LOCUS       sequence-364581-        9290 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 9290)
  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 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: 
            "10.1038/s41588-022-01132-w"; journalName: "Nat Genet"; date: 
            "2022-08"; volume: "54"; issue: "8"; pages: "1238-1247"
COMMENT     Sequence Label: pX458-MMERVK9C-del-gRNA2
FEATURES             Location/Qualifiers
     source          1..9290
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          111..338
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      350..359
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             359..424
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             431..451
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             476..4576
                     /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             476..4576
                     /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             4577..4624
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4640..4693
                     /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(4694..4696,4697..5407)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(4694..4696,4697..5407)
                     /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(4997..5016)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5344..5365
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5435..5452)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5441..5648
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5657..5797
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5657..5786
                     /label=AAV2 ITR
     rep_origin      5872..6327
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5959..5978)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6169..6190
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6344..6363)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6463..6485
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6523..6541)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6609..6713
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6714..6782,6783..7574)
                     /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(6932..6951)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7745..8333
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8234..8253
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8395..8643
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8395..8415
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8566..8585
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8665..8740
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8837..9122
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     gap             9242..9267
                     /estimated_length=26
ORIGIN
        1 ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg
       61 cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg
      121 cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct
      181 gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta
      241 attagctgag caagaggtaa gggtttaagg gatggttggt tggtggggta ttaatgttta
      301 attacctgga gcacctgcct gaaatcactt tttttcaggt tggaccggtg ccaccatgga
      361 ctataaggac cacgacggag actacaagga tcatgatatt gattacaaag acgatgacga
      421 taagatggcc ccaaagaaga agcggaaggt cggtatccac ggagtcccag cagccgacaa
      481 gaagtacagc atcggcctgg acatcggcac caactctgtg ggctgggccg tgatcaccga
      541 cgagtacaag gtgcccagca agaaattcaa ggtgctgggc aacaccgacc ggcacagcat
      601 caagaagaac ctgatcggag ccctgctgtt cgacagcggc gaaacagccg aggccacccg
      661 gctgaagaga accgccagaa gaagatacac cagacggaag aaccggatct gctatctgca
      721 agagatcttc agcaacgaga tggccaaggt ggacgacagc ttcttccaca gactggaaga
      781 gtccttcctg gtggaagagg ataagaagca cgagcggcac cccatcttcg gcaacatcgt
      841 ggacgaggtg gcctaccacg agaagtaccc caccatctac cacctgagaa agaaactggt
      901 ggacagcacc gacaaggccg acctgcggct gatctatctg gccctggccc acatgatcaa
      961 gttccggggc cacttcctga tcgagggcga cctgaacccc gacaacagcg acgtggacaa
     1021 gctgttcatc cagctggtgc agacctacaa ccagctgttc gaggaaaacc ccatcaacgc
     1081 cagcggcgtg gacgccaagg ccatcctgtc tgccagactg agcaagagca gacggctgga
     1141 aaatctgatc gcccagctgc ccggcgagaa gaagaatggc ctgttcggaa acctgattgc
     1201 cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc gacctggccg aggatgccaa
     1261 actgcagctg agcaaggaca cctacgacga cgacctggac aacctgctgg cccagatcgg
     1321 cgaccagtac gccgacctgt ttctggccgc caagaacctg tccgacgcca tcctgctgag
     1381 cgacatcctg agagtgaaca ccgagatcac caaggccccc ctgagcgcct ctatgatcaa
     1441 gagatacgac gagcaccacc aggacctgac cctgctgaaa gctctcgtgc ggcagcagct
     1501 gcctgagaag tacaaagaga ttttcttcga ccagagcaag aacggctacg ccggctacat
     1561 tgacggcgga gccagccagg aagagttcta caagttcatc aagcccatcc tggaaaagat
     1621 ggacggcacc gaggaactgc tcgtgaagct gaacagagag gacctgctgc ggaagcagcg
     1681 gaccttcgac aacggcagca tcccccacca gatccacctg ggagagctgc acgccattct
     1741 gcggcggcag gaagattttt acccattcct gaaggacaac cgggaaaaga tcgagaagat
     1801 cctgaccttc cgcatcccct actacgtggg ccctctggcc aggggaaaca gcagattcgc
     1861 ctggatgacc agaaagagcg aggaaaccat caccccctgg aacttcgagg aagtggtgga
     1921 caagggcgct tccgcccaga gcttcatcga gcggatgacc aacttcgata agaacctgcc
     1981 caacgagaag gtgctgccca agcacagcct gctgtacgag tacttcaccg tgtataacga
     2041 gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc tgagcggcga
     2101 gcagaaaaag gccatcgtgg acctgctgtt caagaccaac cggaaagtga ccgtgaagca
     2161 gctgaaagag gactacttca agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt
     2221 ggaagatcgg ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa ttatcaagga
     2281 caaggacttc ctggacaatg aggaaaacga ggacattctg gaagatatcg tgctgaccct
     2341 gacactgttt gaggacagag agatgatcga ggaacggctg aaaacctatg cccacctgtt
     2401 cgacgacaaa gtgatgaagc agctgaagcg gcggagatac accggctggg gcaggctgag
     2461 ccggaagctg atcaacggca tccgggacaa gcagtccggc aagacaatcc tggatttcct
     2521 gaagtccgac ggcttcgcca acagaaactt catgcagctg atccacgacg acagcctgac
     2581 ctttaaagag gacatccaga aagcccaggt gtccggccag ggcgatagcc tgcacgagca
     2641 cattgccaat ctggccggca gccccgccat taagaagggc atcctgcaga cagtgaaggt
     2701 ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg tgatcgaaat
     2761 ggccagagag aaccagacca cccagaaggg acagaagaac agccgcgaga gaatgaagcg
     2821 gatcgaagag ggcatcaaag agctgggcag ccagatcctg aaagaacacc ccgtggaaaa
     2881 cacccagctg cagaacgaga agctgtacct gtactacctg cagaatgggc gggatatgta
     2941 cgtggaccag gaactggaca tcaaccggct gtccgactac gatgtggacc atatcgtgcc
     3001 tcagagcttt ctgaaggacg actccatcga caacaaggtg ctgaccagaa gcgacaagaa
     3061 ccggggcaag agcgacaacg tgccctccga agaggtcgtg aagaagatga agaactactg
     3121 gcggcagctg ctgaacgcca agctgattac ccagagaaag ttcgacaatc tgaccaaggc
     3181 cgagagaggc ggcctgagcg aactggataa ggccggcttc atcaagagac agctggtgga
