LOCUS       sequence-217446-        9409 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses hA3A-eBE-Y132D 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-Y132D
FEATURES             Location/Qualifiers
     source          1..9409
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        123..502
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        503..706
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     657..677
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     748..767
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        748..766
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      774..783
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             780..1376
                     /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
                     GCAGEVRAFLQENTHVRLRIFAARIYDDDPLYKEALQMLRDAGAQVSIMTYDEFKHCWD
                     TFVDHQGCPFQPWDGLDEHSQALSGRLRAILQNQGN"
     CDS             join(1425..4823,4824..4826,4827..5423,5424,5425..5427,
                     5428..5429,5430..5432,5433..5525)
                     /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(1425..1448,1449..1451,1452..5525)
                     /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             5538..5786
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5799..5819
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5826..5879
                     /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             5880..6128
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6129..6149
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6150..6206
                     /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             6207..6455
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6456..6476
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6477..6530
                     /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             6531..6779
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6792..6812
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6835..6852)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6841..7065
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7136..7152)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7136..7152)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7149..7171)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7160..7176
                     /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(7184..7190,7191..7208,7209..7214))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7229..7250
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7367..7384)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7538..8126)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7618..7637)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8297..9088,9089..9157))
                     /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     8920..8939
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9158..9262)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9341..9360)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
       61 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg
      121 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
      181 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      241 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      301 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      361 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      421 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      481 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
      541 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
      601 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
      661 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg
      721 tcagatccgc tagagatccg cggccgctaa tacgactcac tatagggaga gccgccacca
      781 tggaagccag cccagcatcc gggcccagac acttgatgga tccacacata ttcacttcca
      841 actttaacaa tggcattgga aggcataaga cctacctgtg ctacgaagtg gagcgcctgg
      901 acaatggcac ctcggtcaag atggaccagc acaggggctt tctacacaac caggctaaga
      961 atcttctctg tggcttttac ggccgccatg cggagctgcg cttcttggac ctggttcctt
     1021 ctttgcagtt ggacccggcc cagatctaca gggtcacttg gttcatctcc tggagcccct
     1081 gcttctcctg gggctgtgcc ggggaagtgc gtgcgttcct tcaggagaac acacacgtga
     1141 gactgcgtat cttcgctgcc cgcatctatg atgatgaccc cctatataag gaggcactgc
     1201 aaatgctgcg ggatgctggg gcccaagtct ccatcatgac ctacgatgaa tttaagcact
     1261 gctgggacac ctttgtggac caccagggat gtcccttcca gccctgggat ggactagatg
     1321 agcacagcca agccctgagt gggaggctgc gggccattct ccagaatcag ggaaacagcg
     1381 gcagcgagac tcccgggacc tcagagtccg ccacacccga aagtgataaa aagtattcta
