pTC223

LOCUS       pTC223               17961 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  Cas9/sgRNA targeting ANT1 locus, donor molecule with 5' homology 
            arm-Pnos:NptII-35S:ANT13' homology arm as a Gemini Viral Replicon 
            based on Bean Yellow Dwarf Virus; sgRNA= 7.
ACCESSION   .
VERSION     .
KEYWORDS    pTC223
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17961)
  AUTHORS   Cermak T, Baltes NJ, Cegan R, Zhang Y, Voytas DF
  TITLE     High-frequency, precise modification of the tomato genome.
  JOURNAL   Genome Biol. 2015 Nov 6;16(1):232. doi: 10.1186/s13059-015-0796-9.
  PUBMED    26541286
REFERENCE   2  (bases 1 to 17961)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..17961
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    1..25
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        966..1311
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             1345..5448
                     /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             5461..5481
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_RNA        6228..6303
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        7420..7599
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        7452..7635
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             7693..8490
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLL
                     SSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEH
                     QGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDI
                     ALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7750..7769)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8360..8379
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    8516..8690
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     promoter        8901..9246
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_feature    11684..11708
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             13008..13637
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             14071..15138
                     /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      15204..15398
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(15634..15656)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    15742..15882
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     15797..15816
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(15897..15914)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(16068..16656)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(16148..16167)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(16743..17537)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
ORIGIN
        1 tggcaggata tattgtggtg taaacaaatt gacgcttaga caacttaata acacattgcg
       61 gacgttttta atgtagagct caaagtttaa cgcgttagca gaaggcatgt tgttgtgact
      121 ccgaggggtt gcctcaaact ctatcttata accggcgtgg aggcatggag gcaggggtat
      181 tttggtcatt ttaatagata gtggaaaatg acgtggaatt tacttaaaga cgaagtcttt
      241 gcgacaaggg ggggcccacg ccgaatttaa tattaccggc gtggcccccc cttatcgcga
