LOCUS       sequence-469881-        8971 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Cas9 and two guideRNAs, one to cut the genome downstream of 
            HSPA5 (BiP/GRP78) and one to linearize the DONOR plasmid..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8971)
  TITLE     Rivera-Fuentes plasmids March 2025
REFERENCE   2  (bases 1 to 8971)
  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"
COMMENT     Sequence Label: CRISPR plasmid
FEATURES             Location/Qualifiers
     source          1..8971
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(2946..2966)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(2973..3038)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      3038..3047
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     intron          3059..3286
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     enhancer        3587..3872
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     misc_RNA        complement(3969..4044)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(4065..4313)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        complement(4411..4486)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(4508..4756)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     rep_origin      complement(4818..5406)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4898..4917)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5577..6368,6369..6437))
                     /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     6200..6219
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6438..6542)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6610..6628
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6666..6688)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6788..6807
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6824..7279)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6961..6982)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7173..7192
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   7354..7494
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   7365..7494
                     /label=AAV2 ITR
     polyA_signal    7503..7710
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     7699..7716
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(7744..7791)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(join(7792..8971,1..2921))
                     /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             complement(join(7792..8971,1..2921))
                     /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"
ORIGIN
        1 actttgtaaa actggaaatc cttccggaaa tcggacacca gcttggactt cagggtgatc
       61 actttcactt cccggatcag cttgtcattc tcgtcgtact tagtgttcat ccgggagtcc
      121 aggatctgtg ccacgtgctt tgtgatctgc cgggtttcca ccagctgtct cttgatgaag
      181 ccggccttat ccagttcgct caggccgcct ctctcggcct tggtcagatt gtcgaacttt
      241 ctctgggtaa tcagcttggc gttcagcagc tgccgccagt agttcttcat cttcttcacg
      301 acctcttcgg agggcacgtt gtcgctcttg ccccggttct tgtcgcttct ggtcagcacc
      361 ttgttgtcga tggagtcgtc cttcagaaag ctctgaggca cgatatggtc cacatcgtag
      421 tcggacagcc ggttgatgtc cagttcctgg tccacgtaca tatcccgccc attctgcagg
      481 tagtacaggt acagcttctc gttctgcagc tgggtgtttt ccacggggtg ttctttcagg
      541 atctggctgc ccagctcttt gatgccctct tcgatccgct tcattctctc gcggctgttc
      601 ttctgtccct tctgggtggt ctggttctct ctggccattt cgatcacgat gttctcgggc
      661 ttgtgccggc ccatcacttt cacgagctcg tccaccacct tcactgtctg caggatgccc
      721 ttcttaatgg cggggctgcc ggccagattg gcaatgtgct cgtgcaggct atcgccctgg
      781 ccggacacct gggctttctg gatgtcctct ttaaaggtca ggctgtcgtc gtggatcagc
      841 tgcatgaagt ttctgttggc gaagccgtcg gacttcagga aatccaggat tgtcttgccg
      901 gactgcttgt cccggatgcc gttgatcagc ttccggctca gcctgcccca gccggtgtat
      961 ctccgccgct tcagctgctt catcactttg tcgtcgaaca ggtgggcata ggttttcagc
     1021 cgttcctcga tcatctctct gtcctcaaac agtgtcaggg tcagcacgat atcttccaga
     1081 atgtcctcgt tttcctcatt gtccaggaag tccttgtcct tgataatttt cagcagatcg
     1141 tggtatgtgc ccagggaggc gttgaaccga tcttccacgc cggagatttc cacggagtcg
     1201 aagcactcga ttttcttgaa gtagtcctct ttcagctgct tcacggtcac tttccggttg
     1261 gtcttgaaca gcaggtccac gatggccttt ttctgctcgc cgctcaggaa ggcgggcttt
     1321 ctcattccct cggtcacgta tttcactttg gtcagctcgt tatacacggt gaagtactcg
     1381 tacagcaggc tgtgcttggg cagcaccttc tcgttgggca ggttcttatc gaagttggtc
