LOCUS       sequence-177417-       10844 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of modified SpyCas9 in e.coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10844)
  AUTHORS   Staahl BT, Benekareddy M, Coulon-Bainier C, Banfal AA, Floor SN, 
            Sabo JK, Urnes C, Munares GA, Ghosh A, Doudna JA
  TITLE     Efficient genome editing in the mouse brain by local delivery of 
            engineered Cas9 ribonucleoprotein complexes.
  JOURNAL   Nat Biotechnol. 2017 Feb 13. doi: 10.1038/nbt.3806.
  PUBMED    28191903
REFERENCE   2  (bases 1 to 10844)
  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: "Nat 
            Biotechnol. 2017 Feb 13. doi: 10.1038/nbt.3806."
COMMENT     Sequence Label: pMJ915v2 + sfGFP
FEATURES             Location/Qualifiers
     source          1..10844
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1018..1041
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1099..1119
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1135..5238
                     /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
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             1135..5238
                     /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
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5245..5265
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5272..5292
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6141..6159)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6155..6202
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(6568..6573,6574..6590,6591..6596))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(6623..6641)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6709..6813
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6814..6882,6883..7674)
                     /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(7032..7051)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7845..8433
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8334..8353
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8587..8604
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    8619..8758
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(8684..8703)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8844..8866
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(8860..9051)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     10509..10528
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     10612..10631
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        10612..10630
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             10719..10741
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             10761..10778
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             join(10788..10844,1..1044)
                     /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"
ORIGIN
        1 ggtctcgctg aagtcggtaa gaaattcgag aaagataccg gaattaaagt caccgttgag
       61 catccggata aactggaaga gaaattccca caggttgcgg caactggcga tggccctgac
      121 attatcttct gggcacacga ccgctttggt ggctacgctc aatctggcct gttggctgaa
      181 atcaccccgg acaaagcgtt ccaggacaag ctgtatccgt ttacctggga tgccgtacgt
      241 tacaacggca agctgattgc ttacccgatc gctgttgaag cgttatcgct gatttataac
      301 aaagatctgc tgccgaaccc gccaaaaacc tgggaagaga tcccggcgct ggataaagaa
      361 ctgaaagcga aaggtaagag cgcgctgatg ttcaacctgc aagaaccgta cttcacctgg
      421 ccgctgattg ctgctgacgg gggttatgcg ttcaagtatg aaaacggcaa gtacgacatt
      481 aaagacgtgg gcgtggataa cgctggcgcg aaagcgggtc tgaccttcct ggttgacctg
      541 attaaaaaca aacacatgaa tgcagacacc gattactcca tcgcagaagc tgcctttaat
      601 aaaggcgaaa cagcgatgac catcaacggc ccgtgggcat ggtccaacat cgacaccagc
      661 aaagtgaatt atggtgtaac ggtactgccg accttcaagg gtcaaccatc caaaccgttc
      721 gttggcgtgc tgagcgcagg tattaacgcc gccagtccga acaaagagct ggcaaaagag
      781 ttcctcgaaa actatctgct gactgatgaa ggtctggaag cggttaataa agacaaaccg
      841 ctgggtgccg tagcgctgaa gtcttacgag gaagagttgg cgaaagatcc acgtattgcc
