LOCUS       sequence-269653-        9299 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  G3BP1 gRNA (#1) inserted into the pSpCas9(BB)-2A-GFP plasmid 
            (GTAGTCCCCTGCTGGTCGGGC).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9299)
  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 9299)
  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-G3BP1(1)
FEATURES             Location/Qualifiers
     source          1..9299
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      233..242
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             242..307
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             314..334
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             359..4459
                     /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             359..4459
                     /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             4460..4507
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4523..4576
                     /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(4577..4579,4580..5290)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(4577..4579,4580..5290)
                     /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(4880..4899)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5227..5248
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5318..5335)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5324..5531
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5540..5680
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5540..5669
                     /label=AAV2 ITR
     rep_origin      5755..6210
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5842..5861)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6052..6073
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6227..6246)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6346..6368
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6406..6424)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6492..6596
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6597..6665,6666..7457)
                     /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(6815..6834)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7628..8216
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8117..8136
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8278..8526
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8278..8298
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8449..8468
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8548..8623
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8720..9005
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     intron          join(9293..9299,1..221)
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
ORIGIN
        1 ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc
       61 tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct cctccgggct
      121 gtaattagct gagcaagagg taagggttta agggatggtt ggttggtggg gtattaatgt
      181 ttaattacct ggagcacctg cctgaaatca ctttttttca ggttggaccg gtgccaccat
      241 ggactataag gaccacgacg gagactacaa ggatcatgat attgattaca aagacgatga
      301 cgataagatg gccccaaaga agaagcggaa ggtcggtatc cacggagtcc cagcagccga
      361 caagaagtac agcatcggcc tggacatcgg caccaactct gtgggctggg ccgtgatcac
      421 cgacgagtac aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg accggcacag
      481 catcaagaag aacctgatcg gagccctgct gttcgacagc ggcgaaacag ccgaggccac
      541 ccggctgaag agaaccgcca gaagaagata caccagacgg aagaaccgga tctgctatct
      601 gcaagagatc ttcagcaacg agatggccaa ggtggacgac agcttcttcc acagactgga
      661 agagtccttc ctggtggaag aggataagaa gcacgagcgg caccccatct tcggcaacat
      721 cgtggacgag gtggcctacc acgagaagta ccccaccatc taccacctga gaaagaaact
      781 ggtggacagc accgacaagg ccgacctgcg gctgatctat ctggccctgg cccacatgat
      841 caagttccgg ggccacttcc tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga
      901 caagctgttc atccagctgg tgcagaccta caaccagctg ttcgaggaaa accccatcaa
      961 cgccagcggc gtggacgcca aggccatcct gtctgccaga ctgagcaaga gcagacggct
     1021 ggaaaatctg atcgcccagc tgcccggcga gaagaagaat ggcctgttcg gaaacctgat
     1081 tgccctgagc ctgggcctga cccccaactt caagagcaac ttcgacctgg ccgaggatgc
     1141 caaactgcag ctgagcaagg acacctacga cgacgacctg gacaacctgc tggcccagat
     1201 cggcgaccag tacgccgacc tgtttctggc cgccaagaac ctgtccgacg ccatcctgct
     1261 gagcgacatc ctgagagtga acaccgagat caccaaggcc cccctgagcg cctctatgat
     1321 caagagatac gacgagcacc accaggacct gaccctgctg aaagctctcg tgcggcagca
     1381 gctgcctgag aagtacaaag agattttctt cgaccagagc aagaacggct acgccggcta
     1441 cattgacggc ggagccagcc aggaagagtt ctacaagttc atcaagccca tcctggaaaa
     1501 gatggacggc accgaggaac tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca
     1561 gcggaccttc gacaacggca gcatccccca ccagatccac ctgggagagc tgcacgccat
     1621 tctgcggcgg caggaagatt tttacccatt cctgaaggac aaccgggaaa agatcgagaa
     1681 gatcctgacc ttccgcatcc cctactacgt gggccctctg gccaggggaa acagcagatt
     1741 cgcctggatg accagaaaga gcgaggaaac catcaccccc tggaacttcg aggaagtggt
     1801 ggacaagggc gcttccgccc agagcttcat cgagcggatg accaacttcg ataagaacct
     1861 gcccaacgag aaggtgctgc ccaagcacag cctgctgtac gagtacttca ccgtgtataa
     1921 cgagctgacc aaagtgaaat acgtgaccga gggaatgaga aagcccgcct tcctgagcgg
     1981 cgagcagaaa aaggccatcg tggacctgct gttcaagacc aaccggaaag tgaccgtgaa
     2041 gcagctgaaa gaggactact tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg
     2101 cgtggaagat cggttcaacg cctccctggg cacataccac gatctgctga aaattatcaa
     2161 ggacaaggac ttcctggaca atgaggaaaa cgaggacatt ctggaagata tcgtgctgac
     2221 cctgacactg tttgaggaca gagagatgat cgaggaacgg ctgaaaacct atgcccacct
     2281 gttcgacgac aaagtgatga agcagctgaa gcggcggaga tacaccggct ggggcaggct
     2341 gagccggaag ctgatcaacg gcatccggga caagcagtcc ggcaagacaa tcctggattt
     2401 cctgaagtcc gacggcttcg ccaacagaaa cttcatgcag ctgatccacg acgacagcct
     2461 gacctttaaa gaggacatcc agaaagccca ggtgtccggc cagggcgata gcctgcacga
     2521 gcacattgcc aatctggccg gcagccccgc cattaagaag ggcatcctgc agacagtgaa
     2581 ggtggtggac gagctcgtga aagtgatggg ccggcacaag cccgagaaca tcgtgatcga
     2641 aatggccaga gagaaccaga ccacccagaa gggacagaag aacagccgcg agagaatgaa
     2701 gcggatcgaa gagggcatca aagagctggg cagccagatc ctgaaagaac accccgtgga
     2761 aaacacccag ctgcagaacg agaagctgta cctgtactac ctgcagaatg ggcgggatat
     2821 gtacgtggac caggaactgg acatcaaccg gctgtccgac tacgatgtgg accatatcgt
     2881 gcctcagagc tttctgaagg acgactccat cgacaacaag gtgctgacca gaagcgacaa
     2941 gaaccggggc aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta
     3001 ctggcggcag ctgctgaacg ccaagctgat tacccagaga aagttcgaca atctgaccaa
     3061 ggccgagaga ggcggcctga gcgaactgga taaggccggc ttcatcaaga gacagctggt
     3121 ggaaacccgg cagatcacaa agcacgtggc acagatcctg gactcccgga tgaacactaa
     3181 gtacgacgag aatgacaagc tgatccggga agtgaaagtg atcaccctga agtccaagct
