LOCUS       sequence-224189-        8442 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses hA3A-BE3 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 Apr 14, 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
FEATURES             Location/Qualifiers
     source          1..8442
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        124..503
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        504..707
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     658..678
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     749..768
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        749..767
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      775..784
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             781..1377
                     /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
                     GCAGEVRAFLQENTHVRLRIFAARIYDYDPLYKEALQMLRDAGAQVSIMTYDEFKHCWD
                     TFVDHQGCPFQPWDGLDEHSQALSGRLRAILQNQGN"
     CDS             join(1426..4824,4825..4827,4828..5424,5425,5426..5428,
                     5429..5430,5431..5433,5434..5526)
                     /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(1426..1449,1450..1452,1453..5526)
                     /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             5539..5787
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5800..5820
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5829..5846
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5869..5886)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5875..6099
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6170..6186)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6170..6186)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6183..6205)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6194..6210
                     /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(6218..6224,6225..6242,6243..6248))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6263..6284
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6401..6418)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6572..7160)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6652..6671)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7331..8122,8123..8191))
                     /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     7954..7973
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8192..8296)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8375..8394)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
       61 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
      121 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
      181 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      241 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      301 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      361 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      421 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      481 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      541 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      601 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      661 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc
      721 gtcagatccg ctagagatcc gcggccgcta atacgactca ctatagggag agccgccacc
      781 atggaagcca gcccagcatc cgggcccaga cacttgatgg atccacacat attcacttcc
      841 aactttaaca atggcattgg aaggcataag acctacctgt gctacgaagt ggagcgcctg
      901 gacaatggca cctcggtcaa gatggaccag cacaggggct ttctacacaa ccaggctaag
      961 aatcttctct gtggctttta cggccgccat gcggagctgc gcttcttgga cctggttcct
     1021 tctttgcagt tggacccggc ccagatctac agggtcactt ggttcatctc ctggagcccc
     1081 tgcttctcct ggggctgtgc cggggaagtg cgtgcgttcc ttcaggagaa cacacacgtg
     1141 agactgcgta tcttcgctgc ccgcatctat gattacgacc ccctatataa ggaggcactg
     1201 caaatgctgc gggatgctgg ggcccaagtc tccatcatga cctacgatga atttaagcac
     1261 tgctgggaca cctttgtgga ccaccaggga tgtcccttcc agccctggga tggactagat
     1321 gagcacagcc aagccctgag tgggaggctg cgggccattc tccagaatca gggaaacagc
     1381 ggcagcgaga ctcccgggac ctcagagtcc gccacacccg aaagtgataa aaagtattct
