LOCUS       sequence-354392-       10768 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses dCas9 fused to VPR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10768)
  AUTHORS   Tak YE, Horng JE, Perry NT, Schultz HT, Iyer S, Yao Q, Zou LS, Aryee
            MJ, Pinello L, Joung JK
  TITLE     Augmenting and directing long-range CRISPR-mediated activation in 
            human cells.
  JOURNAL   Nat Methods. 2021 Sep;18(9):1075-1081. doi: 
            10.1038/s41592-021-01224-1. Epub 2021 Aug 5.
  PUBMED    34354266
REFERENCE   2  (bases 1 to 10768)
  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/s41592-021-01224-1"; journalName: "Nat Methods"; date: 
            "2021-09"; volume: "18"; issue: "9"; pages: "1075-1081"
COMMENT     Sequence Label: JEH127
FEATURES             Location/Qualifiers
     source          1..10768
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          660..1676
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1600..1622
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1684..1703
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1769..1778
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1781..1801
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1838..5236,5237..5239,5240..5836,5837,5838..5840,
                     5841..5842,5843..5845,5846..5938)
                     /codon_start=1
                     /product="Streptococcus pyogenes Cas9 endonuclease with the
                     D1135V/R1335Q/T1337R mutations"
                     /label=Cas9 VQR
                     /note="has an altered recognition specificity for the 
                     protospacer adjacent motif (PAM) (Kleinstiver et al., 
                     2015)"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(1838..1861,1862..1864,1865..4351,4352..4354,
                     4355..5938)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5945..5965
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5966..6013
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6020..6100
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             6131..6280
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             6305..6325
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6755..7111
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALL"
     CDS             7130..7699
                     /codon_start=1
                     /product="transcriptional activation domain from the human 
                     herpesvirus 4 (Epstein-Barr virus) replication and 
                     transcription activator Rta/BRLF1 (Hardwick et al., 1992; 
                     Chavez et al., 2015)"
                     /label=Rta AD
                     /translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
                     RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
                     REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
                     DTFLNDECLLHAMHISTGLSIFDTSLF"
     primer_bind     complement(7786..7805)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    7851..7906
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7905..7924)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(8267..8283)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8267..8283)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8280..8302)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8291..8307
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(8315..8321,8322..8339,8340..8345))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8360..8381
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8440..8635
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      8486..8621
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    8641..8775
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8678..8697)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8732..8751
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8843..8860)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9014..9602)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9094..9113)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9773..10564,10565..10633))
