LOCUS       sequence-259001-       16713 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pGreen3 CRISPR/Cas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16713)
  AUTHORS   Zhang Q, Zhang Y, Lu MH, Chai YP, Jiang YY, Zhou Y, Wang XC, Chen QJ
  TITLE     A Novel Ternary Vector System United with Morphogenic Genes Enhances
            CRISPR/Cas Delivery in Maize.
  JOURNAL   Plant Physiol. 2019 Sep 26. pii: pp.19.00767. doi: 
            10.1104/pp.19.00767.
  PUBMED    31558579
REFERENCE   2  (bases 1 to 16713)
  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 
            Physiol. 2019 Sep 26. pii: pp.19.00767. doi: 10.1104/pp.19.00767."
COMMENT     Sequence Label: pG3GB411
FEATURES             Location/Qualifiers
     source          1..16713
                     /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"
     primer_bind     213..235
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     227..244
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     228..244
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        311..690
                     /label=OsU3 promoter
                     /note="Oryza sativa (rice) snRNA U3 promoter"
     promoter        713..817
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(818..886,887..1678)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1036..1055)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     misc_RNA        1827..1902
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        1968..3960
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     CDS             3979..4044
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4051..4071
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4096..8196
                     /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             4096..8196
                     /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             8109..8117
                     /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             8197..8244
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      11517..11769
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(11794..11810)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11794..11810)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11807..11829)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11818..11834
                     /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)."
     promoter        complement(join(11842..11848,11849..11866,11867..11872))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11887..11908
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        12099..12775
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     12712..12732
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             12820..13371
                     /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    13378..13552
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    13630..13654
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     complement(14032..14051)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             14043..14852
                     /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     complement(14110..14129)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      14931..15017
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      15109..15136
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      15215..15803
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     15704..15723
