LOCUS       sequence-334774-        8945 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 8945)
  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 8945)
  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: beta2 (px330:2-guideCas9)
FEATURES             Location/Qualifiers
     source          1..8945
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          123..350
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      362..371
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             371..436
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             443..463
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(488..3025,3026..3028,3029..3490,3491..3493,3494..3661,
                     3662..3664,3665..4588)
                     /codon_start=1
                     /product="Cas9 endonuclease from the Streptococcus pyogenes
                     Type II CRISPR/Cas system, mutated to improve targeting 
                     specificity (Slaymaker et al., 2016)"
                     /label=eSpCas9(1.1)
                     /note="carries the mutations K848A, K1003A, and R1060A"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLADDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPALESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKAPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4589..4636
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(4664..4681)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4670..4877
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4886..5026
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4886..5015
                     /label=AAV2 ITR
     rep_origin      5101..5556
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5188..5207)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5398..5419
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5573..5592)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5692..5714
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5752..5770)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5838..5942
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5943..6011,6012..6803)
                     /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(6161..6180)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6974..7562
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7463..7482
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        7624..7872
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        7893..7968
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        8059..8307
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        8329..8404
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8504..8789
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 gggggggggs grggsggrga ggkgcggcgg cagccaatca gagcggcgcg ctccgaaagt
       61 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      121 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      181 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      241 ctccgggctg taattagctg agcaagaggt aagggtttaa gggatggttg gttggtgggg
      301 tattaatgtt taattacctg gagcacctgc ctgaaatcac tttttttcag gttggaccgg
      361 tgccaccatg gactataagg accacgacgg agactacaag gatcatgata ttgattacaa
      421 agacgatgac gataagatgg ccccaaagaa gaagcggaag gtcggtatcc acggagtccc
      481 agcagccgac aagaagtaca gcatcggcct ggacatcggc accaactctg tgggctgggc
      541 cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga
      601 ccggcacagc atcaagaaga acctgatcgg agccctgctg ttcgacagcg gcgaaacagc
      661 cgaggccacc cggctgaaga gaaccgccag aagaagatac accagacgga agaaccggat
      721 ctgctatctg caagagatct tcagcaacga gatggccaag gtggacgaca gcttcttcca
      781 cagactggaa gagtccttcc tggtggaaga ggataagaag cacgagcggc accccatctt
      841 cggcaacatc gtggacgagg tggcctacca cgagaagtac cccaccatct accacctgag
      901 aaagaaactg gtggacagca ccgacaaggc cgacctgcgg ctgatctatc tggccctggc
      961 ccacatgatc aagttccggg gccacttcct gatcgagggc gacctgaacc ccgacaacag
     1021 cgacgtggac aagctgttca tccagctggt gcagacctac aaccagctgt tcgaggaaaa
     1081 ccccatcaac gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag
     1141 cagacggctg gaaaatctga tcgcccagct gcccggcgag aagaagaatg gcctgttcgg
     1201 aaacctgatt gccctgagcc tgggcctgac ccccaacttc aagagcaact tcgacctggc
     1261 cgaggatgcc aaactgcagc tgagcaagga cacctacgac gacgacctgg acaacctgct
     1321 ggcccagatc ggcgaccagt acgccgacct gtttctggcc gccaagaacc tgtccgacgc
     1381 catcctgctg agcgacatcc tgagagtgaa caccgagatc accaaggccc ccctgagcgc
     1441 ctctatgatc aagagatacg acgagcacca ccaggacctg accctgctga aagctctcgt
     1501 gcggcagcag ctgcctgaga agtacaaaga gattttcttc gaccagagca agaacggcta
     1561 cgccggctac attgacggcg gagccagcca ggaagagttc tacaagttca tcaagcccat
     1621 cctggaaaag atggacggca ccgaggaact gctcgtgaag ctgaacagag aggacctgct
     1681 gcggaagcag cggaccttcg acaacggcag catcccccac cagatccacc tgggagagct
     1741 gcacgccatt ctgcggcggc aggaagattt ttacccattc ctgaaggaca accgggaaaa
     1801 gatcgagaag atcctgacct tccgcatccc ctactacgtg ggccctctgg ccaggggaaa
     1861 cagcagattc gcctggatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga
     1921 ggaagtggtg gacaagggcg cttccgccca gagcttcatc gagcggatga ccaacttcga
     1981 taagaacctg cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac
     2041 cgtgtataac gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt
     2101 cctgagcggc gagcagaaaa aggccatcgt ggacctgctg ttcaagacca accggaaagt
     2161 gaccgtgaag cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga
     2221 aatctccggc gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa
     2281 aattatcaag gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat
     2341 cgtgctgacc ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta
     2401 tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag cggcggagat acaccggctg
     2461 gggcaggctg agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat
     2521 cctggatttc ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga
     2581 cgacagcctg acctttaaag aggacatcca gaaagcccag gtgtccggcc agggcgatag
     2641 cctgcacgag cacattgcca atctggccgg cagccccgcc attaagaagg gcatcctgca
     2701 gacagtgaag gtggtggacg agctcgtgaa agtgatgggc cggcacaagc ccgagaacat
     2761 cgtgatcgaa atggccagag agaaccagac cacccagaag ggacagaaga acagccgcga
     2821 gagaatgaag cggatcgaag agggcatcaa agagctgggc agccagatcc tgaaagaaca
     2881 ccccgtggaa aacacccagc tgcagaacga gaagctgtac ctgtactacc tgcagaatgg
     2941 gcgggatatg tacgtggacc aggaactgga catcaaccgg ctgtccgact acgatgtgga
     3001 ccatatcgtg cctcagagct ttctggccga cgactccatc gacaacaagg tgctgaccag
     3061 aagcgacaag aaccggggca agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat
     3121 gaagaactac tggcggcagc tgctgaacgc caagctgatt acccagagaa agttcgacaa
