pHSN6I01

LOCUS       pHSN6I01               13088 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  CRISPR/Cas based plant genome editing and gene regulation; expresses
            dCas9-KRAB, gRNA scaffold for insertion of target sequence (AtU6-26 
            promoter), Hyg resistance.
ACCESSION   .
VERSION     .
KEYWORDS    pHSN6I01
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13088)
  AUTHORS   Xing HL, Dong L, Wang ZP, Zhang HY, Han CY, Liu B, Wang XC, Chen QJ
  TITLE     A CRISPR/Cas9 toolkit for multiplex genome editing in plants.
  JOURNAL   BMC Plant Biol. 2014 Nov 29;14(1):327.
  PUBMED    25432517
REFERENCE   2  (bases 1 to 13088)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..13088
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    1..25
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             810..1601
                     /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"
     misc_RNA        1753..1828
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2442..2786
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     2730..2750
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             2969..3034
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3041..3061
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3086..7186
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     RBS             7099..7107
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             7187..7234
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7253..7273
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7289..7309
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7346..7531
                     /codon_start=1
                     /gene="ZNF10"
                     /gene_synonym="KOX1"
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEP"
     CDS             7538..7558
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7565..7591
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     terminator      7644..7896
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        8012..8689
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     CDS             8757..9782
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     polyA_signal    9822..9996
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    10146..10170
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     complement(10418..10441)
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     terminator      10449..10476
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      10568..10654
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      10823..11258
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     rep_origin      complement(11373..11961)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11453..11472)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(12051..12860)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVSLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     12774..12793
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     12852..12871
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 gtttacccgc caatatatcc tgtcaaacac tgatagttta aactgaaggc gggaaacgac
       61 aatctgatcc aagctcaagc taagctcacg tgacggaatt aagcttcgac ttgccttccg
      121 cacaatacat catttcttct tagctttttt tcttcttctt cgttcataca gttttttttt
      181 gtttatcagc ttacattttc ttgaaccgta gctttcgttt tcttcttttt aactttccat
      241 tcggagtttt tgtatcttgt ttcatagttt gtcccaggat tagaatgatt aggcatcgaa
      301 ccttcaagaa tttgattgaa taaaacatct tcattcttaa gatatgaaga taatcttcaa
      361 aaggcccctg ggaatctgaa agaagagaag caggcccatt tatatgggaa agaacaatag
      421 tatttcttat ataggcccat ttaagttgaa aacaatcttc aaaagtccca catcgcttag
      481 ataagaaaac gaagctgagt ttatatacag ctagagtcga agtagtgatt gggagaccaa
      541 cccagtggac ataagcctgt tcggttcgta agctgtaatg caagtagcgt atgcgctcac
