LOCUS       sequence-269979-        8760 bp    DNA     circular SYN 14-APR-2025
DEFINITION  expresses ABE8.17-d in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8760)
  AUTHORS   Gaudelli NM, Lam DK, Rees HA, Sola-Esteves NM, Barrera LA, Born DA, 
            Edwards A, Gehrke JM, Lee SJ, Liquori AJ, Murray R, Packer MS, 
            Rinaldi C, Slaymaker IM, Yen J, Young LE, Ciaramella G
  TITLE     Directed evolution of adenine base editors with increased activity 
            and therapeutic application.
  JOURNAL   Nat Biotechnol. 2020 Apr 13. pii: 10.1038/s41587-020-0491-6. doi: 
            10.1038/s41587-020-0491-6.
  PUBMED    32284586
REFERENCE   2  (bases 1 to 8760)
  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. 2020 Apr 13. pii: 10.1038/s41587-020-0491-6. doi: 
            10.1038/s41587-020-0491-6."
COMMENT     Sequence Label: ABE8.17-d
FEATURES             Location/Qualifiers
     source          1..8760
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     16..35
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        16..34
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(48..548,549..644,645..1142,1143..1238)
                     /codon_start=1
                     /product="adenine base editor comprising wild-type E. coli 
                     TadA fused to the evolved TadA*(7.10) mutant (Gaudelli et 
                     al., 2017)"
                     /label=ABE(7.10)
                     /translation="MSEVEFSHEYWMRHALTLAKRAWDEREVPVGAVLVHNNRVIGEGW
                     NRPIGRHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTLEPCVMCAGAMIHSRIGRVVF
                     GARDAKTGAAGSLMDVLHHPGMNHRVEITEGILADECAALLSDFFRMRRQEIKAQKKAQ
                     SSTDSGGSSGGSSGSETPGTSESATPESSGGSSGGSSEVEFSHEYWMRHALTLAKRARD
                     EREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYST
                     FEPCVMCAGAMIHSRIGRVVFGVRNAKTGAAGSLMDVLHYPGMNHRVEITEGILADECA
                     ALLCYFFRMPRRVFNAQKKAQSSTDSGGSSGGSSGSETPGTSESATPESSGGSSGGS"
     CDS             join(1239..1262,1263..1265,1266..5339)
                     /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             1239..5339
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5385..5405
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5414..5431
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5454..5471)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5460..5684
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5755..5771)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5755..5771)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5768..5790)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5779..5795
                     /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(5803..5809,5810..5827,5828..5833))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5848..5869
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5986..6003)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6157..6745)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6237..6256)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6916..7707,7708..7776))
                     /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     7539..7558
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7777..7881)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7960..7979)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        8151..8530
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8531..8734
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8685..8705
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 gagatccgcg gccgctaata cgactcacta tagggagagc cgccaccatg tccgaagtcg
       61 agttttccca tgagtactgg atgagacacg cattgactct cgcaaagagg gcttgggatg
      121 aacgcgaggt gcccgtgggg gcagtactcg tgcataacaa tcgcgtaatc ggcgaaggtt
      181 ggaataggcc gatcggacgc cacgacccca ctgcacatgc ggaaatcatg gcccttcgac
      241 agggagggct tgtgatgcag aattatcgac ttatcgatgc gacgctgtac gtcacgcttg
      301 aaccttgcgt aatgtgcgcg ggagctatga ttcactcccg cattggacga gttgtattcg
      361 gtgcccgcga cgccaagacg ggtgccgcag gttcactgat ggacgtgctg catcacccag
      421 gcatgaacca ccgggtagaa atcacagaag gcatattggc ggacgaatgt gcggcgctgt
      481 tgtccgactt ttttcgcatg cggaggcagg agatcaaggc ccagaaaaaa gcacaatcct
      541 ctactgactc tggtggttct tctggtggtt ctagcggcag cgagactccc gggacctcag
      601 agtccgccac acccgaaagt tctggtggtt cttctggtgg ttcttccgaa gtcgagtttt
      661 cccatgagta ctggatgaga cacgcattga ctctcgcaaa gagggctcga gatgaacgcg
      721 aggtgcccgt gggggcagta ctcgtgctca acaatcgcgt aatcggcgaa ggttggaata
      781 gggcaatcgg actccacgac cccactgcac atgcggaaat catggccctt cgacagggag
