LOCUS       sequence-291898-        9264 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Endogenous tagging of CaV-beta-1: N-terminal (amino acid position: 
            S8).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9264)
  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 9264)
  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-Cacnb1 KI
FEATURES             Location/Qualifiers
     source          1..9264
                     /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(514..516,517..1227)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(514..516,517..1227)
                     /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(817..836)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1164..1185
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     enhancer        1306..1591
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        1593..1870
                     /label=chicken beta-actin promoter
     intron          1871..2098
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             2119..2145
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2149..2169
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2179..6279
                     /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             2179..6279
                     /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             6283..6303
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6343..6360)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6349..6556
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6565..6705
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6565..6694
                     /label=AAV2 ITR
     rep_origin      6780..7235
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6867..6886)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7077..7098
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7252..7271)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7371..7393
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7431..7449)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7517..7621
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7622..7690,7691..8482)
                     /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(7840..7859)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8653..9241
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9142..9161
                     /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 aaacaccgca gaagagcggc
      301 atgtcccggt tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt
      361 gaaaaagtgg caccgagtcg gtgctttttt gttttagagc tagaaatagc aagttaaaat
      421 aaggctagtc cgtttttagc gcgtgcgcca attctgcaga caaatggctc tagaagcttc
      481 cccgggacat gccgctcttc tggagctagc ggagtgagca agggcgagga gctgttcacc
      541 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
      601 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
      661 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
      721 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
      781 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
      841 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
      901 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
      961 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     1021 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     1081 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
     1141 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     1201 actctcggca tggacgagct gtacaagggc agcgctggcc gcgggacatg ccgctcttct
     1261 gctcgaggat cctaggacgc gtcctaggac gcgtccagag gtacccgtta cataacttac
     1321 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caatagtaac
     1381 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     1441 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     1501 atggcccgcc tggcattgtg cccagtacat gaccttatgg gactttccta cttggcagta
     1561 catctacgta ttagtcatcg ctattaccat ggtcgaggtg agccccacgt tctgcttcac
     1621 tctccccatc tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt
     1681 ttgtgcagcg atgggggcgg gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg
     1741 cgaggggcgg ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct
     1801 ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc
     1861 gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc
     1921 gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg
     1981 cccttctcct ccgggctgta attagctgag caagaggtaa gggtttaagg gatggttggt
     2041 tggtggggta ttaatgttta attacctgga gcacctgcct gaaatcactt tttttcaggt
     2101 tggaccggtg ccaccatgta cccatacgat gttccagatt acgcttcgcc gaagaaaaag
     2161 cgcaaggtcg aagcgtccga caagaagtac agcatcggcc tggacatcgg caccaactct
     2221 gtgggctggg ccgtgatcac cgacgagtac aaggtgccca gcaagaaatt caaggtgctg
     2281 ggcaacaccg accggcacag catcaagaag aacctgatcg gagccctgct gttcgacagc
     2341 ggcgaaacag ccgaggccac ccggctgaag agaaccgcca gaagaagata caccagacgg
     2401 aagaaccgga tctgctatct gcaagagatc ttcagcaacg agatggccaa ggtggacgac
     2461 agcttcttcc acagactgga agagtccttc ctggtggaag aggataagaa gcacgagcgg
     2521 caccccatct tcggcaacat cgtggacgag gtggcctacc acgagaagta ccccaccatc
     2581 taccacctga gaaagaaact ggtggacagc accgacaagg ccgacctgcg gctgatctat
     2641 ctggccctgg cccacatgat caagttccgg ggccacttcc tgatcgaggg cgacctgaac
     2701 cccgacaaca gcgacgtgga caagctgttc atccagctgg tgcagaccta caaccagctg
     2761 ttcgaggaaa accccatcaa cgccagcggc gtggacgcca aggccatcct gtctgccaga
     2821 ctgagcaaga gcagacggct ggaaaatctg atcgcccagc tgcccggcga gaagaagaat
     2881 ggcctgttcg gaaacctgat tgccctgagc ctgggcctga cccccaactt caagagcaac
     2941 ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg acacctacga cgacgacctg
     3001 gacaacctgc tggcccagat cggcgaccag tacgccgacc tgtttctggc cgccaagaac
     3061 ctgtccgacg ccatcctgct gagcgacatc ctgagagtga acaccgagat caccaaggcc
     3121 cccctgagcg cctctatgat caagagatac gacgagcacc accaggacct gaccctgctg
     3181 aaagctctcg tgcggcagca gctgcctgag aagtacaaag agattttctt cgaccagagc
     3241 aagaacggct acgccggcta cattgacggc ggagccagcc aggaagagtt ctacaagttc
     3301 atcaagccca tcctggaaaa gatggacggc accgaggaac tgctcgtgaa gctgaacaga
     3361 gaggacctgc tgcggaagca gcggaccttc gacaacggca gcatccccca ccagatccac
     3421 ctgggagagc tgcacgccat tctgcggcgg caggaagatt tttacccatt cctgaaggac
     3481 aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc cctactacgt gggccctctg
     3541 gccaggggaa acagcagatt cgcctggatg accagaaaga gcgaggaaac catcaccccc
     3601 tggaacttcg aggaagtggt ggacaagggc gcttccgccc agagcttcat cgagcggatg