      301 gtgctttagc acgagcggtc cagatttaaa gtagaaaatt tcccgcccac tagggttaaa
      361 ggtgttcaca ctataaaagc atatacgatg tgatggtatt tgatggagcg tatattgtat
      421 caggtatttc cgttggatac gaattattcg tacgaccctc ggtaccgatc ggcgcgccag
      481 atttgccttt tcaatttcag aaagaatgct aacccacaga tggttagaga ggcttacgca
      541 gcaggtatca tcaagacgat ctacccgagc aataatctcc aggaaatcaa ataccttccc
      601 aagaaggtta aagatgcagt caaaagattc aggactaact gcatcaagaa cacagagaaa
      661 gatatatttc tcaagatcag aagtactatt ccagtatgga cgattcaagg cttgcttcac
      721 aaaccaaggc aagtaataga gattggagtc tctaaaaagg tagttcccac tgaatcaaag
      781 gccatggagt caaagattca aatagaggac ctaacagaac tcgccgtaaa gactggcgaa
      841 cagttcatac agagtctctt acgactcaat gacaagaaga aaatcttcgt caacatggtg
      901 gagcacgaca cacttgtcta ctccaaaaat atcaaagata cagtctcaga agaccaaagg
      961 gcaattgaga cttttcaaca aagggtaata tccggaaacc tcctcggatt ccattgccca
     1021 gctatctgtc actttattgt gaagatagtg gaaaaggaag gtggctccta caaatgccat
     1081 cattgcgata aaggaaaggc catcgttgaa gatgcctctg ccgacagtgg tcccaaagat
     1141 ggacccccac ccacgaggag catcgtggaa aaagaagacg ttccaaccac gtcttcaaag
     1201 caagtggatt gatgtgatat ctccactgac gtaagggatg acgcacaatc ccactatcct
     1261 tcgcaagacc cttcctctat ataaggaagt tcatttcatt tggagagaac acgggggact
     1321 cctgcaggta gatcgctcgt cgacatggat aagaagtact ctatcggact cgatatcgga
     1381 actaactctg tgggatgggc tgtgatcacc gatgagtaca aggtgccatc taagaagttc
     1441 aaggttctcg gaaacaccga taggcactct atcaagaaaa accttatcgg tgctctcctc
     1501 ttcgattctg gtgaaactgc tgaggctacc agactcaaga gaaccgctag aagaaggtac
     1561 accagaagaa agaacaggat ctgctacctc caagagatct tctctaacga gatggctaaa
     1621 gtggatgatt cattcttcca caggctcgaa gagtcattcc tcgtggaaga agataagaag
     1681 cacgagaggc accctatctt cggaaacatc gttgatgagg tggcatacca cgagaagtac
     1741 cctactatct accacctcag aaagaagctc gttgattcta ctgataaggc tgatctcagg
     1801 ctcatctacc tcgctctcgc tcacatgatc aagttcagag gacacttcct catcgagggt
     1861 gatctcaacc ctgataactc tgatgtggat aagttgttca tccagctcgt gcagacctac
     1921 aaccagcttt tcgaagagaa ccctatcaac gcttcaggtg tggatgctaa ggctatcctc
     1981 tctgctaggc tctctaagtc aagaaggctt gagaacctca ttgctcagct ccctggtgag
     2041 aagaagaacg gacttttcgg aaacttgatc gctctctctc tcggactcac ccctaacttc
     2101 aagtctaact tcgatctcgc tgaggatgca aagctccagc tctcaaagga tacctacgat
     2161 gatgatctcg ataacctcct cgctcagatc ggagatcagt acgctgattt gttcctcgct
     2221 gctaagaacc tctctgatgc tatcctcctc agtgatatcc tcagagtgaa caccgagatc
     2281 accaaggctc cactctcagc ttctatgatc aagagatacg atgagcacca ccaggatctc
     2341 acacttctca aggctcttgt tagacagcag ctcccagaga agtacaaaga gattttcttc
     2401 gatcagtcta agaacggata cgctggttac atcgatggtg gtgcatctca agaagagttc
     2461 tacaagttca tcaagcctat cctcgagaag atggatggaa ccgaggaact cctcgtgaag
     2521 ctcaatagag aggatcttct cagaaagcag aggaccttcg ataacggatc tatccctcat
     2581 cagatccacc tcggagagtt gcacgctatc cttagaaggc aagaggattt ctacccattc
     2641 ctcaaggata acagggaaaa gattgagaag attctcacct tcagaatccc ttactacgtg
     2701 ggacctctcg ctagaggaaa ctcaagattc gcttggatga ccagaaagtc tgaggaaacc
     2761 atcacccctt ggaacttcga agaggtggtg gataagggtg ctagtgctca gtctttcatc
