LOCUS       sequence-364580-        9371 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 9371)
  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 9371)
  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-IAP-del-gRNA2
FEATURES             Location/Qualifiers
     source          1..9371
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          140..367
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      379..388
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             388..453
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             460..480
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             505..4605
                     /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             505..4605
                     /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             4606..4653
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4669..4722
                     /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(4723..4725,4726..5436)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(4723..4725,4726..5436)
                     /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(5026..5045)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5373..5394
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5464..5481)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5470..5677
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5686..5826
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5686..5815
                     /label=AAV2 ITR
     rep_origin      5901..6356
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5988..6007)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6198..6219
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6373..6392)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6492..6514
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6552..6570)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6638..6742
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6743..6811,6812..7603)
                     /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(6961..6980)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7774..8362
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8263..8282
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8424..8672
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8424..8444
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8595..8614
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8694..8769
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8866..9151
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 ggggsggggs grggggcggg gsggggcgag gcggagaggt gcggcggcag ccaatcagag
       61 cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg cggcggcggc cctataaaaa
      121 gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc
      181 cgccgcctcg cgccgcccgc cccggctctg actgaccgcg ttactcccac aggtgagcgg
      241 gcgggacggc ccttctcctc cgggctgtaa ttagctgagc aagaggtaag ggtttaaggg
      301 atggttggtt ggtggggtat taatgtttaa ttacctggag cacctgcctg aaatcacttt
      361 ttttcaggtt ggaccggtgc caccatggac tataaggacc acgacggaga ctacaaggat
      421 catgatattg attacaaaga cgatgacgat aagatggccc caaagaagaa gcggaaggtc
      481 ggtatccacg gagtcccagc agccgacaag aagtacagca tcggcctgga catcggcacc
      541 aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag
      601 gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc cctgctgttc
      661 gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag aagatacacc
      721 agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg
      781 gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac
      841 gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc
      901 accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg
      961 atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac
     1021 ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac
     1081 cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc catcctgtct
     1141 gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc cggcgagaag
     1201 aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag
     1261 agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac ctacgacgac
     1321 gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc
     1381 aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc
     1441 aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca ggacctgacc
     1501 ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac
     1561 cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga agagttctac
     1621 aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg
     1681 aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat cccccaccag
     1741 atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg
     1801 aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc
     1861 cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc
     1921 accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag cttcatcgag
     1981 cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg
     2041 ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt gaccgaggga
     2101 atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc
     2161 aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag
     2221 tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca
     2281 taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag
     2341 gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag
     2401 gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg
     2461 cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat ccgggacaag
     2521 cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc
     2581 atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggtg
     2641 tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt
     2701 aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg
     2761 cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac ccagaaggga
     2821 cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc
     2881 cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa gctgtacctg
     2941 tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat caaccggctg
     3001 tccgactacg atgtggacca tatcgtgcct cagagctttc tgaaggacga ctccatcgac
     3061 aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt gccctccgaa
     3121 gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa gctgattacc
     3181 cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga actggataag
     3241 gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca cgtggcacag
     3301 atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat ccgggaagtg
     3361 aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt ccagttttac
     3421 aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg
     3481 ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta cggcgactac
     3541 aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg caaggctacc
     3601 gccaagtact tcttctacag caacatcatg aactttttca agaccgagat taccctggcc
     3661 aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg
     3721 tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat
     3781 atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat cctgcccaag
     3841 aggaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa gtacggcggc
     3901 ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag
     3961 tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga aagaagcagc
     4021 ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt gaaaaaggac
     4081 ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg gaagagaatg
     4141 ctggcctctg ccggcgaact gcagaaggga aacgaactgg ccctgccctc caaatatgtg
     4201 aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga ggataatgag
     4261 cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat cgagcagatc
     4321 agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt gctgtccgcc
     4381 tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat ccacctgttt
     4441 accctgacca atctgggagc ccctgccgcc ttcaagtact ttgacaccac catcgaccgg
     4501 aagaggtaca ccagcaccaa agaggtgctg gacgccaccc tgatccacca gagcatcacc
     4561 ggcctgtacg agacacggat cgacctgtct cagctgggag gcgacaaaag gccggcggcc
     4621 acgaaaaagg ccggccaggc aaaaaagaaa aaggaattcg gcagtggaga gggcagagga
     4681 agtctgctaa catgcggtga cgtcgaggag aatcctggcc cagtgagcaa gggcgaggag
     4741 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     4801 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     4861 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     4921 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     4981 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     5041 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     5101 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     5161 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     5221 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     5281 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
     5341 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     5401 gccgggatca ctctcggcat ggacgagctg tacaaggaat tctaactaga gctcgctgat
     5461 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     5521 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     5581 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     5641 gggaggattg ggaagagaat agcaggcatg ctggggagcg gccgcaggaa cccctagtga
     5701 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     5761 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     5821 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     5881 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     5941 ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt
     6001 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     6061 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     6121 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     6181 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc
     6241 gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga
     6301 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     6361 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     6421 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     6481 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     6541 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     6601 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     6661 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     6721 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     6781 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     6841 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     6901 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     6961 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     7021 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     7081 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     7141 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     7201 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     7261 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     7321 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     7381 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     7441 tggagccggt gagcgtggaa gccgcggtat cattgcagca ctggggccag atggtaagcc
     7501 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     7561 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     7621 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     7681 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     7741 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     7801 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     7861 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     7921 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     7981 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     8041 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     8101 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     8161 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     8221 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     8281 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     8341 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     8401 ttgctggcct tttgctcaca tgtgagggcc tatttcccat gattccttca tatttgcata
     8461 tacgatacaa ggctgttaga gagataattg gaattaattt gactgtaaac acaaagatat
     8521 tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat
     8581 tatgttttaa aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg
     8641 ctttatatat cttgtggaaa ggacgaaaca ccggtggccg actcagtagt tgggttttag
     8701 agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg
     8761 agtcggtgct tttttgtttt agagctagaa atagcaagtt aaaataaggc tagtccgttt
     8821 ttagcgcgtg cgccaattct gcagacaaat ggctctagag gtacccgtta cataacttac
     8881 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caatagtaac
     8941 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     9001 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     9061 atggcccgcc tggcattgtg cccagtacat gaccttatgg gactttccta cttggcagta
     9121 catctacgta ttagtcatcg ctattaccat ggtcgaggtg agccccacgt tctgcttcac
     9181 tctccccatc tcccccccct ccccmccccc aattttgtat ttatttattt tttaattatt
     9241 ttgtgcagcg atgggggcgg gggggggggg ggggsgsssn cngggggggg gggggggggg
     9301 gggggggggg gggggggggg gggggggggg gggggggggg ggvsgvgncg ggggggggsg
     9361 ssgggsgggg b
//
