LOCUS       sequence-477275-       18366 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plant expression of Cas9 and gRNAs against  A. thaliana CK2alpha3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 18366)
  TITLE     Hothorn CK2 plasmids
REFERENCE   2  (bases 1 to 18366)
  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"
COMMENT     Sequence Label: pLIII CKBs CRISPR with guides at 1-2 and 2-3
FEATURES             Location/Qualifiers
     source          1..18366
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      316..510
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(576..1649)
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGASSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     CDS             complement(2078..2707)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    2971..2995
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_RNA        3293..3368
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        3620..3695
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        3947..4022
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        4274..4349
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        4601..4676
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        4928..5003
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        5263..5338
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        5590..5665
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        5917..5992
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        6244..6319
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        6571..6646
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        6898..6973
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             8739..8804
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             8811..8831
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8856..12956
                     /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             8856..12956
                     /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             12957..13004
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      13018..13266
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     misc_feature    16458..16482
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(16605..17193)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(16685..16704)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(17331..17350)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(17332..17350)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(17355..17371)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(17355..17371)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(17473..17492)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             17484..18293
                     /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
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(17551..17570)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc tgccattttt
       61 ggggtgaggc cgttcgcggc cgaggggcgc agcccctggg gggatgggag ctcggtgcct
      121 tgatgtgggc gccggcggtc gagtggcgac ggcgcggctt gtccgcgccc tggtagattg
      181 cctggccgta ggccagccat ttttgagcgg ccagcggccg cgataggccg acgcgaagcg
