LOCUS       sequence-198035-       10073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for SpCas9 Cluster 1 (HypaCas9) 
            variant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10073)
  AUTHORS   Chen JS, Dagdas YS, Kleinstiver BP, Welch MM, Sousa AA, Harrington 
            LB, Sternberg SH, Joung JK, Yildiz A, Doudna JA
  TITLE     Enhanced proofreading governs CRISPR-Cas9 targeting accuracy.
  JOURNAL   Nature. 2017 Sep 20. doi: 10.1038/nature24268.
  PUBMED    28931002
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"; journalName: "Nature. 
            2017 Sep 20. doi: 10.1038/nature24268."
COMMENT     Sequence Label: pJSC173 - Bacterial expression plasmid for SpC...
FEATURES             Location/Qualifiers
     source          1..10073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     633..652
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     736..755
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        736..754
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             843..865
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             885..914
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             924..2024
                     /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     1998..2021
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             2079..2099
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             join(2106..4178,4179..4181,4182..4184,4185..4190,4191..4196,
                     4197..4199,4200..6212)
                     /codon_start=1
                     /product="Cas9 endonuclease from the Streptococcus pyogenes
                     Type II CRISPR/Cas system, mutated to improve targeting 
                     specificity without compromising on-target activity (Chen 
                     et al., 2017)"
                     /label=HypaCas9
                     /note="carries the mutations N692A, M694A, Q695A, and 
                     H698A"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRAFA
                     ALIADDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     primer_bind     complement(6338..6356)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6352..6399
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(6765..6770,6771..6787,6788..6793))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(6820..6838)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6906..7010
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7011..7079,7080..7871)
                     /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(7229..7248)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8042..8630
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8531..8550
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8784..8801
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    8816..8955
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(8881..8900)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9041..9063
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(9057..9248)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 tgtccctgat ggtcgtcatc tacctgcctg gacagcatgg cctgcaacgc gggcatcccg
       61 atgccgccgg aagcgagaag aatcataatg gggaaggcca tccagcctcg cgtcgcgaac
      121 gccagcaaga cgtagcccag cgcgtcggcc gccatgccgg cgataatggc ctgcttctcg
      181 ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg caagattccg
      241 aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa
      301 atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata
      361 agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct
      421 ctcaagggca tcggtcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt
      481 agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg
      541 cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg
      601 agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca
      661 accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc
      721 tcgatcccgc gaaattaata cgactcacta tagggagacc acaacggttt ccctctagtg
      781 ccggctccgg agagctcttt aattaagcgg ccgccctgca ggactcgagt tctagaaata
      841 attttgttta actttaagaa ggagatatac atatgaaatc ttctcaccat caccatcacc
      901 atcaccatca ccatggttct tctatgaaaa tcgaagaagg taaactggta atctggatta
      961 acggcgataa aggctataac ggtctcgctg aagtcggtaa gaaattcgag aaagataccg
     1021 gaattaaagt caccgttgag catccggata aactggaaga gaaattccca caggttgcgg
     1081 caactggcga tggccctgac attatcttct gggcacacga ccgctttggt ggctacgctc
     1141 aatctggcct gttggctgaa atcaccccgg acaaagcgtt ccaggacaag ctgtatccgt
     1201 ttacctggga tgccgtacgt tacaacggca agctgattgc ttacccgatc gctgttgaag
     1261 cgttatcgct gatttataac aaagatctgc tgccgaaccc gccaaaaacc tgggaagaga
     1321 tcccggcgct ggataaagaa ctgaaagcga aaggtaagag cgcgctgatg ttcaacctgc
     1381 aagaaccgta cttcacctgg ccgctgattg ctgctgacgg gggttatgcg ttcaagtatg
     1441 aaaacggcaa gtacgacatt aaagacgtgg gcgtggataa cgctggcgcg aaagcgggtc
     1501 tgaccttcct ggttgacctg attaaaaaca aacacatgaa tgcagacacc gattactcca
