LOCUS       sequence-174285-       10073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  S867C/S355C in cysteine-free (C80S/C574S) S. pyogenes Cas9 
            expression plasmid, HNH-1 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: pSHS306 - Bacterial expression plasmid for SpC...
FEATURES             Location/Qualifiers
     source          1..10073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     63..82
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     166..185
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        166..184
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             273..295
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             315..344
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             354..1454
                     /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     1428..1451
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1509..1529
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1536..5642
                     /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
                     QQLPEKYKEIFFDQCKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIESFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKCDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     primer_bind     complement(5768..5786)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5782..5829
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(6195..6200,6201..6217,6218..6223))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(6250..6268)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6336..6440
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6441..6509,6510..7301)
                     /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(6659..6678)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7472..8060
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7961..7980
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8214..8231
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    8246..8385
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(8311..8330)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8471..8493
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(8487..8678)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca
       61 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc
      121 acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc gaaattaata cgactcacta
      181 tagggagacc acaacggttt ccctctagtg ccggctccgg agagctcttt aattaagcgg
      241 ccgccctgca ggactcgagt tctagaaata attttgttta actttaagaa ggagatatac
      301 atatgaaatc ttctcaccat caccatcacc atcaccatca ccatggttct tctatgaaaa
      361 tcgaagaagg taaactggta atctggatta acggcgataa aggctataac ggtctcgctg
      421 aagtcggtaa gaaattcgag aaagataccg gaattaaagt caccgttgag catccggata
      481 aactggaaga gaaattccca caggttgcgg caactggcga tggccctgac attatcttct
      541 gggcacacga ccgctttggt ggctacgctc aatctggcct gttggctgaa atcaccccgg
      601 acaaagcgtt ccaggacaag ctgtatccgt ttacctggga tgccgtacgt tacaacggca
      661 agctgattgc ttacccgatc gctgttgaag cgttatcgct gatttataac aaagatctgc
      721 tgccgaaccc gccaaaaacc tgggaagaga tcccggcgct ggataaagaa ctgaaagcga
      781 aaggtaagag cgcgctgatg ttcaacctgc aagaaccgta cttcacctgg ccgctgattg
      841 ctgctgacgg gggttatgcg ttcaagtatg aaaacggcaa gtacgacatt aaagacgtgg
      901 gcgtggataa cgctggcgca aaagcgggtc tgaccttcct ggttgacctg attaaaaaca
      961 aacacatgaa tgcagacacc gattactcca tcgcagaagc tgcctttaat aaaggcgaaa
     1021 cagcgatgac catcaacggc ccgtgggcat ggtccaacat cgacaccagc aaagtgaatt
     1081 atggtgtaac ggtactgccg accttcaagg gtcaaccatc caaaccgttc gttggcgtgc
     1141 tgagcgcagg tattaacgcc gccagtccga acaaagagct ggcaaaagag ttcctcgaaa
     1201 actatctgct gactgatgaa ggtctggaag cggttaataa agacaaaccg ctgggtgccg
     1261 tagcgctgaa gtcttacgag gaagagttgg cgaaagatcc acgtattgcc gccactatgg
     1321 aaaacgccca gaaaggtgaa atcatgccga acatcccgca gatgtccgct ttctggtatg
     1381 ccgtgcgtac tgcggtgatc aacgccgcca gcggtcgtca gactgtcgat gaagccctga
     1441 aagacgcgca gactaattcg agctcgaaca acaacaacaa taacaataac aacaacctcg
     1501 ggatcgagga aaacctgtac ttccaatcca atgcaatgga taagaaatac tcaataggct
     1561 tagatatcgg cacaaatagc gtcggatggg cggtgatcac tgatgaatat aaggttccgt
     1621 ctaaaaagtt caaggttctg ggaaatacag accgccacag tatcaaaaaa aatcttatag
     1681 gggctctttt atttgacagt ggagagacag cggaagcgac tcgtctcaaa cggacagctc
     1741 gtagaaggta tacacgtcgg aagaatcgta tttcttatct acaggagatt ttttcaaatg
     1801 agatggcgaa agtagatgat agtttctttc atcgacttga agagtctttt ttggtggaag
     1861 aagacaagaa gcatgaacgt catcctattt ttggaaatat agtagatgaa gttgcttatc
     1921 atgagaaata tccaactatc tatcatctgc gaaaaaaatt ggtagattct actgataaag
     1981 cggatttgcg cttaatctat ttggccttag cgcatatgat taagtttcgt ggtcattttt
