pKI1.1R

LOCUS       pKI1.1R               18557 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  CRISPR/Cas9 in Arabidopsis with seed a fluorescent reporter..
ACCESSION   .
VERSION     .
KEYWORDS    pKI1.1R
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 18557)
  AUTHORS   Tsutsui H, Higashiyama T
  TITLE     pKAMA-ITACHI vectors for highly efficient CRISPR/Cas9-mediated gene 
            knockout in Arabidopsis thaliana.
  JOURNAL   Plant Cell Physiol. 2016 Nov 17. pii: pcw191.
  PUBMED    27856772
REFERENCE   2  (bases 1 to 18557)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..18557
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     regulatory      19..28
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    41..65
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        548..623
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             2312..2377
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2384..2404
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2429..6529
                     /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             6530..6577
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     protein_bind    complement(6848..6872)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      9714..9965
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    10069..10093
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             11424..12053
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             12482..13555
                     /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      13621..13815
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(14051..14073)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    14159..14299
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     14214..14233
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(14314..14331)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(14485..15073)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(14565..14584)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(15319..16110)
                     /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"
     misc_feature    16631..16655
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      16748..17000
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(17198..18223)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(18259..18438)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
ORIGIN
        1 cgacgtcgca tgctcccggc cgccatggcc gcgggatatc acaagtttgt acaaaaaagc
       61 aggctccgcg gccgctcgtt gaacaacgga aactcgactt gccttccgca caatacatca
      121 tttcttctta gctttttttc ttcttcttcg ttcatacagt ttttttttgt ttatcagctt
      181 acattttctt gaaccgtagc tttcgttttc ttctttttaa ctttccattc ggagtttttg
      241 tatcttgttt catagtttgt cccaggatta gaatgattag gcatcgaacc ttcaagaatt
      301 tgattgaata aaacatcttc attcttaaga tatgaagata atcttcaaaa ggcccctggg
      361 aatctgaaag aagagaagca ggcccattta tatgggaaag aacaatagta tttcttatat
      421 aggcccattt aagttgaaaa caatcttcaa aagtcccaca tcgcttagat aagaaaacga
      481 agctgagttt atatacagct agagtcgaag tagtgattga aatgcaggtg atgactcacc
      541 tgcataagtt ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg
      601 aaaaagtggc accgagtcgg tgcttttttt ttttgcggcc gcgatccctc aacttttgat
      661 tcgctatttg cagtgcacct gtggcgttca tcacatcttt tgtgacactg tttgcactgg
      721 tcattgctat tacaaaggac cttcctgatg ttgaaggaga tcgaaagtaa gtaactgcac
      781 gcataaccat tttctttccg ctctttggct caatccattt gacagtcaaa gacaatgttt
      841 aaccagctcc gtttgatata ttgtctttat gtgtttgttc aagcatgttt agttaatcat
      901 gcctttgatt gatcttgaat aggttccaaa tatcaaccct ggcaacaaaa cttggagtga
      961 gaaacattgc attcctcggt tctggacttc tgctagtaaa ttatgtttca gccatatcac
     1021 tagctttcta catgcctcag gtgaattcat ctatttccgt cttaactatt tcggttaatt
     1081 aaagcacgaa caccattact gcatgtagaa gcttgataaa ctatcgccac caatttattt
     1141 ttgttgcgat attgttactt tcctcagtat gcagctttga aaagaccaac cctcttatcc
     1201 tttaacaatg aacaggtttt tagaggtagc ttgatgattc ctgcacatgt gatcttggct
     1261 tcaggcttaa ttttccaggt aaagcattat gagatactct tatatctctt acatactttt
     1321 gagataatgc acaagaactt cataactata tgctttagtt tctgcatttg acactgccaa
     1381 attcattaat ctctaatatc tttgttgttg atctttggta gacatgggta ctagaaaaag
     1441 caaactacac caaggtaaaa tacttttgta caaacataaa ctcgttatca cggaacatca
     1501 atggagtgta tatctaacgg agtgtagaaa catttgatta ttgcaggaag ctatctcagg
     1561 atattatcgg tttatatgga atctcttcta cgcagagtat ctgttattcc ccttcctcta
     1621 gctttcaatt tcatggtgag gatatgcagt tttctttgta tatcattctt cttcttcttt
     1681 gtagcttgga gtcaaaatcg gttccttcat gtacatacat caaggatatg tccttctgaa
     1741 tttttatatc ttgcaataaa aatgcttgta ccaattgaaa caccagcttt ttgagttcta
     1801 tgatcactga cttggttcta accaaaaaaa aaaaatgttt aatttacata tctaaaagta
     1861 ggtttaggga aacctaaaca gtaaaatatt tgtatattat tcgaatttca ctcatcataa
     1921 aaacttaaat tgcaccataa aattttgttt tactattaat gatgtaattt gtgtaactta
     1981 agataaaaat aatattccgt aagttaaccg gctaaaacca cgtataaacc agggaacctg
     2041 ttaaaccggt tctttactgg ataaagaaat gaaagcccat gtagacagct ccattagagc
     2101 ccaaacccta aatttctcat ctatataaaa ggagtgacat tagggttttt gttcgtcctc
     2161 ttaaagcttc tcgttttctc tgccgtctct ctcattcgcg cgacgcaaac gatcttcagg
     2221 tgatcttctt tctccaaatc ctctctcata actctgattt cgtacttgtg tatttgagct
     2281 cacgctctgt ttctctcacc acagccccat ggactataag gaccacgacg gagactacaa
     2341 ggatcatgat attgattaca aagacgatga cgataagatg gccccaaaga agaagcggaa
     2401 ggtcggtatc cacggagtcc cagcagccga caagaagtac agcatcggcc tggacatcgg
     2461 caccaactct gtgggctggg ccgtgatcac cgacgagtac aaggtgccca gcaagaaatt
     2521 caaggtgctg ggcaacaccg accggcacag catcaagaag aacctgatcg gagccctgct
     2581 gttcgacagc ggcgaaacag ccgaggccac ccggctgaag agaaccgcca gaagaagata
     2641 caccagacgg aagaaccgga tctgctatct gcaagagatc ttcagcaacg agatggccaa
     2701 ggtggacgac agcttcttcc acagactgga agagtccttc ctggtggaag aggataagaa
     2761 gcacgagcgg caccccatct tcggcaacat cgtggacgag gtggcctacc acgagaagta
     2821 ccccaccatc taccacctga gaaagaaact ggtggacagc accgacaagg ccgacctgcg
     2881 gctgatctat ctggccctgg cccacatgat caagttccgg ggccacttcc tgatcgaggg
     2941 cgacctgaac cccgacaaca gcgacgtgga caagctgttc atccagctgg tgcagaccta
     3001 caaccagctg ttcgaggaaa accccatcaa cgccagcggc gtggacgcca aggccatcct
     3061 gtctgccaga ctgagcaaga gcagacggct ggaaaatctg atcgcccagc tgcccggcga
