LOCUS       sequence-334778-        8947 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Cas9 and 2 guides.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8947)
  AUTHORS   Fang H, Bygrave AM, Roth RH, Johnson RC, Huganir RL
  TITLE     An optimized CRISPR/Cas9 approach for precise genome editing in 
            neurons.
  JOURNAL   Elife. 2021 Mar 10;10. pii: 65202. doi: 10.7554/eLife.65202.
  PUBMED    33689678
REFERENCE   2  (bases 1 to 8947)
  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: "Elife."; 
            date: "2021-03-10"; volume: "10. pii: 65202. doi"; pages: " 
            10.7554/eLife.65202"
COMMENT     Sequence Label: rGluA2 (px330:2-guideCas9)
FEATURES             Location/Qualifiers
     source          1..8947
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          110..337
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      349..358
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             358..423
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             430..450
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             475..4575
                     /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             475..4575
                     /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             4576..4623
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(4651..4668)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4657..4864
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4873..5013
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4873..5002
                     /label=AAV2 ITR
     rep_origin      5088..5543
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5175..5194)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5385..5406
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5560..5579)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5679..5701
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5739..5757)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5825..5929
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5930..5998,5999..6790)
                     /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(6148..6167)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6961..7549
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7450..7469
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        7611..7859
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        7881..7956
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        8047..8295
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        8317..8392
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8492..8777
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 gsggrgaggk gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc
       61 gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc
      121 gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc cccggctctg
      181 actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa
      241 ttagctgagc aagaggtaag ggtttaaggg atggttggtt ggtggggtat taatgtttaa
      301 ttacctggag cacctgcctg aaatcacttt ttttcaggtt ggaccggtgc caccatggac
      361 tataaggacc acgacggaga ctacaaggat catgatattg attacaaaga cgatgacgat
      421 aagatggccc caaagaagaa gcggaaggtc ggtatccacg gagtcccagc agccgacaag
      481 aagtacagca tcggcctgga catcggcacc aactctgtgg gctgggccgt gatcaccgac
      541 gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc
      601 aagaagaacc tgatcggagc cctgctgttc gacagcggcg aaacagccga ggccacccgg
      661 ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa
      721 gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag
      781 tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg
      841 gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg
      901 gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag
      961 ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag
     1021 ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc
     1081 agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa
     1141 aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggaaa cctgattgcc
     1201 ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa
     1261 ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc
     1321 gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc
     1381 gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag
     1441 agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg
     1501 cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatt
     1561 gacggcggag ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg
     1621 gacggcaccg aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg
     1681 accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg
     1741 cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc
     1801 ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc
     1861 tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac
     1921 aagggcgctt ccgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc
     1981 aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtataacgag
     2041 ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag
     2101 cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag
     2161 ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg
     2221 gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac
     2281 aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg
     2341 acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc
     2401 gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc
     2461 cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg
     2521 aagtccgacg gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc
     2581 tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac
     2641 attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg
     2701 gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg
     2761 gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg
     2821 atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac
     2881 acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac
     2941 gtggaccagg aactggacat caaccggctg tccgactacg atgtggacca tatcgtgcct
     3001 cagagctttc tgaaggacga ctccatcgac aacaaggtgc tgaccagaag cgacaagaac
     3061 cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg
     3121 cggcagctgc tgaacgccaa gctgattacc cagagaaagt tcgacaatct gaccaaggcc
     3181 gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa
     3241 acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac
     3301 gacgagaatg acaagctgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg
     3361 tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac
     3421 gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag
     3481 ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc
     3541 aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg
     3601 aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc
     3661 gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg gccgggattt tgccaccgtg
     3721 cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc
     3781 ggcttcagca aagagtctat cctgcccaag aggaacagcg ataagctgat cgccagaaag
     3841 aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg
     3901 ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg
     3961 ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa
     4021 gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg
     4081 ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga
     4141 aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag
     4201 aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaag
     4261 cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc
     4321 gacgctaatc tggacaaagt gctgtccgcc tacaacaagc accgggataa gcccatcaga
     4381 gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc
     4441 ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg
     4501 gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct
     4561 cagctgggag gcgacaaaag gccggcggcc acgaaaaagg ccggccaggc aaaaaagaaa
     4621 aagtaagaat tcctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat
     4681 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     4741 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     4801 ggggtggggt ggggcaggac agcaaggggg aggattggga agagaatagc aggcatgctg
     4861 gggagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     4921 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     4981 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     5041 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     5101 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     5161 cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     5221 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     5281 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     5341 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     5401 aactggaaca acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc
     5461 gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     5521 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     5581 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     5641 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     5701 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     5761 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     5821 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     5881 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     5941 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     6001 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     6061 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     6121 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     6181 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     6241 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     6301 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     6361 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     6421 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     6481 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     6541 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     6601 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtggaagcc gcggtatcat
     6661 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     6721 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     6781 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     6841 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     6901 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     6961 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     7021 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     7081 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     7141 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     7201 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     7261 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     7321 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     7381 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     7441 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     7501 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     7561 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt gagggcctat
     7621 ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag ataattggaa
     7681 ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat
     7741 ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgcttaccg
     7801 taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga cgaaacaccg
     7861 atatcgacct cacaatgcag gttttagagc tagaaatagc aagttaaaat aaggctagtc
     7921 cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgttttaga gctagaaata
     7981 gcaagttaaa ataaggctag tccgttttta gcgcgtgcgc caattctgca gacaaatggc
     8041 tctagcgagg gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt
     8101 agagagataa ttggaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga
     8161 cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac
     8221 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg
     8281 aaaggacgaa acaccgtatg ccttttgaca caataggttt tagagctaga aatagcaagt
     8341 taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gcttttttgt
     8401 tttagagcta gaaatagcaa gttaaaataa ggctagtccg tttttagcgc gtgcgccaat
     8461 tctgcagaca aatggctcta gaggtggtac ccgttacata acttacggta aatggcccgc
     8521 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat agtaacgcca atagggactt
     8581 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     8641 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     8701 attgtgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     8761 tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc
     8821 ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg
     8881 gggcgggggg gggggggggg cgcgcgccrg gsggggsggg gsggggsgrg gggsggggsg
     8941 gggsgrg
//
