LOCUS       sequence-199211-        9810 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cas9 Homologous Recombination Reporter. SpCas9D10A nickase and a 
            pair of sgRNA targeting hLMNA. Generates breaks with 44bp 5' 
            overhangs. TagBFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9810)
  AUTHORS   Arnoult N, Correia A, Ma J, Merlo A, Garcia-Gomez S, Maric M, 
            Tognetti M, Benner CW, Boulton SJ, Saghatelian A, Karlseder J
  TITLE     Regulation of DNA repair pathway choice in S and G2 phases by the 
            NHEJ inhibitor CYREN.
  JOURNAL   Nature. 2017 Sep 20;549(7673):548-552. doi: 10.1038/nature24023.
  PUBMED    28959974
REFERENCE   2  (bases 1 to 9810)
  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"; doi: "10"; journalName: 
            "Nature"; date: "2017-09-20- 20"; volume: "549"; issue: "7673"; 
            pages: "548-552"
COMMENT     Sequence Label: pUC CBA-SpCas9D10Anickase.EF1a-BFP.sgLMNApair1
FEATURES             Location/Qualifiers
     source          1..9810
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1..21
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     172..191
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        271..346
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        396..607
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             670..1371
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     promoter        1393..1607
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     1528..1547
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     misc_RNA        1637..1712
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        1745..2030
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        2032..2309
                     /label=chicken beta-actin promoter
     intron          2310..2537
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      2549..2558
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2558..2623
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2630..2650
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2675..2698,2699..2701,2702..6775)
                     /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             2675..6775
                     /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             6776..6823
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(6851..6868)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6857..7064
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   7073..7213
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   7073..7202
                     /label=AAV2 ITR
     rep_origin      7288..7743
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7375..7394)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7585..7606
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7760..7779)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7879..7901
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7939..7957)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8025..8129
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8130..8198,8199..8990)
                     /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     complement(8348..8367)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9161..9749
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9650..9669
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg ggttggcagc gctgcccgcg gttttagagc tagaaatagc aagttaaaat
      301 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttagcgcgtg
      361 cgccaattct gcagacatga atcaacctct ggattgggca gagcgcacat cgcccacagt
      421 ccccgagaag ttggggggag gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg
      481 ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga
      541 accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag
      601 aacacagctc tggctaacta gagaacccac tgtagtgaac cgtcagatcg cctggagacg
      661 ccatccacga tgtccgagct gattaaggag aacatgcaca tgaagctgta catggagggc
      721 accgtggaca accatcactt caagtgcaca tccgagggcg aaggcaagcc ctacgagggc
      781 acccagacca tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc cttcgacatc
      841 ctggctacta gcttcctcta cggcagcaag accttcatca accacaccca gggcatcccc
      901 gacttcttca agcagtcctt ccctgagggc ttcacatggg agagagtcac cacatacgaa
      961 gacgggggcg tgctgaccgc tacccaggac accagcctcc aggacggctg cctcatctac
     1021 aacgtcaaga tcagaggggt gaacttcaca tccaacggcc ctgtgatgca gaagaaaaca
     1081 ctcggctggg aggccttcac cgagacgctg taccccgctg acggcggcct ggaaggcaga
     1141 aacgacatgg ccctgaagct cgtgggcggg agccatctga tcgcaaacat caagaccaca
     1201 tatagatcca agaaacccgc taagaacctc aagatgcctg gcgtctacta tgtggactac
     1261 agactggaaa gaatcaagga ggccaacaac gagacctacg tcgagcagca cgaggtggca
     1321 gtggccagat actgcgacct ccctagcaaa ctggggcaca agcttaattg attttagcgc
     1381 gtgcgccaat tcgaacgctg acgtcatcaa cccgctccaa ggaatcgcgg gcccagtgtc
     1441 actaggcggg aacacccagc gcgcgtgcgc cctggcagga agatggctgt gagggacagg
     1501 ggagtggcgc cctgcaatat ttgcatgtcg ctatgtgttc tgggaaatca ccataaacgt
     1561 gaaatgtctt tggatttggg aatcttataa gttctgtatg agaccacgaa acaccgccat
     1621 ggagaccccg tcccaggttt tagagctaga aatagcaagt taaaataagg ctagtccgtt
     1681 atcaacttga aaaagtggca ccgagtcggt gctttttttg cagacaaatg gctctagagg
     1741 tacccgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     1801 cattgacgtc aatagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     1861 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     1921 ttgacgtcaa tgacggtaaa tggcccgcct ggcattgtgc ccagtacatg accttatggg
     1981 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtcgaggtga
     2041 gccccacgtt ctgcttcact ctccccatct cccccccctc cccaccccca attttgtatt
     2101 tatttatttt ttaattattt tgtgcagcga tgggggcggg gggggggggg gggcgcgcgc
     2161 caggcggggg ggggcggggc gaggggcggg gcggggcgag gcggagaggt gcggcggcag
     2221 ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg cggcggcggc
     2281 cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt cgccccgtgc
     2341 cccgctccgc cgccgcctcg cgccgcccgc cccggctctg actgaccgcg ttactcccac
     2401 aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa ttagctgagc aagaggtaag
     2461 ggtttaaggg atggttggtt ggtggggtat taatgtttaa ttacctggag cacctgcctg
     2521 aaatcacttt ttttcaggtt ggaccggtgc caccatggac tataaggacc acgacggaga
     2581 ctacaaggat catgatattg attacaaaga cgatgacgat aagatggccc caaagaagaa
     2641 gcggaaggtc ggtatccacg gagtcccagc agccgacaag aagtacagca tcggcctggc
     2701 catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa
     2761 gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc
     2821 cctgctgttc gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag
     2881 aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat
     2941 ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga
     3001 taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga
     3061 gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga
     3121 cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat
     3181 cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca
     3241 gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc
     3301 catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc
     3361 cggcgagaag aagaatggcc tgttcggcaa cctgattgcc ctgagcctgg gcctgacccc
     3421 caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac
     3481 ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt
     3541 tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac
     3601 cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca
     3661 ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat
     3721 tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga
     3781 agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct
     3841 cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat
     3901 cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta
     3961 cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta
     4021 ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga
     4081 ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag
     4141 cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa
     4201 gcacagcctg ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt
     4261 gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga
     4321 cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa
     4381 gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc
     4441 cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga
     4501 ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga
     4561 gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca
     4621 gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat
     4681 ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa
     4741 cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa
     4801 agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag
     4861 ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt
     4921 gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac
     4981 ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga
     5041 gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa
     5101 gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat
     5161 caaccggctg tccgactacg atgtggacca tatcgtgcct cagagctttc tgaaggacga
     5221 ctccatcgac aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt
     5281 gccctccgaa gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa
     5341 gctgattacc cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga
     5401 actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca
     5461 cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat
     5521 ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt
     5581 ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa
     5641 cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta
     5701 cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg
     5761 caaggctacc gccaagtact tcttctacag caacatcatg aactttttca agaccgagat
     5821 taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg
     5881 ggagatcgtg tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc
     5941 ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat
     6001 cctgcccaag aggaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa
     6061 gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga
     6121 aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga
     6181 aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt
     6241 gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg
     6301 gaagagaatg ctggcctctg ccggcgaact gcagaaggga aacgaactgg ccctgccctc
     6361 caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga
     6421 ggataatgag cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat
     6481 cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt
     6541 gctgtccgcc tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat
     6601 ccacctgttt accctgacca atctgggagc ccctgccgcc ttcaagtact ttgacaccac
     6661 catcgaccgg aagaggtaca ccagcaccaa agaggtgctg gacgccaccc tgatccacca
     6721 gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag gcgacaaaag
     6781 gccggcggcc acgaaaaagg ccggccaggc aaaaaagaaa aagtaagaat tcctagagct
     6841 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     6901 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     6961 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     7021 agcaaggggg aggattggga agagaatagc aggcatgctg gggagcggcc gcaggaaccc
     7081 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     7141 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     7201 agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     7261 caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg
     7321 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt
     7381 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     7441 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     7501 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     7561 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact
     7621 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa
     7681 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     7741 ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     7801 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     7861 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     7921 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     7981 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     8041 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     8101 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     8161 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     8221 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     8281 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     8341 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     8401 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     8461 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     8521 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     8581 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     8641 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     8701 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     8761 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     8821 ataaatctgg agccggtgag cgtggaagcc gcggtatcat tgcagcactg gggccagatg
     8881 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     8941 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     9001 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     9061 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     9121 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     9181 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     9241 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     9301 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     9361 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     9421 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     9481 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     9541 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     9601 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     9661 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     9721 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     9781 tggccttttg ctggcctttt gctcacatgt
//
