LOCUS       sequence-177717-       11066 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 11066)
  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 11066)
  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: 7xNLS-pMJ915v2-sfGFP
FEATURES             Location/Qualifiers
     source          1..11066
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     588..607
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     691..710
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        691..709
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             798..820
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             840..857
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             867..1967
                     /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     1941..1964
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             2022..2042
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2052..2072
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2082..2102
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2112..2132
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2142..2162
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2172..2192
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2202..2222
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2232..2252
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2280..6383
                     /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             2280..6383
                     /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             6390..6410
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6417..6437
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(7286..7304)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      7300..7347
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(7713..7718,7719..7735,7736..7741))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(7768..7786)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7854..7958
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7959..8027,8028..8819)
                     /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(8177..8196)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8990..9578
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9479..9498
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9732..9749
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    9764..9903
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(9829..9848)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9989..10011
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(10005..10196)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 aacgcgggca tcccgatgcc gccggaagcg agaagaatca taatggggaa ggccatccag
       61 cctcgcgtcg cgaacgccag caagacgtag cccagcgcgt cggccgccat gccggcgata
      121 atggcctgct tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg
      181 gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag
      241 cggtcctcgc cgaaaatgac ccagagcgct gccggcacct gtcctacgag ttgcatgata
      301 aagaagacag tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg
      361 actgggttga aggctctcaa gggcatcggt cgacgctctc ccttatgcga ctcctgcatt
      421 aggaagcagc ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca
      481 tgcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg
      541 aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg
      601 atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg
      661 tagaggatcg agatctcgat cccgcgaaat taatacgact cactataggg agaccacaac
      721 ggtttccctc tagtgccggc tccggagagc tctttaatta agcggccgcc ctgcaggact
      781 cgagttctag aaataatttt gtttaacttt aagaaggaga tatacatatg aaatcttctc
      841 accatcacca tcaccatggt tcttctatga aaatcgaaga aggtaaactg gtaatctgga
      901 ttaacggcga taaaggctat aacggtctcg ctgaagtcgg taagaaattc gagaaagata
      961 ccggaattaa agtcaccgtt gagcatccgg ataaactgga agagaaattc ccacaggttg
     1021 cggcaactgg cgatggccct gacattatct tctgggcaca cgaccgcttt ggtggctacg
