LOCUS       sequence-160890-       13189 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses direct fusion VPR-Sp dCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13189)
  AUTHORS   Gao Y, Xiong X, Wong S, Charles EJ, Lim WA, Qi LS
  TITLE     Complex transcriptional modulation with orthogonal and inducible 
            dCas9 regulators.
  JOURNAL   Nat Methods. 2016 Oct 24. doi: 10.1038/nmeth.4042.
  PUBMED    27776111
REFERENCE   2  (bases 1 to 13189)
  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. 2016 Oct 24. doi: 10.1038/nmeth.4042."
COMMENT     Sequence Label: pSLQ2814 pPB: CAG-Puro-WPRE PGK-VPR-tagBFP-Spd...
FEATURES             Location/Qualifiers
     source          1..13189
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..284
                     /label=chicken beta-actin promoter
     intron          286..1303
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1227..1249
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1311..1330
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1370..1969
                     /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(1370..1389)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1866..1886
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    2007..2595
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2060..2080)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2478..2489)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        2616..3115
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2651..2674)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3048..3067
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      3139..3148
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3148..3297
                     /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             3304..3324
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3754..4125
                     /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
                     EDFSSIADMDFSALLSQISS"
     CDS             4144..4713
                     /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"
     CDS             4741..5439
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     regulatory      5455..5464
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5461..5487,5488..5490,5491..7977,7978..7980,
                     7981..9564)
                     /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             5461..9564
                     /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             9568..9594
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             9574..9600
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             9613..9633
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9640..9660
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     repeat_region   9877..9939
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(9990..10010)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(9990..10008)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(10029..10045)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10029..10045)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10042..10064)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10053..10069
                     /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(10077..10083,10084..10101,10102..10107))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10122..10143
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10260..10277)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10431..11019)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10511..10530)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11190..11981,11982..12050))
                     /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     11813..11832
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12051..12155)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(12181..12636)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12318..12339)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     12530..12549
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12763..12785
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     12777..12794
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     12778..12794
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12804..12823
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        12804..12822
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   12852..12886
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac
       61 ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg
      121 ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga
      181 gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc
      241 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcggggagt cgctgcgacg
      301 ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact
      361 gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta
      421 gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct
      481 ccgggagggc cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt
      541 ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc
      601 ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg
      661 gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg tgggggggtg
      721 agcagggggt gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc ctccccgagt
      781 tgctgagcac ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg cggggctcgc
      841 cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc cgcctcgggc
      901 cggggagggc tcgggggagg ggcgcggcgg cccccggagc gccggcggct gtcgaggcgc
      961 ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg gacttccttt
     1021 gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct ctagcgggcg
     1081 cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc
     1141 gccgcgccgc cgtccccttc tccctctcca gcctcggggc tgtccgcggg gggacggctg
     1201 ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg gcggctctag
     1261 agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct
     1321 ggttattgtg ctgtctcatc attttggcaa agaattcgcc accatgacca tgaccgagta
