LOCUS       sequence-380004-       10289 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express TadCBEd-V106W in mammalian cells with BlastR selection, Tol2
            integration.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10289)
  AUTHORS   Neugebauer ME, Hsu A, Arbab M, Krasnow NA, McElroy AN, Pandey S, 
            Doman JL, Huang TP, Raguram A, Banskota S, Newby GA, Tolar J, Osborn
            MJ, Liu DR
  TITLE     Evolution of an adenine base editor into a small, efficient cytosine
            base editor with low off-target activity.
  JOURNAL   Nat Biotechnol. 2022 Nov 10. pii: 10.1038/s41587-022-01533-6. doi: 
            10.1038/s41587-022-01533-6.
  PUBMED    36357719
REFERENCE   2  (bases 1 to 10289)
  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. 2022 Nov 10. pii: 10.1038/s41587-022-01533-6. doi: 
            10.1038/s41587-022-01533-6."
COMMENT     Sequence Label: p2T-cmv-SpCas9-TadCBEd-V106W-BlastR
FEATURES             Location/Qualifiers
     source          1..10289
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     8..27
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8..26
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             76..96
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(694..717,718..720,721..4794)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             694..4794
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4825..5073
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5104..5352
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5395..5415
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5424..5441
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5464..5481)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5470..5694
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5765..5781)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5765..5781)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5778..5800)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5789..5805
                     /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(5813..5819,5820..5837,5838..5843))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5858..5879
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     polyA_signal    5996..6117
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6007..6026)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6061..6080
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(6275..6673)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     promoter        complement(6692..6739)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(6787..7116)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6801..6936
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6855..6874)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7101..7121
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     misc_feature    join(7240..7427,7428..7439)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     rep_origin      complement(7500..8088)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7580..7599)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8259..9050,9051..9119))
                     /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     8882..8901
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9120..9224)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9303..9322)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     misc_feature    join(9514..9525,9526..9663)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     enhancer        9672..10051
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        10052..10255
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     10206..10226
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 cggccgctaa tacgactcac tatagggaga gccgccacca tgaaacggac agccgacgga
       61 agcgagttcg agtcaccaaa gaagaagcgg aaagtcagtt ctgaggtgga gttttcccac
      121 gagtactgga tgagacatgc cctgaccctg gccaagaggg cacgggatga gaggaaggcg
      181 cctgtgggag ccgtgctggt gctgaacaat agagtgatcg gcgagggctg gaacagagcc
      241 atcggcctgc acgacccaac agcccatgcc gaaattatag ccctgagaca gggcggcctg
      301 gtcatgcaga actacagact gattgacgcc accctgtacg tgacattcga gccttgcgtg
      361 atgtgcgccg gcgccatgat caactctagg atcggccgcg tggtgtttgg ctggaggaac
      421 tcaaaaagag gcgccgcagg ctccctgatg aacgtgctga actaccccgg aatgaatcac
      481 cgcgtcgaaa ttaccgaggg aatcctggca gatgaatgtg ccgccctgct gtgcgatttc
      541 tatcggatgc ctagacaggt gttcaatgct cagaagaagg cccagagctc catcaactct
      601 ggcggatcta gcggaggatc ctctggcagc gagacaccag gaacaagcga gtcagcaaca
      661 ccagagagca gtggcggcag cagcggcggc agcgacaaga agtacagcat cggcctggcc
      721 atcggcacca actctgtggg ctgggccgtg atcaccgacg agtacaaggt gcccagcaag
      781 aaattcaagg tgctgggcaa caccgaccgg cacagcatca agaagaacct gatcggagcc
      841 ctgctgttcg acagcggcga aacagccgag gccacccggc tgaagagaac cgccagaaga
      901 agatacacca gacggaagaa ccggatctgc tatctgcaag agatcttcag caacgagatg
      961 gccaaggtgg acgacagctt cttccacaga ctggaagagt ccttcctggt ggaagaggat
     1021 aagaagcacg agcggcaccc catcttcggc aacatcgtgg acgaggtggc ctaccacgag
     1081 aagtacccca ccatctacca cctgagaaag aaactggtgg acagcaccga caaggccgac
     1141 ctgcggctga tctatctggc cctggcccac atgatcaagt tccggggcca cttcctgatc
     1201 gagggcgacc tgaaccccga caacagcgac gtggacaagc tgttcatcca gctggtgcag
     1261 acctacaacc agctgttcga ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc
     1321 atcctgtctg ccagactgag caagagcaga cggctggaaa atctgatcgc ccagctgccc
     1381 ggcgagaaga agaatggcct gttcggaaac ctgattgccc tgagcctggg cctgaccccc
     1441 aacttcaaga gcaacttcga cctggccgag gatgccaaac tgcagctgag caaggacacc
     1501 tacgacgacg acctggacaa cctgctggcc cagatcggcg accagtacgc cgacctgttt
     1561 ctggccgcca agaacctgtc cgacgccatc ctgctgagcg acatcctgag agtgaacacc
     1621 gagatcacca aggcccccct gagcgcctct atgatcaaga gatacgacga gcaccaccag
     1681 gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta caaagagatt
     1741 ttcttcgacc agagcaagaa cggctacgcc ggctacattg acggcggagc cagccaggaa
     1801 gagttctaca agttcatcaa gcccatcctg gaaaagatgg acggcaccga ggaactgctc
     1861 gtgaagctga acagagagga cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc
     1921 ccccaccaga tccacctggg agagctgcac gccattctgc ggcggcagga agatttttac
     1981 ccattcctga aggacaaccg ggaaaagatc gagaagatcc tgaccttccg catcccctac
     2041 tacgtgggcc ctctggccag gggaaacagc agattcgcct ggatgaccag aaagagcgag
     2101 gaaaccatca ccccctggaa cttcgaggaa gtggtggaca agggcgcttc cgcccagagc
     2161 ttcatcgagc ggatgaccaa cttcgataag aacctgccca acgagaaggt gctgcccaag
     2221 cacagcctgc tgtacgagta cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg
     2281 accgagggaa tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc catcgtggac
     2341 ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag
     2401 aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc
     2461 ctgggcacat accacgatct gctgaaaatt atcaaggaca aggacttcct ggacaatgag
     2521 gaaaacgagg acattctgga agatatcgtg ctgaccctga cactgtttga ggacagagag
     2581 atgatcgagg aacggctgaa aacctatgcc cacctgttcg acgacaaagt gatgaagcag
     2641 ctgaagcggc ggagatacac cggctggggc aggctgagcc ggaagctgat caacggcatc
     2701 cgggacaagc agtccggcaa gacaatcctg gatttcctga agtccgacgg cttcgccaac
     2761 agaaacttca tgcagctgat ccacgacgac agcctgacct ttaaagagga catccagaaa
     2821 gcccaggtgt ccggccaggg cgatagcctg cacgagcaca ttgccaatct ggccggcagc
     2881 cccgccatta agaagggcat cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg
     2941 atgggccggc acaagcccga gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc
     3001 cagaagggac agaagaacag ccgcgagaga atgaagcgga tcgaagaggg catcaaagag
     3061 ctgggcagcc agatcctgaa agaacacccc gtggaaaaca cccagctgca gaacgagaag
     3121 ctgtacctgt actacctgca gaatgggcgg gatatgtacg tggaccagga actggacatc
     3181 aaccggctgt ccgactacga tgtggaccat atcgtgcctc agagctttct gaaggacgac
     3241 tccatcgaca acaaggtgct gaccagaagc gacaagaacc ggggcaagag cgacaacgtg
     3301 ccctccgaag aggtcgtgaa gaagatgaag aactactggc ggcagctgct gaacgccaag
     3361 ctgattaccc agagaaagtt cgacaatctg accaaggccg agagaggcgg cctgagcgaa
     3421 ctggataagg ccggcttcat caagagacag ctggtggaaa cccggcagat cacaaagcac
     3481 gtggcacaga tcctggactc ccggatgaac actaagtacg acgagaatga caagctgatc
     3541 cgggaagtga aagtgatcac cctgaagtcc aagctggtgt ccgatttccg gaaggatttc
     3601 cagttttaca aagtgcgcga gatcaacaac taccaccacg cccacgacgc ctacctgaac
     3661 gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc tggaaagcga gttcgtgtac
     3721 ggcgactaca aggtgtacga cgtgcggaag atgatcgcca agagcgagca ggaaatcggc
     3781 aaggctaccg ccaagtactt cttctacagc aacatcatga actttttcaa gaccgagatt
     3841 accctggcca acggcgagat ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg
     3901 gagatcgtgt gggataaggg ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc
     3961 caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa agagtctatc
     4021 ctgcccaaga ggaacagcga taagctgatc gccagaaaga aggactggga ccctaagaag
     4081 tacggcggct tcgacagccc caccgtggcc tattctgtgc tggtggtggc caaagtggaa
     4141 aagggcaagt ccaagaaact gaagagtgtg aaagagctgc tggggatcac catcatggaa
     4201 agaagcagct tcgagaagaa tcccatcgac tttctggaag ccaagggcta caaagaagtg
     4261 aaaaaggacc tgatcatcaa gctgcctaag tactccctgt tcgagctgga aaacggccgg
     4321 aagagaatgc tggcctctgc cggcgaactg cagaagggaa acgaactggc cctgccctcc
     4381 aaatatgtga acttcctgta cctggccagc cactatgaga agctgaaggg ctcccccgag
     4441 gataatgagc agaaacagct gtttgtggaa cagcacaagc actacctgga cgagatcatc
     4501 gagcagatca gcgagttctc caagagagtg atcctggccg acgctaatct ggacaaagtg
     4561 ctgtccgcct acaacaagca ccgggataag cccatcagag agcaggccga gaatatcatc
     4621 cacctgttta ccctgaccaa tctgggagcc cctgccgcct tcaagtactt tgacaccacc
     4681 atcgaccgga agaggtacac cagcaccaaa gaggtgctgg acgccaccct gatccaccag
     4741 agcatcaccg gcctgtacga gacacggatc gacctgtctc agctgggagg tgacagcggc
     4801 gggagcggcg ggagcggggg gagcactaat ctgagcgaca tcattgagaa ggagactggg
     4861 aaacagctgg tcattcagga gtccatcctg atgctgcctg aggaggtgga ggaagtgatc
     4921 ggcaacaagc cagagtctga catcctggtg cacaccgcct acgacgagtc cacagatgag
     4981 aatgtgatgc tgctgacctc tgacgccccc gagtataagc cttgggccct ggtcatccag
     5041 gattctaacg gcgagaataa gatcaagatg ctgagcggag gatccggagg atctggaggc
     5101 agcaccaacc tgtctgacat catcgagaag gagacaggca agcagctggt catccaggag
     5161 agcatcctga tgctgcccga agaagtcgaa gaagtgatcg gaaacaagcc tgagagcgat
     5221 atcctggtcc ataccgccta cgacgagagt accgacgaaa atgtgatgct gctgacatcc
     5281 gacgccccag agtataagcc ctgggctctg gtcatccagg attccaacgg agagaacaaa
     5341 atcaaaatgc tgtctggcgg ctcaaaaaga accgccgacg gcagcgaatt cgagcccaag
     5401 aagaagagga aagtctaacc ggtcatcatc accatcacca ttgagtttaa acccgctgat
