LOCUS       sequence-208235-       14954 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCLV3:Cas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14954)
  TITLE     pJKW0457
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 14954)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pJKW0474
FEATURES             Location/Qualifiers
     source          1..14954
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..25
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(111..363)
                     /label=MAS terminator
                     /note="mannopine synthase terminator"
     CDS             complement(373..924)
                     /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(930..1310)
                     /label=MAS promoter
                     /note="mannopine synthase promoter (Velten et al., 1984)"
     CDS             2806..2871
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2878..2898
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2923..7023
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             2923..7023
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7024..7071
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(8456..8473)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8456..8472)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8465..8487)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    8675..8699
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             10000..10629
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             11058..12131
                     /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      12197..12391
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(12627..12649)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    12735..12875
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     12790..12809
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(12890..12907)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13061..13649)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13141..13160)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(13736..14530)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
ORIGIN
        1 tggcaggata tattgtggtg taaacaaatt gacgcttaga caacttaata acacattgcg
       61 gacgttttta atgtactgaa ttaacgccga attaattcga gctcggatct gataatttat
      121 ttgaaaattc ataagaaaag caaacgttac atgaattgat gaaacaatac aaagacagat
      181 aaagccacgc acatttagga tattggccga gattactgaa tattgagtaa gatcacggaa
      241 tttctgacag gagcatgtct tcaattcagc ccaaatggca gttgaaatac tcaaaccgcc
      301 ccatatgcag gagcggatca ttcattgttt gtttggttgc ctttgccaac atgggagtcc
      361 aagattctgc agtcaaatct cggtgacggg caggaccgga cggggcggta ccggcaggct
      421 gaagtccagc tgccagaaac ccacgtcatg ccagttcccg tgcttgaagc cggccgcccg
      481 cagcatgccg cggggggcat atccgagcgc ctcgtgcatg cgcacgctcg ggtcgttggg
      541 cagcccgatg acagcgacca cgctcttgaa gccctgtgcc tccagggact tcagcaggtg
      601 ggtgtagagc gtggagccca gtcccgtccg ctggtggcgg ggggagacgt acacggtcga
      661 ctcggccgtc cagtcgtagg cgttgcgtgc cttccagggg cccgcgtagg cgatgccggc
      721 gacctcgccg tccacctcgg cgacgagcca gggatagcgc tcccgcagac ggacgaggtc
      781 gtccgtccac tcctgcggtt cctgcggctc ggtacggaag ttgaccgtgc ttgtctcgat
      841 gtagtggttg acgatggtgc agaccgccgg catgtccgcc tcggtggcac ggcggatgtc
      901 ggccgggcgt cgttctgggc tcatcgattc gatttggtgt atcgagattg gttatgaaat
      961 tcagatgcta gtgtaatgta ttggtaattt gggaagatat aataggaagc aaggctattt
