LOCUS       sequence-238826-        8355 bp    DNA     circular SYN 14-APR-2025
DEFINITION  For in vitro transcription to produce Cas9 mRNA for microinjection 
            into mouse embryos.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8355)
  AUTHORS   Gu B, Posfai E, Rossant J
  TITLE     Efficient generation of targeted large insertions by microinjection 
            into two-cell-stage mouse embryos.
  JOURNAL   Nat Biotechnol. 2018 Jun 11. pii: nbt.4166. doi: 10.1038/nbt.4166.
  PUBMED    29889212
REFERENCE   2  (bases 1 to 8355)
  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. 2018 Jun 11. pii: nbt.4166. doi: 10.1038/nbt.4166."
COMMENT     Sequence Label: PCS2+ Cas9
FEATURES             Location/Qualifiers
     source          1..8355
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..66
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             73..93
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             118..4218
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             118..4218
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4219..4266
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     polyA_signal    4320..4454
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4344..4363)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4398..4417
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4567..4587)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4567..4585)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4606..4622)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4606..4622)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4619..4641)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4630..4646
                     /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(4654..4660,4661..4678,4679..4684))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4699..4720
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4837..4854)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5008..5596)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5088..5107)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5767..6558,6559..6627))
                     /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     6390..6409
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6628..6732)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6758..7213)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6895..6916)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7107..7126
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        7284..8268
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        8303..8321
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     8303..8320
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gactataagg accacgacgg agactacaag gatcatgata ttgattacaa agacgatgac
       61 gataagatgg ccccaaagaa gaagcggaag gtcggtatcc acggagtccc agcagccgac
      121 aagaagtaca gcatcggcct ggacatcggc accaactctg tgggctgggc cgtgatcacc
      181 gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga ccggcacagc
      241 atcaagaaga acctgatcgg agccctgctg ttcgacagcg gcgaaacagc cgaggccacc
      301 cggctgaaga gaaccgccag aagaagatac accagacgga agaaccggat ctgctatctg
      361 caagagatct tcagcaacga gatggccaag gtggacgaca gcttcttcca cagactggaa
      421 gagtccttcc tggtggaaga ggataagaag cacgagcggc accccatctt cggcaacatc
      481 gtggacgagg tggcctacca cgagaagtac cccaccatct accacctgag aaagaaactg
      541 gtggacagca ccgacaaggc cgacctgcgg ctgatctatc tggccctggc ccacatgatc
      601 aagttccggg gccacttcct gatcgagggc gacctgaacc ccgacaacag cgacgtggac
      661 aagctgttca tccagctggt gcagacctac aaccagctgt tcgaggaaaa ccccatcaac
      721 gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg
      781 gaaaatctga tcgcccagct gcccggcgag aagaagaatg gcctgttcgg aaacctgatt
      841 gccctgagcc tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc
      901 aaactgcagc tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc
      961 ggcgaccagt acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg
     1021 agcgacatcc tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc
     1081 aagagatacg acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag
     1141 ctgcctgaga agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac
     1201 attgacggcg gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag
     1261 atggacggca ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag
     1321 cggaccttcg acaacggcag catcccccac cagatccacc tgggagagct gcacgccatt
     1381 ctgcggcggc aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag
     1441 atcctgacct tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc
     1501 gcctggatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg
     1561 gacaagggcg cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg
     1621 cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac
     1681 gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc
     1741 gagcagaaaa aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag
     1801 cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc
     1861 gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag
     1921 gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc
     1981 ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg
     2041 ttcgacgaca aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg
     2101 agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc
     2161 ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg
     2221 acctttaaag aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag
     2281 cacattgcca atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag
     2341 gtggtggacg agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa
     2401 atggccagag agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag
     2461 cggatcgaag agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa
     2521 aacacccagc tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg
     2581 tacgtggacc aggaactgga catcaaccgg ctgtccgact acgatgtgga ccatatcgtg
     2641 cctcagagct ttctgaagga cgactccatc gacaacaagg tgctgaccag aagcgacaag
     2701 aaccggggca agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac
     2761 tggcggcagc tgctgaacgc caagctgatt acccagagaa agttcgacaa tctgaccaag
