LOCUS       sequence-244237-        8712 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the Rad51(F86E)–Cas9(D10A) fusion construct in mammalian 
            cells under CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8712)
  AUTHORS   Rees HA, Yeh WH, Liu DR
  TITLE     Development of hRad51-Cas9 nickase fusions that mediate HDR without 
            double-stranded breaks.
  JOURNAL   Nat Commun. 2019 May 17;10(1):2212. doi: 10.1038/s41467-019-09983-4.
  PUBMED    31101808
REFERENCE   2  (bases 1 to 8712)
  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: "2019-05-17"; volume: "10(1):2212. doi"; pages: " 
            10.1038/s41467-019-09983-4"
COMMENT     Sequence Label: hRad51(F86E)–Cas9(D10A)
FEATURES             Location/Qualifiers
     source          1..8712
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        132..335
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     286..306
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     377..396
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        377..395
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      403..412
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             415..435
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1570..1593,1594..1596,1597..5670)
                     /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             1570..5670
                     /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             5671..5718
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5727..5744
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5767..5784)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5773..5997
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6068..6084)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6068..6084)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6081..6103)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6092..6108
                     /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(6116..6122,6123..6140,6141..6146))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6161..6182
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6299..6316)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6470..7058)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6550..6569)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7229..8020,8021..8089))
                     /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     7852..7871
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8090..8194)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8273..8292)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(8464..8712,1..131)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
       61 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      121 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      181 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      241 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      301 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      361 agagatccgc ggccgctaat acgactcact atagggagag ccgccaccat ggccccaaag
      421 aagaagcgga aggtcggtat ccacggagtc ccagcagccg cgatgcagat gcagttggaa
      481 gcgaatgcag atactagtgt cgaggaagag tcatttggcc cgcaacccat ctcgcgttta
      541 gagcaatgtg gcatcaatgc aaacgatgtg aaaaaattag aggaagctgg attccacacg
      601 gtcgaagcgg tcgcatacgc accgaaaaaa gagctgatca acatcaaagg catcagcgag
      661 gcgaaagccg ataagattct tgcagaggcg gcgaaattag ttcccatggg agaaacgacg
      721 gcgactgagt tccatcaacg tcgttccgag atcattcaaa tcacgaccgg aagcaaggag
      781 ttggataaac tgcttcaagg aggcatcgaa actggctcga tcactgaaat gtttggagag
      841 tttcgtactg gcaaaaccca gatctgccac acattggccg tcacttgcca gcttccgatc
      901 gatcgtggtg ggggcgaggg gaaggccatg tacattgata ctgaggggac gtttcgtccc
      961 gaacgccttc ttgcggtggc agagcgttac ggcttgtcag ggtctgatgt tcttgacaat
     1021 gttgcctatg ctcgcgcatt taataccgac caccagacgc agcttctgta ccaagcaagc
     1081 gcgatgatgg tagagtctcg ttacgcttta ttgattgttg actcagcaac agcgttgtac
     1141 cgcacagatt attctgggcg cggcgaattg tcggcacgcc aaatgcattt agcccgcttt
     1201 ttgcgtatgt tgctgcgcct tgcagacgaa tttggggtgg ctgtggtgat caccaaccaa
     1261 gttgtagcac aggtggacgg tgccgctatg tttgccgctg atccgaaaaa acccatcgga
     1321 gggaatatta tcgcacatgc atcgacaact cgcttatatc tgcgtaaggg ccgtggagaa
