LOCUS       sequence-291869-        9299 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Endogenous tagging of beta-actin: N-terminal (amino acid position: 
            before startcodon).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9299)
  AUTHORS   Willems J, de Jong APH, Scheefhals N, Mertens E, Catsburg LAE, 
            Poorthuis RB, de Winter F, Verhaagen J, Meye FJ, MacGillavry HD
  TITLE     ORANGE: A CRISPR/Cas9-based genome editing toolbox for epitope 
            tagging of endogenous proteins in neurons.
  JOURNAL   PLoS Biol. 2020 Apr 10;18(4):e3000665. doi: 
            10.1371/journal.pbio.3000665.
  PUBMED    32275651
REFERENCE   2  (bases 1 to 9299)
  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: "PLoS 
            Biol."; date: "2020-04-10"; volume: "18(4):e3000665. doi"; pages: " 
            10.1371/journal.pbio.3000665"
COMMENT     Sequence Label: pORANGE GFP-Actb KI #2
FEATURES             Location/Qualifiers
     source          1..9299
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        39..287
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     39..59
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     210..229
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        308..383
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             join(534..536,537..1247)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(534..536,537..1247)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(837..856)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1184..1205
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     enhancer        1341..1626
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        1628..1905
                     /label=chicken beta-actin promoter
     intron          1906..2133
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             2154..2180
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2184..2204
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2214..6314
                     /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             2214..6314
                     /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             6318..6338
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6378..6395)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6384..6591
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6600..6740
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6600..6729
                     /label=AAV2 ITR
     rep_origin      6815..7270
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6902..6921)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7112..7133
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7287..7306)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7406..7428
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7466..7484)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7552..7656
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7657..7725,7726..8517)
                     /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(7875..7894)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8688..9276
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9177..9196
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acggttcctg gccttttgct ggccttttgc tcacatgtga gggcctattt cccatgattc
       61 cttcatattt gcatatacga tacaaggctg ttagagagat aattggaatt aatttgactg
      121 taaacacaaa gatattagta caaaatacgt gacgtagaaa gtaataattt cttgggtagt
      181 ttgcagtttt aaaattatgt tttaaaatgg actatcatat gcttaccgta acttgaaagt
      241 atttcgattt cttggcttta tatatcttgt ggaaaggacg aaacacctgt gccttgatag
      301 ttcgccagtt ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg
      361 aaaaagtggc accgagtcgg tgcttttttg ttttagagct agaaatagca agttaaaata
      421 aggctagtcc gtttttagcg cgtgcgccaa ttctgcagac aaatggctct agaagctttc
      481 gaagacccta gaccctggcg aactatcaag gcacattcgc catggctagc ggagtgagca
      541 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
      601 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
      661 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
      721 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
      781 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
      841 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
      901 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
      961 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     1021 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     1081 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     1141 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     1201 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagagt ggaagcgctc
     1261 cctggcgaac tatcaaggca cactcggggt cttctcctcg aggatcctag gacgcgtcct
     1321 aggacgcgtc cagaggtacc cgttacataa cttacggtaa atggcccgcc tggctgaccg
     1381 cccaacgacc cccgcccatt gacgtcaata gtaacgccaa tagggacttt ccattgacgt
     1441 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     1501 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttgtgcccag
     1561 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     1621 accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca
     1681 cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg
     1741 gggggggggc gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg ggcgaggcgg
     1801 agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg
     1861 cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt cgctgcgcgc
     1921 tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg
     1981 accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag
     2041 ctgagcaaga ggtaagggtt taagggatgg ttggttggtg gggtattaat gtttaattac
     2101 ctggagcacc tgcctgaaat cacttttttt caggttggac cggtgccacc atgtacccat
     2161 acgatgttcc agattacgct tcgccgaaga aaaagcgcaa ggtcgaagcg tccgacaaga
     2221 agtacagcat cggcctggac atcggcacca actctgtggg ctgggccgtg atcaccgacg
     2281 agtacaaggt gcccagcaag aaattcaagg tgctgggcaa caccgaccgg cacagcatca
     2341 agaagaacct gatcggagcc ctgctgttcg acagcggcga aacagccgag gccacccggc
     2401 tgaagagaac cgccagaaga agatacacca gacggaagaa ccggatctgc tatctgcaag
     2461 agatcttcag caacgagatg gccaaggtgg acgacagctt cttccacaga ctggaagagt
     2521 ccttcctggt ggaagaggat aagaagcacg agcggcaccc catcttcggc aacatcgtgg
     2581 acgaggtggc ctaccacgag aagtacccca ccatctacca cctgagaaag aaactggtgg
     2641 acagcaccga caaggccgac ctgcggctga tctatctggc cctggcccac atgatcaagt
     2701 tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac gtggacaagc
     2761 tgttcatcca gctggtgcag acctacaacc agctgttcga ggaaaacccc atcaacgcca
     2821 gcggcgtgga cgccaaggcc atcctgtctg ccagactgag caagagcaga cggctggaaa
     2881 atctgatcgc ccagctgccc ggcgagaaga agaatggcct gttcggaaac ctgattgccc
     2941 tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag gatgccaaac
     3001 tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc cagatcggcg
     3061 accagtacgc cgacctgttt ctggccgcca agaacctgtc cgacgccatc ctgctgagcg
     3121 acatcctgag agtgaacacc gagatcacca aggcccccct gagcgcctct atgatcaaga
     3181 gatacgacga gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc
     3241 ctgagaagta caaagagatt ttcttcgacc agagcaagaa cggctacgcc ggctacattg
     3301 acggcggagc cagccaggaa gagttctaca agttcatcaa gcccatcctg gaaaagatgg
     3361 acggcaccga ggaactgctc gtgaagctga acagagagga cctgctgcgg aagcagcgga
     3421 ccttcgacaa cggcagcatc ccccaccaga tccacctggg agagctgcac gccattctgc
     3481 ggcggcagga agatttttac ccattcctga aggacaaccg ggaaaagatc gagaagatcc
     3541 tgaccttccg catcccctac tacgtgggcc ctctggccag gggaaacagc agattcgcct
     3601 ggatgaccag aaagagcgag gaaaccatca ccccctggaa cttcgaggaa gtggtggaca
     3661 agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa cttcgataag aacctgccca
     3721 acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg tataacgagc
     3781 tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc cgccttcctg agcggcgagc
     3841 agaaaaaggc catcgtggac ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc
     3901 tgaaagagga ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg
     3961 aagatcggtt caacgcctcc ctgggcacat accacgatct gctgaaaatt atcaaggaca
     4021 aggacttcct ggacaatgag gaaaacgagg acattctgga agatatcgtg ctgaccctga
     4081 cactgtttga ggacagagag atgatcgagg aacggctgaa aacctatgcc cacctgttcg
     4141 acgacaaagt gatgaagcag ctgaagcggc ggagatacac cggctggggc aggctgagcc
     4201 ggaagctgat caacggcatc cgggacaagc agtccggcaa gacaatcctg gatttcctga
     4261 agtccgacgg cttcgccaac agaaacttca tgcagctgat ccacgacgac agcctgacct
     4321 ttaaagagga catccagaaa gcccaggtgt ccggccaggg cgatagcctg cacgagcaca
     4381 ttgccaatct ggccggcagc cccgccatta agaagggcat cctgcagaca gtgaaggtgg
     4441 tggacgagct cgtgaaagtg atgggccggc acaagcccga gaacatcgtg atcgaaatgg
     4501 ccagagagaa ccagaccacc cagaagggac agaagaacag ccgcgagaga atgaagcgga
     4561 tcgaagaggg catcaaagag ctgggcagcc agatcctgaa agaacacccc gtggaaaaca
     4621 cccagctgca gaacgagaag ctgtacctgt actacctgca gaatgggcgg gatatgtacg
     4681 tggaccagga actggacatc aaccggctgt ccgactacga tgtggaccat atcgtgcctc
     4741 agagctttct gaaggacgac tccatcgaca acaaggtgct gaccagaagc gacaagaacc
     4801 ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag aactactggc
     4861 ggcagctgct gaacgccaag ctgattaccc agagaaagtt cgacaatctg accaaggccg
     4921 agagaggcgg cctgagcgaa ctggataagg ccggcttcat caagagacag ctggtggaaa
     4981 cccggcagat cacaaagcac gtggcacaga tcctggactc ccggatgaac actaagtacg
     5041 acgagaatga caagctgatc cgggaagtga aagtgatcac cctgaagtcc aagctggtgt
     5101 ccgatttccg gaaggatttc cagttttaca aagtgcgcga gatcaacaac taccaccacg
     5161 cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc
     5221 tggaaagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag atgatcgcca
     5281 agagcgagca ggaaatcggc aaggctaccg ccaagtactt cttctacagc aacatcatga
     5341 actttttcaa gaccgagatt accctggcca acggcgagat ccggaagcgg cctctgatcg
     5401 agacaaacgg cgaaaccggg gagatcgtgt gggataaggg ccgggatttt gccaccgtgc
     5461 ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg
     5521 gcttcagcaa agagtctatc ctgcccaaga ggaacagcga taagctgatc gccagaaaga
     5581 aggactggga ccctaagaag tacggcggct tcgacagccc caccgtggcc tattctgtgc
     5641 tggtggtggc caaagtggaa aagggcaagt ccaagaaact gaagagtgtg aaagagctgc
     5701 tggggatcac catcatggaa agaagcagct tcgagaagaa tcccatcgac tttctggaag
     5761 ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag tactccctgt
     5821 tcgagctgga aaacggccgg aagagaatgc tggcctctgc cggcgaactg cagaagggaa
     5881 acgaactggc cctgccctcc aaatatgtga acttcctgta cctggccagc cactatgaga
     5941 agctgaaggg ctcccccgag gataatgagc agaaacagct gtttgtggaa cagcacaagc
     6001 actacctgga cgagatcatc gagcagatca gcgagttctc caagagagtg atcctggccg
     6061 acgctaatct ggacaaagtg ctgtccgcct acaacaagca ccgggataag cccatcagag
     6121 agcaggccga gaatatcatc cacctgttta ccctgaccaa tctgggagcc cctgccgcct
     6181 tcaagtactt tgacaccacc atcgaccgga agaggtacac cagcaccaaa gaggtgctgg
     6241 acgccaccct gatccaccag agcatcaccg gcctgtacga gacacggatc gacctgtctc
     6301 agctgggagg cgacagcccc aagaagaaga gaaaggtgga ggccagctaa gaattctaac
     6361 tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     6421 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     6481 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     6541 gcaggacagc aagggggagg attgggaaga gaatagcagg catgctgggg agcggccgca
     6601 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     6661 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     6721 agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     6781 tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc
     6841 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc
     6901 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     6961 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     7021 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     7081 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     7141 ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat
     7201 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     7261 tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     7321 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     7381 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     7441 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     7501 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     7561 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     7621 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     7681 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     7741 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     7801 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     7861 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     7921 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     7981 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     8041 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     8101 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     8161 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     8221 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     8281 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     8341 attgctgata aatctggagc cggtgagcgt ggaagccgcg gtatcattgc agcactgggg
     8401 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     8461 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     8521 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     8581 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     8641 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     8701 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     8761 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     8821 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     8881 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     8941 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     9001 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     9061 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     9121 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     9181 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     9241 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttt
//
