pJSC077 - Bacterial expression plasmid for SpCas9 + K855A, H

LOCUS       pJSC077 - Bacterial expression plasmid for SpCas9 + K855A, HNH FRET variant               10073 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Bacterial expression plasmid for SpCas9 + K855A, HNH FRET variant.
ACCESSION   .
VERSION     .
KEYWORDS    pJSC077 - Bacterial expression plasmid for SpCas9 + K855A, HNH...
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     543..562
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     646..665
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        646..664
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             753..775
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             795..824
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             834..1934
                     /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     1908..1931
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1989..2009
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2016..6122
                     /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
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIESFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNAVLTRSDKNRGKCDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     primer_bind     complement(6248..6266)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6262..6309
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(6675..6703)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(6730..6748)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6816..6920
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6921..7781
                     /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(7139..7158)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7952..8540
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8441..8460
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8694..8711
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    8726..8865
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(8791..8810)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8951..8973
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(8967..9158)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gggaaggcca tccagcctcg cgtcgcgaac gccagcaaga cgtagcccag cgcgtcggcc
       61 gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg
      121 aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc
      181 gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct
      241 acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc
      301 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgacg ctctccctta
      361 tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc
      421 gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc
      481 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca
      541 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc
      601 acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc gaaattaata cgactcacta
      661 tagggagacc acaacggttt ccctctagtg ccggctccgg agagctcttt aattaagcgg
      721 ccgccctgca ggactcgagt tctagaaata attttgttta actttaagaa ggagatatac
      781 atatgaaatc ttctcaccat caccatcacc atcaccatca ccatggttct tctatgaaaa
      841 tcgaagaagg taaactggta atctggatta acggcgataa aggctataac ggtctcgctg
      901 aagtcggtaa gaaattcgag aaagataccg gaattaaagt caccgttgag catccggata
      961 aactggaaga gaaattccca caggttgcgg caactggcga tggccctgac attatcttct
     1021 gggcacacga ccgctttggt ggctacgctc aatctggcct gttggctgaa atcaccccgg
     1081 acaaagcgtt ccaggacaag ctgtatccgt ttacctggga tgccgtacgt tacaacggca
     1141 agctgattgc ttacccgatc gctgttgaag cgttatcgct gatttataac aaagatctgc
     1201 tgccgaaccc gccaaaaacc tgggaagaga tcccggcgct ggataaagaa ctgaaagcga
     1261 aaggtaagag cgcgctgatg ttcaacctgc aagaaccgta cttcacctgg ccgctgattg
     1321 ctgctgacgg gggttatgcg ttcaagtatg aaaacggcaa gtacgacatt aaagacgtgg
     1381 gcgtggataa cgctggcgca aaagcgggtc tgaccttcct ggttgacctg attaaaaaca
     1441 aacacatgaa tgcagacacc gattactcca tcgcagaagc tgcctttaat aaaggcgaaa
     1501 cagcgatgac catcaacggc ccgtgggcat ggtccaacat cgacaccagc aaagtgaatt
     1561 atggtgtaac ggtactgccg accttcaagg gtcaaccatc caaaccgttc gttggcgtgc
     1621 tgagcgcagg tattaacgcc gccagtccga acaaagagct ggcaaaagag ttcctcgaaa
     1681 actatctgct gactgatgaa ggtctggaag cggttaataa agacaaaccg ctgggtgccg
     1741 tagcgctgaa gtcttacgag gaagagttgg cgaaagatcc acgtattgcc gccactatgg
     1801 aaaacgccca gaaaggtgaa atcatgccga acatcccgca gatgtccgct ttctggtatg
     1861 ccgtgcgtac tgcggtgatc aacgccgcca gcggtcgtca gactgtcgat gaagccctga
     1921 aagacgcgca gactaattcg agctcgaaca acaacaacaa taacaataac aacaacctcg
     1981 ggatcgagga aaacctgtac ttccaatcca atgcaatgga taagaaatac tcaataggct
