LOCUS       sequence-217432-        9409 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses hA3A-eBE-Y130F in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9409)
  AUTHORS   Wang X, Li J, Wang Y, Yang B, Wei J, Wu J, Wang R, Huang X, Chen J, 
            Yang L
  TITLE     Efficient base editing in methylated regions with a human 
            APOBEC3A-Cas9 fusion.
  JOURNAL   Nat Biotechnol. 2018 Aug 20. pii: nbt.4198. doi: 10.1038/nbt.4198.
  PUBMED    30125268
REFERENCE   2  (bases 1 to 9409)
  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. 2018 Aug 20. pii: nbt.4198. doi: 10.1038/nbt.4198."
COMMENT     Sequence Label: pCMV-hA3A-eBE-Y130F
FEATURES             Location/Qualifiers
     source          1..9409
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     146..166
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     237..256
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        237..255
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      263..272
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             269..865
                     /codon_start=1
                     /gene="human APOBEC3A"
                     /gene_synonym="A3A"
                     /product="human cytidine deaminase (C to U editing enzyme) 
                     that efficiently edits methylated cytosines (Wang et al., 
                     2018)"
                     /label=APOBEC3A
                     /note="also known as hA3A"
                     /translation="MEASPASGPRHLMDPHIFTSNFNNGIGRHKTYLCYEVERLDNGTS
                     VKMDQHRGFLHNQAKNLLCGFYGRHAELRFLDLVPSLQLDPAQIYRVTWFISWSPCFSW
                     GCAGEVRAFLQENTHVRLRIFAARIFDYDPLYKEALQMLRDAGAQVSIMTYDEFKHCWD
                     TFVDHQGCPFQPWDGLDEHSQALSGRLRAILQNQGN"
     CDS             join(914..4312,4313..4315,4316..4912,4913,4914..4916,
                     4917..4918,4919..4921,4922..5014)
                     /codon_start=1
                     /product="Streptococcus pyogenes Cas9 endonuclease with the
                     D1135V/R1335Q/T1337R mutations"
                     /label=Cas9 VQR
                     /note="has an altered recognition specificity for the 
                     protospacer adjacent motif (PAM) (Kleinstiver et al., 
                     2015)"
                     /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             join(914..937,938..940,941..5014)
                     /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             5027..5275
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5288..5308
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5315..5368
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             5369..5617
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5618..5638
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5639..5695
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             5696..5944
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5945..5965
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5966..6019
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             6020..6268
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6281..6301
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6324..6341)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6330..6554
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6625..6641)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6625..6641)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6638..6660)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6649..6665
                     /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(6673..6679,6680..6697,6698..6703))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6718..6739
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6856..6873)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7027..7615)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7107..7126)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7786..8577,8578..8646))
                     /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     8409..8428
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8647..8751)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8830..8849)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        9021..9400
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(9401..9409,1..195)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
       61 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
      121 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
      181 tatataagca gagctggttt agtgaaccgt cagatccgct agagatccgc ggccgctaat
      241 acgactcact atagggagag ccgccaccat ggaagccagc ccagcatccg ggcccagaca
      301 cttgatggat ccacacatat tcacttccaa ctttaacaat ggcattggaa ggcataagac
      361 ctacctgtgc tacgaagtgg agcgcctgga caatggcacc tcggtcaaga tggaccagca
      421 caggggcttt ctacacaacc aggctaagaa tcttctctgt ggcttttacg gccgccatgc
      481 ggagctgcgc ttcttggacc tggttccttc tttgcagttg gacccggccc agatctacag
      541 ggtcacttgg ttcatctcct ggagcccctg cttctcctgg ggctgtgccg gggaagtgcg
      601 tgcgttcctt caggagaaca cacacgtgag actgcgtatc ttcgctgccc gcatctttga