     3181 tctgaccaag gccgagagag gcggcctgag cgaactggat aaggccggct tcatcaagag
     3241 acagctggtg gaaacccggc agatcacaaa gcacgtggca cagatcctgg actcccggat
     3301 gaacactaag tacgacgaga atgacaagct gatccgggaa gtgaaagtga tcaccctgaa
     3361 gtccaagctg gtgtccgatt tccggaagga tttccagttt tacaaagtgc gcgagatcaa
     3421 caactaccac cacgcccacg acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa
     3481 aaagtaccct gcgctggaaa gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg
     3541 gaagatgatc gccaagagcg agcaggaaat cggcaaggct accgccaagt acttcttcta
     3601 cagcaacatc atgaactttt tcaagaccga gattaccctg gccaacggcg agatccggaa
     3661 ggcgcctctg atcgagacaa acggcgaaac cggggagatc gtgtgggata agggccggga
     3721 ttttgccacc gtgcggaaag tgctgagcat gccccaagtg aatatcgtga aaaagaccga
     3781 ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc aagaggaaca gcgataagct
     3841 gatcgccaga aagaaggact gggaccctaa gaagtacggc ggcttcgaca gccccaccgt
     3901 ggcctattct gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga aactgaagag
     3961 tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc agcttcgaga agaatcccat
     4021 cgactttctg gaagccaagg gctacaaaga agtgaaaaag gacctgatca tcaagctgcc
     4081 taagtactcc ctgttcgagc tggaaaacgg ccggaagaga atgctggcct ctgccggcga
     4141 actgcagaag ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc tgtacctggc
     4201 cagccactat gagaagctga agggctcccc cgaggataat gagcagaaac agctgtttgt
     4261 ggaacagcac aagcactacc tggacgagat catcgagcag atcagcgagt tctccaagag
     4321 agtgatcctg gccgacgcta atctggacaa agtgctgtcc gcctacaaca agcaccggga
     4381 taagcccatc agagagcagg ccgagaatat catccacctg tttaccctga ccaatctggg
     4441 agcccctgcc gccttcaagt actttgacac caccatcgac cggaagaggt acaccagcac
     4501 caaagaggtg ctggacgcca ccctgatcca ccagagcatc accggcctgt acgagacacg
     4561 gatcgacctg tctcagctgg gaggcgacaa aaggccggcg gccacgaaaa aggccggcca
     4621 ggcaaaaaag aaaaagtaag aattcctaga gctcgctgat cagcctcgac tgtgccttct
     4681 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     4741 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     4801 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagagaat
     4861 agcaggcatg ctggggagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     4921 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     4981 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     5041 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     5101 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     5161 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     5221 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     5281 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     5341 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     5401 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     5461 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     5521 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     5581 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     5641 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     5701 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     5761 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     5821 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     5881 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     5941 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     6001 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     6061 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     6121 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     6181 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     6241 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     6301 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     6361 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     6421 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     6481 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     6541 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     6601 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtggaa
     6661 gccgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     6721 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     6781 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     6841 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     6901 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     6961 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     7021 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     7081 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     7141 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     7201 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     7261 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     7321 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     7381 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     7441 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     7501 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     7561 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     7621 tgtgagggcc tatttcccat gattccttca tatttgcata tacgatacaa ggctgttaga
     7681 gagataattg gaattaattt gactgtaaac acaaagatat tagtacaaaa tacgtgacgt
     7741 agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa aatggactat
     7801 catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat cttgtggaaa
     7861 ggacgaaaca ccggcgtacc aaaacatcaa aggttttaga gctagaaata gcaagttaaa
     7921 ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt ttttgtttta
     7981 gagctagaaa tagcaagtta aaataaggct agtccgtttt tagcgcgtgc gccaattctg
     8041 cagacaaatg gctctagcga gggcctattt cccatgattc cttcatattt gcatatacga
     8101 tacaaggctg ttagagagat aattggaatt aatttgactg taaacacaaa gatattagta
     8161 caaaatacgt gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt
     8221 tttaaaatgg actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta
     8281 tatatcttgt ggaaaggacg aaacaccgag tctggtcaca ctctaagggt tttagagcta
     8341 gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg
     8401 gtgctttttt gttttagagc tagaaatagc aagttaaaat aaggctagtc cgtttttagc
     8461 gcgtgcgcca attctgcaga caaatggctc tagaggtggt acccgttaca taacttacgg
     8521 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca atagtaacgc
     8581 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     8641 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     8701 ggcccgcctg gcattgtgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     8761 tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc
     8821 tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt
     8881 gtgcagcgat gggggcgggg gggggggggg ggcgcgcgcc gggggggggg gggggggggg
     8941 ggggg
//
