pRGEB31

LOCUS       pRGEB31            15037 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  Binary vector derived from pRGE31. Deliver sgRNA and Cas9 into 
            plant. by agrobacterium mediated transformation..
ACCESSION   .
VERSION     .
KEYWORDS    pRGEB31
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15037)
  AUTHORS   Xie K, Yang Y
  TITLE     RNA-Guided Genome Editing in Plants Using a CRISPR-Cas System.
  JOURNAL   Mol Plant. 2013 Nov;6(6):1975-83. doi: 10.1093/mp/sst119. Epub 2013 
            Aug 17.
  PUBMED    23956122
REFERENCE   2  (bases 1 to 15037)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..15037
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        6..386
                     /label=OsU3 promoter
                     /note="Oryza sativa (rice) snRNA U3 promoter"
     misc_RNA        403..478
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        1044..1389
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     1332..1352
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     protein_bind    1411..1435
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1480..1545
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1552..1572
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1597..5697
                     /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             5698..5745
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     protein_bind    complement(5778..5802)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5813..5836
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      5858..6110
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(6119..6135)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6136..6152)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6145..6167)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    6355..6379
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             7679..8308
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             8742..9809
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     rep_origin      9875..10069
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(10305..10327)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    10413..10553
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     10468..10487
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(10568..10585)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10739..11327)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10819..10838)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(11414..12208)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    12633..12657
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    12735..12909
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(12949..13974)
                     /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
                     K"
     promoter        complement(14042..14719)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     protein_bind    14910..14931
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        14946..14976
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    14984..15000
