LOCUS       sequence-241825-       11400 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cas9-5xFlag-TdT in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11400)
  AUTHORS   Loveless TB, Grotts JH, Schechter MW, Forouzmand E, Carlson CK, 
            Agahi BS, Liang G, Ficht M, Liu B, Xie X, Liu CC
  TITLE     Lineage tracing and analog recording in mammalian cells by 
            single-site DNA writing.
  JOURNAL   Nat Chem Biol. 2021 Jun;17(6):739-747. doi: 
            10.1038/s41589-021-00769-8. Epub 2021 Mar 22.
  PUBMED    33753928
REFERENCE   2  (bases 1 to 11400)
  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: 
            "10.1038/s41589-021-00769-8"; journalName: "Nat Chem Biol"; date: 
            "2021-06"; volume: "17"; issue: "6"; pages: "739-747"
COMMENT     Sequence Label: pTBL227 NheI-Cas9-KpnI-5xFlag-FseI-TdT-NotI
FEATURES             Location/Qualifiers
     source          1..11400
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      911..920
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             935..5038
                     /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             935..5038
                     /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             935..5038
                     /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             5051..5071
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5078..5209
                     /codon_start=1
                     /product="five tandem FLAG(R) epitope tags"
                     /label=5xFLAG
                     /translation="DYKDDDDKDYKDDDDKLMDYKDDDDKDYKDDDDKLMDYKDDDDK"
     primer_bind     complement(6838..6855)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6844..7068
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7114..7542
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7201..7220)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7411..7432
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7551..7571)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7556..7885
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7736..7871
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7798..7817
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    9076..9197
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9113..9132)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9167..9186
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9246..9262)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9246..9262)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9259..9281)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9270..9286
                     /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(9294..9300,9301..9318,9319..9324))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9339..9360
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9477..9494)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9648..10233)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9728..9747)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10404..11195,11196..11264))
                     /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     11027..11046
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11265..11369)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gctaccggtc gccaccatgg gccctagtgg cccaatggac aagaagtact ccattgggct
      961 cgatatcggc acaaacagcg tcggctgggc cgtcattacg gacgagtaca aggtgccgag
     1021 caaaaaattc aaagttctgg gcaataccga tcgccacagc ataaagaaga acctcattgg
     1081 cgccctcctg ttcgactccg gggagacggc cgaagccacg cggctcaaaa gaacagcacg
     1141 gcgcagatat acccgcagaa agaatcggat ctgctacctg caggagatct ttagtaatga
     1201 gatggctaag gtggatgact ctttcttcca taggctggag gagtcctttt tggtggagga
     1261 ggataaaaag cacgagcgcc acccaatctt tggcaatatc gtggacgagg tggcgtacca
     1321 tgaaaagtac ccaaccatat atcatctgag gaagaagctt gtagacagta ctgataaggc
     1381 tgacttgcgg ttgatctatc tcgcgctggc gcatatgatc aaatttcggg gacacttcct
     1441 catcgagggg gacctgaacc cagacaacag cgatgtcgac aaactcttta tccaactggt
     1501 tcagacttac aatcagcttt tcgaagagaa cccgatcaac gcatccggag ttgacgccaa
     1561 agcaatcctg agcgctaggc tgtccaaatc ccggcggctc gaaaacctca tcgcacagct
     1621 ccctggggag aagaagaacg gcctgtttgg taatcttatc gccctgtcac tcgggctgac
     1681 ccccaacttt aaatctaact tcgacctggc cgaagatgcc aagcttcaac tgagcaaaga
     1741 cacctacgat gatgatctcg acaatctgct ggcccagatc ggcgaccagt acgcagacct
     1801 ttttttggcg gcaaagaacc tgtcagacgc cattctgctg agtgatattc tgcgagtgaa
     1861 cacggagatc accaaagctc cgctgagcgc tagtatgatc aagcgctatg atgagcacca
     1921 ccaagacttg actttgctga aggcccttgt cagacagcaa ctgcctgaga agtacaagga
     1981 aattttcttc gatcagtcta aaaatggcta cgccggatac attgacggcg gagcaagcca
     2041 ggaggaattt tacaaattta ttaagcccat cttggaaaaa atggacggca ccgaggagct
     2101 gctggtaaag cttaacagag aagatctgtt gcgcaaacag cgcactttcg acaatggaag
     2161 catcccccac cagattcacc tgggcgaact gcacgctatc ctcaggcggc aagaggattt
     2221 ctaccccttt ttgaaagata acagggaaaa gattgagaaa atcctcacat ttcggatacc
     2281 ctactatgta ggccccctcg cccggggaaa ttccagattc gcgtggatga ctcgcaaatc
     2341 agaagagacc atcactccct ggaacttcga ggaagtcgtg gataaggggg cctctgccca
     2401 gtccttcatc gaaaggatga ctaactttga taaaaatctg cctaacgaaa aggtgcttcc
