pDicAID_nCas9-PmCDA-2A-NptII_ETR

LOCUS       pDicAID_nCas9-PmCDA-2A-NptII_ETR               16623 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  Dicot Target-AID vector expressing dicot-optimized 
            nCas9-PmCDA1-2A-NptII with sgRNA targeting SlEtr.
ACCESSION   .
VERSION     .
KEYWORDS    pDicAID_nCas9-PmCDA-2A-NptII_ETR
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16623)
  AUTHORS   Shimatani Z, Kashojiya S, Takayama M, Terada R, Arazoe T, Ishii H, 
            Teramura H, Yamamoto T, Komatsu H, Miura K, Ezura H, Nishida K, 
            Ariizumi T, Kondo A
  TITLE     Targeted base editing in rice and tomato using a CRISPR-Cas9 
            cytidine deaminase fusion.
  JOURNAL   Nat Biotechnol. 2017 Mar 27. doi: 10.1038/nbt.3833.
  PUBMED    28346401
REFERENCE   2  (bases 1 to 16623)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..16623
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    113..137
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     protein_bind    295..316
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        331..361
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    369..385
                     /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     374..396
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     393..409
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     393..409
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    485..509
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(559..575)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(560..576)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     protein_bind    596..695
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_RNA        1104..1179
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(1199..1223)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1229..1328)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1451..1575)
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2513..2560
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     intron          2561..2749
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     CDS             2765..6865
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6878..6898
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7091..7156
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             7163..7786
                     /codon_start=1
                     /product="cytidine deaminase from the sea lamprey 
                     Petromyzon marinus (Rogozin et al., 2007)"
                     /label=PmCDA1
                     /translation="MTDAEYVRIHEKLDIYTFKKQFFNNKKSVSHRCYVLFELKRRGER
                     RACFWGYAVNKPQSGTERGIHAEIFSIRKVEEYLRDNPGQFTINWYSSWSPCADCAEKI
                     LEWYNQELRGNGHTLKIWACKLYYEKNARNQIGLWNLRDNGVGLNVMVSEHYQCCRKIF
                     IQSSHNQLNENRWLEKTLKRAEKRRSELSIMIQVKILHTTKSPAV"
     CDS             7790..7810
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7883..8680
                     /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(7940..7959)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8550..8569