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     14989..15011
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     15008..15024
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     15008..15024
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 agcttaagga atctttaaac atacgaacag atcacttaaa gttcttctga agcaacttaa
       61 agttatcagg catgcatgga tcttggagga atcagatgtg cagtcaggga ccatagcaca
      121 agacaggcgt cttctactgg tgctaccagc aaatgctgga agccgggaac actgggtacg
      181 ttggaaacca cgtgatgtga agaagtaaga taaactgtag gagaaaagca tttcgtagtg
      241 ggccatgaag cctttcagga catgtattgc agtatgggcc ggcccattac gcaattggac
      301 gacaacaaag actagtatta gtaccacctc ggctatccac atagatcaaa gctgatttaa
      361 aagagttgtg cagatgatcc gtggcaggag accgaggtct cggttttaga gctagaaata
      421 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
      481 ttttgtttta gagctagaaa tagcaagtta aaataaggct agtccgtttt tagcgcgtgc
      541 atgcctgcag gtccccagat tagccttttc aatttcagaa agaatgctaa cccacagatg
      601 gttagagagg cttacgcagc agcactcatc aagacgatct acccgagcaa taatctccag
      661 gaaatcaaat accttcccaa gaaggttaaa gatgcagtca aaagattcag gactaactgc
      721 atcaagaaca cagagaaaga tatatttctc aagatcagaa gtactattcc agtatggacg
      781 attcaaggct tgcttcacaa accaaggcaa gtaatagaga ttggagtctc taaaaaggta
      841 gttcccactg aatcaaaggc catggagtca aagattcaaa tagaggacct aacagaactc
      901 gccgtaaaga ctggcgaaca gttcatacag agtctcttac gactcaatga caagaagaaa
      961 atcttcgtca acatggtgga gcacgacaca cttgtctact ccaaaaatat caaagataca
     1021 gtctcagaag accaaagggc aattgagact tttcaacaaa gggtaatatc cggaaacctc
     1081 ctcggattcc attgcccagc tatctgtcac tttattgtga agatagtgga aaaggaaggt
     1141 ggctcctaca aatgccatca ttgcgataaa ggaaaggcca tcgttgaaga tgcctctgcc
     1201 gacagtggtc ccaaagatgg acccccaccc acgaggagca tcgtggaaaa agaagacgtt
     1261 ccaaccacgt cttcaaagca agtggattga tgtgatatct ccactgacgt aagggatgac
     1321 gcacaatccc actatccttc gcaagaccct tcctctatat aaggaagttc atttcatttg
     1381 gagagaacac gggggactct agagttatca acaagtttgt acaaaaaagc aggcttcgaa
     1441 ggagatagaa ccaattctct aaggaaatac ttaaccatgg actataagga ccacgacgga
     1501 gactacaagg atcatgatat tgattacaaa gacgatgacg ataagatggc cccaaagaag
     1561 aagcggaagg tcggtatcca cggagtccca gcagccgaca agaagtacag catcggcctg
     1621 gacatcggca ccaactctgt gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc
     1681 aagaaattca aggtgctggg caacaccgac cggcacagca tcaagaagaa cctgatcgga
     1741 gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag aaccgccaga
     1801 agaagataca ccagacggaa gaaccggatc tgctatctgc aagagatctt cagcaacgag
     1861 atggccaagg tggacgacag cttcttccac agactggaag agtccttcct ggtggaagag
     1921 gataagaagc acgagcggca ccccatcttc ggcaacatcg tggacgaggt ggcctaccac
     1981 gagaagtacc ccaccatcta ccacctgaga aagaaactgg tggacagcac cgacaaggcc
     2041 gacctgcggc tgatctatct ggccctggcc cacatgatca agttccgggg ccacttcctg
     2101 atcgagggcg acctgaaccc cgacaacagc gacgtggaca agctgttcat ccagctggtg
     2161 cagacctaca accagctgtt cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag
     2221 gccatcctgt ctgccagact gagcaagagc agacggctgg aaaatctgat cgcccagctg
     2281 cccggcgaga agaagaatgg cctgttcgga aacctgattg ccctgagcct gggcctgacc
