LOCUS       sequence-175593-       14215 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cas9 only expression, Cas9 Type: AtCas9, Plant Selection: 2x35S:hpt 
            II.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14215)
  AUTHORS   Cermak T, Curtin SJ, Gil-Humanes J, Cegan R, Kono TJY, Konecna E, 
            Belanto JJ, Starker CG, Mathre JW, Greenstein RL, Voytas DF
  TITLE     A multi-purpose toolkit to enable advanced genome engineering in 
            plants.
  JOURNAL   Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 
            10.1105/tpc.16.00922.
  PUBMED    28522548
REFERENCE   2  (bases 1 to 14215)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant Cell.
            2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922."
COMMENT     Sequence Label: pAtCas9_1
FEATURES             Location/Qualifiers
     source          1..14215
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(291..1085)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    1510..1534
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    1612..1786
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(1826..2851)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     promoter        complement(2919..3595)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     protein_bind    3786..3807
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3822..3827,3828..3845,3846..3852)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3860..3876
                     /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)."
     primer_bind     3865..3887
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3884..3900
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3884..3900
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        4431..4776
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             4810..8913
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             4810..8913
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             8926..8946
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      8957..9205
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     primer_bind     complement(9227..9244)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9227..9243)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9236..9258)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    9446..9470
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             10771..11400
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             11834..12901
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     rep_origin      12967..13161
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(13397..13419)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    13505..13645
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     13560..13579
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(13660..13677)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(13831..14215,1..204))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13911..13930)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
       61 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
      121 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
      181 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
      241 cagtggaacg aaaactcacg ttaagggatt ttggtcatgc attctaggta ctaaaacaat
      301 tcatccagta aaatataata ttttattttc tcccaatcag gcttgatccc cagtaagtca
