LOCUS       sequence-198110-       10073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for SpCas9 + N497A/R661A/Q695A, HNH 
            FRET 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: pJSC120 - Bacterial expression plasmid for SpC...
FEATURES             Location/Qualifiers
     source          1..10073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     133..152
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     236..255
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        236..254
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             343..365
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             385..414
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             424..1524
                     /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     1498..1521
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1579..1599
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1606..5712
                     /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
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTAFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIESFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGALSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     ALIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKCDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     primer_bind     complement(5838..5856)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5852..5899
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(6265..6270,6271..6287,6288..6293))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(6320..6338)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6406..6510
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6511..6579,6580..7371)
                     /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(6729..6748)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7542..8130
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8031..8050
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8284..8301
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    8316..8455
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(8381..8400)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8541..8563
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(8557..8748)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac
       61 ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg
      121 atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt
      181 gatgccggcc acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc gaaattaata
      241 cgactcacta tagggagacc acaacggttt ccctctagtg ccggctccgg agagctcttt
      301 aattaagcgg ccgccctgca ggactcgagt tctagaaata attttgttta actttaagaa
      361 ggagatatac atatgaaatc ttctcaccat caccatcacc atcaccatca ccatggttct
      421 tctatgaaaa tcgaagaagg taaactggta atctggatta acggcgataa aggctataac
      481 ggtctcgctg aagtcggtaa gaaattcgag aaagataccg gaattaaagt caccgttgag
      541 catccggata aactggaaga gaaattccca caggttgcgg caactggcga tggccctgac
      601 attatcttct gggcacacga ccgctttggt ggctacgctc aatctggcct gttggctgaa
      661 atcaccccgg acaaagcgtt ccaggacaag ctgtatccgt ttacctggga tgccgtacgt
      721 tacaacggca agctgattgc ttacccgatc gctgttgaag cgttatcgct gatttataac
      781 aaagatctgc tgccgaaccc gccaaaaacc tgggaagaga tcccggcgct ggataaagaa
      841 ctgaaagcga aaggtaagag cgcgctgatg ttcaacctgc aagaaccgta cttcacctgg
      901 ccgctgattg ctgctgacgg gggttatgcg ttcaagtatg aaaacggcaa gtacgacatt
      961 aaagacgtgg gcgtggataa cgctggcgca aaagcgggtc tgaccttcct ggttgacctg
     1021 attaaaaaca aacacatgaa tgcagacacc gattactcca tcgcagaagc tgcctttaat
     1081 aaaggcgaaa cagcgatgac catcaacggc ccgtgggcat ggtccaacat cgacaccagc
     1141 aaagtgaatt atggtgtaac ggtactgccg accttcaagg gtcaaccatc caaaccgttc
     1201 gttggcgtgc tgagcgcagg tattaacgcc gccagtccga acaaagagct ggcaaaagag
     1261 ttcctcgaaa actatctgct gactgatgaa ggtctggaag cggttaataa agacaaaccg
     1321 ctgggtgccg tagcgctgaa gtcttacgag gaagagttgg cgaaagatcc acgtattgcc
     1381 gccactatgg aaaacgccca gaaaggtgaa atcatgccga acatcccgca gatgtccgct
     1441 ttctggtatg ccgtgcgtac tgcggtgatc aacgccgcca gcggtcgtca gactgtcgat
     1501 gaagccctga aagacgcgca gactaattcg agctcgaaca acaacaacaa taacaataac
     1561 aacaacctcg ggatcgagga aaacctgtac ttccaatcca atgcaatgga taagaaatac
     1621 tcaataggct tagatatcgg cacaaatagc gtcggatggg cggtgatcac tgatgaatat
     1681 aaggttccgt ctaaaaagtt caaggttctg ggaaatacag accgccacag tatcaaaaaa
     1741 aatcttatag gggctctttt atttgacagt ggagagacag cggaagcgac tcgtctcaaa
     1801 cggacagctc gtagaaggta tacacgtcgg aagaatcgta tttcttatct acaggagatt
     1861 ttttcaaatg agatggcgaa agtagatgat agtttctttc atcgacttga agagtctttt
     1921 ttggtggaag aagacaagaa gcatgaacgt catcctattt ttggaaatat agtagatgaa
