LOCUS       sequence-101817-       13632 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR/Cas based plant genome editing and gene regulation; expresses
            3×FLAG-NLS-zCas9-NLS, gRNA scaffold for insertion of target sequence
            (TaU3 promoter), Bar resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13632)
  AUTHORS   Xing HL, Dong L, Wang ZP, Zhang HY, Han CY, Liu B, Wang XC, Chen QJ
  TITLE     A CRISPR/Cas9 toolkit for multiplex genome editing in plants.
  JOURNAL   BMC Plant Biol. 2014 Nov 29;14(1):327.
  PUBMED    25432517
REFERENCE   2  (bases 1 to 13632)
  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: "BMC Plant 
            Biol."; date: "2014-11-29"; volume: "14(1)"; pages: "327"
COMMENT     Sequence Label: pBUN421
FEATURES             Location/Qualifiers
     source          1..13632
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..25
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             910..1701
                     /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_RNA        1853..1928
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2263..4253
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     CDS             4299..4364
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4371..4391
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4416..8516
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4416..8516
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     RBS             8429..8437
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             8517..8564
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      8609..8861
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(8870..8887)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8870..8886)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8879..8901)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        9086..9763
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     CDS             9808..10359
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     polyA_signal    10366..10540
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    10690..10714
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     complement(10962..10985)
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     terminator      complement(10993..11020)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(11112..11198)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      11367..11802
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     rep_origin      complement(11917..12505)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11997..12016)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(12595..13404)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVSLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     13318..13337
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     13396..13415
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 gtttacccgc caatatatcc tgtcaaacac tgatagttta aactgaaggc gggaaacgac
       61 aatctgatcc aagctcaagc taagctcacg tgacggaatt aagcttcatg aatccaaacc
      121 acacggagtt caaattccca cagattaagg ctcgtccgtc gcacaaggta atgtgtgaat
      181 attatatctg tcgtgcaaaa ttgcctggcc tgcacaattg ctgttatagt tggcggcagg
      241 gagagtttta acattgacta gcgtgctgat aatttgtgag aaataataat tgacaagtag