     1441 attggtttag ccatcggcac taattccgtt ggatgggctg tcataaccga tgaatacaaa
     1501 gtaccttcaa agaaatttaa ggtgttgggg aacacagacc gtcattcgat taaaaagaat
     1561 cttatcggtg ccctcctatt cgatagtggc gaaacggcag aggcgactcg cctgaaacga
     1621 accgctcgga gaaggtatac acgtcgcaag aaccgaatat gttacttaca agaaattttt
     1681 agcaatgaga tggccaaagt tgacgattct ttctttcacc gtttggaaga gtccttcctt
     1741 gtcgaagagg acaagaaaca tgaacggcac cccatctttg gaaacatagt agatgaggtg
     1801 gcatatcatg aaaagtaccc aacgatttat cacctcagaa aaaagctagt tgactcaact
     1861 gataaagcgg acctgaggtt aatctacttg gctcttgccc atatgataaa gttccgtggg
     1921 cactttctca ttgagggtga tctaaatccg gacaactcgg atgtcgacaa actgttcatc
     1981 cagttagtac aaacctataa tcagttgttt gaagagaacc ctataaatgc aagtggcgtg
     2041 gatgcgaagg ctattcttag cgcccgcctc tctaaatccc gacggctaga aaacctgatc
     2101 gcacaattac ccggagagaa gaaaaatggg ttgttcggta accttatagc gctctcacta
     2161 ggcctgacac caaattttaa gtcgaacttc gacttagctg aagatgccaa attgcagctt
     2221 agtaaggaca cgtacgatga cgatctcgac aatctactgg cacaaattgg agatcagtat
     2281 gcggacttat ttttggctgc caaaaacctt agcgatgcaa tcctcctatc tgacatactg
     2341 agagttaata ctgagattac caaggcgccg ttatccgctt caatgatcaa aaggtacgat
     2401 gaacatcacc aagacttgac acttctcaag gccctagtcc gtcagcaact gcctgagaaa
     2461 tataaggaaa tattctttga tcagtcgaaa aacgggtacg caggttatat tgacggcgga
     2521 gcgagtcaag aggaattcta caagtttatc aaacccatat tagagaagat ggatgggacg
     2581 gaagagttgc ttgtaaaact caatcgcgaa gatctactgc gaaagcagcg gactttcgac
     2641 aacggtagca ttccacatca aatccactta ggcgaattgc atgctatact tagaaggcag
     2701 gaggattttt atccgttcct caaagacaat cgtgaaaaga ttgagaaaat cctaaccttt
     2761 cgcatacctt actatgtggg acccctggcc cgagggaact ctcggttcgc atggatgaca
     2821 agaaagtccg aagaaacgat tactccatgg aattttgagg aagttgtcga taaaggtgcg
     2881 tcagctcaat cgttcatcga gaggatgacc aactttgaca agaatttacc gaacgaaaaa
     2941 gtattgccta agcacagttt actttacgag tatttcacag tgtacaatga actcacgaaa
     3001 gttaagtatg tcactgaggg catgcgtaaa cccgcctttc taagcggaga acagaagaaa
     3061 gcaatagtag atctgttatt caagaccaac cgcaaagtga cagttaagca attgaaagag
     3121 gactacttta agaaaattga atgcttcgat tctgtcgaga tctccggggt agaagatcga
     3181 tttaatgcgt cacttggtac gtatcatgac ctcctaaaga taattaaaga taaggacttc
     3241 ctggataacg aagagaatga agatatctta gaagatatag tgttgactct taccctcttt
     3301 gaagatcggg aaatgattga ggaaagacta aaaacatacg ctcacctgtt cgacgataag
     3361 gttatgaaac agttaaagag gcgtcgctat acgggctggg gacgattgtc gcggaaactt
     3421 atcaacggga taagagacaa gcaaagtggt aaaactattc tcgattttct aaagagcgac
     3481 ggcttcgcca ataggaactt tatgcagctg atccatgatg actctttaac cttcaaagag
     3541 gatatacaaa aggcacaggt ttccggacaa ggggactcat tgcacgaaca tattgcgaat
     3601 cttgctggtt cgccagccat caaaaagggc atactccaga cagtcaaagt agtggatgag
     3661 ctagttaagg tcatgggacg tcacaaaccg gaaaacattg taatcgagat ggcacgcgaa
     3721 aatcaaacga ctcagaaggg gcaaaaaaac agtcgagagc ggatgaagag aatagaagag
     3781 ggtattaaag aactgggcag ccagatctta aaggagcatc ctgtggaaaa tacccaattg
     3841 cagaacgaga aactttacct ctattaccta caaaatggaa gggacatgta tgttgatcag
     3901 gaactggaca taaaccgttt atctgattac gacgtcgatc acattgtacc ccaatccttt
     3961 ttgaaggacg attcaatcga caataaagtg cttacacgct cggataagaa ccgagggaaa
     4021 agtgacaatg ttccaagcga ggaagtcgta aagaaaatga agaactattg gcggcagctc
     4081 ctaaatgcga aactgataac gcaaagaaag ttcgataact taactaaagc tgagaggggt
     4141 ggcttgtctg aacttgacaa ggccggattt attaaacgtc agctcgtgga aacccgccaa
     4201 atcacaaagc atgttgcaca gatactagat tcccgaatga atacgaaata cgacgagaac
     4261 gataagctga ttcgggaagt caaagtaatc actttaaagt caaaattggt gtcggacttc
     4321 agaaaggatt ttcaattcta taaagttagg gagataaata actaccacca tgcgcacgac
     4381 gcttatctta atgccgtcgt agggaccgca ctcattaaga aatacccgaa gctagaaagt
     4441 gagtttgtgt atggtgatta caaagtttat gacgtccgta agatgatcgc gaaaagcgaa
     4501 caggagatag gcaaggctac agccaaatac ttcttttatt ctaacattat gaatttcttt
     4561 aagacggaaa tcactctggc aaacggagag atacgcaaac gacctttaat tgaaaccaat
     4621 ggggagacag gtgaaatcgt atgggataag ggccgggact tcgcgacggt gagaaaagtt
     4681 ttgtccatgc cccaagtcaa catagtaaag aaaactgagg tgcagaccgg agggttttca
