LOCUS       sequence-174587-       10073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  D435C/E945C in cysteine-free (C80S/C574S) S. pyogenes Cas9 
            expression plasmid, lobe closure FRET construct.
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: pSHS293
FEATURES             Location/Qualifiers
     source          1..10073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     213..230
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    245..384
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(310..329)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     470..492
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(486..677)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     2135..2154
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2238..2257
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2238..2256
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2345..2367
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2387..2416
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2426..3526
                     /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     3500..3523
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             3581..3601
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             3608..7714
                     /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
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKCNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIESFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDCNDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     primer_bind     complement(7840..7858)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      7854..7901
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(8267..8272,8273..8289,8290..8295))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(8322..8340)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8408..8512
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8513..8581,8582..9373)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRXDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPXAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(8731..8750)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(9544..10073,1..59)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10033..10052
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
       61 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
      121 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
      181 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
      241 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg
      301 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
      361 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
      421 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
      481 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca
      541 tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg
      601 agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt
      661 ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg
      721 ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg
      781 ttactggaac gtcgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa
      841 atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc
      901 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt
      961 tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac
     1021 gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca
     1081 gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc
     1141 cgtggccagg acccaacgct gcccgagatg cgccgcgtgc ggctgctgga gatggcggac
     1201 gcgatggata tgttctgcca agggttggtt tgcgcattca cagttctccg caagaattga
     1261 ttggctccaa ttcttggagt ggtgaatccg ttagcgaggt gccgccggct tccattcagg
     1321 tcgaggtggc ccggctccat gcaccgcgac gcaacgcggg gaggcagaca aggtataggg
     1381 cggcgcctac aatccatgcc aacccgttcc atgtgctcgc cgaggcggca taaatcgccg
     1441 tgacgatcag cggtccaatg atcgaagtta ggctggtaag agccgcgagc gatccttgaa
     1501 gctgtccctg atggtcgtca tctacctgcc tggacagcat ggcctgcaac gcgggcatcc
     1561 cgatgccgcc ggaagcgaga agaatcataa tggggaaggc catccagcct cgcgtcgcga
     1621 acgccagcaa gacgtagccc agcgcgtcgg ccgccatgcc ggcgataatg gcctgcttct
     1681 cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc
     1741 cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga
     1801 aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca
     1861 taagtgcggc gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg
     1921 ctctcaaggg catcggtcga cgctctccct tatgcgactc ctgcattagg aagcagccca
     1981 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg
     2041 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
     2101 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
