LOCUS       sequence-373253-       16692 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Binary vector for CRISPR/Cas9 (target 2: Mpphot [negative control]) 
            in plants (for Agrobacterium-mediated genetic transformation)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16692)
  AUTHORS   Konno R, Tanaka H, Kodama Y
  TITLE     SKLPT imaging: Efficient in vivo pre-evaluation of genome-editing 
            modules using fluorescent protein with peroxisome targeting signal.
  JOURNAL   Biochem Biophys Res Commun. 2018 Sep 3;503(1):235-241. doi: 
            10.1016/j.bbrc.2018.06.008. Epub 2018 Jun 12.
  PUBMED    29885839
REFERENCE   2  (bases 1 to 16692)
  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"; doi: 
            "10.1016/j.bbrc.2018.06.008"; journalName: "Biochem Biophys Res 
            Commun"; date: "2018-09-3- 3"; volume: "503"; issue: "1"; pages: 
            "235-241"
COMMENT     Sequence Label: pMpGE010-Target2 (Mpphot)
FEATURES             Location/Qualifiers
     source          1..16692
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    54..78
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     266..288
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     280..297
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     281..297
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2057..2081
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2102..6205
                     /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             2102..6205
                     /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             6218..6238
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(6741..6765)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    6794..6818
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        7371..7446
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(7488..7512)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      7552..7804
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(7837..7853)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7837..7853)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7850..7872)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7861..7877
                     /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(7885..7891,7892..7909,7910..7915))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7930..7951
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     polyA_signal    8036..8210
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(8251..9276)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     promoter        complement(9343..10019)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     complement(9386..9406)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     misc_feature    10214..10238
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             10759..11550
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      11796..12384
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12285..12304
