LOCUS       sequence-182252-       15206 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Direct Cloning, Type: T-DNA, Engineering Reagent: 35S:AtCas9_dead + 
            AtU6:gRNA, Plant Selection: 2x35S:hpt II.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15206)
  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 15206)
  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_21B
FEATURES             Location/Qualifiers
     source          1..15206
                     /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(1827..2852)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     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     3884..3900
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4430..4775
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             join(4809..4835,4836..4838,4839..7325,7326..7328,
                     7329..8912)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             8925..8945
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      8956..9204
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     CDS             complement(9671..9976)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGTCL"
     primer_bind     9841..9857
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9867..9886
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        9867..9885
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9933..9953)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(9933..9951)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(9972..9988)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    9996..10012
                     /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(10020..10026,10027..10044,10045..10050))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10065..10086
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_RNA        10125..10200
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(10217..10233)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    10436..10460
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             11761..12390
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             12819..13892
                     /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      13958..14152
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(14388..14410)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    14496..14636
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     14551..14570
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(14651..14668)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(14822..15206,1..204))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(14902..14921)
                     /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 ccggtcggca tctactctat ttctttgccc tcggacgagt gctggggcgt
     1861 cggtttccac tatcggcgag tacttctaca cagccatcgg tccagacggc cgcgcttctg
     1921 cgggcgattt gtgtacgccc gacagtcccg gctccggatc ggacgattgc gtcgcatcga
     1981 ccctgcgccc aagctgcatc atcgaaattg ccgtcaacca agctctgata gagttggtca
     2041 agaccaatgc ggagcatata cgcccggagt cgtggcgatc ctgcaagctc cggatgcctc
     2101 cgctcgaagt agcgcgtctg ctgctccata caagccaacc acggcctcca gaagaagatg
     2161 ttggcgacct cgtattggga atccccgaac atcgcctcgc tccagtcaat gaccgctgtt
     2221 atgcggccat tgtccgtcag gacattgttg gagccgaaat ccgcgtgcac gaggtgccgg
     2281 acttcggggc agtcctcggc ccaaagcatc agctcatcga gagcctgcgc gacggacgca
     2341 ctgacggtgt cgtccatcac agtttgccag tgatacacat ggggatcagc aatcgcgcat
     2401 atgaaatcac gccatgtagt gtattgaccg attccttgcg gtccgaatgg gccgaacccg
     2461 ctcgtctggc taagatcggc cgcagcgatc gcatccatag cctccgcgac cggttgtaga
