LOCUS       sequence-257135-        9320 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Endogenous tagging of Tarp-gamma-8: Intramolecular (amino acid 
            position: A408).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9320)
  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 9320)
  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 CACNG8-GFP KI #2
FEATURES             Location/Qualifiers
     source          1..9320
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        107..392
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        394..671
                     /label=chicken beta-actin promoter
     intron          672..899
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             920..946
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             950..970
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             980..5080
                     /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             980..5080
                     /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             5084..5104
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(5144..5161)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5150..5357
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5366..5506
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5366..5495
                     /label=AAV2 ITR
     rep_origin      5581..6036
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5668..5687)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5878..5899
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6053..6072)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6172..6194
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6232..6250)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6318..6422
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6423..6491,6492..7283)
                     /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(6641..6660)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7454..8042
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7943..7962
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8104..8352
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8104..8124
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8275..8294
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8374..8449
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             join(8601..8603,8604..9311)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELY"
     CDS             join(8601..8603,8604..9311)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELY"
     primer_bind     complement(8904..8923)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     9251..9272
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gctggcgctg gagcgggtgc aggggctggc agcgctggcg tgttggtgtt ggacgcggcg
       61 gactcgagga tcctaggacg cgtcctagga cgcgtccaga ggtacccgtt acataactta
      121 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaatagtaa
      181 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
      241 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
      301 aatggcccgc ctggcattgt gcccagtaca tgaccttatg ggactttcct acttggcagt
      361 acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg ttctgcttca
      421 ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt ttttaattat
      481 tttgtgcagc gatgggggcg gggggggggg gggggcgcgc gccggggggg gggggggggg
      541 gggggggggg ggggggggsg rggcggagag gtgcggcggc agccaatcag agcggcgcgc
      601 tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg
      661 cgcggcgggc gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct
      721 cgcgccgccc gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg
      781 gcccttctcc tccgggctgt aattagctga gcaagaggta agggtttaag ggatggttgg
      841 ttggtggggt attaatgttt aattacctgg agcacctgcc tgaaatcact ttttttcagg
      901 ttggaccggt gccaccatgt acccatacga tgttccagat tacgcttcgc cgaagaaaaa
      961 gcgcaaggtc gaagcgtccg acaagaagta cagcatcggc ctggacatcg gcaccaactc
     1021 tgtgggctgg gccgtgatca ccgacgagta caaggtgccc agcaagaaat tcaaggtgct
     1081 gggcaacacc gaccggcaca gcatcaagaa gaacctgatc ggagccctgc tgttcgacag
     1141 cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc agaagaagat acaccagacg
     1201 gaagaaccgg atctgctatc tgcaagagat cttcagcaac gagatggcca aggtggacga
     1261 cagcttcttc cacagactgg aagagtcctt cctggtggaa gaggataaga agcacgagcg
     1321 gcaccccatc ttcggcaaca tcgtggacga ggtggcctac cacgagaagt accccaccat
     1381 ctaccacctg agaaagaaac tggtggacag caccgacaag gccgacctgc ggctgatcta
     1441 tctggccctg gcccacatga tcaagttccg gggccacttc ctgatcgagg gcgacctgaa
