LOCUS       sequence-197535-       10073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for SpCas9 Cluster 1 + Q926A 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: pJSC269 - Bacterial expression plasmid for SpC...
FEATURES             Location/Qualifiers
     source          1..10073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     336..355
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     439..458
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        439..457
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             546..568
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             588..617
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             627..1727
                     /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     1701..1724
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1782..1802
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             join(1809..3881,3882..3884,3885..3887,3888..3893,3894..3899,
                     3900..3902,3903..5915)
                     /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
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRAITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     primer_bind     complement(6041..6059)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6055..6102
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(6468..6473,6474..6490,6491..6496))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(6523..6541)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6609..6713
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6714..6782,6783..7574)
                     /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(6932..6951)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7745..8333
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8234..8253
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8487..8504
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    8519..8658
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(8584..8603)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8744..8766
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(8760..8951)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc
       61 ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctgac tgggttgaag
      121 gctctcaagg gcatcggtcg acgctctccc ttatgcgact cctgcattag gaagcagccc
      181 agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg
      241 gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc
      301 atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca
      361 gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcgag
      421 atctcgatcc cgcgaaatta atacgactca ctatagggag accacaacgg tttccctcta
      481 gtgccggctc cggagagctc tttaattaag cggccgccct gcaggactcg agttctagaa
      541 ataattttgt ttaactttaa gaaggagata tacatatgaa atcttctcac catcaccatc
      601 accatcacca tcaccatggt tcttctatga aaatcgaaga aggtaaactg gtaatctgga
      661 ttaacggcga taaaggctat aacggtctcg ctgaagtcgg taagaaattc gagaaagata
      721 ccggaattaa agtcaccgtt gagcatccgg ataaactgga agagaaattc ccacaggttg
      781 cggcaactgg cgatggccct gacattatct tctgggcaca cgaccgcttt ggtggctacg
      841 ctcaatctgg cctgttggct gaaatcaccc cggacaaagc gttccaggac aagctgtatc
      901 cgtttacctg ggatgccgta cgttacaacg gcaagctgat tgcttacccg atcgctgttg
      961 aagcgttatc gctgatttat aacaaagatc tgctgccgaa cccgccaaaa acctgggaag
     1021 agatcccggc gctggataaa gaactgaaag cgaaaggtaa gagcgcgctg atgttcaacc
     1081 tgcaagaacc gtacttcacc tggccgctga ttgctgctga cgggggttat gcgttcaagt
     1141 atgaaaacgg caagtacgac attaaagacg tgggcgtgga taacgctggc gcgaaagcgg
     1201 gtctgacctt cctggttgac ctgattaaaa acaaacacat gaatgcagac accgattact
     1261 ccatcgcaga agctgccttt aataaaggcg aaacagcgat gaccatcaac ggcccgtggg
     1321 catggtccaa catcgacacc agcaaagtga attatggtgt aacggtactg ccgaccttca
     1381 agggtcaacc atccaaaccg ttcgttggcg tgctgagcgc aggtattaac gccgccagtc
     1441 cgaacaaaga gctggcaaaa gagttcctcg aaaactatct gctgactgat gaaggtctgg
     1501 aagcggttaa taaagacaaa ccgctgggtg ccgtagcgct gaagtcttac gaggaagagt
     1561 tggcgaaaga tccacgtatt gccgccacta tggaaaacgc ccagaaaggt gaaatcatgc
     1621 cgaacatccc gcagatgtcc gctttctggt atgccgtgcg tactgcggtg atcaacgccg
     1681 ccagcggtcg tcagactgtc gatgaagccc tgaaagacgc gcagactaat tcgagctcga
     1741 acaacaacaa caataacaat aacaacaacc tcgggatcga ggaaaacctg tacttccaat
     1801 ccaatgcaat ggataagaaa tactcaatag gcttagatat cggcacaaat agcgtcggat
     1861 gggcggtgat cactgatgaa tataaggttc cgtctaaaaa gttcaaggtt ctgggaaata
     1921 cagaccgcca cagtatcaaa aaaaatctta taggggctct tttatttgac agtggagaga
     1981 cagcggaagc gactcgtctc aaacggacag ctcgtagaag gtatacacgt cggaagaatc
     2041 gtatttgtta tctacaggag attttttcaa atgagatggc gaaagtagat gatagtttct