      661 ttacgacccc ctatataagg aggcactgca aatgctgcgg gatgctgggg cccaagtctc
      721 catcatgacc tacgatgaat ttaagcactg ctgggacacc tttgtggacc accagggatg
      781 tcccttccag ccctgggatg gactagatga gcacagccaa gccctgagtg ggaggctgcg
      841 ggccattctc cagaatcagg gaaacagcgg cagcgagact cccgggacct cagagtccgc
      901 cacacccgaa agtgataaaa agtattctat tggtttagcc atcggcacta attccgttgg
      961 atgggctgtc ataaccgatg aatacaaagt accttcaaag aaatttaagg tgttggggaa
     1021 cacagaccgt cattcgatta aaaagaatct tatcggtgcc ctcctattcg atagtggcga
     1081 aacggcagag gcgactcgcc tgaaacgaac cgctcggaga aggtatacac gtcgcaagaa
     1141 ccgaatatgt tacttacaag aaatttttag caatgagatg gccaaagttg acgattcttt
     1201 ctttcaccgt ttggaagagt ccttccttgt cgaagaggac aagaaacatg aacggcaccc
     1261 catctttgga aacatagtag atgaggtggc atatcatgaa aagtacccaa cgatttatca
     1321 cctcagaaaa aagctagttg actcaactga taaagcggac ctgaggttaa tctacttggc
     1381 tcttgcccat atgataaagt tccgtgggca ctttctcatt gagggtgatc taaatccgga
     1441 caactcggat gtcgacaaac tgttcatcca gttagtacaa acctataatc agttgtttga
     1501 agagaaccct ataaatgcaa gtggcgtgga tgcgaaggct attcttagcg cccgcctctc
     1561 taaatcccga cggctagaaa acctgatcgc acaattaccc ggagagaaga aaaatgggtt
     1621 gttcggtaac cttatagcgc tctcactagg cctgacacca aattttaagt cgaacttcga
     1681 cttagctgaa gatgccaaat tgcagcttag taaggacacg tacgatgacg atctcgacaa
     1741 tctactggca caaattggag atcagtatgc ggacttattt ttggctgcca aaaaccttag
     1801 cgatgcaatc ctcctatctg acatactgag agttaatact gagattacca aggcgccgtt
     1861 atccgcttca atgatcaaaa ggtacgatga acatcaccaa gacttgacac ttctcaaggc
     1921 cctagtccgt cagcaactgc ctgagaaata taaggaaata ttctttgatc agtcgaaaaa
     1981 cgggtacgca ggttatattg acggcggagc gagtcaagag gaattctaca agtttatcaa
     2041 acccatatta gagaagatgg atgggacgga agagttgctt gtaaaactca atcgcgaaga
     2101 tctactgcga aagcagcgga ctttcgacaa cggtagcatt ccacatcaaa tccacttagg
     2161 cgaattgcat gctatactta gaaggcagga ggatttttat ccgttcctca aagacaatcg
     2221 tgaaaagatt gagaaaatcc taacctttcg cataccttac tatgtgggac ccctggcccg
     2281 agggaactct cggttcgcat ggatgacaag aaagtccgaa gaaacgatta ctccatggaa
     2341 ttttgaggaa gttgtcgata aaggtgcgtc agctcaatcg ttcatcgaga ggatgaccaa
     2401 ctttgacaag aatttaccga acgaaaaagt attgcctaag cacagtttac tttacgagta
     2461 tttcacagtg tacaatgaac tcacgaaagt taagtatgtc actgagggca tgcgtaaacc
     2521 cgcctttcta agcggagaac agaagaaagc aatagtagat ctgttattca agaccaaccg
     2581 caaagtgaca gttaagcaat tgaaagagga ctactttaag aaaattgaat gcttcgattc
     2641 tgtcgagatc tccggggtag aagatcgatt taatgcgtca cttggtacgt atcatgacct
     2701 cctaaagata attaaagata aggacttcct ggataacgaa gagaatgaag atatcttaga
     2761 agatatagtg ttgactctta ccctctttga agatcgggaa atgattgagg aaagactaaa
     2821 aacatacgct cacctgttcg acgataaggt tatgaaacag ttaaagaggc gtcgctatac
     2881 gggctgggga cgattgtcgc ggaaacttat caacgggata agagacaagc aaagtggtaa
     2941 aactattctc gattttctaa agagcgacgg cttcgccaat aggaacttta tgcagctgat
     3001 ccatgatgac tctttaacct tcaaagagga tatacaaaag gcacaggttt ccggacaagg
     3061 ggactcattg cacgaacata ttgcgaatct tgctggttcg ccagccatca aaaagggcat
     3121 actccagaca gtcaaagtag tggatgagct agttaaggtc atgggacgtc acaaaccgga
     3181 aaacattgta atcgagatgg cacgcgaaaa tcaaacgact cagaaggggc aaaaaaacag
     3241 tcgagagcgg atgaagagaa tagaagaggg tattaaagaa ctgggcagcc agatcttaaa
     3301 ggagcatcct gtggaaaata cccaattgca gaacgagaaa ctttacctct attacctaca
     3361 aaatggaagg gacatgtatg ttgatcagga actggacata aaccgtttat ctgattacga
     3421 cgtcgatcac attgtacccc aatccttttt gaaggacgat tcaatcgaca ataaagtgct
     3481 tacacgctcg gataagaacc gagggaaaag tgacaatgtt ccaagcgagg aagtcgtaaa
     3541 gaaaatgaag aactattggc ggcagctcct aaatgcgaaa ctgataacgc aaagaaagtt
     3601 cgataactta actaaagctg agaggggtgg cttgtctgaa cttgacaagg ccggatttat
     3661 taaacgtcag ctcgtggaaa cccgccaaat cacaaagcat gttgcacaga tactagattc
     3721 ccgaatgaat acgaaatacg acgagaacga taagctgatt cgggaagtca aagtaatcac
     3781 tttaaagtca aaattggtgt cggacttcag aaaggatttt caattctata aagttaggga
     3841 gataaataac taccaccatg cgcacgacgc ttatcttaat gccgtcgtag ggaccgcact
     3901 cattaagaaa tacccgaagc tagaaagtga gtttgtgtat ggtgattaca aagtttatga
     3961 cgtccgtaag atgatcgcga aaagcgaaca ggagataggc aaggctacag ccaaatactt
     4021 cttttattct aacattatga atttctttaa gacggaaatc actctggcaa acggagagat
     4081 acgcaaacga cctttaattg aaaccaatgg ggagacaggt gaaatcgtat gggataaggg
     4141 ccgggacttc gcgacggtga gaaaagtttt gtccatgccc caagtcaaca tagtaaagaa
     4201 aactgaggtg cagaccggag ggttttcaaa ggaatcgatt cttccaaaaa ggaatagtga
     4261 taagctcatc gctcgtaaaa aggactggga cccgaaaaag tacggtggct tcgatagccc
     4321 tacagttgcc tattctgtcc tagtagtggc aaaagttgag aagggaaaat ccaagaaact
     4381 gaagtcagtc aaagaattat tggggataac gattatggag cgctcgtctt ttgaaaagaa
     4441 ccccatcgac ttccttgagg cgaaaggtta caaggaagta aaaaaggatc tcataattaa
     4501 actaccaaag tatagtctgt ttgagttaga aaatggccga aaacggatgt tggctagcgc
     4561 cggagagctt caaaagggga acgaactcgc actaccgtct aaatacgtga atttcctgta
     4621 tttagcgtcc cattacgaga agttgaaagg ttcacctgaa gataacgaac agaagcaact
     4681 ttttgttgag cagcacaaac attatctcga cgaaatcata gagcaaattt cggaattcag
     4741 taagagagtc atcctagctg atgccaatct ggacaaagta ttaagcgcat acaacaagca