     2041 tgattgaggg agatttaaat cctgataata gtgatgtgga caaactattt atccagttgg
     2101 tacaaaccta caatcaatta tttgaagaaa accctattaa cgcaagtgga gtagatgcta
     2161 aagcgattct ttctgcacga ttgagtaaat caagacgatt agaaaatctc attgctcagc
     2221 tccccggtga gaagaaaaat ggcttatttg ggaatctcat tgctttgtca ttgggtttga
     2281 cccctaattt taaatcaaat tttgatttgg cagaagatgc taaattacag ctttcaaaag
     2341 atacttacga tgatgattta gataatttat tggcgcaaat tggagatcaa tatgctgatt
     2401 tgtttttggc agctaagaat ttatcagatg ctattttact ttcagatatc ctaagagtaa
     2461 atactgaaat aactaaggct cccctatcag cttcaatgat taaacgctac gatgaacatc
     2521 atcaagactt gactctttta aaagctttag ttcgacaaca acttccagaa aagtataaag
     2581 aaatcttttt tgatcaatgt aaaaacggat atgcaggtta tattgatggg ggagctagcc
     2641 aagaagaatt ttataaattt atcaaaccaa ttttagaaaa aatggatggt actgaggaat
     2701 tattggtgaa actaaatcgt gaagatttgc tgcgcaagca acggaccttt gacaacggct
     2761 ctattcccca tcaaattcac ttgggtgagc tgcatgctat tttgagaaga caagaagact
     2821 tttatccatt tttaaaagac aatcgtgaga agattgaaaa aatcttgact tttcgaattc
     2881 cttattatgt tggtccattg gcgcgtggca atagtcgttt tgcatggatg actcggaagt
     2941 ctgaagaaac aattacccca tggaattttg aagaagttgt cgataaaggt gcttcagctc
     3001 aatcatttat tgaacgcatg acaaactttg ataaaaatct tccaaatgaa aaagtactac
     3061 caaaacatag tttgctttat gagtatttta cggtttataa cgaattgaca aaggtcaaat
     3121 atgttactga aggaatgcga aaaccagcat ttctttcagg tgaacagaag aaagccattg
     3181 ttgatttact cttcaaaaca aatcgaaaag taaccgttaa gcaattaaaa gaagattatt
     3241 tcaaaaaaat agaatctttt gatagtgttg aaatttcagg agttgaagat agatttaatg
     3301 cttcattagg tacctaccat gatttgctaa aaattattaa agataaagat tttttggata
     3361 atgaagaaaa tgaagatatc ttagaggata ttgttttaac attgacctta tttgaagata
     3421 gggagatgat tgaggaaaga cttaaaacat atgctcacct ctttgatgat aaggtgatga
     3481 aacagcttaa acgtcgccgt tatactggtt ggggacgttt gtctcgaaaa ttgattaatg
     3541 gtattaggga taagcaatct ggcaaaacaa tattagattt tttgaaatca gatggttttg
     3601 ccaatcgcaa ttttatgcag ctgatccatg atgatagttt gacatttaaa gaagacattc
     3661 aaaaagcaca agtgtctgga caaggcgata gtttacatga acatattgca aatttagctg
     3721 gtagccctgc tattaaaaaa ggtattttac agactgtaaa agttgttgat gaattggtca
     3781 aagtaatggg gcggcataag ccagaaaata tcgttattga aatggcacgt gaaaatcaga
     3841 caactcaaaa gggccagaaa aattcgcgag agcgtatgaa acgaatcgaa gaaggtatca
     3901 aagaattagg aagtcagatt cttaaagagc atcctgttga aaatactcaa ttgcaaaatg
     3961 aaaagctcta tctctattat ctccaaaatg gaagagacat gtatgtggac caagaattag
     4021 atattaatcg tttaagtgat tatgatgtcg atcacattgt tccacaaagt ttccttaaag
     4081 acgattcaat agacaataag gtcttaacgc gttctgataa aaatcgtggt aaatgcgata
     4141 acgttccaag tgaagaagta gtcaaaaaga tgaaaaacta ttggagacaa cttctaaacg
     4201 ccaagttaat cactcaacgt aagtttgata atttaacgaa agctgaacgt ggaggtttga
     4261 gtgaacttga taaagctggt tttatcaaac gccaattggt tgaaactcgc caaatcacta
     4321 agcatgtggc acaaattttg gatagtcgca tgaatactaa atacgatgaa aatgataaac
     4381 ttattcgaga ggttaaagtg attaccttaa aatctaaatt agtttctgac ttccgaaaag
     4441 atttccaatt ctataaagta cgtgagatta acaattacca tcatgcccat gatgcgtatc
     4501 taaatgccgt cgttggaact gctttgatta agaaatatcc aaaacttgaa tcggagtttg
     4561 tctatggtga ttataaagtt tatgatgttc gtaaaatgat tgctaagtct gagcaagaaa
     4621 taggcaaagc aaccgcaaaa tatttctttt actctaatat catgaacttc ttcaaaacag
     4681 aaattacact tgcaaatgga gagattcgca aacgccctct aatcgaaact aatggggaaa
     4741 ctggagaaat tgtctgggat aaagggcgag attttgccac agtgcgcaaa gtattgtcca
     4801 tgccccaagt caatattgtc aagaaaacag aagtacagac aggcggattc tccaaggagt
     4861 caattttacc aaaaagaaat tcggacaagc ttattgctcg taaaaaagac tgggatccaa
     4921 aaaaatatgg tggttttgat agtccaacgg tagcttattc agtcctagtg gttgctaagg
     4981 tggaaaaagg gaaatcgaag aagttaaaat ccgttaaaga gttactaggg atcacaatta
     5041 tggaaagaag ttcctttgaa aaaaatccga ttgacttttt agaagctaaa ggatataagg
     5101 aagttaaaaa agacttaatc attaaactac ctaaatatag tctttttgag ttagaaaacg
     5161 gtcgtaaacg gatgctggct agtgccggag aattacaaaa aggaaatgag ctggctctgc
     5221 caagcaaata tgtgaatttt ttatatttag ctagtcatta tgaaaagttg aagggtagtc
     5281 cagaagataa cgaacaaaaa caattgtttg tggagcagca taagcattat ttagatgaga
     5341 ttattgagca aatcagtgaa ttttctaagc gtgttatttt agcagatgcc aatttagata
     5401 aagttcttag tgcatataac aaacatagag acaaaccaat acgtgaacaa gcagaaaata
     5461 ttattcattt atttacgttg acgaatcttg gagctcccgc tgcttttaaa tattttgata
     5521 caacaattga tcgtaaacga tatacgtcta caaaagaagt tttagatgcc actcttatcc
     5581 atcaatccat cactggtctt tatgaaacac gcattgattt gagtcagcta ggaggtgact
     5641 aataacattg gaagtggata acggatccgc gatcgcggcg cgccacctgg tggccggccg
     5701 gtaccacgcg tgcgctctga tccggctgct aacaaagccc gaaaggaagc tgagttggct
     5761 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