     2101 ttcatcgact tgaagagtct tttttggtgg aagaagacaa gaagcatgaa cgtcatccta
     2161 tttttggaaa tatagtagat gaagttgctt atcatgagaa atatccaact atctatcatc
     2221 tgcgaaaaaa attggtagat tctactgata aagcggattt gcgcttaatc tatttggcct
     2281 tagcgcatat gattaagttt cgtggtcatt ttttgattga gggagattta aatcctgata
     2341 atagtgatgt ggacaaacta tttatccagt tggtacaaac ctacaatcaa ttatttgaag
     2401 aaaaccctat taacgcaagt ggagtagatg ctaaagcgat tctttctgca cgattgagta
     2461 aatcaagacg attagaaaat ctcattgctc agctccccgg tgagaagaaa aatggcttat
     2521 ttgggaatct cattgctttg tcattgggtt tgacccctaa ttttaaatca aattttgatt
     2581 tggcagaaga tgctaaatta cagctttcaa aagatactta cgatgatgat ttagataatt
     2641 tattggcgca aattggagat caatatgctg atttgttttt ggcagctaag aatttatcag
     2701 atgctatttt actttcagat atcctaagag taaatactga aataactaag gctcccctat
     2761 cagcttcaat gattaaacgc tacgatgaac atcatcaaga cttgactctt ttaaaagctt
     2821 tagttcgaca acaacttcca gaaaagtata aagaaatctt ttttgatcaa tcaaaaaacg
     2881 gatatgcagg ttatattgat gggggagcta gccaagaaga attttataaa tttatcaaac
     2941 caattttaga aaaaatggat ggtactgagg aattattggt gaaactaaat cgtgaagatt
     3001 tgctgcgcaa gcaacggacc tttgacaacg gctctattcc ccatcaaatt cacttgggtg
     3061 agctgcatgc tattttgaga agacaagaag acttttatcc atttttaaaa gacaatcgtg
     3121 agaagattga aaaaatcttg acttttcgaa ttccttatta tgttggtcca ttggcgcgtg
     3181 gcaatagtcg ttttgcatgg atgactcgga agtctgaaga aacaattacc ccatggaatt
     3241 ttgaagaagt tgtcgataaa ggtgcttcag ctcaatcatt tattgaacgc atgacaaact
     3301 ttgataaaaa tcttccaaat gaaaaagtac taccaaaaca tagtttgctt tatgagtatt
     3361 ttacggttta taacgaattg acaaaggtca aatatgttac tgaaggaatg cgaaaaccag
     3421 catttctttc aggtgaacag aagaaagcca ttgttgattt actcttcaaa acaaatcgaa
     3481 aagtaaccgt taagcaatta aaagaagatt atttcaaaaa aatagaatgt tttgatagtg
     3541 ttgaaatttc aggagttgaa gatagattta atgcttcatt aggtacctac catgatttgc
     3601 taaaaattat taaagataaa gattttttgg ataatgaaga aaatgaagat atcttagagg
     3661 atattgtttt aacattgacc ttatttgaag atagggagat gattgaggaa agacttaaaa
     3721 catatgctca cctctttgat gataaggtga tgaaacagct taaacgtcgc cgttatactg
     3781 gttggggacg tttgtctcga aaattgatta atggtattag ggataagcaa tctggcaaaa
     3841 caatattaga ttttttgaaa tcagatggtt ttgccaatcg cgcttttgct gctctgatcg
     3901 ctgatgatag tttgacattt aaagaagaca ttcaaaaagc acaagtgtct ggacaaggcg
     3961 atagtttaca tgaacatatt gcaaatttag ctggtagccc tgctattaaa aaaggtattt
     4021 tacagactgt aaaagttgtt gatgaattgg tcaaagtaat ggggcggcat aagccagaaa
     4081 atatcgttat tgaaatggca cgtgaaaatc agacaactca aaagggccag aaaaattcgc
     4141 gagagcgtat gaaacgaatc gaagaaggta tcaaagaatt aggaagtcag attcttaaag
     4201 agcatcctgt tgaaaatact caattgcaaa atgaaaagct ctatctctat tatctccaaa
     4261 atggaagaga catgtatgtg gaccaagaat tagatattaa tcgtttaagt gattatgatg
     4321 tcgatcacat tgttccacaa agtttcctta aagacgattc aatagacaat aaggtcttaa
     4381 cgcgttctga taaaaatcgt ggtaaatcgg ataacgttcc aagtgaagaa gtagtcaaaa
     4441 agatgaaaaa ctattggaga caacttctaa acgccaagtt aatcactcaa cgtaagtttg
     4501 ataatttaac gaaagctgaa cgtggaggtt tgagtgaact tgataaagct ggttttatca
     4561 aacgccaatt ggttgaaact cgcgctatca ctaagcatgt ggcacaaatt ttggatagtc
     4621 gcatgaatac taaatacgat gaaaatgata aacttattcg agaggttaaa gtgattacct
     4681 taaaatctaa attagtttct gacttccgaa aagatttcca attctataaa gtacgtgaga
     4741 ttaacaatta ccatcatgcc catgatgcgt atctaaatgc cgtcgttgga actgctttga
     4801 ttaagaaata tccaaaactt gaatcggagt ttgtctatgg tgattataaa gtttatgatg
     4861 ttcgtaaaat gattgctaag tctgagcaag aaataggcaa agcaaccgca aaatatttct
     4921 tttactctaa tatcatgaac ttcttcaaaa cagaaattac acttgcaaat ggagagattc
     4981 gcaaacgccc tctaatcgaa actaatgggg aaactggaga aattgtctgg gataaagggc
     5041 gagattttgc cacagtgcgc aaagtattgt ccatgcccca agtcaatatt gtcaagaaaa
     5101 cagaagtaca gacaggcgga ttctccaagg agtcaatttt accaaaaaga aattcggaca
     5161 agcttattgc tcgtaaaaaa gactgggatc caaaaaaata tggtggtttt gatagtccaa
     5221 cggtagctta ttcagtccta gtggttgcta aggtggaaaa agggaaatcg aagaagttaa
     5281 aatccgttaa agagttacta gggatcacaa ttatggaaag aagttccttt gaaaaaaatc
     5341 cgattgactt tttagaagct aaaggatata aggaagttaa aaaagactta atcattaaac
     5401 tacctaaata tagtcttttt gagttagaaa acggtcgtaa acggatgctg gctagtgccg
     5461 gagaattaca aaaaggaaat gagctggctc tgccaagcaa atatgtgaat tttttatatt
     5521 tagctagtca ttatgaaaag ttgaagggta gtccagaaga taacgaacaa aaacaattgt
     5581 ttgtggagca gcataagcat tatttagatg agattattga gcaaatcagt gaattttcta
     5641 agcgtgttat tttagcagat gccaatttag ataaagttct tagtgcatat aacaaacata
     5701 gagacaaacc aatacgtgaa caagcagaaa atattattca tttatttacg ttgacgaatc
     5761 ttggagctcc cgctgctttt aaatattttg atacaacaat tgatcgtaaa cgatatacgt