     3241 aacccggcag atcacaaagc acgtggcaca gatcctggac tcccggatga acactaagta
     3301 cgacgagaat gacaagctga tccgggaagt gaaagtgatc accctgaagt ccaagctggt
     3361 gtccgatttc cggaaggatt tccagtttta caaagtgcgc gagatcaaca actaccacca
     3421 cgcccacgac gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa
     3481 gctggaaagc gagttcgtgt acggcgacta caaggtgtac gacgtgcgga agatgatcgc
     3541 caagagcgag caggaaatcg gcaaggctac cgccaagtac ttcttctaca gcaacatcat
     3601 gaactttttc aagaccgaga ttaccctggc caacggcgag atccggaagc ggcctctgat
     3661 cgagacaaac ggcgaaaccg gggagatcgt gtgggataag ggccgggatt ttgccaccgt
     3721 gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg
     3781 cggcttcagc aaagagtcta tcctgcccaa gaggaacagc gataagctga tcgccagaaa
     3841 gaaggactgg gaccctaaga agtacggcgg cttcgacagc cccaccgtgg cctattctgt
     3901 gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct
     3961 gctggggatc accatcatgg aaagaagcag cttcgagaag aatcccatcg actttctgga
     4021 agccaagggc tacaaagaag tgaaaaagga cctgatcatc aagctgccta agtactccct
     4081 gttcgagctg gaaaacggcc ggaagagaat gctggcctct gccggcgaac tgcagaaggg
     4141 aaacgaactg gccctgccct ccaaatatgt gaacttcctg tacctggcca gccactatga
     4201 gaagctgaag ggctcccccg aggataatga gcagaaacag ctgtttgtgg aacagcacaa
     4261 gcactacctg gacgagatca tcgagcagat cagcgagttc tccaagagag tgatcctggc
     4321 cgacgctaat ctggacaaag tgctgtccgc ctacaacaag caccgggata agcccatcag
     4381 agagcaggcc gagaatatca tccacctgtt taccctgacc aatctgggag cccctgccgc
     4441 cttcaagtac tttgacacca ccatcgaccg gaagaggtac accagcacca aagaggtgct
     4501 ggacgccacc ctgatccacc agagcatcac cggcctgtac gagacacgga tcgacctgtc
     4561 tcagctggga ggcgacaaaa ggccggcggc cacgaaaaag gccggccagg caaaaaagaa
     4621 aaaggaattc ggcagtggag agggcagagg aagtctgcta acatgcggtg acgtcgagga
     4681 gaatcctggc ccagtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     4741 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     4801 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     4861 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     4921 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     4981 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     5041 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     5101 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     5161 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     5221 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     5281 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     5341 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     5401 gtacaaggaa ttctaactag agctcgctga tcagcctcga ctgtgccttc tagttgccag
     5461 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     5521 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     5581 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagagaa tagcaggcat
     5641 gctggggagc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     5701 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     5761 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     5821 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     5881 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     5941 ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     6001 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     6061 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     6121 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     6181 tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta taagggattt
     6241 tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     6301 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     6361 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     6421 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     6481 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     6541 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     6601 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     6661 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     6721 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     6781 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     6841 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     6901 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     6961 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     7021 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     7081 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     7141 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     7201 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     7261 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     7321 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     7381 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtgga agccgcggta
     7441 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     7501 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     7561 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     7621 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     7681 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     7741 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     7801 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     7861 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     7921 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     7981 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     8041 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     8101 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     8161 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     8221 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     8281 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     8341 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtgagggc
     8401 ctatttccca tgattccttc atatttgcat atacgataca aggctgttag agagataatt
     8461 ggaattaatt tgactgtaaa cacaaagata ttagtacaaa atacgtgacg tagaaagtaa
     8521 taatttcttg ggtagtttgc agttttaaaa ttatgtttta aaatggacta tcatatgctt
     8581 accgtaactt gaaagtattt cgatttcttg gctttatata tcttgtggaa aggacgaaac
     8641 accgactgtt ttgtcattct gtgggtttta gagctagaaa tagcaagtta aaataaggct
     8701 agtccgttat caacttgaaa aagtggcacc gagtcggtgc ttttttgttt tagagctaga
     8761 aatagcaagt taaaataagg ctagtccgtt tttagcgcgt gcgccaattc tgcagacaaa
     8821 tggctctaga ggtacccgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     8881 acgacccccg cccattgacg tcaatagtaa cgccaatagg gactttccat tgacgtcaat
     8941 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     9001 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattgt gcccagtaca
     9061 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     9121 tggtcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc tccccacccc
     9181 caattttgta tttatttatt ttttaattat tttgtgcagc gatgggggcg gggggggggg
     9241 gnnnnnnnnn nnnnnnnnnn nnnnnnnggg cgaggggcgg ggcggggcga
//