      601 gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg gcggttttca
      661 tggcttgtta tgactgtttt tttggggtac agtctatgcc tcgggcatcc aagcagcaag
      721 cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagg
      781 gcagtcgccc taaaacaaag ttaaacatca tgggggaagc ggtgatcgcc gaagtatcga
      841 ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg
      901 tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagtgat attgatttgc
      961 tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt
     1021 tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg
     1081 ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag
     1141 aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc
     1201 tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg
     1261 aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa
     1321 cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt
     1381 cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact
     1441 gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct agacaggctt
     1501 atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc
     1561 actacgtgaa aggcgagatc accaaggtag tcggcaaata atgtctagct agaaattcgt
     1621 tcaagccgac gccgcttcgc ggcgcggctt aactcaagcg ttagatgcac taagcacata
     1681 attgctcaca gccaaactat caggtcaagt ctgcttttat tatttttaag cgtgcataat
     1741 aagccggtct cggttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca
     1801 acttgaaaaa gtggcaccga gtcggtgctt ttttttgcaa aattttccag atcgatttct
     1861 tcttcctctg ttcttcggcg ttcaatttct ggggttttct cttcgttttc tgtaactgaa
     1921 acctaaaatt tgacctaaaa aaaatctcaa ataatatgat tcagtggttt tgtacttttc
     1981 agttagttga gttttgcagt tccgatgaga taaaccaata agcttgcatg cctgcaggtc
     2041 aacatggtgg agcacgacac acttgtctac tccaaaaata tcaaagatac agtctcagaa
     2101 gaccaaaggg caattgagac ttttcaacaa agggtaatat ccggaaacct cctcggattc
     2161 cattgcccag ctatctgtca ctttattgtg aagatagtgg aaaaggaagg tggctcctac
     2221 aaatgccatc attgcgataa aggaaaggcc atcgttgaag atgcctctgc cgacagtggt
     2281 cccaaagatg gacccccacc cacgaggagc atcgtggaaa aagaagacgt tccaaccacg
     2341 tcttcaaagc aagtggattg atgtgataac atggtggagc acgacacact tgtctactcc
     2401 aaaaatatca aagatacagt ctcagaagac caaagggcaa ttgagacttt tcaacaaagg
     2461 gtaatatccg gaaacctcct cggattccat tgcccagcta tctgtcactt tattgtgaag
     2521 atagtggaaa aggaaggtgg ctcctacaaa tgccatcatt gcgataaagg aaaggccatc
     2581 gttgaagatg cctctgccga cagtggtccc aaagatggac ccccacccac gaggagcatc
     2641 gtggaaaaag aagacgttcc aaccacgtct tcaaagcaag tggattgatg tgatatctcc
     2701 actgacgtaa gggatgacgc acaatcccac tatccttcgc aagacccttc ctctatataa
     2761 ggaagttcat ttcatttgga gaggacctcg acctcaacac aacatataca aaacaaacga
     2821 atctcaagca atcaagcatt ctacttctat tgcagcaatt taaatcattt cttttaaagc
     2881 aaaagcaatt ttctgaaaat tttcaccatt tacgaacgat actcgagggg gatccccaat
     2941 acttgtatgg ccgcggccgc tctagatgga ttacaaggac cacgacgggg attacaagga
     3001 ccacgacatt gattacaagg atgatgatga caagatggct ccgaagaaga agaggaaggt
     3061 tggcatccac ggggtgccag ctgctgacaa gaagtactcg atcggcctcg ccattgggac
     3121 taactctgtt ggctgggccg tgatcaccga cgagtacaag gtgccctcaa agaagttcaa
     3181 ggtcctgggc aacaccgatc ggcattccat caagaagaat ctcattggcg ctctcctgtt
     3241 cgacagcggc gagacggctg aggctacgcg gctcaagcgc accgcccgca ggcggtacac
     3301 gcgcaggaag aatcgcatct gctacctgca ggagattttc tccaacgaga tggcgaaggt
     3361 tgacgattct ttcttccaca ggctggagga gtcattcctc gtggaggagg ataagaagca
     3421 cgagcggcat ccaatcttcg gcaacattgt cgacgaggtt gcctaccacg agaagtaccc
     3481 tacgatctac catctgcgga agaagctcgt ggactccaca gataaggcgg acctccgcct
     3541 gatctacctc gctctggccc acatgattaa gttcaggggc catttcctga tcgaggggga
     3601 tctcaacccg gacaatagcg atgttgacaa gctgttcatc cagctcgtgc agacgtacaa
     3661 ccagctcttc gaggagaacc ccattaatgc gtcaggcgtc gacgcgaagg ctatcctgtc