     5461 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     5521 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     5581 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     5641 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     5701 aggcggaaag aaccagctgg ggctcgatac cgtcgacctc tagctagagc ttggcgtaat
     5761 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     5821 gagccggaag cataaagtgt aaagcctagg gtgcctaatg agtgagctaa ctcacattaa
     5881 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5941 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctacaga catgataaga
     6001 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     6061 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttggg
     6121 gtgggcgaag aactccagca tgagatcccc gcgctggagg atcatccagc cggcgtcccg
     6181 gaaaacgatt ccgaagccca acctttcata gaaggcggcg gtggaatcga aatctcgtag
     6241 cacgtgtcag tcctgctcct cggccacgaa gtgcttagcc ctcccacaca taaccagagg
     6301 gcagcaattc acgaatccca actgccgtcg gctgtccatc actgtccttc actatggctt
     6361 tgatcccagg atgcagatcg agaagcacct gtcggcaccg tccgcagggg ctcaagatgc
     6421 ccctgttctc atttccgatc gcgacgatac aagtcaggtt gccagctgcc gcagcagcag
     6481 cagtgcccag caccacgagt tctgcacaag gtcccccagt aaaatgatat acattgacac
     6541 cagtgaagat gcggccgtcg ctagagagag ctgcgctggc gacgctgtag tcttcagaga
     6601 tggggatgct gttgattgta gccgttgctc tttcaatgag ggtggattct tcttgagaca
     6661 aaggcttggc catggtttag ttcctcacct tgtcgtatta tactatgccg atatactatg
     6721 ccgatgatta attgtcaaca cgtgctgatc agatccgaaa atggatatac aagctcccgg
     6781 gagctttttg caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc
     6841 agaggcagag gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg
     6901 agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat
     6961 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
     7021 ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
     7081 ggagcctggg gactttccac accctaactg acacacattc cacagaattc actgactcgc
     7141 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     7201 tatccacaga atcaggggat aacgcaggaa agaacatgta agtgatctcc aaaaaataag
     7261 tactttttga ctgtaaataa aattgtaagg agtaaaaagt actttttttt ctaaaaaaat
     7321 gtaattaagt aaaagtaaaa gtattgattt ttaattgtac tcaagtaaag taaaaatccc
     7381 caaaaataat acttaagtac agtaatcaag taaaattact caagtacttt acacctctgc
     7441 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     7501 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     7561 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     7621 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     7681 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     7741 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     7801 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     7861 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     7921 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     7981 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     8041 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     8101 atcttttcta cggggtctga cactcagtgg aacgaaaact cacgttaagg gattttggtc
     8161 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     8221 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     8281 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     8341 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcgg
     8401 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     8461 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     8521 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     8581 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     8641 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     8701 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     8761 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     8821 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     8881 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     8941 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     9001 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     9061 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     9121 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     9181 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     9241 gtgccacctg acgtcgacgg atcgggagat cgatctcccg atcccctagg gtcgactctc
     9301 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
     9361 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
     9421 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
     9481 agatatacgc gggtaccggc atatggttct tgacagaggt gtaaaaagta ctcaaaaatt
     9541 ttactcaagt gaaagtacaa gtacttaggg aaaattttac tcaattaaaa gtaaaagtat
     9601 ctggctagaa tcttacttga gtaaaagtaa aaaagtactc cattaaaatt gtacttgagt
     9661 attaacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
     9721 agtttatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     9781 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     9841 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     9901 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     9961 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
    10021 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
    10081 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
    10141 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
    10201 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt
    10261 agtgaaccgt cagatccgct agagatccg
//