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12538..12555
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    12570..12710
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(12636..12655)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     12796..12818
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      13054..13248
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(13314..14387)
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     CDS             complement(14816..15445)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
ORIGIN
        1 tttcacgccc ttttaaatat ccgattattc taataaacgc tcttttctct taggtttacc
       61 cgccaatata tcctgtcaaa cactgatagt ttaaactgaa ggcgggaaac gacaatctga
      121 tccaagctca agctgctcta gcattcgcca ttcaggctgc gcaactgttg ggaagggcga
      181 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
      241 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc
      301 aagcttggcc cgggcaaatg agtcacacac attgttgaga gacatatcaa aataactcaa
      361 ttgtaaggta agttgataga tttcccagtt aaatgcatta ttttaacaat atttgtatct
      421 caaattttca aaatcatatc aaatttgcag atcttgatga tgattcacaa ctctgtcttc
      481 gatttttgat tgttaacggg tatcgatatt tgggtaactt aaaaagaatt tgctaaagat
      541 cttaaaaaac agattaaaaa aacctttcaa aatagctaat cctttaaatc cgtttcacaa
      601 tataatgctc attgatgata atcgaaaaga ttccatttta aagtgcatat aaattgctta
      661 attatctctt tctgtctcgt aatattttgt tctctcctat tggatcggac ccgcatcttc
      721 tcgatcagtt cactatacta atatttggtc aacttcaaga gagattagaa cttcgttctc
      781 caaagtgtaa aggccgacaa gcagaaaagg aaacttcaat cagaagcctt ctaggttttc
      841 ccgcgtccca aaaagcgaag ggaggacggg gaggacggtg aggccagaag aagaaagcaa
      901 cagtaaaatc tcatgtgatt gcgtgcatgt catggatcta tgggatgcta gtccaccgtt
      961 tggaagtatg tgcagaagga gaaaggcgcg ggaggccaaa agcgacttcg gccacctctc
     1021 tactttctag aaagtttgct cgcagcttgt aataagcgcg gaaaaggtac tcgaaaaacc
     1081 ggagcaatcg gtgtggtatc agcagtcggg ttcgctaaac ctacgtcttc ttattttgga
     1141 atgttaaatt gaaagggcgt gccggcccta agcacggacg atgtgacaga cggaggagtg
     1201 tgagcaattc ggatgacgat gctggccggc agtcacgaca aagagtccga aggggcatgg
     1261 acagggcaat atataaatgg aaaagaggct ttgaatattc ctaacccttc gtcgtcggcc
     1321 tcccaacttg ccctcgcatg ctgaattggc cgcctatctg cgaatggccg gccgccgacc
     1381 ctaagtcgca ataacttgaa cggagccgat gcaaagtcct tatcgaacgc tcgttattat
     1441 aaacggggca gacctaggta tttaattata tcaaaaccct ccgcaatttc gctaatcatt
     1501 ccctaatttc ttctactttg gcgtctagtg ctgtggtcgc gcttgctctc aaagctcgct
     1561 gggaggcagt ggagcgtctg gcttaaaatc gatcgccgca accctagagt tcctttcact
     1621 tctttctctc ttgcgtggag agcatcgaca tcgaagagct gcatcttgtc ttctggtatg
     1681 cgccttgtgc cctcgtctct gtacgtcgtt acgattgctc gatattgtga ttgtctagac
     1741 ctgaaacccg actggagctt ttacgtggtg tcgattgggg cctcgagcga gtgatttttt
     1801 aggttcgttt tagccaagta catcccactt tggtcagtcc tgttatctcg gtaaggtgtt
     1861 cagttcggtc tctggtgcgg aggtgcctcg taagtttcct gcatgataga tttgtctatc
     1921 cttgacttga agaggagtcg aaggtcagcg aggttggctg tgtgctatgg acgtcggtgc
     1981 tccttggttc cgtgcttcta tgggtgttgc ctcaacagta ttgatgtgcc tgcagaaagg
