LOCUS       sequence-386796-        8778 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Mammalian expression of eTd-CBEa.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8778)
  AUTHORS   Chen L, Zhu B, Ru G, Meng H, Yan Y, Hong M, Zhang D, Luan C, Zhang 
            S, Wu H, Gao H, Bai S, Li C, Ding R, Xue N, Lei Z, Chen Y, Guan Y, 
            Siwko S, Cheng Y, Song G, Wang L, Yi C, Liu M, Li D
  TITLE     Re-engineering the adenine deaminase TadA-8e for efficient and 
            specific CRISPR-based cytosine base editing.
  JOURNAL   Nat Biotechnol. 2022 Nov 10. doi: 10.1038/s41587-022-01532-7.
  PUBMED    36357717
REFERENCE   2  (bases 1 to 8778)
  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. 2022 Nov 10. doi: 10.1038/s41587-022-01532-7."
COMMENT     Sequence Label: eTd-CBEa
FEATURES             Location/Qualifiers
     source          1..8778
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        158..361
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     312..332
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     403..422
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        403..421
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             471..491
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(990..1013,1014..1016,1017..5090)
                     /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             990..5090
                     /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             5133..5153
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5163..5219
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             5220..5468
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5499..5747
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5790..5810
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5819..5836
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5859..5876)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5865..6089
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6160..6176)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6160..6176)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6173..6195)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6184..6200
                     /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(6208..6214,6215..6232,6233..6238))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6253..6274
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6391..6408)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6562..7150)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6642..6661)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7321..8112,8113..8181))
                     /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     7944..7963
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8182..8286)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8365..8384)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(8556..8778,1..157)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
       61 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
      121 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
      181 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      241 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
      301 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      361 tggtttagtg aaccgtcaga tccgctagag atccgcggcc gctaatacga ctcactatag
      421 ggagagccgc caccatgaaa cggacagccg acggaagcga gttcgagtca ccaaagaaga
      481 agcggaaagt ctctgaggtg gagttttccc acgagtactg gatgagacat gccctgaccc
      541 tggccaagag ggcacgggat gagagggagg tggcagtggg agccgtgctg gtgctgaaca
      601 atagagtgat cggcgagggc tggctgagag ccatcggcct gcacgaccca acagcccatg
      661 ccgaaattat ggccctgaga cagggcggcc tggtcatgca gaactacaga ctgattgacg
      721 ccaccctgta cgtgacattc gagccttgcg tgatgtgcgc cggcgccatg atccactcta
      781 ggatcggccg cgtggtgttt ggcgtgagga actcaaaaag aggcgccgca ggctccctga
      841 tgaacgtgct gaactacccc ggcatgaatc accgcgtcga aattaccgag ggaatcctgg
      901 cagatgaatg tgccgccctg ctgtgcgatt tctatcggat gcctagacag gtgttcaatg
      961 ctcagaagaa ggcccagagc tccatcaacg acaagaagta cagcatcggc ctggccatcg
     1021 gcaccaactc tgtgggctgg gccgtgatca ccgacgagta caaggtgccc agcaagaaat
     1081 tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa gaacctgatc ggagccctgc
     1141 tgttcgacag cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc agaagaagat
     1201 acaccagacg gaagaaccgg atctgctatc tgcaagagat cttcagcaac gagatggcca