     2521 acagcgggca gttcggtttc aggcaggtct tgcaacgtga caccctgtga acggcgggag
     2581 atgcaatagg tcaggctctc gctaaactcc ccaatgtcaa gcacttccgg aatcgggagc
     2641 gcggccgatg caaagtgccg ataaacataa cgatctttgt agaaaccatc ggcgcagcta
     2701 tttacccgca ggacatatcc acgccctcct acatcgaagc tgaaagcacg agattcttcg
     2761 ccctccgaga gctgcatcag gtcggagacg ctgtcgaact tttcgatcag aaacttctcg
     2821 acagacgtcg cggtgagttc aggctttttc atatctcatt gccccccgga 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 atgattacga attcgagctc ggtacccggg gatcggcgcg ccagatttgc cttttcaatt
     3961 tcagaaagaa tgctaaccca cagatggtta gagaggctta cgcagcaggt atcatcaaga
     4021 cgatctaccc gagcaataat ctccaggaaa tcaaatacct tcccaagaag gttaaagatg
     4081 cagtcaaaag attcaggact aactgcatca agaacacaga gaaagatata tttctcaaga
     4141 tcagaagtac tattccagta tggacgattc aaggcttgct tcacaaacca aggcaagtaa
     4201 tagagattgg agtctctaaa aaggtagttc ccactgaatc aaaggccatg gagtcaaaga
     4261 ttcaaataga ggacctaaca gaactcgccg taaagactgg cgaacagttc atacagagtc
     4321 tcttacgact caatgacaag aagaaaatct tcgtcaacat ggtggagcac gacacacttg
     4381 tctactccaa aaatatcaaa gatacagtct cagaagacca aagggcaatt gagacttttc
     4441 aacaaagggt aatatccgga aacctcctcg gattccattg cccagctatc tgtcacttta
     4501 ttgtgaagat agtggaaaag gaaggtggct cctacaaatg ccatcattgc gataaaggaa
     4561 aggccatcgt tgaagatgcc tctgccgaca gtggtcccaa agatggaccc ccacccacga
     4621 ggagcatcgt ggaaaaagaa gacgttccaa ccacgtcttc aaagcaagtg gattgatgtg
     4681 atatctccac tgacgtaagg gatgacgcac aatcccacta tccttcgcaa gacccttcct
     4741 ctatataagg aagttcattt catttggaga gaacacgggg gactcctgca ggtagatcgc
     4801 tcgtcgacat ggataagaag tactctatcg gactcgctat cggaactaac tctgtgggat
     4861 gggctgtgat caccgatgag tacaaggtgc catctaagaa gttcaaggtt ctcggaaaca
     4921 ccgataggca ctctatcaag aaaaacctta tcggtgctct cctcttcgat tctggtgaaa
     4981 ctgctgaggc taccagactc aagagaaccg ctagaagaag gtacaccaga agaaagaaca
     5041 ggatctgcta cctccaagag atcttctcta acgagatggc taaagtggat gattcattct
     5101 tccacaggct cgaagagtca ttcctcgtgg aagaagataa gaagcacgag aggcacccta
     5161 tcttcggaaa catcgttgat gaggtggcat accacgagaa gtaccctact atctaccacc
     5221 tcagaaagaa gctcgttgat tctactgata aggctgatct caggctcatc tacctcgctc
     5281 tcgctcacat gatcaagttc agaggacact tcctcatcga gggtgatctc aaccctgata
     5341 actctgatgt ggataagttg ttcatccagc tcgtgcagac ctacaaccag cttttcgaag
     5401 agaaccctat caacgcttca ggtgtggatg ctaaggctat cctctctgct aggctctcta
     5461 agtcaagaag gcttgagaac ctcattgctc agctccctgg tgagaagaag aacggacttt
     5521 tcggaaactt gatcgctctc tctctcggac tcacccctaa cttcaagtct aacttcgatc
     5581 tcgctgagga tgcaaagctc cagctctcaa aggataccta cgatgatgat ctcgataacc
     5641 tcctcgctca gatcggagat cagtacgctg atttgttcct cgctgctaag aacctctctg
     5701 atgctatcct cctcagtgat atcctcagag tgaacaccga gatcaccaag gctccactct
     5761 cagcttctat gatcaagaga tacgatgagc accaccagga tctcacactt ctcaaggctc
     5821 ttgttagaca gcagctccca gagaagtaca aagagatttt cttcgatcag tctaagaacg
     5881 gatacgctgg ttacatcgat ggtggtgcat ctcaagaaga gttctacaag ttcatcaagc
     5941 ctatcctcga gaagatggat ggaaccgagg aactcctcgt gaagctcaat agagaggatc
     6001 ttctcagaaa gcagaggacc ttcgataacg gatctatccc tcatcagatc cacctcggag
     6061 agttgcacgc tatccttaga aggcaagagg atttctaccc attcctcaag gataacaggg
     6121 aaaagattga gaagattctc accttcagaa tcccttacta cgtgggacct ctcgctagag
     6181 gaaactcaag attcgcttgg atgaccagaa agtctgagga aaccatcacc ccttggaact
     6241 tcgaagaggt ggtggataag ggtgctagtg ctcagtcttt catcgagagg atgaccaact
     6301 tcgataagaa ccttccaaac gagaaggtgc tccctaagca ctctttgctc tacgagtact
     6361 tcaccgtgta caacgagttg accaaggtta agtacgtgac cgagggaatg aggaagcctg