     3121 gaagaagaat ggcctgttcg gaaacctgat tgccctgagc ctgggcctga cccccaactt
     3181 caagagcaac ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg acacctacga
     3241 cgacgacctg gacaacctgc tggcccagat cggcgaccag tacgccgacc tgtttctggc
     3301 cgccaagaac ctgtccgacg ccatcctgct gagcgacatc ctgagagtga acaccgagat
     3361 caccaaggcc cccctgagcg cctctatgat caagagatac gacgagcacc accaggacct
     3421 gaccctgctg aaagctctcg tgcggcagca gctgcctgag aagtacaaag agattttctt
     3481 cgaccagagc aagaacggct acgccggcta cattgacggc ggagccagcc aggaagagtt
     3541 ctacaagttc atcaagccca tcctggaaaa gatggacggc accgaggaac tgctcgtgaa
     3601 gctgaacaga gaggacctgc tgcggaagca gcggaccttc gacaacggca gcatccccca
     3661 ccagatccac ctgggagagc tgcacgccat tctgcggcgg caggaagatt tttacccatt
     3721 cctgaaggac aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc cctactacgt
     3781 gggccctctg gccaggggaa acagcagatt cgcctggatg accagaaaga gcgaggaaac
     3841 catcaccccc tggaacttcg aggaagtggt ggacaagggc gcttccgccc agagcttcat
     3901 cgagcggatg accaacttcg ataagaacct gcccaacgag aaggtgctgc ccaagcacag
     3961 cctgctgtac gagtacttca ccgtgtataa cgagctgacc aaagtgaaat acgtgaccga
     4021 gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg tggacctgct
     4081 gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa gaggactact tcaagaaaat
     4141 cgagtgcttc gactccgtgg aaatctccgg cgtggaagat cggttcaacg cctccctggg
     4201 cacataccac gatctgctga aaattatcaa ggacaaggac ttcctggaca atgaggaaaa
     4261 cgaggacatt ctggaagata tcgtgctgac cctgacactg tttgaggaca gagagatgat
     4321 cgaggaacgg ctgaaaacct atgcccacct gttcgacgac aaagtgatga agcagctgaa
     4381 gcggcggaga tacaccggct ggggcaggct gagccggaag ctgatcaacg gcatccggga
     4441 caagcagtcc ggcaagacaa tcctggattt cctgaagtcc gacggcttcg ccaacagaaa
     4501 cttcatgcag ctgatccacg acgacagcct gacctttaaa gaggacatcc agaaagccca
     4561 ggtgtccggc cagggcgata gcctgcacga gcacattgcc aatctggccg gcagccccgc
     4621 cattaagaag ggcatcctgc agacagtgaa ggtggtggac gagctcgtga aagtgatggg
     4681 ccggcacaag cccgagaaca tcgtgatcga aatggccaga gagaaccaga ccacccagaa
     4741 gggacagaag aacagccgcg agagaatgaa gcggatcgaa gagggcatca aagagctggg
     4801 cagccagatc ctgaaagaac accccgtgga aaacacccag ctgcagaacg agaagctgta
     4861 cctgtactac ctgcagaatg ggcgggatat gtacgtggac caggaactgg acatcaaccg
     4921 gctgtccgac tacgatgtgg accatatcgt gcctcagagc tttctgaagg acgactccat
     4981 cgacaacaag gtgctgacca gaagcgacaa gaaccggggc aagagcgaca acgtgccctc
     5041 cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg ccaagctgat
     5101 tacccagaga aagttcgaca atctgaccaa ggccgagaga ggcggcctga gcgaactgga
     5161 taaggccggc ttcatcaaga gacagctggt ggaaacccgg cagatcacaa agcacgtggc
     5221 acagatcctg gactcccgga tgaacactaa gtacgacgag aatgacaagc tgatccggga
     5281 agtgaaagtg atcaccctga agtccaagct ggtgtccgat ttccggaagg atttccagtt
     5341 ttacaaagtg cgcgagatca acaactacca ccacgcccac gacgcctacc tgaacgccgt
     5401 cgtgggaacc gccctgatca aaaagtaccc taagctggaa agcgagttcg tgtacggcga
     5461 ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc
     5521 taccgccaag tacttcttct acagcaacat catgaacttt ttcaagaccg agattaccct
     5581 ggccaacggc gagatccgga agcggcctct gatcgagaca aacggcgaaa ccggggagat
     5641 cgtgtgggat aagggccggg attttgccac cgtgcggaaa gtgctgagca tgccccaagt
