LOCUS       sequence-182265-       14676 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Direct Cloning, Type: T-DNA, Engineering Reagent: 35S:AtCas9 + 
            AtU6:gRNA, Plant Selection: 2x35S:bar.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14676)
  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 14676)
  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: pDIRECT_23A
FEATURES             Location/Qualifiers
     source          1..14676
                     /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(1793..2344)
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     promoter        complement(2389..3065)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     protein_bind    3256..3277
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3292..3297,3298..3315,3316..3322)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3330..3346
                     /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     3354..3370
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3900..4245
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             4279..8382
                     /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             4279..8382
                     /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             8395..8415
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      8426..8674
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     CDS             complement(9141..9446)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGTCL"
     primer_bind     9311..9327
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9337..9356
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        9337..9355
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9403..9423)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(9403..9421)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(9442..9458)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    9466..9482
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(9490..9496,9497..9514,9515..9520))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9535..9556
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_RNA        9595..9670
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(9687..9703)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    9906..9930
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             11231..11860
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             12289..13362
                     /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"
     rep_origin      13428..13622
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(13858..13880)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    13966..14106
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     14021..14040
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(14121..14138)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(14292..14676,1..204))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(14372..14391)
                     /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 agtcaaatct
     1801 cggtgacggg caggaccgga cggggcggta ccggcaggct gaagtccagc tgccagaaac