     1081 ctcaatctgg cctgttggct gaaatcaccc cggacaaagc gttccaggac aagctgtatc
     1141 cgtttacctg ggatgccgta cgttacaacg gcaagctgat tgcttacccg atcgctgttg
     1201 aagcgttatc gctgatttat aacaaagatc tgctgccgaa cccgccaaaa acctgggaag
     1261 agatcccggc gctggataaa gaactgaaag cgaaaggtaa gagcgcgctg atgttcaacc
     1321 tgcaagaacc gtacttcacc tggccgctga ttgctgctga cgggggttat gcgttcaagt
     1381 atgaaaacgg caagtacgac attaaagacg tgggcgtgga taacgctggc gcgaaagcgg
     1441 gtctgacctt cctggttgac ctgattaaaa acaaacacat gaatgcagac accgattact
     1501 ccatcgcaga agctgccttt aataaaggcg aaacagcgat gaccatcaac ggcccgtggg
     1561 catggtccaa catcgacacc agcaaagtga attatggtgt aacggtactg ccgaccttca
     1621 agggtcaacc atccaaaccg ttcgttggcg tgctgagcgc aggtattaac gccgccagtc
     1681 cgaacaaaga gctggcaaaa gagttcctcg aaaactatct gctgactgat gaaggtctgg
     1741 aagcggttaa taaagacaaa ccgctgggtg ccgtagcgct gaagtcttac gaggaagagt
     1801 tggcgaaaga tccacgtatt gccgccacta tggaaaacgc ccagaaaggt gaaatcatgc
     1861 cgaacatccc gcagatgtcc gctttctggt atgccgtgcg tactgcggtg atcaacgccg
     1921 ccagcggtcg tcagactgtc gatgaagccc tgaaagacgc gcagactaat tcgagctcga
     1981 acaacaacaa caataacaat aacaacaacc tcgggatcga ggaaaacctg tacttccaat
     2041 ccaatgccac ccccaagaag aagcgaaagg tgggtgggtc cccaaagaag aaacggaaag
     2101 taggcggctc ccccaaaaag aagcgaaaag tagggggtag ccccaagaag aagcggaagg
     2161 taggtgggtc cccaaagaag aaacggaaag taggcggctc ccccaaaaag aagcgaaaag
     2221 tagggggtag ccccaagaag aagcggaagg taggtatcca cggagtccca gcagctacca
     2281 tggataagaa atactcaata ggcttagata tcggcacaaa tagcgtcgga tgggcggtga
     2341 tcactgatga atataaggtt ccgtctaaaa agttcaaggt tctgggaaat acagaccgcc
     2401 acagtatcaa aaaaaatctt ataggggctc ttttatttga cagtggagag acagcggaag
     2461 cgactcgtct caaacggaca gctcgtagaa ggtatacacg tcggaagaat cgtatttgtt
     2521 atctacagga gattttttca aatgagatgg cgaaagtaga tgatagtttc tttcatcgac
     2581 ttgaagagtc ttttttggtg gaagaagaca agaagcatga acgtcatcct atttttggaa
     2641 atatagtaga tgaagttgct tatcatgaga aatatccaac tatctatcat ctgcgaaaaa
     2701 aattggtaga ttctactgat aaagcggatt tgcgcttaat ctatttggcc ttagcgcata
     2761 tgattaagtt tcgtggtcat tttttgattg agggagattt aaatcctgat aatagtgatg
     2821 tggacaaact atttatccag ttggtacaaa cctacaatca attatttgaa gaaaacccta
     2881 ttaacgcaag tggagtagat gctaaagcga ttctttctgc acgattgagt aaatcaagac
     2941 gattagaaaa tctcattgct cagctccccg gtgagaagaa aaatggctta tttgggaatc
     3001 tcattgcttt gtcattgggt ttgaccccta attttaaatc aaattttgat ttggcagaag
     3061 atgctaaatt acagctttca aaagatactt acgatgatga tttagataat ttattggcgc
     3121 aaattggaga tcaatatgct gatttgtttt tggcagctaa gaatttatca gatgctattt
     3181 tactttcaga tatcctaaga gtaaatactg aaataactaa ggctccccta tcagcttcaa
     3241 tgattaaacg ctacgatgaa catcatcaag acttgactct tttaaaagct ttagttcgac
     3301 aacaacttcc agaaaagtat aaagaaatct tttttgatca atcaaaaaac ggatatgcag
     3361 gttatattga tgggggagct agccaagaag aattttataa atttatcaaa ccaattttag
     3421 aaaaaatgga tggtactgag gaattattgg tgaaactaaa tcgtgaagat ttgctgcgca
     3481 agcaacggac ctttgacaac ggctctattc cccatcaaat tcacttgggt gagctgcatg
     3541 ctattttgag aagacaagaa gacttttatc catttttaaa agacaatcgt gagaagattg
     3601 aaaaaatctt gacttttcga attccttatt atgttggtcc attggcgcgt ggcaatagtc
     3661 gttttgcatg gatgactcgg aagtctgaag aaacaattac cccatggaat tttgaagaag
     3721 ttgtcgataa aggtgcttca gctcaatcat ttattgaacg catgacaaac tttgataaaa
     3781 atcttccaaa tgaaaaagta ctaccaaaac atagtttgct ttatgagtat tttacggttt
     3841 ataacgaatt gacaaaggtc aaatatgtta ctgaaggaat gcgaaaacca gcatttcttt
     3901 caggtgaaca gaagaaagcc attgttgatt tactcttcaa aacaaatcga aaagtaaccg
     3961 ttaagcaatt aaaagaagat tatttcaaaa aaatagaatg ttttgatagt gttgaaattt
     4021 caggagttga agatagattt aatgcttcat taggtaccta ccatgatttg ctaaaaatta
     4081 ttaaagataa agattttttg gataatgaag aaaatgaaga tatcttagag gatattgttt
     4141 taacattgac cttatttgaa gatagggaga tgattgagga aagacttaaa acatatgctc
     4201 acctctttga tgataaggtg atgaaacagc ttaaacgtcg ccgttatact ggttggggac
