LOCUS       sequence-269656-        9290 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  G3BP1 gRNA (#2) inserted into the pSpCas9(BB)-2A-GFP plasmid 
            (GTATTACACACTGCTGAACC).
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-G3BP1(2)
FEATURES             Location/Qualifiers
     source          1..9290
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        65..140
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        237..522
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        524..801
                     /label=chicken beta-actin promoter
     intron          802..1029
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1041..1050
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1050..1115
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1122..1142
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1167..5267
                     /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             1167..5267
                     /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             5268..5315
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5331..5384
                     /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(5385..5387,5388..6098)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(5385..5387,5388..6098)
                     /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(5688..5707)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6035..6056
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6126..6143)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6132..6339
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6348..6488
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6348..6477
                     /label=AAV2 ITR
     rep_origin      6563..7018
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6650..6669)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6860..6881
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7035..7054)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7154..7176
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7214..7232)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7300..7404
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7405..7473,7474..8265)
                     /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(7623..7642)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8436..9024
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8925..8944
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(9086..9290,1..44)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     9086..9106
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     9257..9276
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 tcgatttctt ggctttatat atcttgtgga aaggacgaaa caccgtatta cacactgctg
       61 aaccgtttta gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa
      121 aagtggcacc gagtcggtgc ttttttgttt tagagctaga aatagcaagt taaaataagg
      181 ctagtccgtt tttagcgcgt gcgccaattc tgcagacaaa tggctctaga ggtacccgtt
      241 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
      301 tcaatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      361 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      421 aatgacggta aatggcccgc ctggcattgt gcccagtaca tgaccttatg ggactttcct
      481 acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg
      541 ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt
      601 ttttaattat tttgtgcagc gatgggggcg gggggggggg gggggcgcgc gccrgggggg
      661 gggggggggg gsgggggggg gggsggggsg rggcggagag gkgcggcggc agccaatcag
      721 agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg gccctataaa
      781 aagcgaagcg cgcggcgggc gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc
      841 gccgccgcct cgcgccgccc gccccggctc tgactgaccg cgttactccc acaggtgagc
      901 gggcgggacg gcccttctcc tccgggctgt aattagctga gcaagaggta agggtttaag
      961 ggatggttgg ttggtggggt attaatgttt aattacctgg agcacctgcc tgaaatcact
     1021 ttttttcagg ttggaccggt gccaccatgg actataagga ccacgacgga gactacaagg
     1081 atcatgatat tgattacaaa gacgatgacg ataagatggc cccaaagaag aagcggaagg
     1141 tcggtatcca cggagtccca gcagccgaca agaagtacag catcggcctg gacatcggca
     1201 ccaactctgt gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc aagaaattca
     1261 aggtgctggg caacaccgac cggcacagca tcaagaagaa cctgatcgga gccctgctgt
     1321 tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag aaccgccaga agaagataca
     1381 ccagacggaa gaaccggatc tgctatctgc aagagatctt cagcaacgag atggccaagg
     1441 tggacgacag cttcttccac agactggaag agtccttcct ggtggaagag gataagaagc
     1501 acgagcggca ccccatcttc ggcaacatcg tggacgaggt ggcctaccac gagaagtacc
     1561 ccaccatcta ccacctgaga aagaaactgg tggacagcac cgacaaggcc gacctgcggc
     1621 tgatctatct ggccctggcc cacatgatca agttccgggg ccacttcctg atcgagggcg
     1681 acctgaaccc cgacaacagc gacgtggaca agctgttcat ccagctggtg cagacctaca
     1741 accagctgtt cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag gccatcctgt
     1801 ctgccagact gagcaagagc agacggctgg aaaatctgat cgcccagctg cccggcgaga
     1861 agaagaatgg cctgttcgga aacctgattg ccctgagcct gggcctgacc cccaacttca
     1921 agagcaactt cgacctggcc gaggatgcca aactgcagct gagcaaggac acctacgacg
     1981 acgacctgga caacctgctg gcccagatcg gcgaccagta cgccgacctg tttctggccg
     2041 ccaagaacct gtccgacgcc atcctgctga gcgacatcct gagagtgaac accgagatca
     2101 ccaaggcccc cctgagcgcc tctatgatca agagatacga cgagcaccac caggacctga
     2161 ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa gtacaaagag attttcttcg
     2221 accagagcaa gaacggctac gccggctaca ttgacggcgg agccagccag gaagagttct
     2281 acaagttcat caagcccatc ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc
     2341 tgaacagaga ggacctgctg cggaagcagc ggaccttcga caacggcagc atcccccacc
     2401 agatccacct gggagagctg cacgccattc tgcggcggca ggaagatttt tacccattcc
     2461 tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc tactacgtgg
     2521 gccctctggc caggggaaac agcagattcg cctggatgac cagaaagagc gaggaaacca
     2581 tcaccccctg gaacttcgag gaagtggtgg acaagggcgc ttccgcccag agcttcatcg
     2641 agcggatgac caacttcgat aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc
     2701 tgctgtacga gtacttcacc gtgtataacg agctgaccaa agtgaaatac gtgaccgagg
     2761 gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa ggccatcgtg gacctgctgt
     2821 tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga ggactacttc aagaaaatcg
     2881 agtgcttcga ctccgtggaa atctccggcg tggaagatcg gttcaacgcc tccctgggca
     2941 cataccacga tctgctgaaa attatcaagg acaaggactt cctggacaat gaggaaaacg
     3001 aggacattct ggaagatatc gtgctgaccc tgacactgtt tgaggacaga gagatgatcg
     3061 aggaacggct gaaaacctat gcccacctgt tcgacgacaa agtgatgaag cagctgaagc
     3121 ggcggagata caccggctgg ggcaggctga gccggaagct gatcaacggc atccgggaca
     3181 agcagtccgg caagacaatc ctggatttcc tgaagtccga cggcttcgcc aacagaaact
     3241 tcatgcagct gatccacgac gacagcctga cctttaaaga ggacatccag aaagcccagg
