LOCUS       sequence-385015-        9176 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cas9 + gRNA expression to edit G3BP1 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9176)
  AUTHORS   Sidhaye J, Trepte P, Sepke N, Novatchkova M, Schutzbier M, 
            Durnberger G, Mechtler K, Knoblich JA
  TITLE     Integrated transcriptome and proteome analysis reveals 
            posttranscriptional regulation of ribosomal genes in human brain 
            organoids.
  JOURNAL   Elife. 2023 Mar 29;12:e85135. doi: 10.7554/eLife.85135.
  PUBMED    36989136
REFERENCE   2  (bases 1 to 9176)
  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: "Elife."; 
            date: "2023-03-29"; volume: "12:e85135. doi"; pages: " 
            10.7554/eLife.85135"
COMMENT     Sequence Label: pSpCas9(BB)-2A-Puro G3BP1-sgRNA
FEATURES             Location/Qualifiers
     source          1..9176
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      769..1357
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1258..1277
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1419..1667
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1419..1439
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1590..1609
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        1689..1764
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        1861..2146
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     intron          2425..2652
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      2664..2673
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2673..2738
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2745..2765
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2790..6890
                     /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             2790..6890
                     /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             6891..6938
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6954..7007
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             7008..7607
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(7008..7027)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7504..7524
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(7635..7652)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7641..7848
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   7857..7997
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   7857..7986
                     /label=AAV2 ITR
     rep_origin      8072..8527
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8159..8178)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8369..8390
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8544..8563)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8663..8685
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8723..8741)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8809..8913
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8914..8982,8983..9176,1..598)
                     /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(9132..9151)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
       61 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
      121 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
      181 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
      241 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
      301 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
      361 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
      421 tattgctgat aaatctggag ccggtgagcg tggaagccgc ggtatcattg cagcactggg
      481 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
      541 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
      601 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
      661 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
      721 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
      781 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
      841 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
      901 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
      961 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     1021 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     1081 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     1141 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     1201 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     1261 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     1321 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     1381 acggttcctg gccttttgct ggccttttgc tcacatgtga gggcctattt cccatgattc
     1441 cttcatattt gcatatacga tacaaggctg ttagagagat aattggaatt aatttgactg