     3721 cgctaggctc tcgaagtctc ggcgcctcga gaacctgatc gcccagctgc cgggcgagaa
     3781 gaagaacggc ctgttcggga atctcattgc gctcagcctg gggctcacgc ccaacttcaa
     3841 gtcgaatttc gatctcgctg aggacgccaa gctgcagctc tccaaggaca catacgacga
     3901 tgacctggat aacctcctgg cccagatcgg cgatcagtac gcggacctgt tcctcgctgc
     3961 caagaatctg tcggacgcca tcctcctgtc tgatattctc agggtgaaca ccgagattac
     4021 gaaggctccg ctctcagcct ccatgatcaa gcgctacgac gagcaccatc aggatctgac
     4081 cctcctgaag gcgctggtca ggcagcagct ccccgagaag tacaaggaga tcttcttcga
     4141 tcagtcgaag aacggctacg ctgggtacat tgacggcggg gcctctcagg aggagttcta
     4201 caagttcatc aagccgattc tggagaagat ggacggcacg gaggagctgc tggtgaagct
     4261 caatcgcgag gacctcctga ggaagcagcg gacattcgat aacggcagca tcccacacca
     4321 gattcatctc ggggagctgc acgctatcct gaggaggcag gaggacttct accctttcct
     4381 caaggataac cgcgagaaga tcgagaagat tctgactttc aggatcccgt actacgtcgg
     4441 cccactcgct aggggcaact cccgcttcgc ttggatgacc cgcaagtcag aggagacgat
     4501 cacgccgtgg aacttcgagg aggtggtcga caagggcgct agcgctcagt cgttcatcga
     4561 gaggatgacg aatttcgaca agaacctgcc aaatgagaag gtgctcccta agcactcgct
     4621 cctgtacgag tacttcacag tctacaacga gctgactaag gtgaagtatg tgaccgaggg
     4681 catgaggaag ccggctttcc tgtctgggga gcagaagaag gccatcgtgg acctcctgtt
     4741 caagaccaac cggaaggtca cggttaagca gctcaaggag gactacttca agaagattga
     4801 gtgcttcgat tcggtcgaga tctctggcgt tgaggaccgc ttcaacgcct ccctggggac
     4861 ctaccacgat ctcctgaaga tcattaagga taaggacttc ctggacaacg aggagaatga
     4921 ggatatcctc gaggacattg tgctgacact cactctgttc gaggaccggg agatgatcga
     4981 ggagcgcctg aagacttacg cccatctctt cgatgacaag gtcatgaagc agctcaagag
     5041 gaggaggtac accggctggg ggaggctgag caggaagctc atcaacggca ttcgggacaa
     5101 gcagtccggg aagacgatcc tcgacttcct gaagagcgat ggcttcgcga accgcaattt
     5161 catgcagctg attcacgatg acagcctcac attcaaggag gatatccaga aggctcaggt
     5221 gagcggccag ggggactcgc tgcacgagca tatcgcgaac ctcgctggct cgccagctat
     5281 caagaagggg attctgcaga ccgtgaaggt tgtggacgag ctggtgaagg tcatgggcag
     5341 gcacaagcct gagaacatcg tcattgagat ggcccgggag aatcagacca cgcagaaggg
     5401 ccagaagaac tcacgcgaga ggatgaagag gatcgaggag ggcattaagg agctggggtc
     5461 ccagatcctc aaggagcacc cggtggagaa cacgcagctg cagaatgaga agctctacct
     5521 gtactacctc cagaatggcc gcgatatgta tgtggaccag gagctggata ttaacaggct
     5581 cagcgattac gacgtcgatg ccatcgttcc acagtcattc ctgaaggatg actccattga
     5641 caacaaggtc ctcaccaggt cggacaagaa ccggggcaag tctgataatg ttccttcaga
     5701 ggaggtcgtt aagaagatga agaactactg gcgccagctc ctgaatgcca agctgatcac
     5761 gcagcggaag ttcgataacc tcacaaaggc tgagaggggc gggctctctg agctggacaa
     5821 ggcgggcttc atcaagaggc agctggtcga gacacggcag atcactaagc acgttgcgca
     5881 gattctcgac tcacggatga acactaagta cgatgagaat gacaagctga tccgcgaggt
     5941 gaaggtcatc accctgaagt caaagctcgt ctccgacttc aggaaggatt tccagttcta
     6001 caaggttcgg gagatcaaca attaccacca tgcccatgac gcgtacctga acgcggtggt
     6061 cggcacagct ctgatcaaga agtacccaaa gctcgagagc gagttcgtgt acggggacta
     6121 caaggtttac gatgtgagga agatgatcgc caagtcggag caggagattg gcaaggctac
     6181 cgccaagtac ttcttctact ctaacattat gaatttcttc aagacagaga tcactctggc
     6241 caatggcgag atccggaagc gccccctcat cgagacgaac ggcgagacgg gggagatcgt
     6301 gtgggacaag ggcagggatt tcgcgaccgt caggaaggtt ctctccatgc cacaagtgaa
     6361 tatcgtcaag aagacagagg tccagactgg cgggttctct aaggagtcaa ttctgcctaa
     6421 gcggaacagc gacaagctca tcgcccgcaa gaaggactgg gatccgaaga agtacggcgg
     6481 gttcgacagc cccactgtgg cctactcggt cctggttgtg gcgaaggttg agaagggcaa
     6541 gtccaagaag ctcaagagcg tgaaggagct gctggggatc acgattatgg agcgctccag
     6601 cttcgagaag aacccgatcg atttcctgga ggcgaagggc tacaaggagg tgaagaagga
     6661 cctgatcatt aagctcccca agtactcact cttcgagctg gagaacggca ggaagcggat
     6721 gctggcttcc gctggcgagc tgcagaaggg gaacgagctg gctctgccgt ccaagtatgt