     1561 tcgcagaagc tgcctttaat aaaggcgaaa cagcgatgac catcaacggc ccgtgggcat
     1621 ggtccaacat cgacaccagc aaagtgaatt atggtgtaac ggtactgccg accttcaagg
     1681 gtcaaccatc caaaccgttc gttggcgtgc tgagcgcagg tattaacgcc gccagtccga
     1741 acaaagagct ggcaaaagag ttcctcgaaa actatctgct gactgatgaa ggtctggaag
     1801 cggttaataa agacaaaccg ctgggtgccg tagcgctgaa gtcttacgag gaagagttgg
     1861 cgaaagatcc acgtattgcc gccactatgg aaaacgccca gaaaggtgaa atcatgccga
     1921 acatcccgca gatgtccgct ttctggtatg ccgtgcgtac tgcggtgatc aacgccgcca
     1981 gcggtcgtca gactgtcgat gaagccctga aagacgcgca gactaattcg agctcgaaca
     2041 acaacaacaa taacaataac aacaacctcg ggatcgagga aaacctgtac ttccaatcca
     2101 atgcaatgga taagaaatac tcaataggct tagatatcgg cacaaatagc gtcggatggg
     2161 cggtgatcac tgatgaatat aaggttccgt ctaaaaagtt caaggttctg ggaaatacag
     2221 accgccacag tatcaaaaaa aatcttatag gggctctttt atttgacagt ggagagacag
     2281 cggaagcgac tcgtctcaaa cggacagctc gtagaaggta tacacgtcgg aagaatcgta
     2341 tttgttatct acaggagatt ttttcaaatg agatggcgaa agtagatgat agtttctttc
     2401 atcgacttga agagtctttt ttggtggaag aagacaagaa gcatgaacgt catcctattt
     2461 ttggaaatat agtagatgaa gttgcttatc atgagaaata tccaactatc tatcatctgc
     2521 gaaaaaaatt ggtagattct actgataaag cggatttgcg cttaatctat ttggccttag
     2581 cgcatatgat taagtttcgt ggtcattttt tgattgaggg agatttaaat cctgataata
     2641 gtgatgtgga caaactattt atccagttgg tacaaaccta caatcaatta tttgaagaaa
     2701 accctattaa cgcaagtgga gtagatgcta aagcgattct ttctgcacga ttgagtaaat
     2761 caagacgatt agaaaatctc attgctcagc tccccggtga gaagaaaaat ggcttatttg
     2821 ggaatctcat tgctttgtca ttgggtttga cccctaattt taaatcaaat tttgatttgg
     2881 cagaagatgc taaattacag ctttcaaaag atacttacga tgatgattta gataatttat
     2941 tggcgcaaat tggagatcaa tatgctgatt tgtttttggc agctaagaat ttatcagatg
     3001 ctattttact ttcagatatc ctaagagtaa atactgaaat aactaaggct cccctatcag
     3061 cttcaatgat taaacgctac gatgaacatc atcaagactt gactctttta aaagctttag
     3121 ttcgacaaca acttccagaa aagtataaag aaatcttttt tgatcaatca aaaaacggat
     3181 atgcaggtta tattgatggg ggagctagcc aagaagaatt ttataaattt atcaaaccaa
     3241 ttttagaaaa aatggatggt actgaggaat tattggtgaa actaaatcgt gaagatttgc
     3301 tgcgcaagca acggaccttt gacaacggct ctattcccca tcaaattcac ttgggtgagc
     3361 tgcatgctat tttgagaaga caagaagact tttatccatt tttaaaagac aatcgtgaga
     3421 agattgaaaa aatcttgact tttcgaattc cttattatgt tggtccattg gcgcgtggca
     3481 atagtcgttt tgcatggatg actcggaagt ctgaagaaac aattacccca tggaattttg
     3541 aagaagttgt cgataaaggt gcttcagctc aatcatttat tgaacgcatg acaaactttg
     3601 ataaaaatct tccaaatgaa aaagtactac caaaacatag tttgctttat gagtatttta
     3661 cggtttataa cgaattgaca aaggtcaaat atgttactga aggaatgcga aaaccagcat
     3721 ttctttcagg tgaacagaag aaagccattg ttgatttact cttcaaaaca aatcgaaaag
     3781 taaccgttaa gcaattaaaa gaagattatt tcaaaaaaat agaatgtttt gatagtgttg
     3841 aaatttcagg agttgaagat agatttaatg cttcattagg tacctaccat gatttgctaa
     3901 aaattattaa agataaagat tttttggata atgaagaaaa tgaagatatc ttagaggata
     3961 ttgttttaac attgacctta tttgaagata gggagatgat tgaggaaaga cttaaaacat
     4021 atgctcacct ctttgatgat aaggtgatga aacagcttaa acgtcgccgt tatactggtt
     4081 ggggacgttt gtctcgaaaa ttgattaatg gtattaggga taagcaatct ggcaaaacaa
     4141 tattagattt tttgaaatca gatggttttg ccaatcgcgc ttttgctgct ctgatcgctg
     4201 atgatagttt gacatttaaa gaagacattc aaaaagcaca agtgtctgga caaggcgata
     4261 gtttacatga acatattgca aatttagctg gtagccctgc tattaaaaaa ggtattttac
     4321 agactgtaaa agttgttgat gaattggtca aagtaatggg gcggcataag ccagaaaata
     4381 tcgttattga aatggcacgt gaaaatcaga caactcaaaa gggccagaaa aattcgcgag
     4441 agcgtatgaa acgaatcgaa gaaggtatca aagaattagg aagtcagatt cttaaagagc
     4501 atcctgttga aaatactcaa ttgcaaaatg aaaagctcta tctctattat ctccaaaatg
     4561 gaagagacat gtatgtggac caagaattag atattaatcg tttaagtgat tatgatgtcg
     4621 atcacattgt tccacaaagt ttccttaaag acgattcaat agacaataag gtcttaacgc
     4681 gttctgataa aaatcgtggt aaatcggata acgttccaag tgaagaagta gtcaaaaaga
     4741 tgaaaaacta ttggagacaa cttctaaacg ccaagttaat cactcaacgt aagtttgata
     4801 atttaacgaa agctgaacgt ggaggtttga gtgaacttga taaagctggt tttatcaaac
     4861 gccaattggt tgaaactcgc caaatcacta agcatgtggc acaaattttg gatagtcgca
     4921 tgaatactaa atacgatgaa aatgataaac ttattcgaga ggttaaagtg attaccttaa
     4981 aatctaaatt agtttctgac ttccgaaaag atttccaatt ctataaagta cgtgagatta
     5041 acaattacca tcatgcccat gatgcgtatc taaatgccgt cgttggaact gctttgatta
     5101 agaaatatcc aaaacttgaa tcggagtttg tctatggtga ttataaagtt tatgatgttc
     5161 gtaaaatgat tgctaagtct gagcaagaaa taggcaaagc aaccgcaaaa tatttctttt
     5221 actctaatat catgaacttc ttcaaaacag aaattacact tgcaaatgga gagattcgca
     5281 aacgccctct aatcgaaact aatggggaaa ctggagaaat tgtctgggat aaagggcgag
     5341 attttgccac agtgcgcaaa gtattgtcca tgccccaagt caatattgtc aagaaaacag
     5401 aagtacagac aggcggattc tccaaggagt caattttacc aaaaagaaat tcggacaagc
     5461 ttattgctcg taaaaaagac tgggatccaa aaaaatatgg tggttttgat agtccaacgg
     5521 tagcttattc agtcctagtg gttgctaagg tggaaaaagg gaaatcgaag aagttaaaat
