LOCUS       sequence-257169-        9505 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Endogenous tagging of PSD95: C-terminal (amino acid position: R721).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9505)
  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 9505)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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 Dlg4-GFP KI
FEATURES             Location/Qualifiers
     source          1..9505
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          280..507
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             528..554
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             558..578
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             588..4688
                     /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             588..4688
                     /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             4692..4712
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4752..4769)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4758..4965
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4974..5114
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4974..5103
                     /label=AAV2 ITR
     rep_origin      5189..5644
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5276..5295)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5486..5507
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5661..5680)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5780..5802
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5840..5858)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5926..6030
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6031..6099,6100..6891)
                     /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(6249..6268)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7062..7650
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7551..7570
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        7712..7960
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7712..7732
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7883..7902
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        7982..8057
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             join(8185..8187,8188..8898)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(8185..8187,8188..8898)
                     /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(8488..8507)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8835..8856
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     enhancer        8973..9258
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg
       61 gggggggggg gggggggggg gggggggggg gggngggggg gggggggggg gggggggggg
      121 gggsgsgggn nggggggggg gggggggggs gggggggggg gcggggcgag gcggagaggt
      181 gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg
      241 cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt
      301 cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc cccggctctg actgaccgcg
      361 ttactcccac aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa ttagctgagc
      421 aagaggtaag ggtttaaggg atggttggtt ggtggggtat taatgtttaa ttacctggag
      481 cacctgcctg aaatcacttt ttttcaggtt ggaccggtgc caccatgtac ccatacgatg
      541 ttccagatta cgcttcgccg aagaaaaagc gcaaggtcga agcgtccgac aagaagtaca
      601 gcatcggcct ggacatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca
      661 aggtgcccag caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga
      721 acctgatcgg agccctgctg ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga
      781 gaaccgccag aagaagatac accagacgga agaaccggat ctgctatctg caagagatct
      841 tcagcaacga gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc
      901 tggtggaaga ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg
      961 tggcctacca cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca
     1021 ccgacaaggc cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg
     1081 gccacttcct gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca
     1141 tccagctggt gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg
     1201 tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga
     1261 tcgcccagct gcccggcgag aagaagaatg gcctgttcgg aaacctgatt gccctgagcc
     1321 tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc
     1381 tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt
     1441 acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc
     1501 tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg
     1561 acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga
     1621 agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac attgacggcg
     1681 gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca
     1741 ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg
     1801 acaacggcag catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc
     1861 aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct
     1921 tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga
     1981 ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg
     2041 cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga
     2101 aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac gagctgacca
     2161 aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa
     2221 aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag
     2281 aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc
     2341 ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact
     2401 tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt
     2461 ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca
     2521 aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc
     2581 tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg
     2641 acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag
     2701 aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca
     2761 atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg
     2821 agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag
     2881 agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag
     2941 agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc
     3001 tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc
     3061 aggaactgga catcaaccgg ctgtccgact acgatgtgga ccatatcgtg cctcagagct
     3121 ttctgaagga cgactccatc gacaacaagg tgctgaccag aagcgacaag aaccggggca
     3181 agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcggcagc
     3241 tgctgaacgc caagctgatt acccagagaa agttcgacaa tctgaccaag gccgagagag
     3301 gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc
     3361 agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga
     3421 atgacaagct gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt
     3481 tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg
     3541 acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa
     3601 gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg
     3661 agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt
     3721 tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa
     3781 acggcgaaac cggggagatc gtgtgggata agggccggga ttttgccacc gtgcggaaag
     3841 tgctgagcat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca
     3901 gcaaagagtc tatcctgccc aagaggaaca gcgataagct gatcgccaga aagaaggact
     3961 gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct gtgctggtgg
     4021 tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctgctgggga
     4081 tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg
     4141 gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc
     4201 tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag ggaaacgaac
     4261 tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga
     4321 agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac aagcactacc
     4381 tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta
     4441 atctggacaa agtgctgtcc gcctacaaca agcaccggga taagcccatc agagagcagg
     4501 ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt
     4561 actttgacac caccatcgac cggaagaggt acaccagcac caaagaggtg ctggacgcca
     4621 ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg
     4681 gaggcgacag ccccaagaag aagagaaagg tggaggccag ctaagaattc taactagagc
     4741 tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     4801 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     4861 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     4921 cagcaagggg gaggattggg aagagaatag caggcatgct ggggagcggc cgcaggaacc
     4981 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     5041 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     5101 cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     5161 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
     5221 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct
     5281 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     5341 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     5401 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     5461 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     5521 tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta
     5581 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     5641 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     5701 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     5761 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     5821 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     5881 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5941 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     6001 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     6061 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     6121 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     6181 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     6241 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     6301 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     6361 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     6421 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     6481 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     6541 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     6601 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     6661 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     6721 gataaatctg gagccggtga gcgtggaagc cgcggtatca ttgcagcact ggggccagat
     6781 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6841 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     6901 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     6961 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     7021 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     7081 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     7141 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     7201 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     7261 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     7321 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     7381 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     7441 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     7501 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     7561 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     7621 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     7681 ctggcctttt gctggccttt tgctcacatg tgagggccta tttcccatga ttccttcata
     7741 tttgcatata cgatacaagg ctgttagaga gataattgga attaatttga ctgtaaacac
     7801 aaagatatta gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt
     7861 tttaaaatta tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga
     7921 tttcttggct ttatatatct tgtggaaagg acgaaacacc gaatcagagt ctctctcggg
     7981 cgttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag
     8041 tggcaccgag tcggtgcttt tttgttttag agctagaaat agcaagttaa aataaggcta
     8101 gtccgttttt agcgcgtgcg ccaattctgc agacaaatgg ctctagaagc ttaatcagag
     8161 tctctctcgg gcgggagcgg tggagtgagc aagggcgagg agctgttcac cggggtggtg
     8221 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     8281 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     8341 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     8401 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     8461 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     8521 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     8581 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     8641 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     8701 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     8761 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     8821 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     8881 atggacgagc tgtacaaggg ttcaaaatca gagtctctct cgggcgggct cgaggatcct
     8941 aggacgcgtc ctaggacgcg tccagaggta cccgttacat aacttacggt aaatggcccg
     9001 cctggctgac cgcccaacga cccccgccca ttgacgtcaa tagtaacgcc aatagggact
     9061 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     9121 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     9181 cattgtgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     9241 gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc
     9301 cccccctccc cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg
     9361 ggggcggggg gggggggggg gggsgccgcc gggggggggg gggggggggg gggggggggg
     9421 gggggggggg gggggggggg gggggggggg sgangggggg gggggggggg gggggkgggg
     9481 kkkggggggg gggggggggg ggggg
//
