LOCUS       Exported               10073 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Bacterial expression plasmid for SpCas9 + K855A variant.
ACCESSION   .
VERSION     .
KEYWORDS    pJSC090 - Bacterial expression plasmid for SpCas9 + K855A variant
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.snapgene.com
FEATURES             Location/Qualifiers
     source          1..10073
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     805..824
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     908..927
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        908..926
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1015..1037
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1057..1086
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             1096..2196
                     /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     2170..2193
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             2251..2271
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2278..6384
                     /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
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNAVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     primer_bind     complement(6510..6528)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6524..6571
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(6937..6965)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(6992..7010)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7078..7182
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7183..8043
                     /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(7401..7420)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8214..8802
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8703..8722
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8956..8973
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    8988..9127
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(9053..9072)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9213..9235
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(9229..9420)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 ccggctccat gcaccgcgac gcaacgcggg gaggcagaca aggtataggg cggcgcctac
       61 aatccatgcc aacccgttcc atgtgctcgc cgaggcggca taaatcgccg tgacgatcag
      121 cggtccaatg atcgaagtta ggctggtaag agccgcgagc gatccttgaa gctgtccctg
      181 atggtcgtca tctacctgcc tggacagcat ggcctgcaac gcgggcatcc cgatgccgcc
      241 ggaagcgaga agaatcataa tggggaaggc catccagcct cgcgtcgcga acgccagcaa
      301 gacgtagccc agcgcgtcgg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg
      361 tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc
      421 aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca
      481 gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc
      541 gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
      601 catcggtcga cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt
      661 gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag
      721 tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa
      781 gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc
      841 tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tctcgatccc
      901 gcgaaattaa tacgactcac tatagggaga ccacaacggt ttccctctag tgccggctcc
      961 ggagagctct ttaattaagc ggccgccctg caggactcga gttctagaaa taattttgtt
     1021 taactttaag aaggagatat acatatgaaa tcttctcacc atcaccatca ccatcaccat
     1081 caccatggtt cttctatgaa aatcgaagaa ggtaaactgg taatctggat taacggcgat
     1141 aaaggctata acggtctcgc tgaagtcggt aagaaattcg agaaagatac cggaattaaa
     1201 gtcaccgttg agcatccgga taaactggaa gagaaattcc cacaggttgc ggcaactggc
     1261 gatggccctg acattatctt ctgggcacac gaccgctttg gtggctacgc tcaatctggc
     1321 ctgttggctg aaatcacccc ggacaaagcg ttccaggaca agctgtatcc gtttacctgg
     1381 gatgccgtac gttacaacgg caagctgatt gcttacccga tcgctgttga agcgttatcg
     1441 ctgatttata acaaagatct gctgccgaac ccgccaaaaa cctgggaaga gatcccggcg
     1501 ctggataaag aactgaaagc gaaaggtaag agcgcgctga tgttcaacct gcaagaaccg
     1561 tacttcacct ggccgctgat tgctgctgac gggggttatg cgttcaagta tgaaaacggc
     1621 aagtacgaca ttaaagacgt gggcgtggat aacgctggcg cgaaagcggg tctgaccttc
     1681 ctggttgacc tgattaaaaa caaacacatg aatgcagaca ccgattactc catcgcagaa
     1741 gctgccttta ataaaggcga aacagcgatg accatcaacg gcccgtgggc atggtccaac
     1801 atcgacacca gcaaagtgaa ttatggtgta acggtactgc cgaccttcaa gggtcaacca
     1861 tccaaaccgt tcgttggcgt gctgagcgca ggtattaacg ccgccagtcc gaacaaagag
     1921 ctggcaaaag agttcctcga aaactatctg ctgactgatg aaggtctgga agcggttaat
     1981 aaagacaaac cgctgggtgc cgtagcgctg aagtcttacg aggaagagtt ggcgaaagat
     2041 ccacgtattg ccgccactat ggaaaacgcc cagaaaggtg aaatcatgcc gaacatcccg