     1501 taaacacaaa gatattagta caaaatacgt gacgtagaaa gtaataattt cttgggtagt
     1561 ttgcagtttt aaaattatgt tttaaaatgg actatcatat gcttaccgta acttgaaagt
     1621 atttcgattt cttggcttta tatatcttgt ggaaaggacg aaacaccgtc catgaagatt
     1681 cactgccggt tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt
     1741 gaaaaagtgg caccgagtcg gtgctttttt gttttagagc tagaaatagc aagttaaaat
     1801 aaggctagtc cgtttttagc gcgtgcgcca attctgcaga caaatggctc tagaggtacc
     1861 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     1921 gacgtcaata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     1981 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     2041 cgtcaatgac ggtaaatggc ccgcctggca ttgtgcccag tacatgacct tatgggactt
     2101 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc
     2161 cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt
     2221 tattttttaa ttattttgtg cagcgatggg ggcggggggg ggggggggcg cgcgccrggg
     2281 ggggsggggs ggggsgrggg gsggggsggg gsgrggsggr grggkgcggc ggcagccaat
     2341 cagagcggcg cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg gcggccctat
     2401 aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc
     2461 tccgccgccg cctcgcgccg cccgccccgg ctctgactga ccgcgttact cccacaggtg
     2521 agcgggcggg acggcccttc tcctccgggc tgtaattagc tgagcaagag gtaagggttt
     2581 aagggatggt tggttggtgg ggtattaatg tttaattacc tggagcacct gcctgaaatc
     2641 actttttttc aggttggacc ggtgccacca tggactataa ggaccacgac ggagactaca
     2701 aggatcatga tattgattac aaagacgatg acgataagat ggccccaaag aagaagcgga
     2761 aggtcggtat ccacggagtc ccagcagccg acaagaagta cagcatcggc ctggacatcg
     2821 gcaccaactc tgtgggctgg gccgtgatca ccgacgagta caaggtgccc agcaagaaat
     2881 tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa gaacctgatc ggagccctgc
     2941 tgttcgacag cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc agaagaagat
     3001 acaccagacg gaagaaccgg atctgctatc tgcaagagat cttcagcaac gagatggcca
     3061 aggtggacga cagcttcttc cacagactgg aagagtcctt cctggtggaa gaggataaga
     3121 agcacgagcg gcaccccatc ttcggcaaca tcgtggacga ggtggcctac cacgagaagt
     3181 accccaccat ctaccacctg agaaagaaac tggtggacag caccgacaag gccgacctgc
     3241 ggctgatcta tctggccctg gcccacatga tcaagttccg gggccacttc ctgatcgagg
     3301 gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt catccagctg gtgcagacct
     3361 acaaccagct gttcgaggaa aaccccatca acgccagcgg cgtggacgcc aaggccatcc
     3421 tgtctgccag actgagcaag agcagacggc tggaaaatct gatcgcccag ctgcccggcg
     3481 agaagaagaa tggcctgttc ggaaacctga ttgccctgag cctgggcctg acccccaact
     3541 tcaagagcaa cttcgacctg gccgaggatg ccaaactgca gctgagcaag gacacctacg
     3601 acgacgacct ggacaacctg ctggcccaga tcggcgacca gtacgccgac ctgtttctgg
     3661 ccgccaagaa cctgtccgac gccatcctgc tgagcgacat cctgagagtg aacaccgaga
     3721 tcaccaaggc ccccctgagc gcctctatga tcaagagata cgacgagcac caccaggacc
     3781 tgaccctgct gaaagctctc gtgcggcagc agctgcctga gaagtacaaa gagattttct
     3841 tcgaccagag caagaacggc tacgccggct acattgacgg cggagccagc caggaagagt
     3901 tctacaagtt catcaagccc atcctggaaa agatggacgg caccgaggaa ctgctcgtga
     3961 agctgaacag agaggacctg ctgcggaagc agcggacctt cgacaacggc agcatccccc
     4021 accagatcca cctgggagag ctgcacgcca ttctgcggcg gcaggaagat ttttacccat
     4081 tcctgaagga caaccgggaa aagatcgaga agatcctgac cttccgcatc ccctactacg
     4141 tgggccctct ggccagggga aacagcagat tcgcctggat gaccagaaag agcgaggaaa
     4201 ccatcacccc ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc cagagcttca
     4261 tcgagcggat gaccaacttc gataagaacc tgcccaacga gaaggtgctg cccaagcaca
     4321 gcctgctgta cgagtacttc accgtgtata acgagctgac caaagtgaaa tacgtgaccg
     4381 agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc
     4441 tgttcaagac caaccggaaa gtgaccgtga agcagctgaa agaggactac ttcaagaaaa
     4501 tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga tcggttcaac gcctccctgg
     4561 gcacatacca cgatctgctg aaaattatca aggacaagga cttcctggac aatgaggaaa
     4621 acgaggacat tctggaagat atcgtgctga ccctgacact gtttgaggac agagagatga
     4681 tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga caaagtgatg aagcagctga
     4741 agcggcggag atacaccggc tggggcaggc tgagccggaa gctgatcaac ggcatccggg
     4801 acaagcagtc cggcaagaca atcctggatt tcctgaagtc cgacggcttc gccaacagaa
     4861 acttcatgca gctgatccac gacgacagcc tgacctttaa agaggacatc cagaaagccc
     4921 aggtgtccgg ccagggcgat agcctgcacg agcacattgc caatctggcc ggcagccccg
     4981 ccattaagaa gggcatcctg cagacagtga aggtggtgga cgagctcgtg aaagtgatgg
     5041 gccggcacaa gcccgagaac atcgtgatcg aaatggccag agagaaccag accacccaga
     5101 agggacagaa gaacagccgc gagagaatga agcggatcga agagggcatc aaagagctgg
     5161 gcagccagat cctgaaagaa caccccgtgg aaaacaccca gctgcagaac gagaagctgt