     2821 gagaggatga ccaacttcga taagaacctt ccaaacgaga aggtgctccc taagcactct
     2881 ttgctctacg agtacttcac cgtgtacaac gagttgacca aggttaagta cgtgaccgag
     2941 ggaatgagga agcctgcttt tttgtcaggt gagcaaaaga aggctatcgt tgatctcttg
     3001 ttcaagacca acagaaaggt gaccgtgaag cagctcaaag aggattactt caagaaaatc
     3061 gagtgcttcg attcagttga gatttctggt gttgaggata ggttcaacgc atctctcgga
     3121 acctaccacg atctcctcaa gatcattaag gataaggatt tcttggataa cgaggaaaac
     3181 gaggatatct tggaggatat cgttcttacc ctcaccctct ttgaagatag agagatgatt
     3241 gaagaaaggc tcaagaccta cgctcatctc ttcgatgata aggtgatgaa gcagttgaag
     3301 agaagaagat acactggttg gggaaggctc tcaagaaagc tcattaacgg aatcagggat
     3361 aagcagtctg gaaagacaat ccttgatttc ctcaagtctg atggattcgc taacagaaac
     3421 ttcatgcagc tcatccacga tgattctctc acctttaaag aggatatcca gaaggctcag
     3481 gtttcaggac agggtgatag tctccatgag catatcgcta acctcgctgg atctcctgca
     3541 atcaagaagg gaatcctcca gactgtgaag gttgtggatg agttggtgaa ggtgatggga
     3601 aggcataagc ctgagaacat cgtgatcgaa atggctagag agaaccagac cactcagaag
     3661 ggacagaaga actctaggga aaggatgaag aggatcgagg aaggtatcaa agagcttgga
     3721 tctcagatcc tcaaagagca ccctgttgag aacactcagc tccagaatga gaagctctac
     3781 ctctactacc tccagaacgg aagggatatg tatgtggatc aagagttgga tatcaacagg
     3841 ctctctgatt acgatgttga tcatatcgtg ccacagtcat tcttgaagga tgattctatc
     3901 gataacaagg tgctcaccag gtctgataag aacaggggta agagtgataa cgtgccaagt
     3961 gaagaggttg tgaagaaaat gaagaactat tggaggcagc tcctcaacgc taagctcatc
     4021 actcagagaa agttcgataa cttgactaag gctgagaggg gaggactctc tgaattggat
     4081 aaggcaggat tcatcaagag gcagcttgtg gaaaccaggc agatcactaa gcacgttgca
     4141 cagatcctcg attctaggat gaacaccaag tacgatgaga acgataagtt gatcagggaa
     4201 gtgaaggtta tcaccctcaa gtcaaagctc gtgtctgatt tcagaaagga tttccaattc
     4261 tacaaggtga gggaaatcaa caactaccac cacgctcacg atgcttacct taacgctgtt
     4321 gttggaaccg ctctcatcaa gaagtatcct aagctcgagt cagagttcgt gtacggtgat
     4381 tacaaggtgt acgatgtgag gaagatgatc gctaagtctg agcaagagat cggaaaggct
     4441 accgctaagt atttcttcta ctctaacatc atgaatttct tcaagaccga gattaccctc
     4501 gctaacggtg agatcagaaa gaggccactc atcgagacaa acggtgaaac aggtgagatc
     4561 gtgtgggata agggaaggga tttcgctacc gttagaaagg tgctctctat gccacaggtg
     4621 aacatcgtta agaaaaccga ggtgcagacc ggtggattct ctaaagagtc tatcctccct
     4681 aagaggaact ctgataagct cattgctagg aagaaggatt gggaccctaa gaaatacggt
     4741 ggtttcgatt ctcctaccgt ggcttactct gttctcgttg tggctaaggt tgagaaggga
     4801 aagagtaaga agctcaagtc tgttaaggaa cttctcggaa tcactatcat ggaaaggtca
     4861 tctttcgaga agaacccaat cgatttcctc gaggctaagg gatacaaaga ggttaagaag
     4921 gatctcatca tcaagctccc aaagtactca ctcttcgaac tcgagaacgg tagaaagagg
     4981 atgctcgctt ctgctggtga gcttcaaaag ggaaacgagc ttgctctccc atctaagtac
     5041 gttaactttc tttacctcgc ttctcactac gagaagttga agggatctcc agaagataac
     5101 gagcagaagc aacttttcgt tgagcagcac aagcactact tggatgagat catcgagcag
     5161 atctctgagt tctctaaaag ggtgatcctc gctgatgcaa acctcgataa ggtgttgtct
     5221 gcttacaaca agcacagaga taagcctatc agggaacagg cagagaacat catccatctc
