LOCUS       sequence-257669-        9275 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Endogenous tagging of RIM1: C-terminal (amino acid position: S1615).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9275)
  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 9275)
  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 Rims1-GFP KI
FEATURES             Location/Qualifiers
     source          1..9275
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          159..386
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             407..433
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             437..457
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             467..4567
                     /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             467..4567
                     /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             4571..4591
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4631..4648)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4637..4844
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4853..4993
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4853..4982
                     /label=AAV2 ITR
     rep_origin      5068..5523
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5155..5174)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5365..5386
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5540..5559)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5659..5681
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5719..5737)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5805..5909
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5910..5978,5979..6770)
                     /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(6128..6147)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6941..7529
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7430..7449
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        7591..7839
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7591..7611
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7762..7781
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        7860..7935
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             join(8088..8090,8091..8804)
                     /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(8088..8090,8091..8801)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(8391..8410)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8738..8759
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     enhancer        8877..9162
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        join(9164..9275,1..158)
                     /label=chicken beta-actin promoter
ORIGIN
        1 ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg cggggcgagg cggagaggtg
       61 cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc
      121 ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc
      181 gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt
      241 tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagctgagca
      301 agaggtaagg gtttaaggga tggttggttg gtggggtatt aatgtttaat tacctggagc
      361 acctgcctga aatcactttt tttcaggttg gaccggtgcc accatgtacc catacgatgt
      421 tccagattac gcttcgccga agaaaaagcg caaggtcgaa gcgtccgaca agaagtacag
      481 catcggcctg gacatcggca ccaactctgt gggctgggcc gtgatcaccg acgagtacaa
      541 ggtgcccagc aagaaattca aggtgctggg caacaccgac cggcacagca tcaagaagaa
      601 cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag
      661 aaccgccaga agaagataca ccagacggaa gaaccggatc tgctatctgc aagagatctt
      721 cagcaacgag atggccaagg tggacgacag cttcttccac agactggaag agtccttcct
      781 ggtggaagag gataagaagc acgagcggca ccccatcttc ggcaacatcg tggacgaggt
      841 ggcctaccac gagaagtacc ccaccatcta ccacctgaga aagaaactgg tggacagcac
      901 cgacaaggcc gacctgcggc tgatctatct ggccctggcc cacatgatca agttccgggg
      961 ccacttcctg atcgagggcg acctgaaccc cgacaacagc gacgtggaca agctgttcat
     1021 ccagctggtg cagacctaca accagctgtt cgaggaaaac cccatcaacg ccagcggcgt
     1081 ggacgccaag gccatcctgt ctgccagact gagcaagagc agacggctgg aaaatctgat
     1141 cgcccagctg cccggcgaga agaagaatgg cctgttcgga aacctgattg ccctgagcct
     1201 gggcctgacc cccaacttca agagcaactt cgacctggcc gaggatgcca aactgcagct
     1261 gagcaaggac acctacgacg acgacctgga caacctgctg gcccagatcg gcgaccagta
     1321 cgccgacctg tttctggccg ccaagaacct gtccgacgcc atcctgctga gcgacatcct
     1381 gagagtgaac accgagatca ccaaggcccc cctgagcgcc tctatgatca agagatacga
     1441 cgagcaccac caggacctga ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa
     1501 gtacaaagag attttcttcg accagagcaa gaacggctac gccggctaca ttgacggcgg
     1561 agccagccag gaagagttct acaagttcat caagcccatc ctggaaaaga tggacggcac
     1621 cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc ggaccttcga
     1681 caacggcagc atcccccacc agatccacct gggagagctg cacgccattc tgcggcggca
     1741 ggaagatttt tacccattcc tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt
     1801 ccgcatcccc tactacgtgg gccctctggc caggggaaac agcagattcg cctggatgac
     1861 cagaaagagc gaggaaacca tcaccccctg gaacttcgag gaagtggtgg acaagggcgc
     1921 ttccgcccag agcttcatcg agcggatgac caacttcgat aagaacctgc ccaacgagaa
     1981 ggtgctgccc aagcacagcc tgctgtacga gtacttcacc gtgtataacg agctgaccaa
     2041 agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa
     2101 ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga
     2161 ggactacttc aagaaaatcg agtgcttcga ctccgtggaa atctccggcg tggaagatcg
     2221 gttcaacgcc tccctgggca cataccacga tctgctgaaa attatcaagg acaaggactt
     2281 cctggacaat gaggaaaacg aggacattct ggaagatatc gtgctgaccc tgacactgtt
     2341 tgaggacaga gagatgatcg aggaacggct gaaaacctat gcccacctgt tcgacgacaa
     2401 agtgatgaag cagctgaagc ggcggagata caccggctgg ggcaggctga gccggaagct
     2461 gatcaacggc atccgggaca agcagtccgg caagacaatc ctggatttcc tgaagtccga
     2521 cggcttcgcc aacagaaact tcatgcagct gatccacgac gacagcctga cctttaaaga
     2581 ggacatccag aaagcccagg tgtccggcca gggcgatagc ctgcacgagc acattgccaa
     2641 tctggccggc agccccgcca ttaagaaggg catcctgcag acagtgaagg tggtggacga
     2701 gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga
     2761 gaaccagacc acccagaagg gacagaagaa cagccgcgag agaatgaagc ggatcgaaga
     2821 gggcatcaaa gagctgggca gccagatcct gaaagaacac cccgtggaaa acacccagct
     2881 gcagaacgag aagctgtacc tgtactacct gcagaatggg cgggatatgt acgtggacca
     2941 ggaactggac atcaaccggc tgtccgacta cgatgtggac catatcgtgc ctcagagctt
     3001 tctgaaggac gactccatcg acaacaaggt gctgaccaga agcgacaaga accggggcaa
     3061 gagcgacaac gtgccctccg aagaggtcgt gaagaagatg aagaactact ggcggcagct
     3121 gctgaacgcc aagctgatta cccagagaaa gttcgacaat ctgaccaagg ccgagagagg
     3181 cggcctgagc gaactggata aggccggctt catcaagaga cagctggtgg aaacccggca
