LOCUS       sequence-257263-        9254 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Endogenous tagging of Munc13-1: C-terminal (amino acid position: 
            A1762).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9254)
  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 9254)
  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 unc13a GFP KI
FEATURES             Location/Qualifiers
     source          1..9254
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        62..310
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     62..82
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     233..252
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        332..407
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             join(535..537,538..1251)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(535..537,538..1248)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(838..857)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1185..1206
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     enhancer        1319..1604
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        1606..1883
                     /label=chicken beta-actin promoter
     intron          1884..2111
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             2132..2158
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2162..2182
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2192..6292
                     /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             2192..6292
                     /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             6296..6316
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6356..6373)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6362..6569
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6578..6718
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6578..6707
                     /label=AAV2 ITR
     rep_origin      6793..7248
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6880..6899)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7090..7111
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7265..7284)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7384..7406
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7444..7462)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7530..7634
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7635..7703,7704..8495)
                     /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(7853..7872)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8666..9254
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9155..9174
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 tgagggccta tttcccatga ttccttcata tttgcatata cgatacaagg ctgttagaga
      121 gataattgga attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag
      181 aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca
      241 tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg
      301 acgaaacacc gcaaaacgcg cgctagggcg cgttttagag ctagaaatag caagttaaaa
      361 taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt tttgttttag
      421 agctagaaat agcaagttaa aataaggcta gtccgttttt agcgcgtgcg ccaattctgc
      481 agacaaatgg ctctagaagc ttcaaaacgc gcgctagggc gcgggagcgg tggagtgagc
      541 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
      601 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
      661 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
      721 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
      781 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
      841 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
      901 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
      961 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
     1021 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     1081 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     1141 acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     1201 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta acaaaacgcg
     1261 cgctagggcg cgggctcgag gatcctagga cgcgtcctag gacgcgtcca gaggtacccg
     1321 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     1381 cgtcaatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     1441 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     1501 tcaatgacgg taaatggccc gcctggcatt gtgcccagta catgacctta tgggactttc
     1561 ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag gtgagcccca
     1621 cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg tatttattta
     1681 ttttttaatt attttgtgca gcgatggggg cggggggggg gggggggcgc gcgccrgggg
     1741 gggggggggg gggggggggg gggggsgggg sgrggcggag aggtgcggcg gcagccaatc
     1801 agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata
     1861 aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct
     1921 ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga
     1981 gcgggcggga cggcccttct cctccgggct gtaattagct gagcaagagg taagggttta
     2041 agggatggtt ggttggtggg gtattaatgt ttaattacct ggagcacctg cctgaaatca
     2101 ctttttttca ggttggaccg gtgccaccat gtacccatac gatgttccag attacgcttc
     2161 gccgaagaaa aagcgcaagg tcgaagcgtc cgacaagaag tacagcatcg gcctggacat
     2221 cggcaccaac tctgtgggct gggccgtgat caccgacgag tacaaggtgc ccagcaagaa
     2281 attcaaggtg ctgggcaaca ccgaccggca cagcatcaag aagaacctga tcggagccct
     2341 gctgttcgac agcggcgaaa cagccgaggc cacccggctg aagagaaccg ccagaagaag
     2401 atacaccaga cggaagaacc ggatctgcta tctgcaagag atcttcagca acgagatggc
     2461 caaggtggac gacagcttct tccacagact ggaagagtcc ttcctggtgg aagaggataa
     2521 gaagcacgag cggcacccca tcttcggcaa catcgtggac gaggtggcct accacgagaa
     2581 gtaccccacc atctaccacc tgagaaagaa actggtggac agcaccgaca aggccgacct
     2641 gcggctgatc tatctggccc tggcccacat gatcaagttc cggggccact tcctgatcga
     2701 gggcgacctg aaccccgaca acagcgacgt ggacaagctg ttcatccagc tggtgcagac
     2761 ctacaaccag ctgttcgagg aaaaccccat caacgccagc ggcgtggacg ccaaggccat
     2821 cctgtctgcc agactgagca agagcagacg gctggaaaat ctgatcgccc agctgcccgg
     2881 cgagaagaag aatggcctgt tcggaaacct gattgccctg agcctgggcc tgacccccaa
     2941 cttcaagagc aacttcgacc tggccgagga tgccaaactg cagctgagca aggacaccta
     3001 cgacgacgac ctggacaacc tgctggccca gatcggcgac cagtacgccg acctgtttct
     3061 ggccgccaag aacctgtccg acgccatcct gctgagcgac atcctgagag tgaacaccga
     3121 gatcaccaag gcccccctga gcgcctctat gatcaagaga tacgacgagc accaccagga
     3181 cctgaccctg ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca aagagatttt
     3241 cttcgaccag agcaagaacg gctacgccgg ctacattgac ggcggagcca gccaggaaga
     3301 gttctacaag ttcatcaagc ccatcctgga aaagatggac ggcaccgagg aactgctcgt
     3361 gaagctgaac agagaggacc tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc
     3421 ccaccagatc cacctgggag agctgcacgc cattctgcgg cggcaggaag atttttaccc