     2341 cccaacttca agagcaactt cgacctggcc gaggatgcca aactgcagct gagcaaggac
     2401 acctacgacg acgacctgga caacctgctg gcccagatcg gcgaccagta cgccgacctg
     2461 tttctggccg ccaagaacct gtccgacgcc atcctgctga gcgacatcct gagagtgaac
     2521 accgagatca ccaaggcccc cctgagcgcc tctatgatca agagatacga cgagcaccac
     2581 caggacctga ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa gtacaaagag
     2641 attttcttcg accagagcaa gaacggctac gccggctaca ttgacggcgg agccagccag
     2701 gaagagttct acaagttcat caagcccatc ctggaaaaga tggacggcac cgaggaactg
     2761 ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc ggaccttcga caacggcagc
     2821 atcccccacc agatccacct gggagagctg cacgccattc tgcggcggca ggaagatttt
     2881 tacccattcc tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc
     2941 tactacgtgg gccctctggc caggggaaac agcagattcg cctggatgac cagaaagagc
     3001 gaggaaacca tcaccccctg gaacttcgag gaagtggtgg acaagggcgc ttccgcccag
     3061 agcttcatcg agcggatgac caacttcgat aagaacctgc ccaacgagaa ggtgctgccc
     3121 aagcacagcc tgctgtacga gtacttcacc gtgtataacg agctgaccaa agtgaaatac
     3181 gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa ggccatcgtg
     3241 gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga ggactacttc
     3301 aagaaaatcg agtgcttcga ctccgtggaa atctccggcg tggaagatcg gttcaacgcc
     3361 tccctgggca cataccacga tctgctgaaa attatcaagg acaaggactt cctggacaat
     3421 gaggaaaacg aggacattct ggaagatatc gtgctgaccc tgacactgtt tgaggacaga
     3481 gagatgatcg aggaacggct gaaaacctat gcccacctgt tcgacgacaa agtgatgaag
     3541 cagctgaagc ggcggagata caccggctgg ggcaggctga gccggaagct gatcaacggc
     3601 atccgggaca agcagtccgg caagacaatc ctggatttcc tgaagtccga cggcttcgcc
     3661 aacagaaact tcatgcagct gatccacgac gacagcctga cctttaaaga ggacatccag
     3721 aaagcccagg tgtccggcca gggcgatagc ctgcacgagc acattgccaa tctggccggc
     3781 agccccgcca ttaagaaggg catcctgcag acagtgaagg tggtggacga gctcgtgaaa
     3841 gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga gaaccagacc
     3901 acccagaagg gacagaagaa cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa
     3961 gagctgggca gccagatcct gaaagaacac cccgtggaaa acacccagct gcagaacgag
     4021 aagctgtacc tgtactacct gcagaatggg cgggatatgt acgtggacca ggaactggac
     4081 atcaaccggc tgtccgacta cgatgtggac catatcgtgc ctcagagctt tctgaaggac
     4141 gactccatcg acaacaaggt gctgaccaga agcgacaaga accggggcaa gagcgacaac
     4201 gtgccctccg aagaggtcgt gaagaagatg aagaactact ggcggcagct gctgaacgcc
     4261 aagctgatta cccagagaaa gttcgacaat ctgaccaagg ccgagagagg cggcctgagc
     4321 gaactggata aggccggctt catcaagaga cagctggtgg aaacccggca gatcacaaag
     4381 cacgtggcac agatcctgga ctcccggatg aacactaagt acgacgagaa tgacaagctg
     4441 atccgggaag tgaaagtgat caccctgaag tccaagctgg tgtccgattt ccggaaggat
     4501 ttccagtttt acaaagtgcg cgagatcaac aactaccacc acgcccacga cgcctacctg
     4561 aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctggaaag cgagttcgtg
     4621 tacggcgact acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc
     4681 ggcaaggcta ccgccaagta cttcttctac agcaacatca tgaacttttt caagaccgag
     4741 attaccctgg ccaacggcga gatccggaag cggcctctga tcgagacaaa cggcgaaacc