     2041 ttgactagag ttatcaacaa gtttgtacaa aaaagcaggc tccgcggccg cccccttcac
     2101 catggataag aagtactcta tcggactcga tatcggaact aactctgtgg gatgggctgt
     2161 gatcaccgat gagtacaagg tgccatctaa gaagttcaag gttctcggaa acaccgatag
     2221 gcactctatc aagaaaaacc ttatcggtgc tctcctcttc gattctggtg aaactgctga
     2281 ggctaccaga ctcaagagaa ccgctagaag aaggtacacc agaagaaaga acaggatctg
     2341 ctacctccaa gagatcttct ctaacgagat ggctaaagtg gatgattcat tcttccacag
     2401 gctcgaagag tcattcctcg tggaagaaga taagaagcac gagaggcacc ctatcttcgg
     2461 aaacatcgtt gatgaggtgg cataccacga gaagtaccct actatctacc acctcagaaa
     2521 gaagctcgtt gattctactg ataaggctga tctcaggctc atctacctcg ctctcgctca
     2581 catgatcaag ttcagaggac acttcctcat cgagggtgat ctcaaccctg ataactctga
     2641 tgtggataag ttgttcatcc agctcgtgca gacctacaac cagcttttcg aagagaaccc
     2701 tatcaacgct tcaggtgtgg atgctaaggc tatcctctct gctaggctct ctaagtcaag
     2761 aaggcttgag aacctcattg ctcagctccc tggtgagaag aagaacggac ttttcggaaa
     2821 cttgatcgct ctctctctcg gactcacccc taacttcaag tctaacttcg atctcgctga
     2881 ggatgcaaag ctccagctct caaaggatac ctacgatgat gatctcgata acctcctcgc
     2941 tcagatcgga gatcagtacg ctgatttgtt cctcgctgct aagaacctct ctgatgctat
     3001 cctcctcagt gatatcctca gagtgaacac cgagatcacc aaggctccac tctcagcttc
     3061 tatgatcaag agatacgatg agcaccacca ggatctcaca cttctcaagg ctcttgttag
     3121 acagcagctc ccagagaagt acaaagagat tttcttcgat cagtctaaga acggatacgc
     3181 tggttacatc gatggtggtg catctcaaga agagttctac aagttcatca agcctatcct
     3241 cgagaagatg gatggaaccg aggaactcct cgtgaagctc aatagagagg atcttctcag
     3301 aaagcagagg accttcgata acggatctat ccctcatcag atccacctcg gagagttgca
     3361 cgctatcctt agaaggcaag aggatttcta cccattcctc aaggataaca gggaaaagat
     3421 tgagaagatt ctcaccttca gaatccctta ctacgtggga cctctcgcta gaggaaactc
     3481 aagattcgct tggatgacca gaaagtctga ggaaaccatc accccttgga acttcgaaga
     3541 ggtggtggat aagggtgcta gtgctcagtc tttcatcgag aggatgacca acttcgataa
     3601 gaaccttcca aacgagaagg tgctccctaa gcactctttg ctctacgagt acttcaccgt
     3661 gtacaacgag ttgaccaagg ttaagtacgt gaccgaggga atgaggaagc ctgctttttt
     3721 gtcaggtgag caaaagaagg ctatcgttga tctcttgttc aagaccaaca gaaaggtgac
     3781 cgtgaagcag ctcaaagagg attacttcaa gaaaatcgag tgcttcgatt cagttgagat
     3841 ttctggtgtt gaggataggt tcaacgcatc tctcggaacc taccacgatc tcctcaagat
     3901 cattaaggat aaggatttct tggataacga ggaaaacgag gatatcttgg aggatatcgt
     3961 tcttaccctc accctctttg aagatagaga gatgattgaa gaaaggctca agacctacgc
     4021 tcatctcttc gatgataagg tgatgaagca gttgaagaga agaagataca ctggttgggg
     4081 aaggctctca agaaagctca ttaacggaat cagggataag cagtctggaa agacaatcct
     4141 tgatttcctc aagtctgatg gattcgctaa cagaaacttc atgcagctca tccacgatga
     4201 ttctctcacc tttaaagagg atatccagaa ggctcaggtt tcaggacagg gtgatagtct
     4261 ccatgagcat atcgctaacc tcgctggatc tcctgcaatc aagaagggaa tcctccagac
     4321 tgtgaaggtt gtggatgagt tggtgaaggt gatgggaagg cataagcctg agaacatcgt
     4381 gatcgaaatg gctagagaga accagaccac tcagaaggga cagaagaact ctagggaaag