     1861 ccacgtcatg ccagttcccg tgcttgaagc cggccgcccg cagcatgccg cggggggcat
     1921 atccgagcgc ctcgtgcatg cgcacgctcg ggtcgttggg cagcccgatg acagcgacca
     1981 cgctcttgaa gccctgtgcc tccagggact tcagcagatg ggtgtagagc gtggagccca
     2041 gtcccgtccg ctggtggcgg ggggagacgt acacggtcga ctcggccgtc cagtcgtagg
     2101 cgttgcgtgc cttccagggg cccgcgtagg cgatgccggc gacctcgccg tccacctcgg
     2161 cgacgagcca gggatagcgc tcccgcagac ggacgaggtc gtccgtccac tcctgcggtt
     2221 cctgcggctc ggtacggaag ttgaccgtgc ttgtctcgat gtagtggttg acgatggtgc
     2281 agaccgccgg catgtccgcc tcggtggcac ggcggatgtc ggccgggcgt cgttctgggc
     2341 tcatggtaga ctcgagagag atagatttgt agagagagac tggtgatttc agcgtgtcct
     2401 ctccaaatga aatgaacttc cttatataga ggaagggtct tgcgaaggat agtgggattg
     2461 tgcgtcatcc cttacgtcag tggagatatc acatcaatcc acttgctttg aagacgtggt
     2521 tggaacgtct tctttttcca cgatgctcct cgtgggtggg ggtccatctt tgggaccact
     2581 gtcggcagag gcatcttgaa cgatagcctt tcctttatcg caatgatggc atttgtaggt
     2641 gccaccttcc ttttctactg tccttttgat gaagtgacag atagctgggc aatggaatcc
     2701 gaggaggttt cccgatatta ccctttgttg aaaagtctca atagcccttt ggtcttctga
     2761 gactgtatct ttgatattct tggagtagac gagagtgtcg tgctccacca tgttcacatc
     2821 aatccacttg ctttgaagac gtggttggaa cgtcttcttt ttccacgatg ctcctcgtgg
     2881 gtgggggtcc atctttggga ccactgtcgg cagaggcatc ttgaacgata gcctttcctt
     2941 tatcgcaatg atggcatttg taggtgccac cttccttttc tactgtcctt ttgatgaagt
     3001 gacagatagc tgggcaatgg aatccgagga ggtttcccga tattaccctt tgttgaaaag
     3061 tctcaatagc cctttggtct tctgagactg tatctttgat attcttggag tagacgagag
     3121 tgtcgtgctc caccatgttg gcaagctgct ctagccaata cgcaaaccgc ctctccccgc
     3181 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     3241 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     3301 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     3361 cagctatgac atgattacga attcgagctc ggtacccggg gatcggcgcg ccagatttgc
     3421 cttttcaatt tcagaaagaa tgctaaccca cagatggtta gagaggctta cgcagcaggt
     3481 atcatcaaga cgatctaccc gagcaataat ctccaggaaa tcaaatacct tcccaagaag
     3541 gttaaagatg cagtcaaaag attcaggact aactgcatca agaacacaga gaaagatata
     3601 tttctcaaga tcagaagtac tattccagta tggacgattc aaggcttgct tcacaaacca
     3661 aggcaagtaa tagagattgg agtctctaaa aaggtagttc ccactgaatc aaaggccatg
     3721 gagtcaaaga ttcaaataga ggacctaaca gaactcgccg taaagactgg cgaacagttc
     3781 atacagagtc tcttacgact caatgacaag aagaaaatct tcgtcaacat ggtggagcac
     3841 gacacacttg tctactccaa aaatatcaaa gatacagtct cagaagacca aagggcaatt
     3901 gagacttttc aacaaagggt aatatccgga aacctcctcg gattccattg cccagctatc
     3961 tgtcacttta ttgtgaagat agtggaaaag gaaggtggct cctacaaatg ccatcattgc
     4021 gataaaggaa aggccatcgt tgaagatgcc tctgccgaca gtggtcccaa agatggaccc
     4081 ccacccacga ggagcatcgt ggaaaaagaa gacgttccaa ccacgtcttc aaagcaagtg
     4141 gattgatgtg atatctccac tgacgtaagg gatgacgcac aatcccacta tccttcgcaa
     4201 gacccttcct ctatataagg aagttcattt catttggaga gaacacgggg gactcctgca
     4261 ggtagatcgc tcgtcgacat ggataagaag tactctatcg gactcgatat cggaactaac
     4321 tctgtgggat gggctgtgat caccgatgag tacaaggtgc catctaagaa gttcaaggtt
     4381 ctcggaaaca ccgataggca ctctatcaag aaaaacctta tcggtgctct cctcttcgat
