LOCUS       sequence-424801-        8857 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template for in vitro transcription of CBE6a (with SpCas9).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8857)
  AUTHORS   Zhang E, Neugebauer ME, Krasnow NA, Liu DR
  TITLE     Phage-assisted evolution of highly active cytosine base editors with
            enhanced selectivity and minimal sequence context preference.
  JOURNAL   Nat Commun. 2024 Feb 24;15(1):1697. doi: 10.1038/s41467-024-45969-7.
  PUBMED    38402281
REFERENCE   2  (bases 1 to 8857)
  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 
            Commun."; date: "2024-02-24"; volume: "15(1):1697. doi"; pages: " 
            10.1038/s41467-024-45969-7"
COMMENT     Sequence Label: SpCas9-CBE6a-IVT
FEATURES             Location/Qualifiers
     source          1..8857
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             720..740
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1338..1361,1362..1364,1365..5438)
                     /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             1338..5438
                     /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             5469..5717
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5748..5996
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6039..6059
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    6167..6391
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6462..6478)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6462..6478)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6475..6497)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6486..6502
                     /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(6510..6516,6517..6534,6535..6540))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6555..6576
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6693..6710)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6864..7452)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6944..6963)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7623..8414,8415..8483))
                     /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     8246..8265
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8484..8588)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8667..8686)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
      601 agatctcgag ctcggtacct aatacgacac actataagga aataagagag aaaagaagag
      661 taagaagaaa tataagagcc accatgaaac ggacagccga cggaagcgag ttcgagtcac
      721 caaagaagaa gcggaaagtc agttctgagg tggagttttc ccacgagtac tggatgagac
      781 atgccctgac cctggccaag agggcacggg atgagggaga ggcgcctgtg ggagccgtgc
      841 tggtgctgaa caatagagtg atcggcgagg gctggatcag acgtatcggc ctgcacgacc
      901 caacagccca tgccgaaatt atggccctga gacagggcgg cctggtcatg cagaacccca
      961 gactgattga cgccaccctg tacgtgacat tcgagccttg cgtgatgtgc gccggcgcca
     1021 tgatcaactc taggatcggc cgcgtggtgt ttggcgtgag gaactcaaaa agaggcgccg
     1081 caggctccct gatgaacgtg ctgaactacc ccggcatgaa tcaccgcgtc gaaattaccg
     1141 agggaatcct ggcagatgaa tgtgccgccc tgctgtgcga tttctatcgg atgcctagac
     1201 aggtgttcaa tgctcagaag aaggcccaga gctccatcaa ctctggcgga tctagcggag
     1261 gatcctctgg cagcgagaca ccaggaacaa gcgagtcagc aacaccagag agcagtggcg
     1321 gcagcagcgg cggcagcgac aagaagtaca gcatcggcct ggccatcggc accaactctg
     1381 tgggctgggc cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg
     1441 gcaacaccga ccggcacagc atcaagaaga acctgatcgg agccctgctg ttcgacagcg
     1501 gcgaaacagc cgaggccacc cggctgaaga gaaccgccag aagaagatac accagacgga
     1561 agaaccggat ctgctatctg caagagatct tcagcaacga gatggccaag gtggacgaca
     1621 gcttcttcca cagactggaa gagtccttcc tggtggaaga ggataagaag cacgagcggc
     1681 accccatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac cccaccatct
     1741 accacctgag aaagaaactg gtggacagca ccgacaaggc cgacctgcgg ctgatctatc
     1801 tggccctggc ccacatgatc aagttccggg gccacttcct gatcgagggc gacctgaacc
     1861 ccgacaacag cgacgtggac aagctgttca tccagctggt gcagacctac aaccagctgt
     1921 tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa ggccatcctg tctgccagac