      301 atactgacat ttgagaagag cttctgaact gttattagta acaaaaatgg aaagctgatg
      361 cacggaaaaa ggaaagaaaa agccatactt ttttttaggt aggaaaagaa aaagccatac
      421 gagactgatg tctctcagat gggccgggat ctgtctatct agcaggcagc agcccaccaa
      481 cctcacgggc cagcaattac gagtccttct aaaagctccc gccgaggggc gctggcgctg
      541 ctgtgcagca gcacgtctaa cattagtccc acctcgccag tttacaggga gcagaaccag
      601 cttataagcg gaggcgcggc accaagaagc gtgagaccaa cccagtggac ataagcctgt
      661 tcggttcgta agctgtaatg caagtagcgt atgcgctcac gcaactggtc cagaaccttg
      721 accgaacgca gcggtggtaa cggcgcagtg gcggttttca tggcttgtta tgactgtttt
      781 tttggggtac agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc cgtgggtcga
      841 tgtttgatgt tatggagcag caacgatgtt acgcagcagg gcagtcgccc taaaacaaag
      901 ttaaacatca tgggggaagc ggtgatcgcc gaagtatcga ctcaactatc agaggtagtt
      961 ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg tacatttgta cggctccgca
     1021 gtggatggcg gcctgaagcc acacagtgat attgatttgc tggttacggt gaccgtaagg
     1081 cttgatgaaa caacgcggcg agctttgatc aacgaccttt tggaaacttc ggcttcccct
     1141 ggagagagcg agattctccg cgctgtagaa gtcaccattg ttgtgcacga cgacatcatt
     1201 ccgtggcgtt atccagctaa gcgcgaactg caatttggag aatggcagcg caatgacatt
     1261 cttgcaggta tcttcgagcc agccacgatc gacattgatc tggctatctt gctgacaaaa
     1321 gcaagagaac atagcgttgc cttggtaggt ccagcggcgg aggaactctt tgatccggtt
     1381 cctgaacagg atctatttga ggcgctaaat gaaaccttaa cgctatggaa ctcgccgccc
     1441 gactgggctg gcgatgagcg aaatgtagtg cttacgttgt cccgcatttg gtacagcgca
     1501 gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact gggcaatgga gcgcctgccg
     1561 gcccagtatc agcccgtcat acttgaagct agacaggctt atcttggaca agaagaagat
     1621 cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc actacgtgaa aggcgagatc
     1681 accaaggtag tcggcaaata atgtctagct agaaattcgt tcaagccgac gccgcttcgc
     1741 ggcgcggctt aactcaagcg ttagatgcac taagcacata attgctcaca gccaaactat
     1801 caggtcaagt ctgcttttat tatttttaag cgtgcataat aagccggtct cggttttaga
     1861 gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga
     1921 gtcggtgctt ttttttttgt ccttctgttt ttttagtcag tctctttttt cagaagtaca
     1981 acatcttttt tttgtccttc tgttttttta gtcagtctct tttttcagaa gtacaacatc
     2041 ttttttttgt ccttctgttt ttttagtcag tctctttttt cagaagtaca acatcttttt
     2101 tttgtccttc tgttttttta gtcagtctct tttttcagaa gtactctatg atgtatcgtt
     2161 ctgggaaatg tctgtctgtc tacaacccca taatctatat ttgcaatcac acatctaata
     2221 ttctctgtga caagacagcc gaacaagctt gatatcgaat tcctgcagtg cagcgtgacc
     2281 cggtcgtgcc cctctctaga gataatgagc attgcatgtc taagttataa aaaattacca
     2341 catatttttt ttgtcacact tgtttgaagt gcagtttatc tatctttata catatattta
     2401 aactttactc tacgaataat ataatctata gtactacaat aatatcagtg ttttagagaa
     2461 tcatataaat gaacagttag acatggtcta aaggacaatt gagtattttg acaacaggac
     2521 tctacagttt tatcttttta gtgtgcatgt gttctccttt ttttttgcaa atagcttcac
     2581 ctatataata cttcatccat tttattagta catccattta gggtttaggg ttaatggttt
     2641 ttatagacta atttttttag tacatctatt ttattctatt ttagcctcta aattaagaaa
     2701 actaaaactc tattttagtt tttttattta ataatttaga tataaaatag aataaaataa
     2761 agtgactaaa aattaaacaa atacccttta agaaattaaa aaaactaagg aaacattttt
     2821 cttgtttcga gtagataatg ccagcctgtt aaacgccgtc gacgagtcta acggacacca
     2881 accagcgaac cagcagcgtc gcgtcgggcc aagcgaagca gacggcacgg catctctgtc