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     rep_origin      15918..16528
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     rep_origin      16005..16536
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 gtttacccgc caatatatcc tgtcaaacac tgatagttta aactgaaggc gggaaacgac
       61 aatctgatcc aagctcaagc tgctctagca ttcgccattc aggctgcgca actgttggga
      121 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
      181 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
      241 cagtgccaag cttagtaatt catccaggtc accaagttct aggattttca gaactgcaac
      301 ttattttatc aaggaatctt taaacatacg aacagatcac ttaaagttct tctgaagcaa
      361 cttaaagtta tcaggcttgc atggatcttg gaggaatcag atgtgcagtc agggaccata
      421 gcacaagaca ggcgtcttct actggtgcta ccagcaaatg ctggaagccg ggaacactgg
      481 gtacgttgga aaccacgtga tgtgaagaag taagataaac tgtaggagaa aagcatttcg
      541 tagtgggcca tgaagccttt caggacatgt attgcagtat gggccggccc attacgcaat
      601 tggacgacaa caaagtctag tattagtacc acctcggcta tccacataga tcaaagctga
      661 tttaaaagag ttgtgcagat gatccgtggc gtgagacctt cggggaaatg tgcgcggaac
      721 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatgg gacaataacc
      781 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
      841 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
      901 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
      961 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     1021 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     1081 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     1141 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     1201 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     1261 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     1321 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     1381 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     1441 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     1501 tattgctgat aaatctggag ccggtgagcg tggctctcgc ggtatcattg cagcactggg
     1561 gccagatggt aagccctccc gtatcgtagt tatctacacc acggggagtc aggcaactat
     1621 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     1681 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     1741 aaggatctag gtgaagatcc tttttgataa tcccatgacc aaaatccctt aacgtgagtt
     1801 ttcgttccac tgagcgtcag gtctcagttt tagagctaga aatagcaagt taaaataagg
     1861 ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gctttttttt ttgtccttct
     1921 gtttttttag tcagtctctt ttttcagaag tacaacatct tgcatgcctg cagtgcagcg
     1981 tgacccggtc gtgcccctct ctagagataa tgagcattgc atgtctaagt tataaaaaat
     2041 taccacatat tttttttgtc acacttgttt gaagtgcagt ttatctatct ttatacatat
     2101 atttaaactt tactctacga ataatataat ctatagtact acaataatat cagtgtttta
     2161 gagaatcata taaatgaaca gttagacatg gtctaaagga caattgagta ttttgacaac
     2221 aggactctac agttttatct ttttagtgtg catgtgttct cctttttttt tgcaaatagc
     2281 ttcacctata taatacttca tccattttat tagtacatcc atttagggtt tagggttaat
     2341 ggtttttata gactaatttt tttagtacat ctattttatt ctattttagc ctctaaatta
     2401 agaaaactaa aactctattt tagttttttt atttaataat ttagatataa aatagaataa
     2461 aataaagtga ctaaaaatta aacaaatacc ctttaagaaa ttaaaaaaac taaggaaaca
     2521 tttttcttgt ttcgagtaga taatgccagc ctgttaaacg ccgtcgacga gtctaacgga
     2581 caccaaccag cgaaccagca gcgtcgcgtc gggccaagcg aagcagacgg cacggcatct
     2641 ctgtcgctgc ctctggaccc ctctcgagag ttccgctcca ccgttggact tgctccgctg
