LOCUS       sequence-120935-        9840 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cas9nlsHis6 Bacterial Expression construct.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9840)
  AUTHORS   Fu BX, Hansen LL, Artiles KL, Nonet ML, Fire AZ
  TITLE     Landscape of target:guide homology effects on Cas9-mediated 
            cleavage.
  JOURNAL   Nucleic Acids Res. 2014 Dec 16;42(22):13778-87. doi: 
            10.1093/nar/gku1102. Epub 2014 Nov 15.
  PUBMED    25399416
REFERENCE   2  (bases 1 to 9840)
  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: 
            "10.1093/nar/gku1102"; journalName: "Nucleic Acids Res"; date: 
            "2014-12-16- 16"; volume: "42"; issue: "22"; pages: "13778-87"
COMMENT     Sequence Label: pHO4d-Cas9
FEATURES             Location/Qualifiers
     source          1..9840
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             13..35
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             42..4145
                     /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             42..4145
                     /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             4152..4172
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4203..4220
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4224..4253
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(4309..4327)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4323..4370
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4582..4600)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4668..4772
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4773..4841,4842..5633)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4991..5010)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5804..6392
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6293..6312
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6546..6563
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6578..6720
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6646..6665)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6806..6828
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     protein_bind    8292..8313
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(8326..9408)
                     /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
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTANQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     9370..9389
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(9409..9486)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     9692..9711
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     9795..9814
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        9795..9813
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    9814..9838
                     /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)."
ORIGIN
        1 tctagaaata attttgttta actttaagaa ggagatatac catggataag aaatactcaa
       61 taggcttaga tatcggcaca aatagcgtcg gatgggcggt gatcactgat gaatataagg
      121 ttccgtctaa aaagttcaag gttctgggaa atacagaccg ccacagtatc aaaaaaaatc
      181 ttataggggc tcttttattt gacagtggag agacagcgga agcgactcgt ctcaaacgga
      241 cagctcgtag aaggtataca cgtcggaaga atcgtatttg ttatctacag gagatttttt
      301 caaatgagat ggcgaaagta gatgatagtt tctttcatcg acttgaagag tcttttttgg
      361 tggaagaaga caagaagcat gaacgtcatc ctatttttgg aaatatagta gatgaagttg
      421 cttatcatga gaaatatcca actatctatc atctgcgaaa aaaattggta gattctactg
      481 ataaagcgga tttgcgctta atctatttgg ccttagcgca tatgattaag tttcgtggtc
      541 attttttgat tgagggagat ttaaatcctg ataatagtga tgtggacaaa ctatttatcc
      601 agttggtaca aacctacaat caattatttg aagaaaaccc tattaacgca agtggagtag
      661 atgctaaagc gattctttct gcacgattga gtaaatcaag acgattagaa aatctcattg
      721 ctcagctccc cggtgagaag aaaaatggct tatttgggaa tctcattgct ttgtcattgg
      781 gtttgacccc taattttaaa tcaaattttg atttggcaga agatgctaaa ttacagcttt
      841 caaaagatac ttacgatgat gatttagata atttattggc gcaaattgga gatcaatatg
      901 ctgatttgtt tttggcagct aagaatttat cagatgctat tttactttca gatatcctaa
      961 gagtaaatac tgaaataact aaggctcccc tatcagcttc aatgattaaa cgctacgatg
     1021 aacatcatca agacttgact cttttaaaag ctttagttcg acaacaactt ccagaaaagt
     1081 ataaagaaat cttttttgat caatcaaaaa acggatatgc aggttatatt gatgggggag
     1141 ctagccaaga agaattttat aaatttatca aaccaatttt agaaaaaatg gatggtactg
     1201 aggaattatt ggtgaaacta aatcgtgaag atttgctgcg caagcaacgg acctttgaca
     1261 acggctctat tccccatcaa attcacttgg gtgagctgca tgctattttg agaagacaag
     1321 aagactttta tccattttta aaagacaatc gtgagaagat tgaaaaaatc ttgacttttc
     1381 gaattcctta ttatgttggt ccattggcgc gtggcaatag tcgttttgca tggatgactc
     1441 ggaagtctga agaaacaatt accccatgga attttgaaga agttgtcgat aaaggtgctt
     1501 cagctcaatc atttattgaa cgcatgacaa actttgataa aaatcttcca aatgaaaaag
