pK7WGF2::hCas9

LOCUS       pK7WGF2::hCas9              14400 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  Expesses the human codon usage Cas9 nuclease (Mali et al. Science 
            339, 823-826, 2013) with an N-terminal GFP tag from the 35S promoter
            in the plant tissue .
ACCESSION   .
VERSION     .
KEYWORDS    pK7WGF2::hCas9
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14400)
  AUTHORS   Nekrasov V, Staskawicz B, Weigel D, Jones JD, Kamoun S
  TITLE     Targeted mutagenesis in the model plant Nicotiana benthamiana using 
            Cas9 RNA-guided endonuclease.
  JOURNAL   Nat Biotechnol. 2013 Aug 8;31(8):691-3. doi: 10.1038/nbt.2655.
  PUBMED    23929340
REFERENCE   2  (bases 1 to 14400)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..14400
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             1..4104
                     /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             4117..4137
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(4156..4180)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        4552..4731
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             4765..5559
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4819..4838)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5429..5448
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      5599..5851
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    5944..5968
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             6496..7287
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      7531..8119
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8020..8039
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8273..8290
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    8305..8445
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(8371..8390)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8531..8553
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      8789..8983
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(9049..10116)
                     /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"
     CDS             complement(10550..11179)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    12479..12503
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     promoter        13196..13540
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     13484..13504
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             13633..14349
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(13678..13699)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(13939..13958)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     14286..14307
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    14356..14380
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 atggacaaga agtactccat tgggctcgat atcggcacaa acagcgtcgg ctgggccgtc
       61 attacggacg agtacaaggt gccgagcaaa aaattcaaag ttctgggcaa taccgatcgc
      121 cacagcataa agaagaacct cattggcgcc ctcctgttcg actccgggga gacggccgaa
      181 gccacgcggc tcaaaagaac agcacggcgc agatataccc gcagaaagaa tcggatctgc
      241 tacctgcagg agatctttag taatgagatg gctaaggtgg atgactcttt cttccatagg
      301 ctggaggagt cctttttggt ggaggaggat aaaaagcacg agcgccaccc aatctttggc
      361 aatatcgtgg acgaggtggc gtaccatgaa aagtacccaa ccatatatca tctgaggaag
      421 aagcttgtag acagtactga taaggctgac ttgcggttga tctatctcgc gctggcgcat
