LOCUS       sequence-76544-e       12794 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive dCas9-VP64 lentiviral expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12794)
  AUTHORS   Kearns NA, Genga RM, Enuameh MS, Garber M, Wolfe SA, Maehr R
  TITLE     Cas9 effector-mediated regulation of transcription and 
            differentiation in human pluripotent stem cells.
  JOURNAL   Development. 2014 Jan;141(1):219-23. doi: 10.1242/dev.103341.
  PUBMED    24346702
REFERENCE   2  (bases 1 to 12794)
  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: "10.1242/dev"; 
            journalName: "Development"; date: "2014-01"; volume: "141"; issue: 
            "1"; pages: "219-23"
COMMENT     Sequence Label: pHAGE EF1-alpha dCas9-VP64
FEATURES             Location/Qualifiers
     source          1..12794
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             67..93
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        126..625
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(161..184)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     558..577
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             646..1245
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(646..665)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1142..1162
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    1262..1850
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1315..1335)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1733..1744)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1925..2158
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(2277..2295)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2363..2467
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2468..2536,2537..3328)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2686..2705)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3499..4087
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3988..4007
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4241..4258
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4498..4519
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4534..4539,4540..4557,4558..4564)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4572..4588
                     /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)."
     primer_bind     4577..4599
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4596..4612
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4596..4612
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4662..4681
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             4873..5506
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    5553..5678
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6175..6408
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6593..6637
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    6930..7047
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        7093..8271
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          7324..8262
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     8219..8239
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    8306..8330
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      8334..8343
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(8340..8366,8367..8369,8370..10853,10854..10859,
                     10860..10925,10926..10928,10929..12443)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=Cas9m4
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D10A, D839A, 
                     H840A, and N863A (Mali et al., 2013)"
                     /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             join(8340..8366,8367..8369,8370..10856,10857..10859,
                     10860..12443)
                     /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             8340..12443
                     /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="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             12456..12476
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             12477..12566
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             12588..12737
                     /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"
     protein_bind    complement(join(12784..12794,1..14))
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 ttgtacaaag tggttgatat ccagcacagt ggcggccgct cgagtctaga gggcccgcgg
       61 ttcgaatacc catacgacgt cccagactac gcttagtaat gattaattaa actagaaatt
      121 ctaccgggta ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt tagcagcccc
      181 gctgggcact tggcgctaca caagtggcct ctggcctcgc acacattcca catccaccgg
      241 taggcgccaa ccggctccgt tctttggtgg ccccttcgcg ccaccttcta ctcctcccct
      301 agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt
      361 agcacgtctc actagtctcg tgcagatgga cagcaccgct gagcaatgga agcgggtagg
      421 cctttggggc agcggccaat agcagctttg ctccttcgct ttctgggctc agaggctggg
      481 aaggggtggg tccgggggcg ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag
      541 gtcctccgga ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc
      601 ctcttcctca tctccgggcc tttcgacctg cagcccaagc ttaccatgac cgagtacaag
      661 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
      721 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
      781 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
      841 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
      901 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
      961 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     1021 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     1081 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     1141 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     1201 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaggtac cgatagatcc
     1261 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     1321 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     1381 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     1441 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     1501 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     1561 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     1621 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     1681 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     1741 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     1801 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctgagatcct
     1861 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg
     1921 ggactggaag ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg
     1981 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
     2041 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
     2101 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
     2161 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag
     2221 agtgagaggc ccgggttaat taaggaaagg gctagatcat tcttgaagac gaaagggcct
     2281 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     2341 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     2401 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     2461 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     2521 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     2581 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     2641 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     2701 attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     2761 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     2821 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     2881 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     2941 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     3001 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     3061 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     3121 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     3181 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     3241 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     3301 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     3361 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     3421 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     3481 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3541 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3601 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     3661 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3721 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3781 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3841 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3901 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3961 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4021 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4081 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4141 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     4201 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     4261 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     4321 cagcaagctc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     4381 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     4441 ctccccgtgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     4501 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     4561 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg acatgattac
     4621 gaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4681 caatgtatct tatcatgtct ggatcaactg gataactcaa gctaaccaaa atcatcccaa
     4741 acttcccacc ccatacccta ttaccactgc caattacctg tggtttcatt tactctaaac
     4801 ctgtgattcc tctgaattat tttcatttta aagaaattgt atttgttaaa tatgtactac
     4861 aaacttagta gttggaaggg ctaattcact cccaaagaag acaagatatc cttgatctgt
     4921 ggatctacca cacacaaggc tacttccctg attagcagaa ctacacacca gggccagggg
     4981 tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag ccagataagg
     5041 tagaagaggc caataaagga gagaacacca gcttgttaca ccctgtgagc ctgcatggga
     5101 tggatgaccc ggagagagaa gtgttagagt ggaggtttga cagccgccta gcatttcatc
     5161 acgtggcccg agagctgcat ccggagtact tcaagaactg ctgatatcga gcttgctaca
     5221 agggactttc cgctggggac tttccaggga ggcgtggcct gggcgggact ggggagtggc
     5281 gagccctcag atcctgcata taagcagctg ctttttgcct gtactgggtc tctctggtta
     5341 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     5401 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     5461 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca
     5521 gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg
     5581 aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta
     5641 gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta
     5701 gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac
     5761 atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa
     5821 catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag
     5881 aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag
     5941 agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga
     6001 ccaccgcaca gcaagcggcc ggccgctgat cttcagacct ggaggaggag atatgaggga
     6061 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     6121 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     6181 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     6241 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     6301 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     6361 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     6421 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     6481 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
     6541 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     6601 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     6661 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     6721 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     6781 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     6841 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcgccg
     6901 aattcacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat tggggggtac
     6961 agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa
     7021 aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag agatccagtt
     7081 tggactagcg atggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc
     7141 gagaagttgt ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta
     7201 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg
     7261 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca
     7321 caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc
     7381 gtgccttgaa ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt
     7441 ggaagtgggt gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga
     7501 gttgaggcct ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc
     7561 tgtctcgctg ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg
     7621 ctttttttct ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg
     7681 gtttttgggg ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg
     7741 cggggcctgc gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct
     7801 gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc
     7861 cggtcggcac cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc
     7921 tcaaaatgga ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa
     7981 agggcctttc cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc
     8041 aggcacctcg attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg
     8101 ttttatgcga tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg
     8161 cacttgatgt aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc
     8221 aagcctcaga cagtggttca aagttttttt cttccatttc aggtgtcgtg aactagtcca
     8281 gtgtggtgga attctgcaga tatcaacaag tttgtacaaa aaagcaggct ggcgccacca
     8341 tggacaagaa gtactccatt gggctcgcta tcggtaccaa cagcgtcggc tgggccgtca
     8401 ttacggacga gtacaaggtg ccgagcaaaa aattcaaagt tctgggcaat accgatcgcc
     8461 acagcataaa gaagaacctc attggagccc tcctgttcga ctccggggag acggccgaag
     8521 ccacgcggct caaaagaaca gcacggcgca gatatacccg cagaaagaat cggatctgct
     8581 acctgcagga gatctttagt aatgagatgg ctaaggtgga tgactctttc ttccataggc
     8641 tggaggagtc ctttttggtg gaggaggata aaaagcacga gcgccaccca atctttggca
     8701 atatcgtgga cgaggtggcg taccatgaaa agtacccaac catatatcat ctgaggaaga
     8761 agctggtaga cagtactgat aaggctgact tgcggttgat ctatctcgcg ctggcgcaca
     8821 tgatcaaatt tcggggacac ttcctcatcg agggggacct gaacccagac aacagcgatg
     8881 tcgacaaact ctttatccaa ctggttcaga cttacaatca gcttttcgag gagaacccga
     8941 tcaacgcatc cggcgttgac gccaaagcaa tcctgagcgc taggctgtcc aaatcccggc
     9001 ggctcgaaaa cctcatcgca cagctccctg gggagaagaa gaacggcctg tttggtaatc
     9061 ttatcgccct gtcactcggg ctgaccccca actttaaatc taacttcgac ctggccgaag
     9121 atgccaagct gcaactgagc aaagacacct acgatgatga tctcgacaat ctgctggccc
     9181 agatcggcga ccagtacgca gacctttttt tggcggcaaa gaacctgtca gacgccattc
     9241 tgctgagtga tattctgcga gtgaacacgg agatcaccaa agctccgctg agcgctagta
     9301 tgatcaagcg ctatgatgag caccaccaag acttgacttt gctgaaggcc cttgtcagac
     9361 agcaactgcc tgagaagtac aaggaaattt tcttcgatca gtctaaaaat ggctacgccg
     9421 gatacattga cggcggagca agccaggagg aattttacaa atttattaag cccatcttgg
     9481 aaaaaatgga cggcaccgag gagctgctgg taaagctgaa cagagaagat ctgttgcgca
     9541 aacagcgcac tttcgacaat ggaagcatcc cccaccagat tcacctgggc gaactgcacg
     9601 ctatcctcag gcggcaagag gatttctacc cctttttgaa agataacagg gaaaagattg
     9661 agaaaatcct cacatttcgg ataccctact atgtaggccc cctcgctcgg ggaaattcca
     9721 gattcgcgtg gatgactcgc aaatcagaag agaccatcac tccctggaac ttcgaggaag
     9781 tcgtggataa gggggcctct gcccagtcct tcatcgaaag gatgactaac tttgataaaa
     9841 atctgcctaa cgaaaaggtg cttcctaaac actctctgct gtacgagtac ttcacagttt
     9901 ataacgagct caccaaggtc aaatacgtca cagaagggat gagaaagcca gcattcctgt
     9961 ctggagagca gaagaaagct atcgtggacc tcctcttcaa gacgaaccgg aaagttaccg
    10021 tgaaacagct caaagaagac tatttcaaaa agattgaatg tttcgactct gttgaaatca
    10081 gcggagtgga ggatcgcttc aacgcatccc tgggaacgta tcacgatctc ctgaaaatca
    10141 ttaaagacaa ggacttcctg gacaatgagg agaacgagga cattcttgag gacattgtcc
    10201 tcacccttac gttgtttgaa gatagggaga tgattgaaga acgcttgaaa acttacgctc
    10261 atctcttcga cgacaaagtc atgaaacagc tcaagagacg ccgatataca ggatgggggc
    10321 ggctgtcaag aaaactgatc aatggcatcc gagacaagca gagtggaaag acaatcctgg
    10381 attttcttaa gtccgatgga tttgccaacc ggaacttcat gcagttgatc catgatgact
    10441 ctctcacctt taaggaggac atccagaaag cacaagtttc tggccagggg gacagtcttc
    10501 acgagcacat cgctaatctt gcaggtagcc cagctatcaa aaagggaata ctgcagaccg
    10561 ttaaggtcgt ggatgaactc gtcaaagtaa tgggaaggca taagcccgag aatatcgtta
    10621 tcgagatggc ccgagagaac caaactaccc agaagggaca gaagaacagt agggaaagga
    10681 tgaagaggat tgaagagggt ataaaagaac tggggtccca aatccttaag gaacacccag
    10741 ttgaaaacac ccagcttcag aatgagaagc tctacctgta ctacctgcag aacggcaggg
    10801 acatgtacgt ggatcaggaa ctggacatca accggttgtc cgactacgac gtggatgcta
    10861 tcgtgcccca aagctttctc aaagatgatt ctattgataa taaagtgttg acaagatccg
    10921 ataaaaatag agggaagagt gataacgtcc cctcagaaga agttgtcaag aaaatgaaaa
    10981 attattggcg gcagctgctg aacgccaaac tgatcacaca acggaagttc gataatctga
    11041 ctaaggctga acgaggtggc ctgtctgagt tggataaagc cggcttcatc aaaaggcagc
    11101 ttgttgagac acgccagatc accaagcacg tggcccaaat tctcgattca cgcatgaaca
    11161 ccaagtacga tgaaaatgac aaactgattc gagaggtgaa agttattact ctgaagtcta
    11221 agctggtctc agatttcaga aaggactttc agttttataa ggtgagagag atcaacaatt
    11281 accaccatgc gcatgatgcc tacctgaatg cagtggtagg cactgcactt atcaaaaaat
    11341 atcccaagct ggaatctgaa tttgtttacg gagactataa agtgtacgat gttaggaaaa
    11401 tgatcgcaaa gtctgagcag gaaataggca aggccaccgc taagtacttc ttttacagca
    11461 atattatgaa ttttttcaag accgagatta cactggccaa tggagagatt cggaagcgac
    11521 cacttatcga aacaaacgga gaaacaggag aaatcgtgtg ggacaagggt agggatttcg
    11581 cgacagtccg caaggtcctg tccatgccgc aggtgaacat cgttaaaaag accgaagtac
    11641 agaccggagg cttctccaag gaaagtatcc tcccgaaaag gaacagcgac aagctgatcg
    11701 cacgcaaaaa agattgggac cccaagaaat acggcggatt cgattctcct acagtcgctt
    11761 acagtgtact ggttgtggcc aaagtggaga aagggaagtc taaaaaactc aaaagcgtca
    11821 aggaactgct gggcatcaca atcatggagc gatccagctt cgagaaaaac cccatcgact
    11881 ttctcgaagc gaaaggatat aaagaggtca aaaaagacct catcattaag ctgcccaagt
    11941 actctctctt tgagcttgaa aacggccgga aacgaatgct cgctagtgcg ggcgagctgc
    12001 agaaaggtaa cgagctggca ctgccctcta aatacgttaa tttcttgtat ctggccagcc
    12061 actatgaaaa gctcaaaggg tctcccgaag ataatgagca gaagcagctg ttcgtggaac
    12121 aacacaaaca ctaccttgat gagatcatcg agcaaataag cgagttctcc aaaagagtga
    12181 tcctcgccga cgctaacctc gataaggtgc tttctgctta caataagcac agggataagc
    12241 ccatcaggga gcaggcagaa aacattatcc acttgtttac tctgaccaac ttgggcgcgc
    12301 ctgcagcctt caagtacttc gacaccacca tagacagaaa gcggtacacc tctacaaagg
    12361 aggtcctgga cgccacactg attcatcagt caattacggg gctctatgaa acaagaatcg
    12421 acctctctca gctcggtgga gacggcaccg gcgggcccaa gaagaagagg aaggtatacc
    12481 catacgatgt tcctgactat gcgggctatc cctatgacgt cccggactat gcaggatcgt
    12541 atccttatga cgttccagat tacgctggat ccggttccgg acgggctgac gcattggacg
    12601 attttgatct ggatatgctg ggaagtgacg ccctcgatga ttttgacctt gacatgcttg
    12661 gttcggatgc ccttgatgac tttgacctcg acatgctcgg cagtgacgcc cttgatgatt
    12721 tcgacctgga catgctgatt aactaattcg aatccggata agcggccgcc tcgagatatc
    12781 tagacccagc tttc
//