     4261 gtttgtctcg aaaattgatt aatggtatta gggataagca atctggcaaa acaatattag
     4321 attttttgaa atcagatggt tttgccaatc gcaattttat gcagctgatc catgatgata
     4381 gtttgacatt taaagaagac attcaaaaag cacaagtgtc tggacaaggc gatagtttac
     4441 atgaacatat tgcaaattta gctggtagcc ctgctattaa aaaaggtatt ttacagactg
     4501 taaaagttgt tgatgaattg gtcaaagtaa tggggcggca taagccagaa aatatcgtta
     4561 ttgaaatggc acgtgaaaat cagacaactc aaaagggcca gaaaaattcg cgagagcgta
     4621 tgaaacgaat cgaagaaggt atcaaagaat taggaagtca gattcttaaa gagcatcctg
     4681 ttgaaaatac tcaattgcaa aatgaaaagc tctatctcta ttatctccaa aatggaagag
     4741 acatgtatgt ggaccaagaa ttagatatta atcgtttaag tgattatgat gtcgatcaca
     4801 ttgttccaca aagtttcctt aaagacgatt caatagacaa taaggtctta acgcgttctg
     4861 ataaaaatcg tggtaaatcg gataacgttc caagtgaaga agtagtcaaa aagatgaaaa
     4921 actattggag acaacttcta aacgccaagt taatcactca acgtaagttt gataatttaa
     4981 cgaaagctga acgtggaggt ttgagtgaac ttgataaagc tggttttatc aaacgccaat
     5041 tggttgaaac tcgccaaatc actaagcatg tggcacaaat tttggatagt cgcatgaata
     5101 ctaaatacga tgaaaatgat aaacttattc gagaggttaa agtgattacc ttaaaatcta
     5161 aattagtttc tgacttccga aaagatttcc aattctataa agtacgtgag attaacaatt
     5221 accatcatgc ccatgatgcg tatctaaatg ccgtcgttgg aactgctttg attaagaaat
     5281 atccaaaact tgaatcggag tttgtctatg gtgattataa agtttatgat gttcgtaaaa
     5341 tgattgctaa gtctgagcaa gaaataggca aagcaaccgc aaaatatttc ttttactcta
     5401 atatcatgaa cttcttcaaa acagaaatta cacttgcaaa tggagagatt cgcaaacgcc
     5461 ctctaatcga aactaatggg gaaactggag aaattgtctg ggataaaggg cgagattttg
     5521 ccacagtgcg caaagtattg tccatgcccc aagtcaatat tgtcaagaaa acagaagtac
     5581 agacaggcgg attctccaag gagtcaattt taccaaaaag aaattcggac aagcttattg
     5641 ctcgtaaaaa agactgggat ccaaaaaaat atggtggttt tgatagtcca acggtagctt
     5701 attcagtcct agtggttgct aaggtggaaa aagggaaatc gaagaagtta aaatccgtta
     5761 aagagttact agggatcaca attatggaaa gaagttcctt tgaaaaaaat ccgattgact
     5821 ttttagaagc taaaggatat aaggaagtta aaaaagactt aatcattaaa ctacctaaat
     5881 atagtctttt tgagttagaa aacggtcgta aacggatgct ggctagtgcc ggagaattac
     5941 aaaaaggaaa tgagctggct ctgccaagca aatatgtgaa ttttttatat ttagctagtc
     6001 attatgaaaa gttgaagggt agtccagaag ataacgaaca aaaacaattg tttgtggagc
     6061 agcataagca ttatttagat gagattattg agcaaatcag tgaattttct aagcgtgtta
     6121 ttttagcaga tgccaattta gataaagttc ttagtgcata taacaaacat agagacaaac
     6181 caatacgtga acaagcagaa aatattattc atttatttac gttgacgaat cttggagctc
     6241 ccgctgcttt taaatatttt gatacaacaa ttgatcgtaa acgatatacg tctacaaaag
     6301 aagttttaga tgccactctt atccatcaat ccatcactgg tctttatgaa acacgcattg
     6361 atttgagtca gctaggaggt gacgggtcac ctaagaaaaa acgaaaagtt gaggatccta
     6421 aaaagaaacg aaaagttgat accggttcta aaggtgaaga actgttcacc ggtgttgttc
     6481 cgatcctggt tgaactggat ggtgatgtta acggccacaa attctctgtt cgtggtgaag
     6541 gtgaaggtga tgcaaccaac ggtaaactga ccctgaaatt catctgcact accggtaaac
     6601 tgccggttcc atggccgact ctggtgacta ccctgaccta tggtgttcag tgtttttctc
     6661 gttacccgga tcacatgaag cagcatgatt tcttcaaatc tgcaatgccg gaaggttatg
     6721 tacaggagcg caccatttct ttcaaagacg atggcaccta caaaacccgt gcagaggtta
     6781 aatttgaagg tgatactctg gtgaaccgta ttgaactgaa aggcattgat ttcaaagagg
     6841 acggcaacat cctgggccac aaactggaat ataacttcaa ctcccataac gtttacatca
     6901 ccgcagacaa acagaagaac ggtatcaaag ctaacttcaa aattcgccat aacgttgaag
     6961 acggtagcgt acagctggcg gaccactacc agcagaacac tccgatcggt gatggtccgg
     7021 ttctgctgcc ggataaccac tacctgtcca cccagtctgt tctgtccaaa gacccgaacg
     7081 aaaagcgcga ccacatggtg ctgctggagt tcgttactgc agcaggtatc acgcacggca
     7141 tggatgagct ctacaaataa taacattgga agtggataac ggatccgcga tcgcggcgcg
     7201 ccacctggtg gccggccggt accacgcgtg cgcgctgatc cggctgctaa caaagcccga
     7261 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc
     7321 tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg atatccacag
     7381 gacgggtgtg gtcgccatga tcgcgtagtc gatagtggct ccaagtagcg aagcgagcag
