pJSC119 - Bacterial expression plasmid for SpCas9 + N497A/R6

LOCUS       pJSC119 - Bacterial expression plasmid for SpCas9 + N497A/R661A/Q695A variant               10073 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Bacterial expression plasmid for SpCas9 + N497A/R661A/Q695A variant.
ACCESSION   .
VERSION     .
KEYWORDS    pJSC119 - Bacterial expression plasmid for SpCas9 +
            N497A/R661...
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 May 13, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..10073
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     86..105
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     189..208
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        189..207
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             296..318
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             338..367
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             377..1477
                     /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     1451..1474
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1532..1552
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1559..5665
                     /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
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTAFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGALSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     ALIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     primer_bind     complement(5791..5809)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5805..5852
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(6218..6246)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(6273..6291)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6359..6463
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6464..7324
                     /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(6682..6701)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7495..8083
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7984..8003
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8237..8254
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    8269..8408
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(8334..8353)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8494..8516
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(8510..8701)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
       61 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
      121 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atctcgatcc
      181 cgcgaaatta atacgactca ctatagggag accacaacgg tttccctcta gtgccggctc
      241 cggagagctc tttaattaag cggccgccct gcaggactcg agttctagaa ataattttgt
      301 ttaactttaa gaaggagata tacatatgaa atcttctcac catcaccatc accatcacca
      361 tcaccatggt tcttctatga aaatcgaaga aggtaaactg gtaatctgga ttaacggcga
      421 taaaggctat aacggtctcg ctgaagtcgg taagaaattc gagaaagata ccggaattaa
      481 agtcaccgtt gagcatccgg ataaactgga agagaaattc ccacaggttg cggcaactgg
      541 cgatggccct gacattatct tctgggcaca cgaccgcttt ggtggctacg ctcaatctgg
      601 cctgttggct gaaatcaccc cggacaaagc gttccaggac aagctgtatc cgtttacctg
      661 ggatgccgta cgttacaacg gcaagctgat tgcttacccg atcgctgttg aagcgttatc
      721 gctgatttat aacaaagatc tgctgccgaa cccgccaaaa acctgggaag agatcccggc
      781 gctggataaa gaactgaaag cgaaaggtaa gagcgcgctg atgttcaacc tgcaagaacc
      841 gtacttcacc tggccgctga ttgctgctga cgggggttat gcgttcaagt atgaaaacgg
      901 caagtacgac attaaagacg tgggcgtgga taacgctggc gcgaaagcgg gtctgacctt
      961 cctggttgac ctgattaaaa acaaacacat gaatgcagac accgattact ccatcgcaga
     1021 agctgccttt aataaaggcg aaacagcgat gaccatcaac ggcccgtggg catggtccaa
     1081 catcgacacc agcaaagtga attatggtgt aacggtactg ccgaccttca agggtcaacc
     1141 atccaaaccg ttcgttggcg tgctgagcgc aggtattaac gccgccagtc cgaacaaaga
     1201 gctggcaaaa gagttcctcg aaaactatct gctgactgat gaaggtctgg aagcggttaa
     1261 taaagacaaa ccgctgggtg ccgtagcgct gaagtcttac gaggaagagt tggcgaaaga
     1321 tccacgtatt gccgccacta tggaaaacgc ccagaaaggt gaaatcatgc cgaacatccc
     1381 gcagatgtcc gctttctggt atgccgtgcg tactgcggtg atcaacgccg ccagcggtcg
     1441 tcagactgtc gatgaagccc tgaaagacgc gcagactaat tcgagctcga acaacaacaa
     1501 caataacaat aacaacaacc tcgggatcga ggaaaacctg tacttccaat ccaatgcaat
     1561 ggataagaaa tactcaatag gcttagatat cggcacaaat agcgtcggat gggcggtgat
     1621 cactgatgaa tataaggttc cgtctaaaaa gttcaaggtt ctgggaaata cagaccgcca
     1681 cagtatcaaa aaaaatctta taggggctct tttatttgac agtggagaga cagcggaagc
     1741 gactcgtctc aaacggacag ctcgtagaag gtatacacgt cggaagaatc gtatttgtta
     1801 tctacaggag attttttcaa atgagatggc gaaagtagat gatagtttct ttcatcgact
     1861 tgaagagtct tttttggtgg aagaagacaa gaagcatgaa cgtcatccta tttttggaaa
     1921 tatagtagat gaagttgctt atcatgagaa atatccaact atctatcatc tgcgaaaaaa
