LOCUS       sequence-477173-       16640 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plant expression of Cas9 and gRNAs against A. thaliana CK2alpha4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16640)
  TITLE     Hothorn CK2 plasmids
REFERENCE   2  (bases 1 to 16640)
  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: pLII-CRISPR for CKA4
FEATURES             Location/Qualifiers
     source          1..16640
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      53..641
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     542..561
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     795..812
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    827..967
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(893..912)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1053..1075
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      1311..1505
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(1571..2644)
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     CDS             complement(3073..3702)
                     /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    5003..5027
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             6734..6799
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             6806..6826
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6851..10951
                     /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             6851..10951
                     /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             10952..10999
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      11013..11261
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     misc_RNA        11502..11577
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        11855..11930
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    15077..15101
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             complement(15479..16270)
                     /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"
ORIGIN
        1 tgccgttctt ccgatgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
       61 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
      121 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
      181 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
      241 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
      301 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
      361 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
      421 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
      481 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
      541 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
      601 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
      661 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
      721 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
      781 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
      841 ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct
      901 gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat
      961 ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     1021 ggcatccgct tacagacaag ctgtgactgt ctccgggagc tgcatgtgtc agaggttttc
     1081 accgtcatca ccgaaacgcg cgaggcaggg tgccttgatg tgggcgccgg cggtcgagtg
     1141 gcgacggcgc ggcttgtccg cgccctggta gattgcctgg ccgtaggcca gccatttttg
     1201 agcggccagc ggccgcgata ggccgacgcg aagcggcggg gcgtagggag cgcagcgacc
     1261 gaagggtagg cgctttttgc agctcttcgg ctgtgcgctg gccagacagt tatgcacagg
     1321 ccaggcgggt tttaagagtt ttaataagtt ttaaagagtt ttaggcggaa aaatcgcctt
     1381 ttttctcttt tatatcagtc acttacatgt gtgaccggtt cccaatgtac ggctttgggt
     1441 tcccaatgta cgggttccgg ttcccaatgt acggctttgg gttcccaatg tacgtgctat
     1501 ccacaggaaa gagtcctttt cgaccttttt cccctgctag ggcaatttgc cctagcatct