     2701 tcggcatcca gaaattgcgt ggcggagcgg cagacgtgag ccggcacggc aggcggcctc
     2761 ctcctcctct cacggcaccg gcagctacgg gggattcctt tcccaccgct ccttcgcttt
     2821 cccttcctcg cccgccgtaa taaatagaca ccccctccac accctctttc cccaacctcg
     2881 tgttgttcgg agcgcacaca cacacaacca gatctccccc aaatccaccc gtcggcacct
     2941 ccgcttcaag gtacgccgct cgtcctcccc ccccccccct ctctaccttc tctagatcgg
     3001 cgttccggtc catggttagg gcccggtagt tctacttctg ttcatgtttg tgttagatcc
     3061 gtgtttgtgt tagatccgtg ctgctagcgt tcgtacacgg atgcgacctg tacgtcagac
     3121 acgttctgat tgctaacttg ccagtgtttc tctttgggga atcctgggat ggctctagcc
     3181 gttccgcaga cgggatcgat ttcatgattt tttttgtttc gttgcatagg gtttggtttg
     3241 cccttttcct ttatttcaat atatgccgtg cacttgtttg tcgggtcatc ttttcatgct
     3301 tttttttgtc ttggttgtga tgatgtggtc tggttgggcg gtcgttctag atcggagtag
     3361 aattctgttt caaactacct ggtggattta ttaattttgg atctgtatgt gtgtgccata
     3421 catattcata gttacgaatt gaagatgatg gatggaaata tcgatctagg ataggtatac
     3481 atgttgatgc gggttttact gatgcatata cagagatgct ttttgttcgc ttggttgtga
     3541 tgatgtggtg tggttgggcg gtcgttcatt cgttctagat cggagtagaa tactgtttca
     3601 aactacctgg tgtatttatt aattttggaa ctgtatgtgt gtgtcataca tcttcatagt
     3661 tacgagttta agatggatgg aaatatcgat ctaggatagg tatacatgtt gatgtgggtt
     3721 ttactgatgc atatacatga tggcatatgc agcatctatt catatgctct aaccttgagt
     3781 acctatctat tataataaac aagtatgttt tataattatt ttgatcttga tatacttgga
     3841 tgatggcata tgcagcagct atatgtggat ttttttagcc ctgccttcat acgctattta
     3901 tttgcttggt actgtttctt ttgtcgatgc tcaccctgtt gtttggtgtt acttctgcag
     3961 ccctaggcct actagatgga ttacaaggac cacgacgggg attacaagga ccacgacatt
     4021 gattacaagg atgatgatga caagatggct ccgaagaaga agaggaaggt tggcatccac
     4081 ggggtgccag ctgctgacaa gaagtactcg atcggcctcg atattgggac taactctgtt
     4141 ggctgggccg tgatcaccga cgagtacaag gtgccctcaa agaagttcaa ggtcctgggc
     4201 aacaccgatc ggcattccat caagaagaat ctcattggcg ctctcctgtt cgacagcggc
     4261 gagacggctg aggctacgcg gctcaagcgc accgcccgca ggcggtacac gcgcaggaag
     4321 aatcgcatct gctacctgca ggagattttc tccaacgaga tggcgaaggt tgacgattct
     4381 ttcttccaca ggctggagga gtcattcctc gtggaggagg ataagaagca cgagcggcat
     4441 ccaatcttcg gcaacattgt cgacgaggtt gcctaccacg agaagtaccc tacgatctac
     4501 catctgcgga agaagctcgt ggactccaca gataaggcgg acctccgcct gatctacctc
     4561 gctctggccc acatgattaa gttcaggggc catttcctga tcgaggggga tctcaacccg
     4621 gacaatagcg atgttgacaa gctgttcatc cagctcgtgc agacgtacaa ccagctcttc
     4681 gaggagaacc ccattaatgc gtcaggcgtc gacgcgaagg ctatcctgtc cgctaggctc
     4741 tcgaagtctc ggcgcctcga gaacctgatc gcccagctgc cgggcgagaa gaagaacggc
     4801 ctgttcggga atctcattgc gctcagcctg gggctcacgc ccaacttcaa gtcgaatttc
     4861 gatctcgctg aggacgccaa gctgcagctc tccaaggaca catacgacga tgacctggat
     4921 aacctcctgg cccagatcgg cgatcagtac gcggacctgt tcctcgctgc caagaatctg
     4981 tcggacgcca tcctcctgtc tgatattctc agggtgaaca ccgagattac gaaggctccg
     5041 ctctcagcct ccatgatcaa gcgctacgac gagcaccatc aggatctgac cctcctgaag
     5101 gcgctggtca ggcagcagct ccccgagaag tacaaggaga tcttcttcga tcagtcgaag
     5161 aacggctacg ctgggtacat tgacggcggg gcctctcagg aggagttcta caagttcatc
     5221 aagccgattc tggagaagat ggacggcacg gaggagctgc tggtgaagct caatcgcgag
     5281 gacctcctga ggaagcagcg gacattcgat aacggcagca tcccacacca gattcatctc
     5341 ggggagctgc acgctatcct gaggaggcag gaggacttct accctttcct caaggataac
     5401 cgcgagaaga tcgagaagat tctgactttc aggatcccgt actacgtcgg cccactcgct