     5821 ggttttttgc tgaaaggagg aactatatcc ggatatccac aggacgggtg tggtcgccat
     5881 gatcgcgtag tcgatagtgg ctccaagtag cgaagcgagc aggactgggc ggcggccaaa
     5941 gcggtcggac agtgctccga gaacgggtgc gcatagaaat tgcatcaacg catatagcgc
     6001 tagcagcacg ccatagtgac tggcgatgct gtcggaatgg acgatatccc gcaagaggcc
     6061 cggcagtacc ggcataacca agcctatgcc tacagcatcc agggtgacgg tgccgaggat
     6121 gacgatgagc gcattgttag atttcataca cggtgcctga ctgcgttagc aatttaactg
     6181 tgataaacta ccgcattaaa gcttatcgat gataagctgt caaacatgag aattcttgaa
     6241 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     6301 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     6361 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     6421 aacattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     6481 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     6541 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     6601 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     6661 tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac
     6721 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     6781 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     6841 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     6901 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     6961 acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg
     7021 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     7081 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     7141 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     7201 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     7261 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     7321 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     7381 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     7441 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     7501 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     7561 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     7621 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     7681 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     7741 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     7801 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     7861 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     7921 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     7981 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     8041 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     8101 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     8161 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     8221 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     8281 gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     8341 ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca
     8401 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     8461 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     8521 aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt
     8581 tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag
     8641 cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga
     8701 tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt
     8761 actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg
     8821 atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat
     8881 gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg
     8941 cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat
     9001 gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc
     9061 ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac
     9121 aggagcacga tcatgcgcac ccgtggccag gacccaacgc tgcccgagat gcgccgcgtg
     9181 cggctgctgg agatggcgga cgcgatggat atgttctgcc aagggttggt ttgcgcattc
     9241 acagttctcc gcaagaattg attggctcca attcttggag tggtgaatcc gttagcgagg
     9301 tgccgccggc ttccattcag gtcgaggtgg cccggctcca tgcaccgcga cgcaacgcgg
     9361 ggaggcagac aaggtatagg gcggcgccta caatccatgc caacccgttc catgtgctcg
     9421 ccgaggcggc ataaatcgcc gtgacgatca gcggtccaat gatcgaagtt aggctggtaa
     9481 gagccgcgag cgatccttga agctgtccct gatggtcgtc atctacctgc ctggacagca
     9541 tggcctgcaa cgcgggcatc ccgatgccgc cggaagcgag aagaatcata atggggaagg
     9601 ccatccagcc tcgcgtcgcg aacgccagca agacgtagcc cagcgcgtcg gccgccatgc
     9661 cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt
     9721 gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc
     9781 agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt
     9841 gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga
     9901 aggagctgac tgggttgaag gctctcaagg gcatcggtcg acgctctccc ttatgcgact
     9961 cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga
    10021 atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct gcc
//