     1441 ttggtttagc catcggcact aattccgttg gatgggctgt cataaccgat gaatacaaag
     1501 taccttcaaa gaaatttaag gtgttgggga acacagaccg tcattcgatt aaaaagaatc
     1561 ttatcggtgc cctcctattc gatagtggcg aaacggcaga ggcgactcgc ctgaaacgaa
     1621 ccgctcggag aaggtataca cgtcgcaaga accgaatatg ttacttacaa gaaattttta
     1681 gcaatgagat ggccaaagtt gacgattctt tctttcaccg tttggaagag tccttccttg
     1741 tcgaagagga caagaaacat gaacggcacc ccatctttgg aaacatagta gatgaggtgg
     1801 catatcatga aaagtaccca acgatttatc acctcagaaa aaagctagtt gactcaactg
     1861 ataaagcgga cctgaggtta atctacttgg ctcttgccca tatgataaag ttccgtgggc
     1921 actttctcat tgagggtgat ctaaatccgg acaactcgga tgtcgacaaa ctgttcatcc
     1981 agttagtaca aacctataat cagttgtttg aagagaaccc tataaatgca agtggcgtgg
     2041 atgcgaaggc tattcttagc gcccgcctct ctaaatcccg acggctagaa aacctgatcg
     2101 cacaattacc cggagagaag aaaaatgggt tgttcggtaa ccttatagcg ctctcactag
     2161 gcctgacacc aaattttaag tcgaacttcg acttagctga agatgccaaa ttgcagctta
     2221 gtaaggacac gtacgatgac gatctcgaca atctactggc acaaattgga gatcagtatg
     2281 cggacttatt tttggctgcc aaaaacctta gcgatgcaat cctcctatct gacatactga
     2341 gagttaatac tgagattacc aaggcgccgt tatccgcttc aatgatcaaa aggtacgatg
     2401 aacatcacca agacttgaca cttctcaagg ccctagtccg tcagcaactg cctgagaaat
     2461 ataaggaaat attctttgat cagtcgaaaa acgggtacgc aggttatatt gacggcggag
     2521 cgagtcaaga ggaattctac aagtttatca aacccatatt agagaagatg gatgggacgg
     2581 aagagttgct tgtaaaactc aatcgcgaag atctactgcg aaagcagcgg actttcgaca
     2641 acggtagcat tccacatcaa atccacttag gcgaattgca tgctatactt agaaggcagg
     2701 aggattttta tccgttcctc aaagacaatc gtgaaaagat tgagaaaatc ctaacctttc
     2761 gcatacctta ctatgtggga cccctggccc gagggaactc tcggttcgca tggatgacaa
     2821 gaaagtccga agaaacgatt actccatgga attttgagga agttgtcgat aaaggtgcgt
     2881 cagctcaatc gttcatcgag aggatgacca actttgacaa gaatttaccg aacgaaaaag
     2941 tattgcctaa gcacagttta ctttacgagt atttcacagt gtacaatgaa ctcacgaaag
     3001 ttaagtatgt cactgagggc atgcgtaaac ccgcctttct aagcggagaa cagaagaaag
     3061 caatagtaga tctgttattc aagaccaacc gcaaagtgac agttaagcaa ttgaaagagg
     3121 actactttaa gaaaattgaa tgcttcgatt ctgtcgagat ctccggggta gaagatcgat
     3181 ttaatgcgtc acttggtacg tatcatgacc tcctaaagat aattaaagat aaggacttcc
     3241 tggataacga agagaatgaa gatatcttag aagatatagt gttgactctt accctctttg
     3301 aagatcggga aatgattgag gaaagactaa aaacatacgc tcacctgttc gacgataagg
     3361 ttatgaaaca gttaaagagg cgtcgctata cgggctgggg acgattgtcg cggaaactta
     3421 tcaacgggat aagagacaag caaagtggta aaactattct cgattttcta aagagcgacg
     3481 gcttcgccaa taggaacttt atgcagctga tccatgatga ctctttaacc ttcaaagagg
     3541 atatacaaaa ggcacaggtt tccggacaag gggactcatt gcacgaacat attgcgaatc
     3601 ttgctggttc gccagccatc aaaaagggca tactccagac agtcaaagta gtggatgagc
     3661 tagttaaggt catgggacgt cacaaaccgg aaaacattgt aatcgagatg gcacgcgaaa
     3721 atcaaacgac tcagaagggg caaaaaaaca gtcgagagcg gatgaagaga atagaagagg
     3781 gtattaaaga actgggcagc cagatcttaa aggagcatcc tgtggaaaat acccaattgc
     3841 agaacgagaa actttacctc tattacctac aaaatggaag ggacatgtat gttgatcagg
     3901 aactggacat aaaccgttta tctgattacg acgtcgatca cattgtaccc caatcctttt
     3961 tgaaggacga ttcaatcgac aataaagtgc ttacacgctc ggataagaac cgagggaaaa
     4021 gtgacaatgt tccaagcgag gaagtcgtaa agaaaatgaa gaactattgg cggcagctcc
     4081 taaatgcgaa actgataacg caaagaaagt tcgataactt aactaaagct gagaggggtg
     4141 gcttgtctga acttgacaag gccggattta ttaaacgtca gctcgtggaa acccgccaaa
     4201 tcacaaagca tgttgcacag atactagatt cccgaatgaa tacgaaatac gacgagaacg
     4261 ataagctgat tcgggaagtc aaagtaatca ctttaaagtc aaaattggtg tcggacttca
     4321 gaaaggattt tcaattctat aaagttaggg agataaataa ctaccaccat gcgcacgacg
     4381 cttatcttaa tgccgtcgta gggaccgcac tcattaagaa atacccgaag ctagaaagtg
     4441 agtttgtgta tggtgattac aaagtttatg acgtccgtaa gatgatcgcg aaaagcgaac
     4501 aggagatagg caaggctaca gccaaatact tcttttattc taacattatg aatttcttta
     4561 agacggaaat cactctggca aacggagaga tacgcaaacg acctttaatt gaaaccaatg
     4621 gggagacagg tgaaatcgta tgggataagg gccgggactt cgcgacggtg agaaaagttt
     4681 tgtccatgcc ccaagtcaac atagtaaaga aaactgaggt gcagaccgga gggttttcaa
     4741 aggaatcgat tcttccaaaa aggaatagtg ataagctcat cgctcgtaaa aaggactggg
     4801 acccgaaaaa gtacggtggc ttcgatagcc ctacagttgc ctattctgtc ctagtagtgg
     4861 caaaagttga gaagggaaaa tccaagaaac tgaagtcagt caaagaatta ttggggataa
     4921 cgattatgga gcgctcgtct tttgaaaaga accccatcga cttccttgag gcgaaaggtt
     4981 acaaggaagt aaaaaaggat ctcataatta aactaccaaa gtatagtctg tttgagttag
     5041 aaaatggccg aaaacggatg ttggctagcg ccggagagct tcaaaagggg aacgaactcg
     5101 cactaccgtc taaatacgtg aatttcctgt atttagcgtc ccattacgag aagttgaaag
     5161 gttcacctga agataacgaa cagaagcaac tttttgttga gcagcacaaa cattatctcg
