pTC217

LOCUS       pTC217              17997 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= 1b.
ACCESSION   .
VERSION     .
KEYWORDS    pTC217
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17997)
  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 17997)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..17997
                     /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        6229..6304
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        7456..7635
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        7488..7671
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             7729..8526
                     /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(7786..7805)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8396..8415
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    8552..8726
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     promoter        8937..9282
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_feature    11720..11744
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             13044..13673
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             14107..15174
                     /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      15240..15434
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(15670..15692)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    15778..15918
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     15833..15852
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(15933..15950)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(16104..16692)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(16184..16203)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(16779..17573)
                     /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 tagtgattga caatttaata caccttttgt tttagagcta
     6241 gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg
     6301 gtgctttttt tcccggcgag ctttgggtac gtcacgtggc tcgagcgcgt agtcctcggt
     6361 aaccacaaca cttgtcggtg agattattta atgctgatta gattagacaa aaattaatta
     6421 gttttgagta gtggcgtaag tgtaaataat tagtctcttt tttaacttag aaaatagttt
     6481 aatccttagt ataaatagtc aaaatcactg gaatgaaaaa cagtttttaa tttttccaaa
     6541 tttgattctg ataccatgtt aaattcgtgg ttcaaaatca ctgcaatgaa aagagcaata
     6601 ttgtttaact ttttttagga aaatcgaatt gatttatagt cagttgatat agagtgaata
     6661 cataaggaac atatacagtt gatacaattg tataattcgt tcatacactt aatacaaagt
     6721 gaacccacaa ggaacatata cacttaatat aattgtattc cttgatacaa accaattttg
     6781 ttcgtgtctc tactctctat ttcaatttcg cttgactctt tactttttct aatatgtagc
     6841 tataaatcgt aattaaacaa tactatatct ctaaatctct tattaagctc aaactatggt
     6901 catattcgaa aaaatccttt taaatattgg tcccctctca cgattaatga tagttataac
     6961 taacattcaa attttagttg tacttgacat ctaaaactta aaaaatagta caagttaact
     7021 ttttcttttt tttaaaaaaa ggaaatactt gtatttattt ttttaatata tagttatatt
     7081 tttggttatt tgaaaatact tgatctgtca tgtatgctca gttaaatatc gtcacattat
     7141 agagaaaaaa gtaataggag aaaaaaatta aaaattattt cgaaaaatca aaattttttt
     7201 ttgattgaaa tgaaagatgg gtttcccaat cgaggctggc aggataggta cattgggaaa
     7261 tttggatttg tgtgttgaaa atgattgttc aatttggctt ttataacatt tgtcgtttat
     7321 aatttgtaga aggctctcta caagttggac tgaaggcggg aaacgacaat ctgatcatga