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     protein_bind    complement(9832..9956)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(9973..9990)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9973..9989)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9982..10004)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    10204..10228
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             10749..11540
                     /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      11786..12374
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12275..12294
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12528..12545
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    12560..12700
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(12626..12645)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     12786..12808
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      13044..13238
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(13304..14377)
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     CDS             complement(14806..15435)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
ORIGIN
        1 gatcccgagg ggaaccctgt ggttggcatg cacatacaaa tggacgaacg gataaacctt
       61 ttcacgccct tttaaatatc cgattattct aataaacgct cttttctctt aggtttaccc
      121 gccaatatat cctgtcaaac actgatagtt taaactgaag gcgggaaacg acaatctgat
      181 ccaagctcaa gctccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
      241 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
      301 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
      361 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgaca tgattacgaa
      421 tttcaggtga ctgatagtga cctgttcgtt gcaacaaatt gatgagcaat gcttttttat
      481 aatgccaact ttgtacaaaa aagcaggcgg cgcgtcctgc aggataggga taacagggta
      541 atggcgcgcc aagcttatcg ataccgtcga cctcgagcgg ccgcgggaat tcgatcaaat
      601 aatgatttta ttttgactga tagtgacctg ttcgttgcaa caaattgatg agcaatgctt
      661 ttttataatg ccaactttgt acaaaaaagc aggctctttt tttcttcttc ttcgttcata
      721 cagttttttt ttgtttatca gcttacattt tcttgaaccg tagctttcgt tttcttcttt
      781 ttaactttcc attcggagtt tttgtatctt gtttcatagt ttgtcccagg attagaatga
      841 ttaggcatcg aaccttcaag aatttgattg aataaaacat cttcattctt aagatatgaa
      901 gataatcttc aaaaggcccc tgggaatctg aaagaagaga agcaggccca tttatatggg
      961 aaagaacaat agtatttctt atataggccc atttaagttg aaaacaatct tcaaaagtcc
     1021 cacatcgctt agataagaaa acgaagctga gtttatatac agctagagtc gaagtagtga
     1081 ttgtgcacaa gaacccatct atagttttag agctagaaat agcaagttaa aataaggcta
     1141 gtccgttatc aacttgaaaa agtggcaccg agtcggtgct ttttttacgc gtttctagac
     1201 ccagctttct tgtacaaagt tggcattaac ccagctttct tgtacaaagt tggcattata
     1261 aaaaataatt gctcatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc
     1321 attatttgat cactagtgga tccgagctcg gtaccttaat taaattaccc tgttatccct
     1381 atcctgcagg attaacaccc aacttttcta tggcccctac aggtcactaa taccatctaa
     1441 gtagttgatt catagtgact ggatatgttg tgttttacag tattatgtag tctgtttttt
     1501 atgcaaaatc taatttaata tattgatatt tatatcattt tacgtttctc gttcaacttt