     1981 tgagcaagag cagacggctg gaaaatctga tcgcccagct gcccggcgag aagaagaatg
     2041 gcctgttcgg aaacctgatt gccctgagcc tgggcctgac ccccaacttc aagagcaact
     2101 tcgacctggc cgaggatgcc aaactgcagc tgagcaagga cacctacgac gacgacctgg
     2161 acaacctgct ggcccagatc ggcgaccagt acgccgacct gtttctggcc gccaagaacc
     2221 tgtccgacgc catcctgctg agcgacatcc tgagagtgaa caccgagatc accaaggccc
     2281 ccctgagcgc ctctatgatc aagagatacg acgagcacca ccaggacctg accctgctga
     2341 aagctctcgt gcggcagcag ctgcctgaga agtacaaaga gattttcttc gaccagagca
     2401 agaacggcta cgccggctac attgacggcg gagccagcca ggaagagttc tacaagttca
     2461 tcaagcccat cctggaaaag atggacggca ccgaggaact gctcgtgaag ctgaacagag
     2521 aggacctgct gcggaagcag cggaccttcg acaacggcag catcccccac cagatccacc
     2581 tgggagagct gcacgccatt ctgcggcggc aggaagattt ttacccattc ctgaaggaca
     2641 accgggaaaa gatcgagaag atcctgacct tccgcatccc ctactacgtg ggccctctgg
     2701 ccaggggaaa cagcagattc gcctggatga ccagaaagag cgaggaaacc atcaccccct
     2761 ggaacttcga ggaagtggtg gacaagggcg cttccgccca gagcttcatc gagcggatga
     2821 ccaacttcga taagaacctg cccaacgaga aggtgctgcc caagcacagc ctgctgtacg
     2881 agtacttcac cgtgtataac gagctgacca aagtgaaata cgtgaccgag ggaatgagaa
     2941 agcccgcctt cctgagcggc gagcagaaaa aggccatcgt ggacctgctg ttcaagacca
     3001 accggaaagt gaccgtgaag cagctgaaag aggactactt caagaaaatc gagtgcttcg
     3061 actccgtgga aatctccggc gtggaagatc ggttcaacgc ctccctgggc acataccacg
     3121 atctgctgaa aattatcaag gacaaggact tcctggacaa tgaggaaaac gaggacattc
     3181 tggaagatat cgtgctgacc ctgacactgt ttgaggacag agagatgatc gaggaacggc
     3241 tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag cggcggagat
     3301 acaccggctg gggcaggctg agccggaagc tgatcaacgg catccgggac aagcagtccg
     3361 gcaagacaat cctggatttc ctgaagtccg acggcttcgc caacagaaac ttcatgcagc
     3421 tgatccacga cgacagcctg acctttaaag aggacatcca gaaagcccag gtgtccggcc
     3481 agggcgatag cctgcacgag cacattgcca atctggccgg cagccccgcc attaagaagg
     3541 gcatcctgca gacagtgaag gtggtggacg agctcgtgaa agtgatgggc cggcacaagc
     3601 ccgagaacat cgtgatcgaa atggccagag agaaccagac cacccagaag ggacagaaga
     3661 acagccgcga gagaatgaag cggatcgaag agggcatcaa agagctgggc agccagatcc
     3721 tgaaagaaca ccccgtggaa aacacccagc tgcagaacga gaagctgtac ctgtactacc
     3781 tgcagaatgg gcgggatatg tacgtggacc aggaactgga catcaaccgg ctgtccgact
     3841 acgatgtgga ccatatcgtg cctcagagct ttctgaagga cgactccatc gacaacaagg
     3901 tgctgaccag aagcgacaag aaccggggca agagcgacaa cgtgccctcc gaagaggtcg
     3961 tgaagaagat gaagaactac tggcggcagc tgctgaacgc caagctgatt acccagagaa
     4021 agttcgacaa tctgaccaag gccgagagag gcggcctgag cgaactggat aaggccggct
     4081 tcatcaagag acagctggtg gaaacccggc agatcacaaa gcacgtggca cagatcctgg
     4141 actcccggat gaacactaag tacgacgaga atgacaagct gatccgggaa gtgaaagtga
     4201 tcaccctgaa gtccaagctg gtgtccgatt tccggaagga tttccagttt tacaaagtgc
     4261 gcgagatcaa caactaccac cacgcccacg acgcctacct gaacgccgtc gtgggaaccg
     4321 ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt
     4381 acgacgtgcg gaagatgatc gccaagagcg agcaggaaat cggcaaggct accgccaagt
     4441 acttcttcta cagcaacatc atgaactttt tcaagaccga gattaccctg gccaacggcg
     4501 agatccggaa gcggcctctg atcgagacaa acggcgaaac cggggagatc gtgtgggata
     4561 agggccggga ttttgccacc gtgcggaaag tgctgagcat gccccaagtg aatatcgtga
     4621 aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc aagaggaaca
     4681 gcgataagct gatcgccaga aagaaggact gggaccctaa gaagtacggc ggcttcgaca
     4741 gccccaccgt ggcctattct gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga
     4801 aactgaagag tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc agcttcgaga
     4861 agaatcccat cgactttctg gaagccaagg gctacaaaga agtgaaaaag gacctgatca
     4921 tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg ccggaagaga atgctggcct
     4981 ctgccggcga actgcagaag ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc
     5041 tgtacctggc cagccactat gagaagctga agggctcccc cgaggataat gagcagaaac
     5101 agctgtttgt ggaacagcac aagcactacc tggacgagat catcgagcag atcagcgagt
     5161 tctccaagag agtgatcctg gccgacgcta atctggacaa agtgctgtcc gcctacaaca