      241 gcggggcgta gggagcgcag cgaccgaagg gtaggcgctt tttgcagctc ttcggctgtg
      301 cgctggccag acagttatgc acaggccagg cgggttttaa gagttttaat aagttttaaa
      361 gagttttagg cggaaaaatc gccttttttc tcttttatat cagtcactta catgtgtgac
      421 cggttcccaa tgtacggctt tgggttccca atgtacgggt tccggttccc aatgtacggc
      481 tttgggttcc caatgtacgt gctatccaca ggaaagaagc tttttcgacc tttttcccct
      541 gctagggcaa tttgccctag catctgctcc gtacattagg aaccggcgga tgcttcgccc
      601 tcgatcaggt tgcggtagcg catgactagg atcgggccag cctgccccgc ctcctccttc
      661 aaatcgtact ccggcaggtc atttgacccg atcagcttgc gcacggtgaa acagaacttc
      721 ttgaactctc cggcgctgcc actgcgttcg tagatcgtct tgaacaacca tctggcttct
      781 gccttgcctg cggcgcggcg tgccaggcgg tagagaaaac ggccgatgcc gggatcgatc
      841 aaaaagtaat cggggtgaac cgtcagcacg tccgggttct tgccttctgt gatctcgcgg
      901 tacatccaat cagctagctc gatctcgatg tactccggcc gcccggtttc gctctttacg
      961 atcttgtagc ggctaatcaa ggcttcaccc tcggataccg tcaccaggcg gccgttcttg
     1021 gccttcttcg tacgctgcat ggcaacgtgc gtggtgttta accgaatgca ggtttctacc
     1081 aggtcgtctt tctgctttcc gccatcggct cgccggcaga acttgagtac gtccgcaacg
     1141 tgtggacgga acacgcggcc gggcttgtct cccttccctt cccggtatcg gttcatggat
     1201 tcggttagat gggaaaccgc catcagtacc aggtcgtaat cccacacact ggccatgccg
     1261 gccggccctg cggaaacctc tacgtgcccg tctggaagct cgtagcggat cacctcgcca
     1321 gctcgtcggt cacgcttcga cagacggaaa acggccacgt ccatgatgct gcgactatcg
     1381 cgggtgccca cgtcatagag catcggaacg aaaaaatctg gttgctcgtc gcccttgggc
     1441 ggcttcctaa tcgacgacgc accggctgcc ggcggttgcc gggattcttt gcggattcga
     1501 tcagcggccg cttgccacga ttcaccgggg cgtgcttctg cctcgatgcg ttgccgctgg
     1561 gcggcctgcg cggccttcaa cttctccacc aggtcatcac ccagcgccgc gccgatttgt
     1621 accgggccgg atggtttgcg accgctcacg ccgattcctc gggcttgggg gttccagtgc
     1681 cattgcaggg ccggcagaca acccagccgc ttacgcctgg ccaaccgccc gttcctccac
     1741 acatggggca ttccacggcg tcggtgcctg gttgttcttg attttccatg ccgcctcctt
     1801 tagccgctaa aattcatcta ctcatttatt catttgctca tttactctgg tagctgcgcg
     1861 atgtattcag atagcagctc ggtaatggtc ttgccttggc gtaccgcgta catcttcagc
     1921 ttggtgtgat cctccgccgg caactgaaag ttgacccgct tcatggctgg cgtgtctgcc
     1981 aggctggcca acgttgcagc cttgctgctg cgtgcgctcg gacggccggc acttagcgtg
     2041 tttgtgcttt tgctcatttt ctctttacct cattaactca aatgagtttt gatttaattt
     2101 cagcggccag cgcctggacc tcgcgggcag cgtcgccctc gggttctgat tcaagaacgg
     2161 ttgtgccggc ggcggcagtg cctgggtagc tcacgcgctg cgtgatacgg gactcaagaa
     2221 tgggcagctc gtacccggcc agcgcctcgg caacctcacc gccgatgcgc gtgcctttga
     2281 tcgcccgcga cacgacaaag gccgcttgta gccttccatc cgtgacctca atgcgctgct
     2341 taaccagctc caccaggtcg gcggtggccc atatgtcgta agggcttggc tgcaccggaa
     2401 tcagcacgaa gtcggctgcc ttgatcgcgg acacagccaa gtccgccgcc tggggcgctc
     2461 cgtcgatcac tacgaagtcg cgccggccga tggccttcac gtcgcggtca atcgtcgggc
     2521 ggtcgatgcc gacaacggtt agcggttgat cttcccgcac ggccgcccaa tcgcgggcac
     2581 tgccctgggg atcggaatcg actaacagaa catcggcccc ggcgagttgc agggcgcggg
     2641 ctagatgggt tgcgatggtc gtcttgcctg acccgccttt ctggttaagt acagcgataa
     2701 ccttcatgcg ttccccttgc gtatttgttt atttactcat cgcatcatat acgcagcgac
     2761 cgcatgacgc aagctgtttt actcaaatac acatcacctt tttagacggc ggcgctcggt