     1381 acacgcatct gcaaaatcta tgactcccca tgcctgccgg aggcggaagc catgttcgca
     1441 attaacgcag atggcgtagg ggacgccaaa gattctggtg gttcttctgg tggttctagc
     1501 ggcagcgaga ctcccgggac ctcagagtcc gccacacccg aaagttctgg tggttcttct
     1561 ggtggttctg acaagaagta cagcatcggc ctggccatcg gcaccaactc tgtgggctgg
     1621 gccgtgatca ccgacgagta caaggtgccc agcaagaaat tcaaggtgct gggcaacacc
     1681 gaccggcaca gcatcaagaa gaacctgatc ggagccctgc tgttcgacag cggcgaaaca
     1741 gccgaggcca cccggctgaa gagaaccgcc agaagaagat acaccagacg gaagaaccgg
     1801 atctgctatc tgcaagagat cttcagcaac gagatggcca aggtggacga cagcttcttc
     1861 cacagactgg aagagtcctt cctggtggaa gaggataaga agcacgagcg gcaccccatc
     1921 ttcggcaaca tcgtggacga ggtggcctac cacgagaagt accccaccat ctaccacctg
     1981 agaaagaaac tggtggacag caccgacaag gccgacctgc ggctgatcta tctggccctg
     2041 gcccacatga tcaagttccg gggccacttc ctgatcgagg gcgacctgaa ccccgacaac
     2101 agcgacgtgg acaagctgtt catccagctg gtgcagacct acaaccagct gttcgaggaa
     2161 aaccccatca acgccagcgg cgtggacgcc aaggccatcc tgtctgccag actgagcaag
     2221 agcagacggc tggaaaatct gatcgcccag ctgcccggcg agaagaagaa tggcctgttc
     2281 ggaaacctga ttgccctgag cctgggcctg acccccaact tcaagagcaa cttcgacctg
     2341 gccgaggatg ccaaactgca gctgagcaag gacacctacg acgacgacct ggacaacctg
     2401 ctggcccaga tcggcgacca gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac
     2461 gccatcctgc tgagcgacat cctgagagtg aacaccgaga tcaccaaggc ccccctgagc
     2521 gcctctatga tcaagagata cgacgagcac caccaggacc tgaccctgct gaaagctctc
     2581 gtgcggcagc agctgcctga gaagtacaaa gagattttct tcgaccagag caagaacggc
     2641 tacgccggct acattgacgg cggagccagc caggaagagt tctacaagtt catcaagccc
     2701 atcctggaaa agatggacgg caccgaggaa ctgctcgtga agctgaacag agaggacctg
     2761 ctgcggaagc agcggacctt cgacaacggc agcatccccc accagatcca cctgggagag
     2821 ctgcacgcca ttctgcggcg gcaggaagat ttttacccat tcctgaagga caaccgggaa
     2881 aagatcgaga agatcctgac cttccgcata ccctactacg tgggccctct ggccagggga
     2941 aacagcagat tcgcctggat gaccagaaag agcgaggaaa ccatcacccc ctggaacttc
     3001 gaggaagtgg tggacaaggg cgcttccgcc cagagcttca tcgagcggat gaccaacttc
     3061 gataagaacc tgcccaacga gaaggtgctg cccaagcaca gcctgctgta cgagtacttc
     3121 accgtgtata acgagctgac caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc
     3181 ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc tgttcaagac caaccggaaa
     3241 gtgaccgtga agcagctgaa agaggactac ttcaagaaaa tcgagtgctt cgactccgtg
     3301 gaaatctccg gcgtggaaga tcggttcaac gcctccctgg gcacatacca cgatctgctg
     3361 aaaattatca aggacaagga cttcctggac aatgaggaaa acgaggacat tctggaagat
     3421 atcgtgctga ccctgacact gtttgaggac agagagatga tcgaggaacg gctgaaaacc
     3481 tatgcccacc tgttcgacga caaagtgatg aagcagctga agcggcggag atacaccggc
     3541 tggggcaggc tgagccggaa gctgatcaac ggcatccggg acaagcagtc cggcaagaca
     3601 atcctggatt tcctgaagtc cgacggcttc gccaacagaa acttcatgca gctgatccac
     3661 gacgacagcc tgacctttaa agaggacatc cagaaagccc aggtgtccgg ccagggcgat
     3721 agcctgcacg agcacattgc caatctggcc ggcagccccg ccattaagaa gggcatcctg
     3781 cagacagtga aggtggtgga cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac
     3841 atcgtgatcg aaatggccag agagaaccag accacccaga agggacagaa gaacagccgc
     3901 gagagaatga agcggatcga agagggcatc aaagagctgg gcagccagat cctgaaagaa
     3961 caccccgtgg aaaacaccca gctgcagaac gagaagctgt acctgtacta cctgcagaat
     4021 gggcgggata tgtacgtgga ccaggaactg gacatcaacc ggctgtccga ctacgatgtg
     4081 gaccatatcg tgcctcagag ctttctgaag gacgactcca tcgacaacaa ggtgctgacc
     4141 agaagcgaca agaaccgggg caagagcgac aacgtgccct ccgaagaggt cgtgaagaag
     4201 atgaagaact actggcggca gctgctgaac gccaagctga ttacccagag aaagttcgac
     4261 aatctgacca aggccgagag aggcggcctg agcgaactgg ataaggccgg cttcatcaag
     4321 agacagctgg tggaaacccg gcagatcaca aagcacgtgg cacagatcct ggactcccgg
     4381 atgaacacta agtacgacga gaatgacaag ctgatccggg aagtgaaagt gatcaccctg
     4441 aagtccaagc tggtgtccga tttccggaag gatttccagt tttacaaagt gcgcgagatc
     4501 aacaactacc accacgccca cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc
     4561 aaaaagtacc ctaagctgga aagcgagttc gtgtacggcg actacaaggt gtacgacgtg
     4621 cggaagatga tcgccaagag cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc
     4681 tacagcaaca tcatgaactt tttcaagacc gagattaccc tggccaacgg cgagatccgg