      361 aaaaatagct cgacatactg ttcttccccg atatcctccc tgatcgaccg gacgcagaag
      421 gcaatgtcat accacttgtc cgccctgccg cttctcccaa gatcaataaa gccacttact
      481 ttgccatctt tcacaaagat gttgctgtct cccaggtcgc cgtgggaaaa gacaagttcc
      541 tcttcgggct tttccgtctt taaaaaatca tacagctcgc gcggatcttt aaatggagtg
      601 tcttcttccc agttttcgca atccacatcg gccagatcgt tattcagtaa gtaatccaat
      661 tcggctaagc ggctgtctaa gctattcgta tagggacaat ccgatatgtc gatggagtga
      721 aagagcctga tgcactccgc atacagctcg ataatctttt cagggctttg ttcatcttca
      781 tactcttccg agcaaaggac gccatcggcc tcactcatga gcagattgct ccagccatca
      841 tgccgttcaa agtgcaggac ctttggaaca ggcagctttc cttccagcca tagcatcatg
      901 tccttttccc gttccacatc ataggtggtc cctttatacc ggctgtccgt catttttaaa
      961 tataggtttt cattttctcc caccagctta tataccttag caggagacat tccttccgta
     1021 tcttttacgc agcggtattt ttcgatcagt tttttcaatt ccggtgatat tctcatttta
     1081 gccatttatt atttccttcc tcttttctac agtatttaaa gataccccaa gaagctaatt
     1141 ataacaagac gaactccaat tcactgttcc ttgcattcta aaaccttaaa taccagaaaa
     1201 cagctttttc aaagttgttt tcaaagttgg cgtataacat agtatcgacg gagccgattt
     1261 tgaaaccgcg gtgatcacag gcagcaacgc tctgtcatcg ttacaatcaa catgctaccc
     1321 tccgcgagat catccgtgtt tcaaacccgg cagcttagtt gccgttcttc cgaatagcat
     1381 cggtaacatg agcaaagtct gccgccttac aacggctctc ccgctgacgc cgtcccggac
     1441 tgatgggctg cctgtatcga gtggtgattt tgtgccgagc tgccggtcgg ggagctgttg
     1501 gctggctggt ggcaggatat attgtggtgt aaacaaattg acgcttagac aacttaataa
     1561 cacattgcgg acgtttttaa tgtactgaat taacgccgaa ttaattcggg ggatctggat
     1621 tttagtactg gattttggtt ttaggaatta gaaattttat tgatagaagt attttacaaa
     1681 tacaaataca tactaagggt ttcttatatg ctcaacacat gagcgaaacc ctataggaac
     1741 cctaattccc ttatctggga actactcaca cattattatg gagaaactcg agcttgtcga
     1801 tcgacagatc cggtcggcat ctactctatt tctttgccct cggacgagtg ctggggcgtc
     1861 ggtttccact atcggcgagt acttctacac agccatcggt ccagacggcc gcgcttctgc
     1921 gggcgatttg tgtacgcccg acagtcccgg ctccggatcg gacgattgcg tcgcatcgac
     1981 cctgcgccca agctgcatca tcgaaattgc cgtcaaccaa gctctgatag agttggtcaa
     2041 gaccaatgcg gagcatatac gcccggagtc gtggcgatcc tgcaagctcc ggatgcctcc
     2101 gctcgaagta gcgcgtctgc tgctccatac aagccaacca cggcctccag aagaagatgt
     2161 tggcgacctc gtattgggaa tccccgaaca tcgcctcgct ccagtcaatg accgctgtta
     2221 tgcggccatt gtccgtcagg acattgttgg agccgaaatc cgcgtgcacg aggtgccgga
     2281 cttcggggca gtcctcggcc caaagcatca gctcatcgag agcctgcgcg acggacgcac
     2341 tgacggtgtc gtccatcaca gtttgccagt gatacacatg gggatcagca atcgcgcata
     2401 tgaaatcacg ccatgtagtg tattgaccga ttccttgcgg tccgaatggg ccgaacccgc
     2461 tcgtctggct aagatcggcc gcagcgatcg catccatagc ctccgcgacc ggttgtagaa
     2521 cagcgggcag ttcggtttca ggcaggtctt gcaacgtgac accctgtgca cggcgggaga
     2581 tgcaataggt caggctctcg ctaaactccc caatgtcaag cacttccgga atcgggagcg
     2641 cggccgatgc aaagtgccga taaacataac gatctttgta gaaaccatcg gcgcagctat
     2701 ttacccgcag gacatatcca cgccctccta catcgaagct gaaagcacga gattcttcgc
     2761 cctccgagag ctgcatcagg tcggagacgc tgtcgaactt ttcgatcaga aacttctcga
     2821 cagacgtcgc ggtgagttca ggctttttca tatctcattg cccccccgga tctgcgaaag
     2881 ctcgagagag atagatttgt agagagagac tggtgatttc agcgtgtcct ctccaaatga
     2941 aatgaacttc cttatataga ggaagggtct tgcgaaggat agtgggattg tgcgtcatcc
     3001 cttacgtcag tggagatatc acatcaatcc acttgctttg aagacgtggt tggaacgtct
     3061 tctttttcca cgatgctcct cgtgggtggg ggtccatctt tgggaccact gtcggcagag