     2161 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga
     2221 tctcgatccc gcgaaattaa tacgactcac tatagggaga ccacaacggt ttccctctag
     2281 tgccggctcc ggagagctct ttaattaagc ggccgccctg caggactcga gttctagaaa
     2341 taattttgtt taactttaag aaggagatat acatatgaaa tcttctcacc atcaccatca
     2401 ccatcaccat caccatggtt cttctatgaa aatcgaagaa ggtaaactgg taatctggat
     2461 taacggcgat aaaggctata acggtctcgc tgaagtcggt aagaaattcg agaaagatac
     2521 cggaattaaa gtcaccgttg agcatccgga taaactggaa gagaaattcc cacaggttgc
     2581 ggcaactggc gatggccctg acattatctt ctgggcacac gaccgctttg gtggctacgc
     2641 tcaatctggc ctgttggctg aaatcacccc ggacaaagcg ttccaggaca agctgtatcc
     2701 gtttacctgg gatgccgtac gttacaacgg caagctgatt gcttacccga tcgctgttga
     2761 agcgttatcg ctgatttata acaaagatct gctgccgaac ccgccaaaaa cctgggaaga
     2821 gatcccggcg ctggataaag aactgaaagc gaaaggtaag agcgcgctga tgttcaacct
     2881 gcaagaaccg tacttcacct ggccgctgat tgctgctgac gggggttatg cgttcaagta
     2941 tgaaaacggc aagtacgaca ttaaagacgt gggcgtggat aacgctggcg caaaagcggg
     3001 tctgaccttc ctggttgacc tgattaaaaa caaacacatg aatgcagaca ccgattactc
     3061 catcgcagaa gctgccttta ataaaggcga aacagcgatg accatcaacg gcccgtgggc
     3121 atggtccaac atcgacacca gcaaagtgaa ttatggtgta acggtactgc cgaccttcaa
     3181 gggtcaacca tccaaaccgt tcgttggcgt gctgagcgca ggtattaacg ccgccagtcc
     3241 gaacaaagag ctggcaaaag agttcctcga aaactatctg ctgactgatg aaggtctgga
     3301 agcggttaat aaagacaaac cgctgggtgc cgtagcgctg aagtcttacg aggaagagtt
     3361 ggcgaaagat ccacgtattg ccgccactat ggaaaacgcc cagaaaggtg aaatcatgcc
     3421 gaacatcccg cagatgtccg ctttctggta tgccgtgcgt actgcggtga tcaacgccgc
     3481 cagcggtcgt cagactgtcg atgaagccct gaaagacgcg cagactaatt cgagctcgaa
     3541 caacaacaac aataacaata acaacaacct cgggatcgag gaaaacctgt acttccaatc
     3601 caatgcaatg gataagaaat actcaatagg cttagatatc ggcacaaata gcgtcggatg
     3661 ggcggtgatc actgatgaat ataaggttcc gtctaaaaag ttcaaggttc tgggaaatac
     3721 agaccgccac agtatcaaaa aaaatcttat aggggctctt ttatttgaca gtggagagac
     3781 agcggaagcg actcgtctca aacggacagc tcgtagaagg tatacacgtc ggaagaatcg
     3841 tatttcttat ctacaggaga ttttttcaaa tgagatggcg aaagtagatg atagtttctt
     3901 tcatcgactt gaagagtctt ttttggtgga agaagacaag aagcatgaac gtcatcctat
     3961 ttttggaaat atagtagatg aagttgctta tcatgagaaa tatccaacta tctatcatct
     4021 gcgaaaaaaa ttggtagatt ctactgataa agcggatttg cgcttaatct atttggcctt
     4081 agcgcatatg attaagtttc gtggtcattt tttgattgag ggagatttaa atcctgataa
     4141 tagtgatgtg gacaaactat ttatccagtt ggtacaaacc tacaatcaat tatttgaaga
     4201 aaaccctatt aacgcaagtg gagtagatgc taaagcgatt ctttctgcac gattgagtaa
     4261 atcaagacga ttagaaaatc tcattgctca gctccccggt gagaagaaaa atggcttatt
     4321 tgggaatctc attgctttgt cattgggttt gacccctaat tttaaatcaa attttgattt
     4381 ggcagaagat gctaaattac agctttcaaa agatacttac gatgatgatt tagataattt
     4441 attggcgcaa attggagatc aatatgctga tttgtttttg gcagctaaga atttatcaga
     4501 tgctatttta ctttcagata tcctaagagt aaatactgaa ataactaagg ctcccctatc
     4561 agcttcaatg attaaacgct acgatgaaca tcatcaagac ttgactcttt taaaagcttt
     4621 agttcgacaa caacttccag aaaagtataa agaaatcttt tttgatcaat caaaaaacgg
     4681 atatgcaggt tatattgatg ggggagctag ccaagaagaa ttttataaat ttatcaaacc
     4741 aattttagaa aaaatggatg gtactgagga attattggtg aaactaaatc gtgaagattt
     4801 gctgcgcaag caacggacct ttgacaacgg ctctattccc catcaaattc acttgggtga
     4861 gctgcatgct attttgagaa gacaagaaga cttttatcca tttttaaaat gcaatcgtga
     4921 gaagattgaa aaaatcttga cttttcgaat tccttattat gttggtccat tggcgcgtgg
     4981 caatagtcgt tttgcatgga tgactcggaa gtctgaagaa acaattaccc catggaattt
     5041 tgaagaagtt gtcgataaag gtgcttcagc tcaatcattt attgaacgca tgacaaactt
     5101 tgataaaaat cttccaaatg aaaaagtact accaaaacat agtttgcttt atgagtattt
     5161 tacggtttat aacgaattga caaaggtcaa atatgttact gaaggaatgc gaaaaccagc
     5221 atttctttca ggtgaacaga agaaagccat tgttgattta ctcttcaaaa caaatcgaaa
     5281 agtaaccgtt aagcaattaa aagaagatta tttcaaaaaa atagaatctt ttgatagtgt
     5341 tgaaatttca ggagttgaag atagatttaa tgcttcatta ggtacctacc atgatttgct
     5401 aaaaattatt aaagataaag attttttgga taatgaagaa aatgaagata tcttagagga
     5461 tattgtttta acattgacct tatttgaaga tagggagatg attgaggaaa gacttaaaac
     5521 atatgctcac ctctttgatg ataaggtgat gaaacagctt aaacgtcgcc gttatactgg
     5581 ttggggacgt ttgtctcgaa aattgattaa tggtattagg gataagcaat ctggcaaaac
     5641 aatattagat tttttgaaat cagatggttt tgccaatcgc aattttatgc agctgatcca
     5701 tgatgatagt ttgacattta aagaagacat tcaaaaagca caagtgtctg gacaaggcga
     5761 tagtttacat gaacatattg caaatttagc tggtagccct gctattaaaa aaggtatttt
     5821 acagactgta aaagttgttg atgaattggt caaagtaatg gggcggcata agccagaaaa
     5881 tatcgttatt gaaatggcac gtgaaaatca gacaactcaa aagggccaga aaaattcgcg
     5941 agagcgtatg aaacgaatcg aagaaggtat caaagaatta ggaagtcaga ttcttaaaga