     4441 tctggtgaaa ctgctgaggc taccagactc aagagaaccg ctagaagaag gtacaccaga
     4501 agaaagaaca ggatctgcta cctccaagag atcttctcta acgagatggc taaagtggat
     4561 gattcattct tccacaggct cgaagagtca ttcctcgtgg aagaagataa gaagcacgag
     4621 aggcacccta tcttcggaaa catcgttgat gaggtggcat accacgagaa gtaccctact
     4681 atctaccacc tcagaaagaa gctcgttgat tctactgata aggctgatct caggctcatc
     4741 tacctcgctc tcgctcacat gatcaagttc agaggacact tcctcatcga gggtgatctc
     4801 aaccctgata actctgatgt ggataagttg ttcatccagc tcgtgcagac ctacaaccag
     4861 cttttcgaag agaaccctat caacgcttca ggtgtggatg ctaaggctat cctctctgct
     4921 aggctctcta agtcaagaag gcttgagaac ctcattgctc agctccctgg tgagaagaag
     4981 aacggacttt tcggaaactt gatcgctctc tctctcggac tcacccctaa cttcaagtct
     5041 aacttcgatc tcgctgagga tgcaaagctc cagctctcaa aggataccta cgatgatgat
     5101 ctcgataacc tcctcgctca gatcggagat cagtacgctg atttgttcct cgctgctaag
     5161 aacctctctg atgctatcct cctcagtgat atcctcagag tgaacaccga gatcaccaag
     5221 gctccactct cagcttctat gatcaagaga tacgatgagc accaccagga tctcacactt
     5281 ctcaaggctc ttgttagaca gcagctccca gagaagtaca aagagatttt cttcgatcag
     5341 tctaagaacg gatacgctgg ttacatcgat ggtggtgcat ctcaagaaga gttctacaag
     5401 ttcatcaagc ctatcctcga gaagatggat ggaaccgagg aactcctcgt gaagctcaat
     5461 agagaggatc ttctcagaaa gcagaggacc ttcgataacg gatctatccc tcatcagatc
     5521 cacctcggag agttgcacgc tatccttaga aggcaagagg atttctaccc attcctcaag
     5581 gataacaggg aaaagattga gaagattctc accttcagaa tcccttacta cgtgggacct
     5641 ctcgctagag gaaactcaag attcgcttgg atgaccagaa agtctgagga aaccatcacc
     5701 ccttggaact tcgaagaggt ggtggataag ggtgctagtg ctcagtcttt catcgagagg
     5761 atgaccaact tcgataagaa ccttccaaac gagaaggtgc tccctaagca ctctttgctc
     5821 tacgagtact tcaccgtgta caacgagttg accaaggtta agtacgtgac cgagggaatg
     5881 aggaagcctg cttttttgtc aggtgagcaa aagaaggcta tcgttgatct cttgttcaag
     5941 accaacagaa aggtgaccgt gaagcagctc aaagaggatt acttcaagaa aatcgagtgc
     6001 ttcgattcag ttgagatttc tggtgttgag gataggttca acgcatctct cggaacctac
     6061 cacgatctcc tcaagatcat taaggataag gatttcttgg ataacgagga aaacgaggat
     6121 atcttggagg atatcgttct taccctcacc ctctttgaag atagagagat gattgaagaa
     6181 aggctcaaga cctacgctca tctcttcgat gataaggtga tgaagcagtt gaagagaaga
     6241 agatacactg gttggggaag gctctcaaga aagctcatta acggaatcag ggataagcag
     6301 tctggaaaga caatccttga tttcctcaag tctgatggat tcgctaacag aaacttcatg
     6361 cagctcatcc acgatgattc tctcaccttt aaagaggata tccagaaggc tcaggtttca
     6421 ggacagggtg atagtctcca tgagcatatc gctaacctcg ctggatctcc tgcaatcaag
     6481 aagggaatcc tccagactgt gaaggttgtg gatgagttgg tgaaggtgat gggaaggcat
     6541 aagcctgaga acatcgtgat cgaaatggct agagagaacc agaccactca gaagggacag
     6601 aagaactcta gggaaaggat gaagaggatc gaggaaggta tcaaagagct tggatctcag
     6661 atcctcaaag agcaccctgt tgagaacact cagctccaga atgagaagct ctacctctac
     6721 tacctccaga acggaaggga tatgtatgtg gatcaagagt tggatatcaa caggctctct
     6781 gattacgatg ttgatcatat cgtgccacag tcattcttga aggatgattc tatcgataac
     6841 aaggtgctca ccaggtctga taagaacagg ggtaagagtg ataacgtgcc aagtgaagag
     6901 gttgtgaaga aaatgaagaa ctattggagg cagctcctca acgctaagct catcactcag