     4441 gatgaagagg atcgaggaag gtatcaaaga gcttggatct cagatcctca aagagcaccc
     4501 tgttgagaac actcagctcc agaatgagaa gctctacctc tactacctcc agaacggaag
     4561 ggatatgtat gtggatcaag agttggatat caacaggctc tctgattacg atgttgatca
     4621 tatcgtgcca cagtcattct tgaaggatga ttctatcgat aacaaggtgc tcaccaggtc
     4681 tgataagaac aggggtaaga gtgataacgt gccaagtgaa gaggttgtga agaaaatgaa
     4741 gaactattgg aggcagctcc tcaacgctaa gctcatcact cagagaaagt tcgataactt
     4801 gactaaggct gagaggggag gactctctga attggataag gcaggattca tcaagaggca
     4861 gcttgtggaa accaggcaga tcactaagca cgttgcacag atcctcgatt ctaggatgaa
     4921 caccaagtac gatgagaacg ataagttgat cagggaagtg aaggttatca ccctcaagtc
     4981 aaagctcgtg tctgatttca gaaaggattt ccaattctac aaggtgaggg aaatcaacaa
     5041 ctaccaccac gctcacgatg cttaccttaa cgctgttgtt ggaaccgctc tcatcaagaa
     5101 gtatcctaag ctcgagtcag agttcgtgta cggtgattac aaggtgtacg atgtgaggaa
     5161 gatgatcgct aagtctgagc aagagatcgg aaaggctacc gctaagtatt tcttctactc
     5221 taacatcatg aatttcttca agaccgagat taccctcgct aacggtgaga tcagaaagag
     5281 gccactcatc gagacaaacg gtgaaacagg tgagatcgtg tgggataagg gaagggattt
     5341 cgctaccgtt agaaaggtgc tctctatgcc acaggtgaac atcgttaaga aaaccgaggt
     5401 gcagaccggt ggattctcta aagagtctat cctccctaag aggaactctg ataagctcat
     5461 tgctaggaag aaggattggg accctaagaa atacggtggt ttcgattctc ctaccgtggc
     5521 ttactctgtt ctcgttgtgg ctaaggttga gaagggaaag agtaagaagc tcaagtctgt
     5581 taaggaactt ctcggaatca ctatcatgga aaggtcatct ttcgagaaga acccaatcga
     5641 tttcctcgag gctaagggat acaaagaggt taagaaggat ctcatcatca agctcccaaa
     5701 gtactcactc ttcgaactcg agaacggtag aaagaggatg ctcgcttctg ctggtgagct
     5761 tcaaaaggga aacgagcttg ctctcccatc taagtacgtt aactttcttt acctcgcttc
     5821 tcactacgag aagttgaagg gatctccaga agataacgag cagaagcaac ttttcgttga
     5881 gcagcacaag cactacttgg atgagatcat cgagcagatc tctgagttct ctaaaagggt
     5941 gatcctcgct gatgcaaacc tcgataaggt gttgtctgct tacaacaagc acagagataa
     6001 gcctatcagg gaacaggcag agaacatcat ccatctcttc acccttacca acctcggtgc
     6061 tcctgctgct ttcaagtact tcgatacaac catcgatagg aagagataca cctctaccaa
     6121 agaagtgctc gatgctaccc tcatccatca gtctatcact ggactctacg agactaggat
     6181 cgatctctca cagctcggtg gtgattcaag ggctgatcct aagaagaaga ggaaggtttg
     6241 aggcgcgccg agctccaggc ctcccagctt tcgtccgtat catcggtttc gacaacgttc
     6301 gtcaagttca atgcatcagt ttcattgccc acacaccaga atcctactaa gtttgagtat
     6361 tatggcattg gaaaagctgt tttcttctat catttgttct gcttgtaatt tactgtgttc
     6421 tttcagtttt tgttttcgga catcaaaatg caaatggatg gataagagtt aataaatgat
     6481 atggtccttt tgttcattct caaattatta ttatctgttg tttttacttt aatgggttga
     6541 atttaagtaa gaaaggaact aacagtgtga tattaaggtg caatgttaga catataaaac
     6601 agtctttcac ctctctttgg ttatgtcttg aattggtttg tttcttcact tatctgtgta
     6661 atcaagttta ctatgagtct atgatcaagt aattatgcaa tcaagttaag tacagtatag
     6721 gcttaagggt gggcgcgccg acccagcttt cttgtacaaa gtggttgata acagcggttg
     6781 actagagtta tcaacaagtt tgtacaaaaa agcaggctcc gcggccgccc ccttgtaacg
