LOCUS       sequence-252020-        9252 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Endogenous tagging of CaV2.1, P/Q: N-terminal (amino acid position: 
            G10).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9252)
  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 9252)
  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-CACNA1A KI
FEATURES             Location/Qualifiers
     source          1..9252
                     /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(503..505,506..1216)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(503..505,506..1216)
                     /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(806..825)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1153..1174
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     enhancer        1294..1579
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        1581..1858
                     /label=chicken beta-actin promoter
     intron          1859..2086
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             2107..2133
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2137..2157
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2167..6267
                     /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             2167..6267
                     /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             6271..6291
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6331..6348)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6337..6544
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6553..6693
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6553..6682
                     /label=AAV2 ITR
     rep_origin      6768..7223
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6855..6874)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7065..7086
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7240..7259)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7359..7381
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7419..7437)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7505..7609
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7610..7678,7679..8470)
                     /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(7828..7847)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8641..9229
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9130..9149
                     /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 aaacaccgcc gcctgcgccg
      301 tagcggccgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt
      361 gaaaaagtgg caccgagtcg gtgctttttt gttttagagc tagaaatagc aagttaaaat
      421 aaggctagtc cgtttttagc gcgtgcgcca attctgcaga caaatggctc tagaagcttc
      481 cgcctgcgcc gtagcggccg gggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
      541 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
      601 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
      661 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
      721 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
      781 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
      841 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
      901 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
      961 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     1021 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     1081 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     1141 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     1201 ggacgagctg tacaagggtt caggggccgc ctgcgccgta gcggccgggc tcgaggatcc
     1261 taggacgcgt cctaggacgc gtccagaggt acccgttaca taacttacgg taaatggccc
     1321 gcctggctga ccgcccaacg acccccgccc attgacgtca atagtaacgc caatagggac
     1381 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     1441 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     1501 gcattgtgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     1561 agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc
     1621 ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat
     1681 gggggcgggg gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg
     1741 cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc
     1801 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg
     1861 agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc
     1921 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
     1981 gggctgtaat tagctgagca agaggtaagg gtttaaggga tggttggttg gtggggtatt
     2041 aatgtttaat tacctggagc acctgcctga aatcactttt tttcaggttg gaccggtgcc
     2101 accatgtacc catacgatgt tccagattac gcttcgccga agaaaaagcg caaggtcgaa
     2161 gcgtccgaca agaagtacag catcggcctg gacatcggca ccaactctgt gggctgggcc
     2221 gtgatcaccg acgagtacaa ggtgcccagc aagaaattca aggtgctggg caacaccgac
     2281 cggcacagca tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc
     2341 gaggccaccc ggctgaagag aaccgccaga agaagataca ccagacggaa gaaccggatc
     2401 tgctatctgc aagagatctt cagcaacgag atggccaagg tggacgacag cttcttccac
     2461 agactggaag agtccttcct ggtggaagag gataagaagc acgagcggca ccccatcttc
     2521 ggcaacatcg tggacgaggt ggcctaccac gagaagtacc ccaccatcta ccacctgaga
     2581 aagaaactgg tggacagcac cgacaaggcc gacctgcggc tgatctatct ggccctggcc
     2641 cacatgatca agttccgggg ccacttcctg atcgagggcg acctgaaccc cgacaacagc
     2701 gacgtggaca agctgttcat ccagctggtg cagacctaca accagctgtt cgaggaaaac
     2761 cccatcaacg ccagcggcgt ggacgccaag gccatcctgt ctgccagact gagcaagagc
     2821 agacggctgg aaaatctgat cgcccagctg cccggcgaga agaagaatgg cctgttcggc
     2881 aacctgattg ccctgagcct gggcctgacc cccaacttca agagcaactt cgacctggcc
     2941 gaggatgcca aactgcagct gagcaaggac acctacgacg acgacctgga caacctgctg
     3001 gcccagatcg gcgaccagta cgccgacctg tttctggccg ccaagaacct gtccgacgcc
     3061 atcctgctga gcgacatcct gagagtgaac accgagatca ccaaggcccc cctgagcgcc
     3121 tctatgatca agagatacga cgagcaccac caggacctga ccctgctgaa agctctcgtg
     3181 cggcagcagc tgcctgagaa gtacaaagag attttcttcg accagagcaa gaacggctac
     3241 gccggctaca ttgacggcgg agccagccag gaagagttct acaagttcat caagcccatc
     3301 ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg
     3361 cggaagcagc ggaccttcga caacggcagc atcccccacc agatccacct gggagagctg
     3421 cacgccattc tgcggcggca ggaagatttt tacccattcc tgaaggacaa ccgggaaaag
     3481 atcgagaaga tcctgacctt ccgcatcccc tactacgtgg gccctctggc caggggaaac
     3541 agcagattcg cctggatgac cagaaagagc gaggaaacca tcaccccctg gaacttcgag
     3601 gaagtggtgg acaagggcgc ttccgcccag agcttcatcg agcggatgac caacttcgat
