LOCUS       sequence-291889-        9285 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Endogenous tagging of Bassoon: N-terminal (amino acid position: L7).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9285)
  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 9285)
  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-Bsn KI
FEATURES             Location/Qualifiers
     source          1..9285
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        39..287
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     39..59
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     210..229
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        308..383
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             join(525..527,528..1238)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(525..527,528..1238)
                     /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(828..847)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1175..1196
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     enhancer        1327..1612
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        1614..1891
                     /label=chicken beta-actin promoter
     intron          1892..2119
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             2140..2166
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2170..2190
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2200..6300
                     /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             2200..6300
                     /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             6304..6324
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6364..6381)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6370..6577
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6586..6726
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6586..6715
                     /label=AAV2 ITR
     rep_origin      6801..7256
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6888..6907)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7098..7119
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7273..7292)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7392..7414
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7452..7470)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7538..7642
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7643..7711,7712..8503)
                     /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(7861..7880)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8674..9262
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9163..9182
                     /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 aaacaccggg caacgaggcc
      301 agcctgggtt ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg
      361 aaaaagtggc accgagtcgg tgcttttttg ttttagagct agaaatagca agttaaaata
      421 aggctagtcc gtttttagcg cgtgcgccaa ttctgcagac aaatggctct agaagctttc
      481 gaagacccta gaccaccagg ctggcctcgt tgcccgctag cggagtgagc aagggcgagg
      541 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
      601 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
      661 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
      721 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
      781 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
      841 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
      901 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
      961 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     1021 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     1081 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     1141 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     1201 ccgccgggat cactctcggc atggacgagc tgtacaagag cgctgcctcc aggctggcct
     1261 cgttgcccct cggggtcttc tcctcgagga tcctaggacg cgtcctagga cgcgtccaga
     1321 ggtacccgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     1381 cccattgacg tcaatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     1441 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     1501 tattgacgtc aatgacggta aatggcccgc ctggcattgt gcccagtaca tgaccttatg
     1561 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt
     1621 gagccccacg ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta
     1681 tttatttatt ttttaattat tttgtgcagc gatgggggcg gggggggggg gggggcgcgc
     1741 gccaggcggg gcggggcggg gcgaggggcg gggcggggcg aggcggagag gtgcggcggc
     1801 agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg
     1861 gccctataaa aagcgaagcg cgcggcgggc gggagtcgct gcgcgctgcc ttcgccccgt
     1921 gccccgctcc gccgccgcct cgcgccgccc gccccggctc tgactgaccg cgttactccc
     1981 acaggtgagc gggcgggacg gcccttctcc tccgggctgt aattagctga gcaagaggta
     2041 agggtttaag ggatggttgg ttggtggggt attaatgttt aattacctgg agcacctgcc
     2101 tgaaatcact ttttttcagg ttggaccggt gccaccatgt acccatacga tgttccagat
     2161 tacgcttcgc cgaagaaaaa gcgcaaggtc gaagcgtccg acaagaagta cagcatcggc
     2221 ctggacatcg gcaccaactc tgtgggctgg gccgtgatca ccgacgagta caaggtgccc
     2281 agcaagaaat tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa gaacctgatc
     2341 ggagccctgc tgttcgacag cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc
     2401 agaagaagat acaccagacg gaagaaccgg atctgctatc tgcaagagat cttcagcaac
     2461 gagatggcca aggtggacga cagcttcttc cacagactgg aagagtcctt cctggtggaa
     2521 gaggataaga agcacgagcg gcaccccatc ttcggcaaca tcgtggacga ggtggcctac
     2581 cacgagaagt accccaccat ctaccacctg agaaagaaac tggtggacag caccgacaag
     2641 gccgacctgc ggctgatcta tctggccctg gcccacatga tcaagttccg gggccacttc
     2701 ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt catccagctg
     2761 gtgcagacct acaaccagct gttcgaggaa aaccccatca acgccagcgg cgtggacgcc
     2821 aaggccatcc tgtctgccag actgagcaag agcagacggc tggaaaatct gatcgcccag
     2881 ctgcccggcg agaagaagaa tggcctgttc ggaaacctga ttgccctgag cctgggcctg
     2941 acccccaact tcaagagcaa cttcgacctg gccgaggatg ccaaactgca gctgagcaag
     3001 gacacctacg acgacgacct ggacaacctg ctggcccaga tcggcgacca gtacgccgac
     3061 ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc tgagcgacat cctgagagtg
     3121 aacaccgaga tcaccaaggc ccccctgagc gcctctatga tcaagagata cgacgagcac
     3181 caccaggacc tgaccctgct gaaagctctc gtgcggcagc agctgcctga gaagtacaaa
     3241 gagattttct tcgaccagag caagaacggc tacgccggct acattgacgg cggagccagc
     3301 caggaagagt tctacaagtt catcaagccc atcctggaaa agatggacgg caccgaggaa
     3361 ctgctcgtga agctgaacag agaggacctg ctgcggaagc agcggacctt cgacaacggc
     3421 agcatccccc accagatcca cctgggagag ctgcacgcca ttctgcggcg gcaggaagat
     3481 ttttacccat tcctgaagga caaccgggaa aagatcgaga agatcctgac cttccgcatc