     7381 gcggagaatt aagggagtca cgttatgacc cccgccgatg acgcgggaca agccgtttta
     7441 cgtttggaac tgacagaacc gcaacgttga aggagccact cagccgcggg tttctggagt
     7501 ttaatgagct aagcacatac gtcagaaacc attattgcgc gttcaaaagt cgcctaaggt
     7561 cactatcagc tagcaaatat ttcttgtcaa aaatgctcca ctgacgttcc ataaattccc
     7621 ctcggtatcc aattagagtc tcatattcac tctcaatcca aataatctgc accggatccg
     7681 ctagaggatc tcgacctgca agatcccggg gggcaatgag atcctaggat ggggattgaa
     7741 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     7801 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     7861 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact ccaggacgag
     7921 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     7981 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     8041 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     8101 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     8161 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     8221 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     8281 ctcgtcgtga cacatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     8341 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     8401 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     8461 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     8521 ttctgattaa ttaacgatcg acaagctcta ttttctccat aataatgtgt gagtagttcc
     8581 cagataaggg aattagggtt cctatagggt ttcgctcatg tgttgagcat ataagaaacc
     8641 cttagtatgt atttgtattt gtaaaatact tctatcaata aaatttctaa ttcctaaaac
     8701 caaaatccag tactaaaatc cagatccccc gaattaattc ggcgttaatt cagcatggag
     8761 tcaaagattc aaatagagga cctaacagaa ctcgccgtaa agactggcga acagttcata
     8821 cagagtctct tacgactcaa tgacaagaag aaaatcttcg tcaacatggt ggagcacgac
     8881 acacttgtct actccaaaaa tatcaaagat acagtctcag aagaccaaag ggcaattgag
     8941 acttttcaac aaagggtaat atccggaaac ctcctcggat tccattgccc agctatctgt
     9001 cactttattg tgaagatagt ggaaaaggaa ggtggctcct acaaatgcca tcattgcgat
     9061 aaaggaaagg ccatcgttga agatgcctct gccgacagtg gtcccaaaga tggaccccca
     9121 cccacgagga gcatcgtgga aaaagaagac gttccaacca cgtcttcaaa gcaagtggat
     9181 tgatgtgata tctccactga cgtaagggat gacgcacaat cccactatcc ttcgcaagac
     9241 ccttcctcta tataaggaag ttcatttcat ttggagagaa cacgggggac tcttgacagt
     9301 agtataatat attatcaaat tattatgaac agtacatcta tgtcttcatt gggagtgaga
     9361 aaaggttcat ggactgatga agaagatttt cttctaagaa aatgtattga taagtatggt
     9421 gaaggaaaat ggcatcttgt tcccataaga gctggtaact attaaattaa ctatcacgtt
     9481 atttttattt gtctttctgt ctcattttat ttgacgttat tacgaatatc atctgaaaat
     9541 gtacgtgcag gtctgaatag atgtcggaaa agttgtagat tgaggtggct gaattatcta
     9601 aggccacata tcaagagagg tgactttgaa caagatgaag tggatctcat tttgaggctt
     9661 cataagctct taggcaacag gcatgcaagt ttatgttttg acaaaatttg attagtatat
     9721 attatatata cgtgtgacta tttcatctaa atgttacgtt attttacgta gatggtcact
     9781 tattgctggt agacttcccg gaaggacagc taacgatgtg aaaaactatt ggaacactaa
     9841 tcttctaagg aagttaaata ctactaaaat tgttcctcgc gaaaagatta acaataagtg
     9901 tggagaaatt agtactaaga ttgaaattat aaaacctcaa cgacgcaagt atttctcaag
     9961 cacaatgaag aatgttacaa acaataatgt aattttggac gaggaggaac attgcattta
    10021 aatgagtgta cttcaagtca caccggcgag tgtttgatcg ccggcggtac cgagtgtact