      481 atgatcaaat ttcggggaca cttcctcatc gagggggacc tgaacccaga caacagcgat
      541 gtcgacaaac tctttatcca actggttcag acttacaatc agcttttcga agagaacccg
      601 atcaacgcat ccggagttga cgccaaagca atcctgagcg ctaggctgtc caaatcccgg
      661 cggctcgaaa acctcatcgc acagctccct ggggagaaga agaacggcct gtttggtaat
      721 cttatcgccc tgtcactcgg gctgaccccc aactttaaat ctaacttcga cctggccgaa
      781 gatgccaagc ttcaactgag caaagacacc tacgatgatg atctcgacaa tctgctggcc
      841 cagatcggcg accagtacgc agaccttttt ttggcggcaa agaacctgtc agacgccatt
      901 ctgctgagtg atattctgcg agtgaacacg gagatcacca aagctccgct gagcgctagt
      961 atgatcaagc gctatgatga gcaccaccaa gacttgactt tgctgaaggc ccttgtcaga
     1021 cagcaactgc ctgagaagta caaggaaatt ttcttcgatc agtctaaaaa tggctacgcc
     1081 ggatacattg acggcggagc aagccaggag gaattttaca aatttattaa gcccatcttg
     1141 gaaaaaatgg acggcaccga ggagctgctg gtaaagctta acagagaaga tctgttgcgc
     1201 aaacagcgca ctttcgacaa tggaagcatc ccccaccaga ttcacctggg cgaactgcac
     1261 gctatcctca ggcggcaaga ggatttctac ccctttttga aagataacag ggaaaagatt
     1321 gagaaaatcc tcacatttcg gataccctac tatgtaggcc ccctcgcccg gggaaattcc
     1381 agattcgcgt ggatgactcg caaatcagaa gagaccatca ctccctggaa cttcgaggaa
     1441 gtcgtggata agggggcctc tgcccagtcc ttcatcgaaa ggatgactaa ctttgataaa
     1501 aatctgccta acgaaaaggt gcttcctaaa cactctctgc tgtacgagta cttcacagtt
     1561 tataacgagc tcaccaaggt caaatacgtc acagaaggga tgagaaagcc agcattcctg
     1621 tctggagagc agaagaaagc tatcgtggac ctcctcttca agacgaaccg gaaagttacc
     1681 gtgaaacagc tcaaagaaga ctatttcaaa aagattgaat gtttcgactc tgttgaaatc
     1741 agcggagtgg aggatcgctt caacgcatcc ctgggaacgt atcacgatct cctgaaaatc
     1801 attaaagaca aggacttcct ggacaatgag gagaacgagg acattcttga ggacattgtc
     1861 ctcaccctta cgttgtttga agatagggag atgattgaag aacgcttgaa aacttacgct
     1921 catctcttcg acgacaaagt catgaaacag ctcaagaggc gccgatatac aggatggggg
     1981 cggctgtcaa gaaaactgat caatgggatc cgagacaagc agagtggaaa gacaatcctg
     2041 gattttctta agtccgatgg atttgccaac cggaacttca tgcagttgat ccatgatgac
     2101 tctctcacct ttaaggagga catccagaaa gcacaagttt ctggccaggg ggacagtctt
     2161 cacgagcaca tcgctaatct tgcaggtagc ccagctatca aaaagggaat actgcagacc
     2221 gttaaggtcg tggatgaact cgtcaaagta atgggaaggc ataagcccga gaatatcgtt
     2281 atcgagatgg cccgagagaa ccaaactacc cagaagggac agaagaacag tagggaaagg
     2341 atgaagagga ttgaagaggg tataaaagaa ctggggtccc aaatccttaa ggaacaccca
     2401 gttgaaaaca cccagcttca gaatgagaag ctctacctgt actacctgca gaacggcagg
     2461 gacatgtacg tggatcagga actggacatc aatcggctct ccgactacga cgtggatcat
     2521 atcgtgcccc agtcttttct caaagatgat tctattgata ataaagtgtt gacaagatcc
     2581 gataaaaata gagggaagag tgataacgtc ccctcagaag aagttgtcaa gaaaatgaaa
     2641 aattattggc ggcagctgct gaacgccaaa ctgatcacac aacggaagtt cgataatctg
     2701 actaaggctg aacgaggtgg cctgtctgag ttggataaag ccggcttcat caaaaggcag
     2761 cttgttgaga cacgccagat caccaagcac gtggcccaaa ttctcgattc acgcatgaac
     2821 accaagtacg atgaaaatga caaactgatt cgagaggtga aagttattac tctgaagtct
     2881 aagctggtct cagatttcag aaaggacttt cagttttata aggtgagaga gatcaacaat
     2941 taccaccatg cgcatgatgc ctacctgaat gcagtggtag gcactgcact tatcaaaaaa
     3001 tatcccaagc ttgaatctga atttgtttac ggagactata aagtgtacga tgttaggaaa
     3061 atgatcgcaa agtctgagca ggaaataggc aaggccaccg ctaagtactt cttttacagc
     3121 aatattatga attttttcaa gaccgagatt acactggcca atggagagat tcggaagcga
