LOCUS       sequence-174283-       10073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cysteine-free (C80S/C574S) S. pyogenes Cas9 expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10073)
  AUTHORS   Sternberg SH, LaFrance B, Kaplan M, Doudna JA
  TITLE     Conformational control of DNA target cleavage by CRISPR-Cas9.
  JOURNAL   Nature. 2015 Nov 5;527(7576):110-3. doi: 10.1038/nature15544. Epub 
            2015 Oct 28.
  PUBMED    26524520
REFERENCE   2  (bases 1 to 10073)
  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/nature15544"; journalName: "Nature"; date: "2015-11-5- 5"; 
            volume: "527"; issue: "7576"; pages: "110-3"
COMMENT     Sequence Label: pSHS212
FEATURES             Location/Qualifiers
     source          1..10073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      135..723
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     624..643
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     877..894
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    909..1048
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(974..993)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1134..1156
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1150..1341)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     2799..2818
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2902..2921
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2902..2920
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             3009..3031
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3051..3080
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             3090..4190
                     /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="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     4164..4187
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             4245..4265
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             4272..8378
                     /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
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRISYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIESFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     primer_bind     complement(8504..8522)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      8518..8565
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(8931..8936,8937..8953,8954..8959))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(8986..9004)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9072..9176
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9177..9245,9246..10037)
                     /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(9395..9414)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
       61 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
      121 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
      181 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
      241 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
      301 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
      361 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
      421 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
      481 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
      541 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
      601 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
      661 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
      721 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
      781 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
      841 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
      901 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaat
      961 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct
     1021 atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc
     1081 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     1141 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag
     1201 ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc
     1261 gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc
     1321 ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt
     1381 aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc
     1441 ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag
     1501 aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg
     1561 tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg
     1621 cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc
     1681 agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta
     1741 accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg
     1801 cacccgtggc caggacccaa cgctgcccga gatgcgccgc gtgcggctgc tggagatggc
     1861 ggacgcgatg gatatgttct gccaagggtt ggtttgcgca ttcacagttc tccgcaagaa
     1921 ttgattggct ccaattcttg gagtggtgaa tccgttagcg aggtgccgcc ggcttccatt
     1981 caggtcgagg tggcccggct ccatgcaccg cgacgcaacg cggggaggca gacaaggtat