     3661 aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga gtacttcacc
     3721 gtgtataacg agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc
     3781 ctgagcggcg agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg
     3841 accgtgaagc agctgaaaga ggactacttc aagaaaatcg agtgcttcga ctccgtggaa
     3901 atctccggcg tggaagatcg gttcaacgcc tccctgggca cataccacga tctgctgaaa
     3961 attatcaagg acaaggactt cctggacaat gaggaaaacg aggacattct ggaagatatc
     4021 gtgctgaccc tgacactgtt tgaggacaga gagatgatcg aggaacggct gaaaacctat
     4081 gcccacctgt tcgacgacaa agtgatgaag cagctgaagc ggcggagata caccggctgg
     4141 ggcaggctga gccggaagct gatcaacggc atccgggaca agcagtccgg caagacaatc
     4201 ctggatttcc tgaagtccga cggcttcgcc aacagaaact tcatgcagct gatccacgac
     4261 gacagcctga cctttaaaga ggacatccag aaagcccagg tgtccggcca gggcgatagc
     4321 ctgcacgagc acattgccaa tctggccggc agccccgcca ttaagaaggg catcctgcag
     4381 acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc
     4441 gtgatcgaaa tggccagaga gaaccagacc acccagaagg gacagaagaa cagccgcgag
     4501 agaatgaagc ggatcgaaga gggcatcaaa gagctgggca gccagatcct gaaagaacac
     4561 cccgtggaaa acacccagct gcagaacgag aagctgtacc tgtactacct gcagaatggg
     4621 cgggatatgt acgtggacca ggaactggac atcaaccggc tgtccgacta cgatgtggac
     4681 catatcgtgc ctcagagctt tctgaaggac gactccatcg acaacaaggt gctgaccaga
     4741 agcgacaaga accggggcaa gagcgacaac gtgccctccg aagaggtcgt gaagaagatg
     4801 aagaactact ggcggcagct gctgaacgcc aagctgatta cccagagaaa gttcgacaat
     4861 ctgaccaagg ccgagagagg cggcctgagc gaactggata aggccggctt catcaagaga
     4921 cagctggtgg aaacccggca gatcacaaag cacgtggcac agatcctgga ctcccggatg
     4981 aacactaagt acgacgagaa tgacaagctg atccgggaag tgaaagtgat caccctgaag
     5041 tccaagctgg tgtccgattt ccggaaggat ttccagtttt acaaagtgcg cgagatcaac
     5101 aactaccacc acgcccacga cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa
     5161 aagtacccta agctggaaag cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg
     5221 aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta cttcttctac
     5281 agcaacatca tgaacttttt caagaccgag attaccctgg ccaacggcga gatccggaag
     5341 cggcctctga tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa gggccgggat
     5401 tttgccaccg tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa aaagaccgag
     5461 gtgcagacag gcggcttcag caaagagtct atcctgccca agaggaacag cgataagctg
     5521 atcgccagaa agaaggactg ggaccctaag aagtacggcg gcttcgacag ccccaccgtg
     5581 gcctattctg tgctggtggt ggccaaagtg gaaaagggca agtccaagaa actgaagagt
     5641 gtgaaagagc tgctggggat caccatcatg gaaagaagca gcttcgagaa gaatcccatc
     5701 gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat caagctgcct
     5761 aagtactccc tgttcgagct ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa
     5821 ctgcagaagg gaaacgaact ggccctgccc tccaaatatg tgaacttcct gtacctggcc
     5881 agccactatg agaagctgaa gggctccccc gaggataatg agcagaaaca gctgtttgtg
     5941 gaacagcaca agcactacct ggacgagatc atcgagcaga tcagcgagtt ctccaagaga
     6001 gtgatcctgg ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa gcaccgggat
     6061 aagcccatca gagagcaggc cgagaatatc atccacctgt ttaccctgac caatctggga
     6121 gcccctgccg ccttcaagta ctttgacacc accatcgacc ggaagaggta caccagcacc
     6181 aaagaggtgc tggacgccac cctgatccac cagagcatca ccggcctgta cgagacacgg
     6241 atcgacctgt ctcagctggg aggcgacagc cccaagaaga agagaaaggt ggaggccagc
     6301 taagaattct aactagagct cgctgatcag cctcgactgt gccttctagt tgccagccat
     6361 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     6421 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     6481 ggggtggggt ggggcaggac agcaaggggg aggattggga agagaatagc aggcatgctg
     6541 gggagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     6601 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     6661 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     6721 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     6781 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     6841 cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     6901 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     6961 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     7021 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     7081 aactggaaca acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc
     7141 gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     7201 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     7261 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     7321 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     7381 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     7441 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     7501 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     7561 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     7621 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     7681 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     7741 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     7801 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     7861 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     7921 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     7981 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     8041 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     8101 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     8161 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     8221 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     8281 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtggaagcc gcggtatcat
     8341 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     8401 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     8461 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     8521 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     8581 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     8641 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     8701 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     8761 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     8821 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     8881 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     8941 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     9001 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     9061 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     9121 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     9181 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     9241 cgcggccttt tt
//