     2821 gccgagagag gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg
     2881 gaaacccggc agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag
     2941 tacgacgaga atgacaagct gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg
     3001 gtgtccgatt tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac
     3061 cacgcccacg acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct
     3121 aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc
     3181 gccaagagcg agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc
     3241 atgaactttt tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg
     3301 atcgagacaa acggcgaaac cggggagatc gtgtgggata agggccggga ttttgccacc
     3361 gtgcggaaag tgctgagcat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca
     3421 ggcggcttca gcaaagagtc tatcctgccc aagaggaaca gcgataagct gatcgccaga
     3481 aagaaggact gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct
     3541 gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag
     3601 ctgctgggga tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg
     3661 gaagccaagg gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc
     3721 ctgttcgagc tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag
     3781 ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat
     3841 gagaagctga agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac
     3901 aagcactacc tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg
     3961 gccgacgcta atctggacaa agtgctgtcc gcctacaaca agcaccggga taagcccatc
     4021 agagagcagg ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc
     4081 gccttcaagt actttgacac caccatcgac cggaagaggt acaccagcac caaagaggtg
     4141 ctggacgcca ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg
     4201 tctcagctgg gaggcgacaa aaggccggcg gccacgaaaa aggccggcca ggcaaaaaag
     4261 aaaaagtaag aattcaaggc ctctcgagcc tctagaacta tagtgagtcg tattacgtag
     4321 atccagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     4381 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     4441 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
     4501 tgtgggaggt tttttaattc gcggccgcgg cgccaatgca ttgggcccgg tacccagctt
     4561 ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc
     4621 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggga gcataaagtg
     4681 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     4741 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     4801 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4861 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4921 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4981 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5041 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5101 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5161 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5221 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5281 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5341 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5401 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     5461 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5521 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5581 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5641 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5701 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5761 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5821 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5881 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5941 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6001 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6061 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6121 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6181 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6241 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6301 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6361 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6421 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6481 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6541 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6601 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6661 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6721 aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg
     6781 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     6841 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     6901 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     6961 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     7021 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     7081 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     7141 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     7201 ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     7261 cgggcctctt cgctattacg ccagtcgacc gccaattcaa tatggcgtat atggactcat
     7321 gccaattcaa tatggtggat ctggacctgt gccaattcaa tatggcgtat atggactcgt
     7381 gccaattcaa tatggtggat ctggacccca gccaattcaa tatggcggac ttggcaccat
     7441 gccaattcaa tatggcggac ctggcactgt gccaactggg gaggggtcta cttggcacgg
     7501 tgccaagttt gaggaggggt cttggccctg tgccaagtcc gccatattga attggcatgg
     7561 tgccaataat ggcggccata ttggctatat gccaggatca atatataggc aatatccaat
     7621 atggccctat gccaatatgg ctattggcca ggttcaatac tatgtattgg ccctatgcca
     7681 tatagtattc catatatggg ttttcctatt gacgtagata gcccctccca atgggcggtc
     7741 ccatatacca tatatggggc ttcctaatac cgcccatagc cactccccca ttgacgtcaa
     7801 tggtctctat atatggtctt tcctattgac gtcatatggg cggtcctatt gacgtatatg
     7861 gcgcctcccc cattgacgtc aattacggta aatggcccgc ctggctcaat gcccattgac
     7921 gtcaatagga ccacccacca ttgacgtcaa tgggatggct cattgcccat tcatatccgt
     7981 tctcacgccc cctattgacg tcaatgacgg taaatggccc acttggcagt acatcaatat
     8041 ctattaatag taacttggca agtacattac tattggaagt acgccagggt acattggcag
     8101 tactcccatt gacgtcaatg gcggtaaatg gcccgcgatg gctgccaagt acatccccat
     8161 tgacgtcaat ggggaggggc aatgacgcaa atgggcgttc cattgacgta aatgggcggt
     8221 aggcgtgcct aatgggaggt ctatataagc aatgctcgtt tagggaaccg ccattctgcc
     8281 tggggacgtc ggagcaagct tgatttaggt gacactatag aatacaagct acttgttctt
     8341 tttgcaggat ccatg
//