     5461 aggggcaact cccgcttcgc ttggatgacc cgcaagtcag aggagacgat cacgccgtgg
     5521 aacttcgagg aggtggtcga caagggcgct agcgctcagt cgttcatcga gaggatgacg
     5581 aatttcgaca agaacctgcc aaatgagaag gtgctcccta agcactcgct cctgtacgag
     5641 tacttcacag tctacaacga gctgactaag gtgaagtatg tgaccgaggg catgaggaag
     5701 ccggctttcc tgtctgggga gcagaagaag gccatcgtgg acctcctgtt caagaccaac
     5761 cggaaggtca cggttaagca gctcaaggag gactacttca agaagattga gtgcttcgat
     5821 tcggtcgaga tctctggcgt tgaggaccgc ttcaacgcct ccctggggac ctaccacgat
     5881 ctcctgaaga tcattaagga taaggacttc ctggacaacg aggagaatga ggatatcctc
     5941 gaggacattg tgctgacact cactctgttc gaggaccggg agatgatcga ggagcgcctg
     6001 aagacttacg cccatctctt cgatgacaag gtcatgaagc agctcaagag gaggaggtac
     6061 accggctggg ggaggctgag caggaagctc atcaacggca ttcgggacaa gcagtccggg
     6121 aagacgatcc tcgacttcct gaagagcgat ggcttcgcga accgcaattt catgcagctg
     6181 attcacgatg acagcctcac attcaaggag gatatccaga aggctcaggt gagcggccag
     6241 ggggactcgc tgcacgagca tatcgcgaac ctcgctggct cgccagctat caagaagggg
     6301 attctgcaga ccgtgaaggt tgtggacgag ctggtgaagg tcatgggcag gcacaagcct
     6361 gagaacatcg tcattgagat ggcccgggag aatcagacca cgcagaaggg ccagaagaac
     6421 tcacgcgaga ggatgaagag gatcgaggag ggcattaagg agctggggtc ccagatcctc
     6481 aaggagcacc cggtggagaa cacgcagctg cagaatgaga agctctacct gtactacctc
     6541 cagaatggcc gcgatatgta tgtggaccag gagctggata ttaacaggct cagcgattac
     6601 gacgtcgatc atatcgttcc acagtcattc ctgaaggatg actccattga caacaaggtc
     6661 ctcaccaggt cggacaagaa ccggggcaag tctgataatg ttccttcaga ggaggtcgtt
     6721 aagaagatga agaactactg gcgccagctc ctgaatgcca agctgatcac gcagcggaag
     6781 ttcgataacc tcacaaaggc tgagaggggc gggctctctg agctggacaa ggcgggcttc
     6841 atcaagaggc agctggtcga gacacggcag atcactaagc acgttgcgca gattctcgac
     6901 tcacggatga acactaagta cgatgagaat gacaagctga tccgcgaggt gaaggtcatc
     6961 accctgaagt caaagctcgt ctccgacttc aggaaggatt tccagttcta caaggttcgg
     7021 gagatcaaca attaccacca tgcccatgac gcgtacctga acgcggtggt cggcacagct
     7081 ctgatcaaga agtacccaaa gctcgagagc gagttcgtgt acggggacta caaggtttac
     7141 gatgtgagga agatgatcgc caagtcggag caggagattg gcaaggctac cgccaagtac
     7201 ttcttctact ctaacattat gaatttcttc aagacagaga tcactctggc caatggcgag
     7261 atccggaagc gccccctcat cgagacgaac ggcgagacgg gggagatcgt gtgggacaag
     7321 ggcagggatt tcgcgaccgt caggaaggtt ctctccatgc cacaagtgaa tatcgtcaag
     7381 aagacagagg tccagactgg cgggttctct aaggagtcaa ttctgcctaa gcggaacagc
     7441 gacaagctca tcgcccgcaa gaaggactgg gatccgaaga agtacggcgg gttcgacagc
     7501 cccactgtgg cctactcggt cctggttgtg gcgaaggttg agaagggcaa gtccaagaag
     7561 ctcaagagcg tgaaggagct gctggggatc acgattatgg agcgctccag cttcgagaag
     7621 aacccgatcg atttcctgga ggcgaagggc tacaaggagg tgaagaagga cctgatcatt
     7681 aagctcccca agtactcact cttcgagctg gagaacggca ggaagcggat gctggcttcc
     7741 gctggcgagc tgcagaaggg gaacgagctg gctctgccgt ccaagtatgt gaacttcctc
     7801 tacctggcct cccactacga gaagctcaag ggcagccccg aggacaacga gcagaagcag
     7861 ctgttcgtcg agcagcacaa gcattacctc gacgagatca ttgagcagat ttccgagttc
     7921 tccaagcgcg tgatcctggc cgacgcgaat ctggataagg tcctctccgc gtacaacaag
     7981 caccgcgaca agccaatcag ggagcaggct gagaatatca ttcatctctt caccctgacg
     8041 aacctcggcg cccctgctgc tttcaagtac ttcgacacaa ctatcgatcg caagaggtac
     8101 acaagcacta aggaggtcct ggacgcgacc ctcatccacc agtcgattac cggcctctac
     8161 gagacgcgca tcgacctgtc tcagctcggg ggcgacaagc ggccagcggc gacgaagaag
