LOCUS       sequence-174888-        8567 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  BPNLS-Cas9-BPNLS-2AGFP expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8567)
  AUTHORS   Suzuki K, Tsunekawa Y, Hernandez-Benitez R, Wu J, Zhu J, Kim EJ, 
            Hatanaka F, Yamamoto M, Araoka T, Li Z, Kurita M, Hishida T, Li M, 
            Aizawa E, Guo S, Chen S, Goebl A, Soligalla RD, Qu J, Jiang T, Fu X,
            Jafari M, Esteban CR, Berggren WT, Lajara J, Nunez-Delicado E, 
            Guillen P, Campistol JM, Matsuzaki F, Liu GH, Magistretti P, Zhang 
            K, Callaway EM, Zhang K, Belmonte JC
  TITLE     In vivo genome editing via CRISPR/Cas9 mediated homology-independent
            targeted integration.
  JOURNAL   Nature. 2016 Dec 1;540(7631):144-149. doi: 10.1038/nature20565. Epub
            2016 Nov 16.
  PUBMED    27851729
REFERENCE   2  (bases 1 to 8567)
  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: 
            "10.1038/nature20565"; journalName: "Nature"; date: "2016-12-1- 1"; 
            volume: "540"; issue: "7631"; pages: "144-149"
COMMENT     Sequence Label: pCAG-1BPNLS-Cas9-1BPNLS-2AGFP
FEATURES             Location/Qualifiers
     source          1..8567
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     326..348
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     410..429
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     457..476
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        457..475
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             523..543
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      577..586
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             583..4686
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             583..4686
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             4732..4752
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4759..4824
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4846..4899
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(4900..4902,4903..5616)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(4900..4902,4903..5613)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(4942..4963)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5203..5222)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5550..5571
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5620..5639)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5685..5740
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5739..5758)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6146..6163)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6317..6905)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6397..6416)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7076..7867,7868..7936))
                     /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     7699..7718
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7937..8041)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        8072..8451
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggggagggct cgggggaggg gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg
       61 gcgagccgca gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg
      121 tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc
      181 ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg
      241 ccgcgccgcc gtccccttct ccctctccag cctcggggct gtccgcgggg ggacggctgc
      301 cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg cggctctaga
      361 gcctctgcta accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg
      421 gttattgtgc tgtctcatca ttttggcaaa gaattctaat acgactcact atagggagac
      481 gccaccatga agcggactgc tgatggcagt gaatttgagt ccccaaagaa gaagagaaag
      541 gtggaaggta tccacggagt cccagcagcc gaattcgcca ccatggacaa gaagtattct
      601 atcggactgg acatcgggac taatagcgtc gggtgggccg tgatcactga cgagtacaag
      661 gtgccctcta agaagttcaa ggtgctcggg aacaccgacc ggcattccat caagaaaaat
      721 ctgatcggag ctctcctctt tgattcaggg gagaccgctg aagcaacccg cctcaagcgg
      781 actgctagac ggcggtacac caggaggaag aaccggattt gttaccttca agagatattc
      841 tccaacgaaa tggcaaaggt cgacgacagc ttcttccata ggctggaaga atcattcctc
      901 gtggaagagg ataagaagca tgaacggcat cccatcttcg gtaatatcgt cgacgaggtg
      961 gcctatcacg agaaataccc aaccatctac catcttcgca aaaagctggt ggactcaacc
     1021 gacaaggcag acctccggct tatctacctg gccctggccc acatgatcaa gttcagaggc