     6781 gaacttcctc tacctggcct cccactacga gaagctcaag ggcagccccg aggacaacga
     6841 gcagaagcag ctgttcgtcg agcagcacaa gcattacctc gacgagatca ttgagcagat
     6901 ttccgagttc tccaagcgcg tgatcctggc cgacgcgaat ctggataagg tcctctccgc
     6961 gtacaacaag caccgcgaca agccaatcag ggagcaggct gagaatatca ttcatctctt
     7021 caccctgacg aacctcggcg cccctgctgc tttcaagtac ttcgacacaa ctatcgatcg
     7081 caagaggtac acaagcacta aggaggtcct ggacgcgacc ctcatccacc agtcgattac
     7141 cggcctctac gagacgcgca tcgacctgtc tcagctcggg ggcgacaagc ggccagcggc
     7201 gacgaagaag gcggggcagg cgaagaagaa gaagtccggg tcctcggggt cgccaaagaa
     7261 gaagcggaag gtcggctcaa gcgggtcacc aaagaagaag cggaaggtcg gctcgatgga
     7321 cgctaagtcc ctcaccgctt ggagccgcac cctggtgacg ttcaaggacg tgttcgtcga
     7381 tttcacaagg gaggagtgga agctcctgga cactgctcag cagatcgttt accggaacgt
     7441 gatgctggag aactacaaga acctcgtgtc cctcggctac cagctgacga agccggatgt
     7501 tattctccgc ctggagaagg gcgaggagcc cgggtcccct aagaagaagc ggaaggtcgg
     7561 ctcctaccca tacgatgtcc ctgactacgc ctcatgagct caggcctcca tacaagtatt
     7621 ggggatccac tacgaatttc cccgatcgtt caaacatttg gcaataaagt ttcttaagat
     7681 tgaatcctgt tgccggtctt gcgatgatta tcatataatt tctgttgaat tacgttaagc
     7741 atgtaataat taacatgtaa tgcatgacgt tatttatgag atgggttttt atgattagag
     7801 tcccgcaatt atacatttaa tacgcgatag aaaacaaaat atagcgcgca aactaggata
     7861 aattatcgcg cgcggtgtca tctatgttac tagatcggaa ttcgaggcgg tttgcgtatt
     7921 ggctagagca gcttgccaac atggtggagc acgacactct cgtctactcc aagaatatca
     7981 aagatacagt ctcagaagac caaagggcta ttgagacttt tcaacaaagg gtaatatcgg
     8041 gaaacctcct cggattccat tgcccagcta tctgtcactt catcaaaagg acagtagaaa
     8101 aggaaggtgg cacctacaaa tgccatcatt gcgataaagg aaaggctatc gttcaagatg
     8161 cctctgccga cagtggtccc aaagatggac ccccacccac gaggagcatc gtggaaaaag
     8221 aagacgttcc aaccacgtct tcaaagcaag tggattgatg tgataacatg gtggagcacg
     8281 acactctcgt ctactccaag aatatcaaag atacagtctc agaagaccaa agggctattg
     8341 agacttttca acaaagggta atatcgggaa acctcctcgg attccattgc ccagctatct
     8401 gtcacttcat caaaaggaca gtagaaaagg aaggtggcac ctacaaatgc catcattgcg
     8461 ataaaggaaa ggctatcgtt caagatgcct ctgccgacag tggtcccaaa gatggacccc
     8521 cacccacgag gagcatcgtg gaaaaagaag acgttccaac cacgtcttca aagcaagtgg
     8581 attgatgtga tatctccact gacgtaaggg atgacgcaca atcccactat ccttcgcaag
     8641 accttcctct atataaggaa gttcatttca tttggagagg acacgctgaa atcaccagtc
     8701 tctctctaca aatctatctc tctcgagctt tcgcagatcc cggggggcaa tgagatatga
     8761 aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg
     8821 tctccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag
     8881 gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt
     8941 atgtttatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt gacattgggg
     9001 agtttagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag
     9061 acctgcctga aaccgaactg cccgctgttc tacaaccggt cgcggaggct atggatgcga
     9121 tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg
     9181 gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact
     9241 ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga
     9301 tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca
     9361 acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt
     9421 tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta
     9481 tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccacgac
     9541 tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca
     9601 atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg
     9661 ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc gatggctgtg
     9721 tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg gcaaagaaat
     9781 agagtagatg ccgaccggat ctgtcgatcg acaagctcga gtttctccat aataatgtgt
     9841 gagtagttcc cagataaggg aattagggtt cctatagggt ttcgctcatg tgttgagcat