     1561 tctatacaaa gttgaagatc tgactagcaa cgattgtaca attgcttctt taaaaaagga
     1621 agaaagaaag aaagaaaaga atcaacatca gcgttaacaa acggccccgt tacggcccaa
     1681 acggtcatat agagtaacgg cgttaagcgt tgaaagactc ctatcgaaat acgtaaccgc
     1741 aaacgtgtca tagtcagatc ccctcttcct tcaccgcctc aaacacaaaa ataatcttct
     1801 acagcctata tatacaaccc ccccttctat ctctcctttc tcacaattca tcatctttct
     1861 ttctctaccc ccaattttaa gaaatcctct cttctcctct tcattttcaa ggtaaatctc
     1921 tctctctctc tctctctctg ttattccttg ttttaattag gtatgtatta ttgctagttt
     1981 gttaatctgc ttatcttatg tatgccttat gtgaatatct ttatcttgtt catctcatcc
     2041 gtttagaagc tataaatttg ttgatttgac tgtgtatcta cacgtggtta tgtttatatc
     2101 taatcagata tgaatttctt catattgttg cgtttgtgtg taccaatccg aaatcgttga
     2161 tttttttcat ttaatcgtgt agctaattgt acgtatacat atggatctac gtatcaattg
     2221 ttcatctgtt tgtgtttgta tgtatacaga tctgaaaaca tcacttctct catctgattg
     2281 tgttgttaca tacatagata tagatctgtt atatcatttt ttttattaat tgtgtatata
     2341 tatatgtgca tagatctgga ttacatgatt gtgattattt acatgatttt gttatttacg
     2401 tatgtatata tgtagatctg gactttttgg agttgttgac ttgattgtat ttgtgtgtgt
     2461 atatgtgtgt tctgatcttg atatgttatg tatgtgcagc gaattcacca tgaagcgtcc
     2521 agctgctacc aagaaagctg gacaggctaa gaagaagaag gtaagtttct gcttctacct
     2581 ttgatatata tataataatt atcattaatt agtagtaata taatatttca aatatttttt
     2641 tcaaaataaa agaatgtagt atatagcaat tgcttttctg tagtttataa gtgtgtatat
     2701 tttaatttat aacttttcta atatatgacc aaaatttgtt gatgtgcagg gccggcgcgc
     2761 aacggataag aagtactcta tcggactcgc tatcggaact aactctgtgg gatgggctgt
     2821 gatcaccgat gagtacaagg tgccatctaa gaagttcaag gttctcggaa acaccgatag
     2881 gcactctatc aagaaaaacc ttatcggtgc tctcctcttc gattctggtg aaactgctga
     2941 ggctaccaga ctcaagagaa ccgctagaag aaggtacacc agaagaaaga acaggatctg
     3001 ctacctccaa gagatcttct ctaacgagat ggctaaagtg gatgattcat tcttccacag
     3061 gctcgaagag tcattcctcg tggaagaaga taagaagcac gagaggcacc ctatcttcgg
     3121 aaacatcgtt gatgaggtgg cataccacga gaagtaccct actatctacc acctcagaaa
     3181 gaagctcgtt gattctactg ataaggctga tctcaggctc atctacctcg ctctcgctca
     3241 catgatcaag ttcagaggac acttcctcat cgagggtgat ctcaaccctg ataactctga
     3301 tgtggataag ttgttcatcc agctcgtgca gacctacaac cagcttttcg aagagaaccc
     3361 tatcaacgct tcaggtgtgg atgctaaggc tatcctctct gctaggctct ctaagtcaag
     3421 aaggcttgag aacctcattg ctcagctccc tggtgagaag aagaacggac ttttcggaaa
     3481 cttgatcgct ctctctctcg gactcacccc taacttcaag tctaacttcg atctcgctga
     3541 ggatgcaaag ctccagctct caaaggatac ctacgatgat gatctcgata acctcctcgc
     3601 tcagatcgga gatcagtacg ctgatttgtt cctcgctgct aagaacctct ctgatgctat
     3661 cctcctcagt gatatcctca gagtgaacac cgagatcacc aaggctccac tctcagcttc
     3721 tatgatcaag agatacgatg agcaccacca ggatctcaca cttctcaagg ctcttgttag
     3781 acagcagctc ccagagaagt acaaagagat tttcttcgat cagtctaaga acggatacgc
     3841 tggttacatc gatggtggtg catctcaaga agagttctac aagttcatca agcctatcct
     3901 cgagaagatg gatggaaccg aggaactcct cgtgaagctc aatagagagg atcttctcag
     3961 aaagcagagg accttcgata acggatctat ccctcatcag atccacctcg gagagttgca