     6841 tgagactact acactcctca acgcaagttc tatctcatat ctctgcaaga gcttattcca
     6901 agaaactttc ttgtgcttct atgcatgccc aggtacctta agtcgcgcgc atagtcactt
     6961 cacagccttt tcagttctcc ctttgatggc tcagagtgac gcaactgcac aaaaataaga
     7021 aaaaagtcga agcgtcacga gagatgtatc cggctagttt ccttggagtt cggattgctc
     7081 tctttctctg tctccccgaa ccatatggaa tttatgaact gctgttcact agtcagtcac
     7141 ttgtttcaga acttttgagt ggtcgattta tccaccttct tgtttcttcc tgggtccaga
     7201 atctgacttc ttgagacaac aaacagtgca cagtggatcg aagaatgacc acagttgaga
     7261 cacgtggcgc tggcctcatt tattgcacca tacgtttagt gaagaactga ataatggtgg
     7321 atataaagca gagttgcacc cagcctctcg gaaacacacc ttcgggcgcc gttttagagc
     7381 tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt
     7441 cggtgctttt tttctagtat cgggaattct aaagggtggg cgcgccgacc cagctttctt
     7501 gtacaaagtg gttgataaca gcgcttagag gcttagagct cgaatttccc cgatcgttca
     7561 aacatttggc aataaagttt cttaagattg aatcctgttg ccggtcttgc gatgattatc
     7621 atataatttc tgttgaatta cgttaagcat gtaataatta acatgtaatg catgacgtta
     7681 tttatgagat gggtttttat gattagagtc ccgcaattat acatttaata cgcgatagaa
     7741 aacaaaatat agcgcgcaaa ctaggataaa ttatcgcgcg cggtgtcatc tatgttacta
     7801 gatcgggaat tggttccgga accaattcgt aatcatgtca tagctgtttc ctgtgtgaaa
     7861 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     7921 gggtgcctaa tgagtgagct aactcacatt aggctgaatt aggcgcgcct atttctgaat
     7981 tcaacacatt gcggacgttt ttaatgtact gaattaacgc cgaattaatt cgggggatct
     8041 ggattttagt actggatttt ggttttagga attagaaatt ttattgatag aagtatttta
     8101 caaatacaaa tacatactaa gggtttctta tatgctcaac acatgagcga aaccctatag
     8161 gaaccctaat tcccttatct gggaactact cacacattat tatggagaaa ctcgagcttg
     8221 tcgatcgaca gatcccggtc ggcatctact ctatttcttt gccctcggac gagtgctggg
     8281 gcgtcggttt ccactatcgg cgagtacttc tacacagcca tcggtccaga cggccgcgct
     8341 tctgcgggcg atttgtgtac gcccgacagt cccggctccg gatcggacga ttgcgtcgca
     8401 tcgaccctgc gcccaagctg catcatcgaa attgccgtca accaagctct gatagagttg
     8461 gtcaagacca atgcggagca tatacgcccg gagtcgtggc gatcctgcaa gctccggatg
     8521 cctccgctcg aagtagcgcg tctgctgctc catacaagcc aaccacggcc tccagaagaa
     8581 gatgttggcg acctcgtatt gggaatcccc gaacatcgcc tcgctccagt caatgaccgc
     8641 tgttatgcgg ccattgtccg tcaggacatt gttggagccg aaatccgcgt gcacgaggtg
     8701 ccggacttcg gggcagtcct cggcccaaag catcagctca tcgagagcct gcgcgacgga
     8761 cgcactgacg gtgtcgtcca tcacagtttg ccagtgatac acatggggat cagcaatcgc
     8821 gcatatgaaa tcacgccatg tagtgtattg accgattcct tgcggtccga atgggccgaa
     8881 cccgctcgtc tggctaagat cggccgcagc gatcgcatcc atagcctccg cgaccggttg
     8941 tagaacagcg ggcagttcgg tttcaggcag gtcttgcaac gtgacaccct gtgaacggcg
     9001 ggagatgcaa taggtcaggc tctcgctaaa ctccccaatg tcaagcactt ccggaatcgg
     9061 gagcgcggcc gatgcaaagt gccgataaac ataacgatct ttgtagaaac catcggcgca
     9121 gctatttacc cgcaggacat atccacgccc tcctacatcg aagctgaaag cacgagattc
     9181 ttcgccctcc gagagctgca tcaggtcgga gacgctgtcg aacttttcga tcagaaactt
     9241 ctcgacagac gtcgcggtga gttcaggctt tttcatatct cattgccccc cggatctgcg
