LOCUS       sequence-334742-        8666 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used for guide cloning with PCR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8666)
  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 8666)
  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: pX52
FEATURES             Location/Qualifiers
     source          1..8666
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          126..353
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      365..374
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             374..439
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             446..466
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             491..4591
                     /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             491..4591
                     /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             4592..4639
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(4667..4684)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4673..4880
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4889..5029
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4889..5018
                     /label=AAV2 ITR
     rep_origin      5104..5559
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5191..5210)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5401..5422
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5576..5595)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5695..5717
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5755..5773)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5841..5945
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5946..6014,6015..6806)
                     /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(6164..6183)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6977..7565
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7466..7485
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        7627..7875
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        7894..7969
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        8008..8256
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     enhancer        8264..8549
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa
       61 agtttccttt tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc
      121 gggcgggagt cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc
      181 gcccgccccg gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt
      241 ctcctccggg ctgtaattag ctgagcaaga ggtaagggtt taagggatgg ttggttggtg
      301 gggtattaat gtttaattac ctggagcacc tgcctgaaat cacttttttt caggttggac
      361 cggtgccacc atggactata aggaccacga cggagactac aaggatcatg atattgatta
      421 caaagacgat gacgataaga tggccccaaa gaagaagcgg aaggtcggta tccacggagt
      481 cccagcagcc gacaagaagt acagcatcgg cctggacatc ggcaccaact ctgtgggctg
      541 ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac
      601 cgaccggcac agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac
      661 agccgaggcc acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg
      721 gatctgctat ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt
      781 ccacagactg gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat
      841 cttcggcaac atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct
      901 gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct
      961 ggcccacatg atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa
     1021 cagcgacgtg gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga
     1081 aaaccccatc aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa
     1141 gagcagacgg ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt
     1201 cggaaacctg attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct
     1261 ggccgaggat gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct
     1321 gctggcccag atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga
     1381 cgccatcctg ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag
     1441 cgcctctatg atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct
     1501 cgtgcggcag cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg
     1561 ctacgccggc tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc
     1621 catcctggaa aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct
     1681 gctgcggaag cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga
     1741 gctgcacgcc attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga
     1801 aaagatcgag aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg
     1861 aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt
     1921 cgaggaagtg gtggacaagg gcgcttccgc ccagagcttc atcgagcgga tgaccaactt
     1981 cgataagaac ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt
     2041 caccgtgtat aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc
     2101 cttcctgagc ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa
     2161 agtgaccgtg aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt
     2221 ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct
     2281 gaaaattatc aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga
     2341 tatcgtgctg accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac
     2401 ctatgcccac ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg
     2461 ctggggcagg ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac
     2521 aatcctggat ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca
     2581 cgacgacagc ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga
     2641 tagcctgcac gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct
     2701 gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa
     2761 catcgtgatc gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg
     2821 cgagagaatg aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga
     2881 acaccccgtg gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa
     2941 tgggcgggat atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt
     3001 ggaccatatc gtgcctcaga gctttctgaa ggacgactcc atcgacaaca aggtgctgac
     3061 cagaagcgac aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa
     3121 gatgaagaac tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga
     3181 caatctgacc aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa
     3241 gagacagctg gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg
     3301 gatgaacact aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct
     3361 gaagtccaag ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat
     3421 caacaactac caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat
     3481 caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt
     3541 gcggaagatg atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt
     3601 ctacagcaac atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg
     3661 gaagcggcct ctgatcgaga caaacggcga aaccggggag atcgtgtggg ataagggccg
     3721 ggattttgcc accgtgcgga aagtgctgag catgccccaa gtgaatatcg tgaaaaagac
     3781 cgaggtgcag acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgataa
     3841 gctgatcgcc agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac
     3901 cgtggcctat tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa
     3961 gagtgtgaaa gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc
     4021 catcgacttt ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct
     4081 gcctaagtac tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg
     4141 cgaactgcag aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct
     4201 ggccagccac tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt
     4261 tgtggaacag cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa
     4321 gagagtgatc ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg
     4381 ggataagccc atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct
     4441 gggagcccct gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag
     4501 caccaaagag gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac
     4561 acggatcgac ctgtctcagc tgggaggcga caaaaggccg gcggccacga aaaaggccgg
     4621 ccaggcaaaa aagaaaaagt aagaattcct agagctcgct gatcagcctc gactgtgcct
     4681 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     4741 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     4801 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagag
     4861 aatagcaggc atgctgggga gcggccgcag gaacccctag tgatggagtt ggccactccc
     4921 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     4981 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
     5041 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     5101 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     5161 ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg
     5221 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     5281 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     5341 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     5401 gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt
     5461 tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat
     5521 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     5581 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     5641 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     5701 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     5761 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     5821 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5881 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5941 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     6001 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     6061 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     6121 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     6181 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     6241 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     6301 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     6361 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     6421 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     6481 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     6541 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     6601 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     6661 gaagccgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     6721 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     6781 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     6841 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6901 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6961 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     7021 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     7081 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     7141 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     7201 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     7261 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     7321 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     7381 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     7441 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     7501 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     7561 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     7621 acatgtgagg gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt
     7681 agagagataa ttggaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga
     7741 cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac
     7801 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg
     7861 aaaggacgaa acaccgggtc ttcgagaaga cctgttttag agctagaaat agcaagttaa
     7921 aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct tttttctaga
     7981 ccaattctgc agacaaatgg ctctagcgag ggcctatttc ccatgattcc ttcatatttg
     8041 catatacgat acaaggctgt tagagagata attggaatta atttgactgt aaacacaaag
     8101 atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta
     8161 aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc
     8221 ttggctttat atatcttgtg gaaaggacga aacaccgggt acccgttaca taacttacgg
     8281 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca atagtaacgc
     8341 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     8401 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     8461 ggcccgcctg gcattgtgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     8521 tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc
     8581 tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt
     8641 gtgcagcgat gggggcgggg gggggg
//