      901 gccactatgg aaaacgccca gaaaggtgaa atcatgccga acatcccgca gatgtccgct
      961 ttctggtatg ccgtgcgtac tgcggtgatc aacgccgcca gcggtcgtca gactgtcgat
     1021 gaagccctga aagacgcgca gactaattcg agctcgaaca acaacaacaa taacaataac
     1081 aacaacctcg ggatcgagga aaacctgtac ttccaatcca atgccactag taccatggat
     1141 aagaaatact caataggctt agatatcggc acaaatagcg tcggatgggc ggtgatcact
     1201 gatgaatata aggttccgtc taaaaagttc aaggttctgg gaaatacaga ccgccacagt
     1261 atcaaaaaaa atcttatagg ggctctttta tttgacagtg gagagacagc ggaagcgact
     1321 cgtctcaaac ggacagctcg tagaaggtat acacgtcgga agaatcgtat ttgttatcta
     1381 caggagattt tttcaaatga gatggcgaaa gtagatgata gtttctttca tcgacttgaa
     1441 gagtcttttt tggtggaaga agacaagaag catgaacgtc atcctatttt tggaaatata
     1501 gtagatgaag ttgcttatca tgagaaatat ccaactatct atcatctgcg aaaaaaattg
     1561 gtagattcta ctgataaagc ggatttgcgc ttaatctatt tggccttagc gcatatgatt
     1621 aagtttcgtg gtcatttttt gattgaggga gatttaaatc ctgataatag tgatgtggac
     1681 aaactattta tccagttggt acaaacctac aatcaattat ttgaagaaaa ccctattaac
     1741 gcaagtggag tagatgctaa agcgattctt tctgcacgat tgagtaaatc aagacgatta
     1801 gaaaatctca ttgctcagct ccccggtgag aagaaaaatg gcttatttgg gaatctcatt
     1861 gctttgtcat tgggtttgac ccctaatttt aaatcaaatt ttgatttggc agaagatgct
     1921 aaattacagc tttcaaaaga tacttacgat gatgatttag ataatttatt ggcgcaaatt
     1981 ggagatcaat atgctgattt gtttttggca gctaagaatt tatcagatgc tattttactt
     2041 tcagatatcc taagagtaaa tactgaaata actaaggctc ccctatcagc ttcaatgatt
     2101 aaacgctacg atgaacatca tcaagacttg actcttttaa aagctttagt tcgacaacaa
     2161 cttccagaaa agtataaaga aatctttttt gatcaatcaa aaaacggata tgcaggttat
     2221 attgatgggg gagctagcca agaagaattt tataaattta tcaaaccaat tttagaaaaa
     2281 atggatggta ctgaggaatt attggtgaaa ctaaatcgtg aagatttgct gcgcaagcaa
     2341 cggacctttg acaacggctc tattccccat caaattcact tgggtgagct gcatgctatt
     2401 ttgagaagac aagaagactt ttatccattt ttaaaagaca atcgtgagaa gattgaaaaa
     2461 atcttgactt ttcgaattcc ttattatgtt ggtccattgg cgcgtggcaa tagtcgtttt
     2521 gcatggatga ctcggaagtc tgaagaaaca attaccccat ggaattttga agaagttgtc
     2581 gataaaggtg cttcagctca atcatttatt gaacgcatga caaactttga taaaaatctt
     2641 ccaaatgaaa aagtactacc aaaacatagt ttgctttatg agtattttac ggtttataac
     2701 gaattgacaa aggtcaaata tgttactgaa ggaatgcgaa aaccagcatt tctttcaggt
     2761 gaacagaaga aagccattgt tgatttactc ttcaaaacaa atcgaaaagt aaccgttaag
     2821 caattaaaag aagattattt caaaaaaata gaatgttttg atagtgttga aatttcagga
     2881 gttgaagata gatttaatgc ttcattaggt acctaccatg atttgctaaa aattattaaa
     2941 gataaagatt ttttggataa tgaagaaaat gaagatatct tagaggatat tgttttaaca
     3001 ttgaccttat ttgaagatag ggagatgatt gaggaaagac ttaaaacata tgctcacctc
     3061 tttgatgata aggtgatgaa acagcttaaa cgtcgccgtt atactggttg gggacgtttg
     3121 tctcgaaaat tgattaatgg tattagggat aagcaatctg gcaaaacaat attagatttt
     3181 ttgaaatcag atggttttgc caatcgcaat tttatgcagc tgatccatga tgatagtttg
     3241 acatttaaag aagacattca aaaagcacaa gtgtctggac aaggcgatag tttacatgaa
     3301 catattgcaa atttagctgg tagccctgct attaaaaaag gtattttaca gactgtaaaa
     3361 gttgttgatg aattggtcaa agtaatgggg cggcataagc cagaaaatat cgttattgaa
     3421 atggcacgtg aaaatcagac aactcaaaag ggccagaaaa attcgcgaga gcgtatgaaa
     3481 cgaatcgaag aaggtatcaa agaattagga agtcagattc ttaaagagca tcctgttgaa
     3541 aatactcaat tgcaaaatga aaagctctat ctctattatc tccaaaatgg aagagacatg
     3601 tatgtggacc aagaattaga tattaatcgt ttaagtgatt atgatgtcga tcacattgtt
     3661 ccacaaagtt tccttaaaga cgattcaata gacaataagg tcttaacgcg ttctgataaa
     3721 aatcgtggta aatcggataa cgttccaagt gaagaagtag tcaaaaagat gaaaaactat
     3781 tggagacaac ttctaaacgc caagttaatc actcaacgta agtttgataa tttaacgaaa
     3841 gctgaacgtg gaggtttgag tgaacttgat aaagctggtt ttatcaaacg ccaattggtt
     3901 gaaactcgcc aaatcactaa gcatgtggca caaattttgg atagtcgcat gaatactaaa
     3961 tacgatgaaa atgataaact tattcgagag gttaaagtga ttaccttaaa atctaaatta
     4021 gtttctgact tccgaaaaga tttccaattc tataaagtac gtgagattaa caattaccat