     4741 aaggaatcga ttcttccaaa aaggaatagt gataagctca tcgctcgtaa aaaggactgg
     4801 gacccgaaaa agtacggtgg cttcgatagc cctacagttg cctattctgt cctagtagtg
     4861 gcaaaagttg agaagggaaa atccaagaaa ctgaagtcag tcaaagaatt attggggata
     4921 acgattatgg agcgctcgtc ttttgaaaag aaccccatcg acttccttga ggcgaaaggt
     4981 tacaaggaag taaaaaagga tctcataatt aaactaccaa agtatagtct gtttgagtta
     5041 gaaaatggcc gaaaacggat gttggctagc gccggagagc ttcaaaaggg gaacgaactc
     5101 gcactaccgt ctaaatacgt gaatttcctg tatttagcgt cccattacga gaagttgaaa
     5161 ggttcacctg aagataacga acagaagcaa ctttttgttg agcagcacaa acattatctc
     5221 gacgaaatca tagagcaaat ttcggaattc agtaagagag tcatcctagc tgatgccaat
     5281 ctggacaaag tattaagcgc atacaacaag cacagggata aacccatacg tgagcaggcg
     5341 gaaaatatta tccatttgtt tactcttacc aacctcggcg ctccagccgc attcaagtat
     5401 tttgacacaa cgatagatcg caaacgatac acttctacca aggaggtgct agacgcgaca
     5461 ctgattcacc aatccatcac gggattatat gaaactcgga tagatttgtc acagcttggg
     5521 ggtgactctg gtggttctac taatctgtca gatattattg aaaaggagac cggtaagcaa
     5581 ctggttatcc aggaatccat cctcatgctc ccagaggagg tggaagaagt cattgggaac
     5641 aagccggaaa gcgatatact cgtgcacacc gcctacgacg agagcaccga cgagaatgtc
     5701 atgcttctga ctagcgacgc ccctgaatac aagccttggg ctctggtcat acaggatagc
     5761 aacggtgaga acaagattaa gatgctctct ggtggttctc ccaagaagaa gaggaaagtc
     5821 taaccggtca tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc
     5881 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     5941 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     6001 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     6061 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     6121 gctggggctc gataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     6181 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     6241 agtgtaaagc ctagggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     6301 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     6361 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     6421 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     6481 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     6541 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6601 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6661 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6721 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6781 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6841 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6901 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6961 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     7021 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     7081 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     7141 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     7201 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     7261 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     7321 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     7381 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     7441 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     7501 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     7561 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     7621 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     7681 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     7741 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7801 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7861 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7921 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7981 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     8041 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     8101 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     8161 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     8221 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     8281 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     8341 atcgatctcc cgatccccta gggtcgactc tcagtacaat ctgctctgat gccgcatagt
     8401 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tg
//