     4021 cgctatcctt agaaggcaag aggatttcta cccattcctc aaggataaca gggaaaagat
     4081 tgagaagatt ctcaccttca gaatccctta ctacgtggga cctctcgcta gaggaaactc
     4141 aagattcgct tggatgacca gaaagtctga ggaaaccatc accccttgga acttcgaaga
     4201 ggtggtggat aagggtgcta gtgctcagtc tttcatcgag aggatgacca acttcgataa
     4261 gaaccttcca aacgagaagg tgctccctaa gcactctttg ctctacgagt acttcaccgt
     4321 gtacaacgag ttgaccaagg ttaagtacgt gaccgaggga atgaggaagc ctgctttttt
     4381 gtcaggtgag caaaagaagg ctatcgttga tctcttgttc aagaccaaca gaaaggtgac
     4441 cgtgaagcag ctcaaagagg attacttcaa gaaaatcgag tgcttcgatt cagttgagat
     4501 ttctggtgtt gaggataggt tcaacgcatc tctcggaacc taccacgatc tcctcaagat
     4561 cattaaggat aaggatttct tggataacga ggaaaacgag gatatcttgg aggatatcgt
     4621 tcttaccctc accctctttg aagatagaga gatgattgaa gaaaggctca agacctacgc
     4681 tcatctcttc gatgataagg tgatgaagca gttgaagaga agaagataca ctggttgggg
     4741 aaggctctca agaaagctca ttaacggaat cagggataag cagtctggaa agacaatcct
     4801 tgatttcctc aagtctgatg gattcgctaa cagaaacttc atgcagctca tccacgatga
     4861 ttctctcacc tttaaagagg atatccagaa ggctcaggtt tcaggacagg gtgatagtct
     4921 ccatgagcat atcgctaacc tcgctggatc tcctgcaatc aagaagggaa tcctccagac
     4981 tgtgaaggtt gtggatgagt tggtgaaggt gatgggaagg cataagcctg agaacatcgt
     5041 gatcgaaatg gctagagaga accagaccac tcagaaggga cagaagaact ctagggaaag
     5101 gatgaagagg atcgaggaag gtatcaaaga gcttggatct cagatcctca aagagcaccc
     5161 tgttgagaac actcagctcc agaatgagaa gctctacctc tactacctcc agaacggaag
     5221 ggatatgtat gtggatcaag agttggatat caacaggctc tctgattacg atgttgatca
     5281 tatcgtgcca cagtcattct tgaaggatga ttctatcgat aacaaggtgc tcaccaggtc
     5341 tgataagaac aggggtaaga gtgataacgt gccaagtgaa gaggttgtga agaaaatgaa
     5401 gaactattgg aggcagctcc tcaacgctaa gctcatcact cagagaaagt tcgataactt
     5461 gactaaggct gagaggggag gactctctga attggataag gcaggattca tcaagaggca
     5521 gcttgtggaa accaggcaga tcactaagca cgttgcacag atcctcgatt ctaggatgaa
     5581 caccaagtac gatgagaacg ataagttgat cagggaagtg aaggttatca ccctcaagtc
     5641 aaagctcgtg tctgatttca gaaaggattt ccaattctac aaggtgaggg aaatcaacaa
     5701 ctaccaccac gctcacgatg cttaccttaa cgctgttgtt ggaaccgctc tcatcaagaa
     5761 gtatcctaag ctcgagtcag agttcgtgta cggtgattac aaggtgtacg atgtgaggaa
     5821 gatgatcgct aagtctgagc aagagatcgg aaaggctacc gctaagtatt tcttctactc
     5881 taacatcatg aatttcttca agaccgagat taccctcgct aacggtgaga tcagaaagag
     5941 gccactcatc gagacaaacg gtgaaacagg tgagatcgtg tgggataagg gaagggattt
     6001 cgctaccgtt agaaaggtgc tctctatgcc acaggtgaac atcgttaaga aaaccgaggt
     6061 gcagaccggt ggattctcta aagagtctat cctccctaag aggaactctg ataagctcat
     6121 tgctaggaag aaggattggg accctaagaa atacggtggt ttcgattctc ctaccgtggc
     6181 ttactctgtt ctcgttgtgg ctaaggttga gaagggaaag agtaagaagc tcaagtctgt
     6241 taaggaactt ctcggaatca ctatcatgga aaggtcatct ttcgagaaga acccaatcga
     6301 tttcctcgag gctaagggat acaaagaggt taagaaggat ctcatcatca agctcccaaa
     6361 gtactcactc ttcgaactcg agaacggtag aaagaggatg ctcgcttctg ctggtgagct
     6421 tcaaaaggga aacgagcttg ctctcccatc taagtacgtt aactttcttt acctcgcttc