     5581 ccgttaaaga gttactaggg atcacaatta tggaaagaag ttcctttgaa aaaaatccga
     5641 ttgacttttt agaagctaaa ggatataagg aagttaaaaa agacttaatc attaaactac
     5701 ctaaatatag tctttttgag ttagaaaacg gtcgtaaacg gatgctggct agtgccggag
     5761 aattacaaaa aggaaatgag ctggctctgc caagcaaata tgtgaatttt ttatatttag
     5821 ctagtcatta tgaaaagttg aagggtagtc cagaagataa cgaacaaaaa caattgtttg
     5881 tggagcagca taagcattat ttagatgaga ttattgagca aatcagtgaa ttttctaagc
     5941 gtgttatttt agcagatgcc aatttagata aagttcttag tgcatataac aaacatagag
     6001 acaaaccaat acgtgaacaa gcagaaaata ttattcattt atttacgttg acgaatcttg
     6061 gagctcccgc tgcttttaaa tattttgata caacaattga tcgtaaacga tatacgtcta
     6121 caaaagaagt tttagatgcc actcttatcc atcaatccat cactggtctt tatgaaacac
     6181 gcattgattt gagtcagcta ggaggtgact aataacattg gaagtggata acggatccgc
     6241 gatcgcggcg cgccacctgg tggccggccg gtaccacgcg tgcgcgctga tccggctgct
     6301 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     6361 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
     6421 ggatatccac aggacgggtg tggtcgccat gatcgcgtag tcgatagtgg ctccaagtag
     6481 cgaagcgagc aggactgggc ggcggccaaa gcggtcggac agtgctccga gaacgggtgc
     6541 gcatagaaat tgcatcaacg catatagcgc tagcagcacg ccatagtgac tggcgatgct
     6601 gtcggaatgg acgatatccc gcaagaggcc cggcagtacc ggcataacca agcctatgcc
     6661 tacagcatcc agggtgacgg tgccgaggat gacgatgagc gcattgttag atttcataca
     6721 cggtgcctga ctgcgttagc aatttaactg tgataaacta ccgcattaaa gcttatcgat
     6781 gataagctgt caaacatgag aattcttgaa gacgaaaggg cctcgtgata cgcctatttt
     6841 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     6901 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     6961 tgagacaata accctgataa atgcttcaat aacattgaaa aaggaagagt atgagtattc
     7021 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     7081 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     7141 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     7201 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg
     7261 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     7321 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     7381 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     7441 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     7501 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa
     7561 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     7621 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     7681 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     7741 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     7801 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     7861 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     7921 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     7981 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     8041 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     8101 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     8161 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     8221 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     8281 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     8341 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     8401 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     8461 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     8521 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     8581 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     8641 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     8701 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     8761 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     8821 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag
     8881 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     8941 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     9001 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     9061 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg
     9121 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
     9181 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
     9241 ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg
     9301 aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa
     9361 cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag
     9421 ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat
     9481 cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt
     9541 tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag
     9601 cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa
     9661 ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggccag
     9721 gacccaacgc tgcccgagat gcgccgcgtg cggctgctgg agatggcgga cgcgatggat
     9781 atgttctgcc aagggttggt ttgcgcattc acagttctcc gcaagaattg attggctcca
     9841 attcttggag tggtgaatcc gttagcgagg tgccgccggc ttccattcag gtcgaggtgg
     9901 cccggctcca tgcaccgcga cgcaacgcgg ggaggcagac aaggtatagg gcggcgccta
     9961 caatccatgc caacccgttc catgtgctcg ccgaggcggc ataaatcgcc gtgacgatca
    10021 gcggtccaat gatcgaagtt aggctggtaa gagccgcgag cgatccttga agc
//