     2821 ttcttcagcg gccaagctgg ccggccaggc cgccagcttg gcatcagaca aaccggccag
     2881 gatttcatgc agccgcacgg ctgatgggct gcctgtatcg agtggtgatt ttgtgccgag
     2941 ctgccggtcg gggagctgtt ggctggctgg tggcaggata tattgtggtg taaacaaatt
     3001 gacgcttaga caacttaata acacattgcg gacgttttta atgtactggg gtggattacg
     3061 gcgggggcat cttcattctt aagatatgaa gataatcttc aaaaggcccc tgggaatctg
     3121 aaagaagaga agcaggccca tttatatggg aaagaacaat agtatttctt atataggccc
     3181 atttaagttg aaaacaatct tcaaaagtcc cacatcgctt agataagaaa acgaagctga
     3241 gtttatatac agctagagtc gaagtagtga ttggtaagtc tgattgacca acgttttaga
     3301 gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga
     3361 gtcggtgctt ttttttactg tcgagtatct ggggcatctt cattcttaag atatgaagat
     3421 aatcttcaaa aggcccctgg gaatctgaaa gaagagaagc aggcccattt atatgggaaa
     3481 gaacaatagt atttcttata taggcccatt taagttgaaa acaatcttca aaagtcccac
     3541 atcgcttaga taagaaaacg aagctgagtt tatatacagc tagagtcgaa gtagtgattg
     3601 tgagagattg agaaatcgag ttttagagct agaaatagca agttaaaata aggctagtcc
     3661 gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt tttactgtcg agtacaccgg
     3721 gcatcttcat tcttaagata tgaagataat cttcaaaagg cccctgggaa tctgaaagaa
     3781 gagaagcagg cccatttata tgggaaagaa caatagtatt tcttatatag gcccatttaa
     3841 gttgaaaaca atcttcaaaa gtcccacatc gcttagataa gaaaacgaag ctgagtttat
     3901 atacagctag agtcgaagta gtgattgaga cgtgtgtgaa cagaaagttt tagagctaga
     3961 aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt
     4021 gctttttttt actgtcgagt aaagggggca tcttcattct taagatatga agataatctt
     4081 caaaaggccc ctgggaatct gaaagaagag aagcaggccc atttatatgg gaaagaacaa
     4141 tagtatttct tatataggcc catttaagtt gaaaacaatc ttcaaaagtc ccacatcgct
     4201 tagataagaa aacgaagctg agtttatata cagctagagt cgaagtagtg attgcagaac
     4261 caagatatcc acggttttag agctagaaat agcaagttaa aataaggcta gtccgttatc
     4321 aacttgaaaa agtggcaccg agtcggtgct tttttttact gtcgagtaaa tcgggcatct
     4381 tcattcttaa gatatgaaga taatcttcaa aaggcccctg ggaatctgaa agaagagaag
     4441 caggcccatt tatatgggaa agaacaatag tatttcttat ataggcccat ttaagttgaa
     4501 aacaatcttc aaaagtccca catcgcttag ataagaaaac gaagctgagt ttatatacag
     4561 ctagagtcga agtagtgatt ggagagagag atgagctcta gttttagagc tagaaatagc
     4621 aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt
     4681 ttttactgtc gagtatgagg ggcatcttca ttcttaagat atgaagataa tcttcaaaag
     4741 gcccctggga atctgaaaga agagaagcag gcccatttat atgggaaaga acaatagtat
     4801 ttcttatata ggcccattta agttgaaaac aatcttcaaa agtcccacat cgcttagata
     4861 agaaaacgaa gctgagttta tatacagcta gagtcgaagt agtgattgga aaatcgaatc
     4921 taaccgtgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg
     4981 aaaaagtggc accgagtcgg tgcttttttt tactgtcgag tatgtcgggc gcgggggcat
     5041 cttcattctt aagatatgaa gataatcttc aaaaggcccc tgggaatctg aaagaagaga
     5101 agcaggccca tttatatggg aaagaacaat agtatttctt atataggccc atttaagttg
     5161 aaaacaatct tcaaaagtcc cacatcgctt agataagaaa acgaagctga gtttatatac
     5221 agctagagtc gaagtagtga ttgctctcat cctctaaccc aagttttaga gctagaaata
     5281 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
     5341 ttttttactg tcgagtatct ggggcatctt cattcttaag atatgaagat aatcttcaaa