     1501 ccccgacaac agcgacgtgg acaagctgtt catccagctg gtgcagacct acaaccagct
     1561 gttcgaggaa aaccccatca acgccagcgg cgtggacgcc aaggccatcc tgtctgccag
     1621 actgagcaag agcagacggc tggaaaatct gatcgcccag ctgcccggcg agaagaagaa
     1681 tggcctgttc ggaaacctga ttgccctgag cctgggcctg acccccaact tcaagagcaa
     1741 cttcgacctg gccgaggatg ccaaactgca gctgagcaag gacacctacg acgacgacct
     1801 ggacaacctg ctggcccaga tcggcgacca gtacgccgac ctgtttctgg ccgccaagaa
     1861 cctgtccgac gccatcctgc tgagcgacat cctgagagtg aacaccgaga tcaccaaggc
     1921 ccccctgagc gcctctatga tcaagagata cgacgagcac caccaggacc tgaccctgct
     1981 gaaagctctc gtgcggcagc agctgcctga gaagtacaaa gagattttct tcgaccagag
     2041 caagaacggc tacgccggct acattgacgg cggagccagc caggaagagt tctacaagtt
     2101 catcaagccc atcctggaaa agatggacgg caccgaggaa ctgctcgtga agctgaacag
     2161 agaggacctg ctgcggaagc agcggacctt cgacaacggc agcatccccc accagatcca
     2221 cctgggagag ctgcacgcca ttctgcggcg gcaggaagat ttttacccat tcctgaagga
     2281 caaccgggaa aagatcgaga agatcctgac cttccgcatc ccctactacg tgggccctct
     2341 ggccagggga aacagcagat tcgcctggat gaccagaaag agcgaggaaa ccatcacccc
     2401 ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc cagagcttca tcgagcggat
     2461 gaccaacttc gataagaacc tgcccaacga gaaggtgctg cccaagcaca gcctgctgta
     2521 cgagtacttc accgtgtata acgagctgac caaagtgaaa tacgtgaccg agggaatgag
     2581 aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc tgttcaagac
     2641 caaccggaaa gtgaccgtga agcagctgaa agaggactac ttcaagaaaa tcgagtgctt
     2701 cgactccgtg gaaatctccg gcgtggaaga tcggttcaac gcctccctgg gcacatacca
     2761 cgatctgctg aaaattatca aggacaagga cttcctggac aatgaggaaa acgaggacat
     2821 tctggaagat atcgtgctga ccctgacact gtttgaggac agagagatga tcgaggaacg
     2881 gctgaaaacc tatgcccacc tgttcgacga caaagtgatg aagcagctga agcggcggag
     2941 atacaccggc tggggcaggc tgagccggaa gctgatcaac ggcatccggg acaagcagtc
     3001 cggcaagaca atcctggatt tcctgaagtc cgacggcttc gccaacagaa acttcatgca
     3061 gctgatccac gacgacagcc tgacctttaa agaggacatc cagaaagccc aggtgtccgg
     3121 ccagggcgat agcctgcacg agcacattgc caatctggcc ggcagccccg ccattaagaa
     3181 gggcatcctg cagacagtga aggtggtgga cgagctcgtg aaagtgatgg gccggcacaa
     3241 gcccgagaac atcgtgatcg aaatggccag agagaaccag accacccaga agggacagaa
     3301 gaacagccgc gagagaatga agcggatcga agagggcatc aaagagctgg gcagccagat
     3361 cctgaaagaa caccccgtgg aaaacaccca gctgcagaac gagaagctgt acctgtacta
     3421 cctgcagaat gggcgggata tgtacgtgga ccaggaactg gacatcaacc ggctgtccga
     3481 ctacgatgtg gaccatatcg tgcctcagag ctttctgaag gacgactcca tcgacaacaa
     3541 ggtgctgacc agaagcgaca agaaccgggg caagagcgac aacgtgccct ccgaagaggt
     3601 cgtgaagaag atgaagaact actggcggca gctgctgaac gccaagctga ttacccagag
     3661 aaagttcgac aatctgacca aggccgagag aggcggcctg agcgaactgg ataaggccgg
     3721 cttcatcaag agacagctgg tggaaacccg gcagatcaca aagcacgtgg cacagatcct
     3781 ggactcccgg atgaacacta agtacgacga gaatgacaag ctgatccggg aagtgaaagt
     3841 gatcaccctg aagtccaagc tggtgtccga tttccggaag gatttccagt tttacaaagt
     3901 gcgcgagatc aacaactacc accacgccca cgacgcctac ctgaacgccg tcgtgggaac
     3961 cgccctgatc aaaaagtacc ctaagctgga aagcgagttc gtgtacggcg actacaaggt
     4021 gtacgacgtg cggaagatga tcgccaagag cgagcaggaa atcggcaagg ctaccgccaa
     4081 gtacttcttc tacagcaaca tcatgaactt tttcaagacc gagattaccc tggccaacgg
     4141 cgagatccgg aagcggcctc tgatcgagac aaacggcgaa accggggaga tcgtgtggga
     4201 taagggccgg gattttgcca ccgtgcggaa agtgctgagc atgccccaag tgaatatcgt
     4261 gaaaaagacc gaggtgcaga caggcggctt cagcaaagag tctatcctgc ccaagaggaa
     4321 cagcgataag ctgatcgcca gaaagaagga ctgggaccct aagaagtacg gcggcttcga
     4381 cagccccacc gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg gcaagtccaa
     4441 gaaactgaag agtgtgaaag agctgctggg gatcaccatc atggaaagaa gcagcttcga
     4501 gaagaatccc atcgactttc tggaagccaa gggctacaaa gaagtgaaaa aggacctgat
     4561 catcaagctg cctaagtact ccctgttcga gctggaaaac ggccggaaga gaatgctggc
     4621 ctctgccggc gaactgcaga agggaaacga actggccctg ccctccaaat atgtgaactt
     4681 cctgtacctg gccagccact atgagaagct gaagggctcc cccgaggata atgagcagaa
     4741 acagctgttt gtggaacagc acaagcacta cctggacgag atcatcgagc agatcagcga
     4801 gttctccaag agagtgatcc tggccgacgc taatctggac aaagtgctgt ccgcctacaa
     4861 caagcaccgg gataagccca tcagagagca ggccgagaat atcatccacc tgtttaccct
     4921 gaccaatctg ggagcccctg ccgccttcaa gtactttgac accaccatcg accggaagag
     4981 gtacaccagc accaaagagg tgctggacgc caccctgatc caccagagca tcaccggcct
     5041 gtacgagaca cggatcgacc tgtctcagct gggaggcgac agccccaaga agaagagaaa