     5701 gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc agcaaagagt ctatcctgcc
     5761 caagaggaac agcgataagc tgatcgccag aaagaaggac tgggacccta agaagtacgg
     5821 cggcttcgac agccccaccg tggcctattc tgtgctggtg gtggccaaag tggaaaaggg
     5881 caagtccaag aaactgaaga gtgtgaaaga gctgctgggg atcaccatca tggaaagaag
     5941 cagcttcgag aagaatccca tcgactttct ggaagccaag ggctacaaag aagtgaaaaa
     6001 ggacctgatc atcaagctgc ctaagtactc cctgttcgag ctggaaaacg gccggaagag
     6061 aatgctggcc tctgccggcg aactgcagaa gggaaacgaa ctggccctgc cctccaaata
     6121 tgtgaacttc ctgtacctgg ccagccacta tgagaagctg aagggctccc ccgaggataa
     6181 tgagcagaaa cagctgtttg tggaacagca caagcactac ctggacgaga tcatcgagca
     6241 gatcagcgag ttctccaaga gagtgatcct ggccgacgct aatctggaca aagtgctgtc
     6301 cgcctacaac aagcaccggg ataagcccat cagagagcag gccgagaata tcatccacct
     6361 gtttaccctg accaatctgg gagcccctgc cgccttcaag tactttgaca ccaccatcga
     6421 ccggaagagg tacaccagca ccaaagaggt gctggacgcc accctgatcc accagagcat
     6481 caccggcctg tacgagacac ggatcgacct gtctcagctg ggaggcgaca aaaggccggc
     6541 ggccacgaaa aaggccggcc aggcaaaaaa gaaaaagtaa aagggtggga tatgaagatg
     6601 aagatgaaat atttggtgtg tcaaataaaa agcttgtgtg cttaagtttg tgtttttttc
     6661 ttggcttgtt gtgttatgaa tttgtggctt tttctaatat taaatgaatg taagatctca
     6721 ttataatgaa taaacaaatg tttctataat ccattgtgaa tgttttgttg gatctcttct
     6781 gcagcatata actactgtat gtgctatggt atggactatg gaatatgatt aaagataagg
     6841 cgcgccgacc cagctttctt gtacaaagtg gtgatatcac tagtgtatgt aggtatagta
     6901 acatgaacaa gaacagactc aagtacaaga tcgcatacga aaatggaaat ggcaatgtca
     6961 cttccacata atcaaacacg aatcctcata tcaacaaggc ctgagattct aactagctca
     7021 taacaactta gccaatagtt acttgagact accaaatgta tgtagaacta aagactaagg
     7081 gacagagagt tcgtctaaac aggtgaatct agtcgttgtt atctaataaa caattcagcc
     7141 ccaaatgcag aacacacata gagctctcta ttgattcaaa ttacgatctg atactgataa
     7201 cgtctagatt tttagggtta aagcaatcaa tcacctgacg attcaaggtg gttggatcat
     7261 gacgattcca gaaaacatca agcaagctct caaagctaca ctctttggga tcatactgaa
     7321 ctctaacaac ctcgttatgt cccgtagtgc cagtacagac atcctcgtaa ctcggattgt
     7381 gcacgatgcc atggctatac ccaacctcgg tcttggtcac accaggaact ctctggtaag
     7441 ctagctccac tccccagaaa caaccggcgc caaattgcgc gaattgctga cctgaagacg
     7501 gaacatcatc gtcgggtcct tgggcgattg cggcggaaga tgggtcagct tgggcttgag
     7561 gacgagaccc gaatccgagt ctgttgaaaa ggttgttcat tggggatttg tatacggaga
     7621 ttggtcgtcg agaggtttga gggaaaggac aaatgggttt ggctctggag aaagagagtg
     7681 cggctttaga gagagaattg agaggtttag agagagatgc ggcggcgatg agcggaggag
     7741 agacgacgag gacctgcatt atcaaagcag tgacgtggtg aaatttggaa cttttaagag
     7801 gcagatagat ttattatttg tatccatttt cttcattgtt ctagaatgtc gcggaacaaa
     7861 ttttaaaact aaatcctaaa tttttctaat tttgttgcca atagtggata tgtgggccgt
     7921 atagaaggaa tctattgaag gcccaaaccc atactgacga gcccaaaggt tcgttttgcg
     7981 ttttatgttt cggttcgatg ccaacgccac attctgagct aggcaaaaaa caaacgtgtc
     8041 tttgaataga ctcctctcgt taacacatgc agcggctgca tggtgacgcc attaacacgt
     8101 ggcctacaat tgcatgatgt ctccattgac acgtgacttc tcgtctcctt tcttaatata
     8161 tctaacaaac actcctacct cttccaaaat atatacacat ctttttgatc aatctctcat