     8221 gcggggcagg cgaagaagaa gaagtgagct cagagctttc gttcgtatca tcggtttcga
     8281 caacgttcgt caagttcaat gcatcagttt cattgcgcac acaccagaat cctactgagt
     8341 ttgagtatta tggcattggg aaaactgttt ttcttgtacc atttgttgtg cttgtaattt
     8401 actgtgtttt ttattcggtt ttcgctatcg aactgtgaaa tggaaatgga tggagaagag
     8461 ttaatgaatg atatggtcct tttgttcatt ctcaaattaa tattatttgt tttttctctt
     8521 atttgttgtg tgttgaattt gaaattataa gagatatgca aacattttgt tttgagtaaa
     8581 aatgtgtcaa atcgtggcct ctaatgaccg aagttaatat gaggagtaaa acacttgtag
     8641 ttgtaccatt atgcttattc actaggcaac aaatatattt tcagacctag aaaagctgca
     8701 aatgttactg aatacaagta tgtcctcttg tgttttagac atttatgaac tttcctttat
     8761 gtaattttcc agaatccttg tcagattcta atcattgctt tataattata gttatactca
     8821 tggatttgta gttgagtatg aaaatatttt ttaatgcatt ttatgacttg ccaattgatt
     8881 gacaacgaat tcggatcctc catgcctaca tcaactaatt tgattccttg agtttacgtt
     8941 tagtgatatg tctattttta gagcttgttg gggcttcggc ctcagctcta gccagccaaa
     9001 catgttctac caagtaccct atgttggcat gatatagtga tgcattataa caataaatga
     9061 gcgagggatt gctggctgaa aaagctatac tagctgcatt tggttatagt taaccgaact
     9121 attaattgcg tgtacaacaa aataaaaaaa atgcatgttg cacattcttt cattaacatt
     9181 atgttttggt agtgtgaatt agaaatttga ttgacagtag atcgacaaac atagtttcaa
     9241 tatgcttaag ttagttatga ctttaacata tcagtctcct tgatattttc gttttagatt
     9301 cgtctctctt ctagtgtgta tgtccacctt ccatagcagt gagggttcca ttccatccct
     9361 gtaaaaaaaa atcaaccact actatttatt tctaaaagca aaatgataaa atatcatttt
     9421 tttaatagaa taagaaagta ttgggtacat agtgatgtgc acgtaggagt aaacataaaa
     9481 aaggcagatt ttctaggttg gcccaggtgt gcagtgcagc aaattattcc gcgttggcac
     9541 ggccgcagcg cagagacagc ctcaagctct caactttcca gtgaggccgg cgcctcttta
     9601 catctcattc ccccaatttc ccaccaccat cgccattgcc accacctctc ctatatctcg
     9661 ccctccccct cctccctccc acgccattcg cctccttctt gctgcagccg ccatccccgg
     9721 ttcggttctc tcctcttctt taggtgagca actgcctctc catgtccagg ccctcccggc
     9781 agggaagact cgtgtttttg taggatgaat gtttagttca agttacattt ttctgtatta
     9841 atctatagtc tttgtaaaca ctgttttgaa tgatttattt tgtgttatgc agatcagtta
     9901 ctagatggcg caagttagca gaatctgcaa tggtgtgcag aacccatctc ttatctccaa
     9961 tctctcgaaa tccagtcaac gcaaatctcc cttatcggtt tctctgaaga cgcagcagca
    10021 tccacgagct tatccgattt cgtcgtcgtg gggattgaag aagagtggga tgacgttaat
    10081 tggctctgag cttcgtcctc ttaaggtcat gtcttctgtt tccacggcgt gcatgtcgca
    10141 cggtgcaagc agccggcccg caaccgcccg caaatcctct ggcctttccg gaaccgtccg
    10201 cattcccggc gacaagtcga tctcccaccg gtccttcatg ttcggcggcc tcgcgagcgg
    10261 tgaaacgcgc atcaccggcc ttctggaagg cgaggacgtc atcaatacgg gcaaggcgat
    10321 gcaggccatg ggcgccagga tccgtaagga aggcgacacc tggatcatcg atggcgtcgg
    10381 caatggcggc ctcctggcgc ctgaggcgcc gctcgatttc ggcaatgccg ccacgggctg
    10441 ccgcctgacc atgggcctcg tcggggtcta cgatttcgac agcaccttca tcggcgacgc
    10501 ctcgctcaca aagcgcccga tgggccgcgt gttgaacccg ctgcgcgaaa tgggcgtgca
    10561 ggtcaaatcg gaagacggtg accgtcttcc cgttaccttg cgcgggccga agacgccgac
    10621 gccgatcacc taccgcgtgc cgatggcctc cgcacaggtg aagtccgccg tgctgctcgc
    10681 cggcctcaac acgcccggca tcacgacggt catcgagccg atcatgacgc gcgatcatac
    10741 ggaaaagatg ctgcagggct ttggcgccaa ccttaccgtc gagacggatg cggacggcgt
    10801 gcgcaccatc cgcctggaag gtcgcggcaa gctcaccggc caagtcatcg acgtgccggg
    10861 cgacccgtcc tcgacggcct tcccgctggt tgcggccctg cttgttccgg gctccgacgt
    10921 caccatcctc aacgtgctga tgaaccccac ccgcaccggc ctcatcctga cgctgcagga