     2941 gctgcctctg gacccctctc gagagttccg ctccaccgtt ggacttgctc cgctgtcggc
     3001 atccagaaat gcgtggcgga gcggcagacg tgagccggca cggcaggcgg cctcctcctc
     3061 ctctcacggc acggcagcta cgggggattc ctttcccacc gctccttcgc tttcccttcc
     3121 tcgcccgccg taataaatag acaccccctc cacaccctct ttccccaacc tcgtgttgtt
     3181 cggagcgcac acacacacaa ccagatctcc cccaaatcca cccgtcggca cctccgcttc
     3241 aaggtacgcc gctcgtcctc cccccccccc cctctctacc ttctctagat cggcgttccg
     3301 gtccatggtt agggcccggt agttctactt ctgttcatgt ttgtgttaga tccgtgtttg
     3361 tgttagatcc gtgctgctag cgttcgtaca cggatgcgac ctgtacgtca gacacgttct
     3421 gattgctaac ttgccagtgt ttctctttgg ggaatcctgg gatggctcta gccgttccgc
     3481 agacgggatc gatttcatga ttttttttgt ttcgttgcat agggtttggt ttgccctttt
     3541 cctttatttc aatatatgcc gtgcacttgt ttgtcgggtc atcttttcat gctttttttt
     3601 gtcttggttg tgatgatgtg gtctggttgg gcggtcgttc tagatcggag tagaattctg
     3661 tttcaaacta cctggtggat ttattaattt tggatctgta tgtgtgtgcc atacatattc
     3721 atagttacga attgaagatg atggatggaa atatcgatct aggataggta tacatgttga
     3781 tgcgggtttt actgatgcat atacagagat gctttttgtt cgcttggttg tgatgatgtg
     3841 gtgtggttgg gcggtcgttc attcgttcta gatcggagta gaatactgtt tcaaactacc
     3901 tggtgtattt attaattttg gaactgtatg tgtgtgtcat acatcttcat agttacgagt
     3961 ttaagatgga tggaaatatc gatctaggat aggtatacat gttgatgtgg gttttactga
     4021 tgcatataca tgatggcata tgcagcatct attcatatgc tctaaccttg agtacctatc
     4081 tattataata aacaagtatg ttttataatt attttgatct tgatatactt ggatgatggc
     4141 atatgcagca gctatatgtg gattttttta gccctgcctt catacgctat ttatttgctt
     4201 ggtactgttt cttttgtcga tgctcaccct gttgtttggt gttacttctg cagcccgggg
     4261 gatccccaat acttgtatgg ccgcggccgc tctagatgga ttacaaggac cacgacgggg
     4321 attacaagga ccacgacatt gattacaagg atgatgatga caagatggct ccgaagaaga
     4381 agaggaaggt tggcatccac ggggtgccag ctgctgacaa gaagtactcg atcggcctcg
     4441 atattgggac taactctgtt ggctgggccg tgatcaccga cgagtacaag gtgccctcaa
     4501 agaagttcaa ggtcctgggc aacaccgatc ggcattccat caagaagaat ctcattggcg
     4561 ctctcctgtt cgacagcggc gagacggctg aggctacgcg gctcaagcgc accgcccgca
     4621 ggcggtacac gcgcaggaag aatcgcatct gctacctgca ggagattttc tccaacgaga
     4681 tggcgaaggt tgacgattct ttcttccaca ggctggagga gtcattcctc gtggaggagg
     4741 ataagaagca cgagcggcat ccaatcttcg gcaacattgt cgacgaggtt gcctaccacg
     4801 agaagtaccc tacgatctac catctgcgga agaagctcgt ggactccaca gataaggcgg
     4861 acctccgcct gatctacctc gctctggccc acatgattaa gttcaggggc catttcctga
     4921 tcgaggggga tctcaacccg gacaatagcg atgttgacaa gctgttcatc cagctcgtgc
     4981 agacgtacaa ccagctcttc gaggagaacc ccattaatgc gtcaggcgtc gacgcgaagg
     5041 ctatcctgtc cgctaggctc tcgaagtctc ggcgcctcga gaacctgatc gcccagctgc
     5101 cgggcgagaa gaagaacggc ctgttcggga atctcattgc gctcagcctg gggctcacgc
     5161 ccaacttcaa gtcgaatttc gatctcgctg aggacgccaa gctgcagctc tccaaggaca
     5221 catacgacga tgacctggat aacctcctgg cccagatcgg cgatcagtac gcggacctgt
     5281 tcctcgctgc caagaatctg tcggacgcca tcctcctgtc tgatattctc agggtgaaca
     5341 ccgagattac gaaggctccg ctctcagcct ccatgatcaa gcgctacgac gagcaccatc
     5401 aggatctgac cctcctgaag gcgctggtca ggcagcagct ccccgagaag tacaaggaga
     5461 tcttcttcga tcagtcgaag aacggctacg ctgggtacat tgacggcggg gcctctcagg
