LOCUS       sequence-116214-       10577 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10577)
  AUTHORS   Jinek M, Chylinski K, Fonfara I, Hauer M, Doudna JA, Charpentier E
  TITLE     A Programmable Dual-RNA-Guided DNA Endonuclease in Adaptive 
            Bacterial Immunity.
  JOURNAL   Science. 2012 Aug 17;337(6096):816-21. doi: 10.1126/science.1225829.
            Epub 2012 Jun 28.
  PUBMED    22745249
REFERENCE   2  (bases 1 to 10577)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1126/science.1225829"; journalName: "Science"; date: 
            "2012-08-17- 17"; volume: "337"; issue: "6096"; pages: "816-21"
COMMENT     Sequence Label: pMJ806
FEATURES             Location/Qualifiers
     source          1..10577
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    5..144
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(70..89)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     230..252
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(246..437)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1246..2328)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     2290..2309
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2329..2406)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2612..2631
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2715..2734
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2715..2733
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2734..2758
                     /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)."
     RBS             2773..2795
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2806..2823
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2827..3924
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="KIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
                     KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
                     AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
                     DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
                     AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
                     NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
                     YAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     3898..3921
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             3955..3975
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3985..8091
                     /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             3985..8088
                     /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             8156..8173
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(8226..8244)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      8240..8287
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      8324..8779
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8411..8430)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8621..8642
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(8871..9686)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(8871..9686)
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     9594..9613
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     9678..9697
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      9808..10396
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10297..10316
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10550..10567
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg
       61 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
      121 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
      181 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
      241 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca
      301 tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg
      361 agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt
      421 ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg
      481 ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg
      541 ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa
      601 atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc
      661 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt
      721 tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac
      781 gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca
      841 gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc
      901 cgtggggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga
      961 ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg
     1021 atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc
     1081 acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg
     1141 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat
     1201 cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc
     1261 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     1321 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
     1381 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
     1441 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
     1501 ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat
     1561 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
     1621 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
     1681 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
