LOCUS       sequence-364642-        9245 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes gRNA for mouse TRIM28 along with Cas9 with 2A-EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9245)
  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 9245)
  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-GFP-TRIM28
FEATURES             Location/Qualifiers
     source          1..9245
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          117..344
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      356..365
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             365..430
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             437..457
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             482..4582
                     /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             482..4582
                     /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             4583..4630
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4646..4699
                     /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(4700..4702,4703..5413)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(4700..4702,4703..5413)
                     /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(5003..5022)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5350..5371
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5441..5458)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5447..5654
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5663..5803
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5663..5792
                     /label=AAV2 ITR
     rep_origin      5878..6333
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5965..5984)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6175..6196
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6350..6369)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6469..6491
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6529..6547)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6615..6719
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6720..6788,6789..7580)
                     /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(6938..6957)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7751..8339
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8240..8259
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8401..8649
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8401..8421
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8572..8591
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8671..8746
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8843..9128
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt
       61 ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag
      121 tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc
      181 ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg
      241 gctgtaatta gctgagcaag aggtaagggt ttaagggatg gttggttggt ggggtattaa
      301 tgtttaatta cctggagcac ctgcctgaaa tcactttttt tcaggttgga ccggtgccac
      361 catggactat aaggaccacg acggagacta caaggatcat gatattgatt acaaagacga
      421 tgacgataag atggccccaa agaagaagcg gaaggtcggt atccacggag tcccagcagc
      481 cgacaagaag tacagcatcg gcctggacat cggcaccaac tctgtgggct gggccgtgat
      541 caccgacgag tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca
      601 cagcatcaag aagaacctga tcggagccct gctgttcgac agcggcgaaa cagccgaggc
      661 cacccggctg aagagaaccg ccagaagaag atacaccaga cggaagaacc ggatctgcta
      721 tctgcaagag atcttcagca acgagatggc caaggtggac gacagcttct tccacagact
      781 ggaagagtcc ttcctggtgg aagaggataa gaagcacgag cggcacccca tcttcggcaa
      841 catcgtggac gaggtggcct accacgagaa gtaccccacc atctaccacc tgagaaagaa
      901 actggtggac agcaccgaca aggccgacct gcggctgatc tatctggccc tggcccacat
      961 gatcaagttc cggggccact tcctgatcga gggcgacctg aaccccgaca acagcgacgt
     1021 ggacaagctg ttcatccagc tggtgcagac ctacaaccag ctgttcgagg aaaaccccat
     1081 caacgccagc ggcgtggacg ccaaggccat cctgtctgcc agactgagca agagcagacg
     1141 gctggaaaat ctgatcgccc agctgcccgg cgagaagaag aatggcctgt tcggaaacct
     1201 gattgccctg agcctgggcc tgacccccaa cttcaagagc aacttcgacc tggccgagga
     1261 tgccaaactg cagctgagca aggacaccta cgacgacgac ctggacaacc tgctggccca
     1321 gatcggcgac cagtacgccg acctgtttct ggccgccaag aacctgtccg acgccatcct
     1381 gctgagcgac atcctgagag tgaacaccga gatcaccaag gcccccctga gcgcctctat
     1441 gatcaagaga tacgacgagc accaccagga cctgaccctg ctgaaagctc tcgtgcggca
     1501 gcagctgcct gagaagtaca aagagatttt cttcgaccag agcaagaacg gctacgccgg
     1561 ctacattgac ggcggagcca gccaggaaga gttctacaag ttcatcaagc ccatcctgga
     1621 aaagatggac ggcaccgagg aactgctcgt gaagctgaac agagaggacc tgctgcggaa
     1681 gcagcggacc ttcgacaacg gcagcatccc ccaccagatc cacctgggag agctgcacgc
     1741 cattctgcgg cggcaggaag atttttaccc attcctgaag gacaaccggg aaaagatcga
     1801 gaagatcctg accttccgca tcccctacta cgtgggccct ctggccaggg gaaacagcag
     1861 attcgcctgg atgaccagaa agagcgagga aaccatcacc ccctggaact tcgaggaagt
     1921 ggtggacaag ggcgcttccg cccagagctt catcgagcgg atgaccaact tcgataagaa
     1981 cctgcccaac gagaaggtgc tgcccaagca cagcctgctg tacgagtact tcaccgtgta
     2041 taacgagctg accaaagtga aatacgtgac cgagggaatg agaaagcccg ccttcctgag
     2101 cggcgagcag aaaaaggcca tcgtggacct gctgttcaag accaaccgga aagtgaccgt
     2161 gaagcagctg aaagaggact acttcaagaa aatcgagtgc ttcgactccg tggaaatctc
     2221 cggcgtggaa gatcggttca acgcctccct gggcacatac cacgatctgc tgaaaattat
     2281 caaggacaag gacttcctgg acaatgagga aaacgaggac attctggaag atatcgtgct
     2341 gaccctgaca ctgtttgagg acagagagat gatcgaggaa cggctgaaaa cctatgccca
     2401 cctgttcgac gacaaagtga tgaagcagct gaagcggcgg agatacaccg gctggggcag
     2461 gctgagccgg aagctgatca acggcatccg ggacaagcag tccggcaaga caatcctgga
     2521 tttcctgaag tccgacggct tcgccaacag aaacttcatg cagctgatcc acgacgacag
     2581 cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg atagcctgca
     2641 cgagcacatt gccaatctgg ccggcagccc cgccattaag aagggcatcc tgcagacagt
     2701 gaaggtggtg gacgagctcg tgaaagtgat gggccggcac aagcccgaga acatcgtgat
     2761 cgaaatggcc agagagaacc agaccaccca gaagggacag aagaacagcc gcgagagaat
     2821 gaagcggatc gaagagggca tcaaagagct gggcagccag atcctgaaag aacaccccgt
     2881 ggaaaacacc cagctgcaga acgagaagct gtacctgtac tacctgcaga atgggcggga
     2941 tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc gactacgatg tggaccatat
     3001 cgtgcctcag agctttctga aggacgactc catcgacaac aaggtgctga ccagaagcga
     3061 caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa
     3121 ctactggcgg cagctgctga acgccaagct gattacccag agaaagttcg acaatctgac
     3181 caaggccgag agaggcggcc tgagcgaact ggataaggcc ggcttcatca agagacagct
     3241 ggtggaaacc cggcagatca caaagcacgt ggcacagatc ctggactccc ggatgaacac