     6001 gcatcctgtt gaaaatactc aattgcaaaa tgaaaagctc tatctctatt atctccaaaa
     6061 tggaagagac atgtatgtgg accaagaatt agatattaat cgtttaagtg attatgatgt
     6121 cgatcacatt gttccacaaa gtttccttaa agacgattca atagacaata aggtcttaac
     6181 gcgttctgat aaaaatcgtg gtaaatcgga taacgttcca agtgaagaag tagtcaaaaa
     6241 gatgaaaaac tattggagac aacttctaaa cgccaagtta atcactcaac gtaagtttga
     6301 taatttaacg aaagctgaac gtggaggttt gagtgaactt gataaagctg gttttatcaa
     6361 acgccaattg gttgaaactc gccaaatcac taagcatgtg gcacaaattt tggatagtcg
     6421 catgaatact aaatacgatt gcaatgataa acttattcga gaggttaaag tgattacctt
     6481 aaaatctaaa ttagtttctg acttccgaaa agatttccaa ttctataaag tacgtgagat
     6541 taacaattac catcatgccc atgatgcgta tctaaatgcc gtcgttggaa ctgctttgat
     6601 taagaaatat ccaaaacttg aatcggagtt tgtctatggt gattataaag tttatgatgt
     6661 tcgtaaaatg attgctaagt ctgagcaaga aataggcaaa gcaaccgcaa aatatttctt
     6721 ttactctaat atcatgaact tcttcaaaac agaaattaca cttgcaaatg gagagattcg
     6781 caaacgccct ctaatcgaaa ctaatgggga aactggagaa attgtctggg ataaagggcg
     6841 agattttgcc acagtgcgca aagtattgtc catgccccaa gtcaatattg tcaagaaaac
     6901 agaagtacag acaggcggat tctccaagga gtcaatttta ccaaaaagaa attcggacaa
     6961 gcttattgct cgtaaaaaag actgggatcc aaaaaaatat ggtggttttg atagtccaac
     7021 ggtagcttat tcagtcctag tggttgctaa ggtggaaaaa gggaaatcga agaagttaaa
     7081 atccgttaaa gagttactag ggatcacaat tatggaaaga agttcctttg aaaaaaatcc
     7141 gattgacttt ttagaagcta aaggatataa ggaagttaaa aaagacttaa tcattaaact
     7201 acctaaatat agtctttttg agttagaaaa cggtcgtaaa cggatgctgg ctagtgccgg
     7261 agaattacaa aaaggaaatg agctggctct gccaagcaaa tatgtgaatt ttttatattt
     7321 agctagtcat tatgaaaagt tgaagggtag tccagaagat aacgaacaaa aacaattgtt
     7381 tgtggagcag cataagcatt atttagatga gattattgag caaatcagtg aattttctaa
     7441 gcgtgttatt ttagcagatg ccaatttaga taaagttctt agtgcatata acaaacatag
     7501 agacaaacca atacgtgaac aagcagaaaa tattattcat ttatttacgt tgacgaatct
     7561 tggagctccc gctgctttta aatattttga tacaacaatt gatcgtaaac gatatacgtc
     7621 tacaaaagaa gttttagatg ccactcttat ccatcaatcc atcactggtc tttatgaaac
     7681 acgcattgat ttgagtcagc taggaggtga ctaataacat tggaagtgga taacggatcc
     7741 gcgatcgcgg cgcgccacct ggtggccggc cggtaccacg cgtgcgcgct gatccggctg
     7801 ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat
     7861 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat
     7921 ccggatatcc acaggacggg tgtggtcgcc atgatcgcgt agtcgatagt ggctccaagt
     7981 agcgaagcga gcaggactgg gcggcggcca aagcggtcgg acagtgctcc gagaacgggt
     8041 gcgcatagaa attgcatcaa cgcatatagc gctagcagca cgccatagtg actggcgatg
     8101 ctgtcggaat ggacgatatc ccgcaagagg cccggcagta ccggcataac caagcctatg
     8161 cctacagcat ccagggtgac ggtgccgagg atgacgatga gcgcattgtt agatttcata
     8221 cacggtgcct gactgcgtta gcaatttaac tgtgataaac taccgcatta aagcttatcg
     8281 atgataagct gtcaaacatg agaattcttg aagacgaaag ggcctcgtga tacgcctatt
     8341 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     8401 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     8461 catgagacaa taaccctgat aaatgcttca ataacattga aaaaggaaga gtatgagtat
     8521 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     8581 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     8641 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     8701 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtrttga
     8761 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     8821 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     8881 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     8941 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     9001 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgyagc
     9061 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     9121 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     9181 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     9241 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     9301 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     9361 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     9421 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     9481 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     9541 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     9601 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     9661 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     9721 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     9781 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     9841 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     9901 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     9961 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
    10021 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gcc
//
