LOCUS       sequence-198599-        9262 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Cas9-NLS-6xHis in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9262)
  AUTHORS   Zuris JA, Thompson DB, Shu Y, Guilinger JP, Bessen JL, Hu JH, Maeder
            ML, Joung JK, Chen ZY, Liu DR
  TITLE     Cationic lipid-mediated delivery of proteins enables efficient 
            protein-based genome editing in vitro and in vivo.
  JOURNAL   Nat Biotechnol. 2015 Jan;33(1):73-80. doi: 10.1038/nbt.3081. Epub 
            2014 Oct 30.
  PUBMED    25357182
REFERENCE   2  (bases 1 to 9262)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1038/nbt.3081"; 
            journalName: "Nat Biotechnol"; date: "2015-01"; volume: "33"; issue:
            "1"; pages: "73-80"
COMMENT     Sequence Label: pET-Cas9-NLS-6xHis
FEATURES             Location/Qualifiers
     source          1..9262
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             88..4191
                     /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             88..4191
                     /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             4192..4212
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4222..4239
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4310..4328)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4324..4371
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4408..4863
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4495..4514)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4705..4726
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(4978..5769,5770..5838))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5601..5620
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5839..5931)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6012..6600
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6501..6520
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6754..6771
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6786..6925
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6851..6870)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7011..7033
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(7027..7218)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(7790..8872)
                     /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     8834..8853
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(8873..8950)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     9156..9175
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg gataagaaat actcaatagg cttagatatc
      121 ggcacaaata gcgtcggatg ggcggtgatc actgatgaat ataaggttcc gtctaaaaag
      181 ttcaaggttc tgggaaatac agaccgccac agtatcaaaa aaaatcttat aggggctctt
      241 ttatttgaca gtggagagac agcggaagcg actcgtctca aacggacagc tcgtagaagg
      301 tatacacgtc ggaagaatcg tatttgttat ctacaggaga ttttttcaaa tgagatggcg
      361 aaagtagatg atagtttctt tcatcgactt gaagagtctt ttttggtgga agaagacaag
      421 aagcatgaac gtcatcctat ttttggaaat atagtagatg aagttgctta tcatgagaaa
      481 tatccaacta tctatcatct gcgaaaaaaa ttggtagatt ctactgataa agcggatttg
      541 cgcttaatct atttggcctt agcgcatatg attaagtttc gtggtcattt tttgattgag
      601 ggagatttaa atcctgataa tagtgatgtg gacaaactat ttatccagtt ggtacaaacc
      661 tacaatcaat tatttgaaga aaaccctatt aacgcaagtg gagtagatgc taaagcgatt
      721 ctttctgcac gattgagtaa atcaagacga ttagaaaatc tcattgctca gctccccggt
      781 gagaagaaaa atggcttatt tgggaatctc attgctttgt cattgggttt gacccctaat
      841 tttaaatcaa attttgattt ggcagaagat gctaaattac agctttcaaa agatacttac
      901 gatgatgatt tagataattt attggcgcaa attggagatc aatatgctga tttgtttttg
      961 gcagctaaga atttatcaga tgctatttta ctttcagata tcctaagagt aaatactgaa
     1021 ataactaagg ctcccctatc agcttcaatg attaaacgct acgatgaaca tcatcaagac
     1081 ttgactcttt taaaagcttt agttcgacaa caacttccag aaaagtataa agaaatcttt
     1141 tttgatcaat caaaaaacgg atatgcaggt tatattgatg ggggagctag ccaagaagaa
     1201 ttttataaat ttatcaaacc aattttagaa aaaatggatg gtactgagga attattggtg
     1261 aaactaaatc gtgaagattt gctgcgcaag caacggacct ttgacaacgg ctctattccc
     1321 catcaaattc acttgggtga gctgcatgct attttgagaa gacaagaaga cttttatcca
     1381 tttttaaaag acaatcgtga gaagattgaa aaaatcttga cttttcgaat tccttattat
     1441 gttggtccat tggcgcgtgg caatagtcgt tttgcatgga tgactcggaa gtctgaagaa
     1501 acaattaccc catggaattt tgaagaagtt gtcgataaag gtgcttcagc tcaatcattt
     1561 attgaacgca tgacaaactt tgataaaaat cttccaaatg aaaaagtact accaaaacat
     1621 agtttgcttt atgagtattt tacggtttat aacgaattga caaaggtcaa atatgttact
     1681 gaaggaatgc gaaaaccagc atttctttca ggtgaacaga agaaagccat tgttgattta
     1741 ctcttcaaaa caaatcgaaa agtaaccgtt aagcaattaa aagaagatta tttcaaaaaa
     1801 atagaatgtt ttgatagtgt tgaaatttca ggagttgaag atagatttaa tgcttcatta
     1861 ggtacctacc atgatttgct aaaaattatt aaagataaag attttttgga taatgaagaa
     1921 aatgaagata tcttagagga tattgtttta acattgacct tatttgaaga tagggagatg
     1981 attgaggaaa gacttaaaac atatgctcac ctctttgatg ataaggtgat gaaacagctt
     2041 aaacgtcgcc gttatactgg ttggggacgt ttgtctcgaa aattgattaa tggtattagg
     2101 gataagcaat ctggcaaaac aatattagat tttttgaaat cagatggttt tgccaatcgc
     2161 aattttatgc agctgatcca tgatgatagt ttgacattta aagaagacat tcaaaaagca
     2221 caagtgtctg gacaaggcga tagtttacat gaacatattg caaatttagc tggtagccct
     2281 gctattaaaa aaggtatttt acagactgta aaagttgttg atgaattggt caaagtaatg
     2341 gggcggcata agccagaaaa tatcgttatt gaaatggcac gtgaaaatca gacaactcaa
     2401 aagggccaga aaaattcgcg agagcgtatg aaacgaatcg aagaaggtat caaagaatta
     2461 ggaagtcaga ttcttaaaga gcatcctgtt gaaaatactc aattgcaaaa tgaaaagctc
     2521 tatctctatt atctccaaaa tggaagagac atgtatgtgg accaagaatt agatattaat
     2581 cgtttaagtg attatgatgt cgatcacatt gttccacaaa gtttccttaa agacgattca
     2641 atagacaata aggtcttaac gcgttctgat aaaaatcgtg gtaaatcgga taacgttcca
     2701 agtgaagaag tagtcaaaaa gatgaaaaac tattggagac aacttctaaa cgccaagtta
     2761 atcactcaac gtaagtttga taatttaacg aaagctgaac gtggaggttt gagtgaactt
     2821 gataaagctg gttttatcaa acgccaattg gttgaaactc gccaaatcac taagcatgtg
     2881 gcacaaattt tggatagtcg catgaatact aaatacgatg aaaatgataa acttattcga
     2941 gaggttaaag tgattacctt aaaatctaaa ttagtttctg acttccgaaa agatttccaa
     3001 ttctataaag tacgtgagat taacaattac catcatgccc atgatgcgta tctaaatgcc
     3061 gtcgttggaa ctgctttgat taagaaatat ccaaaacttg aatcggagtt tgtctatggt
     3121 gattataaag tttatgatgt tcgtaaaatg attgctaagt ctgagcaaga aataggcaaa
     3181 gcaaccgcaa aatatttctt ttactctaat atcatgaact tcttcaaaac agaaattaca
     3241 cttgcaaatg gagagattcg caaacgccct ctaatcgaaa ctaatgggga aactggagaa