     1981 attggtagat tctactgata aagcggattt gcgcttaatc tatttggcct tagcgcatat
     2041 gattaagttt cgtggtcatt ttttgattga gggagattta aatcctgata atagtgatgt
     2101 ggacaaacta tttatccagt tggtacaaac ctacaatcaa ttatttgaag aaaaccctat
     2161 taacgcaagt ggagtagatg ctaaagcgat tctttctgca cgattgagta aatcaagacg
     2221 attagaaaat ctcattgctc agctccccgg tgagaagaaa aatggcttat ttgggaatct
     2281 cattgctttg tcattgggtt tgacccctaa ttttaaatca aattttgatt tggcagaaga
     2341 tgctaaatta cagctttcaa aagatactta cgatgatgat ttagataatt tattggcgca
     2401 aattggagat caatatgctg atttgttttt ggcagctaag aatttatcag atgctatttt
     2461 actttcagat atcctaagag taaatactga aataactaag gctcccctat cagcttcaat
     2521 gattaaacgc tacgatgaac atcatcaaga cttgactctt ttaaaagctt tagttcgaca
     2581 acaacttcca gaaaagtata aagaaatctt ttttgatcaa tcaaaaaacg gatatgcagg
     2641 ttatattgat gggggagcta gccaagaaga attttataaa tttatcaaac caattttaga
     2701 aaaaatggat ggtactgagg aattattggt gaaactaaat cgtgaagatt tgctgcgcaa
     2761 gcaacggacc tttgacaacg gctctattcc ccatcaaatt cacttgggtg agctgcatgc
     2821 tattttgaga agacaagaag acttttatcc atttttaaaa gacaatcgtg agaagattga
     2881 aaaaatcttg acttttcgaa ttccttatta tgttggtcca ttggcgcgtg gcaatagtcg
     2941 ttttgcatgg atgactcgga agtctgaaga aacaattacc ccatggaatt ttgaagaagt
     3001 tgtcgataaa ggtgcttcag ctcaatcatt tattgaacgc atgacagcgt ttgataaaaa
     3061 tcttccaaat gaaaaagtac taccaaaaca tagtttgctt tatgagtatt ttacggttta
     3121 taacgaattg acaaaggtca aatatgttac tgaaggaatg cgaaaaccag catttctttc
     3181 aggtgaacag aagaaagcca ttgttgattt actcttcaaa acaaatcgaa aagtaaccgt
     3241 taagcaatta aaagaagatt atttcaaaaa aatagaatgt tttgatagtg ttgaaatttc
     3301 aggagttgaa gatagattta atgcttcatt aggtacctac catgatttgc taaaaattat
     3361 taaagataaa gattttttgg ataatgaaga aaatgaagat atcttagagg atattgtttt
     3421 aacattgacc ttatttgaag atagggagat gattgaggaa agacttaaaa catatgctca
     3481 cctctttgat gataaggtga tgaaacagct taaacgtcgc cgttatactg gttggggagc
     3541 gttgtctcga aaattgatta atggtattag ggataagcaa tctggcaaaa caatattaga
     3601 ttttttgaaa tcagatggtt ttgccaatcg caattttatg gccctgatcc atgatgatag
     3661 tttgacattt aaagaagaca ttcaaaaagc acaagtgtct ggacaaggcg atagtttaca
     3721 tgaacatatt gcaaatttag ctggtagccc tgctattaaa aaaggtattt tacagactgt
     3781 aaaagttgtt gatgaattgg tcaaagtaat ggggcggcat aagccagaaa atatcgttat
     3841 tgaaatggca cgtgaaaatc agacaactca aaagggccag aaaaattcgc gagagcgtat
     3901 gaaacgaatc gaagaaggta tcaaagaatt aggaagtcag attcttaaag agcatcctgt
     3961 tgaaaatact caattgcaaa atgaaaagct ctatctctat tatctccaaa atggaagaga
     4021 catgtatgtg gaccaagaat tagatattaa tcgtttaagt gattatgatg tcgatcacat
     4081 tgttccacaa agtttcctta aagacgattc aatagacaat aaggtcttaa cgcgttctga
     4141 taaaaatcgt ggtaaatcgg ataacgttcc aagtgaagaa gtagtcaaaa agatgaaaaa
     4201 ctattggaga caacttctaa acgccaagtt aatcactcaa cgtaagtttg ataatttaac
     4261 gaaagctgaa cgtggaggtt tgagtgaact tgataaagct ggttttatca aacgccaatt
     4321 ggttgaaact cgccaaatca ctaagcatgt ggcacaaatt ttggatagtc gcatgaatac
     4381 taaatacgat gaaaatgata aacttattcg agaggttaaa gtgattacct taaaatctaa
     4441 attagtttct gacttccgaa aagatttcca attctataaa gtacgtgaga ttaacaatta
     4501 ccatcatgcc catgatgcgt atctaaatgc cgtcgttgga actgctttga ttaagaaata
     4561 tccaaaactt gaatcggagt ttgtctatgg tgattataaa gtttatgatg ttcgtaaaat
     4621 gattgctaag tctgagcaag aaataggcaa agcaaccgca aaatatttct tttactctaa
     4681 tatcatgaac ttcttcaaaa cagaaattac acttgcaaat ggagagattc gcaaacgccc
     4741 tctaatcgaa actaatgggg aaactggaga aattgtctgg gataaagggc gagattttgc
     4801 cacagtgcgc aaagtattgt ccatgcccca agtcaatatt gtcaagaaaa cagaagtaca
     4861 gacaggcgga ttctccaagg agtcaatttt accaaaaaga aattcggaca agcttattgc
     4921 tcgtaaaaaa gactgggatc caaaaaaata tggtggtttt gatagtccaa cggtagctta
     4981 ttcagtccta gtggttgcta aggtggaaaa agggaaatcg aagaagttaa aatccgttaa
     5041 agagttacta gggatcacaa ttatggaaag aagttccttt gaaaaaaatc cgattgactt
     5101 tttagaagct aaaggatata aggaagttaa aaaagactta atcattaaac tacctaaata
     5161 tagtcttttt gagttagaaa acggtcgtaa acggatgctg gctagtgccg gagaattaca
     5221 aaaaggaaat gagctggctc tgccaagcaa atatgtgaat tttttatatt tagctagtca
     5281 ttatgaaaag ttgaagggta gtccagaaga taacgaacaa aaacaattgt ttgtggagca
     5341 gcataagcat tatttagatg agattattga gcaaatcagt gaattttcta agcgtgttat
     5401 tttagcagat gccaatttag ataaagttct tagtgcatat aacaaacata gagacaaacc
     5461 aatacgtgaa caagcagaaa atattattca tttatttacg ttgacgaatc ttggagctcc
     5521 cgctgctttt aaatattttg atacaacaat tgatcgtaaa cgatatacgt ctacaaaaga
     5581 agttttagat gccactctta tccatcaatc catcactggt ctttatgaaa cacgcattga
     5641 tttgagtcag ctaggaggtg actaataaca ttggaagtgg ataacggatc cgcgatcgcg
     5701 gcgcgccacc tggtggccgg ccggtaccac gcgtgcgcgc tgatccggct gctaacaaag
     5761 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