     1261 aggtggacga cagcttcttc cacagactgg aagagtcctt cctggtggaa gaggataaga
     1321 agcacgagcg gcaccccatc ttcggcaaca tcgtggacga ggtggcctac cacgagaagt
     1381 accccaccat ctaccacctg agaaagaaac tggtggacag caccgacaag gccgacctgc
     1441 ggctgatcta tctggccctg gcccacatga tcaagttccg gggccacttc ctgatcgagg
     1501 gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt catccagctg gtgcagacct
     1561 acaaccagct gttcgaggaa aaccccatca acgccagcgg cgtggacgcc aaggccatcc
     1621 tgtctgccag actgagcaag agcagacggc tggaaaatct gatcgcccag ctgcccggcg
     1681 agaagaagaa tggcctgttc ggaaacctga ttgccctgag cctgggcctg acccccaact
     1741 tcaagagcaa cttcgacctg gccgaggatg ccaaactgca gctgagcaag gacacctacg
     1801 acgacgacct ggacaacctg ctggcccaga tcggcgacca gtacgccgac ctgtttctgg
     1861 ccgccaagaa cctgtccgac gccatcctgc tgagcgacat cctgagagtg aacaccgaga
     1921 tcaccaaggc ccccctgagc gcctctatga tcaagagata cgacgagcac caccaggacc
     1981 tgaccctgct gaaagctctc gtgcggcagc agctgcctga gaagtacaaa gagattttct
     2041 tcgaccagag caagaacggc tacgccggct acattgacgg cggagccagc caggaagagt
     2101 tctacaagtt catcaagccc atcctggaaa agatggacgg caccgaggaa ctgctcgtga
     2161 agctgaacag agaggacctg ctgcggaagc agcggacctt cgacaacggc agcatccccc
     2221 accagatcca cctgggagag ctgcacgcca ttctgcggcg gcaggaagat ttttacccat
     2281 tcctgaagga caaccgggaa aagatcgaga agatcctgac cttccgcatc ccctactacg
     2341 tgggccctct ggccagggga aacagcagat tcgcctggat gaccagaaag agcgaggaaa
     2401 ccatcacccc ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc cagagcttca
     2461 tcgagcggat gaccaacttc gataagaacc tgcccaacga gaaggtgctg cccaagcaca
     2521 gcctgctgta cgagtacttc accgtgtata acgagctgac caaagtgaaa tacgtgaccg
     2581 agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc
     2641 tgttcaagac caaccggaaa gtgaccgtga agcagctgaa agaggactac ttcaagaaaa
     2701 tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga tcggttcaac gcctccctgg
     2761 gcacatacca cgatctgctg aaaattatca aggacaagga cttcctggac aatgaggaaa
     2821 acgaggacat tctggaagat atcgtgctga ccctgacact gtttgaggac agagagatga
     2881 tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga caaagtgatg aagcagctga
     2941 agcggcggag atacaccggc tggggcaggc tgagccggaa gctgatcaac ggcatccggg
     3001 acaagcagtc cggcaagaca atcctggatt tcctgaagtc cgacggcttc gccaacagaa
     3061 acttcatgca gctgatccac gacgacagcc tgacctttaa agaggacatc cagaaagccc
     3121 aggtgtccgg ccagggcgat agcctgcacg agcacattgc caatctggcc ggcagccccg
     3181 ccattaagaa gggcatcctg cagacagtga aggtggtgga cgagctcgtg aaagtgatgg
     3241 gccggcacaa gcccgagaac atcgtgatcg aaatggccag agagaaccag accacccaga
     3301 agggacagaa gaacagccgc gagagaatga agcggatcga agagggcatc aaagagctgg
     3361 gcagccagat cctgaaagaa caccccgtgg aaaacaccca gctgcagaac gagaagctgt
     3421 acctgtacta cctgcagaat gggcgggata tgtacgtgga ccaggaactg gacatcaacc
     3481 ggctgtccga ctacgatgtg gaccatatcg tgcctcagag ctttctgaag gacgactcca
     3541 tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg caagagcgac aacgtgccct
     3601 ccgaagaggt cgtgaagaag atgaagaact actggcggca gctgctgaac gccaagctga
     3661 ttacccagag aaagttcgac aatctgacca aggccgagag aggcggcctg agcgaactgg
     3721 ataaggccgg cttcatcaag agacagctgg tggaaacccg gcagatcaca aagcacgtgg
     3781 cacagatcct ggactcccgg atgaacacta agtacgacga gaatgacaag ctgatccggg
     3841 aagtgaaagt gatcaccctg aagtccaagc tggtgtccga tttccggaag gatttccagt
     3901 tttacaaagt gcgcgagatc aacaactacc accacgccca cgacgcctac ctgaacgccg
     3961 tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga aagcgagttc gtgtacggcg
     4021 actacaaggt gtacgacgtg cggaagatga tcgccaagag cgagcaggaa atcggcaagg
     4081 ctaccgccaa gtacttcttc tacagcaaca tcatgaactt tttcaagacc gagattaccc
     4141 tggccaacgg cgagatccgg aagcggcctc tgatcgagac aaacggcgaa accggggaga
     4201 tcgtgtggga taagggccgg gattttgcca ccgtgcggaa agtgctgagc atgccccaag
     4261 tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt cagcaaagag tctatcctgc
     4321 ccaagaggaa cagcgataag ctgatcgcca gaaagaagga ctgggaccct aagaagtacg
     4381 gcggcttcga cagccccacc gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg
     4441 gcaagtccaa gaaactgaag agtgtgaaag agctgctggg gatcaccatc atggaaagaa
     4501 gcagcttcga gaagaatccc atcgactttc tggaagccaa gggctacaaa gaagtgaaaa
     4561 aggacctgat catcaagctg cctaagtact ccctgttcga gctggaaaac ggccggaaga
     4621 gaatgctggc ctctgccggc gaactgcaga agggaaacga actggccctg ccctccaaat
     4681 atgtgaactt cctgtacctg gccagccact atgagaagct gaagggctcc cccgaggata