     3301 attgtctggg ataaagggcg agattttgcc acagtgcgca aagtattgtc catgccccaa
     3361 gtcaatattg tcaagaaaac agaagtacag acaggcggat tctccaagga gtcaatttta
     3421 ccaaaaagaa attcggacaa gcttattgct cgtaaaaaag actgggatcc aaaaaaatat
     3481 ggtggttttg atagtccaac ggtagcttat tcagtcctag tggttgctaa ggtggaaaaa
     3541 gggaaatcga agaagttaaa atccgttaaa gagttactag ggatcacaat tatggaaaga
     3601 agttcctttg aaaaaaatcc gattgacttt ttagaagcta aaggatataa ggaagttaaa
     3661 aaagacttaa tcattaaact acctaaatat agtctttttg agttagaaaa cggtcgtaaa
     3721 cggatgctgg ctagtgccgg agaattacaa aaaggaaatg agctggctct gccaagcaaa
     3781 tatgtgaatt ttttatattt agctagtcat tatgaaaagt tgaagggtag tccagaagat
     3841 aacgaacaaa aacaattgtt tgtggagcag cataagcatt atttagatga gattattgag
     3901 caaatcagtg aattttctaa gcgtgttatt ttagcagatg ccaatttaga taaagttctt
     3961 agtgcatata acaaacatag agacaaacca atacgtgaac aagcagaaaa tattattcat
     4021 ttatttacgt tgacgaatct tggagctccc gctgctttta aatattttga tacaacaatt
     4081 gatcgtaaac gatatacgtc tacaaaagaa gttttagatg ccactcttat ccatcaatcc
     4141 atcactggtc tttatgaaac acgcattgat ttgagtcagc taggaggtga ccccaagaag
     4201 aagaggaagg tgatggataa gcatcaccac caccatcact aatgataatt tgaacgccag
     4261 cacatggact cgtctactag cgcagcttaa ttaacctagg ctgctgccac cgctgagcaa
     4321 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga
     4381 ggaactatat ccggattggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
     4441 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     4501 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     4561 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     4621 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     4681 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     4741 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     4801 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     4861 tttctggcgg cacgatggca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4921 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4981 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5041 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5101 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5161 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5221 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5281 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5341 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5401 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5461 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5521 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5581 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5641 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5701 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5761 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5821 gaaatgttga atactcatac tcttcctttt tcaatcatga ttgaagcatt tatcagggtt
     5881 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggtcatg
     5941 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     6001 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     6061 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     6121 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     6181 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     6241 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     6301 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     6361 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     6421 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     6481 cgcacgaggg agctcccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     6541 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     6601 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     6661 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     6721 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     6781 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt
     6841 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc
     6901 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg
     6961 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     7021 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc
     7081 atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt
     7141 gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt
     7201 tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat
     7261 gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg
     7321 gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa
     7381 aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag
     7441 ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt
     7501 ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga
     7561 cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc
     7621 agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgctagt
     7681 catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg
     7741 agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc
     7801 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     7861 aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca
     7921 gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt
     7981 gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt
     8041 cttcggtatc gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc ccggactcgg
     8101 taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa
     8161 cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc
     8221 cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca
     8281 gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac
     8341 ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac
     8401 tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag
     8461 cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc
     8521 gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca
     8581 tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt
     8641 gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc
     8701 ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca
     8761 ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct
     8821 gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca
     8881 ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt
     8941 cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag gaagcagccc
     9001 agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg
     9061 gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc
     9121 atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca
     9181 gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcgag
     9241 atcgatctcg atcccgcgaa at
//