     1741 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
     1801 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
     1861 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
     1921 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
     1981 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
     2041 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
     2101 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
     2161 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
     2221 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
     2281 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
     2341 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat
     2401 ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta
     2461 gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc
     2521 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
     2581 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
     2641 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatct
     2701 cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata acaattcccc
     2761 tctagaaata attttgttta actttaagaa ggagatatac atatgcacca tcaccatcac
     2821 catggaaaaa tcgaagaagg taaactggta atctggatta acggcgataa aggctataac
     2881 ggtctcgctg aagtcggtaa gaaattcgag aaagataccg gaattaaagt caccgttgag
     2941 catccggata aactggaaga gaaattccca caggttgcgg caactggcga tggccctgac
     3001 attatcttct gggcacacga ccgctttggt ggctacgctc aatctggcct gttggctgaa
     3061 atcaccccgg acaaagcgtt ccaggacaag ctgtatccgt ttacctggga tgccgtacgt
     3121 tacaacggca agctgattgc ttacccgatc gctgttgaag cgttatcgct gatttataac
     3181 aaagatctgc tgccgaaccc gccaaaaacc tgggaagaga tcccggcgct ggataaagaa
     3241 ctgaaagcga aaggtaagag cgcgctgatg ttcaacctgc aagaaccgta cttcacctgg
     3301 ccgctgattg ctgctgacgg gggttatgcg ttcaagtatg aaaacggcaa gtacgacatt
     3361 aaagacgtgg gcgtggataa cgctggcgcg aaagcgggtc tgaccttcct ggttgacctg
     3421 attaaaaaca aacacatgaa tgcagacacc gattactcca tcgcagaagc tgcctttaat
     3481 aaaggcgaaa cagcgatgac catcaacggc ccgtgggcat ggtccaacat cgacaccagc
     3541 aaagtgaatt atggtgtaac ggtactgccg accttcaagg gtcaaccatc caaaccgttc
     3601 gttggcgtgc tgagcgcagg tattaacgcc gccagtccga acaaagagct ggcaaaagag
     3661 ttcctcgaaa actatctgct gactgatgaa ggtctggaag cggttaataa agacaaaccg
     3721 ctgggtgccg tagcgctgaa gtcttacgag gaagagttgg cgaaagatcc acgtattgcc
     3781 gccactatgg aaaacgccca gaaaggtgaa atcatgccga acatcccgca gatgtccgct
     3841 ttctggtatg ccgtgcgtac tgcggtgatc aacgccgcca gcggtcgtca gactgtcgat
     3901 gaagccctga aagacgcgca gactaattcg agctcgaaca acaacaacac tagtgaaaac
     3961 ctgtatttcc agggagcagc ctcgatggat aagaaatact caataggctt agatatcggc
     4021 acaaatagcg tcggatgggc ggtgatcact gatgaatata aggttccgtc taaaaagttc
     4081 aaggttctgg gaaatacaga ccgccacagt atcaaaaaaa atcttatagg ggctctttta
     4141 tttgacagtg gagagacagc ggaagcgact cgtctcaaac ggacagctcg tagaaggtat
     4201 acacgtcgga agaatcgtat ttgttatcta caggagattt tttcaaatga gatggcgaaa
     4261 gtagatgata gtttctttca tcgacttgaa gagtcttttt tggtggaaga agacaagaag
     4321 catgaacgtc atcctatttt tggaaatata gtagatgaag ttgcttatca tgagaaatat
     4381 ccaactatct atcatctgcg aaaaaaattg gtagattcta ctgataaagc ggatttgcgc
     4441 ttaatctatt tggccttagc gcatatgatt aagtttcgtg gtcatttttt gattgaggga
     4501 gatttaaatc ctgataatag tgatgtggac aaactattta tccagttggt acaaacctac
     4561 aatcaattat ttgaagaaaa ccctattaac gcaagtggag tagatgctaa agcgattctt
     4621 tctgcacgat tgagtaaatc aagacgatta gaaaatctca ttgctcagct ccccggtgag
     4681 aagaaaaatg gcttatttgg gaatctcatt gctttgtcat tgggtttgac ccctaatttt
     4741 aaatcaaatt ttgatttggc agaagatgct aaattacagc tttcaaaaga tacttacgat
     4801 gatgatttag ataatttatt ggcgcaaatt ggagatcaat atgctgattt gtttttggca
     4861 gctaagaatt tatcagatgc tattttactt tcagatatcc taagagtaaa tactgaaata
     4921 actaaggctc ccctatcagc ttcaatgatt aaacgctacg atgaacatca tcaagacttg
     4981 actcttttaa aagctttagt tcgacaacaa cttccagaaa agtataaaga aatctttttt
     5041 gatcaatcaa aaaacggata tgcaggttat attgatgggg gagctagcca agaagaattt
     5101 tataaattta tcaaaccaat tttagaaaaa atggatggta ctgaggaatt attggtgaaa
     5161 ctaaatcgtg aagatttgct gcgcaagcaa cggacctttg acaacggctc tattccccat
     5221 caaattcact tgggtgagct gcatgctatt ttgagaagac aagaagactt ttatccattt
     5281 ttaaaagaca atcgtgagaa gattgaaaaa atcttgactt ttcgaattcc ttattatgtt
     5341 ggtccattgg cgcgtggcaa tagtcgtttt gcatggatga ctcggaagtc tgaagaaaca
     5401 attaccccat ggaattttga agaagttgtc gataaaggtg cttcagctca atcatttatt
     5461 gaacgcatga caaactttga taaaaatctt ccaaatgaaa aagtactacc aaaacatagt
     5521 ttgctttatg agtattttac ggtttataac gaattgacaa aggtcaaata tgttactgaa
     5581 ggaatgcgaa aaccagcatt tctttcaggt gaacagaaga aagccattgt tgatttactc
     5641 ttcaaaacaa atcgaaaagt aaccgttaag caattaaaag aagattattt caaaaaaata
     5701 gaatgttttg atagtgttga aatttcagga gttgaagata gatttaatgc ttcattaggt
     5761 acctaccatg atttgctaaa aattattaaa gataaagatt ttttggataa tgaagaaaat
     5821 gaagatatct tagaggatat tgttttaaca ttgaccttat ttgaagatag ggagatgatt
     5881 gaggaaagac ttaaaacata tgctcacctc tttgatgata aggtgatgaa acagcttaaa
     5941 cgtcgccgtt atactggttg gggacgtttg tctcgaaaat tgattaatgg tattagggat
     6001 aagcaatctg gcaaaacaat attagatttt ttgaaatcag atggttttgc caatcgcaat