     5221 agcaccggga taagcccatc agagagcagg ccgagaatat catccacctg tttaccctga
     5281 ccaatctggg agcccctgcc gccttcaagt actttgacac caccatcgac cggaagaggt
     5341 acaccagcac caaagaggtg ctggacgcca ccctgatcca ccagagcatc accggcctgt
     5401 acgagacacg gatcgacctg tctcagctgg gaggtgacag cggcgggagc ggcgggagcg
     5461 gggggagcac taatctgagc gacatcattg agaaggagac tgggaaacag ctggtcattc
     5521 aggagtccat cctgatgctg cctgaggagg tggaggaagt gatcggcaac aagccagagt
     5581 ctgacatcct ggtgcacacc gcctacgacg agtccacaga tgagaatgtg atgctgctga
     5641 cctctgacgc ccccgagtat aagccttggg ccctggtcat ccaggattct aacggcgaga
     5701 ataagatcaa gatgctgagc ggaggatccg gaggatctgg aggcagcacc aacctgtctg
     5761 acatcatcga gaaggagaca ggcaagcagc tggtcatcca ggagagcatc ctgatgctgc
     5821 ccgaagaagt cgaagaagtg atcggaaaca agcctgagag cgatatcctg gtccataccg
     5881 cctacgacga gagtaccgac gaaaatgtga tgctgctgac atccgacgcc ccagagtata
     5941 agccctgggc tctggtcatc caggattcca acggagagaa caaaatcaaa atgctgtctg
     6001 gcggctcaaa aagaaccgcc gacggcagcg aattcgagcc caagaagaag aggaaagtct
     6061 aattaattaa gctgccttct gcggggcttg ccttctggcc atgcccttct tctctccctt
     6121 gcacctgtac ctcttggtct ttgaataaag cctgagtagg aagcgactgt gccttctagt
     6181 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     6241 cccactgtcc tttcctaata aaatgagaaa attgcatcgc attgtctgag taggtgtcat
     6301 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     6361 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     6421 tcgataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     6481 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     6541 gcctagggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     6601 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     6661 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     6721 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     6781 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6841 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6901 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6961 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     7021 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     7081 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     7141 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     7201 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     7261 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     7321 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     7381 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     7441 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacactca gtggaacgaa
     7501 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     7561 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     7621 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     7681 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     7741 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     7801 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     7861 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     7921 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     7981 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     8041 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     8101 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     8161 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     8221 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     8281 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     8341 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     8401 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     8461 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     8521 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     8581 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatcgatct
     8641 cccgatcccc tagggtctac tctcagtaca atctgctctg atgccgcata gttaagccag
     8701 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
     8761 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt
     8821 gcgctgcttc gcgatgtacg ggccagatat acgcgtt
//