     5401 aggcccctgg gaatctgaaa gaagagaagc aggcccattt atatgggaaa gaacaatagt
     5461 atttcttata taggcccatt taagttgaaa acaatcttca aaagtcccac atcgcttaga
     5521 taagaaaacg aagctgagtt tatatacagc tagagtcgaa gtagtgattg atatactgcc
     5581 ctaaatgcgg ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact
     5641 tgaaaaagtg gcaccgagtc ggtgcttttt tttactgtcg agtacaccgg gcatcttcat
     5701 tcttaagata tgaagataat cttcaaaagg cccctgggaa tctgaaagaa gagaagcagg
     5761 cccatttata tgggaaagaa caatagtatt tcttatatag gcccatttaa gttgaaaaca
     5821 atcttcaaaa gtcccacatc gcttagataa gaaaacgaag ctgagtttat atacagctag
     5881 agtcgaagta gtgattgcaa cagcctggct cattgggttt tagagctaga aatagcaagt
     5941 taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gctttttttt
     6001 actgtcgagt aaagggggca tcttcattct taagatatga agataatctt caaaaggccc
     6061 ctgggaatct gaaagaagag aagcaggccc atttatatgg gaaagaacaa tagtatttct
     6121 tatataggcc catttaagtt gaaaacaatc ttcaaaagtc ccacatcgct tagataagaa
     6181 aacgaagctg agtttatata cagctagagt cgaagtagtg attgatatac tgcccaaaat
     6241 gcggttttag agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa
     6301 agtggcaccg agtcggtgct tttttttact gtcgagtaaa tcgggcatct tcattcttaa
     6361 gatatgaaga taatcttcaa aaggcccctg ggaatctgaa agaagagaag caggcccatt
     6421 tatatgggaa agaacaatag tatttcttat ataggcccat ttaagttgaa aacaatcttc
     6481 aaaagtccca catcgcttag ataagaaaac gaagctgagt ttatatacag ctagagtcga
     6541 agtagtgatt gataaagctt ttttgaacag gttttagagc tagaaatagc aagttaaaat
     6601 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttttactgtc
     6661 gagtatgagg ggcatcttca ttcttaagat atgaagataa tcttcaaaag gcccctggga
     6721 atctgaaaga agagaagcag gcccatttat atgggaaaga acaatagtat ttcttatata
     6781 ggcccattta agttgaaaac aatcttcaaa agtcccacat cgcttagata agaaaacgaa
     6841 gctgagttta tatacagcta gagtcgaagt agtgattgga tgcagttgaa tctgacagtt
     6901 ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc
     6961 accgagtcgg tgcttttttt tactgtcgag tatgtcattg tagataatta ggctaaacta
     7021 tccttaaggt gtctagataa ttaggctaaa ctatccttaa ggtgaggcgg atccctcaac
     7081 ttttgattcg ctatttgcag tgcacctgtg gcgttcatca catcttttgt gacactgttt
     7141 gcactggtca ttgctattac aaaggacctt cctgatgttg aaggagatcg aaagtaagta
     7201 actgcacgca taaccatttt ctttccgctc tttggctcaa tccatttgac agtcaaagac
     7261 aatgtttaac cagctccgtt tgatatattg tctttatgtg tttgttcaag catgtttagt
     7321 taatcatgcc tttgattgat cttgaatagg ttccaaatat caaccctggc aacaaaactt
     7381 ggagtgagaa acattgcatt cctcggttct ggacttctgc tagtaaatta tgtttcagcc
     7441 atatcactag ctttctacat gcctcaggtg aattcatcta tttccgtctt aactatttcg
     7501 gttaattaaa gcacgaacac cattactgca tgtagaagct tgataaacta tcgccaccaa
     7561 tttatttttg ttgcgatatt gttactttcc tcagtatgca gctttgaaaa gaccaaccct
     7621 cttatccttt aacaatgaac aggtttttag aggtagcttg atgattcctg cacatgtgat
     7681 cttggcttca ggcttaattt tccaggtaaa gcattatgag atactcttat atctcttaca
     7741 tacttttgag ataatgcaca agaacttcat aactatatgc tttagtttct gcatttgaca
     7801 ctgccaaatt cattaatctc taatatcttt gttgttgatc tttggtagac atgggtacta
     7861 gaaaaagcaa actacaccaa ggtaaaatac ttttgtacaa acataaactc gttatcacgg
     7921 aacatcaatg gagtgtatat ctaacggagt gtagaaacat ttgattattg caggaagcta
