LOCUS       sequence-398036-        9815 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses inactivated S. pyogenes dCas9 (D10A, H840A) fused to VP64 
            transactivator domain in mammalian cells .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9815)
  AUTHORS   Perez-Pinera P, Kocak DD, Vockley CM, Adler AF, Kabadi AM, Polstein 
            LR, Thakore PI, Glass KA, Ousterout DG, Leong KW, Guilak F, Crawford
            GE, Reddy TE, Gersbach CA
  TITLE     RNA-guided gene activation by CRISPR-Cas9-based transcription 
            factors.
  JOURNAL   Nat Methods. 2013 Jul 25. doi: 10.1038/nmeth.2600.
  PUBMED    23892895
REFERENCE   2  (bases 1 to 9815)
  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"; journalName: "Nat 
            Methods. 2013 Jul 25. doi: 10.1038/nmeth.2600."
COMMENT     Sequence Label: pcDNA-dCas9-VP64
FEATURES             Location/Qualifiers
     source          1..9815
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(115..135)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        120..449
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      300..435
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     362..381
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             516..1310
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(570..589)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1180..1199
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1484..1605
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1521..1540)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1575..1594
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1654..1670)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1654..1670)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1667..1689)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1678..1694
                     /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(1702..1708,1709..1726,1727..1732))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1747..1768
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1885..1902)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2056..2644)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2136..2155)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2815..3606,3607..3675))
                     /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     3438..3457
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3676..3780)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3855..3874)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4046..4425
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4426..4629
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4580..4600
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4674..4693
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4674..4692
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4718..4783
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4790..4810
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(4820..4846,4847..4849,4850..7336,7337..7339,
                     7340..8923)
                     /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             8948..8968
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8978..9127
                     /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             9134..9160
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(9217..9234)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9223..9447
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(9493..9815,1..106)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9580..9599)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9790..9811
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
       61 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
      121 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
      181 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
      241 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
      301 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
      361 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
      421 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
      481 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
      541 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
      601 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
      661 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
      721 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
      781 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
      841 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
      901 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
