LOCUS       sequence-252018-        9253 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Endogenous tagging of beta-actin: N-terminal (amino acid position: 
            D2).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9253)
  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 9253)
  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
FEATURES             Location/Qualifiers
     source          1..9253
                     /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        307..382
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             join(506..508,509..1219)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(506..508,509..1219)
                     /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(809..828)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1156..1177
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     enhancer        1295..1580
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        1582..1859
                     /label=chicken beta-actin promoter
     intron          1860..2087
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             2108..2134
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2138..2158
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2168..6268
                     /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             2168..6268
                     /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             6272..6292
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6332..6349)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6338..6545
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6554..6694
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6554..6683
                     /label=AAV2 ITR
     rep_origin      6769..7224
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6856..6875)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7066..7087
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7241..7260)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7360..7382
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7420..7438)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7506..7610
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7611..7679,7680..8471)
                     /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(7829..7848)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8642..9230
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9131..9150
                     /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 aaacaccgca gcgatatcgt
      301 catccagttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga
      361 aaaagtggca ccgagtcggt gcttttttgt tttagagcta gaaatagcaa gttaaaataa
      421 ggctagtccg tttttagcgc gtgcgccaat tctgcagaca aatggctcta gaagcttcgc
      481 agcgatatcg tcatccaggg ctagcgtgag caagggcgag gagctgttca ccggggtggt
      541 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
      601 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
      661 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
      721 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
      781 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
      841 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
      901 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
      961 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     1021 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     1081 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     1141 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     1201 catggacgag ctgtacaagt caggcagcgc agcgatatcg tcatccaggg ctcgaggatc
     1261 ctaggacgcg tcctaggacg cgtccagagg tacccgttac ataacttacg gtaaatggcc
     1321 cgcctggctg accgcccaac gacccccgcc cattgacgtc aatagtaacg ccaataggga
     1381 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     1441 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     1501 ggcattgtgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     1561 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
     1621 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
     1681 tgggggcggg gggggggggg gggcgcgcgc caggcggggc ggggcggggc gaggggcggg
     1741 gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc
     1801 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
     1861 gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc
     1921 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
     1981 cgggctgtaa ttagctgagc aagaggtaag ggtttaaggg atggttggtt ggtggggtat
     2041 taatgtttaa ttacctggag cacctgcctg aaatcacttt ttttcaggtt ggaccggtgc
     2101 caccatgtac ccatacgatg ttccagatta cgcttcgccg aagaaaaagc gcaaggtcga
     2161 agcgtccgac aagaagtaca gcatcggcct ggacatcggc accaactctg tgggctgggc
     2221 cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga
     2281 ccggcacagc atcaagaaga acctgatcgg agccctgctg ttcgacagcg gcgaaacagc
     2341 cgaggccacc cggctgaaga gaaccgccag aagaagatac accagacgga agaaccggat
     2401 ctgctatctg caagagatct tcagcaacga gatggccaag gtggacgaca gcttcttcca
     2461 cagactggaa gagtccttcc tggtggaaga ggataagaag cacgagcggc accccatctt
     2521 cggcaacatc gtggacgagg tggcctacca cgagaagtac cccaccatct accacctgag
     2581 aaagaaactg gtggacagca ccgacaaggc cgacctgcgg ctgatctatc tggccctggc
     2641 ccacatgatc aagttccggg gccacttcct gatcgagggc gacctgaacc ccgacaacag
     2701 cgacgtggac aagctgttca tccagctggt gcagacctac aaccagctgt tcgaggaaaa
     2761 ccccatcaac gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag
     2821 cagacggctg gaaaatctga tcgcccagct gcccggcgag aagaagaatg gcctgttcgg
     2881 caacctgatt gccctgagcc tgggcctgac ccccaacttc aagagcaact tcgacctggc
     2941 cgaggatgcc aaactgcagc tgagcaagga cacctacgac gacgacctgg acaacctgct
     3001 ggcccagatc ggcgaccagt acgccgacct gtttctggcc gccaagaacc tgtccgacgc
     3061 catcctgctg agcgacatcc tgagagtgaa caccgagatc accaaggccc ccctgagcgc
     3121 ctctatgatc aagagatacg acgagcacca ccaggacctg accctgctga aagctctcgt
     3181 gcggcagcag ctgcctgaga agtacaaaga gattttcttc gaccagagca agaacggcta
     3241 cgccggctac attgacggcg gagccagcca ggaagagttc tacaagttca tcaagcccat
     3301 cctggaaaag atggacggca ccgaggaact gctcgtgaag ctgaacagag aggacctgct
     3361 gcggaagcag cggaccttcg acaacggcag catcccccac cagatccacc tgggagagct
     3421 gcacgccatt ctgcggcggc aggaagattt ttacccattc ctgaaggaca accgggaaaa
     3481 gatcgagaag atcctgacct tccgcatccc ctactacgtg ggccctctgg ccaggggaaa
     3541 cagcagattc gcctggatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga
     3601 ggaagtggtg gacaagggcg cttccgccca gagcttcatc gagcggatga ccaacttcga
