LOCUS       sequence-379979-        8778 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express TadCBEb (with SpCas9) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8778)
  AUTHORS   Neugebauer ME, Hsu A, Arbab M, Krasnow NA, McElroy AN, Pandey S, 
            Doman JL, Huang TP, Raguram A, Banskota S, Newby GA, Tolar J, Osborn
            MJ, Liu DR
  TITLE     Evolution of an adenine base editor into a small, efficient cytosine
            base editor with low off-target activity.
  JOURNAL   Nat Biotechnol. 2022 Nov 10. pii: 10.1038/s41587-022-01533-6. doi: 
            10.1038/s41587-022-01533-6.
  PUBMED    36357719
REFERENCE   2  (bases 1 to 8778)
  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. 2022 Nov 10. pii: 10.1038/s41587-022-01533-6. doi: 
            10.1038/s41587-022-01533-6."
COMMENT     Sequence Label: SpCas9 TadCBEb
FEATURES             Location/Qualifiers
     source          1..8778
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        8..387
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        388..591
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     542..562
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     633..652
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        633..651
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             701..721
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1319..1342,1343..1345,1346..5419)
                     /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             1319..5419
                     /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             5450..5698
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5729..5977
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6020..6040
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6049..6066
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6089..6106)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6095..6319
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6390..6406)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6390..6406)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6403..6425)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6414..6430
                     /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(6438..6444,6445..6462,6463..6468))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6483..6504
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6621..6638)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6792..7380)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6872..6891)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7551..8342,8343..8411))
                     /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     8174..8193
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8412..8516)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8595..8614)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
       61 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      121 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      181 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      241 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      301 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      361 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
      421 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
      481 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
      541 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg
      601 aaccgtcaga tccgctagag atccgcggcc gctaatacga ctcactatag ggagagccgc
      661 caccatgaaa cggacagccg acggaagcga gttcgagtca ccaaagaaga agcggaaagt
      721 cagttctgag gtggagtttt cccacgagta ctggatgaga catgccctga ccctggccaa
      781 gagggcacgg gatgaggggg cggggcctgt gggagccgtg ctggtgctga acaatagagt
      841 gatcggcgag ggctggaaca gagccatcgg cctgcacgac ccaacagccc atgccgaaat
      901 tatggccctg agacagggcg gcctggtcat gcagaactac agactgtttg acgccaccct
      961 gtacgtgaca ttcgagcctt gcgtgatgtg cgccggcgcc atgatcaact ctaggatcgg
     1021 ccgcgtggtg tttggcgtga ggaactcaaa aagaggcgcc gcaggctccc tgatgaacgt
     1081 gctgaactac cccggcatga atcaccgcgt cgaaattacc gagggaatcc tggcagatga
     1141 atgtgccgcc ctgctgtgcg atttctatcg gatgcctaga cgggtgttca attctcagaa
     1201 gaaggcccag agctccatca actctggcgg atctagcgga ggatcctctg gcagcgagac
     1261 accaggaaca agcgagtcag caacaccaga gagcagtggc ggcagcagcg gcggcagcga
     1321 caagaagtac agcatcggcc tggccatcgg caccaactct gtgggctggg ccgtgatcac