     1021 atccatttct gaaaaggcga aatggcgtca ccgcgagcgt cacgcgcatt ccgttcttgc
     1081 tgtaaagcgt tgtttggtac acttttgact agcgaggctt ggcgtgtcag cgtatctatt
     1141 caaaagtcgt taatggctgc ggatcaagaa aaagttggaa tagaaacaga atacccgcga
     1201 aattcaggcc cggttgccat gtcctacacg ccgaaataaa cgaccaaatt agtagaaaaa
     1261 taaaaactga ctcggatact tacgtcacgt cttgcgcact gatttgaaaa atctcagaat
     1321 tccggattat ccataataaa aacaaaacta gatacatttt ctaagtacat taacacataa
     1381 gaatatcatt tttgtactac caaaaaaaaa gtaaaaactt atgtccatgg tattattata
     1441 attggtggta aatcagtaaa ttagttgtta gcaaaaaaat aaaatttagt taaaaagtag
     1501 tggcacctta ttggcccaaa agagtaacga agcaaaacgg aatctgaaaa ctgttaaaac
     1561 ttaaaagaat agataattga gatataatat cgtatgatcg gacggctgtg atctgatgcc
     1621 atcggatggg cacgtgtcag aagtgtactc caggtatcat tctctctgcc tcatgacatc
     1681 agccgatggt accctcgata cggcgccggc aaggctcata taatccattc aatttatgtt
     1741 ttttctgcta gcaaatagat ttgccattga tttgatactc tcaataatat tatccgatta
     1801 aggtatcatc catattgttc aatataattt aagcatataa ctgtttccag attaaacaat
     1861 ataatttata agagcaactg taatacttta ctttaaaagt tttcaaatca gaatctcttt
     1921 tctttttcta caaatctgga aacaatttaa tccatattgt ttttgttttt tttaccttct
     1981 catatttaga tgctatttat tattgtgaca caagcttagt ttaatttctt gttggttaca
     2041 tttttttttc taaatacaca aatattatat gtttaatatt atttcacaca tttcttagaa
     2101 gaaatctaaa tacatgaaat tatatttatt aaaaagaagt ctaaactata tatgtattta
     2161 aatggaattt taaatatggg tagtaatata gaaacaccat tgatatatta gagtatgtgc
     2221 cggtgccgta tacatactgt tgtattatag taacctatat atggtatacc tagtcaatgt
     2281 gtacggtatt tttcatatgt attagttgtg aacctccaca gcatgttaga cttaggaatt
     2341 aattataact gaaccagaac tttgagtcta attatacccc tgccgtttta tacaactttg
     2401 aatattgatt ccagtggcta atttaaataa gttaaaggaa acaaaaactg caagtaagtg
     2461 aaaaatacac aattgtaagt aaatgttaga tatttaattt atttatagct aaatcatgaa
     2521 caagttcgta taagatctag atatatgtac catatacctt ttctatgccc actatatata
     2581 cttactacac ataatatata aaccaaccaa aatatttgaa tagttaataa ctatgataca
     2641 cgtttaggac aaataaaatt aaaaaatagg agatcccatt tctcgccctt gtaggcttac
     2701 gctataaatt ggactgtccc cttctcattt cattaccaaa gtaaagaaca gtttctatat
     2761 ttctctcttt atctctctca ctcagtcact ttctctctaa aaatggacta taaggaccac
     2821 gacggagact acaaggatca tgatattgat tacaaagacg atgacgataa gatggcccca
     2881 aagaagaagc ggaaggtcgg tatccacgga gtcccagcag ccgacaagaa gtacagcatc
     2941 ggcctggaca tcggcaccaa ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg
     3001 cccagcaaga aattcaaggt gctgggcaac accgaccggc acagcatcaa gaagaacctg
     3061 atcggagccc tgctgttcga cagcggcgaa acagccgagg ccacccggct gaagagaacc
     3121 gccagaagaa gatacaccag acggaagaac cggatctgct atctgcaaga gatcttcagc
     3181 aacgagatgg ccaaggtgga cgacagcttc ttccacagac tggaagagtc cttcctggtg
     3241 gaagaggata agaagcacga gcggcacccc atcttcggca acatcgtgga cgaggtggcc
     3301 taccacgaga agtaccccac catctaccac ctgagaaaga aactggtgga cagcaccgac
     3361 aaggccgacc tgcggctgat ctatctggcc ctggcccaca tgatcaagtt ccggggccac
     3421 ttcctgatcg agggcgacct gaaccccgac aacagcgacg tggacaagct gttcatccag
     3481 ctggtgcaga cctacaacca gctgttcgag gaaaacccca tcaacgccag cggcgtggac
     3541 gccaaggcca tcctgtctgc cagactgagc aagagcagac ggctggaaaa tctgatcgcc
     3601 cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc tgattgccct gagcctgggc
     3661 ctgaccccca acttcaagag caacttcgac ctggccgagg atgccaaact gcagctgagc
     3721 aaggacacct acgacgacga cctggacaac ctgctggccc agatcggcga ccagtacgcc