     3181 ccacttatcg aaacaaacgg agaaacagga gaaatcgtgt gggacaaggg tagggatttc
     3241 gcgacagtcc ggaaggtcct gtccatgccg caggtgaaca tcgttaaaaa gaccgaagta
     3301 cagaccggag gcttctccaa ggaaagtatc ctcccgaaaa ggaacagcga caagctgatc
     3361 gcacgcaaaa aagattggga ccccaagaaa tacggcggat tcgattctcc tacagtcgct
     3421 tacagtgtac tggttgtggc caaagtggag aaagggaagt ctaaaaaact caaaagcgtc
     3481 aaggaactgc tgggcatcac aatcatggag cgatcaagct tcgaaaaaaa ccccatcgac
     3541 tttctcgagg cgaaaggata taaagaggtc aaaaaagacc tcatcattaa gcttcccaag
     3601 tactctctct ttgagcttga aaacggccgg aaacgaatgc tcgctagtgc gggcgagctg
     3661 cagaaaggta acgagctggc actgccctct aaatacgtta atttcttgta tctggccagc
     3721 cactatgaaa agctcaaagg gtctcccgaa gataatgagc agaagcagct gttcgtggaa
     3781 caacacaaac actaccttga tgagatcatc gagcaaataa gcgaattctc caaaagagtg
     3841 atcctcgccg acgctaacct cgataaggtg ctttctgctt acaataagca cagggataag
     3901 cccatcaggg agcaggcaga aaacattatc cacttgttta ctctgaccaa cttgggcgcg
     3961 cctgcagcct tcaagtactt cgacaccacc atagacagaa agcggtacac ctctacaaag
     4021 gaggtcctgg acgccacact gattcatcag tcaattacgg ggctctatga aacaagaatc
     4081 gacctctctc agctcggtgg agacagcagg gctgacccca agaagaagag gaaggtgtga
     4141 agggtgggcg cgccgaccca gctttcttgt acaaagtggt gatatcccgc ggccatgcta
     4201 gagtccgcaa aaatcaccag tctctctcta caaatctatc tctctctatt tttctccaga
     4261 ataatgtgtg agtagttccc agataaggga attagggttc ttatagggtt tcgctcatgt
     4321 gttgagcata taagaaaccc ttagtatgta tttgtatttg taaaatactt ctatcaataa
     4381 aatttctaat tcctaaaacc aaaatccagt gacctgcagg catgcgacgt cgggccctct
     4441 agaggatccc cgggtaccgc gaattatcga tcatgagcgg agaattaagg gagtcacgtt
     4501 atgacccccg ccgatgacgc gggacaagcc gttttacgtt tggaactgac agaaccgcaa
     4561 cgttgaagga gccactgagc cgcgggtttc tggagtttaa tgagctaagc acatacgtca
     4621 gaaaccatta ttgcgcgttc aaaagtcgcc taaggtcact atcagctagc aaatatttct
     4681 tgtcaaaaat gctccactga cgttccataa attcccctcg gtatccaatt agagtctcat
     4741 attcactctc aactcgatcg aggcatgatt gaacaagatg gattgcacgc aggttctccg
     4801 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4861 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4921 ctgtccggtg ccctgaatga actccaagac gaggcagcgc ggctatcgtg gctggccacg
     4981 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     5041 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     5101 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     5161 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     5221 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     5281 aggctcaagg cgcggatgcc cgacggcgag gatctcgtcg tgacccacgg cgatgcctgc
     5341 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     5401 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     5461 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     5521 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcggac
     5581 tctagctaga gtcaagcaga tcgttcaaac atttggcaat aaagtttctt aagattgaat
     5641 cctgttgccg gtcttgcgat gattatcata taatttctgt tgaattacgt taagcatgta
     5701 ataattaaca tgtaatgcat gacgttattt atgagatggg tttttatgat tagagtcccg
     5761 caattataca tttaatacgc gatagaaaac aaaatatagc gcgcaaacta ggataaatta
     5821 tcgcgcgcgg tgtcatctat gttactagat cgaccggcat gcaagctgat aattcaattc
     5881 ggcgttaatt cagtacatta aaaacgtccg caatgtgtta ttaagttgtc taagcgtcaa
     5941 tttgtttaca ccacaatata tcctgccacc agccagccaa cagctccccg accggcagct