     1081 cacttcctga tcgagggcga cctcaatcct gacaatagcg atgtggataa actgttcatc
     1141 cagctggtgc agacttacaa ccagctcttt gaagagaacc ccatcaatgc aagcggagtc
     1201 gatgccaagg ccattctgtc agcccggctg tcaaagagcc gcagacttga gaatcttatc
     1261 gctcagctgc cgggtgaaaa gaaaaatgga ctgttcggga acctgattgc tctttcactt
     1321 gggctgactc ccaatttcaa gtctaatttc gacctggcag aggatgccaa gctgcaactg
     1381 tccaaggaca cctatgatga cgatctcgac aacctcctgg cccagatcgg tgaccaatac
     1441 gccgaccttt tccttgctgc taagaatctt tctgacgcca tcctgctgtc tgacattctc
     1501 cgcgtgaaca ctgaaatcac caaggcccct ctttcagctt caatgattaa gcggtatgat
     1561 gagcaccacc aggacctgac cctgcttaag gcactcgtcc ggcagcagct tccggagaag
     1621 tacaaggaaa tcttctttga ccagtcaaag aatggatacg ccggctacat cgacggaggt
     1681 gcctcccaag aggaatttta taagtttatc aaacctatcc ttgagaagat ggacggcacc
     1741 gaagagctcc tcgtgaaact gaatcgggag gatctgctgc ggaagcagcg cactttcgac
     1801 aatgggagca ttccccacca gatccatctt ggggagcttc acgccatcct tcggcgccaa
     1861 gaggacttct acccctttct taaggacaac agggagaaga ttgagaaaat tctcactttc
     1921 cgcatcccct actacgtggg acccctcgcc agaggaaata gccggtttgc ttggatgacc
     1981 agaaagtcag aagaaactat cactccctgg aacttcgaag aggtggtgga caagggagcc
     2041 agcgctcagt cattcatcga acggatgact aacttcgata agaacctccc caatgagaag
     2101 gtcctgccga aacattccct gctctacgag tactttaccg tgtacaacga gctgaccaag
     2161 gtgaaatatg tcaccgaagg gatgaggaag cccgcattcc tgtcaggcga acaaaagaag
     2221 gcaattgtgg accttctgtt caagaccaat agaaaggtga ccgtgaagca gctgaaggag
     2281 gactatttca agaaaattga atgcttcgac tctgtggaga ttagcggggt cgaagatcgg
     2341 ttcaacgcaa gcctgggtac ctaccatgat ctgcttaaga tcatcaagga caaggatttt
     2401 ctggacaatg aggagaacga ggacatcctt gaggacattg tcctgactct cactctgttc
     2461 gaggaccggg aaatgatcga ggagaggctt aagacctacg cccatctgtt cgacgataaa
     2521 gtgatgaagc aacttaaacg gagaagatat accggatggg gacgccttag ccgcaaactc
     2581 atcaacggaa tccgggacaa acagagcgga aagaccattc ttgatttcct taagagcgac
     2641 ggattcgcta atcgcaactt catgcaactt atccatgatg attccctgac ctttaaggag
     2701 gacatccaga aggcccaagt gtctggacaa ggtgactcac tgcacgagca tatcgcaaat
     2761 ctggctggtt cacccgctat taagaagggt attctccaga ccgtgaaagt cgtggacgag
     2821 ctggtcaagg tgatgggtcg ccataaacca gagaacattg tcatcgagat ggccagggaa
     2881 aaccagacta cccagaaggg acagaagaac agcagggagc ggatgaaaag aattgaggaa
     2941 gggattaagg agctcgggtc acagatcctt aaagagcacc cggtggaaaa cacccagctt
     3001 cagaatgaga agctctatct gtactacctt caaaatggac gcgatatgta tgtggaccaa
     3061 gagcttgata tcaacaggct ctcagactac gacgtggacc acatcgtccc tcagagcttc
     3121 ctcaaagacg actcaattga caataaggtg ctgactcgct cagacaagaa ccggggaaag
     3181 tcagataacg tgccctcaga ggaagtcgtg aaaaagatga agaactattg gcgccagctt
     3241 ctgaacgcaa agctgatcac tcagcggaag ttcgacaatc tcactaaggc tgagaggggc
     3301 ggactgagcg aactggacaa agcaggattc attaaacggc aacttgtgga gactcggcag
     3361 attactaaac atgtcgccca aatccttgac tcacgcatga ataccaagta cgacgaaaac
     3421 gacaaactta tccgcgaggt gaaggtgatt accctgaagt ccaagctggt cagcgatttc
     3481 agaaaggact ttcaattcta caaagtgcgg gagatcaata actatcatca tgctcatgac
     3541 gcatatctga atgccgtggt gggaaccgcc ctgatcaaga agtacccaaa gctggaaagc
     3601 gagttcgtgt acggagacta caaggtctac gacgtgcgca agatgattgc caaatctgag
     3661 caggagatcg gaaaggccac cgcaaagtac ttcttctaca gcaacatcat gaatttcttc
     3721 aagaccgaaa tcacccttgc aaacggtgag atccggaaga ggccgctcat cgagactaat
     3781 ggggagactg gcgaaatcgt gtgggacaag ggcagagatt tcgctaccgt gcgcaaagtg
     3841 ctttctatgc ctcaagtgaa catcgtgaag aaaaccgagg tgcaaaccgg aggcttttct
     3901 aaggaatcaa tcctccccaa gcgcaactcc gacaagctca ttgcaaggaa gaaggattgg
     3961 gaccctaaga agtacggcgg attcgattca ccaactgtgg cttattctgt cctggtcgtg
     4021 gctaaggtgg aaaaaggaaa gtctaagaag ctcaagagcg tgaaggaact gctgggtatc
     4081 accattatgg agcgcagctc cttcgagaag aacccaattg actttctcga agccaaaggt
     4141 tacaaggaag tcaagaagga ccttatcatc aagctcccaa agtatagcct gttcgaactg
     4201 gagaatgggc ggaagcggat gctcgcctcc gctggcgaac ttcagaaggg taatgagctg
     4261 gctctcccct ccaagtacgt gaatttcctc taccttgcaa gccattacga gaagctgaag
     4321 gggagccccg aggacaacga gcaaaagcaa ctgtttgtgg agcagcataa gcattatctg
     4381 gacgagatca ttgagcagat ttccgagttt tctaaacgcg tcattctcgc tgatgccaac
     4441 ctcgataaag tccttagcgc atacaataag cacagagaca aaccaattcg ggagcaggct
     4501 gagaatatca tccacctgtt caccctcacc aatcttggtg cccctgccgc attcaagtac