     8221 tcaaaatctc attctctcta gtaaacaaga acaaaaaaat ggcggataca gctagaggaa
     8281 cccatcacga tatcatcggc agagaccagt acccgatgat gggccgagac cgagaccagt
     8341 accagatgtc cggacgagga tctgactact ccaagtctag gcagattgct aaagctgcaa
     8401 ctgctgtcac agctggtggt tccctccttg ttctctccag ccttaccctt gttggaactg
     8461 tcatagcttt gactgttgca acacctctgc tcgttatctt cagcccaatc cttgtcccgg
     8521 ctctcatcac agttgcactc ctcatcaccg gttttctttc ctctggaggg tttggcattg
     8581 ccgctataac cgttttctct tggatttaca agtaagcaca catttatcat cttacttcat
     8641 aattttgtgc aatatgtgca tgcatgtgtt gagccagtag ctttggatca atttttttgg
     8701 tcgaataaca aatgtaacaa taagaaattg caaattctag ggaacatttg gttaactaaa
     8761 tacgaaattt gacctagcta gcttgaatgt gtctgtgtat atcatctata taggtaaaat
     8821 gcttggtatg atacctattg attgtgaata ggtacgcaac gggagagcac ccacagggat
     8881 cagacaagtt ggacagtgca aggatgaagt tgggaagcaa agctcaggat ctgaaagaca
     8941 gagctcagta ctacggacag caacatactg gtggggaaca tgaccgtgac cgtactcgtg
     9001 gtggccagca cactactatg agcgagctga ttaaggagaa catgcacatg aagctgtaca
     9061 tggagggcac cgtgaacaac caccacttca agtgcacatc cgagggcgaa ggcaagccct
     9121 acgagggcac ccagaccatg agaatcaagg tggtcgaggg cggccctctc cccttcgcct
     9181 tcgacatcct ggctaccagc ttcatgtacg gcagcagaac cttcatcaac cacacccagg
     9241 gcatccccga cttctttaag cagtccttcc ctgagggctt cacatgggag agagtcacca
     9301 catacgaaga cgggggcgtg ctgaccgcta cccaggacac cagcctccag gacggctgcc
     9361 tcatctacaa cgtcaagatc agaggggtga acttcccatc caacggccct gtgatgcaga
     9421 agaaaacact cggctgggag gccaacaccg agatgctgta ccccgctgac ggcggcctgg
     9481 aaggcagaag cgacatggcc ctgaagctcg tgggcggggg ccacctgatc tgcaacttca
     9541 agaccacata cagatccaag aaacccgcta agaacctcaa gatgcccggc gtctactatg
     9601 tggaccacag actggaaaga atcaaggagg ccgacaaaga gacctacgtc gagcagcacg
     9661 aggtggctgt ggccagatac tgcgacctcc ctagcaaact ggggcacaag tgaatcgttc
     9721 aaacatttgg caataaagtt tcttaagatt gaatcctgtt gccggtcttg cgatgattat
     9781 catataattt ctgttgaatt acgttaagca tgtaataatt aacatgtaat gcatgacgtt
     9841 atttatgaga tgggttttta tgattagagt cccgcaatta tacatttaat acgcgataga
     9901 aaacaaaata tagcgcgcaa actaggataa attatcgcgc gcggtgtcat ctatgttact
     9961 agatcactag tgcggccgcc tgcaggtcga ccatatggga gagctcaagc ttagcttgag
    10021 cttggatcag attgtcgttt cccgccttca gtttaaacta tcagtgtttg acaggatata
    10081 ttggcgggta aacctaagag aaaagagcgt ttattagaat aatcggatat ttaaaagggc
    10141 gtgaaaaggt ttatccgttc gtccatttgt atgtgcatgc caaccacagg gttcccctcg
    10201 ggatcaaagt actttaaagt actttaaagt actttaaagt actttgatcc aacccctccg
    10261 ctgctatagt gcagtcggct tctgacgttc agtgcagccg tcttctgaaa acgacatgtc
    10321 gcacaagtcc taagttacgc gacaggctgc cgccctgccc ttttcctggc gttttcttgt
    10381 cgcgtgtttt agtcgcataa agtagaatac ttgcgactag aaccggagac attacgccat
    10441 gaacaagagc gccgccgctg gcctgctggg ctatgcccgc gtcagcaccg acgaccagga
    10501 cttgaccaac caacgggccg aactgcacgc ggccggctgc accaagctgt tttccgagaa
    10561 gatcaccggc accaggcgcg accgcccgga gctggccagg atgcttgacc acctacgccc
    10621 tggcgacgtt gtgacagtga ccaggctaga ccgcctggcc cgcagcaccc gcgacctact
    10681 ggacattgcc gagcgcatcc aggaggccgg cgcgggcctg cgtagcctgg cagagccgtg