     2461 taaacactct ctgctgtacg agtacttcac agtttataac gagctcacca aggtcaaata
     2521 cgtcacagaa gggatgagaa agccagcatt cctgtctgga gagcagaaga aagctatcgt
     2581 ggacctcctc ttcaagacga accggaaagt taccgtgaaa cagctcaaag aagactattt
     2641 caaaaagatt gaatgtttcg actctgttga aatcagcgga gtggaggatc gcttcaacgc
     2701 atccctggga acgtatcacg atctcctgaa aatcattaaa gacaaggact tcctggacaa
     2761 tgaggagaac gaggacattc ttgaggacat tgtcctcacc cttacgttgt ttgaagatag
     2821 ggagatgatt gaagaacgct tgaaaactta cgctcatctc ttcgacgaca aagtcatgaa
     2881 acagctcaag aggcgccgat atacaggatg ggggcggctg tcaagaaaac tgatcaatgg
     2941 gatccgagac aagcagagtg gaaagacaat cctggatttt cttaagtccg atggatttgc
     3001 caaccggaac ttcatgcagt tgatccatga tgactctctc acctttaagg aggacatcca
     3061 gaaagcacaa gtttctggcc agggggacag tcttcacgag cacatcgcta atcttgcagg
     3121 tagcccagct atcaaaaagg gaatactgca gaccgttaag gtcgtggatg aactcgtcaa
     3181 agtaatggga aggcataagc ccgagaatat cgttatcgag atggcccgag agaaccaaac
     3241 tacccagaag ggacagaaga acagtaggga aaggatgaag aggattgaag agggtataaa
     3301 agaactgggg tcccaaatcc ttaaggaaca cccagttgaa aacacccagc ttcagaatga
     3361 gaagctctac ctgtactacc tgcagaacgg cagggacatg tacgtggatc aggaactgga
     3421 catcaatcgg ctctccgact acgacgtgga tcatatcgtg ccccagtctt ttctcaaaga
     3481 tgattctatt gataataaag tgttgacaag atccgataaa aatagaggga agagtgataa
     3541 cgtcccctca gaagaagttg tcaagaaaat gaaaaattat tggcggcagc tgctgaacgc
     3601 caaactgatc acacaacgga agttcgataa tctgactaag gctgaacgag gtggcctgtc
     3661 tgagttggat aaagccggct tcatcaaaag gcagcttgtt gagacacgcc agatcaccaa
     3721 gcacgtggcc caaattctcg attcacgcat gaacaccaag tacgatgaaa atgacaaact
     3781 gattcgagag gtgaaagtta ttactctgaa gtctaagctg gtctcagatt tcagaaagga
     3841 ctttcagttt tataaggtga gagagatcaa caattaccac catgcgcatg atgcctacct
     3901 gaatgcagtg gtaggcactg cacttatcaa aaaatatccc aagcttgaat ctgaatttgt
     3961 ttacggagac tataaagtgt acgatgttag gaaaatgatc gcaaagtctg agcaggaaat
     4021 aggcaaggcc accgctaagt acttctttta cagcaatatt atgaattttt tcaagaccga
     4081 gattacactg gccaatggag agattcggaa gcgaccactt atcgaaacaa acggagaaac
     4141 aggagaaatc gtgtgggaca agggtaggga tttcgcgaca gtccggaagg tcctgtccat
     4201 gccgcaggtg aacatcgtta aaaagaccga agtacagacc ggaggcttct ccaaggaaag
     4261 tatcctcccg aaaaggaaca gcgacaagct gatcgcacgc aaaaaagatt gggaccccaa
     4321 gaaatacggc ggattcgatt ctcctacagt cgcttacagt gtactggttg tggccaaagt
     4381 ggagaaaggg aagtctaaaa aactcaaaag cgtcaaggaa ctgctgggca tcacaatcat
     4441 ggagcgatca agcttcgaaa aaaaccccat cgactttctc gaggcgaaag gatataaaga
     4501 ggtcaaaaaa gacctcatca ttaagcttcc caagtactct ctctttgagc ttgaaaacgg
     4561 ccggaaacga atgctcgcta gtgcgggcga gctgcagaaa ggtaacgagc tggcactgcc
     4621 ctctaaatac gttaatttct tgtatctggc cagccactat gaaaagctca aagggtctcc
     4681 cgaagataat gagcagaagc agctgttcgt ggaacaacac aaacactacc ttgatgagat
     4741 catcgagcaa ataagcgaat tctccaaaag agtgatcctc gccgacgcta acctcgataa
     4801 ggtgctttct gcttacaata agcacaggga taagcccatc agggagcagg cagaaaacat
     4861 tatccacttg tttactctga ccaacttggg cgcgcctgca gccttcaagt acttcgacac
     4921 caccatagac agaaagcggt acacctctac aaaggaggtc ctggacgcca cactgattca
     4981 tcagtcaatt acggggctct atgaaacaag aatcgacctc tctcagctcg gtggagacag
     5041 cagggctgac cccaagaaga agaggaaggt gggtaccgac tacaaggacg acgatgacaa
     5101 ggattacaaa gatgacgacg ataagcttat ggactacaag gacgacgatg acaaggatta
     5161 caaagatgac gacgataagc ttatggacta caaggacgac gatgacaagc atatgggcgc
     5221 gccacttcta aataagcgaa tttcttatga tttaggccgg ccaatggatc caccacgagc
     5281 gtcccacttg agccctcgga agaagagacc ccggcagacg ggtgccttga tggcctcctc
     5341 tcctcaagac atcaaatttc aagatttggt cgtcttcatt ttggagaaga aaatgggaac
     5401 cacccgcaga gcgttcctca tggagctggc ccgcaggaaa gggttcaggg ttgaaaatga
     5461 gctcagtgat tctgtcaccc acattgtagc agagaacaac tcgggttcgg atgttctgga
     5521 gtggcttcaa gcacagaaag tacaagtcag ctcacaacca gagctcctcg atgtctcctg
     5581 gctgatcgaa tgcataggag cagggaaacc ggtggaaatg acaggaaaac accagcttgt
     5641 tgtgagaaga gactattcag atagcaccaa cccaggcccc ccgaagactc caccaattgc
     5701 tgtacaaaag atctcccagt atgcgtgtca gagaagaacc actttaaaca actgtaacca
     5761 gatattcacg gatgcctttg atatactggc tgaaaactgt gagtttagag aaaatgaaga
     5821 ctcctgtgtg acatttatga gagcagcttc tgtattgaaa tctctgccat tcacaatcat
     5881 cagtatgaag gacacagaag gaattccctg cctggggtcc aaggtgaagg gtatcataga
     5941 ggagattatt gaagatggag aaagttctga agttaaagct gtgttaaatg atgaacgata
     6001 tcaatccttc aaactcttta cttctgtatt tggagtgggg ctgaagactt ctgagaagtg
     6061 gttcaggatg ggtttcagaa ctctgagtaa agtaaggtcg gacaaaagcc tgaaatttac
     6121 acgaatgcag aaagcaggat ttctgtatta tgaagacctt gtcagctgtg tgaccagggc
     6181 agaagcagag gccgtcagtg tgctggttaa agaggctgtc tgggcatttc ttccggatgc
     6241 tttcgtcacc atgacaggag ggttccggag gggtaagaag atggggcatg atgtagattt
     6301 tttaattacc agcccaggat caacagagga tgaagagcaa cttttacaga aagtgatgaa
     6361 cttatgggaa aagaagggat tacttttata ttatgacctt gtggagtcaa catttgaaaa
     6421 gctcaggttg cctagcagga aggttgatgc tttggatcat tttcaaaagt gctttctgat
     6481 tttcaaattg cctcgtcaaa gagtggacag tgaccagtcc agctggcagg aaggaaagac
     6541 ctggaaggcc atccgtgtgg atttagttct gtgcccctac gagcgtcgtg cctttgccct