     6421 cttttttgtc aggtgagcaa aagaaggcta tcgttgatct cttgttcaag accaacagaa
     6481 aggtgaccgt gaagcagctc aaagaggatt acttcaagaa aatcgagtgc ttcgattcag
     6541 ttgagatttc tggtgttgag gataggttca acgcatctct cggaacctac cacgatctcc
     6601 tcaagatcat taaggataag gatttcttgg ataacgagga aaacgaggat atcttggagg
     6661 atatcgttct taccctcacc ctctttgaag atagagagat gattgaagaa aggctcaaga
     6721 cctacgctca tctcttcgat gataaggtga tgaagcagtt gaagagaaga agatacactg
     6781 gttggggaag gctctcaaga aagctcatta acggaatcag ggataagcag tctggaaaga
     6841 caatccttga tttcctcaag tctgatggat tcgctaacag aaacttcatg cagctcatcc
     6901 acgatgattc tctcaccttt aaagaggata tccagaaggc tcaggtttca ggacagggtg
     6961 atagtctcca tgagcatatc gctaacctcg ctggatctcc tgcaatcaag aagggaatcc
     7021 tccagactgt gaaggttgtg gatgagttgg tgaaggtgat gggaaggcat aagcctgaga
     7081 acatcgtgat cgaaatggct agagagaacc agaccactca gaagggacag aagaactcta
     7141 gggaaaggat gaagaggatc gaggaaggta tcaaagagct tggatctcag atcctcaaag
     7201 agcaccctgt tgagaacact cagctccaga atgagaagct ctacctctac tacctccaga
     7261 acggaaggga tatgtatgtg gatcaagagt tggatatcaa caggctctct gattacgatg
     7321 ttgatgctat cgtgccacag tcattcttga aggatgattc tatcgataac aaggtgctca
     7381 ccaggtctga taagaacagg ggtaagagtg ataacgtgcc aagtgaagag gttgtgaaga
     7441 aaatgaagaa ctattggagg cagctcctca acgctaagct catcactcag agaaagttcg
     7501 ataacttgac taaggctgag aggggaggac tctctgaatt ggataaggca ggattcatca
     7561 agaggcagct tgtggaaacc aggcagatca ctaagcacgt tgcacagatc ctcgattcta
     7621 ggatgaacac caagtacgat gagaacgata agttgatcag ggaagtgaag gttatcaccc
     7681 tcaagtcaaa gctcgtgtct gatttcagaa aggatttcca attctacaag gtgagggaaa
     7741 tcaacaacta ccaccacgct cacgatgctt accttaacgc tgttgttgga accgctctca
     7801 tcaagaagta tcctaagctc gagtcagagt tcgtgtacgg tgattacaag gtgtacgatg
     7861 tgaggaagat gatcgctaag tctgagcaag agatcggaaa ggctaccgct aagtatttct
     7921 tctactctaa catcatgaat ttcttcaaga ccgagattac cctcgctaac ggtgagatca
     7981 gaaagaggcc actcatcgag acaaacggtg aaacaggtga gatcgtgtgg gataagggaa
     8041 gggatttcgc taccgttaga aaggtgctct ctatgccaca ggtgaacatc gttaagaaaa
     8101 ccgaggtgca gaccggtgga ttctctaaag agtctatcct ccctaagagg aactctgata
     8161 agctcattgc taggaagaag gattgggacc ctaagaaata cggtggtttc gattctccta
     8221 ccgtggctta ctctgttctc gttgtggcta aggttgagaa gggaaagagt aagaagctca
     8281 agtctgttaa ggaacttctc ggaatcacta tcatggaaag gtcatctttc gagaagaacc
     8341 caatcgattt cctcgaggct aagggataca aagaggttaa gaaggatctc atcatcaagc
     8401 tcccaaagta ctcactcttc gaactcgaga acggtagaaa gaggatgctc gcttctgctg
     8461 gtgagcttca aaagggaaac gagcttgctc tcccatctaa gtacgttaac tttctttacc
     8521 tcgcttctca ctacgagaag ttgaagggat ctccagaaga taacgagcag aagcaacttt
     8581 tcgttgagca gcacaagcac tacttggatg agatcatcga gcagatctct gagttctcta
     8641 aaagggtgat cctcgctgat gcaaacctcg ataaggtgtt gtctgcttac aacaagcaca
     8701 gagataagcc tatcagggaa caggcagaga acatcatcca tctcttcacc cttaccaacc
     8761 tcggtgctcc tgctgctttc aagtacttcg atacaaccat cgataggaag agatacacct
     8821 ctaccaaaga agtgctcgat gctaccctca tccatcagtc tatcactgga ctctacgaga
     8881 ctaggatcga tctctcacag ctcggtggtg attcaagggc tgatcctaag aagaagagga
     8941 aggtttgact cgagatatga agatgaagat gaaatatttg gtgtgtcaaa taaaaagctt
     9001 gtgtgcttaa gtttgtgttt ttttcttggc ttgttgtgtt atgaatttgt ggctttttct
     9061 aatattaaat gaatgtaaga tcacattata atgaataaac aaatgtttct ataatccatt
     9121 gtgaatgttt tgttggatct cttctgcagc atataactac tgtatgtgct atggtatgga
     9181 ctatggaata tgattaaaga taaggagctc cggtgacgga cccatggctt cgttgaacaa
     9241 cggaaactcg acttgccttc cgcacaatac atcatttctt cttagctttt tttcttcttc