     1981 gttgcttatc atgagaaata tccaactatc tatcatctgc gaaaaaaatt ggtagattct
     2041 actgataaag cggatttgcg cttaatctat ttggccttag cgcatatgat taagtttcgt
     2101 ggtcattttt tgattgaggg agatttaaat cctgataata gtgatgtgga caaactattt
     2161 atccagttgg tacaaaccta caatcaatta tttgaagaaa accctattaa cgcaagtgga
     2221 gtagatgcta aagcgattct ttctgcacga ttgagtaaat caagacgatt agaaaatctc
     2281 attgctcagc tccccggtga gaagaaaaat ggcttatttg ggaatctcat tgctttgtca
     2341 ttgggtttga cccctaattt taaatcaaat tttgatttgg cagaagatgc taaattacag
     2401 ctttcaaaag atacttacga tgatgattta gataatttat tggcgcaaat tggagatcaa
     2461 tatgctgatt tgtttttggc agctaagaat ttatcagatg ctattttact ttcagatatc
     2521 ctaagagtaa atactgaaat aactaaggct cccctatcag cttcaatgat taaacgctac
     2581 gatgaacatc atcaagactt gactctttta aaagctttag ttcgacaaca acttccagaa
     2641 aagtataaag aaatcttttt tgatcaatgt aaaaacggat atgcaggtta tattgatggg
     2701 ggagctagcc aagaagaatt ttataaattt atcaaaccaa ttttagaaaa aatggatggt
     2761 actgaggaat tattggtgaa actaaatcgt gaagatttgc tgcgcaagca acggaccttt
     2821 gacaacggct ctattcccca tcaaattcac ttgggtgagc tgcatgctat tttgagaaga
     2881 caagaagact tttatccatt tttaaaagac aatcgtgaga agattgaaaa aatcttgact
     2941 tttcgaattc cttattatgt tggtccattg gcgcgtggca atagtcgttt tgcatggatg
     3001 actcggaagt ctgaagaaac aattacccca tggaattttg aagaagttgt cgataaaggt
     3061 gcttcagctc aatcatttat tgaacgcatg acagcgtttg ataaaaatct tccaaatgaa
     3121 aaagtactac caaaacatag tttgctttat gagtatttta cggtttataa cgaattgaca
     3181 aaggtcaaat atgttactga aggaatgcga aaaccagcat ttctttcagg tgaacagaag
     3241 aaagccattg ttgatttact cttcaaaaca aatcgaaaag taaccgttaa gcaattaaaa
     3301 gaagattatt tcaaaaaaat agaatctttt gatagtgttg aaatttcagg agttgaagat
     3361 agatttaatg cttcattagg tacctaccat gatttgctaa aaattattaa agataaagat
     3421 tttttggata atgaagaaaa tgaagatatc ttagaggata ttgttttaac attgacctta
     3481 tttgaagata gggagatgat tgaggaaaga cttaaaacat atgctcacct ctttgatgat
     3541 aaggtgatga aacagcttaa acgtcgccgt tatactggtt ggggagcgtt gtctcgaaaa
     3601 ttgattaatg gtattaggga taagcaatct ggcaaaacaa tattagattt tttgaaatca
     3661 gatggttttg ccaatcgcaa ttttatggcc ctgatccatg atgatagttt gacatttaaa
     3721 gaagacattc aaaaagcaca agtgtctgga caaggcgata gtttacatga acatattgca
     3781 aatttagctg gtagccctgc tattaaaaaa ggtattttac agactgtaaa agttgttgat
     3841 gaattggtca aagtaatggg gcggcataag ccagaaaata tcgttattga aatggcacgt
     3901 gaaaatcaga caactcaaaa gggccagaaa aattcgcgag agcgtatgaa acgaatcgaa
     3961 gaaggtatca aagaattagg aagtcagatt cttaaagagc atcctgttga aaatactcaa
     4021 ttgcaaaatg aaaagctcta tctctattat ctccaaaatg gaagagacat gtatgtggac
     4081 caagaattag atattaatcg tttaagtgat tatgatgtcg atcacattgt tccacaaagt
     4141 ttccttaaag acgattcaat agacaataag gtcttaacgc gttctgataa aaatcgtggt
     4201 aaatgcgata acgttccaag tgaagaagta gtcaaaaaga tgaaaaacta ttggagacaa
     4261 cttctaaacg ccaagttaat cactcaacgt aagtttgata atttaacgaa agctgaacgt
     4321 ggaggtttga gtgaacttga taaagctggt tttatcaaac gccaattggt tgaaactcgc
     4381 caaatcacta agcatgtggc acaaattttg gatagtcgca tgaatactaa atacgatgaa
     4441 aatgataaac ttattcgaga ggttaaagtg attaccttaa aatctaaatt agtttctgac
     4501 ttccgaaaag atttccaatt ctataaagta cgtgagatta acaattacca tcatgcccat
     4561 gatgcgtatc taaatgccgt cgttggaact gctttgatta agaaatatcc aaaacttgaa
     4621 tcggagtttg tctatggtga ttataaagtt tatgatgttc gtaaaatgat tgctaagtct
     4681 gagcaagaaa taggcaaagc aaccgcaaaa tatttctttt actctaatat catgaacttc
     4741 ttcaaaacag aaattacact tgcaaatgga gagattcgca aacgccctct aatcgaaact
     4801 aatggggaaa ctggagaaat tgtctgggat aaagggcgag attttgccac agtgcgcaaa
     4861 gtattgtcca tgccccaagt caatattgtc aagaaaacag aagtacagac aggcggattc
     4921 tccaaggagt caattttacc aaaaagaaat tcggacaagc ttattgctcg taaaaaagac
     4981 tgggatccaa aaaaatatgg tggttttgat agtccaacgg tagcttattc agtcctagtg
     5041 gttgctaagg tggaaaaagg gaaatcgaag aagttaaaat ccgttaaaga gttactaggg
     5101 atcacaatta tggaaagaag ttcctttgaa aaaaatccga ttgacttttt agaagctaaa
     5161 ggatataagg aagttaaaaa agacttaatc attaaactac ctaaatatag tctttttgag
     5221 ttagaaaacg gtcgtaaacg gatgctggct agtgccggag aattacaaaa aggaaatgag
     5281 ctggctctgc caagcaaata tgtgaatttt ttatatttag ctagtcatta tgaaaagttg
     5341 aagggtagtc cagaagataa cgaacaaaaa caattgtttg tggagcagca taagcattat
     5401 ttagatgaga ttattgagca aatcagtgaa ttttctaagc gtgttatttt agcagatgcc
     5461 aatttagata aagttcttag tgcatataac aaacatagag acaaaccaat acgtgaacaa
     5521 gcagaaaata ttattcattt atttacgttg acgaatcttg gagctcccgc tgcttttaaa
     5581 tattttgata caacaattga tcgtaaacga tatacgtcta caaaagaagt tttagatgcc
     5641 actcttatcc atcaatccat cactggtctt tatgaaacac gcattgattt gagtcagcta
     5701 ggaggtgact aataacattg gaagtggata acggatccgc gatcgcggcg cgccacctgg
     5761 tggccggccg gtaccacgcg tgcgctctga tccggctgct aacaaagccc gaaaggaagc