     5281 ttcaccctta ccaacctcgg tgctcctgct gctttcaagt acttcgatac aaccatcgat
     5341 aggaagagat acacctctac caaagaagtg ctcgatgcta ccctcatcca tcagtctatc
     5401 actggactct acgagactag gatcgatctc tcacagctcg gtggtgattc aagggctgat
     5461 cctaagaaga agaggaaggt ttgactcgag atatgaagat gaagatgaaa tatttggtgt
     5521 gtcaaataaa aagcttgtgt gcttaagttt gtgttttttt cttggcttgt tgtgttatga
     5581 atttgtggct ttttctaata ttaaatgaat gtaagatcac attataatga ataaacaaat
     5641 gtttctataa tccattgtga atgttttgtt ggatctcttc tgcagcatat aactactgta
     5701 tgtgctatgg tatggactat ggaatatgat taaagataag gagctccggt gacggaccca
     5761 tggcttcgtt gaacaacgga aactcgactt gccttccgca caatacatca tttcttctta
     5821 gctttttttc ttcttcttcg ttcatacagt ttttttttgt ttatcagctt acattttctt
     5881 gaaccgtagc tttcgttttc ttctttttaa ctttccattc ggagtttttg tatcttgttt
     5941 catagtttgt cccaggatta gaatgattag gcatcgaacc ttcaagaatt tgattgaata
     6001 aaacatcttc attcttaaga tatgaagata atcttcaaaa ggcccctggg aatctgaaag
     6061 aagagaagca ggcccattta tatgggaaag aacaatagta tttcttatat aggcccattt
     6121 aagttgaaaa caatcttcaa aagtcccaca tcgcttagat aagaaaacga agctgagttt
     6181 atatacagct agagtcgaag tagtgattgt cgtttataat ttgtagagtt ttagagctag
     6241 aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg
     6301 tgcttttttt cccggcgagc tttgggtacg tcacgtggct cgagcgcgta gtcctcggta
     6361 accacaacac ttgtcggtga gattatttaa tgctgattag attagacaaa aattaattag
     6421 ttttgagtag tggcgtaagt gtaaataatt agtctctttt ttaacttaga aaatagttta
     6481 atccttagta taaatagtca aaatcactgg aatgaaaaac agtttttaat ttttccaaat
     6541 ttgattctga taccatgtta aattcgtggt tcaaaatcac tgcaatgaaa agagcaatat
     6601 tgtttaactt tttttaggaa aatcgaattg atttatagtc agttgatata gagtgaatac
     6661 ataaggaaca tatacagttg atacaattgt ataattcgtt catacactta atacaaagtg
     6721 aacccacaag gaacatatac acttaatata attgtattcc ttgatacaaa ccaattttgt
     6781 tcgtgtctct actctctatt tcaatttcgc ttgactcttt actttttcta atatgtagct
     6841 ataaatcgta attaaacaat actatatctc taaatctctt attaagctca aactatggtc
     6901 atattcgaaa aaatcctttt aaatattggt cccctctcac gattaatgat agttataact
     6961 aacattcaaa ttttagttgt acttgacatc taaaacttaa aaaatagtac aagttaactt
     7021 tttctttttt ttaaaaaaag gaaatacttg tatttatttt tttaatatat agttatattt
     7081 ttggttattt gaaaatactt gatctgtcat gtatgctcag ttaaatatcg tcacattata
     7141 gagaaaaaag taataggaga aaaaaattaa aaattatttc gaaaaatcaa aatttttttt
     7201 tgattgaaat gaaagatggg tttcccaatc gaggctggca ggataggtac attgggaaat
     7261 ttggatttgt gtgttgaaaa tgattgttca atttggcttt tataacattt gtcgtgaagg
     7321 cgggaaacga caatctgatc atgagcggag aattaaggga gtcacgttat gacccccgcc
     7381 gatgacgcgg gacaagccgt tttacgtttg gaactgacag aaccgcaacg ttgaaggagc
     7441 cactcagccg cgggtttctg gagtttaatg agctaagcac atacgtcaga aaccattatt
     7501 gcgcgttcaa aagtcgccta aggtcactat cagctagcaa atatttcttg tcaaaaatgc
     7561 tccactgacg ttccataaat tcccctcggt atccaattag agtctcatat tcactctcaa
     7621 tccaaataat ctgcaccgga tccgctagag gatctcgacc tgcaagatcc cggggggcaa
