LOCUS       sequence-364582-        9450 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 9450)
  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 9450)
  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-MMETn-del-gRNA1
FEATURES             Location/Qualifiers
     source          1..9450
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gap             26..48
                     /estimated_length=23
     intron          182..409
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      421..430
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             430..495
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             502..522
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             547..4647
                     /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             547..4647
                     /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             4648..4695
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4711..4764
                     /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(4765..4767,4768..5478)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(4765..4767,4768..5478)
                     /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(5068..5087)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5415..5436
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5506..5523)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5512..5719
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5728..5868
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5728..5857
                     /label=AAV2 ITR
     rep_origin      5943..6398
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6030..6049)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6240..6261
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6415..6434)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6534..6556
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6594..6612)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6680..6784
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6785..6853,6854..7645)
                     /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(7003..7022)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7816..8404
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8305..8324
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8466..8714
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8466..8486
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8637..8656
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8736..8811
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8908..9193
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 gggggggggg gggggggggg gggggnnnnn nnnnnnnnnn nnnnnnnngg gngnggggcg
       61 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      121 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      181 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      241 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      301 tccgggctgt aattagctga gcaagaggta agggtttaag ggatggttgg ttggtggggt
      361 attaatgttt aattacctgg agcacctgcc tgaaatcact ttttttcagg ttggaccggt
      421 gccaccatgg actataagga ccacgacgga gactacaagg atcatgatat tgattacaaa
      481 gacgatgacg ataagatggc cccaaagaag aagcggaagg tcggtatcca cggagtccca
      541 gcagccgaca agaagtacag catcggcctg gacatcggca ccaactctgt gggctgggcc
      601 gtgatcaccg acgagtacaa ggtgcccagc aagaaattca aggtgctggg caacaccgac
      661 cggcacagca tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc
      721 gaggccaccc ggctgaagag aaccgccaga agaagataca ccagacggaa gaaccggatc
      781 tgctatctgc aagagatctt cagcaacgag atggccaagg tggacgacag cttcttccac
      841 agactggaag agtccttcct ggtggaagag gataagaagc acgagcggca ccccatcttc
      901 ggcaacatcg tggacgaggt ggcctaccac gagaagtacc ccaccatcta ccacctgaga
      961 aagaaactgg tggacagcac cgacaaggcc gacctgcggc tgatctatct ggccctggcc
     1021 cacatgatca agttccgggg ccacttcctg atcgagggcg acctgaaccc cgacaacagc
     1081 gacgtggaca agctgttcat ccagctggtg cagacctaca accagctgtt cgaggaaaac
     1141 cccatcaacg ccagcggcgt ggacgccaag gccatcctgt ctgccagact gagcaagagc
     1201 agacggctgg aaaatctgat cgcccagctg cccggcgaga agaagaatgg cctgttcgga
     1261 aacctgattg ccctgagcct gggcctgacc cccaacttca agagcaactt cgacctggcc
     1321 gaggatgcca aactgcagct gagcaaggac acctacgacg acgacctgga caacctgctg
     1381 gcccagatcg gcgaccagta cgccgacctg tttctggccg ccaagaacct gtccgacgcc
     1441 atcctgctga gcgacatcct gagagtgaac accgagatca ccaaggcccc cctgagcgcc
     1501 tctatgatca agagatacga cgagcaccac caggacctga ccctgctgaa agctctcgtg
     1561 cggcagcagc tgcctgagaa gtacaaagag attttcttcg accagagcaa gaacggctac
     1621 gccggctaca ttgacggcgg agccagccag gaagagttct acaagttcat caagcccatc
     1681 ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg
     1741 cggaagcagc ggaccttcga caacggcagc atcccccacc agatccacct gggagagctg
     1801 cacgccattc tgcggcggca ggaagatttt tacccattcc tgaaggacaa ccgggaaaag
     1861 atcgagaaga tcctgacctt ccgcatcccc tactacgtgg gccctctggc caggggaaac
     1921 agcagattcg cctggatgac cagaaagagc gaggaaacca tcaccccctg gaacttcgag
     1981 gaagtggtgg acaagggcgc ttccgcccag agcttcatcg agcggatgac caacttcgat
     2041 aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga gtacttcacc
     2101 gtgtataacg agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc
     2161 ctgagcggcg agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg
     2221 accgtgaagc agctgaaaga ggactacttc aagaaaatcg agtgcttcga ctccgtggaa
     2281 atctccggcg tggaagatcg gttcaacgcc tccctgggca cataccacga tctgctgaaa
     2341 attatcaagg acaaggactt cctggacaat gaggaaaacg aggacattct ggaagatatc
     2401 gtgctgaccc tgacactgtt tgaggacaga gagatgatcg aggaacggct gaaaacctat
     2461 gcccacctgt tcgacgacaa agtgatgaag cagctgaagc ggcggagata caccggctgg
     2521 ggcaggctga gccggaagct gatcaacggc atccgggaca agcagtccgg caagacaatc
     2581 ctggatttcc tgaagtccga cggcttcgcc aacagaaact tcatgcagct gatccacgac
     2641 gacagcctga cctttaaaga ggacatccag aaagcccagg tgtccggcca gggcgatagc
     2701 ctgcacgagc acattgccaa tctggccggc agccccgcca ttaagaaggg catcctgcag
     2761 acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc
     2821 gtgatcgaaa tggccagaga gaaccagacc acccagaagg gacagaagaa cagccgcgag
     2881 agaatgaagc ggatcgaaga gggcatcaaa gagctgggca gccagatcct gaaagaacac
     2941 cccgtggaaa acacccagct gcagaacgag aagctgtacc tgtactacct gcagaatggg
     3001 cgggatatgt acgtggacca ggaactggac atcaaccggc tgtccgacta cgatgtggac
     3061 catatcgtgc ctcagagctt tctgaaggac gactccatcg acaacaaggt gctgaccaga
     3121 agcgacaaga accggggcaa gagcgacaac gtgccctccg aagaggtcgt gaagaagatg
     3181 aagaactact ggcggcagct gctgaacgcc aagctgatta cccagagaaa gttcgacaat
     3241 ctgaccaagg ccgagagagg cggcctgagc gaactggata aggccggctt catcaagaga
     3301 cagctggtgg aaacccggca gatcacaaag cacgtggcac agatcctgga ctcccggatg
     3361 aacactaagt acgacgagaa tgacaagctg atccgggaag tgaaagtgat caccctgaag
