LOCUS       sequence-189236-        9232 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Cas9-6xHis in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9232)
  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 9232)
  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-Cas9-6xHis
FEATURES             Location/Qualifiers
     source          1..9232
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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             42..64
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             71..4174
                     /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             71..4174
                     /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             4175..4192
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4263..4281)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4277..4324
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4361..4816
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4448..4467)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4658..4679
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(4931..5722,5723..5791))
                     /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     5554..5573
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5792..5884)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5965..6553
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6454..6473
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6707..6724
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6739..6878
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6804..6823)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6964..6986
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6980..7171)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(7743..8825)
                     /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     8787..8806
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(8826..8903)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     9109..9128
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     join(9216..9232,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(9216..9232,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
       61 gagatatacc atggataaga aatactcaat aggcttagat atcggcacaa atagcgtcgg
      121 atgggcggtg atcactgatg aatataaggt tccgtctaaa aagttcaagg ttctgggaaa
      181 tacagaccgc cacagtatca aaaaaaatct tataggggct cttttatttg acagtggaga
      241 gacagcggaa gcgactcgtc tcaaacggac agctcgtaga aggtatacac gtcggaagaa
      301 tcgtatttgt tatctacagg agattttttc aaatgagatg gcgaaagtag atgatagttt
      361 ctttcatcga cttgaagagt cttttttggt ggaagaagac aagaagcatg aacgtcatcc
      421 tatttttgga aatatagtag atgaagttgc ttatcatgag aaatatccaa ctatctatca
      481 tctgcgaaaa aaattggtag attctactga taaagcggat ttgcgcttaa tctatttggc
      541 cttagcgcat atgattaagt ttcgtggtca ttttttgatt gagggagatt taaatcctga
      601 taatagtgat gtggacaaac tatttatcca gttggtacaa acctacaatc aattatttga
      661 agaaaaccct attaacgcaa gtggagtaga tgctaaagcg attctttctg cacgattgag
      721 taaatcaaga cgattagaaa atctcattgc tcagctcccc ggtgagaaga aaaatggctt
      781 atttgggaat ctcattgctt tgtcattggg tttgacccct aattttaaat caaattttga
      841 tttggcagaa gatgctaaat tacagctttc aaaagatact tacgatgatg atttagataa
      901 tttattggcg caaattggag atcaatatgc tgatttgttt ttggcagcta agaatttatc
      961 agatgctatt ttactttcag atatcctaag agtaaatact gaaataacta aggctcccct
     1021 atcagcttca atgattaaac gctacgatga acatcatcaa gacttgactc ttttaaaagc
     1081 tttagttcga caacaacttc cagaaaagta taaagaaatc ttttttgatc aatcaaaaaa
     1141 cggatatgca ggttatattg atgggggagc tagccaagaa gaattttata aatttatcaa
     1201 accaatttta gaaaaaatgg atggtactga ggaattattg gtgaaactaa atcgtgaaga
     1261 tttgctgcgc aagcaacgga cctttgacaa cggctctatt ccccatcaaa ttcacttggg
     1321 tgagctgcat gctattttga gaagacaaga agacttttat ccatttttaa aagacaatcg
     1381 tgagaagatt gaaaaaatct tgacttttcg aattccttat tatgttggtc cattggcgcg
     1441 tggcaatagt cgttttgcat ggatgactcg gaagtctgaa gaaacaatta ccccatggaa
     1501 ttttgaagaa gttgtcgata aaggtgcttc agctcaatca tttattgaac gcatgacaaa
     1561 ctttgataaa aatcttccaa atgaaaaagt actaccaaaa catagtttgc tttatgagta
     1621 ttttacggtt tataacgaat tgacaaaggt caaatatgtt actgaaggaa tgcgaaaacc
     1681 agcatttctt tcaggtgaac agaagaaagc cattgttgat ttactcttca aaacaaatcg
     1741 aaaagtaacc gttaagcaat taaaagaaga ttatttcaaa aaaatagaat gttttgatag
     1801 tgttgaaatt tcaggagttg aagatagatt taatgcttca ttaggtacct accatgattt
     1861 gctaaaaatt attaaagata aagatttttt ggataatgaa gaaaatgaag atatcttaga
     1921 ggatattgtt ttaacattga ccttatttga agatagggag atgattgagg aaagacttaa
     1981 aacatatgct cacctctttg atgataaggt gatgaaacag cttaaacgtc gccgttatac
     2041 tggttgggga cgtttgtctc gaaaattgat taatggtatt agggataagc aatctggcaa
     2101 aacaatatta gattttttga aatcagatgg ttttgccaat cgcaatttta tgcagctgat
     2161 ccatgatgat agtttgacat ttaaagaaga cattcaaaaa gcacaagtgt ctggacaagg
     2221 cgatagttta catgaacata ttgcaaattt agctggtagc cctgctatta aaaaaggtat
     2281 tttacagact gtaaaagttg ttgatgaatt ggtcaaagta atggggcggc ataagccaga
     2341 aaatatcgtt attgaaatgg cacgtgaaaa tcagacaact caaaagggcc agaaaaattc
     2401 gcgagagcgt atgaaacgaa tcgaagaagg tatcaaagaa ttaggaagtc agattcttaa
     2461 agagcatcct gttgaaaata ctcaattgca aaatgaaaag ctctatctct attatctcca
     2521 aaatggaaga gacatgtatg tggaccaaga attagatatt aatcgtttaa gtgattatga
     2581 tgtcgatcac attgttccac aaagtttcct taaagacgat tcaatagaca ataaggtctt
     2641 aacgcgttct gataaaaatc gtggtaaatc ggataacgtt ccaagtgaag aagtagtcaa
     2701 aaagatgaaa aactattgga gacaacttct aaacgccaag ttaatcactc aacgtaagtt
     2761 tgataattta acgaaagctg aacgtggagg tttgagtgaa cttgataaag ctggttttat
     2821 caaacgccaa ttggttgaaa ctcgccaaat cactaagcat gtggcacaaa ttttggatag
     2881 tcgcatgaat actaaatacg atgaaaatga taaacttatt cgagaggtta aagtgattac
     2941 cttaaaatct aaattagttt ctgacttccg aaaagatttc caattctata aagtacgtga
     3001 gattaacaat taccatcatg cccatgatgc gtatctaaat gccgtcgttg gaactgcttt
     3061 gattaagaaa tatccaaaac ttgaatcgga gtttgtctat ggtgattata aagtttatga
     3121 tgttcgtaaa atgattgcta agtctgagca agaaataggc aaagcaaccg caaaatattt
     3181 cttttactct aatatcatga acttcttcaa aacagaaatt acacttgcaa atggagagat
     3241 tcgcaaacgc cctctaatcg aaactaatgg ggaaactgga gaaattgtct gggataaagg
     3301 gcgagatttt gccacagtgc gcaaagtatt gtccatgccc caagtcaata ttgtcaagaa
     3361 aacagaagta cagacaggcg gattctccaa ggagtcaatt ttaccaaaaa gaaattcgga