     4801 ggggagatcg tgtgggataa gggccgggat tttgccaccg tgcggaaagt gctgagcatg
     4861 ccccaagtga atatcgtgaa aaagaccgag gtgcagacag gcggcttcag caaagagtct
     4921 atcctgccca agaggaacag cgataagctg atcgccagaa agaaggactg ggaccctaag
     4981 aagtacggcg gcttcgacag ccccaccgtg gcctattctg tgctggtggt ggccaaagtg
     5041 gaaaagggca agtccaagaa actgaagagt gtgaaagagc tgctggggat caccatcatg
     5101 gaaagaagca gcttcgagaa gaatcccatc gactttctgg aagccaaggg ctacaaagaa
     5161 gtgaaaaagg acctgatcat caagctgcct aagtactccc tgttcgagct ggaaaacggc
     5221 cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact ggccctgccc
     5281 tccaaatatg tgaacttcct gtacctggcc agccactatg agaagctgaa gggctccccc
     5341 gaggataatg agcagaaaca gctgtttgtg gaacagcaca agcactacct ggacgagatc
     5401 atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa tctggacaaa
     5461 gtgctgtccg cctacaacaa gcaccgggat aagcccatca gagagcaggc cgagaatatc
     5521 atccacctgt ttaccctgac caatctggga gcccctgccg ccttcaagta ctttgacacc
     5581 accatcgacc ggaagaggta caccagcacc aaagaggtgc tggacgccac cctgatccac
     5641 cagagcatca ccggcctgta cgagacacgg atcgacctgt ctcagctggg aggcgacaaa
     5701 aggccggcgg ccacgaaaaa ggccggccag gcaaaaaaga aaaagtaaga attcgcggcc
     5761 gcactcgaga tatctagacc cagctttctt gtacaaagtg gttgataaca gcgactacaa
     5821 ggatgacgat gacaaggctt agagctcgaa tttccccgat cgttcaaaca tttggcaata
     5881 aagtttctta agattgaatc ctgttgccgg tcttgcgatg attatcatat aatttctgtt
     5941 gaattacgtt aagcatgtaa taattaacat gtaatgcatg acgttattta tgagatgggt
     6001 ttttatgatt agagtcccgc aattatacat ttaatacgcg atagaaaaca aaatatagcg
     6061 cgcaaactag gataaattat cgcgcgcggt gtcatctatg ttactagatc gggaattcac
     6121 tggccgtcgt tttacactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     6181 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     6241 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gctagagcag
     6301 cttgagcttg gatcagattg tcgtttcccg ccttcagttt aaactatcag tgtttgacag
     6361 gatatattgg cgggtaaacc taagagaaaa gagcgtttat tagaataacg gatatttaaa
     6421 agggcgtgaa aaggtttatc cgttcgtcca tttgtatgtg catgccaacc acagggttcc
     6481 cctcgggatc aaagtacttt gatccaaccc ctccgctgct atagtgcagt cggcttctga
     6541 cgttcagtgc agccgtcttc tgaaaacgac atgtcgcaca agtcctaagt tacgcgacag
     6601 gctgccgccc tgcccttttc ctggcgtttt cttgtcgcgt gttttagtcg cataaagtag
     6661 aatacttgcg actagaaccg gagacattac gccatgaaca agagcgccgc cgctggcctg
     6721 ctgggctatg cccgcgtcag caccgacgac caggacttga ccaaccaacg ggccgaactg
     6781 cacgcggccg gctgcaccaa gctgttttcc gagaagatca ccggcaccag gcgcgaccgc
     6841 ccggagctgg ccaggatgct tgaccaccta cgccctggcg acgttgtgac agtgaccagg
     6901 ctagaccgcc tggcccgcag cacccgcgac ctactggaca ttgccgagcg catccaggag
     6961 gccggcgcgg gcctgcgtag cctggcagag ccgtgggccg acaccaccac gccggccggc
     7021 cgcatggtgt tgaccgtgtt cgccggcatt gccgagttcg agcgttccct aatcatcgac
     7081 cgcacccgga gcgggcgcga ggccgccaag gcccgaggcg tgaagtttgg cccccgccct
     7141 accctcaccc cggcacagat cgcgcacgcc cgcgagctga tcgaccagga aggccgcacc
     7201 gtgaaagagg cggctgcact gcttggcgtg catcgctcga ccctgtaccg cgcacttgag
     7261 cgcagcgagg aagtgacgcc caccgaggcc aggcggcgcg gtgccttccg tgaggacgca