     6961 agaaagttcg ataacttgac taaggctgag aggggaggac tctctgaatt ggataaggca
     7021 ggattcatca agaggcagct tgtggaaacc aggcagatca ctaagcacgt tgcacagatc
     7081 ctcgattcta ggatgaacac caagtacgat gagaacgata agttgatcag ggaagtgaag
     7141 gttatcaccc tcaagtcaaa gctcgtgtct gatttcagaa aggatttcca attctacaag
     7201 gtgagggaaa tcaacaacta ccaccacgct cacgatgctt accttaacgc tgttgttgga
     7261 accgctctca tcaagaagta tcctaagctc gagtcagagt tcgtgtacgg tgattacaag
     7321 gtgtacgatg tgaggaagat gatcgctaag tctgagcaag agatcggaaa ggctaccgct
     7381 aagtatttct tctactctaa catcatgaat ttcttcaaga ccgagattac cctcgctaac
     7441 ggtgagatca gaaagaggcc actcatcgag acaaacggtg aaacaggtga gatcgtgtgg
     7501 gataagggaa gggatttcgc taccgttaga aaggtgctct ctatgccaca ggtgaacatc
     7561 gttaagaaaa ccgaggtgca gaccggtgga ttctctaaag agtctatcct ccctaagagg
     7621 aactctgata agctcattgc taggaagaag gattgggacc ctaagaaata cggtggtttc
     7681 gattctccta ccgtggctta ctctgttctc gttgtggcta aggttgagaa gggaaagagt
     7741 aagaagctca agtctgttaa ggaacttctc ggaatcacta tcatggaaag gtcatctttc
     7801 gagaagaacc caatcgattt cctcgaggct aagggataca aagaggttaa gaaggatctc
     7861 atcatcaagc tcccaaagta ctcactcttc gaactcgaga acggtagaaa gaggatgctc
     7921 gcttctgctg gtgagcttca aaagggaaac gagcttgctc tcccatctaa gtacgttaac
     7981 tttctttacc tcgcttctca ctacgagaag ttgaagggat ctccagaaga taacgagcag
     8041 aagcaacttt tcgttgagca gcacaagcac tacttggatg agatcatcga gcagatctct
     8101 gagttctcta aaagggtgat cctcgctgat gcaaacctcg ataaggtgtt gtctgcttac
     8161 aacaagcaca gagataagcc tatcagggaa caggcagaga acatcatcca tctcttcacc
     8221 cttaccaacc tcggtgctcc tgctgctttc aagtacttcg atacaaccat cgataggaag
     8281 agatacacct ctaccaaaga agtgctcgat gctaccctca tccatcagtc tatcactgga
     8341 ctctacgaga ctaggatcga tctctcacag ctcggtggtg attcaagggc tgatcctaag
     8401 aagaagagga aggtttgact cgagatatga agatgaagat gaaatatttg gtgtgtcaaa
     8461 taaaaagctt gtgtgcttaa gtttgtgttt ttttcttggc ttgttgtgtt atgaatttgt
     8521 ggctttttct aatattaaat gaatgtaaga tcacattata atgaataaac aaatgtttct
     8581 ataatccatt gtgaatgttt tgttggatct cttctgcagc atataactac tgtatgtgct
     8641 atggtatgga ctatggaata tgattaaaga taaggagctc cggtgacgga cccatggctt
     8701 cgttgaacaa cggaaactcg acttgccttc cgcacaatac atcatttctt cttagctttt
     8761 tttcttcttc ttcgttcata cagttttttt ttgtttatca gcttacattt tcttgaaccg
     8821 tagctttcgt tttcttcttt ttaactttcc attcggagtt tttgtatctt gtttcatagt
     8881 ttgtcccagg attagaatga ttaggcatcg aaccttcaag aatttgattg aataaaacat
     8941 cttcattctt aagatatgaa gataatcttc aaaaggcccc tgggaatctg aaagaagaga
     9001 agcaggccca tttatatggg aaagaacaat agtatttctt atataggccc atttaagttg
     9061 aaaacaatct tcaaaagtcc cacatcgctt agataagaaa acgaagctga gtttatatac
     9121 agctagagtc gaagtagtga ttagaggcag gtgccgctac agggcgcgtc ccattcgcca
     9181 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     9241 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     9301 tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat acgactcact atagggcgaa
     9361 ttgggtaccg ggccccccct cgaggtcctc cagcttttgt tccctttagt gagggttaat