     4801 cagggataaa cccatacgtg agcaggcgga aaatattatc catttgttta ctcttaccaa
     4861 cctcggcgct ccagccgcat tcaagtattt tgacacaacg atagatcgca aacgatacac
     4921 ttctaccaag gaggtgctag acgcgacact gattcaccaa tccatcacgg gattatatga
     4981 aactcggata gatttgtcac agcttggggg tgactctggt ggttctacta atctgtcaga
     5041 tattattgaa aaggagaccg gtaagcaact ggttatccag gaatccatcc tcatgctccc
     5101 agaggaggtg gaagaagtca ttgggaacaa gccggaaagc gatatactcg tgcacaccgc
     5161 ctacgacgag agcaccgacg agaatgtcat gcttctgact agcgacgccc ctgaatacaa
     5221 gccttgggct ctggtcatac aggatagcaa cggtgagaac aagattaaga tgctctctgg
     5281 tggttctccc aagaagaaga ggaaagtcgg gcccgagggc agaggaagtc tgctaacatg
     5341 cggtgacgtc gaggagaatc ctggcccaac taatctgtca gatattattg aaaaggagac
     5401 cggtaagcaa ctggttatcc aggaatccat cctcatgctc ccagaggagg tggaagaagt
     5461 cattgggaac aagccggaaa gcgatatact cgtgcacacc gcctacgacg agagcaccga
     5521 cgagaatgtc atgcttctga ctagcgacgc ccctgaatac aagccttggg ctctggtcat
     5581 acaggatagc aacggtgaga acaagattaa gatgctcccc aagaagaaga ggaaagtcgc
     5641 tactaacttc agcctgctga agcaggctgg agacgtggag gagaaccctg gacctaccaa
     5701 cctgtccgat atcattgaga aagagaccgg caaacagctg gtgatccagg agagcatcct
     5761 gatgctgccc gaagaggtgg aggaagtgat cggcaacaag cccgagtccg acatcctggt
     5821 gcacacagcc tatgatgaat ccaccgacga gaacgtgatg ctgctgacct ccgatgctcc
     5881 cgagtataaa ccctgggcac tggtgatcca ggactctaat ggagagaaca agatcaagat
     5941 gctgcccaag aagaagagga aagtcgaagg acggggctcc ctcctgacct gtggcgatgt
     6001 ggaagagaac cccggcccca caaacctcag tgacattatc gagaaggaaa caggaaaaca
     6061 gctcgtcatt caagaatcta ttcttatgtt gcctgaggaa gtcgaagagg ttattggcaa
     6121 taaacctgaa tctgatattc ttgtccatac cgcttacgat gagtccacag atgaaaatgt
     6181 tatgctgctc acatctgacg caccagagta caaaccatgg gcgctcgtta ttcaagattc
     6241 caacggcgaa aacaaaatca aaatgctttc tggtggttct cctaaaaaaa aaagaaaggt
     6301 gtgagtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     6361 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     6421 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     6481 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     6541 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctcgatac cgtcgacctc
     6601 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     6661 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctagg gtgcctaatg
     6721 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     6781 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     6841 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6901 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6961 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     7021 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     7081 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     7141 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     7201 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     7261 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     7321 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     7381 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     7441 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     7501 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     7561 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     7621 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7681 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     7741 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7801 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7861 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7921 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7981 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     8041 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     8101 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     8161 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     8221 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     8281 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     8341 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     8401 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     8461 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     8521 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     8581 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     8641 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     8701 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     8761 ccgaaaagtg ccacctgacg tcgacggatc gggagatcga tctcccgatc ccctagggtc
     8821 gactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg
     8881 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt
     8941 gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt
     9001 acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac
     9061 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     9121 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     9181 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     9241 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     9301 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     9361 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcgg
//