      841 ggcttgtgat gcagaattat cgacttatcg atgcgacgct gtactcgacg tttgaacctt
      901 gcgtaatgtg cgcgggagct atgattcact cccgcattgg acgagttgta ttcggtgttc
      961 gcaacgccaa gacgggtgcc gcaggttcac tgatggacgt gctgcattac ccaggcatga
     1021 accaccgggt agaaatcaca gaaggcatat tggcggacga atgtgcggcg ctgttgtgtt
     1081 acttttttcg catgcccagg cgtgtcttta acgcccagaa aaaagcacaa tcctctactg
     1141 actctggtgg ttcttctggt ggttctagcg gcagcgagac tcccgggacc tcagagtccg
     1201 ccacacccga aagttctggt ggttcttctg gtggttctga caagaagtac agcatcggcc
     1261 tggccatcgg caccaactct gtgggctggg ccgtgatcac cgacgagtac aaggtgccca
     1321 gcaagaaatt caaggtgctg ggcaacaccg accggcacag catcaagaag aacctgatcg
     1381 gagccctgct gttcgacagc ggcgaaacag ccgaggccac ccggctgaag agaaccgcca
     1441 gaagaagata caccagacgg aagaaccgga tctgctatct gcaagagatc ttcagcaacg
     1501 agatggccaa ggtggacgac agcttcttcc acagactgga agagtccttc ctggtggaag
     1561 aggataagaa gcacgagcgg caccccatct tcggcaacat cgtggacgag gtggcctacc
     1621 acgagaagta ccccaccatc taccacctga gaaagaaact ggtggacagc accgacaagg
     1681 ccgacctgcg gctgatctat ctggccctgg cccacatgat caagttccgg ggccacttcc
     1741 tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc atccagctgg
     1801 tgcagaccta caaccagctg ttcgaggaaa accccatcaa cgccagcggc gtggacgcca
     1861 aggccatcct gtctgccaga ctgagcaaga gcagacggct ggaaaatctg atcgcccagc
     1921 tgcccggcga gaagaagaat ggcctgttcg gaaacctgat tgccctgagc ctgggcctga
     1981 cccccaactt caagagcaac ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg
     2041 acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag tacgccgacc
     2101 tgtttctggc cgccaagaac ctgtccgacg ccatcctgct gagcgacatc ctgagagtga
     2161 acaccgagat caccaaggcc cccctgagcg cctctatgat caagagatac gacgagcacc
     2221 accaggacct gaccctgctg aaagctctcg tgcggcagca gctgcctgag aagtacaaag
     2281 agattttctt cgaccagagc aagaacggct acgccggcta cattgacggc ggagccagcc
     2341 aggaagagtt ctacaagttc atcaagccca tcctggaaaa gatggacggc accgaggaac
     2401 tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca gcggaccttc gacaacggca
     2461 gcatccccca ccagatccac ctgggagagc tgcacgccat tctgcggcgg caggaagatt
     2521 tttacccatt cctgaaggac aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc
     2581 cctactacgt gggccctctg gccaggggaa acagcagatt cgcctggatg accagaaaga
     2641 gcgaggaaac catcaccccc tggaacttcg aggaagtggt ggacaagggc gcttccgccc
     2701 agagcttcat cgagcggatg accaacttcg ataagaacct gcccaacgag aaggtgctgc
     2761 ccaagcacag cctgctgtac gagtacttca ccgtgtataa cgagctgacc aaagtgaaat
     2821 acgtgaccga gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg
     2881 tggacctgct gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa gaggactact
     2941 tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg cgtggaagat cggttcaacg
     3001 cctccctggg cacataccac gatctgctga aaattatcaa ggacaaggac ttcctggaca
     3061 atgaggaaaa cgaggacatt ctggaagata tcgtgctgac cctgacactg tttgaggaca
     3121 gagagatgat cgaggaacgg ctgaaaacct atgcccacct gttcgacgac aaagtgatga
     3181 agcagctgaa gcggcggaga tacaccggct ggggcaggct gagccggaag ctgatcaacg
     3241 gcatccggga caagcagtcc ggcaagacaa tcctggattt cctgaagtcc gacggcttcg
     3301 ccaacagaaa cttcatgcag ctgatccacg acgacagcct gacctttaaa gaggacatcc
     3361 agaaagccca ggtgtccggc cagggcgata gcctgcacga gcacattgcc aatctggccg
     3421 gcagccccgc cattaagaag ggcatcctgc agacagtgaa ggtggtggac gagctcgtga
     3481 aagtgatggg ccggcacaag cccgagaaca tcgtgatcga aatggccaga gagaaccaga
     3541 ccacccagaa gggacagaag aacagccgcg agagaatgaa gcggatcgaa gagggcatca
     3601 aagagctggg cagccagatc ctgaaagaac accccgtgga aaacacccag ctgcagaacg
     3661 agaagctgta cctgtactac ctgcagaatg ggcgggatat gtacgtggac caggaactgg
     3721 acatcaaccg gctgtccgac tacgatgtgg accatatcgt gcctcagagc tttctgaagg
     3781 acgactccat cgacaacaag gtgctgacca gaagcgacaa gaaccggggc aagagcgaca
     3841 acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg
     3901 ccaagctgat tacccagaga aagttcgaca atctgaccaa ggccgagaga ggcggcctga
     3961 gcgaactgga taaggccggc ttcatcaaga gacagctggt ggaaacccgg cagatcacaa
     4021 agcacgtggc acagatcctg gactcccgga tgaacactaa gtacgacgag aatgacaagc
     4081 tgatccggga agtgaaagtg atcaccctga agtccaagct ggtgtccgat ttccggaagg
     4141 atttccagtt ttacaaagtg cgcgagatca acaactacca ccacgcccac gacgcctacc
     4201 tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc taagctggaa agcgagttcg
     4261 tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa
     4321 tcggcaaggc taccgccaag tacttcttct acagcaacat catgaacttt ttcaagaccg
     4381 agattaccct ggccaacggc gagatccgga agcggcctct gatcgagaca aacggcgaaa
     4441 ccggggagat cgtgtgggat aagggccggg attttgccac cgtgcggaaa gtgctgagca
