LOCUS       sequence-313969-        9582 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro transcription of SpCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9582)
  AUTHORS   Pelletier S, Gingras S, Green DR
  TITLE     Mouse genome engineering via CRISPR-Cas9 for study of immune 
            function.
  JOURNAL   Immunity. 2015 Jan 20;42(1):18-27. doi: 
            10.1016/j.immuni.2015.01.004.
  PUBMED    25607456
REFERENCE   2  (bases 1 to 9582)
  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"; doi: 
            "10.1016/j.immuni.2015.01"; journalName: "Immunity"; date: 
            "2015-01-20- 20"; volume: "42"; issue: "1"; pages: "18-27"
COMMENT     Sequence Label: pCDNA3.3TOPO-T7-hCas9
FEATURES             Location/Qualifiers
     source          1..9582
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        384..587
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     538..558
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     584..608
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     683..702
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        683..701
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      725..734
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             731..4834
                     /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             731..4834
                     /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             731..4834
                     /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             4847..4867
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4922..4941)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4966..5014
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5216..5644
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5303..5322)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5513..5534
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5653..5673)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5658..5987
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5838..5973
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5900..5919
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6054..6848
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6108..6127)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6718..6737
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7024..7145
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7061..7080)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7115..7134
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7194..7210)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7194..7210)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7207..7229)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7218..7234
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(7242..7248,7249..7266,7267..7272))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7287..7308
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7425..7442)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7596..8184)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7676..7695)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8355..9146,9147..9215))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8978..8997
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9216..9320)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9395..9414)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      421 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      481 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      541 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc
      601 gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga caccgggacc
      661 gatccagcct ccggactcta gataatacga ctcactatag ggagatctag aggatcgaac
      721 ccttgccacc atggacaaga agtactccat tgggctcgat atcggcacaa acagcgtcgg
      781 ctgggccgtc attacggacg agtacaaggt gccgagcaaa aaattcaaag ttctgggcaa
      841 taccgatcgc cacagcataa agaagaacct cattggcgcc ctcctgttcg actccgggga
      901 gacggccgaa gccacgcggc tcaaaagaac agcacggcgc agatataccc gcagaaagaa
      961 tcggatctgc tacctgcagg agatctttag taatgagatg gctaaggtgg atgactcttt
     1021 cttccatagg ctggaggagt cctttttggt ggaggaggat aaaaagcacg agcgccaccc
     1081 aatctttggc aatatcgtgg acgaggtggc gtaccatgaa aagtacccaa ccatatatca
     1141 tctgaggaag aagcttgtag acagtactga taaggctgac ttgcggttga tctatctcgc
     1201 gctggcgcat atgatcaaat ttcggggaca cttcctcatc gagggggacc tgaacccaga
     1261 caacagcgat gtcgacaaac tctttatcca actggttcag acttacaatc agcttttcga
     1321 agagaacccg atcaacgcat ccggagttga cgccaaagca atcctgagcg ctaggctgtc
     1381 caaatcccgg cggctcgaaa acctcatcgc acagctccct ggggagaaga agaacggcct
     1441 gtttggtaat cttatcgccc tgtcactcgg gctgaccccc aactttaaat ctaacttcga
     1501 cctggccgaa gatgccaagc ttcaactgag caaagacacc tacgatgatg atctcgacaa
     1561 tctgctggcc cagatcggcg accagtacgc agaccttttt ttggcggcaa agaacctgtc
     1621 agacgccatt ctgctgagtg atattctgcg agtgaacacg gagatcacca aagctccgct
     1681 gagcgctagt atgatcaagc gctatgatga gcaccaccaa gacttgactt tgctgaaggc
     1741 ccttgtcaga cagcaactgc ctgagaagta caaggaaatt ttcttcgatc agtctaaaaa
     1801 tggctacgcc ggatacattg acggcggagc aagccaggag gaattttaca aatttattaa
     1861 gcccatcttg gaaaaaatgg acggcaccga ggagctgctg gtaaagctta acagagaaga
     1921 tctgttgcgc aaacagcgca ctttcgacaa tggaagcatc ccccaccaga ttcacctggg
     1981 cgaactgcac gctatcctca ggcggcaaga ggatttctac ccctttttga aagataacag
     2041 ggaaaagatt gagaaaatcc tcacatttcg gataccctac tatgtaggcc ccctcgcccg
     2101 gggaaattcc agattcgcgt ggatgactcg caaatcagaa gagaccatca ctccctggaa
     2161 cttcgaggaa gtcgtggata agggggcctc tgcccagtcc ttcatcgaaa ggatgactaa
     2221 ctttgataaa aatctgccta acgaaaaggt gcttcctaaa cactctctgc tgtacgagta
     2281 cttcacagtt tataacgagc tcaccaaggt caaatacgtc acagaaggga tgagaaagcc
     2341 agcattcctg tctggagagc agaagaaagc tatcgtggac ctcctcttca agacgaaccg
     2401 gaaagttacc gtgaaacagc tcaaagaaga ctatttcaaa aagattgaat gtttcgactc
     2461 tgttgaaatc agcggagtgg aggatcgctt caacgcatcc ctgggaacgt atcacgatct
     2521 cctgaaaatc attaaagaca aggacttcct ggacaatgag gagaacgagg acattcttga
     2581 ggacattgtc ctcaccctta cgttgtttga agatagggag atgattgaag aacgcttgaa
     2641 aacttacgct catctcttcg acgacaaagt catgaaacag ctcaagaggc gccgatatac
     2701 aggatggggg cggctgtcaa gaaaactgat caatgggatc cgagacaagc agagtggaaa
     2761 gacaatcctg gattttctta agtccgatgg atttgccaac cggaacttca tgcagttgat
     2821 ccatgatgac tctctcacct ttaaggagga catccagaaa gcacaagttt ctggccaggg
     2881 ggacagtctt cacgagcaca tcgctaatct tgcaggtagc ccagctatca aaaagggaat
     2941 actgcagacc gttaaggtcg tggatgaact cgtcaaagta atgggaaggc ataagcccga
     3001 gaatatcgtt atcgagatgg cccgagagaa ccaaactacc cagaagggac agaagaacag
     3061 tagggaaagg atgaagagga ttgaagaggg tataaaagaa ctggggtccc aaatccttaa
     3121 ggaacaccca gttgaaaaca cccagcttca gaatgagaag ctctacctgt actacctgca
     3181 gaacggcagg gacatgtacg tggatcagga actggacatc aatcggctct ccgactacga
     3241 cgtggatcat atcgtgcccc agtcttttct caaagatgat tctattgata ataaagtgtt
