LOCUS       sequence-292309-        9290 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  G3BP2 gRNA (#1) inserted into the pSpCas9(BB)-2A-GFP plasmid 
            (TCATACTAAAATTCGTCATG).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9290)
  AUTHORS   Fischer JW, Busa VF, Shao Y, Leung AKL
  TITLE     Structure-Mediated RNA Decay by UPF1 and G3BP1.
  JOURNAL   Mol Cell. 2020 Apr 2;78(1):70-84.e6. doi: 
            10.1016/j.molcel.2020.01.021. Epub 2020 Feb 3.
  PUBMED    32017897
REFERENCE   2  (bases 1 to 9290)
  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"; doi: 
            "10.1016/j.molcel.2020.01.021"; journalName: "Mol Cell"; date: 
            "2020-04-2- 2"; volume: "78"; issue: "1"; pages: "70-84.e6"
COMMENT     Sequence Label: pSpCas9(BB)-2A-GFP-G3BP2(1)
FEATURES             Location/Qualifiers
     source          1..9290
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..353
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        355..632
                     /label=chicken beta-actin promoter
     intron          633..860
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      872..881
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             881..946
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             953..973
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             998..5098
                     /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             998..5098
                     /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             5099..5146
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5162..5215
                     /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             join(5216..5218,5219..5929)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(5216..5218,5219..5929)
                     /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(5519..5538)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5866..5887
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5957..5974)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5963..6170
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6179..6319
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6179..6308
                     /label=AAV2 ITR
     rep_origin      6394..6849
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6481..6500)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6691..6712
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6866..6885)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6985..7007
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7045..7063)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7131..7235
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7236..7304,7305..8096)
                     /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(7454..7473)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8267..8855
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8756..8775
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8917..9165
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8917..8937
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     9088..9107
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        9186..9261
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 ttaaaataag gctagtccgt ttttagcgcg tgcgccaatt ctgcagacaa atggctctag
       61 aggtacccgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
      181 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
      241 ctattgacgt caatgacggt aaatggcccg cctggcattg tgcccagtac atgaccttat
      301 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtcgagg
      361 tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc ccaattttgt
      421 atttatttat tttttaatta ttttgtgcag cgatgggggc gggggggggg ggggggcgcg
      481 cgccrggsgg ggsggggsgg ggggsggggs ggggsggggc gaggcggaga ggtgcggcgg
      541 cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc
      601 ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg
      661 tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc
      721 cacaggtgag cgggcgggac ggcccttctc ctccgggctg taattagctg agcaagaggt
      781 aagggtttaa gggatggttg gttggtgggg tattaatgtt taattacctg gagcacctgc
      841 ctgaaatcac tttttttcag gttggaccgg tgccaccatg gactataagg accacgacgg
      901 agactacaag gatcatgata ttgattacaa agacgatgac gataagatgg ccccaaagaa
      961 gaagcggaag gtcggtatcc acggagtccc agcagccgac aagaagtaca gcatcggcct
     1021 ggacatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag
     1081 caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg
     1141 agccctgctg ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga gaaccgccag
     1201 aagaagatac accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga
     1261 gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga
     1321 ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca
     1381 cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc
     1441 cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct
     1501 gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt
     1561 gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa
     1621 ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga tcgcccagct
     1681 gcccggcgag aagaagaatg gcctgttcgg aaacctgatt gccctgagcc tgggcctgac
     1741 ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc tgagcaagga
     1801 cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt acgccgacct
     1861 gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc tgagagtgaa
     1921 caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg acgagcacca
     1981 ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga agtacaaaga
     2041 gattttcttc gaccagagca agaacggcta cgccggctac attgacggcg gagccagcca
     2101 ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca ccgaggaact
     2161 gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg acaacggcag
     2221 catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc aggaagattt
     2281 ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct tccgcatccc
     2341 ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga ccagaaagag
     2401 cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg cttccgccca
     2461 gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc
     2521 caagcacagc ctgctgtacg agtacttcac cgtgtataac gagctgacca aagtgaaata
     2581 cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa aggccatcgt
     2641 ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag aggactactt
     2701 caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc ggttcaacgc
     2761 ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact tcctggacaa
     2821 tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag
     2881 agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa
     2941 gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg
     3001 catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc
     3061 caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca
     3121 gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca atctggccgg
     3181 cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg agctcgtgaa
     3241 agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag agaaccagac
     3301 cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag agggcatcaa
     3361 agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc tgcagaacga
     3421 gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc aggaactgga
     3481 catcaaccgg ctgtccgact acgatgtgga ccatatcgtg cctcagagct ttctgaagga
     3541 cgactccatc gacaacaagg tgctgaccag aagcgacaag aaccggggca agagcgacaa
     3601 cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcggcagc tgctgaacgc
     3661 caagctgatt acccagagaa agttcgacaa tctgaccaag gccgagagag gcggcctgag
     3721 cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc agatcacaaa
     3781 gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga atgacaagct
     3841 gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccggaagga
     3901 tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg acgcctacct
     3961 gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt
     4021 gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg agcaggaaat
     4081 cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt tcaagaccga
     4141 gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa acggcgaaac
     4201 cggggagatc gtgtgggata agggccggga ttttgccacc gtgcggaaag tgctgagcat
     4261 gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc
     4321 tatcctgccc aagaggaaca gcgataagct gatcgccaga aagaaggact gggaccctaa
     4381 gaagtacggc ggcttcgaca gccccaccgt ggcctattct gtgctggtgg tggccaaagt
     4441 ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctgctgggga tcaccatcat
     4501 ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg gctacaaaga
     4561 agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg
     4621 ccggaagaga atgctggcct ctgccggcga actgcagaag ggaaacgaac tggccctgcc
     4681 ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga agggctcccc
     4741 cgaggataat gagcagaaac agctgtttgt ggaacagcac aagcactacc tggacgagat
     4801 catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta atctggacaa
     4861 agtgctgtcc gcctacaaca agcaccggga taagcccatc agagagcagg ccgagaatat
     4921 catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt actttgacac
     4981 caccatcgac cggaagaggt acaccagcac caaagaggtg ctggacgcca ccctgatcca
     5041 ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg gaggcgacaa
     5101 aaggccggcg gccacgaaaa aggccggcca ggcaaaaaag aaaaaggaat tcggcagtgg
     5161 agagggcaga ggaagtctgc taacatgcgg tgacgtcgag gagaatcctg gcccagtgag
     5221 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     5281 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     5341 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     5401 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     5461 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     5521 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     5581 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     5641 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     5701 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     5761 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     5821 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     5881 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg aattctaact
     5941 agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     6001 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     6061 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     6121 caggacagca agggggagga ttgggaagag aatagcaggc atgctgggga gcggccgcag
     6181 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     6241 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
     6301 gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     6361 atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg
     6421 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg
     6481 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     6541 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     6601 aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     6661 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     6721 tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt
     6781 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     6841 ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     6901 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     6961 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     7021 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     7081 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     7141 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     7201 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     7261 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     7321 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     7381 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     7441 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     7501 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     7561 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     7621 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     7681 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     7741 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     7801 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     7861 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     7921 ttgctgataa atctggagcc ggtgagcgtg gaagccgcgg tatcattgca gcactggggc
     7981 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     8041 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     8101 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     8161 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     8221 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     8281 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     8341 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     8401 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     8461 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     8521 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     8581 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     8641 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     8701 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     8761 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     8821 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     8881 ggttcctggc cttttgctgg ccttttgctc acatgtgagg gcctatttcc catgattcct
     8941 tcatatttgc atatacgata caaggctgtt agagagataa ttggaattaa tttgactgta
     9001 aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt
     9061 gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat
     9121 ttcgatttct tggctttata tatcttgtgg aaaggacgaa acacctcata ctaaaattcg
     9181 tcatggtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa
     9241 aaagtggcac cgagtcggtg cttttttgtt ttagagctag aaatagcaag
//