     5521 aggagttcta caagttcatc aagccgattc tggagaagat ggacggcacg gaggagctgc
     5581 tggtgaagct caatcgcgag gacctcctga ggaagcagcg gacattcgat aacggcagca
     5641 tcccacacca gattcatctc ggggagctgc acgctatcct gaggaggcag gaggacttct
     5701 accctttcct caaggataac cgcgagaaga tcgagaagat tctgactttc aggatcccgt
     5761 actacgtcgg cccactcgct aggggcaact cccgcttcgc ttggatgacc cgcaagtcag
     5821 aggagacgat cacgccgtgg aacttcgagg aggtggtcga caagggcgct agcgctcagt
     5881 cgttcatcga gaggatgacg aatttcgaca agaacctgcc aaatgagaag gtgctcccta
     5941 agcactcgct cctgtacgag tacttcacag tctacaacga gctgactaag gtgaagtatg
     6001 tgaccgaggg catgaggaag ccggctttcc tgtctgggga gcagaagaag gccatcgtgg
     6061 acctcctgtt caagaccaac cggaaggtca cggttaagca gctcaaggag gactacttca
     6121 agaagattga gtgcttcgat tcggtcgaga tctctggcgt tgaggaccgc ttcaacgcct
     6181 ccctggggac ctaccacgat ctcctgaaga tcattaagga taaggacttc ctggacaacg
     6241 aggagaatga ggatatcctc gaggacattg tgctgacact cactctgttc gaggaccggg
     6301 agatgatcga ggagcgcctg aagacttacg cccatctctt cgatgacaag gtcatgaagc
     6361 agctcaagag gaggaggtac accggctggg ggaggctgag caggaagctc atcaacggca
     6421 ttcgggacaa gcagtccggg aagacgatcc tcgacttcct gaagagcgat ggcttcgcga
     6481 accgcaattt catgcagctg attcacgatg acagcctcac attcaaggag gatatccaga
     6541 aggctcaggt gagcggccag ggggactcgc tgcacgagca tatcgcgaac ctcgctggct
     6601 cgccagctat caagaagggg attctgcaga ccgtgaaggt tgtggacgag ctggtgaagg
     6661 tcatgggcag gcacaagcct gagaacatcg tcattgagat ggcccgggag aatcagacca
     6721 cgcagaaggg ccagaagaac tcacgcgaga ggatgaagag gatcgaggag ggcattaagg
     6781 agctggggtc ccagatcctc aaggagcacc cggtggagaa cacgcagctg cagaatgaga
     6841 agctctacct gtactacctc cagaatggcc gcgatatgta tgtggaccag gagctggata
     6901 ttaacaggct cagcgattac gacgtcgatc atatcgttcc acagtcattc ctgaaggatg
     6961 actccattga caacaaggtc ctcaccaggt cggacaagaa ccggggcaag tctgataatg
     7021 ttccttcaga ggaggtcgtt aagaagatga agaactactg gcgccagctc ctgaatgcca
     7081 agctgatcac gcagcggaag ttcgataacc tcacaaaggc tgagaggggc gggctctctg
     7141 agctggacaa ggcgggcttc atcaagaggc agctggtcga gacacggcag atcactaagc
     7201 acgttgcgca gattctcgac tcacggatga acactaagta cgatgagaat gacaagctga
     7261 tccgcgaggt gaaggtcatc accctgaagt caaagctcgt ctccgacttc aggaaggatt
     7321 tccagttcta caaggttcgg gagatcaaca attaccacca tgcccatgac gcgtacctga
     7381 acgcggtggt cggcacagct ctgatcaaga agtacccaaa gctcgagagc gagttcgtgt
     7441 acggggacta caaggtttac gatgtgagga agatgatcgc caagtcggag caggagattg
     7501 gcaaggctac cgccaagtac ttcttctact ctaacattat gaatttcttc aagacagaga
     7561 tcactctggc caatggcgag atccggaagc gccccctcat cgagacgaac ggcgagacgg
     7621 gggagatcgt gtgggacaag ggcagggatt tcgcgaccgt caggaaggtt ctctccatgc
     7681 cacaagtgaa tatcgtcaag aagacagagg tccagactgg cgggttctct aaggagtcaa
     7741 ttctgcctaa gcggaacagc gacaagctca tcgcccgcaa gaaggactgg gatccgaaga
     7801 agtacggcgg gttcgacagc cccactgtgg cctactcggt cctggttgtg gcgaaggttg
     7861 agaagggcaa gtccaagaag ctcaagagcg tgaaggagct gctggggatc acgattatgg
     7921 agcgctccag cttcgagaag aacccgatcg atttcctgga ggcgaagggc tacaaggagg
     7981 tgaagaagga cctgatcatt aagctcccca agtactcact cttcgagctg gagaacggca
     8041 ggaagcggat gctggcttcc gctggcgagc tgcagaaggg gaacgagctg gctctgccgt
     8101 ccaagtatgt gaacttcctc tacctggcct cccactacga gaagctcaag ggcagccccg