     3781 gacctgtttc tggccgccaa gaacctgtcc gacgccatcc tgctgagcga catcctgaga
     3841 gtgaacaccg agatcaccaa ggcccccctg agcgcctcta tgatcaagag atacgacgag
     3901 caccaccagg acctgaccct gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac
     3961 aaagagattt tcttcgacca gagcaagaac ggctacgccg gctacattga cggcggagcc
     4021 agccaggaag agttctacaa gttcatcaag cccatcctgg aaaagatgga cggcaccgag
     4081 gaactgctcg tgaagctgaa cagagaggac ctgctgcgga agcagcggac cttcgacaac
     4141 ggcagcatcc cccaccagat ccacctggga gagctgcacg ccattctgcg gcggcaggaa
     4201 gatttttacc cattcctgaa ggacaaccgg gaaaagatcg agaagatcct gaccttccgc
     4261 atcccctact acgtgggccc tctggccagg ggaaacagca gattcgcctg gatgaccaga
     4321 aagagcgagg aaaccatcac cccctggaac ttcgaggaag tggtggacaa gggcgcttcc
     4381 gcccagagct tcatcgagcg gatgaccaac ttcgataaga acctgcccaa cgagaaggtg
     4441 ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt ataacgagct gaccaaagtg
     4501 aaatacgtga ccgagggaat gagaaagccc gccttcctga gcggcgagca gaaaaaggcc
     4561 atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac
     4621 tacttcaaga aaatcgagtg cttcgactcc gtggaaatct ccggcgtgga agatcggttc
     4681 aacgcctccc tgggcacata ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg
     4741 gacaatgagg aaaacgagga cattctggaa gatatcgtgc tgaccctgac actgtttgag
     4801 gacagagaga tgatcgagga acggctgaaa acctatgccc acctgttcga cgacaaagtg
     4861 atgaagcagc tgaagcggcg gagatacacc ggctggggca ggctgagccg gaagctgatc
     4921 aacggcatcc gggacaagca gtccggcaag acaatcctgg atttcctgaa gtccgacggc
     4981 ttcgccaaca gaaacttcat gcagctgatc cacgacgaca gcctgacctt taaagaggac
     5041 atccagaaag cccaggtgtc cggccagggc gatagcctgc acgagcacat tgccaatctg
     5101 gccggcagcc ccgccattaa gaagggcatc ctgcagacag tgaaggtggt ggacgagctc
     5161 gtgaaagtga tgggccggca caagcccgag aacatcgtga tcgaaatggc cagagagaac
     5221 cagaccaccc agaagggaca gaagaacagc cgcgagagaa tgaagcggat cgaagagggc
     5281 atcaaagagc tgggcagcca gatcctgaaa gaacaccccg tggaaaacac ccagctgcag
     5341 aacgagaagc tgtacctgta ctacctgcag aatgggcggg atatgtacgt ggaccaggaa
     5401 ctggacatca accggctgtc cgactacgat gtggaccata tcgtgcctca gagctttctg
     5461 aaggacgact ccatcgacaa caaggtgctg accagaagcg acaagaaccg gggcaagagc
     5521 gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga actactggcg gcagctgctg
     5581 aacgccaagc tgattaccca gagaaagttc gacaatctga ccaaggccga gagaggcggc
     5641 ctgagcgaac tggataaggc cggcttcatc aagagacagc tggtggaaac ccggcagatc
     5701 acaaagcacg tggcacagat cctggactcc cggatgaaca ctaagtacga cgagaatgac
     5761 aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca agctggtgtc cgatttccgg
     5821 aaggatttcc agttttacaa agtgcgcgag atcaacaact accaccacgc ccacgacgcc
     5881 tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt accctaagct ggaaagcgag
     5941 ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag
     6001 gaaatcggca aggctaccgc caagtacttc ttctacagca acatcatgaa ctttttcaag
     6061 accgagatta ccctggccaa cggcgagatc cggaagcggc ctctgatcga gacaaacggc
     6121 gaaaccgggg agatcgtgtg ggataagggc cgggattttg ccaccgtgcg gaaagtgctg
     6181 agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa
     6241 gagtctatcc tgcccaagag gaacagcgat aagctgatcg ccagaaagaa ggactgggac
     6301 cctaagaagt acggcggctt cgacagcccc accgtggcct attctgtgct ggtggtggcc
     6361 aaagtggaaa agggcaagtc caagaaactg aagagtgtga aagagctgct ggggatcacc
     6421 atcatggaaa gaagcagctt cgagaagaat cccatcgact ttctggaagc caagggctac