     7981 tctcaggata ttatcggttt atatggaatc tcttctacgc agagtatctg ttattcccct
     8041 tcctctagct ttcaatttca tggtgaggat atgcagtttt ctttgtatat cattcttctt
     8101 cttctttgta gcttggagtc aaaatcggtt ccttcatgta catacatcaa ggatatgtcc
     8161 ttctgaattt ttatatcttg caataaaaat gcttgtacca attgaaacac cagctttttg
     8221 agttctatga tcactgactt ggttctaacc aaaaaaaaaa aatgtttaat ttacatatct
     8281 aaaagtaggt ttagggaaac ctaaacagta aaatatttgt atattattcg aatttcactc
     8341 atcataaaaa cttaaattgc accataaaat tttgttttac tattaatgat gtaatttgtg
     8401 taacttaaga taaaaataat attccgtaag ttaaccggct aaaaccacgt ataaaccagg
     8461 gaacctgtta aaccggttct ttactggata aagaaatgaa agcccatgta gacagctcca
     8521 ttagagccca aaccctaaat ttctcatcta tataaaagga gtgacattag ggtttttgtt
     8581 cgtcctctta aagcttctcg ttttctctgc cgtctctctc attcgcgcga cgcaaacgat
     8641 cttcaggtga tcttctttct ccaaatcctc tctcataact ctgatttcgt acttgtgtat
     8701 ttgagctcac gctctgtttc tctcaccaca gccccatgga ctataaggac cacgacggag
     8761 actacaagga tcatgatatt gattacaaag acgatgacga taagatggcc ccaaagaaga
     8821 agcggaaggt cggtatccac ggagtcccag cagccgacaa gaagtacagc atcggcctgg
     8881 acatcggcac caactctgtg ggctgggccg tgatcaccga cgagtacaag gtgcccagca
     8941 agaaattcaa ggtgctgggc aacaccgacc ggcacagcat caagaagaac ctgatcggag
     9001 ccctgctgtt cgacagcggc gaaacagccg aggccacccg gctgaagaga accgccagaa
     9061 gaagatacac cagacggaag aaccggatct gctatctgca agagatcttc agcaacgaga
     9121 tggccaaggt ggacgacagc ttcttccaca gactggaaga gtccttcctg gtggaagagg
     9181 ataagaagca cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg
     9241 agaagtaccc caccatctac cacctgagaa agaaactggt ggacagcacc gacaaggccg
     9301 acctgcggct gatctatctg gccctggccc acatgatcaa gttccggggc cacttcctga
     9361 tcgagggcga cctgaacccc gacaacagcg acgtggacaa gctgttcatc cagctggtgc
     9421 agacctacaa ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg
     9481 ccatcctgtc tgccagactg agcaagagca gacggctgga aaatctgatc gcccagctgc
     9541 ccggcgagaa gaagaatggc ctgttcggaa acctgattgc cctgagcctg ggcctgaccc
     9601 ccaacttcaa gagcaacttc gacctggccg aggatgccaa actgcagctg agcaaggaca
     9661 cctacgacga cgacctggac aacctgctgg cccagatcgg cgaccagtac gccgacctgt
     9721 ttctggccgc caagaacctg tccgacgcca tcctgctgag cgacatcctg agagtgaaca
     9781 ccgagatcac caaggccccc ctgagcgcct ctatgatcaa gagatacgac gagcaccacc
     9841 aggacctgac cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag tacaaagaga
     9901 ttttcttcga ccagagcaag aacggctacg ccggctacat tgacggcgga gccagccagg
     9961 aagagttcta caagttcatc aagcccatcc tggaaaagat ggacggcacc gaggaactgc
    10021 tcgtgaagct gaacagagag gacctgctgc ggaagcagcg gaccttcgac aacggcagca
    10081 tcccccacca gatccacctg ggagagctgc acgccattct gcggcggcag gaagattttt
    10141 acccattcct gaaggacaac cgggaaaaga tcgagaagat cctgaccttc cgcatcccct
    10201 actacgtggg ccctctggcc aggggaaaca gcagattcgc ctggatgacc agaaagagcg
    10261 aggaaaccat caccccctgg aacttcgagg aagtggtgga caagggcgct tccgcccaga
    10321 gcttcatcga gcggatgacc aacttcgata agaacctgcc caacgagaag gtgctgccca
    10381 agcacagcct gctgtacgag tacttcaccg tgtataacga gctgaccaaa gtgaaatacg
    10441 tgaccgaggg aatgagaaag cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg
    10501 acctgctgtt caagaccaac cggaaagtga ccgtgaagca gctgaaagag gactacttca
    10561 agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct
    10621 ccctgggcac ataccacgat ctgctgaaaa ttatcaagga caaggacttc ctggacaatg
    10681 aggaaaacga ggacattctg gaagatatcg tgctgaccct gacactgttt gaggacagag
    10741 agatgatcga ggaacggctg aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc
    10801 agctgaagcg gcggagatac accggctggg gcaggctgag ccggaagctg atcaacggca
    10861 tccgggacaa gcagtccggc aagacaatcc tggatttcct gaagtccgac ggcttcgcca
    10921 acagaaactt catgcagctg atccacgacg acagcctgac ctttaaagag gacatccaga
    10981 aagcccaggt gtccggccag ggcgatagcc tgcacgagca cattgccaat ctggccggca
    11041 gccccgccat taagaagggc atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag
    11101 tgatgggccg gcacaagccc gagaacatcg tgatcgaaat ggccagagag aaccagacca
    11161 cccagaaggg acagaagaac agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag
    11221 agctgggcag ccagatcctg aaagaacacc ccgtggaaaa cacccagctg cagaacgaga
    11281 agctgtacct gtactacctg cagaatgggc gggatatgta cgtggaccag gaactggaca
    11341 tcaaccggct gtccgactac gatgtggacc atatcgtgcc tcagagcttt ctgaaggacg
    11401 actccatcga caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg
    11461 tgccctccga agaggtcgtg aagaagatga agaactactg gcggcagctg ctgaacgcca
    11521 agctgattac ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg
    11581 aactggataa ggccggcttc atcaagagac agctggtgga aacccggcag atcacaaagc
    11641 acgtggcaca gatcctggac tcccggatga acactaagta cgacgagaat gacaagctga
    11701 tccgggaagt gaaagtgatc accctgaagt ccaagctggt gtccgatttc cggaaggatt
    11761 tccagtttta caaagtgcgc gagatcaaca actaccacca cgcccacgac gcctacctga
    11821 acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt
    11881 acggcgacta caaggtgtac gacgtgcgga agatgatcgc caagagcgag caggaaatcg
    11941 gcaaggctac cgccaagtac ttcttctaca gcaacatcat gaactttttc aagaccgaga
    12001 ttaccctggc caacggcgag atccggaagc ggcctctgat cgagacaaac ggcgaaaccg
    12061 gggagatcgt gtgggataag ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc
    12121 cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc aaagagtcta
    12181 tcctgcccaa gaggaacagc gataagctga tcgccagaaa gaaggactgg gaccctaaga
    12241 agtacggcgg cttcgacagc cccaccgtgg cctattctgt gctggtggtg gccaaagtgg
    12301 aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct gctggggatc accatcatgg
    12361 aaagaagcag cttcgagaag aatcccatcg actttctgga agccaagggc tacaaagaag
    12421 tgaaaaagga cctgatcatc aagctgccta agtactccct gttcgagctg gaaaacggcc
    12481 ggaagagaat gctggcctct gccggcgaac tgcagaaggg aaacgaactg gccctgccct
    12541 ccaaatatgt gaacttcctg tacctggcca gccactatga gaagctgaag ggctcccccg
    12601 aggataatga gcagaaacag ctgtttgtgg aacagcacaa gcactacctg gacgagatca
    12661 tcgagcagat cagcgagttc tccaagagag tgatcctggc cgacgctaat ctggacaaag
    12721 tgctgtccgc ctacaacaag caccgggata agcccatcag agagcaggcc gagaatatca
    12781 tccacctgtt taccctgacc aatctgggag cccctgccgc cttcaagtac tttgacacca
    12841 ccatcgaccg gaagaggtac accagcacca aagaggtgct ggacgccacc ctgatccacc
    12901 agagcatcac cggcctgtac gagacacgga tcgacctgtc tcagctggga ggcgacaaaa
    12961 ggccggcggc cacgaaaaag gccggccagg caaaaaagaa aaagtaaaag ggtgggatat
    13021 gaagatgaag atgaaatatt tggtgtgtca aataaaaagc ttgtgtgctt aagtttgtgt