     9301 aaagctcgag agagatagat ttgtagagag agactggtga tttcagcgtg tcctctccaa
     9361 atgaaatgaa cttccttata tagaggaagg gtcttgcgaa ggatagtggg attgtgcgtc
     9421 atcccttacg tcagtggaga tatcacatca atccacttgc tttgaagacg tggttggaac
     9481 gtcttctttt tccacgatgc tcctcgtggg tgggggtcca tctttgggac cactgtcggc
     9541 agaggcatct tgaacgatag cctttccttt atcgcaatga tggcatttgt aggtgccacc
     9601 ttccttttct actgtccttt tgatgaagtg acagatagct gggcaatgga atccgaggag
     9661 gtttcccgat attacccttt gttgaaaagt ctcaatagcc ctttggtctt ctgagactgt
     9721 atctttgata ttcttggagt agacgagagt gtcgtgctcc accatgttca catcaatcca
     9781 cttgctttga agacgtggtt ggaacgtctt ctttttccac gatgctcctc gtgggtgggg
     9841 gtccatcttt gggaccactg tcggcagagg catcttgaac gatagccttt cctttatcgc
     9901 aatgatggca tttgtaggtg ccaccttcct tttctactgt ccttttgatg aagtgacaga
     9961 tagctgggca atggaatccg aggaggtttc ccgatattac cctttgttga aaagtctcaa
    10021 tagccctttg gtcttctgag actgtatctt tgatattctt ggagtagacg agagtgtcgt
    10081 gctccaccat gttggcaagc tgctctagcc aatacgcaaa ccgaattcag aaataaattc
    10141 agcctaattc ggcgttaatt cagtacatta aaaacgtccg caatgtgtta ttaagttgtc
    10201 taagcgtcaa tttgtttaca ccacaatata tcctgccacc agccagccaa cagctccccg
    10261 accggcagct cggcacaaaa tcaccactcg atacaggcag cccatcagtc cgggacggcg
    10321 tcagcgggag agccgttgta aggcggcaga ctttgctcat gttaccgatg ctattcggaa
    10381 gaacggcaac taagctgccg ggtttgaaac acggatgatc tcgcggaggg tagcatgttg
    10441 attgtaacga tgacagagcg ttgctgcctg tgatcaattc gggcacgaac ccagtggaca
    10501 taagcctgtt cggttcgtaa gctgtaatgc aagtagcgta tgcgctcacg caactggtcc
    10561 agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat ggcttgttat
    10621 gactgttttt ttggggtaca gtctatgcct cgggcatcca agcagcaagc gcgttacgcc
    10681 gtgggtcgat gtttgatgtt atggagcagc aacgatgtta cgcagcaggg cagtcgccct
    10741 aaaacaaagt taaacatcat gggggaagcg gtgatcgccg aagtatcgac tcaactatca
    10801 gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt tgctggccgt acatttgtac
    10861 ggctccgcag tggatggcgg cctgaagcca cacagtgata ttgatttgct ggttacggtg
    10921 accgtaaggc ttgatgaaac aacgcggcga gctttgatca acgacctttt ggaaacttcg
    10981 gcttcccctg gagagagcga gattctccgc gctgtagaag tcaccattgt tgtgcacgac
    11041 gacatcattc cgtggcgtta tccagctaag cgcgaactgc aatttggaga atggcagcgc
    11101 aatgacattc ttgcaggtat cttcgagcca gccacgatcg acattgatct ggctatcttg
    11161 ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc cagcggcgga ggaactcttt
    11221 gatccggttc ctgaacagga tctatttgag gcgctaaatg aaaccttaac gctatggaac
    11281 tcgccgcccg actgggctgg cgatgagcga aatgtagtgc ttacgttgtc ccgcatttgg
    11341 tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg ctgccgactg ggcaatggag
    11401 cgcctgccgg cccagtatca gcccgtcata cttgaagcta gacaggctta tcttggacaa
    11461 gaagaagatc gcttggcctc gcgcgcagat cagttggaag aatttgtcca ctacgtgaaa
    11521 ggcgagatca ccaaggtagt cggcaaataa tgtctagcta gaaattcgtt caagccgacg
    11581 ccgcttcgcg gcgcggctta actcaagcgt tagatgcact aagcacataa ttgctcacag
    11641 ccaaactatc aggtcaagtc tgcttttatt atttttaagc gtgcataata agccctacac
