LOCUS       sequence-360154-        9347 bp    DNA     circular SYN 15-APR-2025
DEFINITION  FlpON knock-in for mRuby3-beta-actin (amino acid position: before 
            start codon).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9347)
  AUTHORS   Droogers WJ, Willems J, MacGillavry HD, de Jong APH
  TITLE     Duplex Labeling and Manipulation of Neuronal Proteins Using 
            Sequential CRISPR/Cas9 Gene Editing.
  JOURNAL   eNeuro. 2022 Jul 18;9(4):ENEURO.0056-22.2022. doi: 
            10.1523/ENEURO.0056-22.2022.
  PUBMED    35851300
REFERENCE   2  (bases 1 to 9347)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1523/ENEURO.0056-22"; journalName: "eNeuro"; date: "2022-07-18- 
            18"; volume: "9"; issue: "4"; pages: "ENEURO.0056-22.2022"
COMMENT     Sequence Label: pOC4 Actb-mRuby3 KI
FEATURES             Location/Qualifiers
     source          1..9347
                     /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"
     protein_bind    348..381
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    420..453
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             655..1365
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby2, with improved brightness and photostability (Bajar 
                     et al., 2016)"
                     /label=mRuby3
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGVQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVTQDTSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTD
                     IALKVDGGGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAV
                     AKYSNLGGGMDELYK"
     enhancer        1415..1700
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        1702..1979
                     /label=chicken beta-actin promoter
     intron          1980..2207
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      2219..2228
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2228..2251
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2255..2275
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2285..6385
                     /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             2285..6385
                     /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             6389..6409
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6449..6466)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6455..6662
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6671..6811
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6671..6800
                     /label=AAV2 ITR
     rep_origin      6886..7341
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6973..6992)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7183..7204
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7358..7377)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7477..7499
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7537..7555)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7623..7727
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7728..7796,7797..8588)
                     /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(7946..7965)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8759..9347
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9248..9267
                     /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 tgtgccttga tagttcgcca gtttaagagc tatgctggaa gttcctattc
      361 tctagaaagt ataggaactt cttttttttt tgcggccgcc cagaattctg gcctgcaggg
      421 aagttcctat tctctagaaa gtataggaac ttccagcata gcaagtttaa ataaggctag
      481 tccgttatca acttgaaaaa gtggcaccga gtcggtgctt ttttgtttta gagctagaaa
      541 tagcaagtta aaataaggct agtccgtttt tagcgcgtgc gccaattctg cagacaaatg
      601 gctctagaag cttccctggc gaactatcaa ggcacattcg ccatggctag cggaatggtg
      661 tctaagggcg aagagctgat caaggaaaat atgcgtatga aggtggtcat ggaaggttcg
      721 gtcaacggcc accaattcaa atgcacaggt gaaggagaag gcagaccgta cgagggagtg
      781 caaaccatga ggatcaaagt catcgaggga ggacccctgc catttgcctt tgacattctt
      841 gccacgtcgt tcatgtatgg cagccgtacc tttatcaagt acccggccga catccctgat
      901 ttctttaaac agtcctttcc tgagggtttt acttgggaaa gagttacgag atacgaagat
      961 ggtggagtcg tcaccgtcac gcaggacacc agccttgagg atggcgagct cgtctacaac
     1021 gtcaaggtca gaggggtaaa ctttccctcc aatggtcccg tgatgcagaa gaagaccaag
     1081 ggttgggagc ctaatacaga gatgatgtat ccagcagatg gtggtctgag aggatacact
     1141 gacatcgcac tgaaagttga tggtggtggc catctgcact gcaacttcgt gacaacttac
     1201 aggtcaaaaa agaccgtcgg gaacatcaag atgcccggtg tccatgccgt tgatcaccgc
     1261 ctggaaagga tcgaggagag tgacaatgaa acctacgtag tgcaaagaga agtggcagtt
     1321 gccaaataca gcaaccttgg tggtggcatg gacgagctgt acaagggcag cgctccctgg
     1381 cgaactatca aggcacaacg cgtccagagg tacccgttac ataacttacg gtaaatggcc
     1441 cgcctggctg accgcccaac gacccccgcc cattgacgtc aatagtaacg ccaataggga
     1501 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     1561 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     1621 ggcattgtgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     1681 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
     1741 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
     1801 tgggggcggg gggggggggg gggcgcgcgc crggsggggs ggggsggggs grggggsggg
     1861 gsggggsgrg gcggagaggk gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc
     1921 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
     1981 gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc
     2041 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
     2101 cgggctgtaa ttagctgagc aagaggtaag ggtttaaggg atggttggtt ggtggggtat
     2161 taatgtttaa ttacctggag cacctgcctg aaatcacttt ttttcaggtt ggaccggtgc
     2221 caccatggat tacaaagatg atgacgataa atcgccgaag aaaaagcgca aggtcgaagc
     2281 gtccgacaag aagtacagca tcggcctgga catcggcacc aactctgtgg gctgggccgt
     2341 gatcaccgac gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg
     2401 gcacagcatc aagaagaacc tgatcggagc cctgctgttc gacagcggcg aaacagccga
     2461 ggccacccgg ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg
     2521 ctatctgcaa gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag
     2581 actggaagag tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg
     2641 caacatcgtg gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa
     2701 gaaactggtg gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca
     2761 catgatcaag ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga
     2821 cgtggacaag ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc
     2881 catcaacgcc agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag
     2941 acggctggaa aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggaaa
     3001 cctgattgcc ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga
     3061 ggatgccaaa ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc
     3121 ccagatcggc gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat
     3181 cctgctgagc gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc
     3241 tatgatcaag agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg
     3301 gcagcagctg cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc
     3361 cggctacatt gacggcggag ccagccagga agagttctac aagttcatca agcccatcct
     3421 ggaaaagatg gacggcaccg aggaactgct cgtgaagctg aacagagagg acctgctgcg
     3481 gaagcagcgg accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca
     3541 cgccattctg cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat
     3601 cgagaagatc ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag
     3661 cagattcgcc tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga
     3721 agtggtggac aagggcgctt ccgcccagag cttcatcgag cggatgacca acttcgataa
     3781 gaacctgccc aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt
     3841 gtataacgag ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct
     3901 gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac
     3961 cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat
     4021 ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat
     4081 tatcaaggac aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt
     4141 gctgaccctg acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc
     4201 ccacctgttc gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg
     4261 caggctgagc cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct
     4321 ggatttcctg aagtccgacg gcttcgccaa cagaaacttc atgcagctga tccacgacga
     4381 cagcctgacc tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct
     4441 gcacgagcac attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac
     4501 agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt
     4561 gatcgaaatg gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag
     4621 aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc
     4681 cgtggaaaac acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg
     4741 ggatatgtac gtggaccagg aactggacat caaccggctg tccgactacg atgtggacca
     4801 tatcgtgcct cagagctttc tgaaggacga ctccatcgac aacaaggtgc tgaccagaag
     4861 cgacaagaac cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa
     4921 gaactactgg cggcagctgc tgaacgccaa gctgattacc cagagaaagt tcgacaatct
     4981 gaccaaggcc gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca
     5041 gctggtggaa acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa
     5101 cactaagtac gacgagaatg acaagctgat ccgggaagtg aaagtgatca ccctgaagtc
     5161 caagctggtg tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa
     5221 ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa
     5281 gtaccctaag ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa
     5341 gatgatcgcc aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag
     5401 caacatcatg aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg
     5461 gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg gccgggattt
     5521 tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt
     5581 gcagacaggc ggcttcagca aagagtctat cctgcccaag aggaacagcg ataagctgat
     5641 cgccagaaag aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc
     5701 ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt
     5761 gaaagagctg ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga
     5821 ctttctggaa gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa
     5881 gtactccctg ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgaact
     5941 gcagaaggga aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag
     6001 ccactatgag aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga
     6061 acagcacaag cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt
     6121 gatcctggcc gacgctaatc tggacaaagt gctgtccgcc tacaacaagc accgggataa
     6181 gcccatcaga gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc
     6241 ccctgccgcc ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa
     6301 agaggtgctg gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacggat
     6361 cgacctgtct cagctgggag gcgacagccc caagaagaag agaaaggtgg aggccagcta
     6421 agaattctaa ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     6481 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     6541 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     6601 ggtggggtgg ggcaggacag caagggggag gattgggaag agaatagcag gcatgctggg
     6661 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     6721 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     6781 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg
     6841 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
     6901 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     6961 ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     7021 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     7081 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     7141 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     7201 ctggaacaac actcaactct atctcgggct attcttttga tttataaggg attttgccga
     7261 tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     7321 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
     7381 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     7441 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     7501 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     7561 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     7621 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     7681 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     7741 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     7801 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     7861 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     7921 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     7981 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     8041 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     8101 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     8161 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     8221 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     8281 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     8341 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     8401 ctggctggtt tattgctgat aaatctggag ccggtgagcg tggaagccgc ggtatcattg
     8461 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     8521 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     8581 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     8641 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     8701 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     8761 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     8821 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     8881 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     8941 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     9001 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     9061 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     9121 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     9181 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     9241 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     9301 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaa
//
