LOCUS       sequence-169987-        8565 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BPNLS-Cas9-BPNLS expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8565)
  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 8565)
  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
FEATURES             Location/Qualifiers
     source          1..8565
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1673
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1597..1619
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1681..1700
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     1728..1747
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1728..1746
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1794..1814
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      1848..1857
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1854..5957
                     /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             1854..5957
                     /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             6003..6023
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6030..6095
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(6120..6139)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6185..6240
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6239..6258)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6644..6661)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6815..7403)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6895..6914)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7574..8365,8366..8434))
                     /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     8197..8216
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8435..8539)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct
     1441 ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc
     1501 ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca gcctcggggc tgtccgcggg
     1561 gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg
     1621 gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg
     1681 gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattctaa tacgactcac
     1741 tatagggaga cgccaccatg aagcggactg ctgatggcag tgaatttgag tccccaaaga
     1801 agaagagaaa ggtggaaggt atccacggag tcccagcagc cgaattcgcc accatggaca
     1861 agaagtattc tatcggactg gacatcggga ctaatagcgt cgggtgggcc gtgatcactg
     1921 acgagtacaa ggtgccctct aagaagttca aggtgctcgg gaacaccgac cggcattcca
     1981 tcaagaaaaa tctgatcgga gctctcctct ttgattcagg ggagaccgct gaagcaaccc
     2041 gcctcaagcg gactgctaga cggcggtaca ccaggaggaa gaaccggatt tgttaccttc
     2101 aagagatatt ctccaacgaa atggcaaagg tcgacgacag cttcttccat aggctggaag
     2161 aatcattcct cgtggaagag gataagaagc atgaacggca tcccatcttc ggtaatatcg
     2221 tcgacgaggt ggcctatcac gagaaatacc caaccatcta ccatcttcgc aaaaagctgg
     2281 tggactcaac cgacaaggca gacctccggc ttatctacct ggccctggcc cacatgatca
     2341 agttcagagg ccacttcctg atcgagggcg acctcaatcc tgacaatagc gatgtggata
     2401 aactgttcat ccagctggtg cagacttaca accagctctt tgaagagaac cccatcaatg
     2461 caagcggagt cgatgccaag gccattctgt cagcccggct gtcaaagagc cgcagacttg
     2521 agaatcttat cgctcagctg ccgggtgaaa agaaaaatgg actgttcggg aacctgattg
     2581 ctctttcact tgggctgact cccaatttca agtctaattt cgacctggca gaggatgcca
     2641 agctgcaact gtccaaggac acctatgatg acgatctcga caacctcctg gcccagatcg
     2701 gtgaccaata cgccgacctt ttccttgctg ctaagaatct ttctgacgcc atcctgctgt
     2761 ctgacattct ccgcgtgaac actgaaatca ccaaggcccc tctttcagct tcaatgatta
     2821 agcggtatga tgagcaccac caggacctga ccctgcttaa ggcactcgtc cggcagcagc
     2881 ttccggagaa gtacaaggaa atcttctttg accagtcaaa gaatggatac gccggctaca
     2941 tcgacggagg tgcctcccaa gaggaatttt ataagtttat caaacctatc cttgagaaga
     3001 tggacggcac cgaagagctc ctcgtgaaac tgaatcggga ggatctgctg cggaagcagc
     3061 gcactttcga caatgggagc attccccacc agatccatct tggggagctt cacgccatcc
     3121 ttcggcgcca agaggacttc tacccctttc ttaaggacaa cagggagaag attgagaaaa
     3181 ttctcacttt ccgcatcccc tactacgtgg gacccctcgc cagaggaaat agccggtttg
     3241 cttggatgac cagaaagtca gaagaaacta tcactccctg gaacttcgaa gaggtggtgg
     3301 acaagggagc cagcgctcag tcattcatcg aacggatgac taacttcgat aagaacctcc
     3361 ccaatgagaa ggtcctgccg aaacattccc tgctctacga gtactttacc gtgtacaacg
     3421 agctgaccaa ggtgaaatat gtcaccgaag ggatgaggaa gcccgcattc ctgtcaggcg
     3481 aacaaaagaa ggcaattgtg gaccttctgt tcaagaccaa tagaaaggtg accgtgaagc
     3541 agctgaagga ggactatttc aagaaaattg aatgcttcga ctctgtggag attagcgggg
     3601 tcgaagatcg gttcaacgca agcctgggta cctaccatga tctgcttaag atcatcaagg
     3661 acaaggattt tctggacaat gaggagaacg aggacatcct tgaggacatt gtcctgactc
     3721 tcactctgtt cgaggaccgg gaaatgatcg aggagaggct taagacctac gcccatctgt
     3781 tcgacgataa agtgatgaag caacttaaac ggagaagata taccggatgg ggacgcctta
     3841 gccgcaaact catcaacgga atccgggaca aacagagcgg aaagaccatt cttgatttcc
     3901 ttaagagcga cggattcgct aatcgcaact tcatgcaact tatccatgat gattccctga
     3961 cctttaagga ggacatccag aaggcccaag tgtctggaca aggtgactca ctgcacgagc
     4021 atatcgcaaa tctggctggt tcacccgcta ttaagaaggg tattctccag accgtgaaag
     4081 tcgtggacga gctggtcaag gtgatgggtc gccataaacc agagaacatt gtcatcgaga
