LOCUS       sequence-364644-        9258 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of gRNA and Cas9 from S. pyogenes with 2A-EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9258)
  AUTHORS   Asimi V, Sampath Kumar A, Niskanen H, Riemenschneider C, Hetzel S, 
            Naderi J, Fasching N, Popitsch N, Du M, Kretzmer H, Smith ZD, 
            Weigert R, Walther M, Mamde S, Meierhofer D, Wittler L, Buschow R, 
            Timmermann B, Cisse II, Ameres SL, Meissner A, Hnisz D
  TITLE     Hijacking of transcriptional condensates by endogenous retroviruses.
  JOURNAL   Nat Genet. 2022 Aug;54(8):1238-1247. doi: 
            10.1038/s41588-022-01132-w. Epub 2022 Jul 21.
  PUBMED    35864192
REFERENCE   2  (bases 1 to 9258)
  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: 
            "10.1038/s41588-022-01132-w"; journalName: "Nat Genet"; date: 
            "2022-08"; volume: "54"; issue: "8"; pages: "1238-1247"
COMMENT     Sequence Label: pX458-MMERVK9C-del-gRNA1
FEATURES             Location/Qualifiers
     source          1..9258
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          129..356
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      368..377
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             377..442
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             449..469
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             494..4594
                     /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             494..4594
                     /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             4595..4642
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4658..4711
                     /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(4712..4714,4715..5425)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(4712..4714,4715..5425)
                     /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(5015..5034)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5362..5383
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5453..5470)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5459..5666
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5675..5815
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5675..5804
                     /label=AAV2 ITR
     rep_origin      5890..6345
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5977..5996)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6187..6208
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6362..6381)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6481..6503
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6541..6559)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6627..6731
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6732..6800,6801..7592)
                     /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(6950..6969)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7763..8351
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8252..8271
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8413..8661
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8413..8433
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8584..8603
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8683..8758
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8855..9140
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 aggggcgggg cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc
       61 gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc
      121 ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc
      181 gccgcccgcc ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc
      241 cttctcctcc gggctgtaat tagctgagca agaggtaagg gtttaaggga tggttggttg
      301 gtggggtatt aatgtttaat tacctggagc acctgcctga aatcactttt tttcaggttg
      361 gaccggtgcc accatggact ataaggacca cgacggagac tacaaggatc atgatattga
      421 ttacaaagac gatgacgata agatggcccc aaagaagaag cggaaggtcg gtatccacgg
      481 agtcccagca gccgacaaga agtacagcat cggcctggac atcggcacca actctgtggg
      541 ctgggccgtg atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa
      601 caccgaccgg cacagcatca agaagaacct gatcggagcc ctgctgttcg acagcggcga
      661 aacagccgag gccacccggc tgaagagaac cgccagaaga agatacacca gacggaagaa
      721 ccggatctgc tatctgcaag agatcttcag caacgagatg gccaaggtgg acgacagctt
      781 cttccacaga ctggaagagt ccttcctggt ggaagaggat aagaagcacg agcggcaccc
      841 catcttcggc aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca
      901 cctgagaaag aaactggtgg acagcaccga caaggccgac ctgcggctga tctatctggc
      961 cctggcccac atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga
     1021 caacagcgac gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga
     1081 ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg ccagactgag
     1141 caagagcaga cggctggaaa atctgatcgc ccagctgccc ggcgagaaga agaatggcct
     1201 gttcggaaac ctgattgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga
     1261 cctggccgag gatgccaaac tgcagctgag caaggacacc tacgacgacg acctggacaa
     1321 cctgctggcc cagatcggcg accagtacgc cgacctgttt ctggccgcca agaacctgtc
     1381 cgacgccatc ctgctgagcg acatcctgag agtgaacacc gagatcacca aggcccccct
     1441 gagcgcctct atgatcaaga gatacgacga gcaccaccag gacctgaccc tgctgaaagc
     1501 tctcgtgcgg cagcagctgc ctgagaagta caaagagatt ttcttcgacc agagcaagaa
     1561 cggctacgcc ggctacattg acggcggagc cagccaggaa gagttctaca agttcatcaa
     1621 gcccatcctg gaaaagatgg acggcaccga ggaactgctc gtgaagctga acagagagga
     1681 cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg
     1741 agagctgcac gccattctgc ggcggcagga agatttttac ccattcctga aggacaaccg
     1801 ggaaaagatc gagaagatcc tgaccttccg catcccctac tacgtgggcc ctctggccag
     1861 gggaaacagc agattcgcct ggatgaccag aaagagcgag gaaaccatca ccccctggaa
     1921 cttcgaggaa gtggtggaca agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa
     1981 cttcgataag aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta
     2041 cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc
     2101 cgccttcctg agcggcgagc agaaaaaggc catcgtggac ctgctgttca agaccaaccg
     2161 gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt gcttcgactc
     2221 cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc ctgggcacat accacgatct
     2281 gctgaaaatt atcaaggaca aggacttcct ggacaatgag gaaaacgagg acattctgga
     2341 agatatcgtg ctgaccctga cactgtttga ggacagagag atgatcgagg aacggctgaa
     2401 aacctatgcc cacctgttcg acgacaaagt gatgaagcag ctgaagcggc ggagatacac
     2461 cggctggggc aggctgagcc ggaagctgat caacggcatc cgggacaagc agtccggcaa
     2521 gacaatcctg gatttcctga agtccgacgg cttcgccaac agaaacttca tgcagctgat
     2581 ccacgacgac agcctgacct ttaaagagga catccagaaa gcccaggtgt ccggccaggg
     2641 cgatagcctg cacgagcaca ttgccaatct ggccggcagc cccgccatta agaagggcat
     2701 cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc acaagcccga
     2761 gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc cagaagggac agaagaacag
     2821 ccgcgagaga atgaagcgga tcgaagaggg catcaaagag ctgggcagcc agatcctgaa
     2881 agaacacccc gtggaaaaca cccagctgca gaacgagaag ctgtacctgt actacctgca
     2941 gaatgggcgg gatatgtacg tggaccagga actggacatc aaccggctgt ccgactacga