     7321 ttgaccgagg ccgacgccct ggcggccgcc gagaatgaac gccaagagga acaagcatga
     7381 aaccgcacca ggacggccag gacgaaccgt ttttcattac cgaagagatc gaggcggaga
     7441 tgatcgcggc cgggtacgtg ttcgagccgc ccgcgcacgt ctcaaccgtg cggctgcatg
     7501 aaatcctggc cggtttgtct gatgccaagc tggcggcctg gccggccagc ttggccgctg
     7561 aagaaaccga gcgccgccgt ctaaaaaggt gatgtgtatt tgagtaaaac agcttgcgtc
     7621 atgcggtcgc tgcgtatatg atgcgatgag taaataaaca aatacgcaag gggaacgcat
     7681 gaaggttatc gctgtactta accagaaagg cgggtcaggc aagacgacca tcgcaaccca
     7741 tctagcccgc gccctgcaac tcgccggggc cgatgttctg ttagtcgatt ccgatcccca
     7801 gggcagtgcc cgcgattggg cggccgtgcg ggaagatcaa ccgctaaccg ttgtcggcat
     7861 cgaccgcccg acgattgacc gcgacgtgaa ggccatcggc cggcgcgact tcgtagtgat
     7921 cgacggagcg ccccaggcgg cggacttggc tgtgtccgcg atcaaggcag ccgacttcgt
     7981 gctgattccg gtgcagccaa gcccttacga catatgggcc accgccgacc tggtggagct
     8041 ggttaagcag cgcattgagg tcacggatgg aaggctacaa gcggcctttg tcgtgtcgcg
     8101 ggcgatcaaa ggcacgcgca tcggcggtga ggttgccgag gcgctggccg ggtacgagct
     8161 gcccattctt gagtcccgta tcacgcagcg cgtgagctac ccaggcactg ccgccgccgg
     8221 cacaaccgtt cttgaatcag aacccgaggg cgacgctgcc cgcgaggtcc aggcgctggc
     8281 cgctgaaatt aaatcaaaac tcatttgagt taatgaggta aagagaaaat gagcaaaagc
     8341 acaaacacgc taagtgccgg ccgtccgagc gcacgcagca gcaaggctgc aacgttggcc
     8401 agcctggcag acacgccagc catgaagcgg gtcaactttc agttgccggc ggaggatcac
     8461 accaagctga agatgtacgc ggtacgccaa ggcaagacca ttaccgagct gctatctgaa
     8521 tacatcgcgc agctaccaga gtaaatgagc aaatgaataa atgagtagat gaattttagc
     8581 ggctaaagga ggcggcatgg aaaatcaaga acaaccaggc accgacgccg tggaatgccc
     8641 catgtgtgga ggaacgggcg gttggccagg cgtaagcggc tgggttgtct gccggccctg
     8701 caatggcact ggaaccccca agcccgagga atcggcgtga cggtcgcaaa ccatccggcc
     8761 cggtacaaat cggcgcggcg ctgggtgatg acctggtgga gaagttgaag gccgcgcagg
     8821 ccgcccagcg gcaacgcatc gaggcagaag cacgccccgg tgaatcgtgg caagcggccg
     8881 ctgatcgaat ccgcaaagaa tcccggcaac cgccggcagc cggtgcgccg tcgattagga
     8941 agccgcccaa gggcgacgag caaccagatt ttttcgttcc gatgctctat gacgtgggca
     9001 cccgcgatag tcgcagcatc atggacgtgg ccgttttccg tctgtcgaag cgtgaccgac
     9061 gagctggcga ggtgatccgc tacgagcttc cagacgggca cgtagaggtt tccgcagggc
     9121 cggccggcat ggccagtgtg tgggattacg acctggtact gatggcggtt tcccatctaa
     9181 ccgaatccat gaaccgatac cgggaaggga agggagacaa gcccggccgc gtgttccgtc
     9241 cacacgttgc ggacgtactc aagttctgcc ggcgagccga tggcggaaag cagaaagacg
     9301 acctggtaga aacctgcatt cggttaaaca ccacgcacgt tgccatgcag cgtacgaaga
     9361 aggccaagaa cggccgcctg gtgacggtat ccgagggtga agccttgatt agccgctaca
     9421 agatcgtaaa gagcgaaacc gggcggccgg agtacatcga gatcgagcta gctgattgga
     9481 tgtaccgcga gatcacagaa ggcaagaacc cggacgtgct gacggttcac cccgattact
     9541 ttttgatcga tcccggcatc ggccgttttc tctaccgcct ggcacgccgc gccgcaggca
     9601 aggcagaagc cagatggttg ttcaagacga tctacgaacg cagtggcagc gccggagagt
     9661 tcaagaagtt ctgtttcacc gtgcgcaagc tgatcgggtc aaatgacctg ccggagtacg
     9721 atttgaagga ggaggcgggg caggctggcc cgatcctagt catgcgctac cgcaacctga
     9781 tcgagggcga agcatccgcc ggttcctaat gtacggagca gatgctaggg caaattgccc