     6001 cggcacaaaa tcaccactcg atacaggcag cccatcagtc cgggacggcg tcagcgggag
     6061 agccgttgta aggcggcaga ctttgctcat gttaccgatg ctattcggaa gaacggcaac
     6121 taagctgccg ggtttgaaac acggatgatc tcgcggaggg tagcatgttg attgtaacga
     6181 tgacagagcg ttgctgcctg tgatcaattc gggcacgaac ccagtggaca taagcctcgt
     6241 tcggttcgta agctgtaatg caagtagcgt aactgccgtc acgcaactgg tccagaacct
     6301 tgaccgaacg cagcggtggt aacggcgcag tggcggtttt catggcttct tgttatgaca
     6361 tgtttttttg gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg
     6421 ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagggcag tcgccctaaa
     6481 acaaagttaa acatcatggg ggaagcggtg atcgccgaag tatcgactca actatcagag
     6541 gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc
     6601 tccgcagtgg atggcggcct gaagccacac agtgatattg atttgctggt tacggtgacc
     6661 gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga aacttcggct
     6721 tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt gcacgacgac
     6781 atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg gcagcgcaat
     6841 gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc tatcttgctg
     6901 acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga actctttgat
     6961 ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct atggaactcg
     7021 ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg catttggtac
     7081 agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc
     7141 ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct tggacaagaa
     7201 gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc
     7261 gagatcacca aggtagtcgg caaataatgt ctagctagaa attcgttcaa gccgacgccg
     7321 cttcgccggc gttaactcaa gcgattagat gcactaagca cataattgct cacagccaaa
     7381 ctatcaggtc aagtctgctt ttattatttt taagcgtgca taataagccc tacacaaatt
     7441 gggagatata tcatgcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     7501 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     7561 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     7621 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     7681 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     7741 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     7801 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     7861 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     7921 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     7981 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     8041 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     8101 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     8161 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     8221 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     8281 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     8341 tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     8401 ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca
     8461 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     8521 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     8581 aggcagggtg ccttgatgtg ggcgccggcg gtcgagtggc gacggcgcgg cttgtccgcg
     8641 ccctggtaga ttgcctggcc gtaggccagc catttttgag cggccagcgg ccgcgatagg
     8701 ccgacgcgaa gcggcggggc gtagggagcg cagcgaccga agggtaggcg ctttttgcag
     8761 ctcttcggct gtgcgctggc cagacagtta tgcacaggcc aggcgggttt taagagtttt
     8821 aataagtttt aaagagtttt aggcggaaaa atcgcctttt ttctctttta tatcagtcac