      961 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     1021 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     1081 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     1141 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     1201 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     1261 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     1321 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     1381 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     1441 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     1501 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     1561 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     1621 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     1681 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     1741 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     1801 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     1861 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     1921 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     1981 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2041 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2101 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2161 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2221 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2281 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2341 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2401 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2461 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     2521 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2581 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2641 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     2701 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     2761 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     2821 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     2881 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     2941 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     3001 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     3061 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     3121 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     3181 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     3241 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     3301 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     3361 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     3421 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     3481 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     3541 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     3601 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     3661 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     3721 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     3781 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     3841 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     3901 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     3961 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc
     4021 gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca
     4081 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
     4141 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     4201 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     4261 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
     4321 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
     4381 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
     4441 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
     4501 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
     4561 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
     4621 agcagagctc tctggctaac tagagaaccc actgcttact ggcttatcga aattaatacg
     4681 actcactata gggagaccca agctggctag cgccatggac tacaaagacc atgacggtga
     4741 ttataaagat catgacatcg attacaagga tgacgatgac aagatggccc ccaagaagaa
     4801 gaggaaggtg ggccgcggaa tggataagaa atactcaata ggcttagcta tcggcacaaa
     4861 tagcgtcgga tgggcggtga tcactgatga atataaggtt ccgtctaaaa agttcaaggt
     4921 tctgggaaat acagaccgcc acagtatcaa aaaaaatctt ataggggctc ttttatttga
     4981 cagtggagag acagcggaag cgactcgtct caaacggaca gctcgtagaa ggtatacacg
     5041 tcggaagaat cgtatttgtt atctacagga gattttttca aatgagatgg cgaaagtaga
     5101 tgatagtttc tttcatcgac ttgaagagtc ttttttggtg gaagaagaca agaagcatga
     5161 acgtcatcct atttttggaa atatagtaga tgaagttgct tatcatgaga aatatccaac
     5221 tatctatcat ctgcgaaaaa aattggtaga ttctactgat aaagcggatt tgcgcttaat
     5281 ctatttggcc ttagcgcata tgattaagtt tcgtggtcat tttttgattg agggagattt
     5341 aaatcctgat aatagtgatg tggacaaact atttatccag ttggtacaaa cctacaatca
     5401 attatttgaa gaaaacccta ttaacgcaag tggagtagat gctaaagcga ttctttctgc
     5461 acgattgagt aaatcaagac gattagaaaa tctcattgct cagctccccg gtgagaagaa
     5521 aaatggctta tttgggaatc tcattgcttt gtcattgggt ttgaccccta attttaaatc
     5581 aaattttgat ttggcagaag atgctaaatt acagctttca aaagatactt acgatgatga
     5641 tttagataat ttattggcgc aaattggaga tcaatatgct gatttgtttt tggcagctaa
     5701 gaatttatca gatgctattt tactttcaga tatcctaaga gtaaatactg aaataactaa