     9841 tagcagggga aaaaggtcga aaagcactct ttcctgtgga tagcacgtac attgggaacc
     9901 caaagccgta cattgggaac cggaacccgt acattgggaa cccaaagccg tacattggga
     9961 accggtcaca catgtaagtg actgatataa aagagaaaaa aggcgatttt tccgcctaaa
    10021 actctttaaa acttattaaa actcttaaaa cccgcctggc ctgtgcataa ctgtctggcc
    10081 agcgcacagc cgaagagctg caaaaagcgc ctacccttcg gtcgctgcgc tccctacgcc
    10141 ccgccgcttc gcgtcggcct atcgcggccg ctggccgctc aaaaatggct ggcctacggc
    10201 caggcaatct accagggcgc ggacaagccg cgccgtcgcc actcgaccgc cggcgcccac
    10261 atcaaggcac cctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
    10321 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
    10381 ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat
    10441 agcggagtgt atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc
    10501 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt
    10561 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
    10621 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
    10681 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
    10741 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
    10801 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
    10861 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
    10921 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
    10981 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
    11041 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
    11101 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
    11161 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
    11221 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
    11281 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
    11341 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
    11401 tgcattctag gtactaaaac aattcatcca gtaaaatata atattttatt ttctcccaat
    11461 caggcttgat ccccagtaag tcaaaaaata gctcgacata ctgttcttcc ccgatatcct
    11521 ccctgatcga ccggacgcag aaggcaatgt cataccactt gtccgccctg ccgcttctcc
    11581 caagatcaat aaagccactt actttgccat ctttcacaaa gatgttgctg tctcccaggt
    11641 cgccgtggga aaagacaagt tcctcttcgg gcttttccgt ctttaaaaaa tcatacagct
    11701 cgcgcggatc tttaaatgga gtgtcttctt cccagttttc gcaatccaca tcggccagat
    11761 cgttattcag taagtaatcc aattcggcta agcggctgtc taagctattc gtatagggac
    11821 aatccgatat gtcgatggag tgaaagagcc tgatgcactc cgcatacagc tcgataatct
    11881 tttcagggct ttgttcatct tcatactctt ccgagcaaag gacgccatcg gcctcactca
    11941 tgagcagatt gctccagcca tcatgccgtt caaagtgcag gacctttgga acaggcagct
    12001 ttccttccag ccatagcatc atgtcctttt cccgttccac atcataggtg gtccctttat
    12061 accggctgtc cgtcattttt aaatataggt tttcattttc tcccaccagc ttatatacct
    12121 tagcaggaga cattccttcc gtatctttta cgcagcggta tttttcgatc agttttttca
    12181 attccggtga tattctcatt ttagccattt attatttcct tcctcttttc tacagtattt
    12241 aaagataccc caagaagcta attataacaa gacgaactcc aattcactgt tccttgcatt
    12301 ctaaaacctt aaataccaga aaacagcttt ttcaaagttg ttttcaaagt tggcgtataa
    12361 catagtatcg acggagccga ttttgaaacc gcggtgatca caggcagcaa cgctctgtca
    12421 tcgttacaat caacatgcta ccctccgcga gatcatccgt gtttcaaacc cggcagctta