     3421 tccaagctgg tgtccgattt ccggaaggat ttccagtttt acaaagtgcg cgagatcaac
     3481 aactaccacc acgcccacga cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa
     3541 aagtacccta agctggaaag cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg
     3601 aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta cttcttctac
     3661 agcaacatca tgaacttttt caagaccgag attaccctgg ccaacggcga gatccggaag
     3721 cggcctctga tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa gggccgggat
     3781 tttgccaccg tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa aaagaccgag
     3841 gtgcagacag gcggcttcag caaagagtct atcctgccca agaggaacag cgataagctg
     3901 atcgccagaa agaaggactg ggaccctaag aagtacggcg gcttcgacag ccccaccgtg
     3961 gcctattctg tgctggtggt ggccaaagtg gaaaagggca agtccaagaa actgaagagt
     4021 gtgaaagagc tgctggggat caccatcatg gaaagaagca gcttcgagaa gaatcccatc
     4081 gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat caagctgcct
     4141 aagtactccc tgttcgagct ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa
     4201 ctgcagaagg gaaacgaact ggccctgccc tccaaatatg tgaacttcct gtacctggcc
     4261 agccactatg agaagctgaa gggctccccc gaggataatg agcagaaaca gctgtttgtg
     4321 gaacagcaca agcactacct ggacgagatc atcgagcaga tcagcgagtt ctccaagaga
     4381 gtgatcctgg ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa gcaccgggat
     4441 aagcccatca gagagcaggc cgagaatatc atccacctgt ttaccctgac caatctggga
     4501 gcccctgccg ccttcaagta ctttgacacc accatcgacc ggaagaggta caccagcacc
     4561 aaagaggtgc tggacgccac cctgatccac cagagcatca ccggcctgta cgagacacgg
     4621 atcgacctgt ctcagctggg aggcgacaaa aggccggcgg ccacgaaaaa ggccggccag
     4681 gcaaaaaaga aaaaggaatt cggcagtgga gagggcagag gaagtctgct aacatgcggt
     4741 gacgtcgagg agaatcctgg cccagtgagc aagggcgagg agctgttcac cggggtggtg
     4801 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     4861 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     4921 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     4981 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     5041 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     5101 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     5161 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     5221 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     5281 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     5341 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     5401 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     5461 atggacgagc tgtacaagga attctaacta gagctcgctg atcagcctcg actgtgcctt
     5521 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     5581 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     5641 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaga
     5701 atagcaggca tgctggggag cggccgcagg aacccctagt gatggagttg gccactccct
     5761 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     5821 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc
     5881 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     5941 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     6001 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     6061 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     6121 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     6181 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     6241 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt
     6301 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt
     6361 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     6421 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     6481 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     6541 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     6601 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     6661 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     6721 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     6781 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     6841 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     6901 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     6961 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     7021 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     7081 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     7141 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     7201 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     7261 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     7321 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     7381 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     7441 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     7501 aagccgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     7561 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     7621 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     7681 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     7741 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     7801 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     7861 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     7921 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     7981 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     8041 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     8101 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     8161 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     8221 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     8281 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     8341 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     8401 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     8461 catgtgaggg cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta
     8521 gagagataat tggaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac
     8581 gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact
     8641 atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga
     8701 aaggacgaaa caccgaaaca tcagtgaaac agagtgtttt agagctagaa atagcaagtt
     8761 aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg cttttttgtt
     8821 ttagagctag aaatagcaag ttaaaataag gctagtccgt ttttagcgcg tgcgccaatt
     8881 ctgcagacaa atggctctag aggtacccgt tacataactt acggtaaatg gcccgcctgg
     8941 ctgaccgccc aacgaccccc gcccattgac gtcaatagta acgccaatag ggactttcca
     9001 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     9061 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcattg
     9121 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     9181 cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca tctccccccc
     9241 ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag cgatgggggc
     9301 gggggggggg ggggggscss sccvsggggg gggggggggg gggggggggg gggggggggg
     9361 gggggggggg gggggggggg gggggngngg gggggggggg ngggsggggg ggggkggggg
     9421 gggggggggg gggggggggg gggggggggg
//