     2101 cagatgtccg ctttctggta tgccgtgcgt actgcggtga tcaacgccgc cagcggtcgt
     2161 cagactgtcg atgaagccct gaaagacgcg cagactaatt cgagctcgaa caacaacaac
     2221 aataacaata acaacaacct cgggatcgag gaaaacctgt acttccaatc caatgcaatg
     2281 gataagaaat actcaatagg cttagatatc ggcacaaata gcgtcggatg ggcggtgatc
     2341 actgatgaat ataaggttcc gtctaaaaag ttcaaggttc tgggaaatac agaccgccac
     2401 agtatcaaaa aaaatcttat aggggctctt ttatttgaca gtggagagac agcggaagcg
     2461 actcgtctca aacggacagc tcgtagaagg tatacacgtc ggaagaatcg tatttgttat
     2521 ctacaggaga ttttttcaaa tgagatggcg aaagtagatg atagtttctt tcatcgactt
     2581 gaagagtctt ttttggtgga agaagacaag aagcatgaac gtcatcctat ttttggaaat
     2641 atagtagatg aagttgctta tcatgagaaa tatccaacta tctatcatct gcgaaaaaaa
     2701 ttggtagatt ctactgataa agcggatttg cgcttaatct atttggcctt agcgcatatg
     2761 attaagtttc gtggtcattt tttgattgag ggagatttaa atcctgataa tagtgatgtg
     2821 gacaaactat ttatccagtt ggtacaaacc tacaatcaat tatttgaaga aaaccctatt
     2881 aacgcaagtg gagtagatgc taaagcgatt ctttctgcac gattgagtaa atcaagacga
     2941 ttagaaaatc tcattgctca gctccccggt gagaagaaaa atggcttatt tgggaatctc
     3001 attgctttgt cattgggttt gacccctaat tttaaatcaa attttgattt ggcagaagat
     3061 gctaaattac agctttcaaa agatacttac gatgatgatt tagataattt attggcgcaa
     3121 attggagatc aatatgctga tttgtttttg gcagctaaga atttatcaga tgctatttta
     3181 ctttcagata tcctaagagt aaatactgaa ataactaagg ctcccctatc agcttcaatg
     3241 attaaacgct acgatgaaca tcatcaagac ttgactcttt taaaagcttt agttcgacaa
     3301 caacttccag aaaagtataa agaaatcttt tttgatcaat caaaaaacgg atatgcaggt
     3361 tatattgatg ggggagctag ccaagaagaa ttttataaat ttatcaaacc aattttagaa
     3421 aaaatggatg gtactgagga attattggtg aaactaaatc gtgaagattt gctgcgcaag
     3481 caacggacct ttgacaacgg ctctattccc catcaaattc acttgggtga gctgcatgct
     3541 attttgagaa gacaagaaga cttttatcca tttttaaaag acaatcgtga gaagattgaa
     3601 aaaatcttga cttttcgaat tccttattat gttggtccat tggcgcgtgg caatagtcgt
     3661 tttgcatgga tgactcggaa gtctgaagaa acaattaccc catggaattt tgaagaagtt
     3721 gtcgataaag gtgcttcagc tcaatcattt attgaacgca tgacaaactt tgataaaaat
     3781 cttccaaatg aaaaagtact accaaaacat agtttgcttt atgagtattt tacggtttat
     3841 aacgaattga caaaggtcaa atatgttact gaaggaatgc gaaaaccagc atttctttca
     3901 ggtgaacaga agaaagccat tgttgattta ctcttcaaaa caaatcgaaa agtaaccgtt
     3961 aagcaattaa aagaagatta tttcaaaaaa atagaatgtt ttgatagtgt tgaaatttca
     4021 ggagttgaag atagatttaa tgcttcatta ggtacctacc atgatttgct aaaaattatt
     4081 aaagataaag attttttgga taatgaagaa aatgaagata tcttagagga tattgtttta
     4141 acattgacct tatttgaaga tagggagatg attgaggaaa gacttaaaac atatgctcac
     4201 ctctttgatg ataaggtgat gaaacagctt aaacgtcgcc gttatactgg ttggggacgt
     4261 ttgtctcgaa aattgattaa tggtattagg gataagcaat ctggcaaaac aatattagat
     4321 tttttgaaat cagatggttt tgccaatcgc aattttatgc agctgatcca tgatgatagt
     4381 ttgacattta aagaagacat tcaaaaagca caagtgtctg gacaaggcga tagtttacat
     4441 gaacatattg caaatttagc tggtagccct gctattaaaa aaggtatttt acagactgta
     4501 aaagttgttg atgaattggt caaagtaatg gggcggcata agccagaaaa tatcgttatt
     4561 gaaatggcac gtgaaaatca gacaactcaa aagggccaga aaaattcgcg agagcgtatg
     4621 aaacgaatcg aagaaggtat caaagaatta ggaagtcaga ttcttaaaga gcatcctgtt
     4681 gaaaatactc aattgcaaaa tgaaaagctc tatctctatt atctccaaaa tggaagagac
     4741 atgtatgtgg accaagaatt agatattaat cgtttaagtg attatgatgt cgatcacatt
     4801 gttccacaaa gtttccttaa agacgattca atagacaatg ctgtcttaac gcgttctgat
     4861 aaaaatcgtg gtaaatcgga taacgttcca agtgaagaag tagtcaaaaa gatgaaaaac
     4921 tattggagac aacttctaaa cgccaagtta atcactcaac gtaagtttga taatttaacg
     4981 aaagctgaac gtggaggttt gagtgaactt gataaagctg gttttatcaa acgccaattg
     5041 gttgaaactc gccaaatcac taagcatgtg gcacaaattt tggatagtcg catgaatact
     5101 aaatacgatg aaaatgataa acttattcga gaggttaaag tgattacctt aaaatctaaa
     5161 ttagtttctg acttccgaaa agatttccaa ttctataaag tacgtgagat taacaattac
     5221 catcatgccc atgatgcgta tctaaatgcc gtcgttggaa ctgctttgat taagaaatat
     5281 ccaaaacttg aatcggagtt tgtctatggt gattataaag tttatgatgt tcgtaaaatg
     5341 attgctaagt ctgagcaaga aataggcaaa gcaaccgcaa aatatttctt ttactctaat
     5401 atcatgaact tcttcaaaac agaaattaca cttgcaaatg gagagattcg caaacgccct
     5461 ctaatcgaaa ctaatgggga aactggagaa attgtctggg ataaagggcg agattttgcc
     5521 acagtgcgca aagtattgtc catgccccaa gtcaatattg tcaagaaaac agaagtacag
     5581 acaggcggat tctccaagga gtcaatttta ccaaaaagaa attcggacaa gcttattgct
     5641 cgtaaaaaag actgggatcc aaaaaaatat ggtggttttg atagtccaac ggtagcttat
     5701 tcagtcctag tggttgctaa ggtggaaaaa gggaaatcga agaagttaaa atccgttaaa
     5761 gagttactag ggatcacaat tatggaaaga agttcctttg aaaaaaatcc gattgacttt
     5821 ttagaagcta aaggatataa ggaagttaaa aaagacttaa tcattaaact acctaaatat