     6061 tttatgcagc tgatccatga tgatagtttg acatttaaag aagacattca aaaagcacaa
     6121 gtgtctggac aaggcgatag tttacatgaa catattgcaa atttagctgg tagccctgct
     6181 attaaaaaag gtattttaca gactgtaaaa gttgttgatg aattggtcaa agtaatgggg
     6241 cggcataagc cagaaaatat cgttattgaa atggcacgtg aaaatcagac aactcaaaag
     6301 ggccagaaaa attcgcgaga gcgtatgaaa cgaatcgaag aaggtatcaa agaattagga
     6361 agtcagattc ttaaagagca tcctgttgaa aatactcaat tgcaaaatga aaagctctat
     6421 ctctattatc tccaaaatgg aagagacatg tatgtggacc aagaattaga tattaatcgt
     6481 ttaagtgatt atgatgtcga tcacattgtt ccacaaagtt tccttaaaga cgattcaata
     6541 gacaataagg tcttaacgcg ttctgataaa aatcgtggta aatcggataa cgttccaagt
     6601 gaagaagtag tcaaaaagat gaaaaactat tggagacaac ttctaaacgc caagttaatc
     6661 actcaacgta agtttgataa tttaacgaaa gctgaacgtg gaggtttgag tgaacttgat
     6721 aaagctggtt ttatcaaacg ccaattggtt gaaactcgcc aaatcactaa gcatgtggca
     6781 caaattttgg atagtcgcat gaatactaaa tacgatgaaa atgataaact tattcgagag
     6841 gttaaagtga ttaccttaaa atctaaatta gtttctgact tccgaaaaga tttccaattc
     6901 tataaagtac gtgagattaa caattaccat catgcccatg atgcgtatct aaatgccgtc
     6961 gttggaactg ctttgattaa gaaatatcca aaacttgaat cggagtttgt ctatggtgat
     7021 tataaagttt atgatgttcg taaaatgatt gctaagtctg agcaagaaat aggcaaagca
     7081 accgcaaaat atttctttta ctctaatatc atgaacttct tcaaaacaga aattacactt
     7141 gcaaatggag agattcgcaa acgccctcta atcgaaacta atggggaaac tggagaaatt
     7201 gtctgggata aagggcgaga ttttgccaca gtgcgcaaag tattgtccat gccccaagtc
     7261 aatattgtca agaaaacaga agtacagaca ggcggattct ccaaggagtc aattttacca
     7321 aaaagaaatt cggacaagct tattgctcgt aaaaaagact gggatccaaa aaaatatggt
     7381 ggttttgata gtccaacggt agcttattca gtcctagtgg ttgctaaggt ggaaaaaggg
     7441 aaatcgaaga agttaaaatc cgttaaagag ttactaggga tcacaattat ggaaagaagt
     7501 tcctttgaaa aaaatccgat tgacttttta gaagctaaag gatataagga agttaaaaaa
     7561 gacttaatca ttaaactacc taaatatagt ctttttgagt tagaaaacgg tcgtaaacgg
     7621 atgctggcta gtgccggaga attacaaaaa ggaaatgagc tggctctgcc aagcaaatat
     7681 gtgaattttt tatatttagc tagtcattat gaaaagttga agggtagtcc agaagataac
     7741 gaacaaaaac aattgtttgt ggagcagcat aagcattatt tagatgagat tattgagcaa
     7801 atcagtgaat tttctaagcg tgttatttta gcagatgcca atttagataa agttcttagt
     7861 gcatataaca aacatagaga caaaccaata cgtgaacaag cagaaaatat tattcattta
     7921 tttacgttga cgaatcttgg agctcccgct gcttttaaat attttgatac aacaattgat
     7981 cgtaaacgat atacgtctac aaaagaagtt ttagatgcca ctcttatcca tcaatccatc
     8041 actggtcttt atgaaacacg cattgatttg agtcagctag gaggtgacta acgaggccgg
     8101 tgctttgcag gatccgaatt cgagctccgt cgacaagctt gcggccgcac tcgagcacca
     8161 ccaccaccac cactgagatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     8221 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     8281 ttttttgctg aaaggaggaa ctatatccgg attggcgaat gggacgcgcc ctgtagcggc
     8341 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     8401 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     8461 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     8521 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     8581 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     8641 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     8701 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     8761 atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     8821 gtttattttt ctaaatacat tcaaatatgt atccgctcat gaattaattc ttagaaaaac
     8881 tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt
     8941 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca
     9001 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc
     9061 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt
     9121 gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc
     9181 tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg
     9241 agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg
     9301 cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat
     9361 acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta
     9421 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc
     9481 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc
     9541 gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga
     9601 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg cctagagcaa
     9661 gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac
     9721 agttttattg ttcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     9781 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     9841 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     9901 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     9961 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
    10021 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
    10081 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
    10141 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
    10201 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
    10261 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
    10321 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
    10381 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
    10441 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
    10501 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
    10561 gagcgaggaa gcggaag
//
