LOCUS       sequence-217434-        8442 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses hA3A-BE3-C101S in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8442)
  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 8442)
  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-BE3-C101S
FEATURES             Location/Qualifiers
     source          1..8442
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        142..521
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        522..725
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     676..696
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     767..786
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        767..785
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      793..802
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             799..1395
                     /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
                     VKMDQHRGFLHNQAKNLLCGFYGRHAELRFLDLVPSLQLDPAQIYRVTWFISWSPSFSW
                     GCAGEVRAFLQENTHVRLRIFAARIYDYDPLYKEALQMLRDAGAQVSIMTYDEFKHCWD
                     TFVDHQGCPFQPWDGLDEHSQALSGRLRAILQNQGN"
     CDS             join(1444..4842,4843..4845,4846..5442,5443,5444..5446,
                     5447..5448,5449..5451,5452..5544)
                     /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(1444..1467,1468..1470,1471..5544)
                     /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             5557..5805
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5818..5838
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5847..5864
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5887..5904)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5893..6117
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6188..6204)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6188..6204)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6201..6223)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6212..6228
                     /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(6236..6242,6243..6260,6261..6266))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6281..6302
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6419..6436)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6590..7178)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6670..6689)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7349..8140,8141..8209))
                     /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     7972..7991
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8210..8314)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8393..8412)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
       61 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      121 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      181 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      241 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      301 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      361 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      421 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      481 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      541 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      601 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      661 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      721 gagctggttt agtgaaccgt cagatccgct agagatccgc ggccgctaat acgactcact
      781 atagggagag ccgccaccat ggaagccagc ccagcatccg ggcccagaca cttgatggat
      841 ccacacatat tcacttccaa ctttaacaat ggcattggaa ggcataagac ctacctgtgc
      901 tacgaagtgg agcgcctgga caatggcacc tcggtcaaga tggaccagca caggggcttt
      961 ctacacaacc aggctaagaa tcttctctgt ggcttttacg gccgccatgc ggagctgcgc
     1021 ttcttggacc tggttccttc tttgcagttg gacccggccc agatctacag ggtcacttgg
     1081 ttcatctcct ggagccccag cttctcctgg ggctgtgccg gggaagtgcg tgcgttcctt
     1141 caggagaaca cacacgtgag actgcgtatc ttcgctgccc gcatctatga ttacgacccc
     1201 ctatataagg aggcactgca aatgctgcgg gatgctgggg cccaagtctc catcatgacc
     1261 tacgatgaat ttaagcactg ctgggacacc tttgtggacc accagggatg tcccttccag
     1321 ccctgggatg gactagatga gcacagccaa gccctgagtg ggaggctgcg ggccattctc