     3301 taagtacgac gagaatgaca agctgatccg ggaagtgaaa gtgatcaccc tgaagtccaa
     3361 gctggtgtcc gatttccgga aggatttcca gttttacaaa gtgcgcgaga tcaacaacta
     3421 ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga accgccctga tcaaaaagta
     3481 ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat
     3541 gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc aagtacttct tctacagcaa
     3601 catcatgaac tttttcaaga ccgagattac cctggccaac ggcgagatcc ggaagcggcc
     3661 tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg gataagggcc gggattttgc
     3721 caccgtgcgg aaagtgctga gcatgcccca agtgaatatc gtgaaaaaga ccgaggtgca
     3781 gacaggcggc ttcagcaaag agtctatcct gcccaagagg aacagcgata agctgatcgc
     3841 cagaaagaag gactgggacc ctaagaagta cggcggcttc gacagcccca ccgtggccta
     3901 ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc aagaaactga agagtgtgaa
     3961 agagctgctg gggatcacca tcatggaaag aagcagcttc gagaagaatc ccatcgactt
     4021 tctggaagcc aagggctaca aagaagtgaa aaaggacctg atcatcaagc tgcctaagta
     4081 ctccctgttc gagctggaaa acggccggaa gagaatgctg gcctctgccg gcgaactgca
     4141 gaagggaaac gaactggccc tgccctccaa atatgtgaac ttcctgtacc tggccagcca
     4201 ctatgagaag ctgaagggct cccccgagga taatgagcag aaacagctgt ttgtggaaca
     4261 gcacaagcac tacctggacg agatcatcga gcagatcagc gagttctcca agagagtgat
     4321 cctggccgac gctaatctgg acaaagtgct gtccgcctac aacaagcacc gggataagcc
     4381 catcagagag caggccgaga atatcatcca cctgtttacc ctgaccaatc tgggagcccc
     4441 tgccgccttc aagtactttg acaccaccat cgaccggaag aggtacacca gcaccaaaga
     4501 ggtgctggac gccaccctga tccaccagag catcaccggc ctgtacgaga cacggatcga
     4561 cctgtctcag ctgggaggcg acaaaaggcc ggcggccacg aaaaaggccg gccaggcaaa
     4621 aaagaaaaag gaattcggca gtggagaggg cagaggaagt ctgctaacat gcggtgacgt
     4681 cgaggagaat cctggcccag tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     4741 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     4801 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     4861 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     4921 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     4981 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     5041 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     5101 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     5161 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     5221 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     5281 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     5341 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     5401 cgagctgtac aaggaattct aactagagct cgctgatcag cctcgactgt gccttctagt
     5461 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     5521 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     5581 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agagaatagc
     5641 aggcatgctg gggagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     5701 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     5761 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
     5821 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     5881 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     5941 cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     6001 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     6061 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     6121 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     6181 tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt gatttataag
     6241 ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     6301 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     6361 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     6421 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     6481 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     6541 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     6601 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     6661 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     6721 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     6781 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     6841 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     6901 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     6961 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     7021 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     7081 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     7141 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     7201 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     7261 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     7321 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     7381 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtggaagcc
     7441 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     7501 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     7561 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     7621 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     7681 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     7741 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     7801 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     7861 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     7921 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     7981 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     8041 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     8101 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     8161 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     8221 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     8281 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     8341 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     8401 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
     8461 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
     8521 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
     8581 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
     8641 cgaaacaccg cgtgtgaatg gcggcctcgg gttttagagc tagaaatagc aagttaaaat
     8701 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgttttaga
     8761 gctagaaata gcaagttaaa ataaggctag tccgttttta gcgcgtgcgc caattctgca
     8821 gacaaatggc tctagaggta cccgttacat aacttacggt aaatggcccg cctggctgac
     8881 cgcccaacga cccccgccca ttgacgtcaa tagtaacgcc aatagggact ttccattgac
     8941 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     9001 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattgtgccc
     9061 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     9121 ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc cccccctccc
     9181 cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg ggggcggggg
     9241 ggggg
//