     3301 gacaagatcc gataaaaata gagggaagag tgataacgtc ccctcagaag aagttgtcaa
     3361 gaaaatgaaa aattattggc ggcagctgct gaacgccaaa ctgatcacac aacggaagtt
     3421 cgataatctg actaaggctg aacgaggtgg cctgtctgag ttggataaag ccggcttcat
     3481 caaaaggcag cttgttgaga cacgccagat caccaagcac gtggcccaaa ttctcgattc
     3541 acgcatgaac accaagtacg atgaaaatga caaactgatt cgagaggtga aagttattac
     3601 tctgaagtct aagctggtct cagatttcag aaaggacttt cagttttata aggtgagaga
     3661 gatcaacaat taccaccatg cgcatgatgc ctacctgaat gcagtggtag gcactgcact
     3721 tatcaaaaaa tatcccaagc ttgaatctga atttgtttac ggagactata aagtgtacga
     3781 tgttaggaaa atgatcgcaa agtctgagca ggaaataggc aaggccaccg ctaagtactt
     3841 cttttacagc aatattatga attttttcaa gaccgagatt acactggcca atggagagat
     3901 tcggaagcga ccacttatcg aaacaaacgg agaaacagga gaaatcgtgt gggacaaggg
     3961 tagggatttc gcgacagtcc ggaaggtcct gtccatgccg caggtgaaca tcgttaaaaa
     4021 gaccgaagta cagaccggag gcttctccaa ggaaagtatc ctcccgaaaa ggaacagcga
     4081 caagctgatc gcacgcaaaa aagattggga ccccaagaaa tacggcggat tcgattctcc
     4141 tacagtcgct tacagtgtac tggttgtggc caaagtggag aaagggaagt ctaaaaaact
     4201 caaaagcgtc aaggaactgc tgggcatcac aatcatggag cgatcaagct tcgaaaaaaa
     4261 ccccatcgac tttctcgagg cgaaaggata taaagaggtc aaaaaagacc tcatcattaa
     4321 gcttcccaag tactctctct ttgagcttga aaacggccgg aaacgaatgc tcgctagtgc
     4381 gggcgagctg cagaaaggta acgagctggc actgccctct aaatacgtta atttcttgta
     4441 tctggccagc cactatgaaa agctcaaagg gtctcccgaa gataatgagc agaagcagct
     4501 gttcgtggaa caacacaaac actaccttga tgagatcatc gagcaaataa gcgaattctc
     4561 caaaagagtg atcctcgccg acgctaacct cgataaggtg ctttctgctt acaataagca
     4621 cagggataag cccatcaggg agcaggcaga aaacattatc cacttgttta ctctgaccaa
     4681 cttgggcgcg cctgcagcct tcaagtactt cgacaccacc atagacagaa agcggtacac
     4741 ctctacaaag gaggtcctgg acgccacact gattcatcag tcaattacgg ggctctatga
     4801 aacaagaatc gacctctctc agctcggtgg agacagcagg gctgacccca agaagaagag
     4861 gaaggtgtga aagggttcga tccctaccgg ttagtaatga gtttaaacgg gggaggctaa
     4921 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     4981 agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     5041 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     5101 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     5161 gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta tccccacgcg
     5221 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     5281 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     5341 gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg atttagtgct
     5401 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     5461 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     5521 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     5581 attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     5641 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     5701 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     5761 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     5821 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     5881 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     5941 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     6001 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     6061 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     6121 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     6181 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     6241 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     6301 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     6361 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     6421 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     6481 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     6541 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     6601 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     6661 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     6721 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     6781 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     6841 tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca
     6901 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     6961 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     7021 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     7081 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     7141 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     7201 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     7261 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     7321 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     7381 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     7441 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     7501 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     7561 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     7621 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     7681 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     7741 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc
     7801 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     7861 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     7921 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     7981 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     8041 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     8101 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     8161 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     8221 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     8281 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     8341 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     8401 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     8461 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     8521 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     8581 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     8641 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     8701 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     8761 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     8821 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     8881 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     8941 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     9001 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     9061 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     9121 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     9181 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     9241 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     9301 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
     9361 ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat gccgcatagt
     9421 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
     9481 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta
     9541 ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gc
//
