LOCUS       sequence-335498-        8946 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 8946)
  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 8946)
  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: GluN1 (px330:2-guideCas9)
FEATURES             Location/Qualifiers
     source          1..8946
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        270..345
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        436..684
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        706..781
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        881..1166
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        1168..1445
                     /label=chicken beta-actin promoter
     intron          1446..1673
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1685..1694
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1694..1759
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1766..1786
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1811..5911
                     /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             1811..5911
                     /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             5912..5959
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(5987..6004)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5993..6200
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6209..6349
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6209..6338
                     /label=AAV2 ITR
     rep_origin      6424..6879
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6511..6530)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6721..6742
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6896..6915)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7015..7037
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7075..7093)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7161..7265
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7266..7334,7335..8126)
                     /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(7484..7503)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8297..8885
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8786..8805
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg cgctgctcga acacccgcgg ttttagagct agaaatagca agttaaaata
      301 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt tgttttagag
      361 ctagaaatag caagttaaaa taaggctagt ccgtttttag cgcgtgcgcc aattctgcag
      421 acaaatggct ctagcgaggg cctatttccc atgattcctt catatttgca tatacgatac
      481 aaggctgtta gagagataat tggaattaat ttgactgtaa acacaaagat attagtacaa
      541 aatacgtgac gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt
      601 aaaatggact atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat
      661 atcttgtgga aaggacgaaa caccgtctgt tccatctagt cagtggtttt agagctagaa
      721 atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg
      781 cttttttgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt ttttagcgcg
      841 tgcgccaatt ctgcagacaa atggctctag aggtggtacc cgttacataa cttacggtaa
      901 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata gtaacgccaa
      961 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     1021 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     1081 ccgcctggca ttgtgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     1141 acgtattagt catcgctatt accatggtcg aggtgagccc cacgttctgc ttcactctcc
     1201 ccatctcccc cccctcccca cccccaattt tgtatttatt tattttttaa ttattttgtg
     1261 cagcgatggg ggcggggggg gggggggggc gcgcgccrgg sggggsgggg sggggsgrgg
     1321 ggsggggsgg ggsgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa
     1381 agtttccttt tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc
     1441 gggcgggagt cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc
     1501 gcccgccccg gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt
     1561 ctcctccggg ctgtaattag ctgagcaaga ggtaagggtt taagggatgg ttggttggtg
     1621 gggtattaat gtttaattac ctggagcacc tgcctgaaat cacttttttt caggttggac
     1681 cggtgccacc atggactata aggaccacga cggagactac aaggatcatg atattgatta
     1741 caaagacgat gacgataaga tggccccaaa gaagaagcgg aaggtcggta tccacggagt
     1801 cccagcagcc gacaagaagt acagcatcgg cctggacatc ggcaccaact ctgtgggctg
     1861 ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac
     1921 cgaccggcac agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac
     1981 agccgaggcc acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg
     2041 gatctgctat ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt
     2101 ccacagactg gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat
     2161 cttcggcaac atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct
     2221 gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct
     2281 ggcccacatg atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa
     2341 cagcgacgtg gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga
     2401 aaaccccatc aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa
     2461 gagcagacgg ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt
     2521 cggaaacctg attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct
     2581 ggccgaggat gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct
     2641 gctggcccag atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga
     2701 cgccatcctg ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag
     2761 cgcctctatg atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct
     2821 cgtgcggcag cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg
     2881 ctacgccggc tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc
     2941 catcctggaa aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct
     3001 gctgcggaag cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga
     3061 gctgcacgcc attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga
     3121 aaagatcgag aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg
     3181 aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt
     3241 cgaggaagtg gtggacaagg gcgcttccgc ccagagcttc atcgagcgga tgaccaactt
     3301 cgataagaac ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt
     3361 caccgtgtat aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc
     3421 cttcctgagc ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa
     3481 agtgaccgtg aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt
     3541 ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct
     3601 gaaaattatc aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga
     3661 tatcgtgctg accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac
     3721 ctatgcccac ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg
     3781 ctggggcagg ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac
     3841 aatcctggat ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca
     3901 cgacgacagc ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga
     3961 tagcctgcac gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct
     4021 gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa
     4081 catcgtgatc gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg
     4141 cgagagaatg aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga
     4201 acaccccgtg gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa
     4261 tgggcgggat atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt
     4321 ggaccatatc gtgcctcaga gctttctgaa ggacgactcc atcgacaaca aggtgctgac
     4381 cagaagcgac aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa
     4441 gatgaagaac tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga
     4501 caatctgacc aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa
     4561 gagacagctg gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg
     4621 gatgaacact aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct
     4681 gaagtccaag ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat
     4741 caacaactac caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat
     4801 caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt
     4861 gcggaagatg atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt
     4921 ctacagcaac atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg
     4981 gaagcggcct ctgatcgaga caaacggcga aaccggggag atcgtgtggg ataagggccg
     5041 ggattttgcc accgtgcgga aagtgctgag catgccccaa gtgaatatcg tgaaaaagac
     5101 cgaggtgcag acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgataa
     5161 gctgatcgcc agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac
     5221 cgtggcctat tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa
     5281 gagtgtgaaa gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc
     5341 catcgacttt ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct
     5401 gcctaagtac tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg
     5461 cgaactgcag aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct
     5521 ggccagccac tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt
     5581 tgtggaacag cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa
     5641 gagagtgatc ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg
     5701 ggataagccc atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct
     5761 gggagcccct gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag
     5821 caccaaagag gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac
     5881 acggatcgac ctgtctcagc tgggaggcga caaaaggccg gcggccacga aaaaggccgg
     5941 ccaggcaaaa aagaaaaagt aagaattcct agagctcgct gatcagcctc gactgtgcct
     6001 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     6061 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     6121 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagag
     6181 aatagcaggc atgctgggga gcggccgcag gaacccctag tgatggagtt ggccactccc
     6241 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     6301 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
     6361 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     6421 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     6481 ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg
     6541 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     6601 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     6661 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     6721 gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt
     6781 tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat
     6841 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     6901 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     6961 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     7021 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     7081 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     7141 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     7201 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     7261 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     7321 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     7381 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     7441 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     7501 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     7561 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     7621 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     7681 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     7741 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     7801 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     7861 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     7921 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     7981 gaagccgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     8041 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     8101 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     8161 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     8221 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     8281 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     8341 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     8401 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     8461 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     8521 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     8581 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     8641 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     8701 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     8761 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     8821 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     8881 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     8941 acatgt
//