     4141 tggccaggga aaaccagact acccagaagg gacagaagaa cagcagggag cggatgaaaa
     4201 gaattgagga agggattaag gagctcgggt cacagatcct taaagagcac ccggtggaaa
     4261 acacccagct tcagaatgag aagctctatc tgtactacct tcaaaatgga cgcgatatgt
     4321 atgtggacca agagcttgat atcaacaggc tctcagacta cgacgtggac cacatcgtcc
     4381 ctcagagctt cctcaaagac gactcaattg acaataaggt gctgactcgc tcagacaaga
     4441 accggggaaa gtcagataac gtgccctcag aggaagtcgt gaaaaagatg aagaactatt
     4501 ggcgccagct tctgaacgca aagctgatca ctcagcggaa gttcgacaat ctcactaagg
     4561 ctgagagggg cggactgagc gaactggaca aagcaggatt cattaaacgg caacttgtgg
     4621 agactcggca gattactaaa catgtcgccc aaatccttga ctcacgcatg aataccaagt
     4681 acgacgaaaa cgacaaactt atccgcgagg tgaaggtgat taccctgaag tccaagctgg
     4741 tcagcgattt cagaaaggac tttcaattct acaaagtgcg ggagatcaat aactatcatc
     4801 atgctcatga cgcatatctg aatgccgtgg tgggaaccgc cctgatcaag aagtacccaa
     4861 agctggaaag cgagttcgtg tacggagact acaaggtcta cgacgtgcgc aagatgattg
     4921 ccaaatctga gcaggagatc ggaaaggcca ccgcaaagta cttcttctac agcaacatca
     4981 tgaatttctt caagaccgaa atcacccttg caaacggtga gatccggaag aggccgctca
     5041 tcgagactaa tggggagact ggcgaaatcg tgtgggacaa gggcagagat ttcgctaccg
     5101 tgcgcaaagt gctttctatg cctcaagtga acatcgtgaa gaaaaccgag gtgcaaaccg
     5161 gaggcttttc taaggaatca atcctcccca agcgcaactc cgacaagctc attgcaagga
     5221 agaaggattg ggaccctaag aagtacggcg gattcgattc accaactgtg gcttattctg
     5281 tcctggtcgt ggctaaggtg gaaaaaggaa agtctaagaa gctcaagagc gtgaaggaac
     5341 tgctgggtat caccattatg gagcgcagct ccttcgagaa gaacccaatt gactttctcg
     5401 aagccaaagg ttacaaggaa gtcaagaagg accttatcat caagctccca aagtatagcc
     5461 tgttcgaact ggagaatggg cggaagcgga tgctcgcctc cgctggcgaa cttcagaagg
     5521 gtaatgagct ggctctcccc tccaagtacg tgaatttcct ctaccttgca agccattacg
     5581 agaagctgaa ggggagcccc gaggacaacg agcaaaagca actgtttgtg gagcagcata
     5641 agcattatct ggacgagatc attgagcaga tttccgagtt ttctaaacgc gtcattctcg
     5701 ctgatgccaa cctcgataaa gtccttagcg catacaataa gcacagagac aaaccaattc
     5761 gggagcaggc tgagaatatc atccacctgt tcaccctcac caatcttggt gcccctgccg
     5821 cattcaagta cttcgacacc accatcgacc ggaaacgcta tacctccacc aaagaagtgc
     5881 tggacgccac cctcatccac cagagcatca ccggacttta cgaaactcgg attgacctct
     5941 cacagctcgg aggggatgag ggagctgata agcgcaccgc cgatggttcc gagttcgaaa
     6001 gccccaaaaa aaagcgcaag gtcgaagatg actataagga tcacgacggt gattataaag
     6061 accacgatat cgactataaa gacgacgatg acaaagagct gtataaactg ctgtaaagat
     6121 ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct
     6181 ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca
     6241 ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag
     6301 agtttggcaa catatgccat atgctggctg ccatgaacaa aggtggctat aaagaggtca
     6361 tcagtatatg aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg
     6421 aggttagatt ttttttatat tttgttttgt gttatttttt tctttaacat ccctaaaatt
     6481 ttccttacat gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc
     6541 tgtccctctt ctcttatgaa gatccctcga ccgatccgct gcattaatga atcggccaac
     6601 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     6661 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     6721 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     6781 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     6841 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     6901 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     6961 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct
     7021 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     7081 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     7141 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     7201 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     7261 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     7321 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     7381 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     7441 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     7501 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     7561 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     7621 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     7681 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     7741 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     7801 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     7861 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     7921 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     7981 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     8041 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     8101 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     8161 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     8221 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     8281 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     8341 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     8401 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     8461 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     8521 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctg
//