     1561 tactaccaaa acatagtttg ctttatgagt attttacggt ttataacgaa ttgacaaagg
     1621 tcaaatatgt tactgaagga atgcgaaaac cagcatttct ttcaggtgaa cagaagaaag
     1681 ccattgttga tttactcttc aaaacaaatc gaaaagtaac cgttaagcaa ttaaaagaag
     1741 attatttcaa aaaaatagaa tgttttgata gtgttgaaat ttcaggagtt gaagatagat
     1801 ttaatgcttc attaggtacc taccatgatt tgctaaaaat tattaaagat aaagattttt
     1861 tggataatga agaaaatgaa gatatcttag aggatattgt tttaacattg accttatttg
     1921 aagataggga gatgattgag gaaagactta aaacatatgc tcacctcttt gatgataagg
     1981 tgatgaaaca gcttaaacgt cgccgttata ctggttgggg acgtttgtct cgaaaattga
     2041 ttaatggtat tagggataag caatctggca aaacaatatt agattttttg aaatcagatg
     2101 gttttgccaa tcgcaatttt atgcagctga tccatgatga tagtttgaca tttaaagaag
     2161 acattcaaaa agcacaagtg tctggacaag gcgatagttt acatgaacat attgcaaatt
     2221 tagctggtag ccctgctatt aaaaaaggta ttttacagac tgtaaaagtt gttgatgaat
     2281 tggtcaaagt aatggggcgg cataagccag aaaatatcgt tattgaaatg gcacgtgaaa
     2341 atcagacaac tcaaaagggc cagaaaaatt cgcgagagcg tatgaaacga atcgaagaag
     2401 gtatcaaaga attaggaagt cagattctta aagagcatcc tgttgaaaat actcaattgc
     2461 aaaatgaaaa gctctatctc tattatctcc aaaatggaag agacatgtat gtggaccaag
     2521 aattagatat taatcgttta agtgattatg atgtcgatca cattgttcca caaagtttcc
     2581 ttaaagacga ttcaatagac aataaggtct taacgcgttc tgataaaaat cgtggtaaat
     2641 cggataacgt tccaagtgaa gaagtagtca aaaagatgaa aaactattgg agacaacttc
     2701 taaacgccaa gttaatcact caacgtaagt ttgataattt aacgaaagct gaacgtggag
     2761 gtttgagtga acttgataaa gctggtttta tcaaacgcca attggttgaa actcgccaaa
     2821 tcactaagca tgtggcacaa attttggata gtcgcatgaa tactaaatac gatgaaaatg
     2881 ataaacttat tcgagaggtt aaagtgatta ccttaaaatc taaattagtt tctgacttcc
     2941 gaaaagattt ccaattctat aaagtacgtg agattaacaa ttaccatcat gcccatgatg
     3001 cgtatctaaa tgccgtcgtt ggaactgctt tgattaagaa atatccaaaa cttgaatcgg
     3061 agtttgtcta tggtgattat aaagtttatg atgttcgtaa aatgattgct aagtctgagc
     3121 aagaaatagg caaagcaacc gcaaaatatt tcttttactc taatatcatg aacttcttca
     3181 aaacagaaat tacacttgca aatggagaga ttcgcaaacg ccctctaatc gaaactaatg
     3241 gggaaactgg agaaattgtc tgggataaag ggcgagattt tgccacagtg cgcaaagtat
     3301 tgtccatgcc ccaagtcaat attgtcaaga aaacagaagt acagacaggc ggattctcca
     3361 aggagtcaat tttaccaaaa agaaattcgg acaagcttat tgctcgtaaa aaagactggg
     3421 atccaaaaaa atatggtggt tttgatagtc caacggtagc ttattcagtc ctagtggttg
     3481 ctaaggtgga aaaagggaaa tcgaagaagt taaaatccgt taaagagtta ctagggatca
     3541 caattatgga aagaagttcc tttgaaaaaa atccgattga ctttttagaa gctaaaggat
     3601 ataaggaagt taaaaaagac ttaatcatta aactacctaa atatagtctt tttgagttag
     3661 aaaacggtcg taaacggatg ctggctagtg ccggagaatt acaaaaagga aatgagctgg
     3721 ctctgccaag caaatatgtg aattttttat atttagctag tcattatgaa aagttgaagg
     3781 gtagtccaga agataacgaa caaaaacaat tgtttgtgga gcagcataag cattatttag
     3841 atgagattat tgagcaaatc agtgaatttt ctaagcgtgt tattttagca gatgccaatt
     3901 tagataaagt tcttagtgca tataacaaac atagagacaa accaatacgt gaacaagcag
     3961 aaaatattat tcatttattt acgttgacga atcttggagc tcccgctgct tttaaatatt
     4021 ttgatacaac aattgatcgt aaacgatata cgtctacaaa agaagtttta gatgccactc
     4081 ttatccatca atccatcact ggtctttatg aaacacgcat tgatttgagt cagctaggag
     4141 gtgacggttc tcccaagaag aagaggaaag tctcgagcgg tggagctgca ggaaattcgg
     4201 gccaccatca ccaccatcac ggcgaacaga aactgatcag cgaagaagat ctgaactagg
     4261 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
     4321 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
     4381 gaactatatc cggatatccc gcaagaggcc cggcagtacc ggcataacca agcctatgcc
     4441 tacagcatcc agggtgacgg tgccgaggat gacgatgagc gcattgttag atttcataca
     4501 cggtgcctga ctgcgttagc aatttaactg tgataaacta ccgcattaaa gctggctcga
     4561 ccaattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     4621 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4681 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4741 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     4801 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     4861 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     4921 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     4981 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg
     5041 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     5101 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     5161 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     5221 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     5281 ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca
     5341 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     5401 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     5461 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     5521 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     5581 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     5641 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5701 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5761 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5821 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5881 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5941 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6001 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     6061 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     6121 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     6181 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     6241 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6301 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6361 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6421 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     6481 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     6541 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     6601 cgcatctgtg cggtatttca caccgcatat atggtgcact ctcagtacaa tctgctctga
     6661 tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg
     6721 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     6781 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     6841 tcaccgaaac gcgcgggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac
     6901 agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct
     6961 ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc acttgatgcc
     7021 tccgtgtaag ggggaatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat
     7081 gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa
     7141 acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg
     7201 cttcgttaat acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat
     7261 ccggaacata atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa
     7321 accgaagacc attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca
     7381 cgttcgctcg cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag
     7441 ccgggtcctc aacgacagga gcacgatcat gcgcacccgt ggccaggacc caacgctgcc
     7501 cgagatgcgc cgcgtgcggc tgctggagat ggcggacgcg atggatatgt tctgccaagg
     7561 gttggtttgc gcattcacag ttctccgcaa gaattgattg gctccaattc ttggagtggt
     7621 gaatccgtta gcgaggtgcc gccggcttcc attcaggtcg aggtggcccg gctccatgca
     7681 ccgcgacgca acgcggggag gcagacaagg tatagggcgg cgcctacaat ccatgccaac
     7741 ccgttccatg tgctcgccga ggcggcataa atcgccgtga cgatcagcgg tccagtgatc
     7801 gaagttaggc tggtaagagc cgcgagcgat ccttgaagct gtccctgatg gtcgtcatct
     7861 acctgcctgg acagcatggc ctgcaacgcg ggcatcccga tgccgccgga agcgagaaga
     7921 atcataatgg ggaaggccat ccagcctcgc gtcgcgaacg ccagcaagac gtagcccagc
     7981 gcgtcggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga
     8041 ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg
     8101 atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc
     8161 acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg
     8221 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat
     8281 cccggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     8341 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     8401 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
     8461 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
     8521 cagcaggcga aaatcctgtt tgatggtggt tgacggcggg atataacatg agctgtcttc
     8581 ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat
     8641 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
     8701 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
     8761 ccgttccgct atcggctgaa tttgattcgc agtgagatat ttatgccagc cagccagacg
     8821 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
     8881 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
     8941 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
     9001 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
     9061 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
     9121 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
     9181 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
     9241 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
     9301 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
     9361 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
     9421 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat
     9481 ggtgtcccgg atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta
     9541 gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc
     9601 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
     9661 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
     9721 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatct
     9781 cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata acaattcccc
//