     1381 cagaatcagg gaaacagcgg cagcgagact cccgggacct cagagtccgc cacacccgaa
     1441 agtgataaaa agtattctat tggtttagcc atcggcacta attccgttgg atgggctgtc
     1501 ataaccgatg aatacaaagt accttcaaag aaatttaagg tgttggggaa cacagaccgt
     1561 cattcgatta aaaagaatct tatcggtgcc ctcctattcg atagtggcga aacggcagag
     1621 gcgactcgcc tgaaacgaac cgctcggaga aggtatacac gtcgcaagaa ccgaatatgt
     1681 tacttacaag aaatttttag caatgagatg gccaaagttg acgattcttt ctttcaccgt
     1741 ttggaagagt ccttccttgt cgaagaggac aagaaacatg aacggcaccc catctttgga
     1801 aacatagtag atgaggtggc atatcatgaa aagtacccaa cgatttatca cctcagaaaa
     1861 aagctagttg actcaactga taaagcggac ctgaggttaa tctacttggc tcttgcccat
     1921 atgataaagt tccgtgggca ctttctcatt gagggtgatc taaatccgga caactcggat
     1981 gtcgacaaac tgttcatcca gttagtacaa acctataatc agttgtttga agagaaccct
     2041 ataaatgcaa gtggcgtgga tgcgaaggct attcttagcg cccgcctctc taaatcccga
     2101 cggctagaaa acctgatcgc acaattaccc ggagagaaga aaaatgggtt gttcggtaac
     2161 cttatagcgc tctcactagg cctgacacca aattttaagt cgaacttcga cttagctgaa
     2221 gatgccaaat tgcagcttag taaggacacg tacgatgacg atctcgacaa tctactggca
     2281 caaattggag atcagtatgc ggacttattt ttggctgcca aaaaccttag cgatgcaatc
     2341 ctcctatctg acatactgag agttaatact gagattacca aggcgccgtt atccgcttca
     2401 atgatcaaaa ggtacgatga acatcaccaa gacttgacac ttctcaaggc cctagtccgt
     2461 cagcaactgc ctgagaaata taaggaaata ttctttgatc agtcgaaaaa cgggtacgca
     2521 ggttatattg acggcggagc gagtcaagag gaattctaca agtttatcaa acccatatta
     2581 gagaagatgg atgggacgga agagttgctt gtaaaactca atcgcgaaga tctactgcga
     2641 aagcagcgga ctttcgacaa cggtagcatt ccacatcaaa tccacttagg cgaattgcat
     2701 gctatactta gaaggcagga ggatttttat ccgttcctca aagacaatcg tgaaaagatt
     2761 gagaaaatcc taacctttcg cataccttac tatgtgggac ccctggcccg agggaactct
     2821 cggttcgcat ggatgacaag aaagtccgaa gaaacgatta ctccatggaa ttttgaggaa
     2881 gttgtcgata aaggtgcgtc agctcaatcg ttcatcgaga ggatgaccaa ctttgacaag
     2941 aatttaccga acgaaaaagt attgcctaag cacagtttac tttacgagta tttcacagtg
     3001 tacaatgaac tcacgaaagt taagtatgtc actgagggca tgcgtaaacc cgcctttcta
     3061 agcggagaac agaagaaagc aatagtagat ctgttattca agaccaaccg caaagtgaca
     3121 gttaagcaat tgaaagagga ctactttaag aaaattgaat gcttcgattc tgtcgagatc
     3181 tccggggtag aagatcgatt taatgcgtca cttggtacgt atcatgacct cctaaagata
     3241 attaaagata aggacttcct ggataacgaa gagaatgaag atatcttaga agatatagtg
     3301 ttgactctta ccctctttga agatcgggaa atgattgagg aaagactaaa aacatacgct
     3361 cacctgttcg acgataaggt tatgaaacag ttaaagaggc gtcgctatac gggctgggga
     3421 cgattgtcgc ggaaacttat caacgggata agagacaagc aaagtggtaa aactattctc
     3481 gattttctaa agagcgacgg cttcgccaat aggaacttta tgcagctgat ccatgatgac
     3541 tctttaacct tcaaagagga tatacaaaag gcacaggttt ccggacaagg ggactcattg
     3601 cacgaacata ttgcgaatct tgctggttcg ccagccatca aaaagggcat actccagaca
     3661 gtcaaagtag tggatgagct agttaaggtc atgggacgtc acaaaccgga aaacattgta
     3721 atcgagatgg cacgcgaaaa tcaaacgact cagaaggggc aaaaaaacag tcgagagcgg
     3781 atgaagagaa tagaagaggg tattaaagaa ctgggcagcc agatcttaaa ggagcatcct
     3841 gtggaaaata cccaattgca gaacgagaaa ctttacctct attacctaca aaatggaagg
     3901 gacatgtatg ttgatcagga actggacata aaccgtttat ctgattacga cgtcgatcac
     3961 attgtacccc aatccttttt gaaggacgat tcaatcgaca ataaagtgct tacacgctcg
     4021 gataagaacc gagggaaaag tgacaatgtt ccaagcgagg aagtcgtaaa gaaaatgaag
     4081 aactattggc ggcagctcct aaatgcgaaa ctgataacgc aaagaaagtt cgataactta
     4141 actaaagctg agaggggtgg cttgtctgaa cttgacaagg ccggatttat taaacgtcag
     4201 ctcgtggaaa cccgccaaat cacaaagcat gttgcacaga tactagattc ccgaatgaat
     4261 acgaaatacg acgagaacga taagctgatt cgggaagtca aagtaatcac tttaaagtca
     4321 aaattggtgt cggacttcag aaaggatttt caattctata aagttaggga gataaataac
     4381 taccaccatg cgcacgacgc ttatcttaat gccgtcgtag ggaccgcact cattaagaaa
     4441 tacccgaagc tagaaagtga gtttgtgtat ggtgattaca aagtttatga cgtccgtaag
     4501 atgatcgcga aaagcgaaca ggagataggc aaggctacag ccaaatactt cttttattct
     4561 aacattatga atttctttaa gacggaaatc actctggcaa acggagagat acgcaaacga
     4621 cctttaattg aaaccaatgg ggagacaggt gaaatcgtat gggataaggg ccgggacttc