     3241 ggtgtccgat ttccggaagg atttccagtt ttacaaagtg cgcgagatca acaactacca
     3301 ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc
     3361 taagctggaa agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat
     3421 cgccaagagc gagcaggaaa tcggcaaggc taccgccaag tacttcttct acagcaacat
     3481 catgaacttt ttcaagaccg agattaccct ggccaacggc gagatccgga agcggcctct
     3541 gatcgagaca aacggcgaaa ccggggagat cgtgtgggat aagggccggg attttgccac
     3601 cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac
     3661 aggcggcttc agcaaagagt ctatcctgcc caagaggaac agcgataagc tgatcgccag
     3721 aaagaaggac tgggacccta agaagtacgg cggcttcgac agccccaccg tggcctattc
     3781 tgtgctggtg gtggccaaag tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga
     3841 gctgctgggg atcaccatca tggaaagaag cagcttcgag aagaatccca tcgactttct
     3901 ggaagccaag ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc
     3961 cctgttcgag ctggaaaacg gccggaagag aatgctggcc tctgccggcg aactgcagaa
     4021 gggaaacgaa ctggccctgc cctccaaata tgtgaacttc ctgtacctgg ccagccacta
     4081 tgagaagctg aagggctccc ccgaggataa tgagcagaaa cagctgtttg tggaacagca
     4141 caagcactac ctggacgaga tcatcgagca gatcagcgag ttctccaaga gagtgatcct
     4201 ggccgacgct aatctggaca aagtgctgtc cgcctacaac aagcaccggg ataagcccat
     4261 cagagagcag gccgagaata tcatccacct gtttaccctg accaatctgg gagcccctgc
     4321 cgccttcaag tactttgaca ccaccatcga ccggaagagg tacaccagca ccaaagaggt
     4381 gctggacgcc accctgatcc accagagcat caccggcctg tacgagacac ggatcgacct
     4441 gtctcagctg ggaggcgaca aaaggccggc ggccacgaaa aaggccggcc aggcaaaaaa
     4501 gaaaaaggaa ttcggcagtg gagagggcag aggaagtctg ctaacatgcg gtgacgtcga
     4561 ggagaatcct ggcccagtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     4621 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     4681 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     4741 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     4801 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     4861 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     4921 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     4981 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     5041 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     5101 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     5161 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     5221 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     5281 gctgtacaag gaattctaac tagagctcgc tgatcagcct cgactgtgcc ttctagttgc
     5341 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     5401 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     5461 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga gaatagcagg
     5521 catgctgggg agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     5581 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     5641 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt
     5701 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     5761 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     5821 ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     5881 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     5941 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     6001 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     6061 tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga
     6121 ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     6181 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     6241 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     6301 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     6361 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     6421 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     6481 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     6541 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     6601 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     6661 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     6721 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     6781 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     6841 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     6901 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     6961 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     7021 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     7081 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     7141 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     7201 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     7261 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt ggaagccgcg
     7321 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     7381 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     7441 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     7501 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     7561 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     7621 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     7681 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     7741 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     7801 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     7861 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     7921 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     7981 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     8041 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     8101 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     8161 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     8221 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtgag
     8281 ggcctatttc ccatgattcc ttcatatttg catatacgat acaaggctgt tagagagata
     8341 attggaatta atttgactgt aaacacaaag atattagtac aaaatacgtg acgtagaaag
     8401 taataatttc ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga ctatcatatg
     8461 cttaccgtaa cttgaaagta tttcgatttc ttggctttat atatcttgtg gaaaggacga
     8521 aacaccgtag tcccctgctg gtcgggcgtt ttagagctag aaatagcaag ttaaaataag
     8581 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg ttttagagct
     8641 agaaatagca agttaaaata aggctagtcc gtttttagcg cgtgcgccaa ttctgcagac
     8701 aaatggctct agaggtaccc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     8761 ccaacgaccc ccgcccattg acgtcaatag taacgccaat agggactttc cattgacgtc
     8821 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     8881 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tgtgcccagt
     8941 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     9001 ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac
     9061 ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg
     9121 gggggggggg ngggsssmgg gggngggggg gggggggggg ggsgrggggs ggggcggggc
     9181 gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat
     9241 ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcg
//