     3541 ccctactacg tgggccctct ggccagggga aacagcagat tcgcctggat gaccagaaag
     3601 agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc
     3661 cagagcttca tcgagcggat gaccaacttc gataagaacc tgcccaacga gaaggtgctg
     3721 cccaagcaca gcctgctgta cgagtacttc accgtgtata acgagctgac caaagtgaaa
     3781 tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc
     3841 gtggacctgc tgttcaagac caaccggaaa gtgaccgtga agcagctgaa agaggactac
     3901 ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga tcggttcaac
     3961 gcctccctgg gcacatacca cgatctgctg aaaattatca aggacaagga cttcctggac
     4021 aatgaggaaa acgaggacat tctggaagat atcgtgctga ccctgacact gtttgaggac
     4081 agagagatga tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga caaagtgatg
     4141 aagcagctga agcggcggag atacaccggc tggggcaggc tgagccggaa gctgatcaac
     4201 ggcatccggg acaagcagtc cggcaagaca atcctggatt tcctgaagtc cgacggcttc
     4261 gccaacagaa acttcatgca gctgatccac gacgacagcc tgacctttaa agaggacatc
     4321 cagaaagccc aggtgtccgg ccagggcgat agcctgcacg agcacattgc caatctggcc
     4381 ggcagccccg ccattaagaa gggcatcctg cagacagtga aggtggtgga cgagctcgtg
     4441 aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg aaatggccag agagaaccag
     4501 accacccaga agggacagaa gaacagccgc gagagaatga agcggatcga agagggcatc
     4561 aaagagctgg gcagccagat cctgaaagaa caccccgtgg aaaacaccca gctgcagaac
     4621 gagaagctgt acctgtacta cctgcagaat gggcgggata tgtacgtgga ccaggaactg
     4681 gacatcaacc ggctgtccga ctacgatgtg gaccatatcg tgcctcagag ctttctgaag
     4741 gacgactcca tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg caagagcgac
     4801 aacgtgccct ccgaagaggt cgtgaagaag atgaagaact actggcggca gctgctgaac
     4861 gccaagctga ttacccagag aaagttcgac aatctgacca aggccgagag aggcggcctg
     4921 agcgaactgg ataaggccgg cttcatcaag agacagctgg tggaaacccg gcagatcaca
     4981 aagcacgtgg cacagatcct ggactcccgg atgaacacta agtacgacga gaatgacaag
     5041 ctgatccggg aagtgaaagt gatcaccctg aagtccaagc tggtgtccga tttccggaag
     5101 gatttccagt tttacaaagt gcgcgagatc aacaactacc accacgccca cgacgcctac
     5161 ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga aagcgagttc
     5221 gtgtacggcg actacaaggt gtacgacgtg cggaagatga tcgccaagag cgagcaggaa
     5281 atcggcaagg ctaccgccaa gtacttcttc tacagcaaca tcatgaactt tttcaagacc
     5341 gagattaccc tggccaacgg cgagatccgg aagcggcctc tgatcgagac aaacggcgaa
     5401 accggggaga tcgtgtggga taagggccgg gattttgcca ccgtgcggaa agtgctgagc
     5461 atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt cagcaaagag
     5521 tctatcctgc ccaagaggaa cagcgataag ctgatcgcca gaaagaagga ctgggaccct
     5581 aagaagtacg gcggcttcga cagccccacc gtggcctatt ctgtgctggt ggtggccaaa
     5641 gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag agctgctggg gatcaccatc
     5701 atggaaagaa gcagcttcga gaagaatccc atcgactttc tggaagccaa gggctacaaa
     5761 gaagtgaaaa aggacctgat catcaagctg cctaagtact ccctgttcga gctggaaaac
     5821 ggccggaaga gaatgctggc ctctgccggc gaactgcaga agggaaacga actggccctg
     5881 ccctccaaat atgtgaactt cctgtacctg gccagccact atgagaagct gaagggctcc
     5941 cccgaggata atgagcagaa acagctgttt gtggaacagc acaagcacta cctggacgag
     6001 atcatcgagc agatcagcga gttctccaag agagtgatcc tggccgacgc taatctggac
     6061 aaagtgctgt ccgcctacaa caagcaccgg gataagccca tcagagagca ggccgagaat
     6121 atcatccacc tgtttaccct gaccaatctg ggagcccctg ccgccttcaa gtactttgac
     6181 accaccatcg accggaagag gtacaccagc accaaagagg tgctggacgc caccctgatc
     6241 caccagagca tcaccggcct gtacgagaca cggatcgacc tgtctcagct gggaggcgac
     6301 agccccaaga agaagagaaa ggtggaggcc agctaagaat tctaactaga gctcgctgat
     6361 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     6421 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     6481 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     6541 gggaggattg ggaagagaat agcaggcatg ctggggagcg gccgcaggaa cccctagtga
     6601 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     6661 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     6721 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     6781 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     6841 ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt
     6901 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     6961 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     7021 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     7081 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc
     7141 gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga
     7201 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     7261 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     7321 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     7381 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     7441 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     7501 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     7561 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     7621 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     7681 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     7741 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     7801 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     7861 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     7921 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     7981 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     8041 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     8101 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     8161 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     8221 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     8281 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     8341 tggagccggt gagcgtggaa gccgcggtat cattgcagca ctggggccag atggtaagcc
     8401 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     8461 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     8521 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     8581 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     8641 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     8701 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     8761 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     8821 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     8881 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     8941 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     9001 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     9061 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     9121 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     9181 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     9241 aggggggcgg agcctatgga aaaacgccag caacgcggcc ttttt
//