     4741 atgagcagaa acagctgttt gtggaacagc acaagcacta cctggacgag atcatcgagc
     4801 agatcagcga gttctccaag agagtgatcc tggccgacgc taatctggac aaagtgctgt
     4861 ccgcctacaa caagcaccgg gataagccca tcagagagca ggccgagaat atcatccacc
     4921 tgtttaccct gaccaatctg ggagcccctg ccgccttcaa gtactttgac accaccatcg
     4981 accggaagag gtacaccagc accaaagagg tgctggacgc caccctgatc caccagagca
     5041 tcaccggcct gtacgagaca cggatcgacc tgtctcagct gggaggtgac tctggcggct
     5101 caaaaagaac cgccgacggc agcgaattcg agcccaagaa gaagaggaaa gtcggaagcg
     5161 gagctactaa cttcagcctg ctgaagcagg ctggagacgt ggaggagaac cctggaccta
     5221 ctaatctgag cgacatcatt gagaaggaga ctgggaaaca gctggtcatt caggagtcca
     5281 tcctgatgct gcctgaggag gtggaggaag tgatcggcaa caagccagag tctgacatcc
     5341 tggtgcacac cgcctacgac gagtccacag atgagaatgt gatgctgctg acctctgacg
     5401 cccccgagta taagccttgg gccctggtca tccaggattc taacggcgag aataagatca
     5461 agatgctgag cggaggatcc ggaggatctg gaggcagcac caacctgtct gacatcatcg
     5521 agaaggagac aggcaagcag ctggtcatcc aggagagcat cctgatgctg cccgaagaag
     5581 tcgaagaagt gatcggaaac aagcctgaga gcgatatcct ggtccatacc gcctacgacg
     5641 agagtaccga cgaaaatgtg atgctgctga catccgacgc cccagagtat aagccctggg
     5701 ctctggtcat ccaggattcc aacggagaga acaaaatcaa aatgctgtct ggcggctcaa
     5761 aaagaaccgc cgacggcagc gaattcgagc ccaagaagaa gaggaaagtc taaccggtca
     5821 tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc cttctagttg
     5881 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     5941 cactgttctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     6001 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     6061 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     6121 gataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     6181 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     6241 ctagggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     6301 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     6361 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     6421 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     6481 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6541 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6601 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6661 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6721 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     6781 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6841 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6901 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6961 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     7021 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     7081 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     7141 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacactcagt ggaacgaaaa
     7201 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     7261 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     7321 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     7381 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     7441 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     7501 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     7561 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     7621 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7681 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7741 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7801 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7861 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     7921 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7981 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     8041 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     8101 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     8161 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     8221 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     8281 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atcgatctcc
     8341 cgatccccta gggtcgactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     8401 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
     8461 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc
     8521 gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta
     8581 atagtaatca attacggggt cattagttca tagcccatat attgagttcc gcgttacata
     8641 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     8701 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     8761 gtatttacgg taaactgc
//