     6481 tcactacgag aagttgaagg gatctccaga agataacgag cagaagcaac ttttcgttga
     6541 gcagcacaag cactacttgg atgagatcat cgagcagatc tctgagttct ctaaaagggt
     6601 gatcctcgct gatgcaaacc tcgataaggt gttgtctgct tacaacaagc acagagataa
     6661 gcctatcagg gaacaggcag agaacatcat ccatctcttc acccttacca acctcggtgc
     6721 tcctgctgct ttcaagtact tcgatacaac catcgatagg aagagataca cctctaccaa
     6781 agaagtgctc gatgctaccc tcatccatca gtctatcact ggactctacg agactaggat
     6841 cgatctctca cagctcggtg gtgattcaag ggctgatcct aagaagaaga ggaaggttgg
     6901 aggagggcca ggagcagagt atgttagagc gttgtttgat tttaacggta acgatgaaga
     6961 ggatttaccc tttaagaaag gcgacattct caggattagg gataaacctg aagagcaatg
     7021 gtggaatgct gaggatagtg aaggcaaacg aggaatgatt ttagtgccgt atgtggagaa
     7081 atattcgggt gactacaaag atcatgatgg tgattacaaa gaccatgaca tcgactacaa
     7141 ggatgatgat gataagtcag ggatgacaga tgctgaatat gtcagaatcc acgaaaagtt
     7201 ggacatttac acgtttaaga agcagttctt caacaacaag aaatctgttt cgcataggtg
     7261 ctatgtgctt ttcgaactaa aacgtcgtgg agaaagacgg gcttgctttt ggggttacgc
     7321 ggttaacaaa ccacaatcag gtactgaacg aggaatacac gctgaaatct tttctatccg
     7381 aaaggttgag gaatatctac gtgacaatcc tggacagttc actatcaatt ggtattctag
     7441 ctggtcacca tgtgcagatt gtgctgagaa gattctcgaa tggtacaatc aagagcttag
     7501 aggcaatgga catacattga aaatatgggc atgcaagctc tactacgaaa agaatgccag
     7561 aaaccaaatt gggctttgga acttgaggga taatggagtt gggcttaatg tcatggtttc
     7621 tgagcactat caatgttgtc ggaagatctt catacaaagt tcccataacc agttgaatga
     7681 gaacagatgg ttagagaaaa cccttaaaag agccgagaag agaagatccg aactgagcat
     7741 tatgatacag gtcaaaattc tgcataccac taagagtcca gctgtaggtc ctaagaagaa
     7801 acgtaaagta gggcccggac aacttctcaa ctttgacttg ctaaatggtg atgttgaatc
     7861 taatcctgga ccaggttcta gaatggggat tgaacaagat ggattgcacg caggttctcc
     7921 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     7981 tgatgccgcc gtgtttcggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     8041 cctgtccggt gccctgaatg aactccagga cgaggcagcg cggctatcgt ggctggccac
     8101 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     8161 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     8221 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     8281 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     8341 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     8401 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacacatg gcgatgcctg
     8461 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     8521 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     8581 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     8641 gcgcatcgcc ttctatcgcc ttctcgacga gttcttctga gagctctggg ctaatctaaa
     8701 acgatttatc tgtggcttca agtgtatcga tcacttatgt gaggtgtaat tactggtgtt
     8761 tttggtgtgc tctggttcct ttcaagtgtg ttgttgccgc tcgaactact ccgctatgta
     8821 aaacggtaaa acctgttgtc tcattatgaa agtgaactat attatgttct actactactc
     8881 tacttagtca attttcttca ccttgattag tgtaaatatg aatcttatat tcttatgtct
     8941 taagaaatta gcacatgtga agcctccaag tgcatatttt ctcgatcgcg agacgcacaa
     9001 tgcgtgagaa attcagctgg ttatactcaa atatattaat atatctagca gcagctcatg