    13081 ttttttcttg gcttgttgtg ttatgaattt gtggcttttt ctaatattaa atgaatgtaa
    13141 gatctcatta taatgaataa acaaatgttt ctataatcca ttgtgaatgt tttgttggat
    13201 ctcttctgca gcatataact actgtatgtg ctatggtatg gactatggaa tatgattaaa
    13261 gataagtctg gaagcacctg acctaagcta gcctagaagg gctaagacct aagctagcct
    13321 agaatctgac ctaagctagc ctagtgaggt atgtaggtat agtaacatga acaagaacag
    13381 actcaagtac aagatcgcat acgaaaatgg aaatggcaat gtcacttcca cataatcaaa
    13441 cacgaatcct catatcaaca aggcctgaga ttctaactag ctcataacaa cttagccaat
    13501 agttacttga gactaccaaa tgtatgtaga actaaagact aagggacaga gagttcgtct
    13561 aaacaggtga atctagtcgt tgttatctaa taaacaattc agccccaaat gcagaacaca
    13621 catagagctc tctattgatt caaattacga tctgatactg ataacgtcta gatttttagg
    13681 gttaaagcaa tcaatcacct gacgattcaa ggtggttgga tcatgacgat tccagaaaac
    13741 atcaagcaag ctctcaaagc tacactcttt gggatcatac tgaactctaa caacctcgtt
    13801 atgtcccgta gtgccagtac agacatcctc gtaactcgga ttgtgcacga tgccatggct
    13861 atacccaacc tcggtcttgg tcacaccagg aactctctgg taagctagct ccactcccca
    13921 gaaacaaccg gcgccaaatt gcgcgaattg ctgacctgat gacggaacat catcgtcggg
    13981 tccttgggcg attgcggcgg aagatgggtc agcttgggct tgaggacgag agccgaatcc
    14041 gagtctgttg aaaaggttgt tcattgggga tttgtatacg gagattggtc gtcgagaggt
    14101 ttgagggaaa ggacaaatgg gtttggctct ggagaaagag agtgcggctt tagagagaga
    14161 attgagaggt ttagagagag atgcggcggc gatgagcgtg ccgaggagag acgacgagga
    14221 cctgcattat caaagcagtg acgtggtgaa atttggaact tttaagaggc agatagattt
    14281 attatttgta tccattttct tcattgttct agaatgtcgc ggaacaaatt ttaaaactaa
    14341 atcctaaatt tttctaattt tgttgccaat agtggatatg tgggccgtat agaaggaatc
    14401 tattgaaggc tcaaacccat actgacgagc ccaaaggttc gttttgcgtt ttatgtttcg
    14461 gttcgatgcc aacgccacat tctgagctag gcaaaaaaca aacgtgtctt tgaatagact
    14521 cctctcgtta acacatgcag cggctgcatg gtgacgccat taacacgtgg cctacaattg
    14581 catgatgtct ccattgacac gtgacttctc gtctcctttc ttaatatatc taacaaacac
    14641 tcctacctct tccaaaatat atacacatct ttttgatcaa tctctcattc aaaatctcat
    14701 tctctctagt aaacaagaac aaaaaaatgg cggatacagc tagaggaacc catcacgata
    14761 tcatcggcag ggaccagtac ccgatgatgg gccgtgaccg tgaccagtac cagatgtccg
    14821 gacgaggatc tgactactcc aagtctaggc agattgctaa agctgcaact gctgtcacag
    14881 ctggtggttc cctccttgtt ctctccagcc ttacccttgt tggaactgtc atagctttga
    14941 ctgttgcaac acctctgctc gttatcttca gcccaatcct tgtcccggct ctcatcacag
    15001 ttgcactcct catcaccggt tttctttcct ctggagggtt tggcattgcc gctataaccg
    15061 ttttctcttg gatttacaag taagcacaca tttatcatct tacttcataa ttttgtgcaa
    15121 tatgtgcatg catgtgttga gccagtagct ttggatcaat ttttttggtc gaataacaaa
    15181 tgtaacaata agaaattgca aattctaggg aacatttggt taactaaata cgaaatttga
    15241 cctagctagc ttgaatgtgt ctgtgtatat catctatata ggtaaaatgc ttggtatgat
    15301 acctattgat tgtgaatagg tacgcaacgg gagagcaccc acagggatca gacaagttgg
    15361 acagtgcaag gatgaagttg ggaagcaaag ctcaggatct gaaagacaga gctcagtact
    15421 acggacagca acatactggt ggggaacatg accgtgaccg tactcgtggt ggccagcaca
    15481 ctactatgag cgagctgatt aaggagaaca tgcacatgaa gctgtacatg gagggcaccg
    15541 tgaacaacca ccacttcaag tgcacatccg agggcgaagg caagccctac gagggcaccc