    10081 tcaagtcagt gggaaatcaa taaaatgatt attttatgaa tatatttcat tgtgcaagta
    10141 gatagaaatt acatatgtta cataacacac gaaataaaca aaaaaagaca atccaaaaac
    10201 aaacacccca aaaaaaataa tcactttaga taaactcgta tgaggagagg cacgttcagt
    10261 gactcgacga ttcccgagca aaaaaagtct ccccgtcaca catgtagtgg gtgacgcaat
    10321 tatctttaaa gtaatccttc tgttgacttg tcattgataa catccagtct tcgtcaggat
    10381 tgcaaagaat tatagaaggg atcccacctt ttattttctt cttttttcca tatttagggt
    10441 tgacagtgaa atcagactgg caacctatta attgcttcca caatgggacg aacttgaagg
    10501 ggatgtcgtc gatgatatta taggtggcgt gttcatcgta gttggtgaaa tcgatggtac
    10561 cgttccaata gttgtgtcgt ccgagacttc tagcccaggt ggtctttccg gtacgagttg
    10621 gtccgcagat gtagaggctg gggtgtcgga ttccattcct tccattgtcc ttgttaaatc
    10681 ggccatccat tcaaggtcag attgagcttg ttggtatgag acaggatgta tgtaagtata
    10741 agcgtctatg cttacatggt atagatgggt ttccctccag gagtgtagat cttcgtggca
    10801 gcgaagatct gattctgtga agggcgacac atacggttca ggttgtggag ggaataattt
    10861 gttggctgaa tattccagcc attgaagctt tgttgcccat tcatgaggga attcttcctt
    10921 gatcatgtca agatattcct ccttagacgt tgcagtctgg ataatagttc tccatcgtgc
    10981 gtcagatttg cgaggagaaa ccttatgatc tcggaaatct cctctggttt taatatctcc
    11041 gtcctttgat atgtaatcaa ggacttgttt agagtttcta gctggctgga tattagggtg
    11101 atttccttca aaatcgaaaa aagaaggatc cctaatacaa ggttttttat caagctggag
    11161 aagagcatga tagtgggtag tgccatcttg atgaagctca gaagcaacac caaggaagaa
    11221 aataagaaaa ggtgtgagtt tctcccagag aaactggaat aaatcatctc tttgagatga
    11281 gcacttggga taggtaagga aaacatattt agattggagt ctgaagttct tactagcaga
    11341 aggcatgttg ttgtgactcc gaggggttgc ctcaaactct atcttataac cggcgtggag
    11401 gcatggaggc aggggtattt tggtcatttt aatagatagt ggaaaatgac gtggaattta
    11461 cttaaagacg aagtctttgc gacaaggggg ggcccacgcc gaatttaata ttaccggcgt
    11521 ggccccccct tatcgcgagt gctttagcac gagcggtcca gatttaaagt agaaaatttc
    11581 ccgcccacta gggttaaagg tgttcacact ataaaagcat atacgatgtg atggtatttg
    11641 atggagcgta tattgtatca ggtatttccg ttggatacga attattcgta cgaccctcat
    11701 agtttaaact atcagtgttt gacaggatat attggcgggt aaacctaaga gaaaagagcg
    11761 tttattagaa taacggatat ttaaaagggc gtgaaaaggt ttatccgttc gtccatttgt
    11821 atgtgcatgc caaccacagg gttcccctcg ggatcaaagt actttgatcc aacccctccg
    11881 ctgctatagt gcagtcggct tctgacgttc agtgcagccg tcttctgaaa acgacatgtc
    11941 gcacaagtcc taagttacgc gacaggctgc cgccctgccc ttttcctggc gttttcttgt
    12001 cgcgtgtttt agtcgcataa agtagaatac ttgcgactag aaccggagac attacgccat
    12061 gaacaagagc gccgccgctg gcctgctggg ctatgcccgc gtcagcaccg acgaccagga
    12121 cttgaccaac caacgggccg aactgcacgc ggccggctgc accaagctgt tttccgagaa
    12181 gatcaccggc accaggcgcg accgcccgga gctggccagg atgcttgacc acctacgccc
    12241 tggcgacgtt gtgacagtga ccaggctaga ccgcctggcc cgcagcaccc gcgacctact
    12301 ggacattgcc gagcgcatcc aggaggccgg cgcgggcctg cgtagcctgg cagagccgtg
    12361 ggccgacacc accacgccgg ccggccgcat ggtgttgacc gtgttcgccg gcattgccga
    12421 gttcgagcgt tccctaatca tcgaccgcac ccggagcggg cgcgaggccg ccaaggcccg
    12481 aggcgtgaag tttggccccc gccctaccct caccccggca cagatcgcgc acgcccgcga
    12541 gctgatcgac caggaaggcc gcaccgtgaa agaggcggct gcactgcttg gcgtgcatcg
    12601 ctcgaccctg taccgcgcac ttgagcgcag cgaggaagtg acgcccaccg aggccaggcg