     2041 agggcggcgc ctacaatcca tgccaacccg ttccatgtgc tcgccgaggc ggcataaatc
     2101 gccgtgacga tcagcggtcc aatgatcgaa gttaggctgg taagagccgc gagcgatcct
     2161 tgaagctgtc cctgatggtc gtcatctacc tgcctggaca gcatggcctg caacgcgggc
     2221 atcccgatgc cgccggaagc gagaagaatc ataatgggga aggccatcca gcctcgcgtc
     2281 gcgaacgcca gcaagacgta gcccagcgcg tcggccgcca tgccggcgat aatggcctgc
     2341 ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag
     2401 attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg
     2461 ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca
     2521 gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg
     2581 aaggctctca agggcatcgg tcgacgctct cccttatgcg actcctgcat taggaagcag
     2641 cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag
     2701 atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg
     2761 ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg
     2821 ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc
     2881 gagatctcga tcccgcgaaa ttaatacgac tcactatagg gagaccacaa cggtttccct
     2941 ctagtgccgg ctccggagag ctctttaatt aagcggccgc cctgcaggac tcgagttcta
     3001 gaaataattt tgtttaactt taagaaggag atatacatat gaaatcttct caccatcacc
     3061 atcaccatca ccatcaccat ggttcttcta tgaaaatcga agaaggtaaa ctggtaatct
     3121 ggattaacgg cgataaaggc tataacggtc tcgctgaagt cggtaagaaa ttcgagaaag
     3181 ataccggaat taaagtcacc gttgagcatc cggataaact ggaagagaaa ttcccacagg
     3241 ttgcggcaac tggcgatggc cctgacatta tcttctgggc acacgaccgc tttggtggct
     3301 acgctcaatc tggcctgttg gctgaaatca ccccggacaa agcgttccag gacaagctgt
     3361 atccgtttac ctgggatgcc gtacgttaca acggcaagct gattgcttac ccgatcgctg
     3421 ttgaagcgtt atcgctgatt tataacaaag atctgctgcc gaacccgcca aaaacctggg
     3481 aagagatccc ggcgctggat aaagaactga aagcgaaagg taagagcgcg ctgatgttca
     3541 acctgcaaga accgtacttc acctggccgc tgattgctgc tgacgggggt tatgcgttca
     3601 agtatgaaaa cggcaagtac gacattaaag acgtgggcgt ggataacgct ggcgcaaaag
     3661 cgggtctgac cttcctggtt gacctgatta aaaacaaaca catgaatgca gacaccgatt
     3721 actccatcgc agaagctgcc tttaataaag gcgaaacagc gatgaccatc aacggcccgt
     3781 gggcatggtc caacatcgac accagcaaag tgaattatgg tgtaacggta ctgccgacct
     3841 tcaagggtca accatccaaa ccgttcgttg gcgtgctgag cgcaggtatt aacgccgcca
     3901 gtccgaacaa agagctggca aaagagttcc tcgaaaacta tctgctgact gatgaaggtc
     3961 tggaagcggt taataaagac aaaccgctgg gtgccgtagc gctgaagtct tacgaggaag
     4021 agttggcgaa agatccacgt attgccgcca ctatggaaaa cgcccagaaa ggtgaaatca
     4081 tgccgaacat cccgcagatg tccgctttct ggtatgccgt gcgtactgcg gtgatcaacg
     4141 ccgccagcgg tcgtcagact gtcgatgaag ccctgaaaga cgcgcagact aattcgagct
     4201 cgaacaacaa caacaataac aataacaaca acctcgggat cgaggaaaac ctgtacttcc
     4261 aatccaatgc aatggataag aaatactcaa taggcttaga tatcggcaca aatagcgtcg
     4321 gatgggcggt gatcactgat gaatataagg ttccgtctaa aaagttcaag gttctgggaa
     4381 atacagaccg ccacagtatc aaaaaaaatc ttataggggc tcttttattt gacagtggag
     4441 agacagcgga agcgactcgt ctcaaacgga cagctcgtag aaggtataca cgtcggaaga
     4501 atcgtatttc ttatctacag gagatttttt caaatgagat ggcgaaagta gatgatagtt
     4561 tctttcatcg acttgaagag tcttttttgg tggaagaaga caagaagcat gaacgtcatc
     4621 ctatttttgg aaatatagta gatgaagttg cttatcatga gaaatatcca actatctatc
     4681 atctgcgaaa aaaattggta gattctactg ataaagcgga tttgcgctta atctatttgg
     4741 ccttagcgca tatgattaag tttcgtggtc attttttgat tgagggagat ttaaatcctg
     4801 ataatagtga tgtggacaaa ctatttatcc agttggtaca aacctacaat caattatttg
     4861 aagaaaaccc tattaacgca agtggagtag atgctaaagc gattctttct gcacgattga
     4921 gtaaatcaag acgattagaa aatctcattg ctcagctccc cggtgagaag aaaaatggct
     4981 tatttgggaa tctcattgct ttgtcattgg gtttgacccc taattttaaa tcaaattttg
     5041 atttggcaga agatgctaaa ttacagcttt caaaagatac ttacgatgat gatttagata
     5101 atttattggc gcaaattgga gatcaatatg ctgatttgtt tttggcagct aagaatttat
     5161 cagatgctat tttactttca gatatcctaa gagtaaatac tgaaataact aaggctcccc
     5221 tatcagcttc aatgattaaa cgctacgatg aacatcatca agacttgact cttttaaaag
     5281 ctttagttcg acaacaactt ccagaaaagt ataaagaaat cttttttgat caatcaaaaa
     5341 acggatatgc aggttatatt gatgggggag ctagccaaga agaattttat aaatttatca
     5401 aaccaatttt agaaaaaatg gatggtactg aggaattatt ggtgaaacta aatcgtgaag
     5461 atttgctgcg caagcaacgg acctttgaca acggctctat tccccatcaa attcacttgg
     5521 gtgagctgca tgctattttg agaagacaag aagactttta tccattttta aaagacaatc
     5581 gtgagaagat tgaaaaaatc ttgacttttc gaattcctta ttatgttggt ccattggcgc
     5641 gtggcaatag tcgttttgca tggatgactc ggaagtctga agaaacaatt accccatgga
     5701 attttgaaga agttgtcgat aaaggtgctt cagctcaatc atttattgaa cgcatgacaa
     5761 actttgataa aaatcttcca aatgaaaaag tactaccaaa acatagtttg ctttatgagt
     5821 attttacggt ttataacgaa ttgacaaagg tcaaatatgt tactgaagga atgcgaaaac
     5881 cagcatttct ttcaggtgaa cagaagaaag ccattgttga tttactcttc aaaacaaatc
     5941 gaaaagtaac cgttaagcaa ttaaaagaag attatttcaa aaaaatagaa tcttttgata