     6481 aaagaagtga aaaaggacct gatcatcaag ctgcctaagt actccctgtt cgagctggaa
     6541 aacggccgga agagaatgct ggcctctgcc ggcgaactgc agaagggaaa cgaactggcc
     6601 ctgccctcca aatatgtgaa cttcctgtac ctggccagcc actatgagaa gctgaagggc
     6661 tcccccgagg ataatgagca gaaacagctg tttgtggaac agcacaagca ctacctggac
     6721 gagatcatcg agcagatcag cgagttctcc aagagagtga tcctggccga cgctaatctg
     6781 gacaaagtgc tgtccgccta caacaagcac cgggataagc ccatcagaga gcaggccgag
     6841 aatatcatcc acctgtttac cctgaccaat ctgggagccc ctgccgcctt caagtacttt
     6901 gacaccacca tcgaccggaa gaggtacacc agcaccaaag aggtgctgga cgccaccctg
     6961 atccaccaga gcatcaccgg cctgtacgag acacggatcg acctgtctca gctgggaggc
     7021 gacaaaaggc cggcggccac gaaaaaggcc ggccaggcaa aaaagaaaaa gtaacctaat
     7081 ctcttgttgc tttaaattat ttcatattgt aaattacttt ctgctttatc ggttttacca
     7141 tttcgggagt cttttttgtg tgcaatctgt ttcgtttggt aagcttgtag tttcatgaaa
     7201 gtgaatgtaa gatatgcatt acgtttgttg ctgaagtgaa tgtaagatac gcactattat
     7261 atctcatgat tttctaagaa aaccctctta aaacgaagat gtctatagca ttacgtttct
     7321 atttccatat aatacgttaa aatttatggt ttttacgtat aaaatgcaaa ataaagacac
     7381 aagtatatct ccaaagcaat gtaccgttgg gaaaatttat tagtacgttt tcaattgtca
     7441 atgcaaataa ttaatggatg tgatagtcac aattaaacat acaataataa aaatgatgat
     7501 gatgattcga tgatgtggtg ggaaggataa attaaaccga ctttggggca gtgacaggca
     7561 gtgtcagtgt caaagacaac catttgtagt cactatttct atcgaaggtt gcaaattgaa
     7621 tggtggagga gtatcaaaac gacacacata cttgaaaaga tattttaata atataaaaaa
     7681 attggtgatg gcgtaataac aaacctagag ctaattatta tccttaatga taccaaatct
     7741 atatgatacg atatttgttt taaaaagagt aaagactgac acttgagatg tgacactggc
     7801 gatttcgctc acgtcaccac ttttcccacc tcaaataacg cttacggctt tatccattaa
     7861 ttctaagtat aattttaagt gtattttttc ttgccaaatt caaatatatc ttactaaatg
     7921 gatgaacatt ataaaattgt tatcaaaacc attaaatgtt cttataattt ctttcgttcc
     7981 tccaatgtca tcccaagact ttttgaccta atatatgata tatctaactt gctttggaat
     8041 cgtatgacat atatcttcaa atacatattt cgtatttttt tttcacgaaa actaatttag
     8101 aaagtagaaa accagctatt ttaaagaaaa taaagtgtgt ttatatatat tctaaaacaa
     8161 tgctataaga acataagacc aagatatata caatgttatt ttatatttat tattaagcat
     8221 taacattgaa attaaaaata ttaaacatgt ataccaaagt aatcaacatt gtagttatta
     8281 ctactctctc tgttcatttt tgtttgattg tttagaaaaa acacacatat taagaaaaca
     8341 tattaaatat tgattataaa tgtattattt ttaatgtttt acagttttct ataactttaa
     8401 accaatgata attaactatt tttttaaaaa attcctgcag gcatgcaagc ttggcactgg
     8461 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     8521 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     8581 cccaacagtt gcgcagcctg aatggcgaat gctagagcag cttgagcttg gatcagattg
     8641 tcgtttcccg ccttcagttt aaactatcag tgtttgacag gatatattgg cgggtaaacc
     8701 taagagaaaa gagcgtttat tagaataatc ggatatttaa aagggcgtga aaaggtttat
     8761 ccgttcgtcc atttgtatgt gcatgccaac cacagggttc ccctcgggat caaagtactt
     8821 tgatccaacc cctccgctgc tatagtgcag tcggcttctg acgttcagtg cagccgtctt
     8881 ctgaaaacga catgtcgcac aagtcctaag ttacgcgaca ggctgccgcc ctgccctttt
     8941 cctggcgttt tcttgtcgcg tgttttagtc gcataaagta gaatacttgc gactagaacc
     9001 ggagacatta cgccatgaac aagagcgccg ccgctggcct gctgggctat gcccgcgtca
     9061 gcaccgacga ccaggacttg accaaccaac gggccgaact gcacgcggcc ggctgcacca
     9121 agctgttttc cgagaagatc accggcacca ggcgcgaccg cccggagctg gccaggatgc
     9181 ttgaccacct acgccctggc gacgttgtga cagtgaccag gctagaccgc ctggcccgca
     9241 gcacccgcga cctactggac attgccgagc gcatccagga ggccggcgcg ggcctgcgta