     9901 ataagaaacc cttagtatgt atttgtattt gtaaaatact tctatcaata aaatttctaa
     9961 ttcctaaaac caaaatccag tactaaaatc cagatccccc gaattaattc ggcgttaatt
    10021 cagtacatta aaaacgtccg caatgtgtta ttaagttgtc actagtcagg ttaactcaat
    10081 tcggcgttaa ttcagtacat taaaaacgtc cgcaatgtgt tattaagttg tctaagcgtc
    10141 aatttgttta caccacaata tatcctgcca ccagccagcc aacagctccc cgaccggcag
    10201 ctcggcacaa aatcaccact cgatacaggc agcccatcag tccgggacgg cgtcagcggg
    10261 agagccgttg taaggcggca gactttgctc atgttaccga tgctattcgg aagaacggca
    10321 actaagctgc cgggtttgaa acacggatga tctcgcggag ggtagcatgt tgattgtaac
    10381 gatgacagag cgttgctgcc tgtgatcact taagtaacta actaacagga agagtttgta
    10441 gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt agtttgatgc ctggcagttt
    10501 atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc aaatccgctc
    10561 ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg
    10621 cccagtcttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgct
    10681 tagtagttag acgtccccga gatccatgct agaccatgaa tccagaagcc cgagaggttg
    10741 ccgcctttcg ggctttttct ttttcaaaaa aaaaaattta taaaacgatc tgttgcggcc
    10801 ggccgccggg ttgtgggcaa aggcgctcga cggtgggcaa ccgcttgcgg ttgtccacgg
    10861 gcggagccgg tgcgcgtagc gcattgtcca caagccaagg gcgaccaata attgatatat
    10921 atattcataa ttgaaaagct aattgaacat actacttgct gtaactactt gccggagcga
    10981 ggggtgtttg caagctgttg atctgaaagg gctattagcg ttctcacgtg cctttttgat
    11041 tagcgatttc acgtgacctt attagcgatt tcacgtactc cgattagcga tttcacgtac
    11101 cctgattagc gatttcacgt ggatagtttt tggagcgggc cggaaagccc cgtgaatcaa
    11161 ggctttgcgg ggcattagcg gtttcacgtg gataactacc ctctatccac aggcttccgg
    11221 ggataaaaaa gcccgctcga cggcgggctg ttggatgggg atctagcggt aatacggtta
    11281 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
    11341 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
    11401 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
    11461 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
    11521 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
    11581 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
    11641 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
    11701 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
    11761 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
    11821 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
    11881 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
    11941 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctaccgggtc tgacgctcag
    12001 tggaacgggg cccaatctga ataatgttac aaccaattaa ccaattctga ttagaaaaac
    12061 tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt
    12121 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca
    12181 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc
    12241 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt
    12301 gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc
    12361 tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg
    12421 agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg
    12481 cgcagggaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat
    12541 acctggaatg ctgtttttcc ggggatcgca gtggtgagta accatgcatc atcaggagta
    12601 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc
    12661 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc
    12721 gcatcgggct tcccatacaa gcgatagatt gtcgcacctg attgcccgac attatcgcga
    12781 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg cctcgacgtt
    12841 tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt
    12901 attgttcatg atgatatatt tttatcttgt gcaatgtaac atcagagatt ttgagacacg
    12961 ggccagagct gcagtttgat cccgagggga accctgtggt tgacatgcac atacaaatgg
    13021 acgaacggat aaaccttttc acgccctttt aaatatccgt tattctaata aacgctcttt
    13081 tctcttag
//