LOCUS       sequence-267592-        8569 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector for sgRNAs cloned into the BsaI sites and for 
            expression of Cas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8569)
  AUTHORS   Reuven N, Adler J, Broennimann K, Myers N, Shaul Y
  TITLE     Recruitment of DNA Repair MRN Complex by Intrinsically Disordered 
            Protein Domain Fused to Cas9 Improves Efficiency of CRISPR-Mediated 
            Genome Editing.
  JOURNAL   Biomolecules. 2019 Oct 8;9(10). pii: biom9100584. doi: 
            10.3390/biom9100584.
  PUBMED    31597252
REFERENCE   2  (bases 1 to 8569)
  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"; journalName: 
            "Biomolecules."; date: "2019-10-8"; volume: "9(10). pii: 
            biom9100584. doi"; pages: " 10.3390/biom9100584"
COMMENT     Sequence Label: pU6-(BsaI)_CBh-Cas9
FEATURES             Location/Qualifiers
     source          1..8569
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          143..370
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      382..391
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             391..456
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             463..483
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             508..4608
                     /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             508..4608
                     /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             4609..4656
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(4684..4701)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4690..4897
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4906..5046
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4906..5035
                     /label=AAV2 ITR
     rep_origin      5121..5576
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5208..5227)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5418..5439
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5593..5612)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5712..5734
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5772..5790)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5858..5962
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5963..6031,6032..6823)
                     /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(6181..6200)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6994..7582
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7483..7502
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        7644..7892
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7644..7664
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7815..7834
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        7913..7988
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8085..8370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 ggcggggcgg ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg cagccaatca
       61 gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa
      121 aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc
      181 cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag
      241 cgggcgggac ggcccttctc ctccgggctg taattagctg agcaagaggt aagggtttaa
      301 gggatggttg gttggtgggg tattaatgtt taattacctg gagcacctgc ctgaaatcac
      361 tttttttcag gttggaccgg tgccaccatg gactataagg accacgacgg agactacaag
      421 gatcatgata ttgattacaa agacgatgac gataagatgg ccccaaagaa gaagcggaag
      481 gtcggtatcc acggagtccc agcagccgac aagaagtaca gcatcggcct ggacatcggc
      541 accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag caagaaattc
      601 aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg agccctgctg
      661 ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga gaaccgccag aagaagatac
      721 accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga gatggccaag
      781 gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga ggataagaag
      841 cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac
      901 cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc cgacctgcgg
      961 ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct gatcgagggc
     1021 gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt gcagacctac
     1081 aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa ggccatcctg
     1141 tctgccagac tgagcaagag cagacggctg gaaaatctga tcgcccagct gcccggcgag
     1201 aagaagaatg gcctgttcgg aaacctgatt gccctgagcc tgggcctgac ccccaacttc
     1261 aagagcaact tcgacctggc cgaggatgcc aaactgcagc tgagcaagga cacctacgac
     1321 gacgacctgg acaacctgct ggcccagatc ggcgaccagt acgccgacct gtttctggcc
     1381 gccaagaacc tgtccgacgc catcctgctg agcgacatcc tgagagtgaa caccgagatc
     1441 accaaggccc ccctgagcgc ctctatgatc aagagatacg acgagcacca ccaggacctg
     1501 accctgctga aagctctcgt gcggcagcag ctgcctgaga agtacaaaga gattttcttc
     1561 gaccagagca agaacggcta cgccggctac attgacggcg gagccagcca ggaagagttc
     1621 tacaagttca tcaagcccat cctggaaaag atggacggca ccgaggaact gctcgtgaag
     1681 ctgaacagag aggacctgct gcggaagcag cggaccttcg acaacggcag catcccccac
     1741 cagatccacc tgggagagct gcacgccatt ctgcggcggc aggaagattt ttacccattc
     1801 ctgaaggaca accgggaaaa gatcgagaag atcctgacct tccgcatccc ctactacgtg
     1861 ggccctctgg ccaggggaaa cagcagattc gcctggatga ccagaaagag cgaggaaacc
     1921 atcaccccct ggaacttcga ggaagtggtg gacaagggcg cttccgccca gagcttcatc
     1981 gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc caagcacagc
     2041 ctgctgtacg agtacttcac cgtgtataac gagctgacca aagtgaaata cgtgaccgag
     2101 ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa aggccatcgt ggacctgctg
     2161 ttcaagacca accggaaagt gaccgtgaag cagctgaaag aggactactt caagaaaatc
     2221 gagtgcttcg actccgtgga aatctccggc gtggaagatc ggttcaacgc ctccctgggc
     2281 acataccacg atctgctgaa aattatcaag gacaaggact tcctggacaa tgaggaaaac
     2341 gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag agagatgatc
     2401 gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag
     2461 cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg catccgggac
     2521 aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc caacagaaac
     2581 ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca gaaagcccag
     2641 gtgtccggcc agggcgatag cctgcacgag cacattgcca atctggccgg cagccccgcc
     2701 attaagaagg gcatcctgca gacagtgaag gtggtggacg agctcgtgaa agtgatgggc
     2761 cggcacaagc ccgagaacat cgtgatcgaa atggccagag agaaccagac cacccagaag
     2821 ggacagaaga acagccgcga gagaatgaag cggatcgaag agggcatcaa agagctgggc