     7681 tgagatccta ggatggggat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     7741 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     7801 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     7861 gccctgaatg aactccagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     7921 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     7981 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     8041 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     8101 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     8161 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     8221 gcgcgcatgc ccgacggcga ggatctcgtc gtgacacatg gcgatgcctg cttgccgaat
     8281 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     8341 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     8401 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     8461 ttctatcgcc ttcttgacga gttcttctga ttaattaacg atcgacaagc tctattttct
     8521 ccataataat gtgtgagtag ttcccagata agggaattag ggttcctata gggtttcgct
     8581 catgtgttga gcatataaga aacccttagt atgtatttgt atttgtaaaa tacttctatc
     8641 aataaaattt ctaattccta aaaccaaaat ccagtactaa aatccagatc ccccgaatta
     8701 attcggcgtt aattcagcat ggagtcaaag attcaaatag aggacctaac agaactcgcc
     8761 gtaaagactg gcgaacagtt catacagagt ctcttacgac tcaatgacaa gaagaaaatc
     8821 ttcgtcaaca tggtggagca cgacacactt gtctactcca aaaatatcaa agatacagtc
     8881 tcagaagacc aaagggcaat tgagactttt caacaaaggg taatatccgg aaacctcctc
     8941 ggattccatt gcccagctat ctgtcacttt attgtgaaga tagtggaaaa ggaaggtggc
     9001 tcctacaaat gccatcattg cgataaagga aaggccatcg ttgaagatgc ctctgccgac
     9061 agtggtccca aagatggacc cccacccacg aggagcatcg tggaaaaaga agacgttcca
     9121 accacgtctt caaagcaagt ggattgatgt gatatctcca ctgacgtaag ggatgacgca
     9181 caatcccact atccttcgca agacccttcc tctatataag gaagttcatt tcatttggag
     9241 agaacacggg ggactcttga cagtagtata atatattatc aaattattat gaacagtaca
     9301 tctatgtctt cattgggagt gagaaaaggt tcatggactg atgaagaaga ttttcttcta
     9361 agaaaatgta ttgataagta tggtgaagga aaatggcatc ttgttcccat aagagctggt
     9421 aactattaaa ttaactatca cgttattttt atttgtcttt ctgtctcatt ttatttgacg
     9481 ttattacgaa tatcatctga aaatgtacgt gcaggtctga atagatgtcg gaaaagttgt
     9541 agattgaggt ggctgaatta tctaaggcca catatcaaga gaggtgactt tgaacaagat
     9601 gaagtggatc tcattttgag gcttcataag ctcttaggca acaggcatgc aagtttatgt
     9661 tttgacaaaa tttgattagt atatattata tatacgtgtg actatttcat ctaaatgtta
     9721 cgttatttta cgtagatggt cacttattgc tggtagactt cccggaagga cagctaacga
     9781 tgtgaaaaac tattggaaca ctaatcttct aaggaagtta aatactacta aaattgttcc
     9841 tcgcgaaaag attaacaata agtgtggaga aattagtact aagattgaaa ttataaaacc
     9901 tcaacgacgc aagtatttct caagcacaat gaagaatgtt acaaacaata atgtaatttt
     9961 ggacgaggag gaacattgca tttaaatgag tgtacttcaa gtcacaccgg cgagtgtttg
    10021 atcgccggcg gtaccgagtg tacttcaagt cagtgggaaa tcaataaaat gattatttta
    10081 tgaatatatt tcattgtgca agtagataga aattacatat gttacataac acacgaaata
    10141 aacaaaaaaa gacaatccaa aaacaaacac cccaaaaaaa ataatcactt tagataaact
    10201 cgtatgagga gaggcacgtt cagtgactcg acgattcccg agcaaaaaaa gtctccccgt
    10261 cacacatgta gtgggtgacg caattatctt taaagtaatc cttctgttga cttgtcattg
    10321 ataacatcca gtcttcgtca ggattgcaaa gaattataga agggatccca ccttttattt