     3121 gcatcttgaa cgatagcctt tcctttatcg caatgatggc atttgtaggt gccaccttcc
     3181 ttttctactg tccttttgat gaagtgacag atagctgggc aatggaatcc gaggaggttt
     3241 cccgatatta ccctttgttg aaaagtctca atagcccttt ggtcttctga gactgtatct
     3301 ttgatattct tggagtagac gagagtgtcg tgctccacca tgttcacatc aatccacttg
     3361 ctttgaagac gtggttggaa cgtcttcttt ttccacgatg ctcctcgtgg gtgggggtcc
     3421 atctttggga ccactgtcgg cagaggcatc ttgaacgata gcctttcctt tatcgcaatg
     3481 atggcatttg taggtgccac cttccttttc tactgtcctt ttgatgaagt gacagatagc
     3541 tgggcaatgg aatccgagga ggtttcccga tattaccctt tgttgaaaag tctcaatagc
     3601 cctttggtct tctgagactg tatctttgat attcttggag tagacgagag tgtcgtgctc
     3661 caccatgttg gcaagctgct ctagccaata cgcaaaccgc ctctccccgc gcgttggccg
     3721 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     3781 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     3841 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     3901 catgattacg aattcgagct cggtacccgg ggatcggcgc gccagatttg ccttttcaat
     3961 ttcagaaaga atgctaaccc acagatggtt agagaggctt acgcagcagg tatcatcaag
     4021 acgatctacc cgagcaataa tctccaggaa atcaaatacc ttcccaagaa ggttaaagat
     4081 gcagtcaaaa gattcaggac taactgcatc aagaacacag agaaagatat atttctcaag
     4141 atcagaagta ctattccagt atggacgatt caaggcttgc ttcacaaacc aaggcaagta
     4201 atagagattg gagtctctaa aaaggtagtt cccactgaat caaaggccat ggagtcaaag
     4261 attcaaatag aggacctaac agaactcgcc gtaaagactg gcgaacagtt catacagagt
     4321 ctcttacgac tcaatgacaa gaagaaaatc ttcgtcaaca tggtggagca cgacacactt
     4381 gtctactcca aaaatatcaa agatacagtc tcagaagacc aaagggcaat tgagactttt
     4441 caacaaaggg taatatccgg aaacctcctc ggattccatt gcccagctat ctgtcacttt
     4501 attgtgaaga tagtggaaaa ggaaggtggc tcctacaaat gccatcattg cgataaagga
     4561 aaggccatcg ttgaagatgc ctctgccgac agtggtccca aagatggacc cccacccacg
     4621 aggagcatcg tggaaaaaga agacgttcca accacgtctt caaagcaagt ggattgatgt
     4681 gatatctcca ctgacgtaag ggatgacgca caatcccact atccttcgca agacccttcc
     4741 tctatataag gaagttcatt tcatttggag agaacacggg ggactcctgc aggtagatcg
     4801 ctcgtcgaca tggataagaa gtactctatc ggactcgata tcggaactaa ctctgtggga
     4861 tgggctgtga tcaccgatga gtacaaggtg ccatctaaga agttcaaggt tctcggaaac
     4921 accgataggc actctatcaa gaaaaacctt atcggtgctc tcctcttcga ttctggtgaa
     4981 actgctgagg ctaccagact caagagaacc gctagaagaa ggtacaccag aagaaagaac
     5041 aggatctgct acctccaaga gatcttctct aacgagatgg ctaaagtgga tgattcattc
     5101 ttccacaggc tcgaagagtc attcctcgtg gaagaagata agaagcacga gaggcaccct
     5161 atcttcggaa acatcgttga tgaggtggca taccacgaga agtaccctac tatctaccac
     5221 ctcagaaaga agctcgttga ttctactgat aaggctgatc tcaggctcat ctacctcgct
     5281 ctcgctcaca tgatcaagtt cagaggacac ttcctcatcg agggtgatct caaccctgat
     5341 aactctgatg tggataagtt gttcatccag ctcgtgcaga cctacaacca gcttttcgaa
     5401 gagaacccta tcaacgcttc aggtgtggat gctaaggcta tcctctctgc taggctctct
     5461 aagtcaagaa ggcttgagaa cctcattgct cagctccctg gtgagaagaa gaacggactt
     5521 ttcggaaact tgatcgctct ctctctcgga ctcaccccta acttcaagtc taacttcgat
     5581 ctcgctgagg atgcaaagct ccagctctca aaggatacct acgatgatga tctcgataac
     5641 ctcctcgctc agatcggaga tcagtacgct gatttgttcc tcgctgctaa gaacctctct
     5701 gatgctatcc tcctcagtga tatcctcaga gtgaacaccg agatcaccaa ggctccactc
     5761 tcagcttcta tgatcaagag atacgatgag caccaccagg atctcacact tctcaaggct
     5821 cttgttagac agcagctccc agagaagtac aaagagattt tcttcgatca gtctaagaac