     9301 ttcgttcata cagttttttt ttgtttatca gcttacattt tcttgaaccg tagctttcgt
     9361 tttcttcttt ttaactttcc attcggagtt tttgtatctt gtttcatagt ttgtcccagg
     9421 attagaatga ttaggcatcg aaccttcaag aatttgattg aataaaacat cttcattctt
     9481 aagatatgaa gataatcttc aaaaggcccc tgggaatctg aaagaagaga agcaggccca
     9541 tttatatggg aaagaacaat agtatttctt atataggccc atttaagttg aaaacaatct
     9601 tcaaaagtcc cacatcgctt agataagaaa acgaagctga gtttatatac agctagagtc
     9661 gaagtagtga ttagaggcag gtgccgctac agggcgcgtc ccattcgcca ttcaggctgc
     9721 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     9781 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     9841 gtaaaacgac ggccagtgag cgcgcgtaat acgactcact atagggcgaa ttgggtaccg
     9901 ggccccccct cgaggtcctc cagcttttgt tccctttagt gagggttaat tgcgcgcttg
     9961 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
    10021 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
    10081 acattaattg cgttgcgctc actgcccgct ttccacctgc ggtcgtttta gagctagaaa
    10141 tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc
    10201 tttttttccc ggtggcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
    10261 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
    10321 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgctagagca
    10381 gcttgagctt ggatcagatt gtcgtttccc gccttcagtt taaactatca gtgtttgaca
    10441 ggatatattg gcgggtaaac ctaagagaaa agagcgttta ttagaataat cggatattta
    10501 aaagggcgtg aaaaggttta tccgttcgtc catttgtatg tgcatgccaa ccacagggtt
    10561 cccctcggga tcaaagtact ttgatccaac ccctccgctg ctatagtgca gtcggcttct
    10621 gacgttcagt gcagccgtct tctgaaaacg acatgtcgca caagtcctaa gttacgcgac
    10681 aggctgccgc cctgcccttt tcctggcgtt ttcttgtcgc gtgttttagt cgcataaagt
    10741 agaatacttg cgactagaac cggagacatt acgccatgaa caagagcgcc gccgctggcc
    10801 tgctgggcta tgcccgcgtc agcaccgacg accaggactt gaccaaccaa cgggccgaac
    10861 tgcacgcggc cggctgcacc aagctgtttt ccgagaagat caccggcacc aggcgcgacc
    10921 gcccggagct ggccaggatg cttgaccacc tacgccctgg cgacgttgtg acagtgacca
    10981 ggctagaccg cctggcccgc agcacccgcg acctactgga cattgccgag cgcatccagg
    11041 aggccggcgc gggcctgcgt agcctggcag agccgtgggc cgacaccacc acgccggccg
    11101 gccgcatggt gttgaccgtg ttcgccggca ttgccgagtt cgagcgttcc ctaatcatcg
    11161 accgcacccg gagcgggcgc gaggccgcca aggcccgagg cgtgaagttt ggcccccgcc
    11221 ctaccctcac cccggcacag atcgcgcacg cccgcgagct gatcgaccag gaaggccgca
    11281 ccgtgaaaga ggcggctgca ctgcttggcg tgcatcgctc gaccctgtac cgcgcacttg
    11341 agcgcagcga ggaagtgacg cccaccgagg ccaggcggcg cggtgccttc cgtgaggacg
    11401 cattgaccga ggccgacgcc ctggcggccg ccgagaatga acgccaagag gaacaagcat
    11461 gaaaccgcac caggacggcc aggacgaacc gtttttcatt accgaagaga tcgaggcgga
    11521 gatgatcgcg gccgggtacg tgttcgagcc gcccgcgcac gtctcaaccg tgcggctgca
    11581 tgaaatcctg gccggtttgt ctgatgccaa gctggcggcc tggccggcca gcttggccgc
    11641 tgaagaaacc gagcgccgcc gtctaaaaag gtgatgtgta tttgagtaaa acagcttgcg
    11701 tcatgcggtc gctgcgtata tgatgcgatg agtaaataaa caaatacgca aggggaacgc
    11761 atgaaggtta tcgctgtact taaccagaaa ggcgggtcag gcaagacgac catcgcaacc
    11821 catctagccc gcgccctgca actcgccggg gccgatgttc tgttagtcga ttccgatccc
    11881 cagggcagtg cccgcgattg ggcggccgtg cgggaagatc aaccgctaac cgttgtcggc
    11941 atcgaccgcc cgacgattga ccgcgacgtg aaggccatcg gccggcgcga cttcgtagtg
    12001 atcgacggag cgccccaggc ggcggacttg gctgtgtccg cgatcaaggc agccgacttc
    12061 gtgctgattc cggtgcagcc aagcccttac gacatatggg ccaccgccga cctggtggag
    12121 ctggttaagc agcgcattga ggtcacggat ggaaggctac aagcggcctt tgtcgtgtcg
    12181 cgggcgatca aaggcacgcg catcggcggt gaggttgccg aggcgctggc cgggtacgag