     6001 gtgttgaaat ttcaggagtt gaagatagat ttaatgcttc attaggtacc taccatgatt
     6061 tgctaaaaat tattaaagat aaagattttt tggataatga agaaaatgaa gatatcttag
     6121 aggatattgt tttaacattg accttatttg aagataggga gatgattgag gaaagactta
     6181 aaacatatgc tcacctcttt gatgataagg tgatgaaaca gcttaaacgt cgccgttata
     6241 ctggttgggg acgtttgtct cgaaaattga ttaatggtat tagggataag caatctggca
     6301 aaacaatatt agattttttg aaatcagatg gttttgccaa tcgcaatttt atgcagctga
     6361 tccatgatga tagtttgaca tttaaagaag acattcaaaa agcacaagtg tctggacaag
     6421 gcgatagttt acatgaacat attgcaaatt tagctggtag ccctgctatt aaaaaaggta
     6481 ttttacagac tgtaaaagtt gttgatgaat tggtcaaagt aatggggcgg cataagccag
     6541 aaaatatcgt tattgaaatg gcacgtgaaa atcagacaac tcaaaagggc cagaaaaatt
     6601 cgcgagagcg tatgaaacga atcgaagaag gtatcaaaga attaggaagt cagattctta
     6661 aagagcatcc tgttgaaaat actcaattgc aaaatgaaaa gctctatctc tattatctcc
     6721 aaaatggaag agacatgtat gtggaccaag aattagatat taatcgttta agtgattatg
     6781 atgtcgatca cattgttcca caaagtttcc ttaaagacga ttcaatagac aataaggtct
     6841 taacgcgttc tgataaaaat cgtggtaaat cggataacgt tccaagtgaa gaagtagtca
     6901 aaaagatgaa aaactattgg agacaacttc taaacgccaa gttaatcact caacgtaagt
     6961 ttgataattt aacgaaagct gaacgtggag gtttgagtga acttgataaa gctggtttta
     7021 tcaaacgcca attggttgaa actcgccaaa tcactaagca tgtggcacaa attttggata
     7081 gtcgcatgaa tactaaatac gatgaaaatg ataaacttat tcgagaggtt aaagtgatta
     7141 ccttaaaatc taaattagtt tctgacttcc gaaaagattt ccaattctat aaagtacgtg
     7201 agattaacaa ttaccatcat gcccatgatg cgtatctaaa tgccgtcgtt ggaactgctt
     7261 tgattaagaa atatccaaaa cttgaatcgg agtttgtcta tggtgattat aaagtttatg
     7321 atgttcgtaa aatgattgct aagtctgagc aagaaatagg caaagcaacc gcaaaatatt
     7381 tcttttactc taatatcatg aacttcttca aaacagaaat tacacttgca aatggagaga
     7441 ttcgcaaacg ccctctaatc gaaactaatg gggaaactgg agaaattgtc tgggataaag
     7501 ggcgagattt tgccacagtg cgcaaagtat tgtccatgcc ccaagtcaat attgtcaaga
     7561 aaacagaagt acagacaggc ggattctcca aggagtcaat tttaccaaaa agaaattcgg
     7621 acaagcttat tgctcgtaaa aaagactggg atccaaaaaa atatggtggt tttgatagtc
     7681 caacggtagc ttattcagtc ctagtggttg ctaaggtgga aaaagggaaa tcgaagaagt
     7741 taaaatccgt taaagagtta ctagggatca caattatgga aagaagttcc tttgaaaaaa
     7801 atccgattga ctttttagaa gctaaaggat ataaggaagt taaaaaagac ttaatcatta
     7861 aactacctaa atatagtctt tttgagttag aaaacggtcg taaacggatg ctggctagtg
     7921 ccggagaatt acaaaaagga aatgagctgg ctctgccaag caaatatgtg aattttttat
     7981 atttagctag tcattatgaa aagttgaagg gtagtccaga agataacgaa caaaaacaat
     8041 tgtttgtgga gcagcataag cattatttag atgagattat tgagcaaatc agtgaatttt
     8101 ctaagcgtgt tattttagca gatgccaatt tagataaagt tcttagtgca tataacaaac
     8161 atagagacaa accaatacgt gaacaagcag aaaatattat tcatttattt acgttgacga
     8221 atcttggagc tcccgctgct tttaaatatt ttgatacaac aattgatcgt aaacgatata
     8281 cgtctacaaa agaagtttta gatgccactc ttatccatca atccatcact ggtctttatg
     8341 aaacacgcat tgatttgagt cagctaggag gtgactaata acattggaag tggataacgg
     8401 atccgcgatc gcggcgcgcc acctggtggc cggccggtac cacgcgtgcg cgctgatccg
     8461 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta
     8521 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact
     8581 atatccggat atccacagga cgggtgtggt cgccatgatc gcgtagtcga tagtggctcc
     8641 aagtagcgaa gcgagcagga ctgggcggcg gccaaagcgg tcggacagtg ctccgagaac
     8701 gggtgcgcat agaaattgca tcaacgcata tagcgctagc agcacgccat agtgactggc
     8761 gatgctgtcg gaatggacga tatcccgcaa gaggcccggc agtaccggca taaccaagcc
     8821 tatgcctaca gcatccaggg tgacggtgcc gaggatgacg atgagcgcat tgttagattt
     8881 catacacggt gcctgactgc gttagcaatt taactgtgat aaactaccgc attaaagctt
     8941 atcgatgata agctgtcaaa catgagaatt cttgaagacg aaagggcctc gtgatacgcc
     9001 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     9061 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     9121 cgctcatgag acaataaccc tgataaatgc ttcaataaca ttgaaaaagg aagagtatga
     9181 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     9241 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     9301 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     9361 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
     9421 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     9481 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     9541 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     9601 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     9661 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     9721 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     9781 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     9841 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     9901 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     9961 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
    10021 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag att
//