     1561 gctccgtaca ttaggaaccg gcggatgctt cgccctcgat caggttgcgg tagcgcatga
     1621 ctaggatcgg gccagcctgc cccgcctcct ccttcaaatc gtactccggc aggtcatttg
     1681 acccgatcag cttgcgcacg gtgaaacaga acttcttgaa ctctccggcg ctgccactgc
     1741 gttcgtagat cgtcttgaac aaccatctgg cttctgcctt gcctgcggcg cggcgtgcca
     1801 ggcggtagag aaaacggccg atgccgggat cgatcaaaaa gtaatcgggg tgaaccgtca
     1861 gcacgtccgg gttcttgcct tctgtgatct cgcggtacat ccaatcagct agctcgatct
     1921 cgatgtactc cggccgcccg gtttcgctct ttacgatctt gtagcggcta atcaaggctt
     1981 caccctcgga taccgtcacc aggcggccgt tcttggcctt cttcgtacgc tgcatggcaa
     2041 cgtgcgtggt gtttaaccga atgcaggttt ctaccaggtc gtctttctgc tttccgccat
     2101 cggctcgccg gcagaacttg agtacgtccg caacgtgtgg acggaacacg cggccgggct
     2161 tgtctccctt cccttcccgg tatcggttca tggattcggt tagatgggaa accgccatca
     2221 gtaccaggtc gtaatcccac acactggcca tgccggccgg ccctgcggaa acctctacgt
     2281 gcccgtctgg aagctcgtag cggatcacct cgccagctcg tcggtcacgc ttcgacagac
     2341 ggaaaacggc cacgtccatg atgctgcgac tatcgcgggt gcccacgtca tagagcatcg
     2401 gaacgaaaaa atctggttgc tcgtcgccct tgggcggctt cctaatcgac ggcgcaccgg
     2461 ctgccggcgg ttgccgggat tctttgcgga ttcgatcagc ggccgcttgc cacgattcac
     2521 cggggcgtgc ttctgcctcg atgcgttgcc gctgggcggc ctgcgcggcc ttcaacttct
     2581 ccaccaggtc atcacccagc gccgcgccga tttgtaccgg gccggatggt ttgcgaccgc
     2641 tcacgccgat tcctcgggct tgggggttcc agtgccattg cagggccggc aggcaaccca
     2701 gccgcttacg cctggccaac cgcccgttcc tccacacatg gggcattcca cggcgtcggt
     2761 gcctggttgt tcttgatttt ccatgccgcc tcctttagcc gctaaaattc atctactcat
     2821 ttattcattt gctcatttac tctggtagct gcgcgatgta ttcagatagc agctcggtaa
     2881 tggtcttgcc ttggcgtacc gcgtacatct tcagcttggt gtgatcctcc gccggcaact
     2941 gaaagttgac ccgcttcatg gctggcgtgt ctgccaggct ggccaacgtt gcagccttgc
     3001 tgctgcgtgc gctcggacgg ccggcactta gcgtgtttgt gcttttgctc attttctctt
     3061 tacctcatta actcaaatga gttttgattt aatttcagcg gccagcgcct ggacctcgcg
     3121 ggcagcgtcg ccctcgggtt ctgattcaag aacggttgtg ccggcggcgg cagtgcctgg
     3181 gtagctcacg cgctgcgtga tacgggactc aagaatgggc agctcgtacc cggccagcgc
     3241 ctcggcaacc tcaccgccga tgcgcgtgcc tttgatcgcc cgcgacacga caaaggccgc
     3301 ttgtagcctt ccatccgtga cctcaatgcg ctgcttaacc agctccacca ggtcggcggt
     3361 ggcccatatg tcgtaagggc ttggctgcac cggaatcagc acgaagtcgg ctgccttgat
     3421 cgcggacaca gccaagtccg ccgcctgggg cgctccgtcg atcactacga agtcgcgccg
     3481 gccgatggcc ttcacgtcgc ggtcaatcgt cgggcggtcg atgccgacaa cggttagcgg
     3541 ttgatcttcc cgcacggccg cccaatcgcg ggcactgccc tggggatcgg aatcgactaa
     3601 cagaacatcg gccccggcga gttgcagggc gcgggctaga tgggttgcga tggtcgtctt
     3661 gcctgacccg cctttctggt taagtacagc gataaccttc atgcgttccc cttgcgtatt
     3721 tgtttattta ctcatcgcat catatacgca gcgaccgcat gacgcaagct gttttactca
     3781 aatacacatc acctttttag acggcggcgc tcggtttctt cagcggccaa gctggccggc
     3841 caggccgcca gcttggcatc agacaaaccg gccaggattt catgcagccg cacggttgag
     3901 atgtgcgcgg gcggctcgaa cacgtacccg gccgcgatca tctccgcctc gatctcttcg
     3961 gtaatgaaaa acggttcgtc ctggccgtcc tggtgcggtt tcatgcttgt tcctcttggc
     4021 gttcattctc ggcggccgcc agggcgtcgg cctcggtcaa tgcgtcctca cggaaggcac
     4081 cgcgccgcct ggcctcggtg ggcgtcactt cctcgctgcg ctcaagtgcg cggtacaggg
     4141 tcgagcgatg cacgccaagc agtgcagccg cctctttcac ggtgcggcct tcctggtcga
     4201 tcagctcgcg ggcgtgcgcg atctgtgccg gggtgagggt agggcggggg ccaaacttca
     4261 cgcctcgggc cttggcggcc tcgcgcccgc tccgggtgcg gtcgatgatt agggaacgct
     4321 cgaactcggc aatgccggcg aacacggtca acaccatgcg gccggccggc gtggtggtgt
     4381 cggcccacgg ctctgccagg ctacgcaggc ccgcgccggc ctcctggatg cgctcggcaa
     4441 tgtccagtag gtcgcgggtg ctgcgggcca ggcggtctag cctggtcact gtcacaacgt
     4501 cgccagggcg taggtggtca agcatcctgg ccagctccgg gcggtcgcgc ctggtgccgg