     5881 ggatacgctg gttacatcga tggtggtgca tctcaagaag agttctacaa gttcatcaag
     5941 cctatcctcg agaagatgga tggaaccgag gaactcctcg tgaagctcaa tagagaggat
     6001 cttctcagaa agcagaggac cttcgataac ggatctatcc ctcatcagat ccacctcgga
     6061 gagttgcacg ctatccttag aaggcaagag gatttctacc cattcctcaa ggataacagg
     6121 gaaaagattg agaagattct caccttcaga atcccttact acgtgggacc tctcgctaga
     6181 ggaaactcaa gattcgcttg gatgaccaga aagtctgagg aaaccatcac cccttggaac
     6241 ttcgaagagg tggtggataa gggtgctagt gctcagtctt tcatcgagag gatgaccaac
     6301 ttcgataaga accttccaaa cgagaaggtg ctccctaagc actctttgct ctacgagtac
     6361 ttcaccgtgt acaacgagtt gaccaaggtt aagtacgtga ccgagggaat gaggaagcct
     6421 gcttttttgt caggtgagca aaagaaggct atcgttgatc tcttgttcaa gaccaacaga
     6481 aaggtgaccg tgaagcagct caaagaggat tacttcaaga aaatcgagtg cttcgattca
     6541 gttgagattt ctggtgttga ggataggttc aacgcatctc tcggaaccta ccacgatctc
     6601 ctcaagatca ttaaggataa ggatttcttg gataacgagg aaaacgagga tatcttggag
     6661 gatatcgttc ttaccctcac cctctttgaa gatagagaga tgattgaaga aaggctcaag
     6721 acctacgctc atctcttcga tgataaggtg atgaagcagt tgaagagaag aagatacact
     6781 ggttggggaa ggctctcaag aaagctcatt aacggaatca gggataagca gtctggaaag
     6841 acaatccttg atttcctcaa gtctgatgga ttcgctaaca gaaacttcat gcagctcatc
     6901 cacgatgatt ctctcacctt taaagaggat atccagaagg ctcaggtttc aggacagggt
     6961 gatagtctcc atgagcatat cgctaacctc gctggatctc ctgcaatcaa gaagggaatc
     7021 ctccagactg tgaaggttgt ggatgagttg gtgaaggtga tgggaaggca taagcctgag
     7081 aacatcgtga tcgaaatggc tagagagaac cagaccactc agaagggaca gaagaactct
     7141 agggaaagga tgaagaggat cgaggaaggt atcaaagagc ttggatctca gatcctcaaa
     7201 gagcaccctg ttgagaacac tcagctccag aatgagaagc tctacctcta ctacctccag
     7261 aacggaaggg atatgtatgt ggatcaagag ttggatatca acaggctctc tgattacgat
     7321 gttgatcata tcgtgccaca gtcattcttg aaggatgatt ctatcgataa caaggtgctc
     7381 accaggtctg ataagaacag gggtaagagt gataacgtgc caagtgaaga ggttgtgaag
     7441 aaaatgaaga actattggag gcagctcctc aacgctaagc tcatcactca gagaaagttc
     7501 gataacttga ctaaggctga gaggggagga ctctctgaat tggataaggc aggattcatc
     7561 aagaggcagc ttgtggaaac caggcagatc actaagcacg ttgcacagat cctcgattct
     7621 aggatgaaca ccaagtacga tgagaacgat aagttgatca gggaagtgaa ggttatcacc
     7681 ctcaagtcaa agctcgtgtc tgatttcaga aaggatttcc aattctacaa ggtgagggaa
     7741 atcaacaact accaccacgc tcacgatgct taccttaacg ctgttgttgg aaccgctctc
     7801 atcaagaagt atcctaagct cgagtcagag ttcgtgtacg gtgattacaa ggtgtacgat
     7861 gtgaggaaga tgatcgctaa gtctgagcaa gagatcggaa aggctaccgc taagtatttc
     7921 ttctactcta acatcatgaa tttcttcaag accgagatta ccctcgctaa cggtgagatc
     7981 agaaagaggc cactcatcga gacaaacggt gaaacaggtg agatcgtgtg ggataaggga
     8041 agggatttcg ctaccgttag aaaggtgctc tctatgccac aggtgaacat cgttaagaaa
     8101 accgaggtgc agaccggtgg attctctaaa gagtctatcc tccctaagag gaactctgat
     8161 aagctcattg ctaggaagaa ggattgggac cctaagaaat acggtggttt cgattctcct
     8221 accgtggctt actctgttct cgttgtggct aaggttgaga agggaaagag taagaagctc
     8281 aagtctgtta aggaacttct cggaatcact atcatggaaa ggtcatcttt cgagaagaac
     8341 ccaatcgatt tcctcgaggc taagggatac aaagaggtta agaaggatct catcatcaag
     8401 ctcccaaagt actcactctt cgaactcgag aacggtagaa agaggatgct cgcttctgct
     8461 ggtgagcttc aaaagggaaa cgagcttgct ctcccatcta agtacgttaa ctttctttac
     8521 ctcgcttctc actacgagaa gttgaaggga tctccagaag ataacgagca gaagcaactt