    10741 ggccgacacc accacgccgg ccggccgcat ggtgttgacc gtgttcgccg gcattgccga
    10801 gttcgagcgt tccctaatca tcgaccgcac ccggagcggg cgcgaggccg ccaaggcccg
    10861 aggcgtgaag tttggccccc gccctaccct caccccggca cagatcgcgc acgcccgcga
    10921 gctgatcgac caggaaggcc gcaccgtgaa agaggcggct gcactgcttg gcgtgcatcg
    10981 ctcgaccctg taccgcgcac ttgagcgcag cgaggaagtg acgcccaccg aggccaggcg
    11041 gcgcggtgcc ttccgtgagg acgcattgac cgaggccgac gccctggcgg ccgccgagaa
    11101 tgaacgccaa gaggaacaag catgaaaccg caccaggacg gccaggacga accgtttttc
    11161 attaccgaag agatcgaggc ggagatgatc gcggccgggt acgtgttcga gccgcccgcg
    11221 cacgtctcaa ccgtgcggct gcatgaaatc ctggccggtt tgtctgatgc caagctggcg
    11281 gcctggccgg ccagcttggc cgctgaagaa accgagcgcc gccgtctaaa aaggtgatgt
    11341 gtatttgagt aaaacagctt gcgtcatgcg gtcgctgcgt atatgatgcg atgagtaaat
    11401 aaacaaatac gcaaggggaa cgcatgaagg ttatcgctgt acttaaccag aaaggcgggt
    11461 caggcaagac gaccatcgca acccatctag cccgcgccct gcaactcgcc ggggccgatg
    11521 ttctgttagt cgattccgat ccccagggca gtgcccgcga ttgggcggcc gtgcgggaag
    11581 atcaaccgct aaccgttgtc ggcatcgacc gcccgacgat tgaccgcgac gtgaaggcca
    11641 tcggccggcg cgacttcgta gtgatcgacg gagcgcccca ggcggcggac ttggctgtgt
    11701 ccgcgatcaa ggcagccgac ttcgtgctga ttccggtgca gccaagccct tacgacatat
    11761 gggccaccgc cgacctggtg gagctggtta agcagcgcat tgaggtcacg gatggaaggc
    11821 tacaagcggc ctttgtcgtg tcgcgggcga tcaaaggcac gcgcatcggc ggtgaggttg
    11881 ccgaggcgct ggccgggtac gagctgccca ttcttgagtc ccgtatcacg cagcgcgtga
    11941 gctacccagg cactgccgcc gccggcacaa ccgttcttga atcagaaccc gagggcgacg
    12001 ctgcccgcga ggtccaggcg ctggccgctg aaattaaatc aaaactcatt tgagttaatg
    12061 aggtaaagag aaaatgagca aaagcacaaa cacgctaagt gccggccgtc cgagcgcacg
    12121 cagcagcaag gctgcaacgt tggccagcct ggcagacacg ccagccatga agcgggtcaa
    12181 ctttcagttg ccggcggagg atcacaccaa gctgaagatg tacgcggtac gccaaggcaa
    12241 gaccattacc gagctgctat ctgaatacat cgcgcagcta ccagagtaaa tgagcaaatg
    12301 aataaatgag tagatgaatt ttagcggcta aaggaggcgg catggaaaat caagaacaac
    12361 caggcaccga cgccgtggaa tgccccatgt gtggaggaac gggcggttgg ccaggcgtaa
    12421 gcggctgggt tgtctgccgg ccctgcaatg gcactggaac ccccaagccc gaggaatcgg
    12481 cgtgagcggt cgcaaaccat ccggcccggt acaaatcggc gcggcgctgg gtgatgacct
    12541 ggtggagaag ttgaaggccg cgcaggccgc ccagcggcaa cgcatcgagg cagaagcacg
    12601 ccccggtgaa tcgtggcaag cggccgctga tcgaatccgc aaagaatccc ggcaaccgcc
    12661 ggcagccggt gcgccgtcga ttaggaagcc gcccaagggc gacgagcaac cagatttttt
    12721 cgttccgatg ctctatgacg tgggcacccg cgatagtcgc agcatcatgg acgtggccgt
    12781 tttccgtctg tcgaagcgtg accgacgagc tggcgaggtg atccgctacg agcttccaga
    12841 cgggcacgta gaggtttccg cagggccggc cggcatggcc agtgtgtggg attacgacct
    12901 ggtactgatg gcggtttccc atctaaccga atccatgaac cgataccggg aagggaaggg
    12961 agacaagccc ggccgcgtgt tccgtccaca cgttgcggac gtactcaagt tctgccggcg
    13021 agccgatggc ggaaagcaga aagacgacct ggtagaaacc tgcattcggt taaacaccac
    13081 gcacgttgcc atgcagcgta cgaagaaggc caagaacggc cgcctggtga cggtatccga
    13141 gggtgaagcc ttgattagcc gctacaagat cgtaaagagc gaaaccgggc ggccggagta
    13201 catcgagatc gagctagctg attggatgta ccgcgagatc acagaaggca agaacccgga
    13261 cgtgctgacg gttcaccccg attacttttt gatcgatccc ggcatcggcc gttttctcta