     3301 tgtccggcca gggcgatagc ctgcacgagc acattgccaa tctggccggc agccccgcca
     3361 ttaagaaggg catcctgcag acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc
     3421 ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga gaaccagacc acccagaagg
     3481 gacagaagaa cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa gagctgggca
     3541 gccagatcct gaaagaacac cccgtggaaa acacccagct gcagaacgag aagctgtacc
     3601 tgtactacct gcagaatggg cgggatatgt acgtggacca ggaactggac atcaaccggc
     3661 tgtccgacta cgatgtggac catatcgtgc ctcagagctt tctgaaggac gactccatcg
     3721 acaacaaggt gctgaccaga agcgacaaga accggggcaa gagcgacaac gtgccctccg
     3781 aagaggtcgt gaagaagatg aagaactact ggcggcagct gctgaacgcc aagctgatta
     3841 cccagagaaa gttcgacaat ctgaccaagg ccgagagagg cggcctgagc gaactggata
     3901 aggccggctt catcaagaga cagctggtgg aaacccggca gatcacaaag cacgtggcac
     3961 agatcctgga ctcccggatg aacactaagt acgacgagaa tgacaagctg atccgggaag
     4021 tgaaagtgat caccctgaag tccaagctgg tgtccgattt ccggaaggat ttccagtttt
     4081 acaaagtgcg cgagatcaac aactaccacc acgcccacga cgcctacctg aacgccgtcg
     4141 tgggaaccgc cctgatcaaa aagtacccta agctggaaag cgagttcgtg tacggcgact
     4201 acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta
     4261 ccgccaagta cttcttctac agcaacatca tgaacttttt caagaccgag attaccctgg
     4321 ccaacggcga gatccggaag cggcctctga tcgagacaaa cggcgaaacc ggggagatcg
     4381 tgtgggataa gggccgggat tttgccaccg tgcggaaagt gctgagcatg ccccaagtga
     4441 atatcgtgaa aaagaccgag gtgcagacag gcggcttcag caaagagtct atcctgccca
     4501 agaggaacag cgataagctg atcgccagaa agaaggactg ggaccctaag aagtacggcg
     4561 gcttcgacag ccccaccgtg gcctattctg tgctggtggt ggccaaagtg gaaaagggca
     4621 agtccaagaa actgaagagt gtgaaagagc tgctggggat caccatcatg gaaagaagca
     4681 gcttcgagaa gaatcccatc gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg
     4741 acctgatcat caagctgcct aagtactccc tgttcgagct ggaaaacggc cggaagagaa
     4801 tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact ggccctgccc tccaaatatg
     4861 tgaacttcct gtacctggcc agccactatg agaagctgaa gggctccccc gaggataatg
     4921 agcagaaaca gctgtttgtg gaacagcaca agcactacct ggacgagatc atcgagcaga
     4981 tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa tctggacaaa gtgctgtccg
     5041 cctacaacaa gcaccgggat aagcccatca gagagcaggc cgagaatatc atccacctgt
     5101 ttaccctgac caatctggga gcccctgccg ccttcaagta ctttgacacc accatcgacc
     5161 ggaagaggta caccagcacc aaagaggtgc tggacgccac cctgatccac cagagcatca
     5221 ccggcctgta cgagacacgg atcgacctgt ctcagctggg aggcgacaaa aggccggcgg
     5281 ccacgaaaaa ggccggccag gcaaaaaaga aaaaggaatt cggcagtgga gagggcagag
     5341 gaagtctgct aacatgcggt gacgtcgagg agaatcctgg cccagtgagc aagggcgagg
     5401 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     5461 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     5521 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     5581 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     5641 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     5701 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     5761 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     5821 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     5881 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     5941 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     6001 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     6061 ccgccgggat cactctcggc atggacgagc tgtacaagga attctaacta gagctcgctg
     6121 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     6181 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     6241 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     6301 gggggaggat tgggaagaga atagcaggca tgctggggag cggccgcagg aacccctagt
     6361 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     6421 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
     6481 cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     6541 catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     6601 gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct
     6661 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     6721 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg
     6781 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     6841 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc
     6901 tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat
     6961 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta
     7021 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     7081 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     7141 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     7201 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     7261 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     7321 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     7381 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     7441 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     7501 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     7561 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     7621 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     7681 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     7741 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     7801 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     7861 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     7921 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     7981 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     8041 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     8101 tctggagccg gtgagcgtgg aagccgcggt atcattgcag cactggggcc agatggtaag
     8161 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     8221 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     8281 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     8341 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     8401 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     8461 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     8521 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     8581 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     8641 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     8701 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     8761 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     8821 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     8881 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     8941 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     9001 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     9061 ttttgctggc cttttgctca catgtgaggg cctatttccc atgattcctt catatttgca
     9121 tatacgatac aaggctgtta gagagataat tggaattaat ttgactgtaa acacaaagat
     9181 attagtacaa aatacgtgac gtagaaagta ataatttctt gggtagtttg cagttttaaa
     9241 attatgtttt aaaatggact atcatatgct taccgtaact tgaaagtatt
//