     8581 ttcgttgagc agcacaagca ctacttggat gagatcatcg agcagatctc tgagttctct
     8641 aaaagggtga tcctcgctga tgcaaacctc gataaggtgt tgtctgctta caacaagcac
     8701 agagataagc ctatcaggga acaggcagag aacatcatcc atctcttcac ccttaccaac
     8761 ctcggtgctc ctgctgcttt caagtacttc gatacaacca tcgataggaa gagatacacc
     8821 tctaccaaag aagtgctcga tgctaccctc atccatcagt ctatcactgg actctacgag
     8881 actaggatcg atctctcaca gctcggtggt gattcaaggg ctgatcctaa gaagaagagg
     8941 aaggtttgac tcgagatatg aagatgaaga tgaaatattt ggtgtgtcaa ataaaaagct
     9001 tgtgtgctta agtttgtgtt tttttcttgg cttgttgtgt tatgaatttg tggctttttc
     9061 taatattaaa tgaatgtaag atcacattat aatgaataaa caaatgtttc tataatccat
     9121 tgtgaatgtt ttgttggatc tcttctgcag catataacta ctgtatgtgc tatggtatgg
     9181 actatggaat atgattaaag ataaggagct ccggtgacgg actggcactg gccgtcgttt
     9241 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     9301 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     9361 tgcgcagcct gaatggcgaa tgctagagca gcttgagctt ggatcagatt gtcgtttccc
     9421 gccttcagtt taaactatca gtgtttgaca ggatatattg gcgggtaaac ctaagagaaa
     9481 agagcgttta ttagaataat cggatattta aaagggcgtg aaaaggttta tccgttcgtc
     9541 catttgtatg tgcatgccaa ccacagggtt cccctcggga tcaaagtact ttgatccaac
     9601 ccctccgctg ctatagtgca gtcggcttct gacgttcagt gcagccgtct tctgaaaacg
     9661 acatgtcgca caagtcctaa gttacgcgac aggctgccgc cctgcccttt tcctggcgtt
     9721 ttcttgtcgc gtgttttagt cgcataaagt agaatacttg cgactagaac cggagacatt
     9781 acgccatgaa caagagcgcc gccgctggcc tgctgggcta tgcccgcgtc agcaccgacg
     9841 accaggactt gaccaaccaa cgggccgaac tgcacgcggc cggctgcacc aagctgtttt
     9901 ccgagaagat caccggcacc aggcgcgacc gcccggagct ggccaggatg cttgaccacc
     9961 tacgccctgg cgacgttgtg acagtgacca ggctagaccg cctggcccgc agcacccgcg
    10021 acctactgga cattgccgag cgcatccagg aggccggcgc gggcctgcgt agcctggcag
    10081 agccgtgggc cgacaccacc acgccggccg gccgcatggt gttgaccgtg ttcgccggca
    10141 ttgccgagtt cgagcgttcc ctaatcatcg accgcacccg gagcgggcgc gaggccgcca
    10201 aggcccgagg cgtgaagttt ggcccccgcc ctaccctcac cccggcacag atcgcgcacg
    10261 cccgcgagct gatcgaccag gaaggccgca ccgtgaaaga ggcggctgca ctgcttggcg
    10321 tgcatcgctc gaccctgtac cgcgcacttg agcgcagcga ggaagtgacg cccaccgagg
    10381 ccaggcggcg cggtgccttc cgtgaggacg cattgaccga ggccgacgcc ctggcggccg
    10441 ccgagaatga acgccaagag gaacaagcat gaaaccgcac caggacggcc aggacgaacc
    10501 gtttttcatt accgaagaga tcgaggcgga gatgatcgcg gccgggtacg tgttcgagcc
    10561 gcccgcgcac gtctcaaccg tgcggctgca tgaaatcctg gccggtttgt ctgatgccaa
    10621 gctggcggcc tggccggcca gcttggccgc tgaagaaacc gagcgccgcc gtctaaaaag
    10681 gtgatgtgta tttgagtaaa acagcttgcg tcatgcggtc gctgcgtata tgatgcgatg
    10741 agtaaataaa caaatacgca aggggaacgc atgaaggtta tcgctgtact taaccagaaa
    10801 ggcgggtcag gcaagacgac catcgcaacc catctagccc gcgccctgca actcgccggg
    10861 gccgatgttc tgttagtcga ttccgatccc cagggcagtg cccgcgattg ggcggccgtg
    10921 cgggaagatc aaccgctaac cgttgtcggc atcgaccgcc cgacgattga ccgcgacgtg
    10981 aaggccatcg gccggcgcga cttcgtagtg atcgacggag cgccccaggc ggcggacttg
    11041 gctgtgtccg cgatcaaggc agccgacttc gtgctgattc cggtgcagcc aagcccttac
    11101 gacatatggg ccaccgccga cctggtggag ctggttaagc agcgcattga ggtcacggat
    11161 ggaaggctac aagcggcctt tgtcgtgtcg cgggcgatca aaggcacgcg catcggcggt
    11221 gaggttgccg aggcgctggc cgggtacgag ctgcccattc ttgagtcccg tatcacgcag
    11281 cgcgtgagct acccaggcac tgccgccgcc ggcacaaccg ttcttgaatc agaacccgag