     8161 aggacaacga gcagaagcag ctgttcgtcg agcagcacaa gcattacctc gacgagatca
     8221 ttgagcagat ttccgagttc tccaagcgcg tgatcctggc cgacgcgaat ctggataagg
     8281 tcctctccgc gtacaacaag caccgcgaca agccaatcag ggagcaggct gagaatatca
     8341 ttcatctctt caccctgacg aacctcggcg cccctgctgc tttcaagtac ttcgacacaa
     8401 ctatcgatcg caagaggtac acaagcacta aggaggtcct ggacgcgacc ctcatccacc
     8461 agtcgattac cggcctctac gagacgcgca tcgacctgtc tcagctcggg ggcgacaagc
     8521 ggccagcggc gacgaagaag gcggggcagg cgaagaagaa gaagtgagct caggcctcca
     8581 tacaagtatt ggggatccga atttccccga tcgttcaaac atttggcaat aaagtttctt
     8641 aagattgaat cctgttgccg gtcttgcgat gattatcata taatttctgt tgaattacgt
     8701 taagcatgta ataattaaca tgtaatgcat gacgttattt atgagatggg tttttatgat
     8761 tagagtcccg caattataca tttaatacgc gatagaaaac aaaatatagc gcgcaaacta
     8821 ggataaatta tcgcgcgcgg tgtcatctat gttactagat cgggaattca ctggccgtcg
     8881 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     8941 atcccccttt cgccaggggt accgagctct agaattcgag gcggtttgcg tattggctag
     9001 agcagcttgc caacatggtg gagcacgaca ctctcgtcta ctccaagaat atcaaagata
     9061 cagtctcaga agaccaaagg gctattgaga cttttcaaca aagggtaata tcgggaaacc
     9121 tcctcggatt ccattgccca gctatctgtc acttcatcaa aaggacagta gaaaaggaag
     9181 gtggcaccta caaatgccat cattgcgata aaggaaaggc tatcgttcaa gatgcctctg
     9241 ccgacagtgg tcccaaagat ggacccccac ccacgaggag catcgtggaa aaagaagacg
     9301 ttccaaccac gtcttcaaag caagtggatt gatgtgataa catggtggag cacgacactc
     9361 tcgtctactc caagaatatc aaagatacag tctcagaaga ccaaagggct attgagactt
     9421 ttcaacaaag ggtaatatcg ggaaacctcc tcggattcca ttgcccagct atctgtcact
     9481 tcatcaaaag gacagtagaa aaggaaggtg gcacctacaa atgccatcat tgcgataaag
     9541 gaaaggctat cgttcaagat gcctctgccg acagtggtcc caaagatgga cccccaccca
     9601 cgaggagcat cgtggaaaaa gaagacgttc caaccacgtc ttcaaagcaa gtggattgat
     9661 gtgatatctc cactgacgta agggatgacg cacaatccca ctatccttcg caagaccttc
     9721 ctctatataa ggaagttcat ttcatttgga gaggacacgc tgaaatcacc agtctctctc
     9781 tacaaatcta tctctctcga gtctaccatg agcccagaac gacgcccggc cgacatccgc
     9841 cgtgccaccg aggcggacat gccggcggtc tgcaccatcg tcaaccacta catcgagaca
     9901 agcacggtca acttccgtac cgagccgcag gaaccgcagg agtggacgga cgacctcgtc
     9961 cgtctgcggg agcgctatcc ctggctcgtc gccgaggtgg acggcgaggt cgccggcatc
    10021 gcctacgcgg gcccctggaa ggcacgcaac gcctacgact ggacggccga gtcgaccgtg
    10081 tacgtctccc cccgccacca gcggacggga ctgggctcca cgctctacac ccacctgctg
    10141 aagtccctgg aggcacaggg cttcaagagc gtggtcgctg tcatcgggct gcccaacgac
    10201 ccgagcgtgc gcatgcacga ggcgctcgga tatgcccccc gcggcatgct gcgggcggcc
    10261 ggcttcaagc acgggaactg gcatgacgtg ggtttctggc agctggactt cagcctgccg
    10321 gtaccgcccc gtccggtcct gcccgtcacc gagatttgac tcgagtttct ccataataat
    10381 gtgtgagtag ttcccagata agggaattag ggttcctata gggtttcgct catgtgttga
    10441 gcatataaga aacccttagt atgtatttgt atttgtaaaa tacttctatc aataaaattt
    10501 ctaattccta aaaccaaaat ccagtactaa aatccagatc ccccgaatta attcggcgtt
    10561 aattcagtac attaaaaacg tccgcaatgt gttattaagt tgtcactagt caggttaact
    10621 caattcggcg ttaattcagt acattaaaaa cgtccgcaat gtgttattaa gttgtctaag
    10681 cgtcaatttg tttacaccac aatatatcct gccaccagcc agccaacagc tccccgaccg
    10741 gcagctcggc acaaaatcac cactcgatac aggcagccca tcagtccggg acggcgtcag