    13321 ccgcctggca cgccgcgccg caggcaaggc agaagccaga tggttgttca agacgatcta
    13381 cgaacgcagt ggcagcgccg gagagttcaa gaagttctgt ttcaccgtgc gcaagctgat
    13441 cgggtcaaat gacctgccgg agtacgattt gaaggaggag gcggggcagg ctggcccgat
    13501 cctagtcatg cgctaccgca acctgatcga gggcgaagca tccgccggtt cctaatgtac
    13561 ggagcagatg ctagggcaaa ttgccctagc aggggaaaaa ggtcgaaaag gtctctttcc
    13621 tgtggatagc acgtacattg ggaacccaaa gccgtacatt gggaaccgga acccgtacat
    13681 tgggaaccca aagccgtaca ttgggaaccg gtcacacatg taagtgactg atataaaaga
    13741 gaaaaaaggc gatttttccg cctaaaactc tttaaaactt attaaaactc ttaaaacccg
    13801 cctggcctgt gcataactgt ctggccagcg cacagccgaa gagctgcaaa aagcgcctac
    13861 ccttcggtcg ctgcgctccc tacgccccgc cgcttcgcgt cggcctatcg cggccgctgg
    13921 ccgctcaaaa atggctggcc tacggccagg caatctacca gggcgcggac aagccgcgcc
    13981 gtcgccactc gaccgccggc gcccacatca aggcaccctg cctcgcgcgt ttcggtgatg
    14041 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
    14101 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg
    14161 cagccatgac ccagtcacgt agcgatagcg gagtgtatac tggcttaact atgcggcatc
    14221 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
    14281 gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
    14341 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
    14401 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
    14461 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
    14521 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
    14581 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
    14641 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
    14701 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
    14761 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
    14821 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
    14881 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
    14941 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
    15001 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
    15061 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
    15121 aaactcacgt taagggattt tggtcatgca tgatatatct cccaatttgt gtagggctta
    15181 ttatgcacgc ttaaaaataa taaaagcaga cttgacctga tagtttggct gtgagcaatt
    15241 atgtgcttag tgcatctaac gcttgagtta agccgcgccg cgaagcggcg tcggcttgaa
    15301 cgaatttcta gctagacatt atttgccgac taccttggtg atctcgcctt tcacgtagtg
    15361 gacaaattct tccaactgat ctgcgcgcga ggccaagcga tcttcttctt gtccaagata
    15421 agcctgtcta gcttcaagta tgacgggctg atactgggcc ggcaggcgct ccattgccca
    15481 gtcggcagcg acatccttcg gcgcgatttt gccggttact gcgctgtacc aaatgcggga
    15541 caacgtaagc actacatttc gctcatcgcc agcccagtcg ggcggcgagt tccatagcgt
    15601 taaggtttca tttagcgcct caaatagatc ctgttcagga accggatcaa agagttcctc
    15661 cgccgctgga cctaccaagg caacgctatg ttctcttgct tttgtcagca agatagccag
    15721 atcaatgtcg atcgtggctg gctcgaagat acctgcaaga atgtcattgc gctgccattc
    15781 tccaaattgc agttcgcgct tagctggata acgccacgga atgatgtcgt cgtgcacaac
    15841 aatggtgact tctacagcgc ggagaatctc gctctctcca ggggaagccg aagtttccaa
    15901 aaggtcgttg atcaaagctc gccgcgttgt ttcatcaagc cttacggtca ccgtaaccag
    15961 caaatcaata tcactgtgtg gcttcaggcc gccatccact gcggagccgt acaaatgtac