     7441 gactgggcgg cggccaaagc ggtcggacag tgctccgaga acgggtgcgc atagaaattg
     7501 catcaacgca tatagcgcta gcagcacgcc atagtgactg gcgatgctgt cggaatggac
     7561 gatatcccgc aagaggcccg gcagtaccgg cataaccaag cctatgccta cagcatccag
     7621 ggtgacggtg ccgaggatga cgatgagcgc attgttagat ttcatacacg gtgcctgact
     7681 gcgttagcaa tttaactgtg ataaactacc gcattaaagc ttatcgatga taagctgtca
     7741 aacatgagaa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     7801 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     7861 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     7921 cctgataaat gcttcaataa cattgaaaaa ggaagagtat gagtattcaa catttccgtg
     7981 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     8041 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     8101 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     8161 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc
     8221 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     8281 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     8341 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     8401 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     8461 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt
     8521 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     8581 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     8641 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     8701 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     8761 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     8821 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     8881 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     8941 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     9001 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     9061 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     9121 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     9181 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     9241 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     9301 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     9361 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     9421 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     9481 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     9541 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     9601 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     9661 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     9721 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc
     9781 tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc
     9841 tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct
     9901 gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     9961 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
    10021 tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat
    10081 tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat
    10141 gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat
    10201 gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg
    10261 atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt
    10321 aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag
    10381 cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag
    10441 atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg
    10501 aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt
    10561 cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct
    10621 agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggccagga cccaacgctg
    10681 cccgagatgc gccgcgtgcg gctgctggag atggcggacg cgatggatat gttctgccaa
    10741 gggttggttt gcgcattcac agttctccgc aagaattgat tggctccaat tcttggagtg
    10801 gtgaatccgt tagcgaggtg ccgccggctt ccattcaggt cgaggtggcc cggctccatg
    10861 caccgcgacg caacgcgggg aggcagacaa ggtatagggc ggcgcctaca atccatgcca
    10921 acccgttcca tgtgctcgcc gaggcggcat aaatcgccgt gacgatcagc ggtccaatga
    10981 tcgaagttag gctggtaaga gccgcgagcg atccttgaag ctgtccctga tggtcgtcat
    11041 ctacctgcct ggacagcatg gcctgc
//