    12661 gcgcggtgcc ttccgtgagg acgcattgac cgaggccgac gccctggcgg ccgccgagaa
    12721 tgaacgccaa gaggaacaag catgaaaccg caccaggacg gccaggacga accgtttttc
    12781 attaccgaag agatcgaggc ggagatgatc gcggccgggt acgtgttcga gccgcccgcg
    12841 cacggctcaa ccgtgcggct gcatgaaatc ctggccggtt tgtctgatgc caagctggcg
    12901 gcctggccgg ccagcttggc cgctgaagaa accgagcgcc gccgtctaaa aaggtgatgt
    12961 gtatttgagt aaaacagctt gcgtcatgcg gtcgctgcgt atatgatgcg atgagtaaat
    13021 aaacaaatac gcaaggggaa cgcatgaagg ttatcgctgt acttaaccag aaaggcgggt
    13081 caggcaagac gaccatcgca acccatctag cccgcgccct gcaactcgcc ggggccgatg
    13141 ttctgttagt cgattccgat ccccagggca gtgcccgcga ttgggcggcc gtgcgggaag
    13201 atcaaccgct aaccgttgtc ggcatcgacc gcccgacgat tgaccgcgac gtgaaggcca
    13261 tcggccggcg cgacttcgta gtgatcgacg gagcgcccca ggcggcggac ttggctgtgt
    13321 ccgcgatcaa ggcagccgac ttcgtgctga ttccggtgca gccaagccct tacgacatat
    13381 gggccaccgc cgacctggtg gagctggtta agcagcgcat tgaggtcacg gatggaaggc
    13441 tacaagcggc ctttgtcgtg tcgcgggcga tcaaaggcac gcgcatcggc ggtgaggttg
    13501 ccgaggcgct ggccgggtac gagctgccca ttcttgagtc ccgtatcacg cagcgcgtga
    13561 gctacccagg cactgccgcc gccggcacaa ccgttcttga atcagaaccc gagggcgacg
    13621 ctgcccgcga ggtccaggcg ctggccgctg aaattaaatc aaaactcatt tgagttaatg
    13681 aggtaaagag aaaatgagca aaagcacaaa cacgctaagt gccggccgtc cgagcgcacg
    13741 cagcagcaag gctgcaacgt tggccagcct ggcagacacg ccagccatga agcgggtcaa
    13801 ctttcagttg ccggcggagg atcacaccaa gctgaagatg tacgcggtac gccaaggcaa
    13861 gaccattacc gagctgctat ctgaatacat cgcgcagcta ccagagtaaa tgagcaaatg
    13921 aataaatgag tagatgaatt ttagcggcta aaggaggcgg catggaaaat caagaacaac
    13981 caggcaccga cgccgtggaa tgccccatgt gtggaggaac gggcggttgg ccaggcgtaa
    14041 gcggctgggt tgtctgccgg ccctgcaatg gcactggaac ccccaagccc gaggaatcgg
    14101 cgtgacggtc gcaaaccatc cggcccggta caaatcggcg cggcgctggg tgatgacctg
    14161 gtggagaagt tgaaggccgc gcaggccgcc cagcggcaac gcatcgaggc agaagcacgc
    14221 cccggtgaat cgtggcaagc ggccgctgat cgaatccgca aagaatcccg gcaaccgccg
    14281 gcagccggtg cgccgtcgat taggaagccg cccaagggcg acgagcaacc agattttttc
    14341 gttccgatgc tctatgacgt gggcacccgc gatagtcgca gcatcatgga cgtggccgtt
    14401 ttccgtctgt cgaagcgtga ccgacgagct ggcgaggtga tccgctacga gcttccagac
    14461 gggcacgtag aggtttccgc agggccggcc ggcatggcca gtgtgtggga ttacgacctg
    14521 gtactgatgg cggtttccca tctaaccgaa tccatgaacc gataccggga agggaaggga
    14581 gacaagcccg gccgcgtgtt ccgtccacac gttgcggacg tactcaagtt ctgccggcga
    14641 gccgatggcg gaaagcagaa agacgacctg gtagaaacct gcattcggtt aaacaccacg
    14701 cacgttgcca tgcagcgtac gaagaaggcc aagaacggcc gcctggtgac ggtatccgag
    14761 ggtgaagcct tgattagccg ctacaagatc gtaaagagcg aaaccgggcg gccggagtac
    14821 atcgagatcg agctagctga ttggatgtac cgcgagatca cagaaggcaa gaacccggac
    14881 gtgctgacgg ttcaccccga ttactttttg atcgatcccg gcatcggccg ttttctctac
    14941 cgcctggcac gccgcgccgc aggcaaggca gaagccagat ggttgttcaa gacgatctac
    15001 gaacgcagtg gcagcgccgg agagttcaag aagttctgtt tcaccgtgcg caagctgatc
    15061 gggtcaaatg acctgccgga gtacgatttg aaggaggagg cggggcaggc tggcccgatc
    15121 ctagtcatgc gctaccgcaa cctgatcgag ggcgaagcat ccgccggttc ctaatgtacg
    15181 gagcagatgc tagggcaaat tgccctagca ggggaaaaag gtcgaaaagg cctctttcct