     4741 aagcggcctc tgatcgagac aaacggcgaa accggggaga tcgtgtggga taagggccgg
     4801 gattttgcca ccgtgcggaa agtgctgagc atgccccaag tgaatatcgt gaaaaagacc
     4861 gaggtgcaga caggcggctt cagcaaagag tctatcctgc ccaagaggaa cagcgataag
     4921 ctgatcgcca gaaagaagga ctgggaccct aagaagtacg gcggcttcga cagccccacc
     4981 gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag
     5041 agtgtgaaag agctgctggg gatcaccatc atggaaagaa gcagcttcga gaagaatccc
     5101 atcgactttc tggaagccaa gggctacaaa gaagtgaaaa aggacctgat catcaagctg
     5161 cctaagtact ccctgttcga gctggaaaac ggccggaaga gaatgctggc ctctgccggc
     5221 gaactgcaga agggaaacga actggccctg ccctccaaat atgtgaactt cctgtacctg
     5281 gccagccact atgagaagct gaagggctcc cccgaggata atgagcagaa acagctgttt
     5341 gtggaacagc acaagcacta cctggacgag atcatcgagc agatcagcga gttctccaag
     5401 agagtgatcc tggccgacgc taatctggac aaagtgctgt ccgcctacaa caagcaccgg
     5461 gataagccca tcagagagca ggccgagaat atcatccacc tgtttaccct gaccaatctg
     5521 ggagcccctg ccgccttcaa gtactttgac accaccatcg accggaagag gtacaccagc
     5581 accaaagagg tgctggacgc caccctgatc caccagagca tcaccggcct gtacgagaca
     5641 cggatcgacc tgtctcagct gggaggcgac aaaaggccgg cggccacgaa aaaggccggc
     5701 caggcaaaaa agaaaaagtg accggtcatc atcaccatca ccattgagtt taaacccgct
     5761 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     5821 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     5881 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     5941 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     6001 ctgaggcgga aagaaccagc tggggctcga taccgtcgac ctctagctag agcttggcgt
     6061 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     6121 tacgagccgg aagcataaag tgtaaagcct agggtgccta atgagtgagc taactcacat
     6181 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     6241 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     6301 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     6361 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     6421 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6481 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6541 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6601 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6661 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6721 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6781 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6841 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6901 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6961 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     7021 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     7081 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     7141 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     7201 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     7261 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     7321 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     7381 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     7441 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     7501 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     7561 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     7621 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     7681 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     7741 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     7801 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     7861 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     7921 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7981 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     8041 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     8101 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     8161 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     8221 acgtcgacgg atcgggagat cgatctcccg atcccctagg gtcgactctc agtacaatct
     8281 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     8341 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     8401 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
     8461 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
     8521 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     8581 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     8641 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     8701 atcaagtgta tc
//