    10981 aatgggcgcc gacatcgaag tcatcaaccc gcgccttgcc ggcggcgaag acgtggcgga
    11041 cctgcgcgtt cgctcctcca cgctgaaggg cgtcacggtg ccggaagacc gcgcgccttc
    11101 gatgatcgac gaatatccga ttctcgctgt cgccgccgcc ttcgcggaag gggcgaccgt
    11161 gatgaacggt ctggaagaac tccgcgtcaa ggaaagcgac cgcctctcgg ccgtcgccaa
    11221 tggcctcaag ctcaatggcg tggattgcga tgagggcgag acgtcgctcg tcgtgcgcgg
    11281 ccgccctgac ggcaaggggc tcggcaacgc ctcgggcgcc gccgtcgcca cccatctcga
    11341 tcaccgcatc gccatgagct tcctcgtcat gggcctcgtg tcggaaaacc ctgtcacggt
    11401 ggacgatgcc acgatgatcg ccacgagctt cccggagttc atggacctga tggccgggct
    11461 gggcgcgaag atcgaactct ccgatacgaa ggctgcctga ctcgacgaat ttccccgatc
    11521 gttcaaacat ttggcaataa agtttcttaa gattgaatcc tgttgccggt cttgcgatga
    11581 ttatcatata atttctgttg aattacgtta agcatgtaat aattaacatg taatgcatga
    11641 cgttatttat gagatgggtt tttatgatta gagtcccgca attatacatt taatacgcga
    11701 tagaaaacaa aatatagcgc gcaaactagg ataaattatc gcgcgcggtg tcatctatgt
    11761 tactagatcg ggaactaggt aattcgtaat catgtcatag ctgtttcctg tgtgaaattg
    11821 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
    11881 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
    11941 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
    12001 gcgtattggc tagagcagct tgccaacatg gtggagcacg acactctcgt ctactccaag
    12061 aatatcaaag atacagtctc agaagaccaa agggctattg agacttttca acaaagggta
    12121 atatcgggaa acctcctcgg attccattgc ccagctatct gtcacttcat caaaaggaca
    12181 gtagaaaagg aaggtggcac ctacaaatgc catcattgcg ataaaggaaa ggctatcgtt
    12241 caagatgcct ctgccgacag tggtcccaaa gatggacccc cacccacgag gagcatcgtg
    12301 gaaaaagaag acgttccaac cacgtcttca aagcaagtgg attgatgtga acatggtgga
    12361 gcacgacact ctcgtctact ccaagaatat caaagataca gtctcagaag accaaagggc
    12421 tattgagact tttcaacaaa gggtaatatc gggaaacctc ctcggattcc attgcccagc
    12481 tatctgtcac ttcatcaaaa ggacagtaga aaaggaaggt ggcacctaca aatgccatca
    12541 ttgcgataaa ggaaaggcta tcgttcaaga tgcctctgcc gacagtggtc ccaaagatgg
    12601 acccccaccc acgaggagca tcgtggaaaa agaagacgtt ccaaccacgt cttcaaagca
    12661 agtggattga tgtgatatct ccactgacgt aagggatgac gcacaatccc actatccttc
    12721 gcaagaccct tcctctatat aaggaagttc atttcatttg gagaggacac gctgaaatca
    12781 ccagtctctc tctacaaatc tatctctctc gagtctacca tgagcccaga acgacgcccg
    12841 gccgacatcc gccgtgccac cgaggcggac atgccggcgg tctgcaccat cgtcaaccac
    12901 tacatcgaga caagcacggt caacttccgt accgagccgc aggaaccgca ggagtggacg
    12961 gacgacctcg tccgtctgcg ggagcgctat ccctggctcg tcgccgaggt ggacggcgag
    13021 gtcgccggca tcgcctacgc gggcccctgg aaggcacgca acgcctacga ctggacggcc
    13081 gagtcgaccg tgtacgtctc cccccgccac cagcggacgg gactgggctc cacgctctac
    13141 acccacctcc tgaagtccct ggaggcacag ggcttcaaga gcgtggtcgc tgtcatcggg
    13201 ctgcccaacg acccgagcgt gcgcatgcac gaggcgctcg gatatgcccc acgcggcatg
    13261 ctgcgggcgg ccggcttcaa gcacgggaac tggcatgacg tgggtttctg gcagctggac
    13321 ttcagcctgc cggtaccgcc ccgtccggtc ctgcccgtca ccgagatttg actcgagttt
    13381 ctccataata atgtgtgagt agttcccaga taagggaatt agggttccta tagggtttcg
    13441 ctcatgtgtt gagcatataa gaaaccctta gtatgtattt gtatttgtaa aatacttcta
    13501 tcaataaaat ttctaattcc taaaaccaaa atccagtact aaaatccaga tcccccgaat
    13561 taattcggcg ttaattcagt acattaaaaa cgtccgcaat gtgttattaa gttgtctaag
    13621 cgtcaatttg tttacaccac aatatatcct gccaccagcc agccaacagc tccccgaccg
    13681 gcagctcggc acaaaatcac cactcgatac aggcagccca tcagtccggg acggcgtcag