    11341 ggcgacgctg cccgcgaggt ccaggcgctg gccgctgaaa ttaaatcaaa actcatttga
    11401 gttaatgagg taaagagaaa atgagcaaaa gcacaaacac gctaagtgcc ggccgtccga
    11461 gcgcacgcag cagcaaggct gcaacgttgg ccagcctggc agacacgcca gccatgaagc
    11521 gggtcaactt tcagttgccg gcggaggatc acaccaagct gaagatgtac gcggtacgcc
    11581 aaggcaagac cattaccgag ctgctatctg aatacatcgc gcagctacca gagtaaatga
    11641 gcaaatgaat aaatgagtag atgaatttta gcggctaaag gaggcggcat ggaaaatcaa
    11701 gaacaaccag gcaccgacgc cgtggaatgc cccatgtgtg gaggaacggg cggttggcca
    11761 ggcgtaagcg gctgggttgt ctgccggccc tgcaatggca ctggaacccc caagcccgag
    11821 gaatcggcgt gacggtcgca aaccatccgg cccggtacaa atcggcgcgg cgctgggtga
    11881 tgacctggtg gagaagttga aggccgcgca ggccgcccag cggcaacgca tcgaggcaga
    11941 agcacgcccc ggtgaatcgt ggcaagcggc cgctgatcga atccgcaaag aatcccggca
    12001 accgccggca gccggtgcgc cgtcgattag gaagccgccc aagggcgacg agcaaccaga
    12061 ttttttcgtt ccgatgctct atgacgtggg cacccgcgat agtcgcagca tcatggacgt
    12121 ggccgttttc cgtctgtcga agcgtgaccg acgagctggc gaggtgatcc gctacgagct
    12181 tccagacggg cacgtagagg tttccgcagg gccggccggc atggccagtg tgtgggatta
    12241 cgacctggta ctgatggcgg tttcccatct aaccgaatcc atgaaccgat accgggaagg
    12301 gaagggagac aagcccggcc gcgtgttccg tccacacgtt gcggacgtac tcaagttctg
    12361 ccggcgagcc gatggcggaa agcagaaaga cgacctggta gaaacctgca ttcggttaaa
    12421 caccacgcac gttgccatgc agcgtacgaa gaaggccaag aacggccgcc tggtgacggt
    12481 atccgagggt gaagccttga ttagccgcta caagatcgta aagagcgaaa ccgggcggcc
    12541 ggagtacatc gagatcgagc tagctgattg gatgtaccgc gagatcacag aaggcaagaa
    12601 cccggacgtg ctgacggttc accccgatta ctttttgatc gatcccggca tcggccgttt
    12661 tctctaccgc ctggcacgcc gcgccgcagg caaggcagaa gccagatggt tgttcaagac
    12721 gatctacgaa cgcagtggca gcgccggaga gttcaagaag ttctgtttca ccgtgcgcaa
    12781 gctgatcggg tcaaatgacc tgccggagta cgatttgaag gaggaggcgg ggcaggctgg
    12841 cccgatccta gtcatgcgct accgcaacct gatcgagggc gaagcatccg ccggttccta
    12901 atgtacggag cagatgctag ggcaaattgc cctagcaggg gaaaaaggtc gaaaaggtct
    12961 ctttcctgtg gatagcacgt acattgggaa cccaaagccg tacattggga accggaaccc
    13021 gtacattggg aacccaaagc cgtacattgg gaaccggtca cacatgtaag tgactgatat
    13081 aaaagagaaa aaaggcgatt tttccgccta aaactcttta aaacttatta aaactcttaa
    13141 aacccgcctg gcctgtgcat aactgtctgg ccagcgcaca gccgaagagc tgcaaaaagc
    13201 gcctaccctt cggtcgctgc gctccctacg ccccgccgct tcgcgtcggc ctatcgcggc
    13261 cgctggccgc tcaaaaatgg ctggcctacg gccaggcaat ctaccagggc gcggacaagc
    13321 cgcgccgtcg ccactcgacc gccggcgccc acatcaaggc accctgcctc gcgcgtttcg
    13381 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
    13441 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
    13501 ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc
    13561 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
    13621 cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg
    13681 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
    13741 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
    13801 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
    13861 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
    13921 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
    13981 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
    14041 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
    14101 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
    14161 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtat
//