     9061 gagattcagg aaacttggca tccctaatcc ctaccatttc cattcttccg agattgacag
     9121 ttcaatacaa gtacagtaat ctcctggtaa gtttcttatt aacttgacat gtagtagtaa
     9181 taatttgtac gtagcataga tacatagaca caaaaatgtc ctccccattg agctagccga
     9241 ttggagccga acaccgcagg aatgaattta cataatctgc aaagaataaa tggaatgtgc
     9301 ctccacagga aaaccagcgg cagtgtggcg ttttcaagag cagccgtaag tcgaagccta
     9361 ttctgaatcg tagaaatcac tggggcatgg tgtaattaca tccgactcca acatctgtac
     9421 caccctgtgc attgtaggcc gctcctctgg caaagaactt acacattgtt tagcaagaga
     9481 aagtagagca tccaaggtct caatctgcac tccctcacaa tatggatcga caatttccct
     9541 ctcccgattc tcaccgacca ggaaattcaa ctgccacaga gcaagtagat tatttcaaga
     9601 atacattaaa tcaattgaag gcatacgtaa ttcatatcag aaaactgtgg atatgaaatg
     9661 gaaggacata aaggtcatac atacccatcc aacaatgttc aatccctttt caataaatga
     9721 tgcatcagta ggtcgttttc cgcttagtat ttcaagtagc aaaactccaa aactgtagac
     9781 gtcagtcttt tcggtggctc tgccactttg catatactcc tgcaggtcga ccatagtgac
     9841 tggatatgtt gtgttttaca gtattatgta gtctgttttt tatgcaaaat ctaatttaat
     9901 atattgatat ttatatcatt ttacgtttct cgttcagctt tcttgtacaa agtggtgata
     9961 tcccagcttg gcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
    10021 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
    10081 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgct agagcaattc
    10141 ggcgttaatt cagtacatta aaaacgtccg caatgtgtta ttaagttgtc taagcgtcaa
    10201 tttgtttaca ccacaatata tcctgccacc agccagccaa cagctccccg accggcagct
    10261 cggcacaaaa tcaccactcg atacaggcag cccatcagtc cgggacggcg tcagcgggag
    10321 agccgttgta aggcggcaga ctttgctcat gttaccgatg ctattcggaa gaacggcaac
    10381 taagctgccg ggtttgaaac acggatgatc tcgcggaggg tagcatgttg attgtaacga
    10441 tgacagagcg ttgctgcctg tgatcaattc gggcacgaac ccagtggaca taagcctgtt
    10501 cggttcgtaa gctgtaatgc aagtagcgta tgcgctcacg caactggtcc agaaccttga
    10561 ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat ggcttgttat gactgttttt
    10621 ttggggtaca gtctatgcct cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat
    10681 gtttgatgtt atggagcagc aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt
    10741 taaacatcat gggggaagcg gtgatcgccg aagtatcgac tcaactatca gaggtagttg
    10801 gcgtcatcga gcgccatctc gaaccgacgt tgctggccgt acatttgtac ggctccgcag
    10861 tggatggcgg cctgaagcca cacagtgata ttgatttgct ggttacggtg accgtaaggc
    10921 ttgatgaaac aacgcggcga gctttgatca acgacctttt ggaaacttcg gcttcccctg
    10981 gagagagcga gattctccgc gctgtagaag tcaccattgt tgtgcacgac gacatcattc
    11041 cgtggcgtta tccagctaag cgcgaactgc aatttggaga atggcagcgc aatgacattc
    11101 ttgcaggtat cttcgagcca gccacgatcg acattgatct ggctatcttg ctgacaaaag
    11161 caagagaaca tagcgttgcc ttggtaggtc cagcggcgga ggaactcttt gatccggttc
    11221 ctgaacagga tctatttgag gcgctaaatg aaaccttaac gctatggaac tcgccgcccg
    11281 actgggctgg cgatgagcga aatgtagtgc ttacgttgtc ccgcatttgg tacagcgcag
    11341 taaccggcaa aatcgcgccg aaggatgtcg ctgccgactg ggcaatggag cgcctgccgg
    11401 cccagtatca gcccgtcata cttgaagcta gacaggctta tcttggacaa gaagaagatc
    11461 gcttggcctc gcgcgcagat cagttggaag aatttgtcca ctacgtgaaa ggcgagatca