     5761 ggctccccta tcagcttcaa tgattaaacg ctacgatgaa catcatcaag acttgactct
     5821 tttaaaagct ttagttcgac aacaacttcc agaaaagtat aaagaaatct tttttgatca
     5881 atcaaaaaac ggatatgcag gttatattga tgggggagct agccaagaag aattttataa
     5941 atttatcaaa ccaattttag aaaaaatgga tggtactgag gaattattgg tgaaactaaa
     6001 tcgtgaagat ttgctgcgca agcaacggac ctttgacaac ggctctattc cccatcaaat
     6061 tcacttgggt gagctgcatg ctattttgag aagacaagaa gacttttatc catttttaaa
     6121 agacaatcgt gagaagattg aaaaaatctt gacttttcga attccttatt atgttggtcc
     6181 attggcgcgt ggcaatagtc gttttgcatg gatgactcgg aagtctgaag aaacaattac
     6241 cccatggaat tttgaagaag ttgtcgataa aggtgcttca gctcaatcat ttattgaacg
     6301 catgacaaac tttgataaaa atcttccaaa tgaaaaagta ctaccaaaac atagtttgct
     6361 ttatgagtat tttacggttt ataacgaatt gacaaaggtc aaatatgtta ctgaaggaat
     6421 gcgaaaacca gcatttcttt caggtgaaca gaagaaagcc attgttgatt tactcttcaa
     6481 aacaaatcga aaagtaaccg ttaagcaatt aaaagaagat tatttcaaaa aaatagaatg
     6541 ttttgatagt gttgaaattt caggagttga agatagattt aatgcttcat taggtaccta
     6601 ccatgatttg ctaaaaatta ttaaagataa agattttttg gataatgaag aaaatgaaga
     6661 tatcttagag gatattgttt taacattgac cttatttgaa gatagggaga tgattgagga
     6721 aagacttaaa acatatgctc acctctttga tgataaggtg atgaaacagc ttaaacgtcg
     6781 ccgttatact ggttggggac gtttgtctcg aaaattgatt aatggtatta gggataagca
     6841 atctggcaaa acaatattag attttttgaa atcagatggt tttgccaatc gcaattttat
     6901 gcagctgatc catgatgata gtttgacatt taaagaagac attcaaaaag cacaagtgtc
     6961 tggacaaggc gatagtttac atgaacatat tgcaaattta gctggtagcc ctgctattaa
     7021 aaaaggtatt ttacagactg taaaagttgt tgatgaattg gtcaaagtaa tggggcggca
     7081 taagccagaa aatatcgtta ttgaaatggc acgtgaaaat cagacaactc aaaagggcca
     7141 gaaaaattcg cgagagcgta tgaaacgaat cgaagaaggt atcaaagaat taggaagtca
     7201 gattcttaaa gagcatcctg ttgaaaatac tcaattgcaa aatgaaaagc tctatctcta
     7261 ttatctccaa aatggaagag acatgtatgt ggaccaagaa ttagatatta atcgtttaag
     7321 tgattatgat gtcgatgcca ttgttccaca aagtttcctt aaagacgatt caatagacaa
     7381 taaggtctta acgcgttctg ataaaaatcg tggtaaatcg gataacgttc caagtgaaga
     7441 agtagtcaaa aagatgaaaa actattggag acaacttcta aacgccaagt taatcactca
     7501 acgtaagttt gataatttaa cgaaagctga acgtggaggt ttgagtgaac ttgataaagc
     7561 tggttttatc aaacgccaat tggttgaaac tcgccaaatc actaagcatg tggcacaaat
     7621 tttggatagt cgcatgaata ctaaatacga tgaaaatgat aaacttattc gagaggttaa
     7681 agtgattacc ttaaaatcta aattagtttc tgacttccga aaagatttcc aattctataa
     7741 agtacgtgag attaacaatt accatcatgc ccatgatgcg tatctaaatg ccgtcgttgg
     7801 aactgctttg attaagaaat atccaaaact tgaatcggag tttgtctatg gtgattataa
     7861 agtttatgat gttcgtaaaa tgattgctaa gtctgagcaa gaaataggca aagcaaccgc
     7921 aaaatatttc ttttactcta atatcatgaa cttcttcaaa acagaaatta cacttgcaaa
     7981 tggagagatt cgcaaacgcc ctctaatcga aactaatggg gaaactggag aaattgtctg
     8041 ggataaaggg cgagattttg ccacagtgcg caaagtattg tccatgcccc aagtcaatat
     8101 tgtcaagaaa acagaagtac agacaggcgg attctccaag gagtcaattt taccaaaaag
     8161 aaattcggac aagcttattg ctcgtaaaaa agactgggat ccaaaaaaat atggtggttt
     8221 tgatagtcca acggtagctt attcagtcct agtggttgct aaggtggaaa aagggaaatc
     8281 gaagaagtta aaatccgtta aagagttact agggatcaca attatggaaa gaagttcctt
     8341 tgaaaaaaat ccgattgact ttttagaagc taaaggatat aaggaagtta aaaaagactt
     8401 aatcattaaa ctacctaaat atagtctttt tgagttagaa aacggtcgta aacggatgct
     8461 ggctagtgcc ggagaattac aaaaaggaaa tgagctggct ctgccaagca aatatgtgaa
     8521 ttttttatat ttagctagtc attatgaaaa gttgaagggt agtccagaag ataacgaaca
     8581 aaaacaattg tttgtggagc agcataagca ttatttagat gagattattg agcaaatcag
     8641 tgaattttct aagcgtgtta ttttagcaga tgccaattta gataaagttc ttagtgcata
     8701 taacaaacat agagacaaac caatacgtga acaagcagaa aatattattc atttatttac
     8761 gttgacgaat cttggagctc ccgctgcttt taaatatttt gatacaacaa ttgatcgtaa
     8821 acgatatacg tctacaaaag aagttttaga tgccactctt atccatcaat ccatcactgg
     8881 tctttatgaa acacgcattg atttgagtca gctaggaggt gacccaattg ccggatccaa
     8941 ggctagcccg aaaaagaaac gcaaagttgg gcgcgccgac gcgctggacg atttcgatct
     9001 cgacatgctg ggttctgatg ccctcgatga ctttgacctg gatatgttgg gaagcgacgc
     9061 attggatgac tttgatctgg acatgctcgg ctccgatgct ctggacgatt tcgatctcga
     9121 tatgttaatt aactacccgt acgacgttcc ggactacgct tcttgaatcg gtaggaattc
     9181 gcggccgtct agacttaagt ttaaaccgct gatcagcctc gactgtgcct tctagttgcc
     9241 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     9301 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     9361 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     9421 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     9481 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     9541 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     9601 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     9661 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     9721 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     9781 tctttaatag tggactcttg ttccaaactg gaaca
//