     4681 gcgacggtga gaaaagtttt gtccatgccc caagtcaaca tagtaaagaa aactgaggtg
     4741 cagaccggag ggttttcaaa ggaatcgatt cttccaaaaa ggaatagtga taagctcatc
     4801 gctcgtaaaa aggactggga cccgaaaaag tacggtggct tcgatagccc tacagttgcc
     4861 tattctgtcc tagtagtggc aaaagttgag aagggaaaat ccaagaaact gaagtcagtc
     4921 aaagaattat tggggataac gattatggag cgctcgtctt ttgaaaagaa ccccatcgac
     4981 ttccttgagg cgaaaggtta caaggaagta aaaaaggatc tcataattaa actaccaaag
     5041 tatagtctgt ttgagttaga aaatggccga aaacggatgt tggctagcgc cggagagctt
     5101 caaaagggga acgaactcgc actaccgtct aaatacgtga atttcctgta tttagcgtcc
     5161 cattacgaga agttgaaagg ttcacctgaa gataacgaac agaagcaact ttttgttgag
     5221 cagcacaaac attatctcga cgaaatcata gagcaaattt cggaattcag taagagagtc
     5281 atcctagctg atgccaatct ggacaaagta ttaagcgcat acaacaagca cagggataaa
     5341 cccatacgtg agcaggcgga aaatattatc catttgttta ctcttaccaa cctcggcgct
     5401 ccagccgcat tcaagtattt tgacacaacg atagatcgca aacgatacac ttctaccaag
     5461 gaggtgctag acgcgacact gattcaccaa tccatcacgg gattatatga aactcggata
     5521 gatttgtcac agcttggggg tgactctggt ggttctacta atctgtcaga tattattgaa
     5581 aaggagaccg gtaagcaact ggttatccag gaatccatcc tcatgctccc agaggaggtg
     5641 gaagaagtca ttgggaacaa gccggaaagc gatatactcg tgcacaccgc ctacgacgag
     5701 agcaccgacg agaatgtcat gcttctgact agcgacgccc ctgaatacaa gccttgggct
     5761 ctggtcatac aggatagcaa cggtgagaac aagattaaga tgctctctgg tggttctccc
     5821 aagaagaaga ggaaagtcta accggtcatc atcaccatca ccattgagtt taaacccgct
     5881 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     5941 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     6001 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     6061 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     6121 ctgaggcgga aagaaccagc tggggctcga taccgtcgac ctctagctag agcttggcgt
     6181 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     6241 tacgagccgg aagcataaag tgtaaagcct agggtgccta atgagtgagc taactcacat
     6301 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     6361 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     6421 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     6481 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     6541 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6601 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6661 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6721 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6781 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6841 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6901 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6961 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     7021 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     7081 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     7141 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     7201 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     7261 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     7321 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     7381 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     7441 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     7501 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     7561 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     7621 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     7681 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     7741 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     7801 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     7861 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     7921 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     7981 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     8041 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     8101 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     8161 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     8221 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     8281 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     8341 acgtcgacgg atcgggagat cgatctcccg atcccctagg gtcgactctc agtacaatct
     8401 gctctgatgc cgcatagtta agccagtatc tgctccctgc tt
//
