LOCUS       sequence-335129-        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: GluA3 (px330:2-guideCas9)
FEATURES             Location/Qualifiers
     source          1..8946
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          163..390
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      402..411
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             411..476
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             483..503
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             528..4628
                     /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             528..4628
                     /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             4629..4676
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(4704..4721)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4710..4917
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4926..5066
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4926..5055
                     /label=AAV2 ITR
     rep_origin      5141..5596
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5228..5247)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5438..5459
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5613..5632)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5732..5754
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5792..5810)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5878..5982
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5983..6051,6052..6843)
                     /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(6201..6220)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7014..7602
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7503..7522
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        7664..7912
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        7933..8008
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        8099..8347
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        8369..8444
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8544..8829
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        join(8831..8946,1..162)
                     /label=chicken beta-actin promoter
ORIGIN
        1 ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc ggggcggggc gaggcggaga
       61 ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg
      121 cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc
      181 cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc
      241 gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg taattagctg
      301 agcaagaggt aagggtttaa gggatggttg gttggtgggg tattaatgtt taattacctg
      361 gagcacctgc ctgaaatcac tttttttcag gttggaccgg tgccaccatg gactataagg
      421 accacgacgg agactacaag gatcatgata ttgattacaa agacgatgac gataagatgg
      481 ccccaaagaa gaagcggaag gtcggtatcc acggagtccc agcagccgac aagaagtaca
      541 gcatcggcct ggacatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca
      601 aggtgcccag caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga
      661 acctgatcgg agccctgctg ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga
      721 gaaccgccag aagaagatac accagacgga agaaccggat ctgctatctg caagagatct
      781 tcagcaacga gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc
      841 tggtggaaga ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg
      901 tggcctacca cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca
      961 ccgacaaggc cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg
     1021 gccacttcct gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca
     1081 tccagctggt gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg
     1141 tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga
     1201 tcgcccagct gcccggcgag aagaagaatg gcctgttcgg aaacctgatt gccctgagcc
     1261 tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc
     1321 tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt
     1381 acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc
     1441 tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg
     1501 acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga
     1561 agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac attgacggcg
     1621 gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca
     1681 ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg
     1741 acaacggcag catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc
     1801 aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct
     1861 tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga
     1921 ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg
     1981 cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga
     2041 aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac gagctgacca
     2101 aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa
     2161 aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag
     2221 aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc
     2281 ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact
     2341 tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt
     2401 ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca
     2461 aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc
     2521 tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg
     2581 acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag
     2641 aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca
     2701 atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg
     2761 agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag
     2821 agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag
     2881 agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc
     2941 tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc
     3001 aggaactgga catcaaccgg ctgtccgact acgatgtgga ccatatcgtg cctcagagct
     3061 ttctgaagga cgactccatc gacaacaagg tgctgaccag aagcgacaag aaccggggca
     3121 agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcggcagc
     3181 tgctgaacgc caagctgatt acccagagaa agttcgacaa tctgaccaag gccgagagag
     3241 gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc
     3301 agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga
     3361 atgacaagct gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt
     3421 tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg
     3481 acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa
     3541 gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg
     3601 agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt
     3661 tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa
     3721 acggcgaaac cggggagatc gtgtgggata agggccggga ttttgccacc gtgcggaaag
     3781 tgctgagcat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca
     3841 gcaaagagtc tatcctgccc aagaggaaca gcgataagct gatcgccaga aagaaggact
     3901 gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct gtgctggtgg
     3961 tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctgctgggga
     4021 tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg
     4081 gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc
     4141 tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag ggaaacgaac
     4201 tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga
     4261 agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac aagcactacc
     4321 tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta
     4381 atctggacaa agtgctgtcc gcctacaaca agcaccggga taagcccatc agagagcagg
     4441 ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt
     4501 actttgacac caccatcgac cggaagaggt acaccagcac caaagaggtg ctggacgcca
     4561 ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg
     4621 gaggcgacaa aaggccggcg gccacgaaaa aggccggcca ggcaaaaaag aaaaagtaag
     4681 aattcctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     4741 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     4801 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     4861 ggtggggcag gacagcaagg gggaggattg ggaagagaat agcaggcatg ctggggagcg
     4921 gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     4981 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     5041 cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct
     5101 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
     5161 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta
     5221 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     5281 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     5341 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     5401 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     5461 acaacactca actctatctc gggctattct tttgatttat aagggatttt gccgatttcg
     5521 gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     5581 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     5641 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     5701 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     5761 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     5821 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     5881 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     5941 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     6001 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     6061 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     6121 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     6181 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     6241 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     6301 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     6361 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     6421 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     6481 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     6541 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     6601 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     6661 tggtttattg ctgataaatc tggagccggt gagcgtggaa gccgcggtat cattgcagca
     6721 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     6781 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     6841 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     6901 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     6961 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     7021 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     7081 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     7141 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     7201 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     7261 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     7321 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     7381 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     7441 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     7501 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     7561 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     7621 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtgagggcc tatttcccat
     7681 gattccttca tatttgcata tacgatacaa ggctgttaga gagataattg gaattaattt
     7741 gactgtaaac acaaagatat tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg
     7801 gtagtttgca gttttaaaat tatgttttaa aatggactat catatgctta ccgtaacttg
     7861 aaagtatttc gatttcttgg ctttatatat cttgtggaaa ggacgaaaca ccgcgagcga
     7921 gagcaagttg aggttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca
     7981 acttgaaaaa gtggcaccga gtcggtgctt ttttgtttta gagctagaaa tagcaagtta
     8041 aaataaggct agtccgtttt tagcgcgtgc gccaattctg cagacaaatg gctctagcga
     8101 gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg ttagagagat
     8161 aattggaatt aatttgactg taaacacaaa gatattagta caaaatacgt gacgtagaaa
     8221 gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg actatcatat
     8281 gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt ggaaaggacg
     8341 aaacaccgct gcaagaggct aagagtcggt tttagagcta gaaatagcaa gttaaaataa
     8401 ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt gttttagagc
     8461 tagaaatagc aagttaaaat aaggctagtc cgtttttagc gcgtgcgcca attctgcaga
     8521 caaatggctc tagaggtggt acccgttaca taacttacgg taaatggccc gcctggctga
     8581 ccgcccaacg acccccgccc attgacgtca atagtaacgc caatagggac tttccattga
     8641 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     8701 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattgtgcc
     8761 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     8821 attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc
     8881 ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg
     8941 gggggg
//