    10801 cgggagagcc gttgtaaggc ggcagacttt gctcatgtta ccgatgctat tcggaagaac
    10861 ggcaactaag ctgccgggtt tgaaacacgg atgatctcgc ggagggtagc atgttgattg
    10921 taacgatgac agagcgttgc tgcctgtgat cacttaagta actaactaac aggaagagtt
    10981 tgtagaaacg caaaaaggcc atccgtcagg atggccttct gcttagtttg atgcctggca
    11041 gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac gttcaaatcc
    11101 gctcccggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga
    11161 aaggcccagt cttccgactg agcctttcgt tttatttgat gcctggcagt tccctactct
    11221 cgcttagtag ttagacgtcc ccgagatcca tgctagacca tgaatccaga agcccgagag
    11281 gttgccgcct ttcgggcttt ttctttttca aaaaaaaaaa tttataaaac gatctgttgc
    11341 ggccggccgc cgggttgtgg gcaaaggcgc tcgacggtgg gcaaccgctt gcggttgtcc
    11401 acgggcggag ccggtgcgcg tagcgcattg tccacaagcc aagggcgacc aataattgat
    11461 atatatattc ataattgaaa agctaattga acatactact tgctgtaact acttgccgga
    11521 gcgaggggtg tttgcaagct gttgatctga aagggctatt agcgttctca cgtgcctttt
    11581 tgattagcga tttcacgtga ccttattagc gatttcacgt actccgatta gcgatttcac
    11641 gtaccctgat tagcgatttc acgtggatag tttttggagc gggccggaaa gccccgtgaa
    11701 tcaaggcttt gcggggcatt agcggtttca cgtggataac taccctctat ccacaggctt
    11761 ccggggataa aaaagcccgc tcgacggcgg gctgttggat ggggatctag cggtaatacg
    11821 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
    11881 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
    11941 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
    12001 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
    12061 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
    12121 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
    12181 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
    12241 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
    12301 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
    12361 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
    12421 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
    12481 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctaccg ggtctgacgc
    12541 tcagtggaac ggggcccaat ctgaataatg ttacaaccaa ttaaccaatt ctgattagaa
    12601 aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata
    12661 tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat
    12721 ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa
    12781 tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc
    12841 cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt
    12901 acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg
    12961 agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa
    13021 ccggcgcagg gacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc
    13081 taatacctgg aatgctgttt ttccggggat cgcagtggtg agtaaccatg catcatcagg
    13141 agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct
    13201 gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc
    13261 tggcgcatcg ggcttcccat acaagcgata gattgtcgca cctgattgcc cgacattatc
    13321 gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctcga
    13381 cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag
    13441 ttttattgtt catgatgata tatttttatc ttgtgcaatg taacatcaga gattttgaga
    13501 cacgggccag agctgcagtt tgatcccgag gggaaccctg tggttgacat gcacatacaa
    13561 atggacgaac ggataaacct tttcacgccc ttttaaatat ccgttattct aataaacgct
    13621 cttttctctt ag
//