     4561 tgatcttctc ggaaaacagc ttggtgcagc cggccgcgtg cagttcggcc cgttggttgg
     4621 tcaagtcctg gtcgtcggtg ctgacgcggg catagcccag caggccagcg gcggcgctct
     4681 tgttcatggc gtaatgtctc cggttctagt cgcaagtatt ctactttatg cgactaaaac
     4741 acgcgacaag aaaacgccag gaaaagggca gggcggcagc ctgtcgcgta acttaggact
     4801 tgtgcgacat gtcgttttca gaagaaggct gcactgaacg tcagaagccg actgcactat
     4861 agcagcggag gggttggatc aaagtacttt gatcccgagg ggaaccctgt ggttggcatg
     4921 cacatacaaa tggacgaacg gataaacctt ttcacgccct tttaaatatc cgattattct
     4981 aataaacgct cttttctctt aggtttaccc gccaatatat cctgtcaaac actgatagtt
     5041 taaaccagag aagacaacag agcggatccc tcaacttttg attcgctatt tgcagtgcac
     5101 ctgtggcgtt catcacatct tttgtgacac tgtttgcact ggtcattgct attacaaagg
     5161 accttcctga tgttgaagga gatcgaaagt aagtaactgc acgcataacc attttctttc
     5221 cgctctttgg ctcaatccat ttgacagtca aagacaatgt ttaaccagct ccgtttgata
     5281 tattgtcttt atgtgtttgt tcaagcatgt ttagttaatc atgcctttga ttgatcttga
     5341 ataggttcca aatatcaacc ctggcaacaa aacttggagt gagaaacatt gcattcctcg
     5401 gttctggact tctgctagta aattatgttt cagccatatc actagctttc tacatgcctc
     5461 aggtgaattc atctatttcc gtcttaacta tttcggttaa ttaaagcacg aacaccatta
     5521 ctgcatgtag aagcttgata aactatcgcc accaatttat ttttgttgcg atattgttac
     5581 tttcctcagt atgcagcttt gaaaagacca accctcttat cctttaacaa tgaacaggtt
     5641 tttagaggta gcttgatgat tcctgcacat gtgatcttgg cttcaggctt aattttccag
     5701 gtaaagcatt atgagatact cttatatctc ttacatactt ttgagataat gcacaagaac
     5761 ttcataacta tatgctttag tttctgcatt tgacactgcc aaattcatta atctctaata
     5821 tctttgttgt tgatctttgg tagacatggg tactagaaaa agcaaactac accaaggtaa
     5881 aatacttttg tacaaacata aactcgttat cacggaacat caatggagtg tatatctaac
     5941 ggagtgtaga aacatttgat tattgcagga agctatctca ggatattatc ggtttatatg
     6001 gaatctcttc tacgcagagt atctgttatt ccccttcctc tagctttcaa tttcatggtg
     6061 aggatatgca gttttctttg tatatcattc ttcttcttct ttgtagcttg gagtcaaaat
     6121 cggttccttc atgtacatac atcaaggata tgtccttctg aatttttata tcttgcaata
     6181 aaaatgcttg taccaattga aacaccagct ttttgagttc tatgatcact gacttggttc
     6241 taaccaaaaa aaaaaaatgt ttaatttaca tatctaaaag taggtttagg gaaacctaaa
     6301 cagtaaaata tttgtatatt attcgaattt cactcatcat aaaaacttaa attgcaccat
     6361 aaaattttgt tttactatta atgatgtaat ttgtgtaact taagataaaa ataatattcc
     6421 gtaagttaac cggctaaaac cacgtataaa ccagggaacc tgttaaaccg gttctttact
     6481 ggataaagaa atgaaagccc atgtagacag ctccattaga gcccaaaccc taaatttctc
     6541 atctatataa aaggagtgac attagggttt ttgttcgtcc tcttaaagct tctcgttttc
     6601 tctgccgtct ctctcattcg cgcgacgcaa acgatcttca ggtgatcttc tttctccaaa
     6661 tcctctctca taactctgat ttcgtacttg tgtatttgag ctcacgctct gtttctctca
     6721 ccacagcccc atggactata aggaccacga cggagactac aaggatcatg atattgatta
     6781 caaagacgat gacgataaga tggccccaaa gaagaagcgg aaggtcggta tccacggagt
     6841 cccagcagcc gacaagaagt acagcatcgg cctggacatc ggcaccaact ctgtgggctg
     6901 ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac
     6961 cgaccggcac agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac
     7021 agccgaggcc acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg
     7081 gatctgctat ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt
     7141 ccacagactg gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat
     7201 cttcggcaac atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct
     7261 gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct
     7321 ggcccacatg atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa
     7381 cagcgacgtg gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga
     7441 aaaccccatc aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa
     7501 gagcagacgg ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt
     7561 cggaaacctg attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct
     7621 ggccgaggat gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct
     7681 gctggcccag atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga
     7741 cgccatcctg ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag
     7801 cgcctctatg atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct
     7861 cgtgcggcag cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg
     7921 ctacgccggc tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc
     7981 catcctggaa aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct
     8041 gctgcggaag cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga
     8101 gctgcacgcc attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga
     8161 aaagatcgag aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg
     8221 aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt
     8281 cgaggaagtg gtggacaagg gcgcttccgc ccagagcttc atcgagcgga tgaccaactt
     8341 cgataagaac ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt
     8401 caccgtgtat aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc
     8461 cttcctgagc ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa
     8521 agtgaccgtg aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt
     8581 ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct
     8641 gaaaattatc aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga
     8701 tatcgtgctg accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac
     8761 ctatgcccac ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg
     8821 ctggggcagg ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac
     8881 aatcctggat ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca
     8941 cgacgacagc ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga
     9001 tagcctgcac gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct
     9061 gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa
     9121 catcgtgatc gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg
     9181 cgagagaatg aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga
     9241 acaccccgtg gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa
     9301 tgggcgggat atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt
     9361 ggaccatatc gtgcctcaga gctttctgaa ggacgactcc atcgacaaca aggtgctgac
     9421 cagaagcgac aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa
     9481 gatgaagaac tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga
     9541 caatctgacc aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa
     9601 gagacagctg gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg
     9661 gatgaacact aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct
     9721 gaagtccaag ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat
     9781 caacaactac caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat
     9841 caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt
     9901 gcggaagatg atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt
     9961 ctacagcaac atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg
    10021 gaagcggcct ctgatcgaga caaacggcga aaccggggag atcgtgtggg ataagggccg
    10081 ggattttgcc accgtgcgga aagtgctgag catgccccaa gtgaatatcg tgaaaaagac
    10141 cgaggtgcag acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgataa
    10201 gctgatcgcc agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac
    10261 cgtggcctat tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa
    10321 gagtgtgaaa gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc
    10381 catcgacttt ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct
    10441 gcctaagtac tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg
    10501 cgaactgcag aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct
    10561 ggccagccac tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt
    10621 tgtggaacag cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa
    10681 gagagtgatc ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg
    10741 ggataagccc atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct
    10801 gggagcccct gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag
    10861 caccaaagag gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac
    10921 acggatcgac ctgtctcagc tgggaggcga caaaaggccg gcggccacga aaaaggccgg
    10981 ccaggcaaaa aagaaaaagt aaaagggtgg gatatgaaga tgaagatgaa atatttggtg
    11041 tgtcaaataa aaagcttgtg tgcttaagtt tgtgtttttt tcttggcttg ttgtgttatg
    11101 aatttgtggc tttttctaat attaaatgaa tgtaagatct cattataatg aataaacaaa
    11161 tgtttctata atccattgtg aatgttttgt tggatctctt ctgcagcata taactactgt
    11221 atgtgctatg gtatggacta tggaatatga ttaaagataa gtctggaagc accgggcatc
    11281 ttcattctta agatatgaag ataatcttca aaaggcccct gggaatctga aagaagagaa
    11341 gcaggcccat ttatatggga aagaacaata gtatttctta tataggccca tttaagttga
    11401 aaacaatctt caaaagtccc acatcgctta gataagaaaa cgaagctgag tttatataca
    11461 gctagagtcg aagtagtgat tgagtccctt gctgttcaat ggttttagag ctagaaatag
    11521 caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt
    11581 tttttactgt cgagtaaagg gctaagacct aagctagcct agaatcgggc atcttcattc
    11641 ttaagatatg aagataatct tcaaaaggcc cctgggaatc tgaaagaaga gaagcaggcc
    11701 catttatatg ggaaagaaca atagtatttc ttatataggc ccatttaagt tgaaaacaat
    11761 cttcaaaagt cccacatcgc ttagataaga aaacgaagct gagtttatat acagctagag
    11821 tcgaagtagt gattgttgta cctcaggaac tgctgtttta gagctagaaa tagcaagtta
    11881 aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc ttttttttac
    11941 tgtcgagtat gaggtatgta ggtatagtaa catgaacaag aacagactca agtacaagat
    12001 cgcatacgaa aatggaaatg gcaatgtcac ttccacataa tcaaacacga atcctcatat
    12061 caacaaggcc tgagattcta actagctcat aacaacttag ccaatagtta cttgagacta
    12121 ccaaatgtat gtagaactaa agactaaggg acagagagtt cgtctaaaca ggtgaatcta
    12181 gtcgttgtta tctaataaac aattcagccc caaatgcaga acacacatag agctctctat
    12241 tgattcaaat tacgatctga tactgataac gtctagattt ttagggttaa agcaatcaat
    12301 cacctgacga ttcaaggtgg ttggatcatg acgattccag aaaacatcaa gcaagctctc
    12361 aaagctacac tctttgggat catactgaac tctaacaacc tcgttatgtc ccgtagtgcc
    12421 agtacagaca tcctcgtaac tcggattgtg cacgatgcca tggctatacc caacctcggt
    12481 cttggtcaca ccaggaactc tctggtaagc tagctccact ccccagaaac aaccggcgcc
    12541 aaattgcgcg aattgctgac ctgatgacgg aacatcatcg tcgggtcctt gggcgattgc
    12601 ggcggaagat gggtcagctt gggcttgagg acgagagccg aatccgagtc tgttgaaaag
    12661 gttgttcatt ggggatttgt atacggagat tggtcgtcga gaggtttgag ggaaaggaca
    12721 aatgggtttg gctctggaga aagagagtgc ggctttagag agagaattga gaggtttaga
    12781 gagagatgcg gcggcgatga gcgtgccgag gagagacgac gaggacctgc attatcaaag
    12841 cagtgacgtg gtgaaatttg gaacttttaa gaggcagata gatttattat ttgtatccat
    12901 tttcttcatt gttctagaat gtcgcggaac aaattttaaa actaaatcct aaatttttct
    12961 aattttgttg ccaatagtgg atatgtgggc cgtatagaag gaatctattg aaggctcaaa
    13021 cccatactga cgagcccaaa ggttcgtttt gcgttttatg tttcggttcg atgccaacgc
    13081 cacattctga gctaggcaaa aaacaaacgt gtctttgaat agactcctct cgttaacaca
    13141 tgcagcggct gcatggtgac gccattaaca cgtggcctac aattgcatga tgtctccatt
    13201 gacacgtgac ttctcgtctc ctttcttaat atatctaaca aacactccta cctcttccaa
    13261 aatatataca catctttttg atcaatctct cattcaaaat ctcattctct ctagtaaaca
    13321 agaacaaaaa aatggcggat acagctagag gaacccatca cgatatcatc ggcagggacc
    13381 agtacccgat gatgggccgt gaccgtgacc agtaccagat gtccggacga ggatctgact
    13441 actccaagtc taggcagatt gctaaagctg caactgctgt cacagctggt ggttccctcc
    13501 ttgttctctc cagccttacc cttgttggaa ctgtcatagc tttgactgtt gcaacacctc
    13561 tgctcgttat cttcagccca atccttgtcc cggctctcat cacagttgca ctcctcatca