    16021 ggccagcaac gtcggttcga gatggcgctc gatgacgcca actacctctg atagttgagt
    16081 cgatacttcg gcgatcaccg cttcccccat gatgtttaac tttgttttag ggcgactgcc
    16141 ctgctgcgta acatcgttgc tgctccataa catcaaacat cgacccacgg cgtaacgcgc
    16201 ttgctgcttg gatgcccgag gcatagactg taccccaaaa aaacagtcat aacaagccat
    16261 gaaaaccgcc actgcgccgt taccaccgct gcgttcggtc aaggttctgg accagttgcg
    16321 tgagcgcata cgctacttgc attacagctt acgaaccgaa caggcttatg tccactgggt
    16381 tcgtgcccga attgatcaca ggcagcaacg ctctgtcatc gttacaatca acatgctacc
    16441 ctccgcgaga tcatccgtgt ttcaaacccg gcagcttagt tgccgttctt ccgaatagca
    16501 tcggtaacat gagcaaagtc tgccgcctta caacggctct cccgctgacg ccgtcccgga
    16561 ctgatgggct gcctgtatcg agtggtgatt ttgtgccgag ctgccggtcg gggagctgtt
    16621 ggctggctgg tggcaggata tattgtggtg taaacaaatt gacgcttaga caacttaata
    16681 acacattgcg gacgttttta atgtactgaa ttaacgccga attgaattat cagcttgcat
    16741 gccggtcgat ctagtaacat agatgacacc gcgcgcgata atttatccta gtttgcgcgc
    16801 tatattttgt tttctatcgc gtattaaatg tataattgcg ggactctaat cataaaaacc
    16861 catctcataa ataacgtcat gcattacatg ttaattatta catgcttaac gtaattcaac
    16921 agaaattata tgataatcat cgcaagaccg gcaacaggat tcaatcttaa gaaactttat
    16981 tgccaaatgt ttgaacgatc tgcttgactc tagggaatta attcctgaat cactgcgacc
    17041 ggccctcccg cgacccagcc gagcgagctt agcgaactgt ggacgagaac tgtgccacca
    17101 agcgtaaggc cgttctctcg catttgcctt gctaggctcg cgcgagttgc tggctgaggc
    17161 gttctcgaaa tcagctcttg ttcggtcggc atctactcta ttcctttgcc ctcggacgag
    17221 tgctggggcg tcggtttcca ctatcggcga gtacttctac acagccatcg gtccagacgg
    17281 ccgcgcttct gcgggcgatt tgtgtacgcc cgacagtccc ggctccggat cggacgattg
    17341 cgtcgcatcg accctgcgcc caagctgcat catcgaaatt gccgtcaacc aagctctgat
    17401 agagttggtc aagaccaatg cggagcatat acgcccggag ccttggcgat cctgcaagct
    17461 ccggatgcct ccgctcgaag tagcgcgtct gctgctccat acaagccaac cacggcctcc
    17521 agaagaagat gttggcgacc tcgtattggg aatccccgaa catcgcctcg ctccagtcaa
    17581 tgaccgctgt tatgcggcca ttgtccgtca ggacattgtt ggagccgaaa tccgcgtgca
    17641 cgaggtgccg gacttcgggg cagtcctcgg cccaaagcat cagctcatcg agagcctgcg
    17701 cgacggacgc actgacggtg tcgtccatca cagtttgcca gtgatacaca tggggatcag
    17761 caatcgcgca tatgaaatca cgccatgtag tgtattgacc gattccttgc ggtccgaatg
    17821 ggccgaaccc gctcgtctgg ctaagatcgg ccgcagcgat agcatccata gcctccgcga
    17881 ccggctgaag aacagcgggc agttcggttt caggcaggtc ttgcaacgtg acaccctgtg
    17941 cacggcggga gatgcaatag gtcaggctct cgctgaactc cccaatgtca agcacttccg
    18001 gaatcgggag cgcggccgat gcaaagtgcc gataaacata acgatctttg tagaaaccat
    18061 cggcgcagct atttacccgc aggacatatc cacgccctcc tacatcgaag ctgaaagcac
    18121 gagattcttc gccctccgag agctgcatca ggtcggagac gctgtcgaac ttttcgatca
    18181 gaaacttctc gacagacgtc gcggtgagtt caggcttttt catagggggg attcgagttg
    18241 agagtgaata tgagactcta attggatacc gaggggaatt tatggaacgt cagtggagca
    18301 tttttgacaa gaaatatttg ctagctgata gtgaccttag gcgacttttg aacgcgcaat
    18361 aatggtttct gacgtatgtg cttagctcat taaactccag aaacccgcgg ctcagtggct
    18421 ccttcaacgt tgcggttctg tcagttccaa acgtaaaacg gcttgtcccg cgtcatcggc
    18481 gggggtcata acgtgactcc cttaattctc cgctcatgat cgataattcg cggtacccgg
    18541 ggatcctcta gagggcc
//