    10381 tcttcttttt tccatattta gggttgacag tgaaatcaga ctggcaacct attaattgct
    10441 tccacaatgg gacgaacttg aaggggatgt cgtcgatgat attataggtg gcgtgttcat
    10501 cgtagttggt gaaatcgatg gtaccgttcc aatagttgtg tcgtccgaga cttctagccc
    10561 aggtggtctt tccggtacga gttggtccgc agatgtagag gctggggtgt cggattccat
    10621 tccttccatt gtccttgtta aatcggccat ccattcaagg tcagattgag cttgttggta
    10681 tgagacagga tgtatgtaag tataagcgtc tatgcttaca tggtatagat gggtttccct
    10741 ccaggagtgt agatcttcgt ggcagcgaag atctgattct gtgaagggcg acacatacgg
    10801 ttcaggttgt ggagggaata atttgttggc tgaatattcc agccattgaa gctttgttgc
    10861 ccattcatga gggaattctt ccttgatcat gtcaagatat tcctccttag acgttgcagt
    10921 ctggataata gttctccatc gtgcgtcaga tttgcgagga gaaaccttat gatctcggaa
    10981 atctcctctg gttttaatat ctccgtcctt tgatatgtaa tcaaggactt gtttagagtt
    11041 tctagctggc tggatattag ggtgatttcc ttcaaaatcg aaaaaagaag gatccctaat
    11101 acaaggtttt ttatcaagct ggagaagagc atgatagtgg gtagtgccat cttgatgaag
    11161 ctcagaagca acaccaagga agaaaataag aaaaggtgtg agtttctccc agagaaactg
    11221 gaataaatca tctctttgag atgagcactt gggataggta aggaaaacat atttagattg
    11281 gagtctgaag ttcttactag cagaaggcat gttgttgtga ctccgagggg ttgcctcaaa
    11341 ctctatctta taaccggcgt ggaggcatgg aggcaggggt attttggtca ttttaataga
    11401 tagtggaaaa tgacgtggaa tttacttaaa gacgaagtct ttgcgacaag ggggggccca
    11461 cgccgaattt aatattaccg gcgtggcccc cccttatcgc gagtgcttta gcacgagcgg
    11521 tccagattta aagtagaaaa tttcccgccc actagggtta aaggtgttca cactataaaa
    11581 gcatatacga tgtgatggta tttgatggag cgtatattgt atcaggtatt tccgttggat
    11641 acgaattatt cgtacgaccc tcatagttta aactatcagt gtttgacagg atatattggc
    11701 gggtaaacct aagagaaaag agcgtttatt agaataacgg atatttaaaa gggcgtgaaa
    11761 aggtttatcc gttcgtccat ttgtatgtgc atgccaacca cagggttccc ctcgggatca
    11821 aagtactttg atccaacccc tccgctgcta tagtgcagtc ggcttctgac gttcagtgca
    11881 gccgtcttct gaaaacgaca tgtcgcacaa gtcctaagtt acgcgacagg ctgccgccct
    11941 gcccttttcc tggcgttttc ttgtcgcgtg ttttagtcgc ataaagtaga atacttgcga
    12001 ctagaaccgg agacattacg ccatgaacaa gagcgccgcc gctggcctgc tgggctatgc
    12061 ccgcgtcagc accgacgacc aggacttgac caaccaacgg gccgaactgc acgcggccgg
    12121 ctgcaccaag ctgttttccg agaagatcac cggcaccagg cgcgaccgcc cggagctggc
    12181 caggatgctt gaccacctac gccctggcga cgttgtgaca gtgaccaggc tagaccgcct
    12241 ggcccgcagc acccgcgacc tactggacat tgccgagcgc atccaggagg ccggcgcggg
    12301 cctgcgtagc ctggcagagc cgtgggccga caccaccacg ccggccggcc gcatggtgtt
    12361 gaccgtgttc gccggcattg ccgagttcga gcgttcccta atcatcgacc gcacccggag
    12421 cgggcgcgag gccgccaagg cccgaggcgt gaagtttggc ccccgcccta ccctcacccc
    12481 ggcacagatc gcgcacgccc gcgagctgat cgaccaggaa ggccgcaccg tgaaagaggc
    12541 ggctgcactg cttggcgtgc atcgctcgac cctgtaccgc gcacttgagc gcagcgagga
    12601 agtgacgccc accgaggcca ggcggcgcgg tgccttccgt gaggacgcat tgaccgaggc
    12661 cgacgccctg gcggccgccg agaatgaacg ccaagaggaa caagcatgaa accgcaccag
    12721 gacggccagg acgaaccgtt tttcattacc gaagagatcg aggcggagat gatcgcggcc
    12781 gggtacgtgt tcgagccgcc cgcgcacggc tcaaccgtgc ggctgcatga aatcctggcc