     3421 caagcttatt gctcgtaaaa aagactggga tccaaaaaaa tatggtggtt ttgatagtcc
     3481 aacggtagct tattcagtcc tagtggttgc taaggtggaa aaagggaaat cgaagaagtt
     3541 aaaatccgtt aaagagttac tagggatcac aattatggaa agaagttcct ttgaaaaaaa
     3601 tccgattgac tttttagaag ctaaaggata taaggaagtt aaaaaagact taatcattaa
     3661 actacctaaa tatagtcttt ttgagttaga aaacggtcgt aaacggatgc tggctagtgc
     3721 cggagaatta caaaaaggaa atgagctggc tctgccaagc aaatatgtga attttttata
     3781 tttagctagt cattatgaaa agttgaaggg tagtccagaa gataacgaac aaaaacaatt
     3841 gtttgtggag cagcataagc attatttaga tgagattatt gagcaaatca gtgaattttc
     3901 taagcgtgtt attttagcag atgccaattt agataaagtt cttagtgcat ataacaaaca
     3961 tagagacaaa ccaatacgtg aacaagcaga aaatattatt catttattta cgttgacgaa
     4021 tcttggagct cccgctgctt ttaaatattt tgatacaaca attgatcgta aacgatatac
     4081 gtctacaaaa gaagttttag atgccactct tatccatcaa tccatcactg gtctttatga
     4141 aacacgcatt gatttgagtc agctaggagg tgaccatcac caccaccatc actaatgata
     4201 atttgaacgc cagcacatgg actcgtctac tagcgcagct taattaacct aggctgctgc
     4261 caccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
     4321 tttgctgaaa ggaggaacta tatccggatt ggcgaatggg acgcgccctg tagcggcgca
     4381 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     4441 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     4501 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     4561 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     4621 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     4681 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     4741 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     4801 ttaacgttta caatttctgg cggcacgatg gcatgagatt atcaaaaagg atcttcacct
     4861 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     4921 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     4981 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5041 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5101 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5161 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5221 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     5281 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     5341 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     5401 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     5461 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     5521 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     5581 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     5641 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     5701 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     5761 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatca tgattgaagc
     5821 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5881 caaataggtc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5941 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     6001 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     6061 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     6121 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     6181 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6241 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6301 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6361 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6421 cggaacagga gagcgcacga gggagctccc agggggaaac gcctggtatc tttatagtcc
     6481 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6541 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6601 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     6661 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     6721 cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     6781 acaccgcaat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     6841 acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg
     6901 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     6961 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc
     7021 tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg
     7081 cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca
     7141 tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt
     7201 tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg
     7261 atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc
     7321 gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg
     7381 ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg
     7441 ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg
     7501 ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt
     7561 cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca
     7621 cgatcatgct agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca
     7681 agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc
     7741 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     7801 aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt
     7861 gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg
     7921 tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata
     7981 taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatgtccgc accaacgcgc
     8041 agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc
     8101 atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg
     8161 gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta
     8221 tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg
     8281 atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg
     8341 gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca
     8401 ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc
     8461 agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg
     8521 cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc
     8581 gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc
     8641 aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc
     8701 atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg
     8761 cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt
     8821 ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag
     8881 gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat
     8941 taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc
     9001 atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc
     9061 gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc
     9121 gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc
     9181 gtagaggatc gagatcgatc tcgatcccgc gaaattaata cgactcacta ta
//