     3481 attcctgaag gacaaccggg aaaagatcga gaagatcctg accttccgca tcccctacta
     3541 cgtgggccct ctggccaggg gaaacagcag attcgcctgg atgaccagaa agagcgagga
     3601 aaccatcacc ccctggaact tcgaggaagt ggtggacaag ggcgcttccg cccagagctt
     3661 catcgagcgg atgaccaact tcgataagaa cctgcccaac gagaaggtgc tgcccaagca
     3721 cagcctgctg tacgagtact tcaccgtgta taacgagctg accaaagtga aatacgtgac
     3781 cgagggaatg agaaagcccg ccttcctgag cggcgagcag aaaaaggcca tcgtggacct
     3841 gctgttcaag accaaccgga aagtgaccgt gaagcagctg aaagaggact acttcaagaa
     3901 aatcgagtgc ttcgactccg tggaaatctc cggcgtggaa gatcggttca acgcctccct
     3961 gggcacatac cacgatctgc tgaaaattat caaggacaag gacttcctgg acaatgagga
     4021 aaacgaggac attctggaag atatcgtgct gaccctgaca ctgtttgagg acagagagat
     4081 gatcgaggaa cggctgaaaa cctatgccca cctgttcgac gacaaagtga tgaagcagct
     4141 gaagcggcgg agatacaccg gctggggcag gctgagccgg aagctgatca acggcatccg
     4201 ggacaagcag tccggcaaga caatcctgga tttcctgaag tccgacggct tcgccaacag
     4261 aaacttcatg cagctgatcc acgacgacag cctgaccttt aaagaggaca tccagaaagc
     4321 ccaggtgtcc ggccagggcg atagcctgca cgagcacatt gccaatctgg ccggcagccc
     4381 cgccattaag aagggcatcc tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat
     4441 gggccggcac aagcccgaga acatcgtgat cgaaatggcc agagagaacc agaccaccca
     4501 gaagggacag aagaacagcc gcgagagaat gaagcggatc gaagagggca tcaaagagct
     4561 gggcagccag atcctgaaag aacaccccgt ggaaaacacc cagctgcaga acgagaagct
     4621 gtacctgtac tacctgcaga atgggcggga tatgtacgtg gaccaggaac tggacatcaa
     4681 ccggctgtcc gactacgatg tggaccatat cgtgcctcag agctttctga aggacgactc
     4741 catcgacaac aaggtgctga ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc
     4801 ctccgaagag gtcgtgaaga agatgaagaa ctactggcgg cagctgctga acgccaagct
     4861 gattacccag agaaagttcg acaatctgac caaggccgag agaggcggcc tgagcgaact
     4921 ggataaggcc ggcttcatca agagacagct ggtggaaacc cggcagatca caaagcacgt
     4981 ggcacagatc ctggactccc ggatgaacac taagtacgac gagaatgaca agctgatccg
     5041 ggaagtgaaa gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga aggatttcca
     5101 gttttacaaa gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct acctgaacgc
     5161 cgtcgtggga accgccctga tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg
     5221 cgactacaag gtgtacgacg tgcggaagat gatcgccaag agcgagcagg aaatcggcaa
     5281 ggctaccgcc aagtacttct tctacagcaa catcatgaac tttttcaaga ccgagattac
     5341 cctggccaac ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg aaaccgggga
     5401 gatcgtgtgg gataagggcc gggattttgc caccgtgcgg aaagtgctga gcatgcccca
     5461 agtgaatatc gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag agtctatcct
     5521 gcccaagagg aacagcgata agctgatcgc cagaaagaag gactgggacc ctaagaagta
     5581 cggcggcttc gacagcccca ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa
     5641 gggcaagtcc aagaaactga agagtgtgaa agagctgctg gggatcacca tcatggaaag
     5701 aagcagcttc gagaagaatc ccatcgactt tctggaagcc aagggctaca aagaagtgaa
     5761 aaaggacctg atcatcaagc tgcctaagta ctccctgttc gagctggaaa acggccggaa
     5821 gagaatgctg gcctctgccg gcgaactgca gaagggaaac gaactggccc tgccctccaa
     5881 atatgtgaac ttcctgtacc tggccagcca ctatgagaag ctgaagggct cccccgagga
     5941 taatgagcag aaacagctgt ttgtggaaca gcacaagcac tacctggacg agatcatcga
     6001 gcagatcagc gagttctcca agagagtgat cctggccgac gctaatctgg acaaagtgct
     6061 gtccgcctac aacaagcacc gggataagcc catcagagag caggccgaga atatcatcca
     6121 cctgtttacc ctgaccaatc tgggagcccc tgccgccttc aagtactttg acaccaccat
     6181 cgaccggaag aggtacacca gcaccaaaga ggtgctggac gccaccctga tccaccagag
     6241 catcaccggc ctgtacgaga cacggatcga cctgtctcag ctgggaggcg acagccccaa
     6301 gaagaagaga aaggtggagg ccagctaaga attctaacta gagctcgctg atcagcctcg
     6361 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     6421 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     6481 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     6541 tgggaagaga atagcaggca tgctggggag cggccgcagg aacccctagt gatggagttg
     6601 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     6661 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg
     6721 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     6781 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     6841 gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt
     6901 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     6961 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     7021 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     7081 tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt
     7141 cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt
     7201 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     7261 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     7321 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     7381 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     7441 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     7501 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     7561 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     7621 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     7681 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     7741 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     7801 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     7861 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     7921 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     7981 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     8041 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     8101 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     8161 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     8221 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     8281 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     8341 gtgagcgtgg aagccgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     8401 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     8461 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     8521 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     8581 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     8641 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     8701 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     8761 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     8821 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     8881 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     8941 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     9001 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     9061 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     9121 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     9181 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     9241 ggagcctatg gaaa
//
