281 gagtagcgcg aaccaaacac ggcctggagg agcacggtaa cggcgtcgac aaactaacgg
     8341 ccaccacccg ccaacgcaaa ggagacggat gagagttgac ttcttgacgg ttctccaccc
     8401 ctctgtctct ctgtcactgg gccctgggtc cccctctcga aagttcctct ggccgaaatt
     8461 gcgcggcgga gacgaggcgg gcggaaccgt cacggcagag gattccttcc ccaccctgcc
     8521 tggcccggcc atatataaac agccaccgcc cctccccgtt ccccatcgcg tctcgtctcg
     8581 tgttgttccc agaacacaac caaaatccaa atcctcctcc tcctcccgag cctcgtcgat
     8641 ccctcacccg cttcaaggta cggcgatcct cctctccctt ctcccctcga tcgattatgc
     8701 gtgttccgtt tccgtttccg atcgagcgaa tcgatggtta ggacccatgg gggacccatg
     8761 gggtgtcgtg tggtggtctg gtttgatccg cgatatttct ccgttcgtag tgtagatctg
     8821 atcgaatccc tggtgaaatc gttgatcgtg ctattcgtgt gagggttctt aggtttggag
     8881 ttgtggaggt agttctgatc ggtttgtagg tgagattttc cccatgattt tgcttggctc
     8941 gtttgtcttg gttagattag atctgcccgc attttgttcg atatttctga tgcagatatg
     9001 atgaataatt tcgtccttgt atcccgcgtc cgtatgtgta ttaagtttgc aggtcctagt
     9061 taggtttttc ctactgattt gtcttatcca ttctgtttag cttgcaaggt ttggtaatgg
     9121 tccggcatgt ttgtctctat agattagagt agaataagat tatctcaaca agctgttggc
     9181 ttatcaattt tggatctgca tgtgtttcgc atctatatct ttgcaattaa gatggtagat
     9241 ggacatatgc tcctgttgag ttgatgttgt accttttacc tgaggtctga ggaacatgca
     9301 tcctcctgct actttgtgct tatacagatc atcaagatta tgtagctaat attcgatcag
     9361 tttctagtat ctacatggta aacttgcatg cacttgctac ttatttttga tatacttgga
     9421 tgataacata tgctgctggt tgattcctac ctacatgatg aacattttac aggccattag
     9481 tgtctgtctg tatgtgttgt tcctgtttgc ttcagtctat ttctgtttca ttcctagttt
     9541 attggttctc tgctagatac ttaccctgct gggcttagtt atcatcttat ctcgaatgca
     9601 ttttcatgtt tatagatgaa tatacactca gataggtgta gatgtatgct actgtttctc
     9661 tacgttgctg taggttttac ctgtggcaac tgcatactcc tgttgcttcg ctagatatgt
     9721 atgtgcttat atagattaag atatgtgtga tggttcttta gtatatctga tgatcatgta
     9781 tgctctttta acttcttgct acacttggta acatgctgtg atgctgtttg ttgattctgt
     9841 agcactacca atgatgacct tatctctctt tgtatatgat gtttctgttt gtttgaggct
     9901 tgtgttactg ctagttactt accctgttgc ctggctaatc ttctgcagat cctgcaggtt
     9961 cactgccgta taggcaggac ggacacgagc accatgggtt ttagagctag aaatagcaag
    10021 ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcgttcact
    10081 gccgtatagg cagtaaatcc taacgacagg agggttttag agctagaaat agcaagttaa
    10141 aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgcg ttcactgccg
    10201 tataggcagg tcagaagagc agtcgatcga caagctcgag tttctccata ataatgtgtg
    10261 agtagttccc agataaggga attagggttc ctatagggtt tcgctcatgt gttgagcata
    10321 taagaaaccc ttagtatgta tttgtatttg taaaatactt ctatcaataa aatttctaat
    10381 tcctaaaacc aaaatccagt actaaaatcc agatcccccg aattaccggc gagctttggg
    10441 tacgtcacgt ggctcgagcg cgtagtcctc ggtaatatcg cagaacaaaa gtacctgata
    10501 tcgagtgtac ttcaagtcac accggcgagt gccagccagg acagaaatgc ctcgacttcg
    10561 ctgctaccca aggttgccgg gtgacgcaca ccgtggaaac ggatgaaggc acgaacccag
    10621 tggacataag cctgttcggt tcgtaagctg taatgcaagt agcgtatgcg ctcacgcaac
    10681 tggtccagaa ccttgaccga acgcagcggt ggtaacggcg cagtggcggt tttcatggct
    10741 tgttatgact gtttttttgg ggtacagtct atgcctcggg catccaagca gcaagcgcgt
    10801 tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg atgttacgca gcagggcagt
    10861 cgccctaaaa caaagttaaa cattatgagg gaagcggtga tcgccgaagt atcgactcaa
    10921 ctatcagagg tagttggcgt catcgagcgc catctcgaac cgacgttgct ggccgtacat
    10981 ttgtacggct ccgcagtgga tggcggcctg aagccacaca gtgatattga tttgctggtt
    11041 acggtgaccg taaggcttga tgaaacaacg cggcgagctt tgatcaacga ccttttggaa
    11101 acttcggctt cccctggaga gagcgagatt ctccgcgctg tagaagtcac cattgttgtg
    11161 cacgacgaca tcattccgtg gcgttatcca gctaagcgcg aactgcaatt tggagaatgg
    11221 cagcgcaatg acattcttgc aggtatcttc gagccagcca cgatcgacat tgatctggct
    11281 atcttgctga caaaagcaag agaacatagc gttgccttgg taggtccagc ggcggaggaa
    11341 ctctttgatc cggttcctga acaggatcta tttgaggcgc taaatgaaac cttaacgcta
    11401 tggaactcgc cgcccgactg ggctggcgat gagcgaaatg tagtgcttac gttgtcccgc
    11461 atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc cgactgggca
    11521 atggagcgcc tgccggccca gtatcagccc gtcatacttg aagctagaca ggcttatctt
    11581 ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt tgtccactac
    11641 gtgaaaggcg agatcaccaa ggtagtcggc aaataaccct cgagc
//