     4501 tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc agcaaagagt
     4561 ctatcctgcc caagaggaac agcgataagc tgatcgccag aaagaaggac tgggacccta
     4621 agaagtacgg cggcttcgac agccccaccg tggcctattc tgtgctggtg gtggccaaag
     4681 tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga gctgctgggg atcaccatca
     4741 tggaaagaag cagcttcgag aagaatccca tcgactttct ggaagccaag ggctacaaag
     4801 aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc cctgttcgag ctggaaaacg
     4861 gccggaagag aatgctggcc tctgccggcg aactgcagaa gggaaacgaa ctggccctgc
     4921 cctccaaata tgtgaacttc ctgtacctgg ccagccacta tgagaagctg aagggctccc
     4981 ccgaggataa tgagcagaaa cagctgtttg tggaacagca caagcactac ctggacgaga
     5041 tcatcgagca gatcagcgag ttctccaaga gagtgatcct ggccgacgct aatctggaca
     5101 aagtgctgtc cgcctacaac aagcaccggg ataagcccat cagagagcag gccgagaata
     5161 tcatccacct gtttaccctg accaatctgg gagcccctgc cgccttcaag tactttgaca
     5221 ccaccatcga ccggaagagg tacaccagca ccaaagaggt gctggacgcc accctgatcc
     5281 accagagcat caccggcctg tacgagacac ggatcgacct gtctcagctg ggaggtgacg
     5341 agggagctga taagcgcacc gccgatggtt ccgagttcga aagccccaag aagaagagga
     5401 aagtctaacc ggtcatcatc accatcacca ttgagtttaa acccgctgat cagcctcgac
     5461 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     5521 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     5581 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     5641 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     5701 aaccagctgg ggctcgatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     5761 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     5821 cataaagtgt aaagcctagg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     5881 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     5941 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     6001 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     6061 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6121 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6181 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6241 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     6301 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     6361 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     6421 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     6481 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6541 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     6601 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6661 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6721 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6781 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6841 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6901 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6961 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     7021 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     7081 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7141 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7201 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     7261 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     7321 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     7381 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7441 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7501 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     7561 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7621 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7681 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7741 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7801 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7861 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     7921 gggagatcga tctcccgatc ccctagggtc gactctcagt acaatctgct ctgatgccgc
     7981 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag
     8041 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag
     8101 ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt
     8161 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     8221 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     8281 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     8341 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     8401 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     8461 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     8521 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     8581 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     8641 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     8701 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
//