     9301 gcctggcaga gccgtgggcc gacaccacca cgccggccgg ccgcatggtg ttgaccgtgt
     9361 tcgccggcat tgccgagttc gagcgttccc taatcatcga ccgcacccgg agcgggcgcg
     9421 aggccgccaa ggcccgaggc gtgaagtttg gcccccgccc taccctcacc ccggcacaga
     9481 tcgcgcacgc ccgcgagctg atcgaccagg aaggccgcac cgtgaaagag gcggctgcac
     9541 tgcttggcgt gcatcgctcg accctgtacc gcgcacttga gcgcagcgag gaagtgacgc
     9601 ccaccgaggc caggcggcgc ggtgccttcc gtgaggacgc attgaccgag gccgacgccc
     9661 tggcggccgc cgagaatgaa cgccaagagg aacaagcatg aaaccgcacc aggacggcca
     9721 ggacgaaccg tttttcatta ccgaagagat cgaggcggag atgatcgcgg ccgggtacgt
     9781 gttcgagccg cccgcgcacg tctcaaccgt gcggctgcat gaaatcctgg ccggtttgtc
     9841 tgatgccaag ctggcggcct ggccggccag cttggccgct gaagaaaccg agcgccgccg
     9901 tctaaaaagg tgatgtgtat ttgagtaaaa cagcttgcgt catgcggtcg ctgcgtatat
     9961 gatgcgatga gtaaataaac aaatacgcaa ggggaacgca tgaaggttat cgctgtactt
    10021 aaccagaaag gcgggtcagg caagacgacc atcgcaaccc atctagcccg cgccctgcaa
    10081 ctcgccgggg ccgatgttct gttagtcgat tccgatcccc agggcagtgc ccgcgattgg
    10141 gcggccgtgc gggaagatca accgctaacc gttgtcggca tcgaccgccc gacgattgac
    10201 cgcgacgtga aggccatcgg ccggcgcgac ttcgtagtga tcgacggagc gccccaggcg
    10261 gcggacttgg ctgtgtccgc gatcaaggca gccgacttcg tgctgattcc ggtgcagcca
    10321 agcccttacg acatatgggc caccgccgac ctggtggagc tggttaagca gcgcattgag
    10381 gtcacggatg gaaggctaca agcggccttt gtcgtgtcgc gggcgatcaa aggcacgcgc
    10441 atcggcggtg aggttgccga ggcgctggcc gggtacgagc tgcccattct tgagtcccgt
    10501 atcacgcagc gcgtgagcta cccaggcact gccgccgccg gcacaaccgt tcttgaatca
    10561 gaacccgagg gcgacgctgc ccgcgaggtc caggcgctgg ccgctgaaat taaatcaaaa
    10621 ctcatttgag ttaatgaggt aaagagaaaa tgagcaaaag cacaaacacg ctaagtgccg
    10681 gccgtccgag cgcacgcagc agcaaggctg caacgttggc cagcctggca gacacgccag
    10741 ccatgaagcg ggtcaacttt cagttgccgg cggaggatca caccaagctg aagatgtacg
    10801 cggtacgcca aggcaagacc attaccgagc tgctatctga atacatcgcg cagctaccag
    10861 agtaaatgag caaatgaata aatgagtaga tgaattttag cggctaaagg aggcggcatg
    10921 gaaaatcaag aacaaccagg caccgacgcc gtggaatgcc ccatgtgtgg aggaacgggc
    10981 ggttggccag gcgtaagcgg ctgggttgcc tgccggccct gcaatggcac tggaaccccc
    11041 aagcccgagg aatcggcgtg agcggtcgca aaccatccgg cccggtacaa atcggcgcgg
    11101 cgctgggtga tgacctggtg gagaagttga aggccgcgca ggccgcccag cggcaacgca
    11161 tcgaggcaga agcacgcccc ggtgaatcgt ggcaagcggc cgctgatcga atccgcaaag
    11221 aatcccggca accgccggca gccggtgcgc cgtcgattag gaagccgccc aagggcgacg
    11281 agcaaccaga ttttttcgtt ccgatgctct atgacgtggg cacccgcgat agtcgcagca
    11341 tcatggacgt ggccgttttc cgtctgtcga agcgtgaccg acgagctggc gaggtgatcc
    11401 gctacgagct tccagacggg cacgtagagg tttccgcagg gccggccggc atggccagtg
    11461 tgtgggatta cgacctggta ctgatggcgg tttcccatct aaccgaatcc atgaaccgat
    11521 accgggaagg gaagggagac aagcccggcc gcgtgttccg tccacacgtt gcggacgtac
    11581 tcaagttctg ccggcgagcc gatggcggaa agcagaaaga cgacctggta gaaacctgca
    11641 ttcggttaaa caccacgcac gttgccatgc agcgtacgaa gaaggccaag aacggccgcc
    11701 tggtgacggt atccgagggt gaagccttga ttagccgcta caagatcgta aagagcgaaa
    11761 ccgggcggcc ggagtacatc gagatcgagc tagctgattg gatgtaccgc gagatcacag
    11821 aaggcaagaa cccggacgtg ctgacggttc accccgatta ctttttgatc gatcccggca
    11881 tcggccgttt tctctaccgc ctggcacgcc gcgccgcagg caaggcagaa gccagatggt
    11941 tgttcaagac gatctacgaa cgcagtggca gcgccggaga gttcaagaag ttctgtttca
    12001 ccgtgcgcaa gctgatcggg tcaaatgacc tgccggagta cgatttgaag gaggaggcgg