     2881 agccagatcc tgaaagaaca ccccgtggaa aacacccagc tgcagaacga gaagctgtac
     2941 ctgtactacc tgcagaatgg gcgggatatg tacgtggacc aggaactgga catcaaccgg
     3001 ctgtccgact acgatgtgga ccatatcgtg cctcagagct ttctgaagga cgactccatc
     3061 gacaacaagg tgctgaccag aagcgacaag aaccggggca agagcgacaa cgtgccctcc
     3121 gaagaggtcg tgaagaagat gaagaactac tggcggcagc tgctgaacgc caagctgatt
     3181 acccagagaa agttcgacaa tctgaccaag gccgagagag gcggcctgag cgaactggat
     3241 aaggccggct tcatcaagag acagctggtg gaaacccggc agatcacaaa gcacgtggca
     3301 cagatcctgg actcccggat gaacactaag tacgacgaga atgacaagct gatccgggaa
     3361 gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccggaagga tttccagttt
     3421 tacaaagtgc gcgagatcaa caactaccac cacgcccacg acgcctacct gaacgccgtc
     3481 gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt gtacggcgac
     3541 tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg agcaggaaat cggcaaggct
     3601 accgccaagt acttcttcta cagcaacatc atgaactttt tcaagaccga gattaccctg
     3661 gccaacggcg agatccggaa gcggcctctg atcgagacaa acggcgaaac cggggagatc
     3721 gtgtgggata agggccggga ttttgccacc gtgcggaaag tgctgagcat gccccaagtg
     3781 aatatcgtga aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc
     3841 aagaggaaca gcgataagct gatcgccaga aagaaggact gggaccctaa gaagtacggc
     3901 ggcttcgaca gccccaccgt ggcctattct gtgctggtgg tggccaaagt ggaaaagggc
     3961 aagtccaaga aactgaagag tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc
     4021 agcttcgaga agaatcccat cgactttctg gaagccaagg gctacaaaga agtgaaaaag
     4081 gacctgatca tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg ccggaagaga
     4141 atgctggcct ctgccggcga actgcagaag ggaaacgaac tggccctgcc ctccaaatat
     4201 gtgaacttcc tgtacctggc cagccactat gagaagctga agggctcccc cgaggataat
     4261 gagcagaaac agctgtttgt ggaacagcac aagcactacc tggacgagat catcgagcag
     4321 atcagcgagt tctccaagag agtgatcctg gccgacgcta atctggacaa agtgctgtcc
     4381 gcctacaaca agcaccggga taagcccatc agagagcagg ccgagaatat catccacctg
     4441 tttaccctga ccaatctggg agcccctgcc gccttcaagt actttgacac caccatcgac
     4501 cggaagaggt acaccagcac caaagaggtg ctggacgcca ccctgatcca ccagagcatc
     4561 accggcctgt acgagacacg gatcgacctg tctcagctgg gaggcgacaa aaggccggcg
     4621 gccacgaaaa aggccggcca ggcaaaaaag aaaaagtaag aattcctaga gctcgctgat
     4681 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     4741 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     4801 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     4861 gggaggattg ggaagagaat agcaggcatg ctggggagcg gccgcaggaa cccctagtga
     4921 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     4981 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     5041 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     5101 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     5161 ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt
     5221 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     5281 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     5341 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     5401 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc
     5461 gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga
     5521 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     5581 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     5641 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     5701 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     5761 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     5821 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     5881 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     5941 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     6001 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     6061 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     6121 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     6181 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     6241 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     6301 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     6361 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     6421 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     6481 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     6541 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     6601 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     6661 tggagccggt gagcgtggaa gccgcggtat cattgcagca ctggggccag atggtaagcc
     6721 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     6781 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     6841 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     6901 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     6961 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     7021 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     7081 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     7141 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     7201 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     7261 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     7321 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     7381 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     7441 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     7501 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     7561 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     7621 ttgctggcct tttgctcaca tgtgagggcc tatttcccat gattccttca tatttgcata
     7681 tacgatacaa ggctgttaga gagataattg gaattaattt gactgtaaac acaaagatat
     7741 tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat
     7801 tatgttttaa aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg
     7861 ctttatatat cttgtggaaa ggacgaaaca ccggagacct gtacaggtct ctgttttaga
     7921 gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga
     7981 gtcggtgctt ttttgtttta gagctagaaa tagcaagtta aaataaggct agtccgtttt
     8041 tagcgcgtgc gccaattctg cagacaaatg gctctagagg tacccgttac ataacttacg
     8101 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aatagtaacg
     8161 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     8221 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     8281 tggcccgcct ggcattgtgc ccagtacatg accttatggg actttcctac ttggcagtac
     8341 atctacgtat tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact
     8401 ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt
     8461 tgtgcagcga tgggggcggg gggggggggg gggcggcscc aggcgggggg gggggggggg
     8521 gggggggggg gggggggggg gggggggggg gggggcgcgc gccaggcgg
//