     1441 atccgctcga tgaagctctg ggcggaagcg cccttgtcca ccacttcctc gaagttccag
     1501 ggggtgatgg tttcctcgct ctttctggtc atccaggcga atctgctgtt tcccctggcc
     1561 agagggccca cgtagtaggg gatgcggaag gtcaggatct tctcgatctt ttcccggttg
     1621 tccttcagga atgggtaaaa atcttcctgc cgccgcagaa tggcgtgcag ctctcccagg
     1681 tggatctggt gggggatgct gccgttgtcg aaggtccgct gcttccgcag caggtcctct
     1741 ctgttcagct tcacgagcag ttcctcggtg ccgtccatct tttccaggat gggcttgatg
     1801 aacttgtaga actcttcctg gctggctccg ccgtcaatgt agccggcgta gccgttcttg
     1861 ctctggtcga agaaaatctc tttgtacttc tcaggcagct gctgccgcac gagagctttc
     1921 agcagggtca ggtcctggtg gtgctcgtcg tatctcttga tcatagaggc gctcaggggg
     1981 gccttggtga tctcggtgtt cactctcagg atgtcgctca gcaggatggc gtcggacagg
     2041 ttcttggcgg ccagaaacag gtcggcgtac tggtcgccga tctgggccag caggttgtcc
     2101 aggtcgtcgt cgtaggtgtc cttgctcagc tgcagtttgg catcctcggc caggtcgaag
     2161 ttgctcttga agttgggggt caggcccagg ctcagggcaa tcaggtttcc gaacaggcca
     2221 ttcttcttct cgccgggcag ctgggcgatc agattttcca gccgtctgct cttgctcagt
     2281 ctggcagaca ggatggcctt ggcgtccacg ccgctggcgt tgatggggtt ttcctcgaac
     2341 agctggttgt aggtctgcac cagctggatg aacagcttgt ccacgtcgct gttgtcgggg
     2401 ttcaggtcgc cctcgatcag gaagtggccc cggaacttga tcatgtgggc cagggccaga
     2461 tagatcagcc gcaggtcggc cttgtcggtg ctgtccacca gtttctttct caggtggtag
     2521 atggtggggt acttctcgtg gtaggccacc tcgtccacga tgttgccgaa gatggggtgc
     2581 cgctcgtgct tcttatcctc ttccaccagg aaggactctt ccagtctgtg gaagaagctg
     2641 tcgtccacct tggccatctc gttgctgaag atctcttgca gatagcagat ccggttcttc
     2701 cgtctggtgt atcttcttct ggcggttctc ttcagccggg tggcctcggc tgtttcgccg
     2761 ctgtcgaaca gcagggctcc gatcaggttc ttcttgatgc tgtgccggtc ggtgttgccc
     2821 agcaccttga atttcttgct gggcaccttg tactcgtcgg tgatcacggc ccagcccaca
     2881 gagttggtgc cgatgtccag gccgatgctg tacttcttgt cggctgctgg gactccgtgg
     2941 ataccgacct tccgcttctt ctttggggcc atcttatcgt catcgtcttt gtaatcaata
     3001 tcatgatcct tgtagtctcc gtcgtggtcc ttatagtcca tggtggcacc ggtccaacct
     3061 gaaaaaaagt gatttcaggc aggtgctcca ggtaattaaa cattaatacc ccaccaacca
     3121 accatccctt aaacccttac ctcttgctca gctaattaca gcccggagga gaagggccgt
     3181 cccgcccgct cacctgtggg agtaacgcgg tcagtcagag ccggggcggg cggcgcgagg
     3241 cggcggcgga gcggggcacg gggcgaaggc agcgcgcagc gactcccgcc cgccgcgcgc
     3301 ttcgcttttt atagggccgc cgccgccgcc gcctcgccat aaaaggaaac tttcggagcg
     3361 cgccgctctg attggctgcc gccgcacctc tccgcctcgc cccgcccccc cccccccccc
     3421 cccccccccc cccccccccc cccccccccc cccgcccggc gcggcccccc cccccccccg
     3481 cccccatcgc tgcacaaaat aattaaaaaa taaataaata caaaattggg ggtggggagg
     3541 ggggggagat ggggagagtg aagcagaacg tggggctcac ctcgaccatg gtaatagcga
     3601 tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg tactgggcac
     3661 aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt ggcatatgat
     3721 acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat
     3781 ggaaagtccc tattggcgtt actattgacg tcaatgggcg ggggtcgttg ggcggtcagc
     3841 caggcgggcc atttaccgta agttatgtaa cgggtacctc tagagccatt tgtctgcaga
     3901 attggcgcac gcgctaaaaa cggactagcc ttattttaac ttgctatttc tagctctaaa
     3961 acaaaaaagc accgactcgg tgccactttt tcaagttgat aacggactag ccttatttta
     4021 acttgctatt tctagctcta aaacaacacg tacgcgtacg atgcggtgtt tcgtcctttc
     4081 cacaagatat ataaagccaa gaaatcgaaa tactttcaag ttacggtaag catatgatag
     4141 tccattttaa aacataattt taaaactgca aactacccaa gaaattatta ctttctacgt
     4201 cacgtatttt gtactaatat ctttgtgttt acagtcaaat taattccaat tatctctcta
     4261 acagccttgt atcgtatatg caaatatgaa ggaatcatgg gaaataggcc ctcacatgcc
     4321 tctagagcca tttgtctgca gaattggcgc acgcgctaaa aacggactag ccttatttta
     4381 acttgctatt tctagctcta aaacaaaaaa gcaccgactc ggtgccactt tttcaagttg
     4441 ataacggact agccttattt taacttgcta tttctagctc taaaacctta ggctatagca
     4501 gtttcacggt gtttcgtcct ttccacaaga tatataaagc caagaaatcg aaatactttc
     4561 aagttacggt aagcatatga tagtccattt taaaacataa ttttaaaact gcaaactacc
     4621 caagaaatta ttactttcta cgtcacgtat tttgtactaa tatctttgtg tttacagtca
     4681 aattaattcc aattatctct ctaacagcct tgtatcgtat atgcaaatat gaaggaatca