     5881 agtctttttg agttagaaaa cggtcgtaaa cggatgctgg ctagtgccgg agaattacaa
     5941 aaaggaaatg agctggctct gccaagcaaa tatgtgaatt ttttatattt agctagtcat
     6001 tatgaaaagt tgaagggtag tccagaagat aacgaacaaa aacaattgtt tgtggagcag
     6061 cataagcatt atttagatga gattattgag caaatcagtg aattttctaa gcgtgttatt
     6121 ttagcagatg ccaatttaga taaagttctt agtgcatata acaaacatag agacaaacca
     6181 atacgtgaac aagcagaaaa tattattcat ttatttacgt tgacgaatct tggagctccc
     6241 gctgctttta aatattttga tacaacaatt gatcgtaaac gatatacgtc tacaaaagaa
     6301 gttttagatg ccactcttat ccatcaatcc atcactggtc tttatgaaac acgcattgat
     6361 ttgagtcagc taggaggtga ctaataacat tggaagtgga taacggatcc gcgatcgcgg
     6421 cgcgccacct ggtggccggc cggtaccacg cgtgcgcgct gatccggctg ctaacaaagc
     6481 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
     6541 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggatatcc
     6601 acaggacggg tgtggtcgcc atgatcgcgt agtcgatagt ggctccaagt agcgaagcga
     6661 gcaggactgg gcggcggcca aagcggtcgg acagtgctcc gagaacgggt gcgcatagaa
     6721 attgcatcaa cgcatatagc gctagcagca cgccatagtg actggcgatg ctgtcggaat
     6781 ggacgatatc ccgcaagagg cccggcagta ccggcataac caagcctatg cctacagcat
     6841 ccagggtgac ggtgccgagg atgacgatga gcgcattgtt agatttcata cacggtgcct
     6901 gactgcgtta gcaatttaac tgtgataaac taccgcatta aagcttatcg atgataagct
     6961 gtcaaacatg agaattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt
     7021 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     7081 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     7141 taaccctgat aaatgcttca ataacattga aaaaggaaga gtatgagtat tcaacatttc
     7201 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     7261 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     7321 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     7381 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa
     7441 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     7501 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     7561 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     7621 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     7681 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca
     7741 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     7801 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     7861 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     7921 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     7981 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     8041 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     8101 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     8161 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     8221 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     8281 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     8341 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     8401 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     8461 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     8521 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     8581 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     8641 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     8701 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     8761 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     8821 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     8881 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     8941 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     9001 tttctcctta cgcatctgtg cggtatttca caccgcaatg gtgcactctc agtacaatct
     9061 gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat
     9121 ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     9181 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     9241 accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag
     9301 cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt
     9361 taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac
     9421 tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga
     9481 gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga
     9541 gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg
     9601 ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat
     9661 gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac tttacgaaac
     9721 acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc
     9781 gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca
     9841 gcctagccgg gtcctcaacg acaggagcac gatcatgcgc acccgtggcc aggacccaac
     9901 gctgcccgag atgcgccgcg tgcggctgct ggagatggcg gacgcgatgg atatgttctg
     9961 ccaagggttg gtttgcgcat tcacagttct ccgcaagaat tgattggctc caattcttgg
    10021 agtggtgaat ccgttagcga ggtgccgccg gcttccattc aggtcgaggt ggc
//