     2041 tagatatcgg cacaaatagc gtcggatggg cggtgatcac tgatgaatat aaggttccgt
     2101 ctaaaaagtt caaggttctg ggaaatacag accgccacag tatcaaaaaa aatcttatag
     2161 gggctctttt atttgacagt ggagagacag cggaagcgac tcgtctcaaa cggacagctc
     2221 gtagaaggta tacacgtcgg aagaatcgta tttcttatct acaggagatt ttttcaaatg
     2281 agatggcgaa agtagatgat agtttctttc atcgacttga agagtctttt ttggtggaag
     2341 aagacaagaa gcatgaacgt catcctattt ttggaaatat agtagatgaa gttgcttatc
     2401 atgagaaata tccaactatc tatcatctgc gaaaaaaatt ggtagattct actgataaag
     2461 cggatttgcg cttaatctat ttggccttag cgcatatgat taagtttcgt ggtcattttt
     2521 tgattgaggg agatttaaat cctgataata gtgatgtgga caaactattt atccagttgg
     2581 tacaaaccta caatcaatta tttgaagaaa accctattaa cgcaagtgga gtagatgcta
     2641 aagcgattct ttctgcacga ttgagtaaat caagacgatt agaaaatctc attgctcagc
     2701 tccccggtga gaagaaaaat ggcttatttg ggaatctcat tgctttgtca ttgggtttga
     2761 cccctaattt taaatcaaat tttgatttgg cagaagatgc taaattacag ctttcaaaag
     2821 atacttacga tgatgattta gataatttat tggcgcaaat tggagatcaa tatgctgatt
     2881 tgtttttggc agctaagaat ttatcagatg ctattttact ttcagatatc ctaagagtaa
     2941 atactgaaat aactaaggct cccctatcag cttcaatgat taaacgctac gatgaacatc
     3001 atcaagactt gactctttta aaagctttag ttcgacaaca acttccagaa aagtataaag
     3061 aaatcttttt tgatcaatgt aaaaacggat atgcaggtta tattgatggg ggagctagcc
     3121 aagaagaatt ttataaattt atcaaaccaa ttttagaaaa aatggatggt actgaggaat
     3181 tattggtgaa actaaatcgt gaagatttgc tgcgcaagca acggaccttt gacaacggct
     3241 ctattcccca tcaaattcac ttgggtgagc tgcatgctat tttgagaaga caagaagact
     3301 tttatccatt tttaaaagac aatcgtgaga agattgaaaa aatcttgact tttcgaattc
     3361 cttattatgt tggtccattg gcgcgtggca atagtcgttt tgcatggatg actcggaagt
     3421 ctgaagaaac aattacccca tggaattttg aagaagttgt cgataaaggt gcttcagctc
     3481 aatcatttat tgaacgcatg acaaactttg ataaaaatct tccaaatgaa aaagtactac
     3541 caaaacatag tttgctttat gagtatttta cggtttataa cgaattgaca aaggtcaaat
     3601 atgttactga aggaatgcga aaaccagcat ttctttcagg tgaacagaag aaagccattg
     3661 ttgatttact cttcaaaaca aatcgaaaag taaccgttaa gcaattaaaa gaagattatt
     3721 tcaaaaaaat agaatctttt gatagtgttg aaatttcagg agttgaagat agatttaatg
     3781 cttcattagg tacctaccat gatttgctaa aaattattaa agataaagat tttttggata
     3841 atgaagaaaa tgaagatatc ttagaggata ttgttttaac attgacctta tttgaagata
     3901 gggagatgat tgaggaaaga cttaaaacat atgctcacct ctttgatgat aaggtgatga
     3961 aacagcttaa acgtcgccgt tatactggtt ggggacgttt gtctcgaaaa ttgattaatg
     4021 gtattaggga taagcaatct ggcaaaacaa tattagattt tttgaaatca gatggttttg
     4081 ccaatcgcaa ttttatgcag ctgatccatg atgatagttt gacatttaaa gaagacattc
     4141 aaaaagcaca agtgtctgga caaggcgata gtttacatga acatattgca aatttagctg
     4201 gtagccctgc tattaaaaaa ggtattttac agactgtaaa agttgttgat gaattggtca
     4261 aagtaatggg gcggcataag ccagaaaata tcgttattga aatggcacgt gaaaatcaga
     4321 caactcaaaa gggccagaaa aattcgcgag agcgtatgaa acgaatcgaa gaaggtatca
     4381 aagaattagg aagtcagatt cttaaagagc atcctgttga aaatactcaa ttgcaaaatg
     4441 aaaagctcta tctctattat ctccaaaatg gaagagacat gtatgtggac caagaattag
     4501 atattaatcg tttaagtgat tatgatgtcg atcacattgt tccacaaagt ttccttaaag
     4561 acgattcaat agacaatgct gtcttaacgc gttctgataa aaatcgtggt aaatgcgata
     4621 acgttccaag tgaagaagta gtcaaaaaga tgaaaaacta ttggagacaa cttctaaacg
     4681 ccaagttaat cactcaacgt aagtttgata atttaacgaa agctgaacgt ggaggtttga
     4741 gtgaacttga taaagctggt tttatcaaac gccaattggt tgaaactcgc caaatcacta
     4801 agcatgtggc acaaattttg gatagtcgca tgaatactaa atacgatgaa aatgataaac
     4861 ttattcgaga ggttaaagtg attaccttaa aatctaaatt agtttctgac ttccgaaaag
     4921 atttccaatt ctataaagta cgtgagatta acaattacca tcatgcccat gatgcgtatc
     4981 taaatgccgt cgttggaact gctttgatta agaaatatcc aaaacttgaa tcggagtttg
     5041 tctatggtga ttataaagtt tatgatgttc gtaaaatgat tgctaagtct gagcaagaaa
     5101 taggcaaagc aaccgcaaaa tatttctttt actctaatat catgaacttc ttcaaaacag
     5161 aaattacact tgcaaatgga gagattcgca aacgccctct aatcgaaact aatggggaaa
     5221 ctggagaaat tgtctgggat aaagggcgag attttgccac agtgcgcaaa gtattgtcca
     5281 tgccccaagt caatattgtc aagaaaacag aagtacagac aggcggattc tccaaggagt
     5341 caattttacc aaaaagaaat tcggacaagc ttattgctcg taaaaaagac tgggatccaa
     5401 aaaaatatgg tggttttgat agtccaacgg tagcttattc agtcctagtg gttgctaagg
     5461 tggaaaaagg gaaatcgaag aagttaaaat ccgttaaaga gttactaggg atcacaatta
     5521 tggaaagaag ttcctttgaa aaaaatccga ttgacttttt agaagctaaa ggatataagg
     5581 aagttaaaaa agacttaatc attaaactac ctaaatatag tctttttgag ttagaaaacg
     5641 gtcgtaaacg gatgctggct agtgccggag aattacaaaa aggaaatgag ctggctctgc
     5701 caagcaaata tgtgaatttt ttatatttag ctagtcatta tgaaaagttg aagggtagtc
     5761 cagaagataa cgaacaaaaa caattgtttg tggagcagca taagcattat ttagatgaga