    12061 ggcaggctgg cccgatccta gtcatgcgct accgcaacct gatcgagggc gaagcatccg
    12121 ccggttccta atgtacggag cagatgctag ggcaaattgc cctagcaggg gaaaaaggtc
    12181 gaaaaggtct ctttcctgtg gatagcacgt acattgggaa cccaaagccg tacattggga
    12241 accggaaccc gtacattggg aacccaaagc cgtacattgg gaaccggtca cacatgtaag
    12301 tgactgatat aaaagagaaa aaaggcgatt tttccgccta aaactcttta aaacttatta
    12361 aaactcttaa aacccgcctg gcctgtgcat aactgtctgg ccagcgcaca gccgaagagc
    12421 tgcaaaaagc gcctaccctt cggtcgctgc gctccctacg ccccgccgct tcgcgtcggc
    12481 ctatcgcggc cgctggccgc tcaaaaatgg ctggcctacg gccaggcaat ctaccagggc
    12541 gcggacaagc cgcgccgtcg ccactcgacc gccggcgccc acatcaaggc accctgcctc
    12601 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
    12661 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
    12721 ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc
    12781 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
    12841 cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg
    12901 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
    12961 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
    13021 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
    13081 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
    13141 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
    13201 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
    13261 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
    13321 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
    13381 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
    13441 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
    13501 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
    13561 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
    13621 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
    13681 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgcattct aggtactaaa
    13741 acaattcatc cagtaaaata taatatttta ttttctccca atcaggcttg atccccagta
    13801 agtcaaaaaa tagctcgaca tactgttctt ccccgatatc ctccctgatc gaccggacgc
    13861 agaaggcaat gtcataccac ttgtccgccc tgccgcttct cccaagatca ataaagccac
    13921 ttactttgcc atctttcaca aagatgttgc tgtctcccag gtcgccgtgg gaaaagacaa
    13981 gttcctcttc gggcttttcc gtctttaaaa aatcatacag ctcgcgcgga tctttaaatg
    14041 gagtgtcttc ttcccagttt tcgcaatcca catcggccag atcgttattc agtaagtaat
    14101 ccaattcggc taagcggctg tctaagctat tcgtataggg acaatccgat atgtcgatgg
    14161 agtgaaagag cctgatgcac tccgcataca gctcgataat cttttcaggg ctttgttcat
    14221 cttcatactc ttccgagcaa aggacgccat cggcctcact catgagcaga ttgctccagc
    14281 catcatgccg ttcaaagtgc aggacctttg gaacaggcag ctttccttcc agccatagca
    14341 tcatgtcctt ttcccgttcc acatcatagg tggtcccttt ataccggctg tccgtcattt
    14401 ttaaatatag gttttcattt tctcccacca gcttatatac cttagcagga gacattcctt
    14461 ccgtatcttt tacgcagcgg tatttttcga tcagtttttt caattccggt gatattctca
    14521 ttttagccat ttattatttc cttcctcttt tctacagtat ttaaagatac cccaagaagc
    14581 taattataac aagacgaact ccaattcact gttccttgca ttctaaaacc ttaaatacca
    14641 gaaaacagct ttttcaaagt tgttttcaaa gttggcgtat aacatagtat cgacggagcc
    14701 gattttgaaa ccgcggtgat cacaggcagc aacgctctgt catcgttaca atcaacatgc
    14761 taccctccgc gagatcatcc gtgtttcaaa cccggcagct tagttgccgt tcttccgaat
    14821 agcatcggta acatgagcaa agtctgccgc cttacaacgg ctctcccgct gacgccgtcc
    14881 cggactgatg ggctgcctgt atcgagtggt gattttgtgc cgagctgccg gtcggggagc
    14941 tgttggctgg ctgg
//