    11521 ccaaggtagt cggcaaataa tgtctagcta gaaattcgtt caagccgacg ccgcttcgcg
    11581 gcgcggctta actcaagcgt tagatgcact aagcacataa ttgctcacag ccaaactatc
    11641 aggtcaagtc tgcttttatt atttttaagc gtgcataata agccctacac aaattgggag
    11701 atatatcatg catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
    11761 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
    11821 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
    11881 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
    11941 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
    12001 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
    12061 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
    12121 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
    12181 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
    12241 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
    12301 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
    12361 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
    12421 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
    12481 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
    12541 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
    12601 cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
    12661 atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc
    12721 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
    12781 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca
    12841 gggtgccttg atgtgggcgc cggcggtcga gtggcgacgg cgcggcttgt ccgcgccctg
    12901 gtagattgcc tggccgtagg ccagccattt ttgagcggcc agcggccgcg ataggccgac
    12961 gcgaagcggc ggggcgtagg gagcgcagcg accgaagggt aggcgctttt tgcagctctt
    13021 cggctgtgcg ctggccagac agttatgcac aggccaggcg ggttttaaga gttttaataa
    13081 gttttaaaga gttttaggcg gaaaaatcgc cttttttctc ttttatatca gtcacttaca
    13141 tgtgtgaccg gttcccaatg tacggctttg ggttcccaat gtacgggttc cggttcccaa
    13201 tgtacggctt tgggttccca atgtacgtgc tatccacagg aaagagacct tttcgacctt
    13261 tttcccctgc tagggcaatt tgccctagca tctgctccgt acattaggaa ccggcggatg
    13321 cttcgccctc gatcaggttg cggtagcgca tgactaggat cgggccagcc tgccccgcct
    13381 cctccttcaa atcgtactcc ggcaggtcat ttgacccgat cagcttgcgc acggtgaaac
    13441 agaacttctt gaactctccg gcgctgccac tgcgttcgta gatcgtcttg aacaaccatc
    13501 tggcttctgc cttgcctgcg gcgcggcgtg ccaggcggta gagaaaacgg ccgatgccgg
    13561 gatcgatcaa aaagtaatcg gggtgaaccg tcagcacgtc cgggttcttg ccttctgtga
    13621 tctcgcggta catccaatca gctagctcga tctcgatgta ctccggccgc ccggtttcgc
    13681 tctttacgat cttgtagcgg ctaatcaagg cttcaccctc ggataccgtc accaggcggc
    13741 cgttcttggc cttcttcgta cgctgcatgg caacgtgcgt ggtgtttaac cgaatgcagg
    13801 tttctaccag gtcgtctttc tgctttccgc catcggctcg ccggcagaac ttgagtacgt
    13861 ccgcaacgtg tggacggaac acgcggccgg gcttgtctcc cttcccttcc cggtatcggt
    13921 tcatggattc ggttagatgg gaaaccgcca tcagtaccag gtcgtaatcc cacacactgg
    13981 ccatgccggc cggccctgcg gaaacctcta cgtgcccgtc tggaagctcg tagcggatca
    14041 cctcgccagc tcgtcggtca cgcttcgaca gacggaaaac ggccacgtcc atgatgctgc
    14101 gactatcgcg ggtgcccacg tcatagagca tcggaacgaa aaaatctggt tgctcgtcgc