    15601 agaccatgag aatcaaggtg gtcgagggcg gccctctccc cttcgccttc gacatcctgg
    15661 ctaccagctt catgtacggc agcagaacct tcatcaacca cacccagggc atccccgact
    15721 tctttaagca gtccttccct gagggcttca catgggagag agtcaccaca tacgaggacg
    15781 gaggtgtgct gaccgctact caggacacca gcctccagga cggctgcctc atctacaacg
    15841 tcaagatcag aggggtgaac ttcccatcca acggccctgt gatgcagaag aaaacactcg
    15901 gctgggaagc caacaccgag atgctgtatc cagctgacgg aggactggaa ggcagaagcg
    15961 acatggccct gaagctcgtg ggcgggggcc acctgatctg caacttcaag accacataca
    16021 gatccaagaa acccgctaag aacctcaaga tgcccggcgt ctactatgtg gaccacagac
    16081 tggaaagaat caaggaggcc gacaaagaaa cctacgtcga gcagcacgag gtggctgtgg
    16141 ccagatactg cgacctccct agcaaactgg ggcacaagtg attagagttt ttcagatccg
    16201 cgtttgtgtt ttctgggttt ctcacttaag cgtctgcgtt ttacttttgt attgggtttg
    16261 gcgtttagta gtttgcggta gcgttcttgt tatgtgtaat tacgcttttt cttcttgctt
    16321 cagcagtttc ggttgaaata taaatcgaat caagtttcac tttatcagcg ttgttttaaa
    16381 ttttggcatt aaattggtga aaattgcttc aattttgtat ctaaatagaa gtgtctctgg
    16441 tttaaactat cagtgtttga caggatatat tggcgggtaa acctaagaga aaagagcgtt
    16501 tattagaata atcggatatt taaaagggcg tgaaaaggtt tatccgttcg tccatttgta
    16561 tgtgccaacc acaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
    16621 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
    16681 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
    16741 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
    16801 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
    16861 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
    16921 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
    16981 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
    17041 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
    17101 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
    17161 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
    17221 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
    17281 aggatcttca cctagatcat aaaatcatta tttgccatcc agctgatatc ccctatagtg
    17341 agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct
    17401 gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca
    17461 taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat
    17521 taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc
    17581 aatcaggtgc gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga
    17641 aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc
    17701 tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat
    17761 ggttactcac cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg
    17821 attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc
    17881 ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tcttgctcag gcgcaatcac
    17941 gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg
    18001 ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca
    18061 ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta
    18121 ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact
    18181 gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata
    18241 atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat
    18301 tggttaattg gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcgcaagct
    18361 catgac
//