     3661 accaacttcg ataagaacct gcccaacgag aaggtgctgc ccaagcacag cctgctgtac
     3721 gagtacttca ccgtgtataa cgagctgacc aaagtgaaat acgtgaccga gggaatgaga
     3781 aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg tggacctgct gttcaagacc
     3841 aaccggaaag tgaccgtgaa gcagctgaaa gaggactact tcaagaaaat cgagtgcttc
     3901 gactccgtgg aaatctccgg cgtggaagat cggttcaacg cctccctggg cacataccac
     3961 gatctgctga aaattatcaa ggacaaggac ttcctggaca atgaggaaaa cgaggacatt
     4021 ctggaagata tcgtgctgac cctgacactg tttgaggaca gagagatgat cgaggaacgg
     4081 ctgaaaacct atgcccacct gttcgacgac aaagtgatga agcagctgaa gcggcggaga
     4141 tacaccggct ggggcaggct gagccggaag ctgatcaacg gcatccggga caagcagtcc
     4201 ggcaagacaa tcctggattt cctgaagtcc gacggcttcg ccaacagaaa cttcatgcag
     4261 ctgatccacg acgacagcct gacctttaaa gaggacatcc agaaagccca ggtgtccggc
     4321 cagggcgata gcctgcacga gcacattgcc aatctggccg gcagccccgc cattaagaag
     4381 ggcatcctgc agacagtgaa ggtggtggac gagctcgtga aagtgatggg ccggcacaag
     4441 cccgagaaca tcgtgatcga aatggccaga gagaaccaga ccacccagaa gggacagaag
     4501 aacagccgcg agagaatgaa gcggatcgaa gagggcatca aagagctggg cagccagatc
     4561 ctgaaagaac accccgtgga aaacacccag ctgcagaacg agaagctgta cctgtactac
     4621 ctgcagaatg ggcgggatat gtacgtggac caggaactgg acatcaaccg gctgtccgac
     4681 tacgatgtgg accatatcgt gcctcagagc tttctgaagg acgactccat cgacaacaag
     4741 gtgctgacca gaagcgacaa gaaccggggc aagagcgaca acgtgccctc cgaagaggtc
     4801 gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg ccaagctgat tacccagaga
     4861 aagttcgaca atctgaccaa ggccgagaga ggcggcctga gcgaactgga taaggccggc
     4921 ttcatcaaga gacagctggt ggaaacccgg cagatcacaa agcacgtggc acagatcctg
     4981 gactcccgga tgaacactaa gtacgacgag aatgacaagc tgatccggga agtgaaagtg
     5041 atcaccctga agtccaagct ggtgtccgat ttccggaagg atttccagtt ttacaaagtg
     5101 cgcgagatca acaactacca ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc
     5161 gccctgatca aaaagtaccc taagctggaa agcgagttcg tgtacggcga ctacaaggtg
     5221 tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc taccgccaag
     5281 tacttcttct acagcaacat catgaacttt ttcaagaccg agattaccct ggccaacggc
     5341 gagatccgga agcggcctct gatcgagaca aacggcgaaa ccggggagat cgtgtgggat
     5401 aagggccggg attttgccac cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg
     5461 aaaaagaccg aggtgcagac aggcggcttc agcaaagagt ctatcctgcc caagaggaac
     5521 agcgataagc tgatcgccag aaagaaggac tgggacccta agaagtacgg cggcttcgac
     5581 agccccaccg tggcctattc tgtgctggtg gtggccaaag tggaaaaggg caagtccaag
     5641 aaactgaaga gtgtgaaaga gctgctgggg atcaccatca tggaaagaag cagcttcgag
     5701 aagaatccca tcgactttct ggaagccaag ggctacaaag aagtgaaaaa ggacctgatc
     5761 atcaagctgc ctaagtactc cctgttcgag ctggaaaacg gccggaagag aatgctggcc
     5821 tctgccggcg aactgcagaa gggaaacgaa ctggccctgc cctccaaata tgtgaacttc
     5881 ctgtacctgg ccagccacta tgagaagctg aagggctccc ccgaggataa tgagcagaaa
     5941 cagctgtttg tggaacagca caagcactac ctggacgaga tcatcgagca gatcagcgag
     6001 ttctccaaga gagtgatcct ggccgacgct aatctggaca aagtgctgtc cgcctacaac
     6061 aagcaccggg ataagcccat cagagagcag gccgagaata tcatccacct gtttaccctg
     6121 accaatctgg gagcccctgc cgccttcaag tactttgaca ccaccatcga ccggaagagg
     6181 tacaccagca ccaaagaggt gctggacgcc accctgatcc accagagcat caccggcctg
     6241 tacgagacac ggatcgacct gtctcagctg ggaggcgaca gccccaagaa gaagagaaag
     6301 gtggaggcca gctaagaatt ctaactagag ctcgctgatc agcctcgact gtgccttcta
     6361 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     6421 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     6481 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagagaata
     6541 gcaggcatgc tggggagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     6601 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     6661 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt
     6721 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     6781 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     6841 tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     6901 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     6961 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     7021 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     7081 actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata
     7141 agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     7201 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     7261 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     7321 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     7381 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     7441 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     7501 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     7561 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     7621 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     7681 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     7741 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     7801 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     7861 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     7921 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     7981 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     8041 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     8101 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     8161 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     8221 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     8281 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtggaag
     8341 ccgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     8401 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     8461 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     8521 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     8581 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     8641 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     8701 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     8761 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     8821 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     8881 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     8941 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     9001 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     9061 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     9121 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     9181 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     9241 aaacgccagc aacgcggcct tttt
//
