LOCUS       sequence-291892-        9263 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Endogenous tagging of CaV-beta-3: C-terminal (amino acid position: 
            W479).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9263)
  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 9263)
  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 Cacnb3-GFP KI
FEATURES             Location/Qualifiers
     source          1..9263
                     /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        309..384
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             join(515..517,518..1228)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(515..517,518..1228)
                     /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(818..837)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1165..1186
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     enhancer        1305..1590
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        1592..1869
                     /label=chicken beta-actin promoter
     intron          1870..2097
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             2118..2144
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2148..2168
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2178..6278
                     /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             2178..6278
                     /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             6282..6302
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6342..6359)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6348..6555
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6564..6704
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6564..6693
                     /label=AAV2 ITR
     rep_origin      6779..7234
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6866..6885)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7076..7097
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7251..7270)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7370..7392
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7430..7448)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7516..7620
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7621..7689,7690..8481)
                     /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(7839..7858)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8652..9240
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9141..9160
                     /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 aaacacccca gtagctgtcc
      301 ttaggccagt tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt
      361 gaaaaagtgg caccgagtcg gtgctttttt gttttagagc tagaaatagc aagttaaaat
      421 aaggctagtc cgtttttagc gcgtgcgcca attctgcaga caaatggctc tagaagcttc
      481 agtagctgtc cttaggccag ggagtgctag cggagtgagc aagggcgagg agctgttcac
      541 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
      601 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
      661 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
      721 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
      781 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
      841 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
      901 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
      961 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     1021 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     1081 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     1141 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     1201 cactctcggc atggacgagc tgtacaagag cgcttgacag tagctgtcct taggccaggg
     1261 ctcgaggatc ctaggacgcg tcctaggacg cgtccagagg tacccgttac ataacttacg
     1321 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aatagtaacg
     1381 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     1441 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     1501 tggcccgcct ggcattgtgc ccagtacatg accttatggg actttcctac ttggcagtac
     1561 atctacgtat tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact
     1621 ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt
     1681 tgtgcagcga tgggggcggg gggggggggg gggcgcgcgc caggcggggc ggggcggggc
     1741 gaggggcggg gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc
     1801 cgaaagtttc cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg
     1861 cggcgggcgg gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg
     1921 cgccgcccgc cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc
     1981 ccttctcctc cgggctgtaa ttagctgagc aagaggtaag ggtttaaggg atggttggtt
     2041 ggtggggtat taatgtttaa ttacctggag cacctgcctg aaatcacttt ttttcaggtt
     2101 ggaccggtgc caccatgtac ccatacgatg ttccagatta cgcttcgccg aagaaaaagc
     2161 gcaaggtcga agcgtccgac aagaagtaca gcatcggcct ggacatcggc accaactctg
     2221 tgggctgggc cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg
     2281 gcaacaccga ccggcacagc atcaagaaga acctgatcgg agccctgctg ttcgacagcg
     2341 gcgaaacagc cgaggccacc cggctgaaga gaaccgccag aagaagatac accagacgga
     2401 agaaccggat ctgctatctg caagagatct tcagcaacga gatggccaag gtggacgaca
     2461 gcttcttcca cagactggaa gagtccttcc tggtggaaga ggataagaag cacgagcggc
     2521 accccatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac cccaccatct
     2581 accacctgag aaagaaactg gtggacagca ccgacaaggc cgacctgcgg ctgatctatc
     2641 tggccctggc ccacatgatc aagttccggg gccacttcct gatcgagggc gacctgaacc
     2701 ccgacaacag cgacgtggac aagctgttca tccagctggt gcagacctac aaccagctgt
     2761 tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa ggccatcctg tctgccagac
     2821 tgagcaagag cagacggctg gaaaatctga tcgcccagct gcccggcgag aagaagaatg
     2881 gcctgttcgg aaacctgatt gccctgagcc tgggcctgac ccccaacttc aagagcaact
     2941 tcgacctggc cgaggatgcc aaactgcagc tgagcaagga cacctacgac gacgacctgg
     3001 acaacctgct ggcccagatc ggcgaccagt acgccgacct gtttctggcc gccaagaacc
     3061 tgtccgacgc catcctgctg agcgacatcc tgagagtgaa caccgagatc accaaggccc
     3121 ccctgagcgc ctctatgatc aagagatacg acgagcacca ccaggacctg accctgctga
     3181 aagctctcgt gcggcagcag ctgcctgaga agtacaaaga gattttcttc gaccagagca
     3241 agaacggcta cgccggctac attgacggcg gagccagcca ggaagagttc tacaagttca
     3301 tcaagcccat cctggaaaag atggacggca ccgaggaact gctcgtgaag ctgaacagag
     3361 aggacctgct gcggaagcag cggaccttcg acaacggcag catcccccac cagatccacc
     3421 tgggagagct gcacgccatt ctgcggcggc aggaagattt ttacccattc ctgaaggaca
     3481 accgggaaaa gatcgagaag atcctgacct tccgcatccc ctactacgtg ggccctctgg
     3541 ccaggggaaa cagcagattc gcctggatga ccagaaagag cgaggaaacc atcaccccct