     3661 taagaacctg cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac
     3721 cgtgtataac gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt
     3781 cctgagcggc gagcagaaaa aggccatcgt ggacctgctg ttcaagacca accggaaagt
     3841 gaccgtgaag cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga
     3901 aatctccggc gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa
     3961 aattatcaag gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat
     4021 cgtgctgacc ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta
     4081 tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag cggcggagat acaccggctg
     4141 gggcaggctg agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat
     4201 cctggatttc ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga
     4261 cgacagcctg acctttaaag aggacatcca gaaagcccag gtgtccggcc agggcgatag
     4321 cctgcacgag cacattgcca atctggccgg cagccccgcc attaagaagg gcatcctgca
     4381 gacagtgaag gtggtggacg agctcgtgaa agtgatgggc cggcacaagc ccgagaacat
     4441 cgtgatcgaa atggccagag agaaccagac cacccagaag ggacagaaga acagccgcga
     4501 gagaatgaag cggatcgaag agggcatcaa agagctgggc agccagatcc tgaaagaaca
     4561 ccccgtggaa aacacccagc tgcagaacga gaagctgtac ctgtactacc tgcagaatgg
     4621 gcgggatatg tacgtggacc aggaactgga catcaaccgg ctgtccgact acgatgtgga
     4681 ccatatcgtg cctcagagct ttctgaagga cgactccatc gacaacaagg tgctgaccag
     4741 aagcgacaag aaccggggca agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat
     4801 gaagaactac tggcggcagc tgctgaacgc caagctgatt acccagagaa agttcgacaa
     4861 tctgaccaag gccgagagag gcggcctgag cgaactggat aaggccggct tcatcaagag
     4921 acagctggtg gaaacccggc agatcacaaa gcacgtggca cagatcctgg actcccggat
     4981 gaacactaag tacgacgaga atgacaagct gatccgggaa gtgaaagtga tcaccctgaa
     5041 gtccaagctg gtgtccgatt tccggaagga tttccagttt tacaaagtgc gcgagatcaa
     5101 caactaccac cacgcccacg acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa
     5161 aaagtaccct aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg
     5221 gaagatgatc gccaagagcg agcaggaaat cggcaaggct accgccaagt acttcttcta
     5281 cagcaacatc atgaactttt tcaagaccga gattaccctg gccaacggcg agatccggaa
     5341 gcggcctctg atcgagacaa acggcgaaac cggggagatc gtgtgggata agggccggga
     5401 ttttgccacc gtgcggaaag tgctgagcat gccccaagtg aatatcgtga aaaagaccga
     5461 ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc aagaggaaca gcgataagct
     5521 gatcgccaga aagaaggact gggaccctaa gaagtacggc ggcttcgaca gccccaccgt
     5581 ggcctattct gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga aactgaagag
     5641 tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc agcttcgaga agaatcccat
     5701 cgactttctg gaagccaagg gctacaaaga agtgaaaaag gacctgatca tcaagctgcc
     5761 taagtactcc ctgttcgagc tggaaaacgg ccggaagaga atgctggcct ctgccggcga
     5821 actgcagaag ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc tgtacctggc
     5881 cagccactat gagaagctga agggctcccc cgaggataat gagcagaaac agctgtttgt
     5941 ggaacagcac aagcactacc tggacgagat catcgagcag atcagcgagt tctccaagag
     6001 agtgatcctg gccgacgcta atctggacaa agtgctgtcc gcctacaaca agcaccggga
     6061 taagcccatc agagagcagg ccgagaatat catccacctg tttaccctga ccaatctggg
     6121 agcccctgcc gccttcaagt actttgacac caccatcgac cggaagaggt acaccagcac
     6181 caaagaggtg ctggacgcca ccctgatcca ccagagcatc accggcctgt acgagacacg
     6241 gatcgacctg tctcagctgg gaggcgacag ccccaagaag aagagaaagg tggaggccag
     6301 ctaagaattc taactagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca
     6361 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     6421 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     6481 gggggtgggg tggggcagga cagcaagggg gaggattggg aagagaatag caggcatgct
     6541 ggggagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     6601 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     6661 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     6721 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     6781 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     6841 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     6901 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     6961 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     7021 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     7081 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
     7141 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     7201 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     7261 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     7321 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     7381 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     7441 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     7501 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     7561 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     7621 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     7681 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     7741 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     7801 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     7861 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     7921 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     7981 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     8041 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     8101 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     8161 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     8221 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     8281 cggctggctg gtttattgct gataaatctg gagccggtga gcgtggaagc cgcggtatca
     8341 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     8401 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     8461 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     8521 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     8581 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     8641 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     8701 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     8761 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     8821 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     8881 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     8941 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     9001 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     9061 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     9121 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     9181 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     9241 acgcggcctt ttt
//