    11701 aaattgggag atatatcatg catgaccaaa atcccttaac gtgagttttc gttccactga
    11761 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
    11821 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
    11881 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
    11941 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
    12001 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
    12061 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
    12121 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
    12181 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
    12241 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
    12301 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
    12361 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
    12421 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
    12481 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
    12541 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
    12601 tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag
    12661 ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc
    12721 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
    12781 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
    12841 gcgcgaggca gggtgccttg atgtgggcgc cggcggtcga gtggcgacgg cgcggcttgt
    12901 ccgcgccctg gtagattgcc tggccgtagg ccagccattt ttgagcggcc agcggccgcg
    12961 ataggccgac gcgaagcggc ggggcgtagg gagcgcagcg accgaagggt aggcgctttt
    13021 tgcagctctt cggctgtgcg ctggccagac agttatgcac aggccaggcg ggttttaaga
    13081 gttttaataa gttttaaaga gttttaggcg gaaaaatcgc cttttttctc ttttatatca
    13141 gtcacttaca tgtgtgaccg gttcccaatg tacggctttg ggttcccaat gtacgggttc
    13201 cggttcccaa tgtacggctt tgggttccca atgtacgtgc tatccacagg aaagagacct
    13261 tttcgacctt tttcccctgc tagggcaatt tgccctagca tctgctccgt acattaggaa
    13321 ccggcggatg cttcgccctc gatcaggttg cggtagcgca tgactaggat cgggccagcc
    13381 tgccccgcct cctccttcaa atcgtactcc ggcaggtcat ttgacccgat cagcttgcgc
    13441 acggtgaaac agaacttctt gaactctccg gcgctgccac tgcgttcgta gatcgtcttg
    13501 aacaaccatc tggcttctgc cttgcctgcg gcgcggcgtg ccaggcggta gagaaaacgg
    13561 ccgatgccgg gatcgatcaa aaagtaatcg gggtgaaccg tcagcacgtc cgggttcttg
    13621 ccttctgtga tctcgcggta catccaatca gctagctcga tctcgatgta ctccggccgc
    13681 ccggtttcgc tctttacgat cttgtagcgg ctaatcaagg cttcaccctc ggataccgtc
    13741 accaggcggc cgttcttggc cttcttcgta cgctgcatgg caacgtgcgt ggtgtttaac
    13801 cgaatgcagg tttctaccag gtcgtctttc tgctttccgc catcggctcg ccggcagaac
    13861 ttgagtacgt ccgcaacgtg tggacggaac acgcggccgg gcttgtctcc cttcccttcc
    13921 cggtatcggt tcatggattc ggttagatgg gaaaccgcca tcagtaccag gtcgtaatcc
    13981 cacacactgg ccatgccggc cggccctgcg gaaacctcta cgtgcccgtc tggaagctcg
    14041 tagcggatca cctcgccagc tcgtcggtca cgcttcgaca gacggaaaac ggccacgtcc
    14101 atgatgctgc gactatcgcg ggtgcccacg tcatagagca tcggaacgaa aaaatctggt
    14161 tgctcgtcgc ccttgggcgg cttcctaatc gacggcgcac cggctgccgg cggttgccgg
    14221 gattctttgc ggattcgatc agcggccgct tgccacgatt caccggggcg tgcttctgcc