    13741 cgggagagcc gttgtaaggc ggcagacttt gctcatgtta ccgatgctat tcggaagaac
    13801 ggcaactaag ctgccgggtt tgaaacacgg atgatctcgc ggagggtagc atgttgattg
    13861 taacgatgac agagcgttgc tgcctgtgat caccgcggct ggcaaacagc tattatgggt
    13921 attatgggtc tagtctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata
    13981 aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg
    14041 ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg
    14101 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
    14161 gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
    14221 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc
    14281 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
    14341 ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
    14401 atgcgctggc agtgtccctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
    14461 acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg
    14521 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
    14581 tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
    14641 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
    14701 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
    14761 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
    14821 agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacatt
    14881 attcagattg gcgtctaact actaagcgag agtagggaac tgccaggcat caaataaaac
    14941 gaaaggctca gtcggaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
    15001 tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgtgaagcaa cggcccggag
    15061 ggtggcgggc aggacgcccg ccataaactg ccaggcatca aactaagcag aaggccatcc
    15121 tgacggatgg cctttttgcg tttctacaaa ctcttcctgt tagggccccg ttccactgag
    15181 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
    15241 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
    15301 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
    15361 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
    15421 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
    15481 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
    15541 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
    15601 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
    15661 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
    15721 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
    15781 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
    15841 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
    15901 gtattaccgc tagatagctg gttgccctcg ccgctgggct ggcggccgtc tatggccctg
    15961 caaacgcgcc agaaacgccg tcgaagccgt gtgcgagaca ccccggccgc cggcgttgtg
    16021 gatacctcgc ggaaaacttg gccctcactg acagatgagg ggcggacgtt gacacttgag
    16081 gggccgactc acccggcgcg gcgttgacag atgaggggca ggctcgattt cggccggcga
    16141 cgtggagctg gccagcctcg caaatcggcg aaaacgcctg attttacgcg agtttcccac
    16201 agatgatgtg gacaagcctg gggataagtg ccctgcggta ttgacacttg aggggcgcga
    16261 ctactgacag atgaggggcg cgatccttga cacttgaggg gcagagtgct gacagatgag
    16321 gggcgcacct attgacattt gaggggctgt ccacaggcag aaaatccagc atttgcaagg
    16381 gtttccgccc gtttttcggc caccgctaac ctgtctttta acctgctttt aaaccaatat
    16441 ttataaacct tgtttttaac cagggctgcg ccctgtgcgc gtgaccgcgc acgccgaagg
    16501 ggggtgcccc cccttctcga accctcccgg tcgagtcatg gtctagcatg gatctcgggg
    16561 acgtcaccca taatacccat aatagctgtt tgccagtact ttgatcccga ggggaaccct
    16621 gtggttggca tgcacataca aatggacgaa cggataaacc ttttcacgcc cttttaaata
    16681 tccgttattc taataaacgc tcttttctct tag
//
