LOCUS       sequence-198112-       10073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for SpCas9 + 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: pJSC176 - Bacterial expression plasmid for SpC...
FEATURES             Location/Qualifiers
     source          1..10073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     146..165
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     249..268
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        249..267
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             356..378
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             398..427
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             437..1537
                     /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     1511..1534
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1592..1612
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1619..5725
                     /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
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRAITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     primer_bind     complement(5851..5869)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5865..5912
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(6278..6283,6284..6300,6301..6306))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(6333..6351)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6419..6523
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6524..6592,6593..7384)
                     /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(6742..6761)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7555..8143
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8044..8063
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8297..8314
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    8329..8468
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(8394..8413)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8554..8576
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(8570..8761)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca
       61 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
      121 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
      181 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atctcgatcc
      241 cgcgaaatta atacgactca ctatagggag accacaacgg tttccctcta gtgccggctc
      301 cggagagctc tttaattaag cggccgccct gcaggactcg agttctagaa ataattttgt
      361 ttaactttaa gaaggagata tacatatgaa atcttctcac catcaccatc accatcacca
      421 tcaccatggt tcttctatga aaatcgaaga aggtaaactg gtaatctgga ttaacggcga
      481 taaaggctat aacggtctcg ctgaagtcgg taagaaattc gagaaagata ccggaattaa
      541 agtcaccgtt gagcatccgg ataaactgga agagaaattc ccacaggttg cggcaactgg
      601 cgatggccct gacattatct tctgggcaca cgaccgcttt ggtggctacg ctcaatctgg
      661 cctgttggct gaaatcaccc cggacaaagc gttccaggac aagctgtatc cgtttacctg
      721 ggatgccgta cgttacaacg gcaagctgat tgcttacccg atcgctgttg aagcgttatc
      781 gctgatttat aacaaagatc tgctgccgaa cccgccaaaa acctgggaag agatcccggc
      841 gctggataaa gaactgaaag cgaaaggtaa gagcgcgctg atgttcaacc tgcaagaacc
      901 gtacttcacc tggccgctga ttgctgctga cgggggttat gcgttcaagt atgaaaacgg
      961 caagtacgac attaaagacg tgggcgtgga taacgctggc gcgaaagcgg gtctgacctt
     1021 cctggttgac ctgattaaaa acaaacacat gaatgcagac accgattact ccatcgcaga
     1081 agctgccttt aataaaggcg aaacagcgat gaccatcaac ggcccgtggg catggtccaa
     1141 catcgacacc agcaaagtga attatggtgt aacggtactg ccgaccttca agggtcaacc
     1201 atccaaaccg ttcgttggcg tgctgagcgc aggtattaac gccgccagtc cgaacaaaga
     1261 gctggcaaaa gagttcctcg aaaactatct gctgactgat gaaggtctgg aagcggttaa
     1321 taaagacaaa ccgctgggtg ccgtagcgct gaagtcttac gaggaagagt tggcgaaaga
     1381 tccacgtatt gccgccacta tggaaaacgc ccagaaaggt gaaatcatgc cgaacatccc
     1441 gcagatgtcc gctttctggt atgccgtgcg tactgcggtg atcaacgccg ccagcggtcg
     1501 tcagactgtc gatgaagccc tgaaagacgc gcagactaat tcgagctcga acaacaacaa
     1561 caataacaat aacaacaacc tcgggatcga ggaaaacctg tacttccaat ccaatgcaat
     1621 ggataagaaa tactcaatag gcttagatat cggcacaaat agcgtcggat gggcggtgat
     1681 cactgatgaa tataaggttc cgtctaaaaa gttcaaggtt ctgggaaata cagaccgcca
     1741 cagtatcaaa aaaaatctta taggggctct tttatttgac agtggagaga cagcggaagc
     1801 gactcgtctc aaacggacag ctcgtagaag gtatacacgt cggaagaatc gtatttgtta
     1861 tctacaggag attttttcaa atgagatggc gaaagtagat gatagtttct ttcatcgact
     1921 tgaagagtct tttttggtgg aagaagacaa gaagcatgaa cgtcatccta tttttggaaa
     1981 tatagtagat gaagttgctt atcatgagaa atatccaact atctatcatc tgcgaaaaaa
     2041 attggtagat tctactgata aagcggattt gcgcttaatc tatttggcct tagcgcatat
     2101 gattaagttt cgtggtcatt ttttgattga gggagattta aatcctgata atagtgatgt