    14281 tcgatgcgtt gccgctgggc ggcctgcgcg gccttcaact tctccaccag gtcatcaccc
    14341 agcgccgcgc cgatttgtac cgggccggat ggtttgcgac cgctcacgcc gattcctcgg
    14401 gcttgggggt tccagtgcca ttgcagggcc ggcagacaac ccagccgctt acgcctggcc
    14461 aaccgcccgt tcctccacac atggggcatt ccacggcgtc ggtgcctggt tgttcttgat
    14521 tttccatgcc gcctccttta gccgctaaaa ttcatctact catttattca tttgctcatt
    14581 tactctggta gctgcgcgat gtattcagat agcagctcgg taatggtctt gccttggcgt
    14641 accgcgtaca tcttcagctt ggtgtgatcc tccgccggca actgaaagtt gacccgcttc
    14701 atggctggcg tgtctgccag gctggccaac gttgcagcct tgctgctgcg tgcgctcgga
    14761 cggccggcac ttagcgtgtt tgtgcttttg ctcattttct ctttacctca ttaactcaaa
    14821 tgagttttga tttaatttca gcggccagcg cctggacctc gcgggcagcg tcgccctcgg
    14881 gttctgattc aagaacggtt gtgccggcgg cggcagtgcc tgggtagctc acgcgctgcg
    14941 tgatacggga ctcaagaatg ggcagctcgt acccggccag cgcctcggca acctcaccgc
    15001 cgatgcgcgt gcctttgatc gcccgcgaca cgacaaaggc cgcttgtagc cttccatccg
    15061 tgacctcaat gcgctgctta accagctcca ccaggtcggc ggtggcccat atgtcgtaag
    15121 ggcttggctg caccggaatc agcacgaagt cggctgcctt gatcgcggac acagccaagt
    15181 ccgccgcctg gggcgctccg tcgatcacta cgaagtcgcg ccggccgatg gccttcacgt
    15241 cgcggtcaat cgtcgggcgg tcgatgccga caacggttag cggttgatct tcccgcacgg
    15301 ccgcccaatc gcgggcactg ccctggggat cggaatcgac taacagaaca tcggccccgg
    15361 cgagttgcag ggcgcgggct agatgggttg cgatggtcgt cttgcctgac ccgcctttct
    15421 ggttaagtac agcgataacc ttcatgcgtt ccccttgcgt atttgtttat ttactcatcg
    15481 catcatatac gcagcgaccg catgacgcaa gctgttttac tcaaatacac atcacctttt
    15541 tagacggcgg cgctcggttt cttcagcggc caagctggcc ggccaggccg ccagcttggc
    15601 atcagacaaa ccggccagga tttcatgcag ccgcacggtt gagacgtgcg cgggcggctc
    15661 gaacacgtac ccggccgcga tcatctccgc ctcgatctct tcggtaatga aaaacggttc
    15721 gtcctggccg tcctggtgcg gtttcatgct tgttcctctt ggcgttcatt ctcggcggcc
    15781 gccagggcgt cggcctcggt caatgcgtcc tcacggaagg caccgcgccg cctggcctcg
    15841 gtgggcgtca cttcctcgct gcgctcaagt gcgcggtaca gggtcgagcg atgcacgcca
    15901 agcagtgcag ccgcctcttt cacggtgcgg ccttcctggt cgatcagctc gcgggcgtgc
    15961 gcgatctgtg ccggggtgag ggtagggcgg gggccaaact tcacgcctcg ggccttggcg
    16021 gcctcgcgcc cgctccgggt gcggtcgatg attagggaac gctcgaactc ggcaatgccg
    16081 gcgaacacgg tcaacaccat gcggccggcc ggcgtggtgg tgtcggccca cggctctgcc
    16141 aggctacgca ggcccgcgcc ggcctcctgg atgcgctcgg caatgtccag taggtcgcgg
    16201 gtgctgcggg ccaggcggtc tagcctggtc actgtcacaa cgtcgccagg gcgtaggtgg
    16261 tcaagcatcc tggccagctc cgggcggtcg cgcctggtgc cggtgatctt ctcggaaaac
    16321 agcttggtgc agccggccgc gtgcagttcg gcccgttggt tggtcaagtc ctggtcgtcg
    16381 gtgctgacgc gggcatagcc cagcaggcca gcggcggcgc tcttgttcat ggcgtaatgt
    16441 ctccggttct agtcgcaagt attctacttt atgcgactaa aacacgcgac aagaaaacgc
    16501 caggaaaagg gcagggcggc agcctgtcgc gtaacttagg acttgtgcga catgtcgttt
    16561 tcagaagacg gctgcactga acgtcagaag ccgactgcac tatagcagcg gaggggttgg
    16621 atcaaagtac tttgatcccg aggggaaccc tgtggttggc atgcacatac aaatggacga
    16681 acggataaac ct
//