     9421 tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     9481 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     9541 gagctaactc acattaattg cgttgcgctc actgcccgct ttccacctgc ggtcgtttta
     9601 gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc
     9661 gagtcggtgc tttttttccc ggtggcactg gccgtcgttt tacaacgtcg tgactgggaa
     9721 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     9781 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     9841 tgctagagca gcttgagctt ggatcagatt gtcgtttccc gccttcagtt taaactatca
     9901 gtgtttgaca ggatatattg gcgggtaaac ctaagagaaa agagcgttta ttagaataat
     9961 cggatattta aaagggcgtg aaaaggttta tccgttcgtc catttgtatg tgcatgccaa
    10021 ccacagggtt cccctcggga tcaaagtact ttgatccaac ccctccgctg ctatagtgca
    10081 gtcggcttct gacgttcagt gcagccgtct tctgaaaacg acatgtcgca caagtcctaa
    10141 gttacgcgac aggctgccgc cctgcccttt tcctggcgtt ttcttgtcgc gtgttttagt
    10201 cgcataaagt agaatacttg cgactagaac cggagacatt acgccatgaa caagagcgcc
    10261 gccgctggcc tgctgggcta tgcccgcgtc agcaccgacg accaggactt gaccaaccaa
    10321 cgggccgaac tgcacgcggc cggctgcacc aagctgtttt ccgagaagat caccggcacc
    10381 aggcgcgacc gcccggagct ggccaggatg cttgaccacc tacgccctgg cgacgttgtg
    10441 acagtgacca ggctagaccg cctggcccgc agcacccgcg acctactgga cattgccgag
    10501 cgcatccagg aggccggcgc gggcctgcgt agcctggcag agccgtgggc cgacaccacc
    10561 acgccggccg gccgcatggt gttgaccgtg ttcgccggca ttgccgagtt cgagcgttcc
    10621 ctaatcatcg accgcacccg gagcgggcgc gaggccgcca aggcccgagg cgtgaagttt
    10681 ggcccccgcc ctaccctcac cccggcacag atcgcgcacg cccgcgagct gatcgaccag
    10741 gaaggccgca ccgtgaaaga ggcggctgca ctgcttggcg tgcatcgctc gaccctgtac
    10801 cgcgcacttg agcgcagcga ggaagtgacg cccaccgagg ccaggcggcg cggtgccttc
    10861 cgtgaggacg cattgaccga ggccgacgcc ctggcggccg ccgagaatga acgccaagag
    10921 gaacaagcat gaaaccgcac caggacggcc aggacgaacc gtttttcatt accgaagaga
    10981 tcgaggcgga gatgatcgcg gccgggtacg tgttcgagcc gcccgcgcac gtctcaaccg
    11041 tgcggctgca tgaaatcctg gccggtttgt ctgatgccaa gctggcggcc tggccggcca
    11101 gcttggccgc tgaagaaacc gagcgccgcc gtctaaaaag gtgatgtgta tttgagtaaa
    11161 acagcttgcg tcatgcggtc gctgcgtata tgatgcgatg agtaaataaa caaatacgca
    11221 aggggaacgc atgaaggtta tcgctgtact taaccagaaa ggcgggtcag gcaagacgac
    11281 catcgcaacc catctagccc gcgccctgca actcgccggg gccgatgttc tgttagtcga
    11341 ttccgatccc cagggcagtg cccgcgattg ggcggccgtg cgggaagatc aaccgctaac
    11401 cgttgtcggc atcgaccgcc cgacgattga ccgcgacgtg aaggccatcg gccggcgcga
    11461 cttcgtagtg atcgacggag cgccccaggc ggcggacttg gctgtgtccg cgatcaaggc
    11521 agccgacttc gtgctgattc cggtgcagcc aagcccttac gacatatggg ccaccgccga
    11581 cctggtggag ctggttaagc agcgcattga ggtcacggat ggaaggctac aagcggcctt
    11641 tgtcgtgtcg cgggcgatca aaggcacgcg catcggcggt gaggttgccg aggcgctggc
    11701 cgggtacgag ctgcccattc ttgagtcccg tatcacgcag cgcgtgagct acccaggcac
    11761 tgccgccgcc ggcacaaccg ttcttgaatc agaacccgag ggcgacgctg cccgcgaggt
    11821 ccaggcgctg gccgctgaaa ttaaatcaaa actcatttga gttaatgagg taaagagaaa
    11881 atgagcaaaa gcacaaacac gctaagtgcc ggccgtccga gcgcacgcag cagcaaggct
    11941 gcaacgttgg ccagcctggc agacacgcca gccatgaagc gggtcaactt tcagttgccg
    12001 gcggaggatc acaccaagct gaagatgtac gcggtacgcc aaggcaagac cattaccgag