    12241 ctgcccattc ttgagtcccg tatcacgcag cgcgtgagct acccaggcac tgccgccgcc
    12301 ggcacaaccg ttcttgaatc agaacccgag ggcgacgctg cccgcgaggt ccaggcgctg
    12361 gccgctgaaa ttaaatcaaa actcatttga gttaatgagg taaagagaaa atgagcaaaa
    12421 gcacaaacac gctaagtgcc ggccgtccga gcgcacgcag cagcaaggct gcaacgttgg
    12481 ccagcctggc agacacgcca gccatgaagc gggtcaactt tcagttgccg gcggaggatc
    12541 acaccaagct gaagatgtac gcggtacgcc aaggcaagac cattaccgag ctgctatctg
    12601 aatacatcgc gcagctacca gagtaaatga gcaaatgaat aaatgagtag atgaatttta
    12661 gcggctaaag gaggcggcat ggaaaatcaa gaacaaccag gcaccgacgc cgtggaatgc
    12721 cccatgtgtg gaggaacggg cggttggcca ggcgtaagcg gctgggttgc ctgccggccc
    12781 tgcaatggca ctggaacccc caagcccgag gaatcggcgt gagcggtcgc aaaccatccg
    12841 gcccggtaca aatcggcgcg gcgctgggtg atgacctggt ggagaagttg aaggccgcgc
    12901 aggccgccca gcggcaacgc atcgaggcag aagcacgccc cggtgaatcg tggcaagcgg
    12961 ccgctgatcg aatccgcaaa gaatcccggc aaccgccggc agccggtgcg ccgtcgatta
    13021 ggaagccgcc caagggcgac gagcaaccag attttttcgt tccgatgctc tatgacgtgg
    13081 gcacccgcga tagtcgcagc atcatggacg tggccgtttt ccgtctgtcg aagcgtgacc
    13141 gacgagctgg cgaggtgatc cgctacgagc ttccagacgg gcacgtagag gtttccgcag
    13201 ggccggccgg catggccagt gtgtgggatt acgacctggt actgatggcg gtttcccatc
    13261 taaccgaatc catgaaccga taccgggaag ggaagggaga caagcccggc cgcgtgttcc
    13321 gtccacacgt tgcggacgta ctcaagttct gccggcgagc cgatggcgga aagcagaaag
    13381 acgacctggt agaaacctgc attcggttaa acaccacgca cgttgccatg cagcgtacga
    13441 agaaggccaa gaacggccgc ctggtgacgg tatccgaggg tgaagccttg attagccgct
    13501 acaagatcgt aaagagcgaa accgggcggc cggagtacat cgagatcgag ctagctgatt
    13561 ggatgtaccg cgagatcaca gaaggcaaga acccggacgt gctgacggtt caccccgatt
    13621 actttttgat cgatcccggc atcggccgtt ttctctaccg cctggcacgc cgcgccgcag
    13681 gcaaggcaga agccagatgg ttgttcaaga cgatctacga acgcagtggc agcgccggag
    13741 agttcaagaa gttctgtttc accgtgcgca agctgatcgg gtcaaatgac ctgccggagt
    13801 acgatttgaa ggaggaggcg gggcaggctg gcccgatcct agtcatgcgc taccgcaacc
    13861 tgatcgaggg cgaagcatcc gccggttcct aatgtacgga gcagatgcta gggcaaattg
    13921 ccctagcagg ggaaaaaggt cgaaaaggtc tctttcctgt ggatagcacg tacattggga
    13981 acccaaagcc gtacattggg aaccggaacc cgtacattgg gaacccaaag ccgtacattg
    14041 ggaaccggtc acacatgtaa gtgactgata taaaagagaa aaaaggcgat ttttccgcct
    14101 aaaactcttt aaaacttatt aaaactctta aaacccgcct ggcctgtgca taactgtctg
    14161 gccagcgcac agccgaagag ctgcaaaaag cgcctaccct tcggtcgctg cgctccctac
    14221 gccccgccgc ttcgcgtcgg cctatcgcgg ccgctggccg ctcaaaaatg gctggcctac
    14281 ggccaggcaa tctaccaggg cgcggacaag ccgcgccgtc gccactcgac cgccggcgcc
    14341 cacatcaagg caccctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
    14401 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
    14461 cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc
    14521 gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
    14581 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct
    14641 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
    14701 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
    14761 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
    14821 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
    14881 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
    14941 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
    15001 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
    15061 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
    15121 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
    15181 gtaacaggat tagcagagcg aggtat
//