    12841 ggtttgtctg atgccaagct ggcggcctgg ccggccagct tggccgctga agaaaccgag
    12901 cgccgccgtc taaaaaggtg atgtgtattt gagtaaaaca gcttgcgtca tgcggtcgct
    12961 gcgtatatga tgcgatgagt aaataaacaa atacgcaagg ggaacgcatg aaggttatcg
    13021 ctgtacttaa ccagaaaggc gggtcaggca agacgaccat cgcaacccat ctagcccgcg
    13081 ccctgcaact cgccggggcc gatgttctgt tagtcgattc cgatccccag ggcagtgccc
    13141 gcgattgggc ggccgtgcgg gaagatcaac cgctaaccgt tgtcggcatc gaccgcccga
    13201 cgattgaccg cgacgtgaag gccatcggcc ggcgcgactt cgtagtgatc gacggagcgc
    13261 cccaggcggc ggacttggct gtgtccgcga tcaaggcagc cgacttcgtg ctgattccgg
    13321 tgcagccaag cccttacgac atatgggcca ccgccgacct ggtggagctg gttaagcagc
    13381 gcattgaggt cacggatgga aggctacaag cggcctttgt cgtgtcgcgg gcgatcaaag
    13441 gcacgcgcat cggcggtgag gttgccgagg cgctggccgg gtacgagctg cccattcttg
    13501 agtcccgtat cacgcagcgc gtgagctacc caggcactgc cgccgccggc acaaccgttc
    13561 ttgaatcaga acccgagggc gacgctgccc gcgaggtcca ggcgctggcc gctgaaatta
    13621 aatcaaaact catttgagtt aatgaggtaa agagaaaatg agcaaaagca caaacacgct
    13681 aagtgccggc cgtccgagcg cacgcagcag caaggctgca acgttggcca gcctggcaga
    13741 cacgccagcc atgaagcggg tcaactttca gttgccggcg gaggatcaca ccaagctgaa
    13801 gatgtacgcg gtacgccaag gcaagaccat taccgagctg ctatctgaat acatcgcgca
    13861 gctaccagag taaatgagca aatgaataaa tgagtagatg aattttagcg gctaaaggag
    13921 gcggcatgga aaatcaagaa caaccaggca ccgacgccgt ggaatgcccc atgtgtggag
    13981 gaacgggcgg ttggccaggc gtaagcggct gggttgtctg ccggccctgc aatggcactg
    14041 gaacccccaa gcccgaggaa tcggcgtgac ggtcgcaaac catccggccc ggtacaaatc
    14101 ggcgcggcgc tgggtgatga cctggtggag aagttgaagg ccgcgcaggc cgcccagcgg
    14161 caacgcatcg aggcagaagc acgccccggt gaatcgtggc aagcggccgc tgatcgaatc
    14221 cgcaaagaat cccggcaacc gccggcagcc ggtgcgccgt cgattaggaa gccgcccaag
    14281 ggcgacgagc aaccagattt tttcgttccg atgctctatg acgtgggcac ccgcgatagt
    14341 cgcagcatca tggacgtggc cgttttccgt ctgtcgaagc gtgaccgacg agctggcgag
    14401 gtgatccgct acgagcttcc agacgggcac gtagaggttt ccgcagggcc ggccggcatg
    14461 gccagtgtgt gggattacga cctggtactg atggcggttt cccatctaac cgaatccatg
    14521 aaccgatacc gggaagggaa gggagacaag cccggccgcg tgttccgtcc acacgttgcg
    14581 gacgtactca agttctgccg gcgagccgat ggcggaaagc agaaagacga cctggtagaa
    14641 acctgcattc ggttaaacac cacgcacgtt gccatgcagc gtacgaagaa ggccaagaac
    14701 ggccgcctgg tgacggtatc cgagggtgaa gccttgatta gccgctacaa gatcgtaaag
    14761 agcgaaaccg ggcggccgga gtacatcgag atcgagctag ctgattggat gtaccgcgag
    14821 atcacagaag gcaagaaccc ggacgtgctg acggttcacc ccgattactt tttgatcgat
    14881 cccggcatcg gccgttttct ctaccgcctg gcacgccgcg ccgcaggcaa ggcagaagcc
    14941 agatggttgt tcaagacgat ctacgaacgc agtggcagcg ccggagagtt caagaagttc
    15001 tgtttcaccg tgcgcaagct gatcgggtca aatgacctgc cggagtacga tttgaaggag
    15061 gaggcggggc aggctggccc gatcctagtc atgcgctacc gcaacctgat cgagggcgaa
    15121 gcatccgccg gttcctaatg tacggagcag atgctagggc aaattgccct agcaggggaa
    15181 aaaggtcgaa aaggcctctt tcctgtggat agcacgtaca ttgggaaccc aaagccgtac
    15241 attgggaacc ggaacccgta cattgggaac ccaaagccgt acattgggaa ccggtcacac
