LOCUS       sequence-363164-        8609 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of spCas9, gRNA targeting the end of mouse Nefl gene and 
            donor linker(A6TAG)-3xFLAG. Can be used for amber codon suppression 
            and click chemistry labeling of endogenous NFL..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8609)
  AUTHORS   Arsic A, Hagemann C, Stajkovic N, Schubert T, Nikic-Spiegel I
  TITLE     Minimal genetically encoded tags for fluorescent protein labeling in
            living neurons.
  JOURNAL   Nat Commun. 2022 Jan 14;13(1):314. doi: 10.1038/s41467-022-27956-y.
  PUBMED    35031604
REFERENCE   2  (bases 1 to 8609)
  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: "Nat 
            Commun."; date: "2022-01-14"; volume: "13(1):314. doi"; pages: " 
            10.1038/s41467-022-27956-y"
COMMENT     Sequence Label: pORANGE_NFL linker(A6TAG)-3xFLAG KI
FEATURES             Location/Qualifiers
     source          1..8609
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     130..149
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        228..303
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             440..505
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     enhancer        571..856
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        858..1135
                     /label=chicken beta-actin promoter
     intron          1136..1363
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             1384..1410
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1414..1434
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1444..5544
                     /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             1444..5544
                     /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             5548..5568
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(5608..5625)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5614..5821
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5830..5970
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5830..5959
                     /label=AAV2 ITR
     rep_origin      6045..6500
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6132..6151)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6342..6363
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6517..6536)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6636..6658
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6696..6714)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6782..6886
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6887..6955,6956..7747)
                     /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(7105..7124)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7918..8506
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8407..8426
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(8568..8609,1..207)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8568..8588
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 tacaaggctg ttagagagat aattggaatt aatttgactg taaacacaaa gatattagta
       61 caaaatacgt gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt
      121 tttaaaatgg actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta
      181 tatatcttgt ggaaaggacg aaacaccgag tgctggagag gagcagggtt ttagagctag
      241 aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg
      301 tgcttttttg ttttagagct agaaatagca agttaaaata aggctagtcc gtttttagcg
      361 cgtgcgccaa ttctgcagac aaatggctct agaagcttcc acctgctcct ctccagcact
      421 cggtagcgct ggaagctagg actacaaaga ccatgacggt gattataaag atcatgacat
      481 cgattacaag gatgacgatg acaagtaacc acctgctcct ctccagcact cggatcctag
      541 gacgcgtcct aggacgcgtc cagaggtacc cgttacataa cttacggtaa atggcccgcc
      601 tggctgaccg cccaacgacc cccgcccatt gacgtcaata gtaacgccaa tagggacttt
      661 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      721 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      781 ttgtgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      841 catcgctatt accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc
      901 cccctcccca cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg
      961 ggcggggggg gggggggggc gcgcgccggg sggggcgggg cggggcgagg ggcggggcgg
     1021 ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt
     1081 tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt
     1141 cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg
     1201 gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg
     1261 ctgtaattag ctgagcaaga ggtaagggtt taagggatgg ttggttggtg gggtattaat
     1321 gtttaattac ctggagcacc tgcctgaaat cacttttttt caggttggac cggtgccacc
     1381 atgtacccat acgatgttcc agattacgct tcgccgaaga aaaagcgcaa ggtcgaagcg
     1441 tccgacaaga agtacagcat cggcctggac atcggcacca actctgtggg ctgggccgtg
     1501 atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa caccgaccgg
     1561 cacagcatca agaagaacct gatcggagcc ctgctgttcg acagcggcga aacagccgag
     1621 gccacccggc tgaagagaac cgccagaaga agatacacca gacggaagaa ccggatctgc
     1681 tatctgcaag agatcttcag caacgagatg gccaaggtgg acgacagctt cttccacaga
     1741 ctggaagagt ccttcctggt ggaagaggat aagaagcacg agcggcaccc catcttcggc
     1801 aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca cctgagaaag
     1861 aaactggtgg acagcaccga caaggccgac ctgcggctga tctatctggc cctggcccac
     1921 atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac
     1981 gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga ggaaaacccc
     2041 atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg ccagactgag caagagcaga
     2101 cggctggaaa atctgatcgc ccagctgccc ggcgagaaga agaatggcct gttcggaaac
     2161 ctgattgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag
     2221 gatgccaaac tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc
     2281 cagatcggcg accagtacgc cgacctgttt ctggccgcca agaacctgtc cgacgccatc
     2341 ctgctgagcg acatcctgag agtgaacacc gagatcacca aggcccccct gagcgcctct
     2401 atgatcaaga gatacgacga gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg
     2461 cagcagctgc ctgagaagta caaagagatt ttcttcgacc agagcaagaa cggctacgcc
     2521 ggctacattg acggcggagc cagccaggaa gagttctaca agttcatcaa gcccatcctg
     2581 gaaaagatgg acggcaccga ggaactgctc gtgaagctga acagagagga cctgctgcgg
     2641 aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg agagctgcac
     2701 gccattctgc ggcggcagga agatttttac ccattcctga aggacaaccg ggaaaagatc
     2761 gagaagatcc tgaccttccg catcccctac tacgtgggcc ctctggccag gggaaacagc
     2821 agattcgcct ggatgaccag aaagagcgag gaaaccatca ccccctggaa cttcgaggaa
     2881 gtggtggaca agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa cttcgataag