     3241 gatcacaaag cacgtggcac agatcctgga ctcccggatg aacactaagt acgacgagaa
     3301 tgacaagctg atccgggaag tgaaagtgat caccctgaag tccaagctgg tgtccgattt
     3361 ccggaaggat ttccagtttt acaaagtgcg cgagatcaac aactaccacc acgcccacga
     3421 cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctggaaag
     3481 cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga
     3541 gcaggaaatc ggcaaggcta ccgccaagta cttcttctac agcaacatca tgaacttttt
     3601 caagaccgag attaccctgg ccaacggcga gatccggaag cggcctctga tcgagacaaa
     3661 cggcgaaacc ggggagatcg tgtgggataa gggccgggat tttgccaccg tgcggaaagt
     3721 gctgagcatg ccccaagtga atatcgtgaa aaagaccgag gtgcagacag gcggcttcag
     3781 caaagagtct atcctgccca agaggaacag cgataagctg atcgccagaa agaaggactg
     3841 ggaccctaag aagtacggcg gcttcgacag ccccaccgtg gcctattctg tgctggtggt
     3901 ggccaaagtg gaaaagggca agtccaagaa actgaagagt gtgaaagagc tgctggggat
     3961 caccatcatg gaaagaagca gcttcgagaa gaatcccatc gactttctgg aagccaaggg
     4021 ctacaaagaa gtgaaaaagg acctgatcat caagctgcct aagtactccc tgttcgagct
     4081 ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact
     4141 ggccctgccc tccaaatatg tgaacttcct gtacctggcc agccactatg agaagctgaa
     4201 gggctccccc gaggataatg agcagaaaca gctgtttgtg gaacagcaca agcactacct
     4261 ggacgagatc atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa
     4321 tctggacaaa gtgctgtccg cctacaacaa gcaccgggat aagcccatca gagagcaggc
     4381 cgagaatatc atccacctgt ttaccctgac caatctggga gcccctgccg ccttcaagta
     4441 ctttgacacc accatcgacc ggaagaggta caccagcacc aaagaggtgc tggacgccac
     4501 cctgatccac cagagcatca ccggcctgta cgagacacgg atcgacctgt ctcagctggg
     4561 aggcgacagc cccaagaaga agagaaaggt ggaggccagc taagaattct aactagagct
     4621 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     4681 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     4741 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     4801 agcaaggggg aggattggga agagaatagc aggcatgctg gggagcggcc gcaggaaccc
     4861 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     4921 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     4981 agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     5041 caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg
     5101 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt
     5161 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     5221 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     5281 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     5341 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact
     5401 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa
     5461 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     5521 ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     5581 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     5641 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     5701 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     5761 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     5821 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     5881 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     5941 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     6001 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     6061 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     6121 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     6181 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     6241 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     6301 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     6361 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     6421 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     6481 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     6541 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     6601 ataaatctgg agccggtgag cgtggaagcc gcggtatcat tgcagcactg gggccagatg
     6661 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     6721 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     6781 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     6841 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     6901 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     6961 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     7021 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     7081 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     7141 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     7201 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     7261 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     7321 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     7381 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     7441 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     7501 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     7561 tggccttttg ctggcctttt gctcacatgt gagggcctat ttcccatgat tccttcatat
     7621 ttgcatatac gatacaaggc tgttagagag ataattggaa ttaatttgac tgtaaacaca
     7681 aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt
     7741 ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat
     7801 ttcttggctt tatatatctt gtggaaagga cgaaacaccg gataggagtt tactatgacg
     7861 ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg
     7921 gcaccgagtc ggtgcttttt tgttttagag ctagaaatag caagttaaaa taaggctagt
     7981 ccgtttttag cgcgtgcgcc aattctgcag acaaatggct ctagaagctt ggataggagt
     8041 ttactatgac gggatctgct agcggtagcg gctccagtag atctggggtg agcaagggcg
     8101 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     8161 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     8221 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     8281 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     8341 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     8401 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     8461 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     8521 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     8581 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     8641 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     8701 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     8761 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaggcctag gataggagtt
     8821 tactatgacg ggctcgagga tcctaggacg cgtcctagga cgcgtccaga ggtacccgtt
     8881 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     8941 tcaatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     9001 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     9061 aatgacggta aatggcccgc ctggcattgt gcccagtaca tgaccttatg ggactttcct
     9121 acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg
     9181 ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt
     9241 ttttaattat tttgtgcagc gatgggggcg ggggg
//