     1381 cgacgagtac aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg accggcacag
     1441 catcaagaag aacctgatcg gagccctgct gttcgacagc ggcgaaacag ccgaggccac
     1501 ccggctgaag agaaccgcca gaagaagata caccagacgg aagaaccgga tctgctatct
     1561 gcaagagatc ttcagcaacg agatggccaa ggtggacgac agcttcttcc acagactgga
     1621 agagtccttc ctggtggaag aggataagaa gcacgagcgg caccccatct tcggcaacat
     1681 cgtggacgag gtggcctacc acgagaagta ccccaccatc taccacctga gaaagaaact
     1741 ggtggacagc accgacaagg ccgacctgcg gctgatctat ctggccctgg cccacatgat
     1801 caagttccgg ggccacttcc tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga
     1861 caagctgttc atccagctgg tgcagaccta caaccagctg ttcgaggaaa accccatcaa
     1921 cgccagcggc gtggacgcca aggccatcct gtctgccaga ctgagcaaga gcagacggct
     1981 ggaaaatctg atcgcccagc tgcccggcga gaagaagaat ggcctgttcg gaaacctgat
     2041 tgccctgagc ctgggcctga cccccaactt caagagcaac ttcgacctgg ccgaggatgc
     2101 caaactgcag ctgagcaagg acacctacga cgacgacctg gacaacctgc tggcccagat
     2161 cggcgaccag tacgccgacc tgtttctggc cgccaagaac ctgtccgacg ccatcctgct
     2221 gagcgacatc ctgagagtga acaccgagat caccaaggcc cccctgagcg cctctatgat
     2281 caagagatac gacgagcacc accaggacct gaccctgctg aaagctctcg tgcggcagca
     2341 gctgcctgag aagtacaaag agattttctt cgaccagagc aagaacggct acgccggcta
     2401 cattgacggc ggagccagcc aggaagagtt ctacaagttc atcaagccca tcctggaaaa
     2461 gatggacggc accgaggaac tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca
     2521 gcggaccttc gacaacggca gcatccccca ccagatccac ctgggagagc tgcacgccat
     2581 tctgcggcgg caggaagatt tttacccatt cctgaaggac aaccgggaaa agatcgagaa
     2641 gatcctgacc ttccgcatcc cctactacgt gggccctctg gccaggggaa acagcagatt
     2701 cgcctggatg accagaaaga gcgaggaaac catcaccccc tggaacttcg aggaagtggt
     2761 ggacaagggc gcttccgccc agagcttcat cgagcggatg accaacttcg ataagaacct
     2821 gcccaacgag aaggtgctgc ccaagcacag cctgctgtac gagtacttca ccgtgtataa
     2881 cgagctgacc aaagtgaaat acgtgaccga gggaatgaga aagcccgcct tcctgagcgg
     2941 cgagcagaaa aaggccatcg tggacctgct gttcaagacc aaccggaaag tgaccgtgaa
     3001 gcagctgaaa gaggactact tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg
     3061 cgtggaagat cggttcaacg cctccctggg cacataccac gatctgctga aaattatcaa
     3121 ggacaaggac ttcctggaca atgaggaaaa cgaggacatt ctggaagata tcgtgctgac
     3181 cctgacactg tttgaggaca gagagatgat cgaggaacgg ctgaaaacct atgcccacct
     3241 gttcgacgac aaagtgatga agcagctgaa gcggcggaga tacaccggct ggggcaggct
     3301 gagccggaag ctgatcaacg gcatccggga caagcagtcc ggcaagacaa tcctggattt
     3361 cctgaagtcc gacggcttcg ccaacagaaa cttcatgcag ctgatccacg acgacagcct
     3421 gacctttaaa gaggacatcc agaaagccca ggtgtccggc cagggcgata gcctgcacga
     3481 gcacattgcc aatctggccg gcagccccgc cattaagaag ggcatcctgc agacagtgaa
     3541 ggtggtggac gagctcgtga aagtgatggg ccggcacaag cccgagaaca tcgtgatcga
     3601 aatggccaga gagaaccaga ccacccagaa gggacagaag aacagccgcg agagaatgaa
     3661 gcggatcgaa gagggcatca aagagctggg cagccagatc ctgaaagaac accccgtgga
     3721 aaacacccag ctgcagaacg agaagctgta cctgtactac ctgcagaatg ggcgggatat
     3781 gtacgtggac caggaactgg acatcaaccg gctgtccgac tacgatgtgg accatatcgt
     3841 gcctcagagc tttctgaagg acgactccat cgacaacaag gtgctgacca gaagcgacaa
     3901 gaaccggggc aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta
     3961 ctggcggcag ctgctgaacg ccaagctgat tacccagaga aagttcgaca atctgaccaa
     4021 ggccgagaga ggcggcctga gcgaactgga taaggccggc ttcatcaaga gacagctggt
     4081 ggaaacccgg cagatcacaa agcacgtggc acagatcctg gactcccgga tgaacactaa
     4141 gtacgacgag aatgacaagc tgatccggga agtgaaagtg atcaccctga agtccaagct
     4201 ggtgtccgat ttccggaagg atttccagtt ttacaaagtg cgcgagatca acaactacca
     4261 ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc
     4321 taagctggaa agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat
     4381 cgccaagagc gagcaggaaa tcggcaaggc taccgccaag tacttcttct acagcaacat
     4441 catgaacttt ttcaagaccg agattaccct ggccaacggc gagatccgga agcggcctct
     4501 gatcgagaca aacggcgaaa ccggggagat cgtgtgggat aagggccggg attttgccac
     4561 cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac
     4621 aggcggcttc agcaaagagt ctatcctgcc caagaggaac agcgataagc tgatcgccag
     4681 aaagaaggac tgggacccta agaagtacgg cggcttcgac agccccaccg tggcctattc
     4741 tgtgctggtg gtggccaaag tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga
     4801 gctgctgggg atcaccatca tggaaagaag cagcttcgag aagaatccca tcgactttct