                     /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     10396..10415
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10634..10738)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      481 tgggggcggg gggggggggg gggcgcgcgc crggsggggs ggggsggggs ggggggsggg
      541 gsggggsgrg gsggrgrggk gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc
      601 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
      661 gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc
      721 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
      781 cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa
      841 gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct
      961 gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg
     1021 gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt
     1081 gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc
     1141 cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt acggggcgtg
     1201 gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg
     1261 ggccgcctcg ggccggggag ggctcggggg aggggcgcgg cggcccccgg agcgccggcg
     1321 gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc
     1381 agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc
     1441 cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg
     1501 gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc
     1561 ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga
     1621 ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc
     1681 tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattc tgcagtcgac
     1741 ggtaccgcgg gcccgggatc caccggtcgc caccatggcg ccgaaaaaaa aacgcaaagt
     1801 gaacggcgga gggtccggag gaggcggtag cggaggcgat aaaaagtatt ctattggttt
     1861 agccatcggc actaattccg ttggatgggc tgtcataacc gatgaataca aagtaccttc
     1921 aaagaaattt aaggtgttgg ggaacacaga ccgtcattcg attaaaaaga atcttatcgg
     1981 tgccctccta ttcgatagtg gcgaaacggc agaggcgact cgcctgaaac gaaccgctcg
     2041 gagaaggtat acacgtcgca agaaccgaat atgttactta caagaaattt ttagcaatga
     2101 gatggccaaa gttgacgatt ctttctttca ccgtttggaa gagtccttcc ttgtcgaaga
     2161 ggacaagaaa catgaacggc accccatctt tggaaacata gtagatgagg tggcatatca
     2221 tgaaaagtac ccaacgattt atcacctcag aaaaaagcta gttgactcaa ctgataaagc
     2281 ggacctgagg ttaatctact tggctcttgc ccatatgata aagttccgtg ggcactttct
     2341 cattgagggt gatctaaatc cggacaactc ggatgtcgac aaactgttca tccagttagt
     2401 acaaacctat aatcagttgt ttgaagagaa ccctataaat gcaagtggcg tggatgcgaa
     2461 ggctattctt agcgcccgcc tctctaaatc ccgacggcta gaaaacctga tcgcacaatt
     2521 acccggagag aagaaaaatg ggttgttcgg taaccttata gcgctctcac taggcctgac
     2581 accaaatttt aagtcgaact tcgacttagc tgaagatgcc aaattgcagc ttagtaagga
     2641 cacgtacgat gacgatctcg acaatctact ggcacaaatt ggagatcagt atgcggactt
     2701 atttttggct gccaaaaacc ttagcgatgc aatcctccta tctgacatac tgagagttaa
     2761 tactgagatt accaaggcgc cgttatccgc ttcaatgatc aaaaggtacg atgaacatca
     2821 ccaagacttg acacttctca aggccctagt ccgtcagcaa ctgcctgaga aatataagga
     2881 aatattcttt gatcagtcga aaaacgggta cgcaggttat attgacggcg gagcgagtca
     2941 agaggaattc tacaagttta tcaaacccat attagagaag atggatggga cggaagagtt
     3001 gcttgtaaaa ctcaatcgcg aagatctact gcgaaagcag cggactttcg acaacggtag
     3061 cattccacat caaatccact taggcgaatt gcatgctata cttagaaggc aggaggattt
     3121 ttatccgttc ctcaaagaca atcgtgaaaa gattgagaaa atcctaacct ttcgcatacc
     3181 ttactatgtg ggacccctgg cccgagggaa ctctcggttc gcatggatga caagaaagtc
     3241 cgaagaaacg attactccat ggaattttga ggaagttgtc gataaaggtg cgtcagctca
     3301 atcgttcatc gagaggatga ccaactttga caagaattta ccgaacgaaa aagtattgcc
     3361 taagcacagt ttactttacg agtatttcac agtgtacaat gaactcacga aagttaagta
     3421 tgtcactgag ggcatgcgta aacccgcctt tctaagcgga gaacagaaga aagcaatagt
     3481 agatctgtta ttcaagacca accgcaaagt gacagttaag caattgaaag aggactactt
     3541 taagaaaatt gaatgcttcg attctgtcga gatctccggg gtagaagatc gatttaatgc
     3601 gtcacttggt acgtatcatg acctcctaaa gataattaaa gataaggact tcctggataa
     3661 cgaagagaat gaagatatct tagaagatat agtgttgact cttaccctct ttgaagatcg
     3721 ggaaatgatt gaggaaagac taaaaacata cgctcacctg ttcgacgata aggttatgaa