     5221 acgaaatcat agagcaaatt tcggaattca gtaagagagt catcctagct gatgccaatc
     5281 tggacaaagt attaagcgca tacaacaagc acagggataa acccatacgt gagcaggcgg
     5341 aaaatattat ccatttgttt actcttacca acctcggcgc tccagccgca ttcaagtatt
     5401 ttgacacaac gatagatcgc aaacgataca cttctaccaa ggaggtgcta gacgcgacac
     5461 tgattcacca atccatcacg ggattatatg aaactcggat agatttgtca cagcttgggg
     5521 gtgactctgg tggttctact aatctgtcag atattattga aaaggagacc ggtaagcaac
     5581 tggttatcca ggaatccatc ctcatgctcc cagaggaggt ggaagaagtc attgggaaca
     5641 agccggaaag cgatatactc gtgcacaccg cctacgacga gagcaccgac gagaatgtca
     5701 tgcttctgac tagcgacgcc cctgaataca agccttgggc tctggtcata caggatagca
     5761 acggtgagaa caagattaag atgctctctg gtggttctcc caagaagaag aggaaagtcg
     5821 ggcccgaggg cagaggaagt ctgctaacat gcggtgacgt cgaggagaat cctggcccaa
     5881 ctaatctgtc agatattatt gaaaaggaga ccggtaagca actggttatc caggaatcca
     5941 tcctcatgct cccagaggag gtggaagaag tcattgggaa caagccggaa agcgatatac
     6001 tcgtgcacac cgcctacgac gagagcaccg acgagaatgt catgcttctg actagcgacg
     6061 cccctgaata caagccttgg gctctggtca tacaggatag caacggtgag aacaagatta
     6121 agatgctccc caagaagaag aggaaagtcg ctactaactt cagcctgctg aagcaggctg
     6181 gagacgtgga ggagaaccct ggacctacca acctgtccga tatcattgag aaagagaccg
     6241 gcaaacagct ggtgatccag gagagcatcc tgatgctgcc cgaagaggtg gaggaagtga
     6301 tcggcaacaa gcccgagtcc gacatcctgg tgcacacagc ctatgatgaa tccaccgacg
     6361 agaacgtgat gctgctgacc tccgatgctc ccgagtataa accctgggca ctggtgatcc
     6421 aggactctaa tggagagaac aagatcaaga tgctgcccaa gaagaagagg aaagtcgaag
     6481 gacggggctc cctcctgacc tgtggcgatg tggaagagaa ccccggcccc acaaacctca
     6541 gtgacattat cgagaaggaa acaggaaaac agctcgtcat tcaagaatct attcttatgt
     6601 tgcctgagga agtcgaagag gttattggca ataaacctga atctgatatt cttgtccata
     6661 ccgcttacga tgagtccaca gatgaaaatg ttatgctgct cacatctgac gcaccagagt
     6721 acaaaccatg ggcgctcgtt attcaagatt ccaacggcga aaacaaaatc aaaatgcttt
     6781 ctggtggttc tcctaaaaaa aaaagaaagg tgtgagttta aacccgctga tcagcctcga
     6841 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     6901 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     6961 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     7021 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     7081 gaaccagctg gggctcgata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     7141 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     7201 gcataaagtg taaagcctag ggtgcctaat gagtgagcta actcacatta attgcgttgc
     7261 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     7321 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     7381 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     7441 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     7501 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     7561 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     7621 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     7681 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     7741 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     7801 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     7861 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     7921 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     7981 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     8041 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     8101 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     8161 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     8221 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     8281 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     8341 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     8401 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     8461 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     8521 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     8581 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     8641 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     8701 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     8761 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     8821 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     8881 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     8941 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     9001 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     9061 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     9121 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     9181 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     9241 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     9301 cgggagatcg atctcccgat cccctagggt cgactctcag tacaatctgc tctgatgccg
     9361 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctga
//