     8881 ttacatgtgt gaccggttcc caatgtacgg ctttgggttc ccaatgtacg ggttccggtt
     8941 cccaatgtac ggctttgggt tcccaatgta cgtgctatcc acaggaaaga gaccttttcg
     9001 acctttttcc cctgctaggg caatttgccc tagcatctgc tccgtacatt aggaaccggc
     9061 ggatgcttcg ccctcgatca ggttgcggta gcgcatgact aggatcgggc cagcctgccc
     9121 cgcctcctcc ttcaaatcgt actccggcag gtcatttgac ccgatcagct tgcgcacggt
     9181 gaaacagaac ttcttgaact ctccggcgct gccactgcgt tcgtagatcg tcttgaacaa
     9241 ccatctggct tctgccttgc ctgcggcgcg gcgtgccagg cggtagagaa aacggccgat
     9301 gccgggatcg atcaaaaagt aatcggggtg aaccgtcagc acgtccgggt tcttgccttc
     9361 tgtgatctcg cggtacatcc aatcagctag ctcgatctcg atgtactccg gccgcccggt
     9421 ttcgctcttt acgatcttgt agcggctaat caaggcttca ccctcggata ccgtcaccag
     9481 gcggccgttc ttggccttct tcgtacgctg catggcaacg tgcgtggtgt ttaaccgaat
     9541 gcaggtttct accaggtcgt ctttctgctt tccgccatcg gctcgccggc agaacttgag
     9601 tacgtccgca acgtgtggac ggaacacgcg gccgggcttg tctcccttcc cttcccggta
     9661 tcggttcatg gattcggtta gatgggaaac cgccatcagt accaggtcgt aatcccacac
     9721 actggccatg ccggccggcc ctgcggaaac ctctacgtgc ccgtctggaa gctcgtagcg
     9781 gatcacctcg ccagctcgtc ggtcacgctt cgacagacgg aaaacggcca cgtccatgat
     9841 gctgcgacta tcgcgggtgc ccacgtcata gagcatcgga acgaaaaaat ctggttgctc
     9901 gtcgcccttg ggcggcttcc taatcgacgg cgcaccggct gccggcggtt gccgggattc
     9961 tttgcggatt cgatcagcgg ccgcttgcca cgattcaccg gggcgtgctt ctgcctcgat
    10021 gcgttgccgc tgggcggcct gcgcggcctt caacttctcc accaggtcat cacccagcgc
    10081 cgcgccgatt tgtaccgggc cggatggttt gcgaccgtca cgccgattcc tcgggcttgg
    10141 gggttccagt gccattgcag ggccggcaga caacccagcc gcttacgcct ggccaaccgc
    10201 ccgttcctcc acacatgggg cattccacgg cgtcggtgcc tggttgttct tgattttcca
    10261 tgccgcctcc tttagccgct aaaattcatc tactcattta ttcatttgct catttactct
    10321 ggtagctgcg cgatgtattc agatagcagc tcggtaatgg tcttgccttg gcgtaccgcg
    10381 tacatcttca gcttggtgtg atcctccgcc ggcaactgaa agttgacccg cttcatggct
    10441 ggcgtgtctg ccaggctggc caacgttgca gccttgctgc tgcgtgcgct cggacggccg
    10501 gcacttagcg tgtttgtgct tttgctcatt ttctctttac ctcattaact caaatgagtt
    10561 ttgatttaat ttcagcggcc agcgcctgga cctcgcgggc agcgtcgccc tcgggttctg
    10621 attcaagaac ggttgtgccg gcggcggcag tgcctgggta gctcacgcgc tgcgtgatac
    10681 gggactcaag aatgggcagc tcgtacccgg ccagcgcctc ggcaacctca ccgccgatgc
    10741 gcgtgccttt gatcgcccgc gacacgacaa aggccgcttg tagccttcca tccgtgacct
    10801 caatgcgctg cttaaccagc tccaccaggt cggcggtggc ccatatgtcg taagggcttg
    10861 gctgcaccgg aatcagcacg aagtcggctg ccttgatcgc ggacacagcc aagtccgccg
    10921 cctggggcgc tccgtcgatc actacgaagt cgcgccggcc gatggccttc acgtcgcggt
    10981 caatcgtcgg gcggtcgatg ccgacaacgg ttagcggttg atcttcccgc acggccgccc
    11041 aatcgcgggc actgccctgg ggatcggaat cgactaacag aacatcggcc ccggcgagtt
    11101 gcagggcgcg ggctagatgg gttgcgatgg tcgtcttgcc tgacccgcct ttctggttaa
    11161 gtacagcgat aaccttcatg cgttcccctt gcgtatttgt ttatttactc atcgcatcat
    11221 atacgcagcg accgcatgac gcaagctgtt ttactcaaat acacatcacc tttttagacg
    11281 gcggcgctcg gtttcttcag cggccaagct ggccggccag gccgccagct tggcatcaga
    11341 caaaccggcc aggatttcat gcagccgcac ggttgagacg tgcgcgggcg gctcgaacac
    11401 gtacccggcc gcgatcatct ccgcctcgat ctcttcggta atgaaaaacg gttcgtcctg
    11461 gccgtcctgg tgcggtttca tgcttgttcc tcttggcgtt cattctcggc ggccgccagg
    11521 gcgtcggcct cggtcaatgc gtcctcacgg aaggcaccgc gccgcctggc ctcggtgggc