     3781 acagttaaag aggcgtcgct atacgggctg gggacgattg tcgcggaaac ttatcaacgg
     3841 gataagagac aagcaaagtg gtaaaactat tctcgatttt ctaaagagcg acggcttcgc
     3901 caataggaac tttatgcagc tgatccatga tgactcttta accttcaaag aggatataca
     3961 aaaggcacag gtttccggac aaggggactc attgcacgaa catattgcga atcttgctgg
     4021 ttcgccagcc atcaaaaagg gcatactcca gacagtcaaa gtagtggatg agctagttaa
     4081 ggtcatggga cgtcacaaac cggaaaacat tgtaatcgag atggcacgcg aaaatcaaac
     4141 gactcagaag gggcaaaaaa acagtcgaga gcggatgaag agaatagaag agggtattaa
     4201 agaactgggc agccagatct taaaggagca tcctgtggaa aatacccaat tgcagaacga
     4261 gaaactttac ctctattacc tacaaaatgg aagggacatg tatgttgatc aggaactgga
     4321 cataaaccgt ttatctgatt acgacgtcga tgccattgta ccccaatcct ttttgaagga
     4381 cgattcaatc gacaataaag tgcttacacg ctcggataag aaccgaggga aaagtgacaa
     4441 tgttccaagc gaggaagtcg taaagaaaat gaagaactat tggcggcagc tcctaaatgc
     4501 gaaactgata acgcaaagaa agttcgataa cttaactaaa gctgagaggg gtggcttgtc
     4561 tgaacttgac aaggccggat ttattaaacg tcagctcgtg gaaacccgcc aaatcacaaa
     4621 gcatgttgca cagatactag attcccgaat gaatacgaaa tacgacgaga acgataagct
     4681 gattcgggaa gtcaaagtaa tcactttaaa gtcaaaattg gtgtcggact tcagaaagga
     4741 ttttcaattc tataaagtta gggagataaa taactaccac catgcgcacg acgcttatct
     4801 taatgccgtc gtagggaccg cactcattaa gaaatacccg aagctagaaa gtgagtttgt
     4861 gtatggtgat tacaaagttt atgacgtccg taagatgatc gcgaaaagcg aacaggagat
     4921 aggcaaggct acagccaaat acttctttta ttctaacatt atgaatttct ttaagacgga
     4981 aatcactctg gcaaacggag agatacgcaa acgaccttta attgaaacca atggggagac
     5041 aggtgaaatc gtatgggata agggccggga cttcgcgacg gtgagaaaag ttttgtccat
     5101 gccccaagtc aacatagtaa agaaaactga ggtgcagacc ggagggtttt caaaggaatc
     5161 gattcttcca aaaaggaata gtgataagct catcgctcgt aaaaaggact gggacccgaa
     5221 aaagtacggt ggcttcgata gccctacagt tgcctattct gtcctagtag tggcaaaagt
     5281 tgagaaggga aaatccaaga aactgaagtc agtcaaagaa ttattgggga taacgattat
     5341 ggagcgctcg tcttttgaaa agaaccccat cgacttcctt gaggcgaaag gttacaagga
     5401 agtaaaaaag gatctcataa ttaaactacc aaagtatagt ctgtttgagt tagaaaatgg
     5461 ccgaaaacgg atgttggcta gcgccggaga gcttcaaaag gggaacgaac tcgcactacc
     5521 gtctaaatac gtgaatttcc tgtatttagc gtcccattac gagaagttga aaggttcacc
     5581 tgaagataac gaacagaagc aactttttgt tgagcagcac aaacattatc tcgacgaaat
     5641 catagagcaa atttcggaat tcagtaagag agtcatccta gctgatgcca atctggacaa
     5701 agtattaagc gcatacaaca agcacaggga taaacccata cgtgagcagg cggaaaatat
     5761 tatccatttg tttactctta ccaacctcgg cgctccagcc gcattcaagt attttgacac
     5821 aacgatagat cgcaaacgat acacttctac caaggaggtg ctagacgcga cactgattca
     5881 ccaatccatc acgggattat atgaaactcg gatagatttg tcacagcttg ggggtgacgg
     5941 atcccccaag aagaagagga aagtcaaaag gccggcggcc acgaaaaagg ccggccaggc
     6001 aaaaaagaaa aagggatcct acccatacga tgttccagat tacgcttatc cctacgacgt
     6061 gcctgattat gcatacccat atgatgtccc cgactatgcc ggaagcgagg ccagcggttc
     6121 cggacgggct gacgcattgg acgattttga tctggatatg ctgggaagtg acgccctcga
     6181 tgattttgac cttgacatgc ttggttcgga tgcccttgat gactttgacc tcgacatgct
     6241 cggcagtgac gcccttgatg atttcgacct ggacatgctg attaactcta gaagttccgg
     6301 atctccgaaa aagaaacgca aagttggtag ccagtacctg cccgacaccg acgaccggca
     6361 ccggatcgag gaaaagcgga agcggaccta cgagacattc aagagcatca tgaagaagtc
     6421 ccccttcagc ggccccaccg accctagacc tccacctaga agaatcgccg tgcccagcag
     6481 atccagcgcc agcgtgccaa aacctgcccc ccagccttac cccttcacca gcagcctgag
     6541 caccatcaac tacgacgagt tccctaccat ggtgttcccc agcggccaga tctctcaggc
     6601 ctctgctctg gctccagccc ctcctcaggt gctgcctcag gctcctgctc ctgcaccagc
     6661 tccagccatg gtgtctgcac tggctcaggc accagcaccc gtgcctgtgc tggctcctgg
     6721 acctccacag gctgtggctc caccagcccc taaacctaca caggccggcg agggcacact
     6781 gtctgaagct ctgctgcagc tgcagttcga cgacgaggat ctgggagccc tgctgggaaa
     6841 cagcaccgat cctgccgtgt tcaccgacct ggccagcgtg gacaacagcg agttccagca
     6901 gctgctgaac cagggcatcc ctgtggcccc tcacaccacc gagcccatgc tgatggaata
     6961 ccccgaggcc atcacccggc tcgtgacagg cgctcagagg cctcctgatc cagctcctgc