     2161 ggacaaacta tttatccagt tggtacaaac ctacaatcaa ttatttgaag aaaaccctat
     2221 taacgcaagt ggagtagatg ctaaagcgat tctttctgca cgattgagta aatcaagacg
     2281 attagaaaat ctcattgctc agctccccgg tgagaagaaa aatggcttat ttgggaatct
     2341 cattgctttg tcattgggtt tgacccctaa ttttaaatca aattttgatt tggcagaaga
     2401 tgctaaatta cagctttcaa aagatactta cgatgatgat ttagataatt tattggcgca
     2461 aattggagat caatatgctg atttgttttt ggcagctaag aatttatcag atgctatttt
     2521 actttcagat atcctaagag taaatactga aataactaag gctcccctat cagcttcaat
     2581 gattaaacgc tacgatgaac atcatcaaga cttgactctt ttaaaagctt tagttcgaca
     2641 acaacttcca gaaaagtata aagaaatctt ttttgatcaa tcaaaaaacg gatatgcagg
     2701 ttatattgat gggggagcta gccaagaaga attttataaa tttatcaaac caattttaga
     2761 aaaaatggat ggtactgagg aattattggt gaaactaaat cgtgaagatt tgctgcgcaa
     2821 gcaacggacc tttgacaacg gctctattcc ccatcaaatt cacttgggtg agctgcatgc
     2881 tattttgaga agacaagaag acttttatcc atttttaaaa gacaatcgtg agaagattga
     2941 aaaaatcttg acttttcgaa ttccttatta tgttggtcca ttggcgcgtg gcaatagtcg
     3001 ttttgcatgg atgactcgga agtctgaaga aacaattacc ccatggaatt ttgaagaagt
     3061 tgtcgataaa ggtgcttcag ctcaatcatt tattgaacgc atgacaaact ttgataaaaa
     3121 tcttccaaat gaaaaagtac taccaaaaca tagtttgctt tatgagtatt ttacggttta
     3181 taacgaattg acaaaggtca aatatgttac tgaaggaatg cgaaaaccag catttctttc
     3241 aggtgaacag aagaaagcca ttgttgattt actcttcaaa acaaatcgaa aagtaaccgt
     3301 taagcaatta aaagaagatt atttcaaaaa aatagaatgt tttgatagtg ttgaaatttc
     3361 aggagttgaa gatagattta atgcttcatt aggtacctac catgatttgc taaaaattat
     3421 taaagataaa gattttttgg ataatgaaga aaatgaagat atcttagagg atattgtttt
     3481 aacattgacc ttatttgaag atagggagat gattgaggaa agacttaaaa catatgctca
     3541 cctctttgat gataaggtga tgaaacagct taaacgtcgc cgttatactg gttggggacg
     3601 tttgtctcga aaattgatta atggtattag ggataagcaa tctggcaaaa caatattaga
     3661 ttttttgaaa tcagatggtt ttgccaatcg caattttatg cagctgatcc atgatgatag
     3721 tttgacattt aaagaagaca ttcaaaaagc acaagtgtct ggacaaggcg atagtttaca
     3781 tgaacatatt gcaaatttag ctggtagccc tgctattaaa aaaggtattt tacagactgt
     3841 aaaagttgtt gatgaattgg tcaaagtaat ggggcggcat aagccagaaa atatcgttat
     3901 tgaaatggca cgtgaaaatc agacaactca aaagggccag aaaaattcgc gagagcgtat
     3961 gaaacgaatc gaagaaggta tcaaagaatt aggaagtcag attcttaaag agcatcctgt
     4021 tgaaaatact caattgcaaa atgaaaagct ctatctctat tatctccaaa atggaagaga
     4081 catgtatgtg gaccaagaat tagatattaa tcgtttaagt gattatgatg tcgatcacat
     4141 tgttccacaa agtttcctta aagacgattc aatagacaat aaggtcttaa cgcgttctga
     4201 taaaaatcgt ggtaaatcgg ataacgttcc aagtgaagaa gtagtcaaaa agatgaaaaa
     4261 ctattggaga caacttctaa acgccaagtt aatcactcaa cgtaagtttg ataatttaac
     4321 gaaagctgaa cgtggaggtt tgagtgaact tgataaagct ggttttatca aacgccaatt
     4381 ggttgaaact cgcgctatca ctaagcatgt ggcacaaatt ttggatagtc gcatgaatac
     4441 taaatacgat gaaaatgata aacttattcg agaggttaaa gtgattacct taaaatctaa
     4501 attagtttct gacttccgaa aagatttcca attctataaa gtacgtgaga ttaacaatta
     4561 ccatcatgcc catgatgcgt atctaaatgc cgtcgttgga actgctttga ttaagaaata
     4621 tccaaaactt gaatcggagt ttgtctatgg tgattataaa gtttatgatg ttcgtaaaat
     4681 gattgctaag tctgagcaag aaataggcaa agcaaccgca aaatatttct tttactctaa
     4741 tatcatgaac ttcttcaaaa cagaaattac acttgcaaat ggagagattc gcaaacgccc
     4801 tctaatcgaa actaatgggg aaactggaga aattgtctgg gataaagggc gagattttgc
     4861 cacagtgcgc aaagtattgt ccatgcccca agtcaatatt gtcaagaaaa cagaagtaca
     4921 gacaggcgga ttctccaagg agtcaatttt accaaaaaga aattcggaca agcttattgc
     4981 tcgtaaaaaa gactgggatc caaaaaaata tggtggtttt gatagtccaa cggtagctta
     5041 ttcagtccta gtggttgcta aggtggaaaa agggaaatcg aagaagttaa aatccgttaa
     5101 agagttacta gggatcacaa ttatggaaag aagttccttt gaaaaaaatc cgattgactt
     5161 tttagaagct aaaggatata aggaagttaa aaaagactta atcattaaac tacctaaata
     5221 tagtcttttt gagttagaaa acggtcgtaa acggatgctg gctagtgccg gagaattaca
     5281 aaaaggaaat gagctggctc tgccaagcaa atatgtgaat tttttatatt tagctagtca
     5341 ttatgaaaag ttgaagggta gtccagaaga taacgaacaa aaacaattgt ttgtggagca
     5401 gcataagcat tatttagatg agattattga gcaaatcagt gaattttcta agcgtgttat
     5461 tttagcagat gccaatttag ataaagttct tagtgcatat aacaaacata gagacaaacc
     5521 aatacgtgaa caagcagaaa atattattca tttatttacg ttgacgaatc ttggagctcc
     5581 cgctgctttt aaatattttg atacaacaat tgatcgtaaa cgatatacgt ctacaaaaga
     5641 agttttagat gccactctta tccatcaatc catcactggt ctttatgaaa cacgcattga
     5701 tttgagtcag ctaggaggtg actaataaca ttggaagtgg ataacggatc cgcgatcgcg
     5761 gcgcgccacc tggtggccgg ccggtaccac gcgtgcgcgc tgatccggct gctaacaaag
     5821 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