     5821 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggatatc
     5881 cacaggacgg gtgtggtcgc catgatcgcg tagtcgatag tggctccaag tagcgaagcg
     5941 agcaggactg ggcggcggcc aaagcggtcg gacagtgctc cgagaacggg tgcgcataga
     6001 aattgcatca acgcatatag cgctagcagc acgccatagt gactggcgat gctgtcggaa
     6061 tggacgatat cccgcaagag gcccggcagt accggcataa ccaagcctat gcctacagca
     6121 tccagggtga cggtgccgag gatgacgatg agcgcattgt tagatttcat acacggtgcc
     6181 tgactgcgtt agcaatttaa ctgtgataaa ctaccgcatt aaagcttatc gatgataagc
     6241 tgtcaaacat gagaattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
     6301 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     6361 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     6421 ataaccctga taaatgcttc aataacattg aaaaaggaag agtatgagta ttcaacattt
     6481 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     6541 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     6601 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     6661 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
     6721 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     6781 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     6841 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     6901 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     6961 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
     7021 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     7081 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     7141 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     7201 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     7261 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     7321 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     7381 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     7441 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     7501 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     7561 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     7621 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     7681 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     7741 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     7801 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     7861 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     7921 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     7981 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     8041 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     8101 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     8161 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     8221 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta
     8281 ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc
     8341 tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca
     8401 tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     8461 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     8521 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa
     8581 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg
     8641 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca
     8701 ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag
     8761 agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg
     8821 agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat
     8881 gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga
     8941 tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa
     9001 cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt
     9061 cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc
     9121 agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggc caggacccaa
     9181 cgctgcccga gatgcgccgc gtgcggctgc tggagatggc ggacgcgatg gatatgttct
     9241 gccaagggtt ggtttgcgca ttcacagttc tccgcaagaa ttgattggct ccaattcttg
     9301 gagtggtgaa tccgttagcg aggtgccgcc ggcttccatt caggtcgagg tggcccggct
     9361 ccatgcaccg cgacgcaacg cggggaggca gacaaggtat agggcggcgc ctacaatcca
     9421 tgccaacccg ttccatgtgc tcgccgaggc ggcataaatc gccgtgacga tcagcggtcc
     9481 aatgatcgaa gttaggctgg taagagccgc gagcgatcct tgaagctgtc cctgatggtc
     9541 gtcatctacc tgcctggaca gcatggcctg caacgcgggc atcccgatgc cgccggaagc
     9601 gagaagaatc ataatgggga aggccatcca gcctcgcgtc gcgaacgcca gcaagacgta
     9661 gcccagcgcg tcggccgcca tgccggcgat aatggcctgc ttctcgccga aacgtttggt
     9721 ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata ccgcaagcga
     9781 caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc
     9841 tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat
     9901 agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg
     9961 tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc
    10021 gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag atggcgccca aca
//