LOCUS       sequence-271476-        8760 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses ABE8.20-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 Dec 15, 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: ABE8.20-d
FEATURES             Location/Qualifiers
     source          1..8760
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(133..152)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        324..703
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        704..907
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     858..878
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     949..968
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        949..967
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(981..1481,1482..1577,1578..2075,2076..2171)
                     /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
                     EREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLYDATLYST
                     FEPCVMCAGAMIHSRIGRVVFGVRNAKTGAAGSLMDVLHHPGMNHRVEITEGILADECA
                     ALLCRFFRMPRRVFNAQKKAQSSTDSGGSSGGSSGSETPGTSESATPESSGGSSGGS"
     CDS             join(2172..2195,2196..2198,2199..6272)
                     /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             2172..6272
                     /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             6318..6338
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6347..6364
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6387..6404)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6393..6617
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6688..6704)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6688..6704)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6701..6723)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6712..6728
                     /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(6736..6742,6743..6760,6761..6766))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6781..6802
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6919..6936)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7090..7678)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7170..7189)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7849..8640,8641..8709))
                     /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     8472..8491
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(8710..8760,1..54))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
       61 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat cgatctcccg atcccctagg
      121 gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
      181 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
      241 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
      301 tgtacgggcc agatatacgc gttgacattg attattgact agttattaat agtaatcaat
      361 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
      421 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
      481 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      541 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      601 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      661 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
      721 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      781 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      841 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      901 cagagctggt ttagtgaacc gtcagatccg ctagagatcc gcggccgcta atacgactca
      961 ctatagggag agccgccacc atgtccgaag tcgagttttc ccatgagtac tggatgagac
     1021 acgcattgac tctcgcaaag agggcttggg atgaacgcga ggtgcccgtg ggggcagtac
     1081 tcgtgcataa caatcgcgta atcggcgaag gttggaatag gccgatcgga cgccacgacc
     1141 ccactgcaca tgcggaaatc atggcccttc gacagggagg gcttgtgatg cagaattatc
     1201 gacttatcga tgcgacgctg tacgtcacgc ttgaaccttg cgtaatgtgc gcgggagcta
     1261 tgattcactc ccgcattgga cgagttgtat tcggtgcccg cgacgccaag acgggtgccg
     1321 caggttcact gatggacgtg ctgcatcacc caggcatgaa ccaccgggta gaaatcacag
     1381 aaggcatatt ggcggacgaa tgtgcggcgc tgttgtccga cttttttcgc atgcggaggc
     1441 aggagatcaa ggcccagaaa aaagcacaat cctctactga ctctggtggt tcttctggtg
     1501 gttctagcgg cagcgagact cccgggacct cagagtccgc cacacccgaa agttctggtg
     1561 gttcttctgg tggttcttcc gaagtcgagt tttcccatga gtactggatg agacacgcat
     1621 tgactctcgc aaagagggct cgagatgaac gcgaggtgcc cgtgggggca gtactcgtgc
     1681 tcaacaatcg cgtaatcggc gaaggttgga atagggcaat cggactccac gaccccactg
     1741 cacatgcgga aatcatggcc cttcgacagg gagggcttgt gatgcagaat tatcgacttt
     1801 atgatgcgac gctgtactcg acgtttgaac cttgcgtaat gtgcgcggga gctatgattc
     1861 actcccgcat tggacgagtt gtattcggtg ttcgcaacgc caagacgggt gccgcaggtt
     1921 cactgatgga cgtgctgcat catccaggca tgaaccaccg ggtagaaatc acagaaggca
     1981 tattggcgga cgaatgtgcg gcgctgttgt gtcgtttttt tcgcatgccc aggcgggtct
     2041 ttaacgccca gaaaaaagca caatcctcta ctgactctgg tggttcttct ggtggttcta
     2101 gcggcagcga gactcccggg acctcagagt ccgccacacc cgaaagttct ggtggttctt
     2161 ctggtggttc tgacaagaag tacagcatcg gcctggccat cggcaccaac tctgtgggct
     2221 gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca
     2281 ccgaccggca cagcatcaag aagaacctga tcggagccct gctgttcgac agcggcgaaa
     2341 cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga cggaagaacc
     2401 ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac gacagcttct
     2461 tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag cggcacccca
     2521 tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc atctaccacc
     2581 tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc tatctggccc
     2641 tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg aaccccgaca
     2701 acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag ctgttcgagg
     2761 aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc agactgagca
     2821 agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag aatggcctgt
     2881 tcggaaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc aacttcgacc
     2941 tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac ctggacaacc
     3001 tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag aacctgtccg
     3061 acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag gcccccctga
     3121 gcgcctctat gatcaagaga tacgacgagc accaccagga cctgaccctg ctgaaagctc
     3181 tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag agcaagaacg
     3241 gctacgccgg ctacattgac ggcggagcca gccaggaaga gttctacaag ttcatcaagc