     5881 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggatatc
     5941 cacaggacgg gtgtggtcgc catgatcgcg tagtcgatag tggctccaag tagcgaagcg
     6001 agcaggactg ggcggcggcc aaagcggtcg gacagtgctc cgagaacggg tgcgcataga
     6061 aattgcatca acgcatatag cgctagcagc acgccatagt gactggcgat gctgtcggaa
     6121 tggacgatat cccgcaagag gcccggcagt accggcataa ccaagcctat gcctacagca
     6181 tccagggtga cggtgccgag gatgacgatg agcgcattgt tagatttcat acacggtgcc
     6241 tgactgcgtt agcaatttaa ctgtgataaa ctaccgcatt aaagcttatc gatgataagc
     6301 tgtcaaacat gagaattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
     6361 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     6421 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     6481 ataaccctga taaatgcttc aataacattg aaaaaggaag agtatgagta ttcaacattt
     6541 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     6601 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     6661 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     6721 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
     6781 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     6841 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     6901 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     6961 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     7021 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
     7081 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     7141 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     7201 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     7261 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     7321 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     7381 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     7441 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     7501 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     7561 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     7621 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     7681 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     7741 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     7801 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     7861 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     7921 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     7981 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     8041 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     8101 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     8161 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     8221 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     8281 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta
     8341 ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc
     8401 tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca
     8461 tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     8521 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     8581 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa
     8641 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg
     8701 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca
     8761 ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag
     8821 agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg
     8881 agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat
     8941 gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga
     9001 tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa
     9061 cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt
     9121 cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc
     9181 agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggc caggacccaa
     9241 cgctgcccga gatgcgccgc gtgcggctgc tggagatggc ggacgcgatg gatatgttct
     9301 gccaagggtt ggtttgcgca ttcacagttc tccgcaagaa ttgattggct ccaattcttg
     9361 gagtggtgaa tccgttagcg aggtgccgcc ggcttccatt caggtcgagg tggcccggct
     9421 ccatgcaccg cgacgcaacg cggggaggca gacaaggtat agggcggcgc ctacaatcca
     9481 tgccaacccg ttccatgtgc tcgccgaggc ggcataaatc gccgtgacga tcagcggtcc
     9541 aatgatcgaa gttaggctgg taagagccgc gagcgatcct tgaagctgtc cctgatggtc
     9601 gtcatctacc tgcctggaca gcatggcctg caacgcgggc atcccgatgc cgccggaagc
     9661 gagaagaatc ataatgggga aggccatcca gcctcgcgtc gcgaacgcca gcaagacgta
     9721 gcccagcgcg tcggccgcca tgccggcgat aatggcctgc ttctcgccga aacgtttggt
     9781 ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata ccgcaagcga
     9841 caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc
     9901 tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat
     9961 agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg
    10021 tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta ggt
//