    12061 ctgctatctg aatacatcgc gcagctacca gagtaaatga gcaaatgaat aaatgagtag
    12121 atgaatttta gcggctaaag gaggcggcat ggaaaatcaa gaacaaccag gcaccgacgc
    12181 cgtggaatgc cccatgtgtg gaggaacggg cggttggcca ggcgtaagcg gctgggttgc
    12241 ctgccggccc tgcaatggca ctggaacccc caagcccgag gaatcggcgt gagcggtcgc
    12301 aaaccatccg gcccggtaca aatcggcgcg gcgctgggtg atgacctggt ggagaagttg
    12361 aaggccgcgc aggccgccca gcggcaacgc atcgaggcag aagcacgccc cggtgaatcg
    12421 tggcaagcgg ccgctgatcg aatccgcaaa gaatcccggc aaccgccggc agccggtgcg
    12481 ccgtcgatta ggaagccgcc caagggcgac gagcaaccag attttttcgt tccgatgctc
    12541 tatgacgtgg gcacccgcga tagtcgcagc atcatggacg tggccgtttt ccgtctgtcg
    12601 aagcgtgacc gacgagctgg cgaggtgatc cgctacgagc ttccagacgg gcacgtagag
    12661 gtttccgcag ggccggccgg catggccagt gtgtgggatt acgacctggt actgatggcg
    12721 gtttcccatc taaccgaatc catgaaccga taccgggaag ggaagggaga caagcccggc
    12781 cgcgtgttcc gtccacacgt tgcggacgta ctcaagttct gccggcgagc cgatggcgga
    12841 aagcagaaag acgacctggt agaaacctgc attcggttaa acaccacgca cgttgccatg
    12901 cagcgtacga agaaggccaa gaacggccgc ctggtgacgg tatccgaggg tgaagccttg
    12961 attagccgct acaagatcgt aaagagcgaa accgggcggc cggagtacat cgagatcgag
    13021 ctagctgatt ggatgtaccg cgagatcaca gaaggcaaga acccggacgt gctgacggtt
    13081 caccccgatt actttttgat cgatcccggc atcggccgtt ttctctaccg cctggcacgc
    13141 cgcgccgcag gcaaggcaga agccagatgg ttgttcaaga cgatctacga acgcagtggc
    13201 agcgccggag agttcaagaa gttctgtttc accgtgcgca agctgatcgg gtcaaatgac
    13261 ctgccggagt acgatttgaa ggaggaggcg gggcaggctg gcccgatcct agtcatgcgc
    13321 taccgcaacc tgatcgaggg cgaagcatcc gccggttcct aatgtacgga gcagatgcta
    13381 gggcaaattg ccctagcagg ggaaaaaggt cgaaaaggtc tctttcctgt ggatagcacg
    13441 tacattggga acccaaagcc gtacattggg aaccggaacc cgtacattgg gaacccaaag
    13501 ccgtacattg ggaaccggtc acacatgtaa gtgactgata taaaagagaa aaaaggcgat
    13561 ttttccgcct aaaactcttt aaaacttatt aaaactctta aaacccgcct ggcctgtgca
    13621 taactgtctg gccagcgcac agcccaagag ctgcaaaaag cgcctaccct tcggtcgctg
    13681 cgctccctac gccccgccgc ttcgcgtcgg cctatcgcgg ccgctggccg ctcaaaaatg
    13741 gctggcctac ggccaggcaa tctaccaggg cgcggacaag ccgcgccgtc gccactcgac
    13801 cgccggcgcc cacatcaagg caccctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
    13861 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
    13921 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca
    13981 gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta
    14041 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
    14101 atcaggccct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
    14161 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
    14221 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
    14281 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
    14341 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
    14401 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
    14461 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
    14521 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
    14581 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
    14641 gcagccactg gtaacaggat tagcagagcg aggtat
//