     3001 tgtggaccat atcgtgcctc agagctttct gaaggacgac tccatcgaca acaaggtgct
     3061 gaccagaagc gacaagaacc ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa
     3121 gaagatgaag aactactggc ggcagctgct gaacgccaag ctgattaccc agagaaagtt
     3181 cgacaatctg accaaggccg agagaggcgg cctgagcgaa ctggataagg ccggcttcat
     3241 caagagacag ctggtggaaa cccggcagat cacaaagcac gtggcacaga tcctggactc
     3301 ccggatgaac actaagtacg acgagaatga caagctgatc cgggaagtga aagtgatcac
     3361 cctgaagtcc aagctggtgt ccgatttccg gaaggatttc cagttttaca aagtgcgcga
     3421 gatcaacaac taccaccacg cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct
     3481 gatcaaaaag taccctaagc tggaaagcga gttcgtgtac ggcgactaca aggtgtacga
     3541 cgtgcggaag atgatcgcca agagcgagca ggaaatcggc aaggctaccg ccaagtactt
     3601 cttctacagc aacatcatga actttttcaa gaccgagatt accctggcca acggcgagat
     3661 ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt gggataaggg
     3721 ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa
     3781 gaccgaggtg cagacaggcg gcttcagcaa agagtctatc ctgcccaaga ggaacagcga
     3841 taagctgatc gccagaaaga aggactggga ccctaagaag tacggcggct tcgacagccc
     3901 caccgtggcc tattctgtgc tggtggtggc caaagtggaa aagggcaagt ccaagaaact
     3961 gaagagtgtg aaagagctgc tggggatcac catcatggaa agaagcagct tcgagaagaa
     4021 tcccatcgac tttctggaag ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa
     4081 gctgcctaag tactccctgt tcgagctgga aaacggccgg aagagaatgc tggcctctgc
     4141 cggcgaactg cagaagggaa acgaactggc cctgccctcc aaatatgtga acttcctgta
     4201 cctggccagc cactatgaga agctgaaggg ctcccccgag gataatgagc agaaacagct
     4261 gtttgtggaa cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttctc
     4321 caagagagtg atcctggccg acgctaatct ggacaaagtg ctgtccgcct acaacaagca
     4381 ccgggataag cccatcagag agcaggccga gaatatcatc cacctgttta ccctgaccaa
     4441 tctgggagcc cctgccgcct tcaagtactt tgacaccacc atcgaccgga agaggtacac
     4501 cagcaccaaa gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga
     4561 gacacggatc gacctgtctc agctgggagg cgacaaaagg ccggcggcca cgaaaaaggc
     4621 cggccaggca aaaaagaaaa aggaattcgg cagtggagag ggcagaggaa gtctgctaac
     4681 atgcggtgac gtcgaggaga atcctggccc agtgagcaag ggcgaggagc tgttcaccgg
     4741 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     4801 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     4861 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
     4921 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     4981 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     5041 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     5101 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     5161 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
     5221 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     5281 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga
     5341 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     5401 tctcggcatg gacgagctgt acaaggaatt ctaactagag ctcgctgatc agcctcgact
     5461 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     5521 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     5581 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     5641 gaagagaata gcaggcatgc tggggagcgg ccgcaggaac ccctagtgat ggagttggcc
     5701 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
     5761 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc
     5821 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc
     5881 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     5941 gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct
     6001 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     6061 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     6121 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     6181 ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt
     6241 ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac
     6301 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca
     6361 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     6421 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     6481 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     6541 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     6601 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     6661 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     6721 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     6781 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     6841 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     6901 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     6961 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     7021 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     7081 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     7141 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     7201 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     7261 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     7321 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     7381 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     7441 agcgtggaag ccgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     7501 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     7561 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     7621 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     7681 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     7741 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     7801 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     7861 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     7921 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     7981 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     8041 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     8101 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     8161 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     8221 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     8281 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     8341 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     8401 ttgctcacat gtgagggcct atttcccatg attccttcat atttgcatat acgatacaag
     8461 gctgttagag agataattgg aattaatttg actgtaaaca caaagatatt agtacaaaat
     8521 acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa
     8581 atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc
     8641 ttgtggaaag gacgaaacac cgatatgtct gtaatcacac acgttttaga gctagaaata
     8701 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
     8761 ttttgtttta gagctagaaa tagcaagtta aaataaggct agtccgtttt tagcgcgtgc
     8821 gccaattctg cagacaaatg gctctagagg tacccgttac ataacttacg gtaaatggcc
     8881 cgcctggctg accgcccaac gacccccgcc cattgacgtc aatagtaacg ccaataggga
     8941 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     9001 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     9061 ggcattgtgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     9121 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
     9181 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
     9241 tgggggcggg gggggggg
//