    13621 ccggttttct ttcctctgga gggtttggca ttgccgctat aaccgttttc tcttggattt
    13681 acaagtaagc acacatttat catcttactt cataattttg tgcaatatgt gcatgcatgt
    13741 gttgagccag tagctttgga tcaatttttt tggtcgaata acaaatgtaa caataagaaa
    13801 ttgcaaattc tagggaacat ttggttaact aaatacgaaa tttgacctag ctagcttgaa
    13861 tgtgtctgtg tatatcatct atataggtaa aatgcttggt atgataccta ttgattgtga
    13921 ataggtacgc aacgggagag cacccacagg gatcagacaa gttggacagt gcaaggatga
    13981 agttgggaag caaagctcag gatctgaaag acagagctca gtactacgga cagcaacata
    14041 ctggtgggga acatgaccgt gaccgtactc gtggtggcca gcacactact atgagcgagc
    14101 tgattaagga gaacatgcac atgaagctgt acatggaggg caccgtgaac aaccaccact
    14161 tcaagtgcac atccgagggc gaaggcaagc cctacgaggg cacccagacc atgagaatca
    14221 aggtggtcga gggcggccct ctccccttcg ccttcgacat cctggctacc agcttcatgt
    14281 acggcagcag aaccttcatc aaccacaccc agggcatccc cgacttcttt aagcagtcct
    14341 tccctgaggg cttcacatgg gagagagtca ccacatacga ggacggaggt gtgctgaccg
    14401 ctactcagga caccagcctc caggacggct gcctcatcta caacgtcaag atcagagggg
    14461 tgaacttccc atccaacggc cctgtgatgc agaagaaaac actcggctgg gaagccaaca
    14521 ccgagatgct gtatccagct gacggaggac tggaaggcag aagcgacatg gccctgaagc
    14581 tcgtgggcgg gggccacctg atctgcaact tcaagaccac atacagatcc aagaaacccg
    14641 ctaagaacct caagatgccc ggcgtctact atgtggacca cagactggaa agaatcaagg
    14701 aggccgacaa agaaacctac gtcgagcagc acgaggtggc tgtggccaga tactgcgacc
    14761 tccctagcaa actggggcac aagtgattag agtttttcag atccgcgttt gtgttttctg
    14821 ggtttctcac ttaagcgtct gcgttttact tttgtattgg gtttggcgtt tagtagtttg
    14881 cggtagcgtt cttgttatgt gtaattacgc tttttcttct tgcttcagca gtttcggttg
    14941 aaatataaat cgaatcaagt ttcactttat cagcgttgtt ttaaattttg gcattaaatt
    15001 ggtgaaaatt gcttcaattt tgtatctaaa tagaagtgtc ctcattgtct tccgtaatcc
    15061 gtctaagcgt caatttgttt acaccacaat atatcctgcc accagccagc caacagctcc
    15121 ccgaccggca gctcggcaca aaatcaccac tcgatacagg cagcccatca gtccgggacg
    15181 gcgtcagcgg gagagccgtt gtaaggcggc agactttgct catgttaccg gcagaagatc
    15241 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
    15301 tggtcatgca tgatatatct cccaatttgt gtagggctta ttatgcacgc ttaaaaataa
    15361 taaaagcaga cttgacctga tagtttggct gtgagcaatt atgtgcttag tgcatctaac
    15421 gcttgagtta agccgcgccg cgaagcggcg tcggcttgaa cgaatttcta gctagacatt
    15481 atttgccgac taccttggtg atctcgcctt tcacgtagtg gacaaattct tccaactgat
    15541 ctgcgcgcga ggccaagcga tcttcttctt gtccaagata agcctgtcta gcttcaagta
    15601 tgacgggctg atactgggcc ggcaggcgct ccattgccca gtcggcagcg acatccttcg
    15661 gcgcgatttt gccggttact gcgctgtacc aaatgcggga caacgtaagc actacatttc
    15721 gctcatcgcc agcccagtcg ggcggcgagt tccatagcgt taaggtttca tttagcgcct
    15781 caaatagatc ctgttcagga accggatcaa agagttcctc cgccgctgga cctaccaagg
    15841 caacgctatg ttctcttgct tttgtcagca agatagccag atcaatgtcg atcgtggctg
    15901 gctcgaagat acctgcaaga atgtcattgc gctgccattc tccaaattgc agttcgcgct
    15961 tagctggata acgccacgga atgatgtcgt cgtgcacaac aatggtgact tctacagcgc
    16021 ggagaatctc gctctctcca ggggaagccg aagtttccaa aaggtcgttg atcaaagctc
    16081 gccgcgttgt ttcatcaagc cttacggtca ccgtaaccag caaatcaata tcactgtgtg
    16141 gcttcaggcc gccatccact gcggagccgt acaaatgtac ggccagcaac gtcggttcga
    16201 gatggcgctc gatgacgcca actacctctg atagttgagt cgatacttcg gcgatcaccg
    16261 cttcccccat gatgtttaac tttgttttag ggcgactgcc ctgctgcgta acatcgttgc
    16321 tgctccataa catcaaacat cgacccacgg cgtaacgcgc ttgctgcttg gatgcccgag
    16381 gcatagactg taccccaaaa aaacagtcat aacaagccat gaaaaccgcc actgcgccgt
    16441 taccaccgct gcgttcggtc aaggttctgg accagttgcg tgagcgcata cgctacttgc
    16501 attacagctt acgaaccgaa caggcttatg tccactgggt tcgtgcccga attgatcaca
    16561 ggcagcaacg ctctgtcatc gttacaatca acatgctacc ctccgcgaga tcatccgtgt
    16621 ttcaaacccg gcagcttagt
//