     2941 aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg
     3001 tataacgagc tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc cgccttcctg
     3061 agcggcgagc agaaaaaggc catcgtggac ctgctgttca agaccaaccg gaaagtgacc
     3121 gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc
     3181 tccggcgtgg aagatcggtt caacgcctcc ctgggcacat accacgatct gctgaaaatt
     3241 atcaaggaca aggacttcct ggacaatgag gaaaacgagg acattctgga agatatcgtg
     3301 ctgaccctga cactgtttga ggacagagag atgatcgagg aacggctgaa aacctatgcc
     3361 cacctgttcg acgacaaagt gatgaagcag ctgaagcggc ggagatacac cggctggggc
     3421 aggctgagcc ggaagctgat caacggcatc cgggacaagc agtccggcaa gacaatcctg
     3481 gatttcctga agtccgacgg cttcgccaac agaaacttca tgcagctgat ccacgacgac
     3541 agcctgacct ttaaagagga catccagaaa gcccaggtgt ccggccaggg cgatagcctg
     3601 cacgagcaca ttgccaatct ggccggcagc cccgccatta agaagggcat cctgcagaca
     3661 gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc acaagcccga gaacatcgtg
     3721 atcgaaatgg ccagagagaa ccagaccacc cagaagggac agaagaacag ccgcgagaga
     3781 atgaagcgga tcgaagaggg catcaaagag ctgggcagcc agatcctgaa agaacacccc
     3841 gtggaaaaca cccagctgca gaacgagaag ctgtacctgt actacctgca gaatgggcgg
     3901 gatatgtacg tggaccagga actggacatc aaccggctgt ccgactacga tgtggaccat
     3961 atcgtgcctc agagctttct gaaggacgac tccatcgaca acaaggtgct gaccagaagc
     4021 gacaagaacc ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag
     4081 aactactggc ggcagctgct gaacgccaag ctgattaccc agagaaagtt cgacaatctg
     4141 accaaggccg agagaggcgg cctgagcgaa ctggataagg ccggcttcat caagagacag
     4201 ctggtggaaa cccggcagat cacaaagcac gtggcacaga tcctggactc ccggatgaac
     4261 actaagtacg acgagaatga caagctgatc cgggaagtga aagtgatcac cctgaagtcc
     4321 aagctggtgt ccgatttccg gaaggatttc cagttttaca aagtgcgcga gatcaacaac
     4381 taccaccacg cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag
     4441 taccctaagc tggaaagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag
     4501 atgatcgcca agagcgagca ggaaatcggc aaggctaccg ccaagtactt cttctacagc
     4561 aacatcatga actttttcaa gaccgagatt accctggcca acggcgagat ccggaagcgg
     4621 cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt gggataaggg ccgggatttt
     4681 gccaccgtgc ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg
     4741 cagacaggcg gcttcagcaa agagtctatc ctgcccaaga ggaacagcga taagctgatc
     4801 gccagaaaga aggactggga ccctaagaag tacggcggct tcgacagccc caccgtggcc
     4861 tattctgtgc tggtggtggc caaagtggaa aagggcaagt ccaagaaact gaagagtgtg
     4921 aaagagctgc tggggatcac catcatggaa agaagcagct tcgagaagaa tcccatcgac
     4981 tttctggaag ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag
     5041 tactccctgt tcgagctgga aaacggccgg aagagaatgc tggcctctgc cggcgaactg
     5101 cagaagggaa acgaactggc cctgccctcc aaatatgtga acttcctgta cctggccagc
     5161 cactatgaga agctgaaggg ctcccccgag gataatgagc agaaacagct gtttgtggaa
     5221 cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttctc caagagagtg
     5281 atcctggccg acgctaatct ggacaaagtg ctgtccgcct acaacaagca ccgggataag
     5341 cccatcagag agcaggccga gaatatcatc cacctgttta ccctgaccaa tctgggagcc
     5401 cctgccgcct tcaagtactt tgacaccacc atcgaccgga agaggtacac cagcaccaaa
     5461 gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga gacacggatc
     5521 gacctgtctc agctgggagg cgacagcccc aagaagaaga gaaaggtgga ggccagctaa
     5581 gaattctaac tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     5641 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     5701 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     5761 gtggggtggg gcaggacagc aagggggagg attgggaaga gaatagcagg catgctgggg
     5821 agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     5881 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     5941 tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc
     6001 atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg
     6061 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     6121 cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     6181 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     6241 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     6301 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     6361 tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     6421 ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     6481 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata
     6541 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     6601 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     6661 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     6721 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     6781 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     6841 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     6901 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     6961 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     7021 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     7081 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     7141 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     7201 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     7261 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     7321 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     7381 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     7441 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     7501 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     7561 tggctggttt attgctgata aatctggagc cggtgagcgt ggaagccgcg gtatcattgc
     7621 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     7681 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     7741 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     7801 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     7861 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     7921 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     7981 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     8041 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     8101 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     8161 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     8221 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     8281 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     8341 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     8401 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     8461 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     8521 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtgag ggcctatttc
     8581 ccatgattcc ttcatatttg catatacga
//