     4861 ggaagccaag ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc
     4921 cctgttcgag ctggaaaacg gccggaagag aatgctggcc tctgccggcg aactgcagaa
     4981 gggaaacgaa ctggccctgc cctccaaata tgtgaacttc ctgtacctgg ccagccacta
     5041 tgagaagctg aagggctccc ccgaggataa tgagcagaaa cagctgtttg tggaacagca
     5101 caagcactac ctggacgaga tcatcgagca gatcagcgag ttctccaaga gagtgatcct
     5161 ggccgacgct aatctggaca aagtgctgtc cgcctacaac aagcaccggg ataagcccat
     5221 cagagagcag gccgagaata tcatccacct gtttaccctg accaatctgg gagcccctgc
     5281 cgccttcaag tactttgaca ccaccatcga ccggaagagg tacaccagca ccaaagaggt
     5341 gctggacgcc accctgatcc accagagcat caccggcctg tacgagacac ggatcgacct
     5401 gtctcagctg ggaggtgaca gcggcgggag cggcgggagc ggggggagca ctaatctgag
     5461 cgacatcatt gagaaggaga ctgggaaaca gctggtcatt caggagtcca tcctgatgct
     5521 gcctgaggag gtggaggaag tgatcggcaa caagccagag tctgacatcc tggtgcacac
     5581 cgcctacgac gagtccacag atgagaatgt gatgctgctg acctctgacg cccccgagta
     5641 taagccttgg gccctggtca tccaggattc taacggcgag aataagatca agatgctgag
     5701 cggaggatcc ggaggatctg gaggcagcac caacctgtct gacatcatcg agaaggagac
     5761 aggcaagcag ctggtcatcc aggagagcat cctgatgctg cccgaagaag tcgaagaagt
     5821 gatcggaaac aagcctgaga gcgatatcct ggtccatacc gcctacgacg agagtaccga
     5881 cgaaaatgtg atgctgctga catccgacgc cccagagtat aagccctggg ctctggtcat
     5941 ccaggattcc aacggagaga acaaaatcaa aatgctgtct ggcggctcaa aaagaaccgc
     6001 cgacggcagc gaattcgagc ccaagaagaa gaggaaagtc taaccggtca tcatcaccat
     6061 caccattgag tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     6121 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     6181 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     6241 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     6301 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc gataccgtcg
     6361 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     6421 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctagggtgcc
     6481 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     6541 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     6601 attgggcgca cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     6661 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     6721 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     6781 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     6841 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     6901 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     6961 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     7021 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     7081 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     7141 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     7201 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     7261 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     7321 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     7381 gaagatcctt tgatcttttc tacggggtct gacactcagt ggaacgaaaa ctcacgttaa
     7441 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     7501 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     7561 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     7621 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     7681 atgataccgc gggacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     7741 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     7801 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     7861 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     7921 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     7981 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     8041 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     8101 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     8161 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     8221 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     8281 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     8341 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     8401 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     8461 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     8521 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atcgatctcc cgatccccta
     8581 gggtcgactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     8641 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     8701 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc
     8761 gatgtacggg ccagatat
//