     5821 ttattgagca aatcagtgaa ttttctaagc gtgttatttt agcagatgcc aatttagata
     5881 aagttcttag tgcatataac aaacatagag acaaaccaat acgtgaacaa gcagaaaata
     5941 ttattcattt atttacgttg acgaatcttg gagctcccgc tgcttttaaa tattttgata
     6001 caacaattga tcgtaaacga tatacgtcta caaaagaagt tttagatgcc actcttatcc
     6061 atcaatccat cactggtctt tatgaaacac gcattgattt gagtcagcta ggaggtgact
     6121 aataacattg gaagtggata acggatccgc gatcgcggcg cgccacctgg tggccggccg
     6181 gtaccacgcg tgcgctctga tccggctgct aacaaagccc gaaaggaagc tgagttggct
     6241 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
     6301 ggttttttgc tgaaaggagg aactatatcc ggatatccac aggacgggtg tggtcgccat
     6361 gatcgcgtag tcgatagtgg ctccaagtag cgaagcgagc aggactgggc ggcggccaaa
     6421 gcggtcggac agtgctccga gaacgggtgc gcatagaaat tgcatcaacg catatagcgc
     6481 tagcagcacg ccatagtgac tggcgatgct gtcggaatgg acgatatccc gcaagaggcc
     6541 cggcagtacc ggcataacca agcctatgcc tacagcatcc agggtgacgg tgccgaggat
     6601 gacgatgagc gcattgttag atttcataca cggtgcctga ctgcgttagc aatttaactg
     6661 tgataaacta ccgcattaaa gcttatcgat gataagctgt caaacatgag aattcttgaa
     6721 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     6781 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     6841 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     6901 aacattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     6961 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     7021 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     7081 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     7141 tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac
     7201 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     7261 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     7321 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     7381 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     7441 acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg
     7501 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     7561 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     7621 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     7681 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     7741 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     7801 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     7861 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     7921 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     7981 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     8041 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     8101 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     8161 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     8221 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     8281 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     8341 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     8401 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     8461 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     8521 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     8581 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     8641 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     8701 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     8761 gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     8821 ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca
     8881 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     8941 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     9001 aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt
     9061 tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag
     9121 cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga
     9181 tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt
     9241 actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg
     9301 atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat
     9361 gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg
     9421 cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat
     9481 gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc
     9541 ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac
     9601 aggagcacga tcatgcgcac ccgtggccag gacccaacgc tgcccgagat gcgccgcgtg
     9661 cggctgctgg agatggcgga cgcgatggat atgttctgcc aagggttggt ttgcgcattc
     9721 acagttctcc gcaagaattg attggctcca attcttggag tggtgaatcc gttagcgagg
     9781 tgccgccggc ttccattcag gtcgaggtgg cccggctcca tgcaccgcga cgcaacgcgg
     9841 ggaggcagac aaggtatagg gcggcgccta caatccatgc caacccgttc catgtgctcg
     9901 ccgaggcggc ataaatcgcc gtgacgatca gcggtccaat gatcgaagtt aggctggtaa
     9961 gagccgcgag cgatccttga agctgtccct gatggtcgtc atctacctgc ctggacagca
    10021 tggcctgcaa cgcgggcatc ccgatgccgc cggaagcgag aagaatcata atg
//