     6601 gttgggatgg actggctccc ggcagtttga gagagacctc cggcgctatg ccacacatga
     6661 gcggaagatg attctggata accatgcttt atatgacaag accaagagga tattcctcaa
     6721 agcagaaagt gaagaagaaa tttttgcgca tctgggattg gattatattg aaccgtggga
     6781 aagaaatgcc taggcggccg ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct
     6841 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     6901 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     6961 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     7021 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     7081 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     7141 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     7201 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     7261 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     7321 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     7381 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     7441 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     7501 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     7561 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     7621 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     7681 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     7741 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     7801 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     7861 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     7921 tgatcaagag acaggatgag gatcgtttcg catgcagctc tcggagggcg aagaatctcg
     7981 tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga
     8041 tggtttctac aaagatcgtt atgtttatcg gcactttgca tcggccgcgc tcccgattcc
     8101 ggaagtgctt gacattgggg aatttagcga gagcctgacc tattgcatct cccgccgtgc
     8161 acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcaaccggt
     8221 cgcggaggcc atggatgcga tcgctgcggc cgatcttagc cagacgagcg ggttcggccc
     8281 attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc
     8341 tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc
     8401 gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt
     8461 gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat
     8521 tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg
     8581 gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga
     8641 gcttgcagga tcgccgcggc tccgggcgta tatgctccgc attggtcttg accaactcta
     8701 tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc
     8761 aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc
     8821 cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac
     8881 tcgtccgagg gcaaaggaat aggcgggact ctggggttcg aaatgaccga ccaagcgacg
     8941 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     9001 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     9061 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     9121 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     9181 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     9241 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     9301 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     9361 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     9421 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     9481 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     9541 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     9601 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     9661 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     9721 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     9781 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     9841 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     9901 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     9961 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
    10021 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
    10081 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
    10141 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtttttt tgtttgcaag
    10201 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
    10261 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
    10321 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
    10381 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
    10441 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
    10501 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
    10561 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
    10621 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
    10681 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
    10741 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
    10801 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
    10861 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
    10921 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
    10981 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
    11041 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
    11101 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
    11161 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
    11221 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
    11281 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
    11341 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