     4561 ttcgacacca ccatcgaccg gaaacgctat acctccacca aagaagtgct ggacgccacc
     4621 ctcatccacc agagcatcac cggactttac gaaactcgga ttgacctctc acagctcgga
     4681 ggggatgagg gagctgataa gcgcaccgcc gatggttccg agttcgaaag ccccaaaaaa
     4741 aagcgcaagg tcgaagatga ctataaggat cacgacggtg attataaaga ccacgatatc
     4801 gactataaag acgacgatga caaagagctg tataaactgc tgttcgaagg aaggggctct
     4861 ttgcttactt gtggagatgt tgaggaaaat ccaggacccg tgagcaaggg cgaggagctg
     4921 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     4981 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     5041 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     5101 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     5161 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     5221 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     5281 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     5341 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     5401 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     5461 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     5521 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     5581 gggatcactc tcggcatgga cgagctgtac aagtgaagat ctttttccct ctgccaaaaa
     5641 ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat
     5701 tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag
     5761 ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccca
     5821 tatgctggct gccatgaaca aaggttggct ataaagaggt catcagtata tgaaacagcc
     5881 ccctgctgtc cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat
     5941 attttgtttt gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact
     6001 agccagattt ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg
     6061 gagatccctc gaccgatccg ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     6121 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     6181 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     6241 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6301 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6361 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6421 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6481 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     6541 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6601 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6661 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6721 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     6781 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6841 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     6901 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     6961 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     7021 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     7081 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     7141 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     7201 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     7261 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     7321 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     7381 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     7441 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     7501 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     7561 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     7621 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     7681 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     7741 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     7801 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7861 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     7921 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     7981 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     8041 gcacatttcc ccgaaaagtg ccacctgggt cgacattgat tattgactag ttattaatag
     8101 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     8161 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     8221 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     8281 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     8341 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     8401 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtcgaggtg
     8461 agccccacgt tctgcttcac tctccccatc tcccccccct ccccaccccc aattttgtat
     8521 ttatttattt tttaattatt ttgtgcagcg atgggggcgg ggggggg
//