    15301 atgtaagtga ctgatataaa agagaaaaaa ggcgattttt ccgcctaaaa ctctttaaaa
    15361 cttattaaaa ctcttaaaac ccgcctggcc tgtgcataac tgtctggcca gcgcacagcc
    15421 gaagagctgc aaaaagcgcc tacccttcgg tcgctgcgct ccctacgccc cgccgcttcg
    15481 cgtcggccta tcgcggccgc tggccgctca aaaatggctg gcctacggcc aggcaatcta
    15541 ccagggcgcg gacaagccgc gccgtcgcca ctcgaccgcc ggcgcccaca tcaaggcacc
    15601 ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggaaac
    15661 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
    15721 gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta
    15781 tactggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
    15841 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg
    15901 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
    15961 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
    16021 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
    16081 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
    16141 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
    16201 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
    16261 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
    16321 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
    16381 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
    16441 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
    16501 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
    16561 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
    16621 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
    16681 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gcattctagg
    16741 tactaaaaca attcatccag taaaatataa tattttattt tctcccaatc aggcttgatc
    16801 cccagtaagt caaaaaatag ctcgacatac tgttcttccc cgatatcctc cctgatcgac
    16861 cggacgcaga aggcaatgtc ataccacttg tccgccctgc cgcttctccc aagatcaata
    16921 aagccactta ctttgccatc tttcacaaag atgttgctgt ctcccaggtc gccgtgggaa
    16981 aagacaagtt cctcttcggg cttttccgtc tttaaaaaat catacagctc gcgcggatct
    17041 ttaaatggag tgtcttcttc ccagttttcg caatccacat cggccagatc gttattcagt
    17101 aagtaatcca attcggctaa gcggctgtct aagctattcg tatagggaca atccgatatg
    17161 tcgatggagt gaaagagcct gatgcactcc gcatacagct cgataatctt ttcagggctt
    17221 tgttcatctt catactcttc cgagcaaagg acgccatcgg cctcactcat gagcagattg
    17281 ctccagccat catgccgttc aaagtgcagg acctttggaa caggcagctt tccttccagc
    17341 catagcatca tgtccttttc ccgttccaca tcataggtgg tccctttata ccggctgtcc
    17401 gtcattttta aatataggtt ttcattttct cccaccagct tatatacctt agcaggagac
    17461 attccttccg tatcttttac gcagcggtat ttttcgatca gttttttcaa ttccggtgat
    17521 attctcattt tagccattta ttatttcctt cctcttttct acagtattta aagatacccc
    17581 aagaagctaa ttataacaag acgaactcca attcactgtt ccttgcattc taaaacctta
    17641 aataccagaa aacagctttt tcaaagttgt tttcaaagtt ggcgtataac atagtatcga
    17701 cggagccgat tttgaaaccg cggtgatcac aggcagcaac gctctgtcat cgttacaatc
    17761 aacatgctac cctccgcgag atcatccgtg tttcaaaccc ggcagcttag ttgccgttct
    17821 tccgaatagc atcggtaaca tgagcaaagt ctgccgcctt acaacggctc tcccgctgac
    17881 gccgtcccgg actgatgggc tgcctgtatc gagtggtgat tttgtgccga gctgccggtc
    17941 ggggagctgt tggctggctg g
//