     5821 ctacaaaaga agttttagat gccactctta tccatcaatc catcactggt ctttatgaaa
     5881 cacgcattga tttgagtcag ctaggaggtg actaataaca ttggaagtgg ataacggatc
     5941 cgcgatcgcg gcgcgccacc tggtggccgg ccggtaccac gcgtgcgcgc tgatccggct
     6001 gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca
     6061 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata
     6121 tccggatatc cacaggacgg gtgtggtcgc catgatcgcg tagtcgatag tggctccaag
     6181 tagcgaagcg agcaggactg ggcggcggcc aaagcggtcg gacagtgctc cgagaacggg
     6241 tgcgcataga aattgcatca acgcatatag cgctagcagc acgccatagt gactggcgat
     6301 gctgtcggaa tggacgatat cccgcaagag gcccggcagt accggcataa ccaagcctat
     6361 gcctacagca tccagggtga cggtgccgag gatgacgatg agcgcattgt tagatttcat
     6421 acacggtgcc tgactgcgtt agcaatttaa ctgtgataaa ctaccgcatt aaagcttatc
     6481 gatgataagc tgtcaaacat gagaattctt gaagacgaaa gggcctcgtg atacgcctat
     6541 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     6601 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     6661 tcatgagaca ataaccctga taaatgcttc aataacattg aaaaaggaag agtatgagta
     6721 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     6781 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     6841 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     6901 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg
     6961 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     7021 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     7081 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     7141 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     7201 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag
     7261 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     7321 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     7381 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     7441 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     7501 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     7561 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     7621 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     7681 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     7741 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     7801 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     7861 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     7921 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     7981 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     8041 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     8101 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     8161 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     8221 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     8281 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     8341 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     8401 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     8461 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     8521 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct
     8581 cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt
     8641 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     8701 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     8761 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg
     8821 tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc
     8881 tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc
     8941 tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg
     9001 atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg
     9061 gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact
     9121 cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag
     9181 catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga
     9241 ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg
     9301 cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg
     9361 caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggc
     9421 caggacccaa cgctgcccga gatgcgccgc gtgcggctgc tggagatggc ggacgcgatg
     9481 gatatgttct gccaagggtt ggtttgcgca ttcacagttc tccgcaagaa ttgattggct
     9541 ccaattcttg gagtggtgaa tccgttagcg aggtgccgcc ggcttccatt caggtcgagg
     9601 tggcccggct ccatgcaccg cgacgcaacg cggggaggca gacaaggtat agggcggcgc
     9661 ctacaatcca tgccaacccg ttccatgtgc tcgccgaggc ggcataaatc gccgtgacga
     9721 tcagcggtcc aatgatcgaa gttaggctgg taagagccgc gagcgatcct tgaagctgtc
     9781 cctgatggtc gtcatctacc tgcctggaca gcatggcctg caacgcgggc atcccgatgc
     9841 cgccggaagc gagaagaatc ataatgggga aggccatcca gcctcgcgtc gcgaacgcca
     9901 gcaagacgta gcccagcgcg tcggccgcca tgccggcgat aatggcctgc ttctcgccga
     9961 aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata
    10021 ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccg
//