     7021 ccctctggga gcaccaggcc tgcctaatgg actgctgtct ggcgacgagg acttcagctc
     7081 tatcgccgat atggatttct cagccttgct gggctctggc agcggcagcc gggattccag
     7141 ggaagggatg tttttgccga agcctgaggc cggctccgct attagtgacg tgtttgaggg
     7201 ccgcgaggtg tgccagccaa aacgaatccg gccatttcat cctccaggaa gtccatgggc
     7261 caaccgccca ctccccgcca gcctcgcacc aacaccaacc ggtccagtac atgagccagt
     7321 cgggtcactg accccggcac cagtccctca gccactggat ccagcgcccg cagtgactcc
     7381 cgaggccagt cacctgttgg aggatcccga tgaagagacg agccaggctg tcaaagccct
     7441 tcgggagatg gccgatactg tgattcccca gaaggaagag gctgcaatct gtggccaaat
     7501 ggacctttcc catccgcccc caaggggcca tctggatgag ctgacaacca cacttgagtc
     7561 catgaccgag gatctgaacc tggactcacc cctgaccccg gaattgaacg agattctgga
     7621 taccttcctg aacgacgagt gcctcttgca tgccatgcat atcagcacag gactgtccat
     7681 cttcgacaca tctctgtttt aaggccgcac tcctcaggtg caggctgcct atcagaaggt
     7741 ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc
     7801 caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa
     7861 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata
     7921 tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata
     7981 tgcccatatg ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa
     8041 acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt
     8101 ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt
     8161 tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct
     8221 cttatggaga tccctcgacc tgcagcccaa gcttggcgta atcatggtca tagctgtttc
     8281 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     8341 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     8401 ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt agtcagcaac
     8461 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     8521 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     8581 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     8641 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     8701 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     8761 tatcatgtct ggatccgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     8821 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     8881 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     8941 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     9001 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     9061 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     9121 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     9181 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     9241 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     9301 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     9361 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     9421 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     9481 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     9541 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     9601 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     9661 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     9721 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     9781 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     9841 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     9901 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     9961 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
    10021 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
    10081 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
    10141 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
    10201 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
    10261 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
    10321 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
    10381 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
    10441 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
    10501 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
    10561 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
    10621 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
    10681 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
    10741 catttccccg aaaagtgcca cctgggtc
//