     3601 ggaacttcga ggaagtggtg gacaagggcg cttccgccca gagcttcatc gagcggatga
     3661 ccaacttcga taagaacctg cccaacgaga aggtgctgcc caagcacagc ctgctgtacg
     3721 agtacttcac cgtgtataac gagctgacca aagtgaaata cgtgaccgag ggaatgagaa
     3781 agcccgcctt cctgagcggc gagcagaaaa aggccatcgt ggacctgctg ttcaagacca
     3841 accggaaagt gaccgtgaag cagctgaaag aggactactt caagaaaatc gagtgcttcg
     3901 actccgtgga aatctccggc gtggaagatc ggttcaacgc ctccctgggc acataccacg
     3961 atctgctgaa aattatcaag gacaaggact tcctggacaa tgaggaaaac gaggacattc
     4021 tggaagatat cgtgctgacc ctgacactgt ttgaggacag agagatgatc gaggaacggc
     4081 tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag cggcggagat
     4141 acaccggctg gggcaggctg agccggaagc tgatcaacgg catccgggac aagcagtccg
     4201 gcaagacaat cctggatttc ctgaagtccg acggcttcgc caacagaaac ttcatgcagc
     4261 tgatccacga cgacagcctg acctttaaag aggacatcca gaaagcccag gtgtccggcc
     4321 agggcgatag cctgcacgag cacattgcca atctggccgg cagccccgcc attaagaagg
     4381 gcatcctgca gacagtgaag gtggtggacg agctcgtgaa agtgatgggc cggcacaagc
     4441 ccgagaacat cgtgatcgaa atggccagag agaaccagac cacccagaag ggacagaaga
     4501 acagccgcga gagaatgaag cggatcgaag agggcatcaa agagctgggc agccagatcc
     4561 tgaaagaaca ccccgtggaa aacacccagc tgcagaacga gaagctgtac ctgtactacc
     4621 tgcagaatgg gcgggatatg tacgtggacc aggaactgga catcaaccgg ctgtccgact
     4681 acgatgtgga ccatatcgtg cctcagagct ttctgaagga cgactccatc gacaacaagg
     4741 tgctgaccag aagcgacaag aaccggggca agagcgacaa cgtgccctcc gaagaggtcg
     4801 tgaagaagat gaagaactac tggcggcagc tgctgaacgc caagctgatt acccagagaa
     4861 agttcgacaa tctgaccaag gccgagagag gcggcctgag cgaactggat aaggccggct
     4921 tcatcaagag acagctggtg gaaacccggc agatcacaaa gcacgtggca cagatcctgg
     4981 actcccggat gaacactaag tacgacgaga atgacaagct gatccgggaa gtgaaagtga
     5041 tcaccctgaa gtccaagctg gtgtccgatt tccggaagga tttccagttt tacaaagtgc
     5101 gcgagatcaa caactaccac cacgcccacg acgcctacct gaacgccgtc gtgggaaccg
     5161 ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt
     5221 acgacgtgcg gaagatgatc gccaagagcg agcaggaaat cggcaaggct accgccaagt
     5281 acttcttcta cagcaacatc atgaactttt tcaagaccga gattaccctg gccaacggcg
     5341 agatccggaa gcggcctctg atcgagacaa acggcgaaac cggggagatc gtgtgggata
     5401 agggccggga ttttgccacc gtgcggaaag tgctgagcat gccccaagtg aatatcgtga
     5461 aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc aagaggaaca
     5521 gcgataagct gatcgccaga aagaaggact gggaccctaa gaagtacggc ggcttcgaca
     5581 gccccaccgt ggcctattct gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga
     5641 aactgaagag tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc agcttcgaga
     5701 agaatcccat cgactttctg gaagccaagg gctacaaaga agtgaaaaag gacctgatca
     5761 tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg ccggaagaga atgctggcct
     5821 ctgccggcga actgcagaag ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc
     5881 tgtacctggc cagccactat gagaagctga agggctcccc cgaggataat gagcagaaac
     5941 agctgtttgt ggaacagcac aagcactacc tggacgagat catcgagcag atcagcgagt
     6001 tctccaagag agtgatcctg gccgacgcta atctggacaa agtgctgtcc gcctacaaca
     6061 agcaccggga taagcccatc agagagcagg ccgagaatat catccacctg tttaccctga
     6121 ccaatctggg agcccctgcc gccttcaagt actttgacac caccatcgac cggaagaggt
     6181 acaccagcac caaagaggtg ctggacgcca ccctgatcca ccagagcatc accggcctgt
     6241 acgagacacg gatcgacctg tctcagctgg gaggcgacag ccccaagaag aagagaaagg
     6301 tggaggccag ctaagaattc taactagagc tcgctgatca gcctcgactg tgccttctag
     6361 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     6421 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     6481 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagagaatag
     6541 caggcatgct ggggagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct
     6601 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc
     6661 ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta
     6721 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac
     6781 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     6841 acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     6901 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     6961 gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca
     7021 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     7081 ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa
     7141 gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     7201 gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc
     7261 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     7321 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     7381 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     7441 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     7501 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     7561 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     7621 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     7681 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     7741 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     7801 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     7861 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     7921 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     7981 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     8041 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     8101 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     8161 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     8221 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     8281 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtggaagc
     8341 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     8401 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     8461 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     8521 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     8581 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     8641 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     8701 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     8761 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     8821 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     8881 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     8941 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     9001 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     9061 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     9121 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     9181 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     9241 aacgccagca acgcggcctt ttt
//
