LOCUS       sequence-252056-        9270 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Endogenous tagging of WASP1/WAVE1: C-terminal (amino acid position: 
            STOP codon).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9270)
  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 9270)
  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 WASF1-GFP KI
FEATURES             Location/Qualifiers
     source          1..9270
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        62..310
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     62..82
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     233..252
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        331..406
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             join(551..553,554..1267)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(551..553,554..1264)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(854..873)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1201..1222
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     enhancer        1335..1620
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        1622..1899
                     /label=chicken beta-actin promoter
     intron          1900..2127
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             2148..2174
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2178..2198
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2208..6308
                     /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             2208..6308
                     /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             6312..6332
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6372..6389)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6378..6585
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6594..6734
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6594..6723
                     /label=AAV2 ITR
     rep_origin      6809..7264
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6896..6915)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7106..7127
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7281..7300)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7400..7422
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7460..7478)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7546..7650
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7651..7719,7720..8511)
                     /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(7869..7888)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8682..9270
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9171..9190
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 tgagggccta tttcccatga ttccttcata tttgcatata cgatacaagg ctgttagaga
      121 gataattgga attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag
      181 aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca
      241 tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg
      301 acgaaacacc gtagactggc tggagtgaga gttttagagc tagaaatagc aagttaaaat
      361 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgttttaga
      421 gctagaaata gcaagttaaa ataaggctag tccgttttta gcgcgtgcgc caattctgca
      481 gacaaatggc tctagaagct tccctctcac tccagccagt ctacaaggta gcggctccag
      541 tagatctggg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
      601 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
      661 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
      721 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
      781 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
      841 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
      901 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
      961 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     1021 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     1081 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     1141 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     1201 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     1261 caagtaaccc tctcactcca gccagtctac ctcgaggatc ctaggacgcg tcctaggacg
     1321 cgtccagagg tacccgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     1381 gacccccgcc cattgacgtc aatagtaacg ccaataggga ctttccattg acgtcaatgg
     1441 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     1501 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattgtgc ccagtacatg
     1561 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     1621 gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc cccaccccca
     1681 attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg gggggggggg
     1741 gggcgcgcgc caggcggggc ggggcggggc gaggggcggg gcggggcgag gcggagaggt
     1801 gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg
     1861 cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt
     1921 cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc cccggctctg actgaccgcg
     1981 ttactcccac aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa ttagctgagc
     2041 aagaggtaag ggtttaaggg atggttggtt ggtggggtat taatgtttaa ttacctggag
     2101 cacctgcctg aaatcacttt ttttcaggtt ggaccggtgc caccatgtac ccatacgatg
     2161 ttccagatta cgcttcgccg aagaaaaagc gcaaggtcga agcgtccgac aagaagtaca
     2221 gcatcggcct ggacatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca
     2281 aggtgcccag caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga
     2341 acctgatcgg agccctgctg ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga
     2401 gaaccgccag aagaagatac accagacgga agaaccggat ctgctatctg caagagatct
     2461 tcagcaacga gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc
     2521 tggtggaaga ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg
     2581 tggcctacca cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca
     2641 ccgacaaggc cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg
     2701 gccacttcct gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca
     2761 tccagctggt gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg
     2821 tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga
     2881 tcgcccagct gcccggcgag aagaagaatg gcctgttcgg caacctgatt gccctgagcc
     2941 tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc
     3001 tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt
     3061 acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc
     3121 tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg
     3181 acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga
     3241 agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac attgacggcg
     3301 gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca
     3361 ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg
     3421 acaacggcag catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc
     3481 aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct
     3541 tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga
     3601 ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg
     3661 cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga
     3721 aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac gagctgacca
     3781 aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa
     3841 aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag
     3901 aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc
     3961 ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact
     4021 tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt
     4081 ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca
     4141 aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc
     4201 tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg
     4261 acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag
     4321 aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca
     4381 atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg
     4441 agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag
     4501 agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag
     4561 agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc
     4621 tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc
     4681 aggaactgga catcaaccgg ctgtccgact acgatgtgga ccatatcgtg cctcagagct
     4741 ttctgaagga cgactccatc gacaacaagg tgctgaccag aagcgacaag aaccggggca
     4801 agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcggcagc
     4861 tgctgaacgc caagctgatt acccagagaa agttcgacaa tctgaccaag gccgagagag
     4921 gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc
     4981 agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga
     5041 atgacaagct gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt
     5101 tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg
     5161 acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa
     5221 gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg
     5281 agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt
     5341 tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa
     5401 acggcgaaac cggggagatc gtgtgggata agggccggga ttttgccacc gtgcggaaag
     5461 tgctgagcat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca
     5521 gcaaagagtc tatcctgccc aagaggaaca gcgataagct gatcgccaga aagaaggact
     5581 gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct gtgctggtgg
     5641 tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctgctgggga
     5701 tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg
     5761 gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc
     5821 tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag ggaaacgaac
     5881 tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga
     5941 agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac aagcactacc
     6001 tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta
     6061 atctggacaa agtgctgtcc gcctacaaca agcaccggga taagcccatc agagagcagg
     6121 ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt
     6181 actttgacac caccatcgac cggaagaggt acaccagcac caaagaggtg ctggacgcca
     6241 ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg
     6301 gaggcgacag ccccaagaag aagagaaagg tggaggccag ctaagaattc taactagagc
     6361 tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     6421 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     6481 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     6541 cagcaagggg gaggattggg aagagaatag caggcatgct ggggagcggc cgcaggaacc
     6601 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     6661 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     6721 cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     6781 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
     6841 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct
     6901 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     6961 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     7021 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     7081 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     7141 tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta
     7201 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     7261 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     7321 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     7381 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     7441 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     7501 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     7561 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     7621 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     7681 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     7741 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     7801 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     7861 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     7921 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     7981 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     8041 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     8101 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     8161 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     8221 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     8281 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     8341 gataaatctg gagccggtga gcgtggaagc cgcggtatca ttgcagcact ggggccagat
     8401 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     8461 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     8521 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     8581 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     8641 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     8701 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     8761 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     8821 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     8881 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     8941 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     9001 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     9061 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     9121 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     9181 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     9241 tgctcgtcag gggggcggag cctatggaaa
//