     4081 catgcccatg atgcgtatct aaatgccgtc gttggaactg ctttgattaa gaaatatcca
     4141 aaacttgaat cggagtttgt ctatggtgat tataaagttt atgatgttcg taaaatgatt
     4201 gctaagtctg agcaagaaat aggcaaagca accgcaaaat atttctttta ctctaatatc
     4261 atgaacttct tcaaaacaga aattacactt gcaaatggag agattcgcaa acgccctcta
     4321 atcgaaacta atggggaaac tggagaaatt gtctgggata aagggcgaga ttttgccaca
     4381 gtgcgcaaag tattgtccat gccccaagtc aatattgtca agaaaacaga agtacagaca
     4441 ggcggattct ccaaggagtc aattttacca aaaagaaatt cggacaagct tattgctcgt
     4501 aaaaaagact gggatccaaa aaaatatggt ggttttgata gtccaacggt agcttattca
     4561 gtcctagtgg ttgctaaggt ggaaaaaggg aaatcgaaga agttaaaatc cgttaaagag
     4621 ttactaggga tcacaattat ggaaagaagt tcctttgaaa aaaatccgat tgacttttta
     4681 gaagctaaag gatataagga agttaaaaaa gacttaatca ttaaactacc taaatatagt
     4741 ctttttgagt tagaaaacgg tcgtaaacgg atgctggcta gtgccggaga attacaaaaa
     4801 ggaaatgagc tggctctgcc aagcaaatat gtgaattttt tatatttagc tagtcattat
     4861 gaaaagttga agggtagtcc agaagataac gaacaaaaac aattgtttgt ggagcagcat
     4921 aagcattatt tagatgagat tattgagcaa atcagtgaat tttctaagcg tgttatttta
     4981 gcagatgcca atttagataa agttcttagt gcatataaca aacatagaga caaaccaata
     5041 cgtgaacaag cagaaaatat tattcattta tttacgttga cgaatcttgg agctcccgct
     5101 gcttttaaat attttgatac aacaattgat cgtaaacgat atacgtctac aaaagaagtt
     5161 ttagatgcca ctcttatcca tcaatccatc actggtcttt atgaaacacg cattgatttg
     5221 agtcagctag gaggtgacgg gtcacctaag aaaaaacgaa aagttgagga tcctaaaaag
     5281 aaacgaaaag ttgataccgg ttctaaaggt gaagaactgt tcaccggtgt tgttccgatc
     5341 ctggttgaac tggatggtga tgttaacggc cacaaattct ctgttcgtgg tgaaggtgaa
     5401 ggtgatgcaa ccaacggtaa actgaccctg aaattcatct gcactaccgg taaactgccg
     5461 gttccatggc cgactctggt gactaccctg acctatggtg ttcagtgttt ttctcgttac
     5521 ccggatcaca tgaagcagca tgatttcttc aaatctgcaa tgccggaagg ttatgtacag
     5581 gagcgcacca tttctttcaa agacgatggc acctacaaaa cccgtgcaga ggttaaattt
     5641 gaaggtgata ctctggtgaa ccgtattgaa ctgaaaggca ttgatttcaa agaggacggc
     5701 aacatcctgg gccacaaact ggaatataac ttcaactccc ataacgttta catcaccgca
     5761 gacaaacaga agaacggtat caaagctaac ttcaaaattc gccataacgt tgaagacggt
     5821 agcgtacagc tggcggacca ctaccagcag aacactccga tcggtgatgg tccggttctg
     5881 ctgccggata accactacct gtccacccag tctgttctgt ccaaagaccc gaacgaaaag
     5941 cgcgaccaca tggtgctgct ggagttcgtt actgcagcag gtatcacgca cggcatggat
     6001 gagctctaca aataataaca ttggaagtgg ataacggatc cgcgatcgcg gcgcgccacc
     6061 tggtggccgg ccggtaccac gcgtgcgcgc tgatccggct gctaacaaag cccgaaagga
     6121 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     6181 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggatatc cacaggacgg
     6241 gtgtggtcgc catgatcgcg tagtcgatag tggctccaag tagcgaagcg agcaggactg
     6301 ggcggcggcc aaagcggtcg gacagtgctc cgagaacggg tgcgcataga aattgcatca
     6361 acgcatatag cgctagcagc acgccatagt gactggcgat gctgtcggaa tggacgatat
     6421 cccgcaagag gcccggcagt accggcataa ccaagcctat gcctacagca tccagggtga
     6481 cggtgccgag gatgacgatg agcgcattgt tagatttcat acacggtgcc tgactgcgtt
     6541 agcaatttaa ctgtgataaa ctaccgcatt aaagcttatc gatgataagc tgtcaaacat
     6601 gagaattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     6661 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     6721 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     6781 taaatgcttc aataacattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     6841 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     6901 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     6961 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     7021 tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc
     7081 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     7141 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     7201 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     7261 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     7321 gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc
     7381 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     7441 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     7501 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     7561 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     7621 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     7681 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     7741 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     7801 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     7861 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     7921 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     7981 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     8041 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     8101 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     8161 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     8221 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     8281 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     8341 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     8401 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     8461 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     8521 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     8581 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt
     8641 acgcatctgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc tgctctgatg
     8701 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     8761 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     8821 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     8881 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca
     8941 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     9001 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc
     9061 cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct
     9121 cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca
     9181 actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt
     9241 cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg
     9301 gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc
     9361 gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt
     9421 tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg
     9481 ggtcctcaac gacaggagca cgatcatgcg cacccgtggc caggacccaa cgctgcccga
     9541 gatgcgccgc gtgcggctgc tggagatggc ggacgcgatg gatatgttct gccaagggtt
     9601 ggtttgcgca ttcacagttc tccgcaagaa ttgattggct ccaattcttg gagtggtgaa
     9661 tccgttagcg aggtgccgcc ggcttccatt caggtcgagg tggcccggct ccatgcaccg
     9721 cgacgcaacg cggggaggca gacaaggtat agggcggcgc ctacaatcca tgccaacccg
     9781 ttccatgtgc tcgccgaggc ggcataaatc gccgtgacga tcagcggtcc aatgatcgaa
     9841 gttaggctgg taagagccgc gagcgatcct tgaagctgtc cctgatggtc gtcatctacc
     9901 tgcctggaca gcatggcctg caacgcgggc atcccgatgc cgccggaagc gagaagaatc
     9961 ataatgggga aggccatcca gcctcgcgtc gcgaacgcca gcaagacgta gcccagcgcg
    10021 tcggccgcca tgccggcgat aatggcctgc ttctcgccga aacgtttggt ggcgggacca
    10081 gtgacgaagg cttgagcgag ggcgtgcaag attccgaata ccgcaagcga caggccgatc
    10141 atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc
    10201 tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat agtcatgccc
    10261 cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg tcgacgctct
    10321 cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc
    10381 gccgccgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg
    10441 cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct
    10501 tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg
    10561 ccggccacga tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa ttaatacgac
    10621 tcactatagg gagaccacaa cggtttccct ctagtgccgg ctccggagag ctctttaatt
    10681 aagcggccgc cctgcaggac tcgagttcta gaaataattt tgtttaactt taagaaggag
    10741 atatacatat gaaatcttct caccatcacc atcaccatgg ttcttctatg aaaatcgaag
    10801 aaggtaaact ggtaatctgg attaacggcg ataaaggcta taac
//