    14161 ccttgggcgg cttcctaatc gacggcgcac cggctgccgg cggttgccgg gattctttgc
    14221 ggattcgatc agcggccgct tgccacgatt caccggggcg tgcttctgcc tcgatgcgtt
    14281 gccgctgggc ggcctgcgcg gccttcaact tctccaccag gtcatcaccc agcgccgcgc
    14341 cgatttgtac cgggccggat ggtttgcgac cgctcacgcc gattcctcgg gcttgggggt
    14401 tccagtgcca ttgcagggcc ggcagacaac ccagccgctt acgcctggcc aaccgcccgt
    14461 tcctccacac atggggcatt ccacggcgtc ggtgcctggt tgttcttgat tttccatgcc
    14521 gcctccttta gccgctaaaa ttcatctact catttattca tttgctcatt tactctggta
    14581 gctgcgcgat gtattcagat agcagctcgg taatggtctt gccttggcgt accgcgtaca
    14641 tcttcagctt ggtgtgatcc tccgccggca actgaaagtt gacccgcttc atggctggcg
    14701 tgtctgccag gctggccaac gttgcagcct tgctgctgcg tgcgctcgga cggccggcac
    14761 ttagcgtgtt tgtgcttttg ctcattttct ctttacctca ttaactcaaa tgagttttga
    14821 tttaatttca gcggccagcg cctggacctc gcgggcagcg tcgccctcgg gttctgattc
    14881 aagaacggtt gtgccggcgg cggcagtgcc tgggtagctc acgcgctgcg tgatacggga
    14941 ctcaagaatg ggcagctcgt acccggccag cgcctcggca acctcaccgc cgatgcgcgt
    15001 gcctttgatc gcccgcgaca cgacaaaggc cgcttgtagc cttccatccg tgacctcaat
    15061 gcgctgctta accagctcca ccaggtcggc ggtggcccat atgtcgtaag ggcttggctg
    15121 caccggaatc agcacgaagt cggctgcctt gatcgcggac acagccaagt ccgccgcctg
    15181 gggcgctccg tcgatcacta cgaagtcgcg ccggccgatg gccttcacgt cgcggtcaat
    15241 cgtcgggcgg tcgatgccga caacggttag cggttgatct tcccgcacgg ccgcccaatc
    15301 gcgggcactg ccctggggat cggaatcgac taacagaaca tcggccccgg cgagttgcag
    15361 ggcgcgggct agatgggttg cgatggtcgt cttgcctgac ccgcctttct ggttaagtac
    15421 agcgataacc ttcatgcgtt ccccttgcgt atttgtttat ttactcatcg catcatatac
    15481 gcagcgaccg catgacgcaa gctgttttac tcaaatacac atcacctttt tagacggcgg
    15541 cgctcggttt cttcagcggc caagctggcc ggccaggccg ccagcttggc atcagacaaa
    15601 ccggccagga tttcatgcag ccgcacggtt gagacgtgcg cgggcggctc gaacacgtac
    15661 ccggccgcga tcatctccgc ctcgatctct tcggtaatga aaaacggttc gtcctggccg
    15721 tcctggtgcg gtttcatgct tgttcctctt ggcgttcatt ctcggcggcc gccagggcgt
    15781 cggcctcggt caatgcgtcc tcacggaagg caccgcgccg cctggcctcg gtgggcgtca
    15841 cttcctcgct gcgctcaagt gcgcggtaca gggtcgagcg atgcacgcca agcagtgcag
    15901 ccgcctcttt cacggtgcgg ccttcctggt cgatcagctc gcgggcgtgc gcgatctgtg
    15961 ccggggtgag ggtagggcgg gggccaaact tcacgcctcg ggccttggcg gcctcgcgcc
    16021 cgctccgggt gcggtcgatg attagggaac gctcgaactc ggcaatgccg gcgaacacgg
    16081 tcaacaccat gcggccggcc ggcgtggtgg tgtcggccca cggctctgcc aggctacgca
    16141 ggcccgcgcc ggcctcctgg atgcgctcgg caatgtccag taggtcgcgg gtgctgcggg
    16201 ccaggcggtc tagcctggtc actgtcacaa cgtcgccagg gcgtaggtgg tcaagcatcc
    16261 tggccagctc cgggcggtcg cgcctggtgc cggtgatctt ctcggaaaac agcttggtgc
    16321 agccggccgc gtgcagttcg gcccgttggt tggtcaagtc ctggtcgtcg gtgctgacgc
    16381 gggcatagcc cagcaggcca gcggcggcgc tcttgttcat ggcgtaatgt ctccggttct
    16441 agtcgcaagt attctacttt atgcgactaa aacacgcgac aagaaaacgc caggaaaagg
    16501 gcagggcggc agcctgtcgc gtaacttagg acttgtgcga catgtcgttt tcagaagacg
    16561 gctgcactga acgtcagaag ccgactgcac tatagcagcg gaggggttgg atcaaagtac
    16621 ttt
//