    12481 gttgccgttc ttccgaatag catcggtaac atgagcaaag tctgccgcct tacaacggct
    12541 ctcccgctga cgccgtcccg gactgatggg ctgcctgtat cgagtggtga ttttgtgccg
    12601 agctgccggt cggggagctg ttggctggct ggtggcagga tatattgtgg tgtaaacaaa
    12661 ttgacgctta gacaacttaa taacacattg cggacgtttt taatgtactg aattaacgcc
    12721 gaattaattc gggggatctg gattttagta ctggattttg gttttaggaa ttagaaattt
    12781 tattgataga agtattttac aaatacaaat acatactaag ggtttcttat atgctcaaca
    12841 catgagcgaa accctatagg aaccctaatt cccttatctg ggaactactc acacattatt
    12901 atggagaaac tcgagcttgt cgatcgacag atccggtcgg catctactct atttctttgc
    12961 cctcggacga gtgctggggc gtcggtttcc actatcggcg agtacttcta cacagccatc
    13021 ggtccagacg gccgcgcttc tgcgggcgat ttgtgtacgc ccgacagtcc cggctccgga
    13081 tcggacgatt gcgtcgcatc gaccctgcgc ccaagctgca tcatcgaaat tgccgtcaac
    13141 caagctctga tagagttggt caagaccaat gcggagcata tacgcccgga gtcgtggcga
    13201 tcctgcaagc tccggatgcc tccgctcgaa gtagcgcgtc tgctgctcca tacaagccaa
    13261 ccacggcctc cagaagaaga tgttggcgac ctcgtattgg gaatccccga acatcgcctc
    13321 gctccagtca atgaccgctg ttatgcggcc attgtccgtc aggacattgt tggagccgaa
    13381 atccgcgtgc acgaggtgcc ggacttcggg gcagtcctcg gcccaaagca tcagctcatc
    13441 gagagcctgc gcgacggacg cactgacggt gtcgtccatc acagtttgcc agtgatacac
    13501 atggggatca gcaatcgcgc atatgaaatc acgccatgta gtgtattgac cgattccttg
    13561 cggtccgaat gggccgaacc cgctcgtctg gctaagatcg gccgcagcga tcgcatccat
    13621 agcctccgcg accggttgta gaacagcggg cagttcggtt tcaggcaggt cttgcaacgt
    13681 gacaccctgt gcacggcggg agatgcaata ggtcaggctc tcgctaaact ccccaatgtc
    13741 aagcacttcc ggaatcggga gcgcggccga tgcaaagtgc cgataaacat aacgatcttt
    13801 gtagaaacca tcggcgcagc tatttacccg caggacatat ccacgccctc ctacatcgaa
    13861 gctgaaagca cgagattctt cgccctccga gagctgcatc aggtcggaga cgctgtcgaa
    13921 cttttcgatc agaaacttct cgacagacgt cgcggtgagt tcaggctttt tcatatctca
    13981 ttgccccccg ggatctgcga aagctcgaga gagatagatt tgtagagaga gactggtgat
    14041 ttcagcgtgt cctctccaaa tgaaatgaac ttccttatat agaggaaggt cttgcgaagg
    14101 atagtgggat tgtgcgtcat cccttacgtc agtggagata tcacatcaat ccacttgctt
    14161 tgaagacgtg gttggaacgt cttctttttc cacgatgctc ctcgtgggtg ggggtccatc
    14221 tttgggacca ctgtcggcag aggcatcttg aacgatagcc tttcctttat cgcaatgatg
    14281 gcatttgtag gtgccacctt ccttttctac tgtccttttg atgaagtgac agatagctgg
    14341 gcaatggaat ccgaggaggt ttcccgatat taccctttgt tgaaaagtct caatagccct
    14401 ttggtcttct gagactgtat ctttgatatt cttggagtag acgagagtgt cgtgctccac
    14461 catgttatca catcaatcca cttgctttga agacgtggtt ggaacgtctt ctttttccac
    14521 gatgctcctc gtgggtgggg gtccatcttt gggaccactg tcggcagagg catcttgaac
    14581 gatagccttt cctttatcgc aatgatggca tttgtaggtg ccaccttcct tttctactgt
    14641 ccttttgatg aagtgacaga tagctgggca atggaatccg aggaggtttc ccgatattac
    14701 cctttgttga aaagtctcaa tagccctttg gtcttctgag actgtatctt tgatattctt
    14761 ggagtagacg agagtgtcgt gctccaccat gttggcaagc tgctctagcc aatacgcaaa
    14821 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
    14881 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
    14941 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
    15001 tttcacacag gaaacagcta tgaccatgat tacgcca
//