     5821 tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
     5881 ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggatatccac aggacgggtg
     5941 tggtcgccat gatcgcgtag tcgatagtgg ctccaagtag cgaagcgagc aggactgggc
     6001 ggcggccaaa gcggtcggac agtgctccga gaacgggtgc gcatagaaat tgcatcaacg
     6061 catatagcgc tagcagcacg ccatagtgac tggcgatgct gtcggaatgg acgatatccc
     6121 gcaagaggcc cggcagtacc ggcataacca agcctatgcc tacagcatcc agggtgacgg
     6181 tgccgaggat gacgatgagc gcattgttag atttcataca cggtgcctga ctgcgttagc
     6241 aatttaactg tgataaacta ccgcattaaa gcttatcgat gataagctgt caaacatgag
     6301 aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     6361 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     6421 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     6481 atgcttcaat aacattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     6541 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     6601 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     6661 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     6721 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
     6781 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     6841 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     6901 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     6961 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     7021 ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa
     7081 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     7141 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     7201 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     7261 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     7321 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     7381 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     7441 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     7501 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     7561 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     7621 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     7681 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     7741 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     7801 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     7861 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     7921 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     7981 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     8041 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     8101 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     8161 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     8221 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     8281 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     8341 catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg
     8401 catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg
     8461 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     8521 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     8581 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
     8641 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
     8701 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt
     8761 gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac
     8821 gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact
     8881 ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt
     8941 taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa
     9001 cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa
     9061 gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg
     9121 ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt
     9181 cctcaacgac aggagcacga tcatgcgcac ccgtggccag gacccaacgc tgcccgagat
     9241 gcgccgcgtg cggctgctgg agatggcgga cgcgatggat atgttctgcc aagggttggt
     9301 ttgcgcattc acagttctcc gcaagaattg attggctcca attcttggag tggtgaatcc
     9361 gttagcgagg tgccgccggc ttccattcag gtcgaggtgg cccggctcca tgcaccgcga
     9421 cgcaacgcgg ggaggcagac aaggtatagg gcggcgccta caatccatgc caacccgttc
     9481 catgtgctcg ccgaggcggc ataaatcgcc gtgacgatca gcggtccaat gatcgaagtt
     9541 aggctggtaa gagccgcgag cgatccttga agctgtccct gatggtcgtc atctacctgc
     9601 ctggacagca tggcctgcaa cgcgggcatc ccgatgccgc cggaagcgag aagaatcata
     9661 atggggaagg ccatccagcc tcgcgtcgcg aacgccagca agacgtagcc cagcgcgtcg
     9721 gccgccatgc cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg
     9781 acgaaggctt gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc
     9841 gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt
     9901 cctacgagtt gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc
     9961 gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg acgctctccc
    10021 ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt gag
//