     5221 acctgtacta cctgcagaat gggcgggata tgtacgtgga ccaggaactg gacatcaacc
     5281 ggctgtccga ctacgatgtg gaccatatcg tgcctcagag ctttctgaag gacgactcca
     5341 tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg caagagcgac aacgtgccct
     5401 ccgaagaggt cgtgaagaag atgaagaact actggcggca gctgctgaac gccaagctga
     5461 ttacccagag aaagttcgac aatctgacca aggccgagag aggcggcctg agcgaactgg
     5521 ataaggccgg cttcatcaag agacagctgg tggaaacccg gcagatcaca aagcacgtgg
     5581 cacagatcct ggactcccgg atgaacacta agtacgacga gaatgacaag ctgatccggg
     5641 aagtgaaagt gatcaccctg aagtccaagc tggtgtccga tttccggaag gatttccagt
     5701 tttacaaagt gcgcgagatc aacaactacc accacgccca cgacgcctac ctgaacgccg
     5761 tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga aagcgagttc gtgtacggcg
     5821 actacaaggt gtacgacgtg cggaagatga tcgccaagag cgagcaggaa atcggcaagg
     5881 ctaccgccaa gtacttcttc tacagcaaca tcatgaactt tttcaagacc gagattaccc
     5941 tggccaacgg cgagatccgg aagcggcctc tgatcgagac aaacggcgaa accggggaga
     6001 tcgtgtggga taagggccgg gattttgcca ccgtgcggaa agtgctgagc atgccccaag
     6061 tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt cagcaaagag tctatcctgc
     6121 ccaagaggaa cagcgataag ctgatcgcca gaaagaagga ctgggaccct aagaagtacg
     6181 gcggcttcga cagccccacc gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg
     6241 gcaagtccaa gaaactgaag agtgtgaaag agctgctggg gatcaccatc atggaaagaa
     6301 gcagcttcga gaagaatccc atcgactttc tggaagccaa gggctacaaa gaagtgaaaa
     6361 aggacctgat catcaagctg cctaagtact ccctgttcga gctggaaaac ggccggaaga
     6421 gaatgctggc ctctgccggc gaactgcaga agggaaacga actggccctg ccctccaaat
     6481 atgtgaactt cctgtacctg gccagccact atgagaagct gaagggctcc cccgaggata
     6541 atgagcagaa acagctgttt gtggaacagc acaagcacta cctggacgag atcatcgagc
     6601 agatcagcga gttctccaag agagtgatcc tggccgacgc taatctggac aaagtgctgt
     6661 ccgcctacaa caagcaccgg gataagccca tcagagagca ggccgagaat atcatccacc
     6721 tgtttaccct gaccaatctg ggagcccctg ccgccttcaa gtactttgac accaccatcg
     6781 accggaagag gtacaccagc accaaagagg tgctggacgc caccctgatc caccagagca
     6841 tcaccggcct gtacgagaca cggatcgacc tgtctcagct gggaggcgac aaaaggccgg
     6901 cggccacgaa aaaggccggc caggcaaaaa agaaaaagga attcggcagt ggagagggca
     6961 gaggaagtct gctaacatgc ggtgacgtcg aggagaatcc tggcccaatg accgagtaca
     7021 agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg
     7081 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc
     7141 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
     7201 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg
     7261 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg
     7321 cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
     7381 tggccaccgt cggagtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
     7441 tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc
     7501 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc
     7561 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgagaa ttctaactag
     7621 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     7681 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     7741 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     7801 ggacagcaag ggggaggatt gggaagagaa tagcaggcat gctggggagc ggccgcagga
     7861 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     7921 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     7981 gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     8041 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg
     8101 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct
     8161 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     8221 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     8281 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     8341 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     8401 aactctatct cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg
     8461 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     8521 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     8581 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     8641 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     8701 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     8761 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     8821 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     8881 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     8941 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     9001 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     9061 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     9121 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagca
//