    15241 gtggatagca cgtacattgg gaacccaaag ccgtacattg ggaaccggaa cccgtacatt
    15301 gggaacccaa agccgtacat tgggaaccgg tcacacatgt aagtgactga tataaaagag
    15361 aaaaaaggcg atttttccgc ctaaaactct ttaaaactta ttaaaactct taaaacccgc
    15421 ctggcctgtg cataactgtc tggccagcgc acagccgaag agctgcaaaa agcgcctacc
    15481 cttcggtcgc tgcgctccct acgccccgcc gcttcgcgtc ggcctatcgc ggccgctggc
    15541 cgctcaaaaa tggctggcct acggccaggc aatctaccag ggcgcggaca agccgcgccg
    15601 tcgccactcg accgccggcg cccacatcaa ggcaccctgc ctcgcgcgtt tcggtgatga
    15661 cggtgaaaac ctctgacaca tgcagctccc ggaaacggtc acagcttgtc tgtaagcgga
    15721 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc
    15781 agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca
    15841 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
    15901 agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
    15961 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
    16021 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
    16081 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
    16141 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
    16201 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
    16261 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
    16321 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
    16381 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
    16441 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
    16501 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
    16561 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
    16621 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
    16681 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
    16741 aactcacgtt aagggatttt ggtcatgcat tctaggtact aaaacaattc atccagtaaa
    16801 atataatatt ttattttctc ccaatcaggc ttgatcccca gtaagtcaaa aaatagctcg
    16861 acatactgtt cttccccgat atcctccctg atcgaccgga cgcagaaggc aatgtcatac
    16921 cacttgtccg ccctgccgct tctcccaaga tcaataaagc cacttacttt gccatctttc
    16981 acaaagatgt tgctgtctcc caggtcgccg tgggaaaaga caagttcctc ttcgggcttt
    17041 tccgtcttta aaaaatcata cagctcgcgc ggatctttaa atggagtgtc ttcttcccag
    17101 ttttcgcaat ccacatcggc cagatcgtta ttcagtaagt aatccaattc ggctaagcgg
    17161 ctgtctaagc tattcgtata gggacaatcc gatatgtcga tggagtgaaa gagcctgatg
    17221 cactccgcat acagctcgat aatcttttca gggctttgtt catcttcata ctcttccgag
    17281 caaaggacgc catcggcctc actcatgagc agattgctcc agccatcatg ccgttcaaag
    17341 tgcaggacct ttggaacagg cagctttcct tccagccata gcatcatgtc cttttcccgt
    17401 tccacatcat aggtggtccc tttataccgg ctgtccgtca tttttaaata taggttttca
    17461 ttttctccca ccagcttata taccttagca ggagacattc cttccgtatc ttttacgcag
    17521 cggtattttt cgatcagttt tttcaattcc ggtgatattc tcattttagc catttattat
    17581 ttccttcctc ttttctacag tatttaaaga taccccaaga agctaattat aacaagacga
    17641 actccaattc actgttcctt gcattctaaa accttaaata ccagaaaaca gctttttcaa
    17701 agttgttttc aaagttggcg tataacatag tatcgacgga gccgattttg aaaccgcggt
    17761 gatcacaggc agcaacgctc tgtcatcgtt acaatcaaca tgctaccctc cgcgagatca
    17821 tccgtgtttc aaacccggca gcttagttgc cgttcttccg aatagcatcg gtaacatgag
    17881 caaagtctgc cgccttacaa cggctctccc gctgacgccg tcccggactg atgggctgcc
    17941 tgtatcgagt ggtgattttg tgccgagctg ccggtcgggg agctgttggc tggctgg
//