     5101 ggtggaggcc agctaagaat tctaactaga gctcgctgat cagcctcgac tgtgccttct
     5161 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     5221 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     5281 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagagaat
     5341 agcaggcatg ctggggagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     5401 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     5461 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     5521 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     5581 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     5641 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     5701 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     5761 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     5821 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     5881 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     5941 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     6001 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     6061 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     6121 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     6181 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     6241 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     6301 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     6361 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     6421 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     6481 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     6541 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     6601 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     6661 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     6721 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     6781 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     6841 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     6901 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     6961 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     7021 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     7081 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtggaa
     7141 gccgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     7201 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     7261 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     7321 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     7381 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     7441 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     7501 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     7561 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     7621 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     7681 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     7741 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     7801 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     7861 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     7921 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     7981 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     8041 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     8101 tgtgagggcc tatttcccat gattccttca tatttgcata tacgatacaa ggctgttaga
     8161 gagataattg gaattaattt gactgtaaac acaaagatat tagtacaaaa tacgtgacgt
     8221 agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa aatggactat
     8281 catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat cttgtggaaa
     8341 ggacgaaaca ccgcgtgttg gtgttggacg cgggttttag agctagaaat agcaagttaa
     8401 aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct tttttgtttt
     8461 agagctagaa atagcaagtt aaaataaggc tagtccgttt ttagcgcgtg cgccaattct
     8521 gcagacaaat ggctctagaa gcttgcgtgt tggtgttgga cgcggcggct agcgcaggta
     8581 gcggctccag tagatctggg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     8641 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     8701 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     8761 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     8821 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     8881 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     8941 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     9001 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     9061 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     9121 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     9181 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
     9241 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     9301 acgagctgta ctctagggga
//