     4741 tgggaaatag gccctcacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4801 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4861 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4921 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4981 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5041 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5101 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5161 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5221 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     5281 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5341 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5401 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5461 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5521 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5581 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5641 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     5701 gctgcaatga taccgcggct tccacgctca ccggctccag atttatcagc aataaaccag
     5761 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     5821 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     5881 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     5941 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6001 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6061 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6121 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6181 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6241 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6301 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6361 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6421 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6481 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6541 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     6601 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     6661 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     6721 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     6781 aactatgcgg catcagagca gattgtactg agagtgcacc ataaaattgt aaacgttaat
     6841 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaatagacc
     6901 gaaatcggca aaatccctta taaatcaaaa gaatagcccg agatagagtt gagtgttgtt
     6961 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     7021 accgtctatc agggcgatgg cccactacgt gaaccatcac ccaaatcaag ttttttgggg
     7081 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     7141 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     7201 aaggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     7261 gcgccgctac agggcgcgta ctatggttgc tttgacgtat gcggtgtgaa ataccgcaca
     7321 gatgcgtaag gagaaaatac cgcatcaggc gcccctgcag gcagctgcgc gctcgctcgc
     7381 tcactgaggc cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag
     7441 tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctgcggcc
     7501 gctccccagc atgcctgcta ttctcttccc aatcctcccc cttgctgtcc tgccccaccc
     7561 caccccccag aatagaatga cacctactca gacaatgcga tgcaatttcc tcattttatt
     7621 aggaaaggac agtgggagtg gcaccttcca gggtcaagga aggcacgggg gaggggcaaa
     7681 caacagatgg ctggcaacta gaaggcacag tcgaggctga tcagcgagct ctaggaattc
     7741 ttactttttc ttttttgcct ggccggcctt tttcgtggcc gccggccttt tgtcgcctcc
     7801 cagctgagac aggtcgatcc gtgtctcgta caggccggtg atgctctggt ggatcagggt
     7861 ggcgtccagc acctctttgg tgctggtgta cctcttccgg tcgatggtgg tgtcaaagta
     7921 cttgaaggcg gcaggggctc ccagattggt cagggtaaac aggtggatga tattctcggc
     7981 ctgctctctg atgggcttat cccggtgctt gttgtaggcg gacagcactt tgtccagatt
     8041 agcgtcggcc aggatcactc tcttggagaa ctcgctgatc tgctcgatga tctcgtccag
     8101 gtagtgcttg tgctgttcca caaacagctg tttctgctca ttatcctcgg gggagccctt
     8161 cagcttctca tagtggctgg ccaggtacag gaagttcaca tatttggagg gcagggccag
     8221 ttcgtttccc ttctgcagtt cgccggcaga ggccagcatt ctcttccggc cgttttccag
     8281 ctcgaacagg gagtacttag gcagcttgat gatcaggtcc tttttcactt ctttgtagcc
     8341 cttggcttcc agaaagtcga tgggattctt ctcgaagctg cttctttcca tgatggtgat
     8401 ccccagcagc tctttcacac tcttcagttt cttggacttg cccttttcca ctttggccac
     8461 caccagcaca gaataggcca cggtggggct gtcgaagccg ccgtacttct tagggtccca
     8521 gtccttcttt ctggcgatca gcttatcgct gttcctcttg ggcaggatag actctttgct
     8581 gaagccgcct gtctgcacct cggtcttttt cacgatattc acttggggca tgctcagcac
     8641 tttccgcacg gtggcaaaat cccggccctt atcccacacg atctccccgg tttcgccgtt
     8701 tgtctcgatc agaggccgct tccggatctc gccgttggcc agggtaatct cggtcttgaa
     8761 aaagttcatg atgttgctgt agaagaagta cttggcggta gccttgccga tttcctgctc
     8821 gctcttggcg atcatcttcc gcacgtcgta caccttgtag tcgccgtaca cgaactcgct
     8881 ttccagctta gggtactttt tgatcagggc ggttcccacg acggcgttca ggtaggcgtc
     8941 gtgggcgtgg tggtagttgt tgatctcgcg c
//