    11581 gtcacttcct cgctgcgctc aagtgcgcgg tacagggtcg agcgatgcac gccaagcagt
    11641 gcagccgcct ctttcacggt gcggccttcc tggtcgatca gctcgcgggc gtgcgcgatc
    11701 tgtgccgggg tgagggtagg gcgggggcca aacttcacgc ctcgggcctt ggcggcctcg
    11761 cgcccgctcc gggtgcggtc gatgattagg gaacgctcga actcggcaat gccggcgaac
    11821 acggtcaaca ccatgcggcc ggccggcgtg gtggtgtcgg cccacggctc tgccaggcta
    11881 cgcaggcccg cgccggcctc ctggatgcgc tcggcaatgt ccagtaggtc gcgggtgctg
    11941 cgggccaggc ggtctagcct ggtcactgtc acaacgtcgc cagggcgtag gtggtcaagc
    12001 atcctggcca gctccgggcg gtcgcgcctg gtgccggtga tcttctcgga aaacagcttg
    12061 gtgcagccgg ccgcgtgcag ttcggcccgt tggttggtca agtcctggtc gtcggtgctg
    12121 acgcgggcat agcccagcag gccagcggcg gcgctcttgt tcatggcgta atgtctccgg
    12181 ttctagtcgc aagtattcta ctttatgcga ctaaaacacg cgacaagaaa acgccaggaa
    12241 aagggcaggg cggcagcctg tcgcgtaact taggacttgt gcgacatgtc gttttcagaa
    12301 gacggctgca ctgaacgtca gaagccgact gcactatagc agcggagggg ttggatcaaa
    12361 gtactttgat cccgagggga accctgtggt tggcatgcac atacaaatgg acgaacggat
    12421 aaaccttttc acgccctttt aaatatccgt tattctaata aacgctcttt tctcttaggt
    12481 ttacccgcca atatatcctg tcaaacactg atagtttaaa ctgaaggcgg gaaacgacaa
    12541 tctgatccaa gctcaagcta agcttgagct ctcccatatg gtcgactaga gccaagctga
    12601 tctcctttgc cccggagatc accatggacg actttctcta tctctacgat ctaggaagaa
    12661 agttcgacgg agaaggtgac gataccatgt tcaccaccga taatgagaag attagcctct
    12721 tcaatttcag aaagaatgct gacccacaga tggttagaga ggcctacgcg gcaggtctca
    12781 tcaagacgat ctacccgagt aataatctcc aggagatcaa ataccttccc aagaaggtta
    12841 aagatgcagt caaaagattc aggactaact gcatcaagaa cacagagaaa gatatatttc
    12901 tcaagatcag aagtactatt ccagtatgga cgattcaagg cttgcttcat aaaccaaggc
    12961 aagtaataga gattggagtc tctaagaaag tagttcctac tgaatcaaag gccatggagt
    13021 caaaaattca gatcgaggat ctaacagaac tcgccgtgaa gactggcgaa cagttcatac
    13081 agagtctttt acgactcaat gacaagaaga aaatcttcgt caacatggtg gagcacgaca
    13141 ctctcgtcta ctccaagaat atcaaagata cagtctcaga agaccaaagg gctattgaga
    13201 cttttcaaca aagggtaata tcgggaaacc tcctcggatt ccattgccca gctatctgtc
    13261 acttcatcaa aaggacagta gaaaaggaag gtggcaccta caaatgccat cattgcgata
    13321 aaggaaaggc tatcgttcaa gatgcctctg ccgacagtgg tcccaaagat ggacccccac
    13381 ccacgaggag catcgtggaa aaagaagacg ttccaaccac gtcttcaaag caagtggatt
    13441 gatgtgatat ctccactgac gtaagggatg acgcacaatc ccactatcct tcgcaagacc
    13501 cttcctctat ataaggaagt tcatttcatt tggagaggac tccggtattt ttacaacaat
    13561 accacaacaa aacaaacaac aaacaacatt acaatttact attctagtcg acctgcaggc
    13621 ggccgcacta gtatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
    13681 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
    13741 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
    13801 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
    13861 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
    13921 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
    13981 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
    14041 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
    14101 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
    14161 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
    14221 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
    14281 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
    14341 ctgtacaagg atatcacaag tttgtacaaa aaagcaggct ccgcggccgc ccccttcacc
//