     3301 ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac agagaggacc
     3361 tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc ccaccagatc cacctgggag
     3421 agctgcacgc cattctgcgg cggcaggaag atttttaccc attcctgaag gacaaccggg
     3481 aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct ctggccaggg
     3541 gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc ccctggaact
     3601 tcgaggaagt ggtggacaag ggcgcttccg cccagagctt catcgagcgg atgaccaact
     3661 tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg tacgagtact
     3721 tcaccgtgta taacgagctg accaaagtga aatacgtgac cgagggaatg agaaagcccg
     3781 ccttcctgag cggcgagcag aaaaaggcca tcgtggacct gctgttcaag accaaccgga
     3841 aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc ttcgactccg
     3901 tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac cacgatctgc
     3961 tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac attctggaag
     4021 atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa cggctgaaaa
     4081 cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggcgg agatacaccg
     4141 gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag tccggcaaga
     4201 caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg cagctgatcc
     4261 acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg
     4321 atagcctgca cgagcacatt gccaatctgg ccggcagccc cgccattaag aagggcatcc
     4381 tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat gggccggcac aagcccgaga
     4441 acatcgtgat cgaaatggcc agagagaacc agaccaccca gaagggacag aagaacagcc
     4501 gcgagagaat gaagcggatc gaagagggca tcaaagagct gggcagccag atcctgaaag
     4561 aacaccccgt ggaaaacacc cagctgcaga acgagaagct gtacctgtac tacctgcaga
     4621 atgggcggga tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc gactacgatg
     4681 tggaccatat cgtgcctcag agctttctga aggacgactc catcgacaac aaggtgctga
     4741 ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga
     4801 agatgaagaa ctactggcgg cagctgctga acgccaagct gattacccag agaaagttcg
     4861 acaatctgac caaggccgag agaggcggcc tgagcgaact ggataaggcc ggcttcatca
     4921 agagacagct ggtggaaacc cggcagatca caaagcacgt ggcacagatc ctggactccc
     4981 ggatgaacac taagtacgac gagaatgaca agctgatccg ggaagtgaaa gtgatcaccc
     5041 tgaagtccaa gctggtgtcc gatttccgga aggatttcca gttttacaaa gtgcgcgaga
     5101 tcaacaacta ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga accgccctga
     5161 tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg
     5221 tgcggaagat gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc aagtacttct
     5281 tctacagcaa catcatgaac tttttcaaga ccgagattac cctggccaac ggcgagatcc
     5341 ggaagcggcc tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg gataagggcc
     5401 gggattttgc caccgtgcgg aaagtgctga gcatgcccca agtgaatatc gtgaaaaaga
     5461 ccgaggtgca gacaggcggc ttcagcaaag agtctatcct gcccaagagg aacagcgata
     5521 agctgatcgc cagaaagaag gactgggacc ctaagaagta cggcggcttc gacagcccca
     5581 ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc aagaaactga
     5641 agagtgtgaa agagctgctg gggatcacca tcatggaaag aagcagcttc gagaagaatc
     5701 ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg atcatcaagc
     5761 tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg gcctctgccg
     5821 gcgaactgca gaagggaaac gaactggccc tgccctccaa atatgtgaac ttcctgtacc
     5881 tggccagcca ctatgagaag ctgaagggct cccccgagga taatgagcag aaacagctgt
     5941 ttgtggaaca gcacaagcac tacctggacg agatcatcga gcagatcagc gagttctcca
     6001 agagagtgat cctggccgac gctaatctgg acaaagtgct gtccgcctac aacaagcacc
     6061 gggataagcc catcagagag caggccgaga atatcatcca cctgtttacc ctgaccaatc
     6121 tgggagcccc tgccgccttc aagtactttg acaccaccat cgaccggaag aggtacacca
     6181 gcaccaaaga ggtgctggac gccaccctga tccaccagag catcaccggc ctgtacgaga
     6241 cacggatcga cctgtctcag ctgggaggtg acgagggagc tgataagcgc accgccgatg
     6301 gttccgagtt cgaaagcccc aagaagaaga ggaaagtcta accggtcatc atcaccatca
     6361 ccattgagtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     6421 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     6481 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     6541 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     6601 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcga taccgtcgac
     6661 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     6721 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct agggtgccta
     6781 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     6841 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6901 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6961 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     7021 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     7081 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     7141 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     7201 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     7261 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     7321 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     7381 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     7441 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     7501 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     7561 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     7621 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     7681 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     7741 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     7801 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     7861 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     7921 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     7981 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     8041 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     8101 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     8161 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     8221 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     8281 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     8341 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     8401 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     8461 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     8521 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     8581 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     8641 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     8701 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
//