     1381 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc
     1441 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga
     1501 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
     1561 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
     1621 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
     1681 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
     1741 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
     1801 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
     1861 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
     1921 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgag tttaaaccat
     1981 gcaagcttga tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat
     2041 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     2101 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     2161 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     2221 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     2281 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     2341 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     2401 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     2461 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     2521 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     2581 gggccgcctc cccgcatcct gcaggtaatt ctaccgggta ggggaggcgc ttttcccaag
     2641 gcagtctgga gcatgcgctt tagcagcccc gctgggcact tggcgctaca caagtggcct
     2701 ctggcctcgc acacattcca catccaccgg taggcgccaa ccggctccgt tctttggtgg
     2761 ccccttcgcg ccaccttcta ctcctcccct agtcaggaag ttcccccccg ccccgcagct
     2821 cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc actaggctcg tgcagatgga
     2881 cagcaccgct gagcaatgga agcgggtagg cctttggggc agcggccaat agcagctttg
     2941 ctccttcgct ttctgggctc agaggctggg aaggggtggg tccgggggcg ggctcagggg
     3001 cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga ggcccggcat tctgcacgct
     3061 tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca tctccgggcc tttcgacctg
     3121 cagcccaagc ttgtcgacgc caccatggat gctttagacg attttgactt agatatgctt
     3181 ggttcagacg cgttagacga cttcgaccta gacatgttag gctcagatgc attggacgac
     3241 ttcgatttag atatgttggg ctccgatgcc ctagatgact ttgatctaga tatgctaggt
     3301 agtcccaaaa agaagaggaa agtgggatcc cagtatctgc ccgacacaga tgatagacac
     3361 cgaatcgaag agaaacgcaa gcgaacgtat gaaaccttca aatcgatcat gaagaaatcg
     3421 cccttctcgg gtccgaccga tcccaggccc ccaccgagaa ggattgcggt cccgtcccgc
     3481 tcgtcggcca gcgtgccgaa gcctgcgccg cagccctacc ccttcacgtc gagcctgagc
     3541 acaatcaatt atgacgagtt cccgacgatg gtgttcccct cgggacaaat ctcacaagcc
     3601 tcggcgctcg caccagcgcc tccccaagtc cttccgcaag cgcctgcccc agcgcctgca
     3661 ccggcaatgg tgtccgccct cgcacaggcc cctgcgcccg tccccgtgct cgcgcctgga
     3721 ccgccccagg cggtcgctcc accggctccg aagccgacgc aggccggaga gggaacactc
     3781 tccgaagcac ttcttcaact ccagtttgat gacgaggatc ttggagcact ccttggaaac
     3841 tcgacagacc ctgcggtgtt taccgacctc gcgtcagtag ataactccga atttcagcag
     3901 cttttgaacc agggtatccc ggtcgcgcca catacaacgg agcccatgtt gatggaatac
     3961 cccgaagcaa tcacgagact tgtgacggga gcgcagcggc ctcccgatcc cgcacccgca
     4021 cctttggggg cacctggcct ccctaacgga cttttgagcg gcgacgagga tttctcctcc
     4081 atcgccgata tggatttctc agccttgctg tcacagattt ccagcggctc tggcagcggc
     4141 agccgggatt ccagggaagg gatgtttttg ccgaagcctg aggccggctc cgctattagt
     4201 gacgtgtttg agggccgcga ggtgtgccag ccaaaacgaa tccggccatt tcatcctcca
     4261 ggaagtccat gggccaaccg cccactcccc gccagcctcg caccaacacc aaccggtcca
     4321 gtacatgagc cagtcgggtc actgaccccg gcaccagtcc ctcagccact ggatccagcg
     4381 cccgcagtga ctcccgaggc cagtcacctg ttggaggatc ccgatgaaga gacgagccag
     4441 gctgtcaaag cccttcggga gatggccgat actgtgattc cccagaagga agaggctgca
     4501 atctgtggcc aaatggacct ttcccatccg cccccaaggg gccatctgga tgagctgaca
     4561 accacacttg agtccatgac cgaggatctg aacctggact cacccctgac cccggaattg
     4621 aacgagattc tggatacctt cctgaacgac gagtgcctct tgcatgccat gcatatcagc
     4681 acaggactgt ccatcttcga cacatctctg tttcatatgt caagcgctgc aggtgctacc
     4741 atgagcgagc tgattaagga gaacatgcac atgaagctgt acatggaggg caccgtggac
     4801 aaccatcact tcaagtgcac atccgagggc gaaggcaagc cctacgaggg cacccagacc
     4861 atgagaatca aggtggtcga gggcggccct ctccccttcg ccttcgacat cctggctact
     4921 agcttcctct acggcagcaa gaccttcatc aaccacaccc agggcatccc cgacttcttc
     4981 aagcagtcct tccctgaggg cttcacatgg gagagagtca ccacatacga agacgggggc
     5041 gtgctgaccg ctacccagga caccagcctc caggacggct gcctcatcta caacgtcaag
     5101 atcagagggg tgaacttcac atccaacggc cctgtgatgc agaagaaaac actcggctgg
     5161 gaggccttca ccgagacgct gtaccccgct gacggcggcc tggaaggcag aaacgacatg
     5221 gccctgaagc tcgtgggcgg gagccatctg atcgcaaaca tcaagaccac atatagatcc
     5281 aagaaacccg ctaagaacct caagatgcct ggcgtctact atgtggacta cagactggaa
     5341 agaatcaagg aggccaacaa cgagacctac gtcgagcagc acgaggtggc agtggccaga
     5401 tactgcgacc tccctagcaa actggggcac aagcttaatt caagcgctgc aggtgccacc
     5461 atggacaaga agtacagcat cggcctggcc atcggcacca actctgtggg ctgggccgtg
     5521 atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa caccgaccgg
     5581 cacagcatca agaagaacct gatcggcgcc ctgctgttcg acagcggaga aacagccgag
     5641 gccacccggc tgaagagaac cgccagaaga agatacacca gacggaagaa ccggatctgc
     5701 tatctgcaag agatcttcag caacgagatg gccaaggtgg acgacagctt cttccacaga
     5761 ctggaagagt ccttcctggt ggaagaggat aagaagcacg agcggcaccc catcttcggc
     5821 aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca cctgagaaag
     5881 aaactggtgg acagcaccga caaggccgac ctgcggctga tctatctggc cctggcccac
     5941 atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac
     6001 gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga ggaaaacccc
     6061 atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg ccagactgag caagagcaga
     6121 cggctggaaa atctgatcgc ccagctgccc ggcgagaaga agaatggcct gttcggcaac
     6181 ctgattgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag
     6241 gatgccaaac tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc
     6301 cagatcggcg accagtacgc cgacctgttt ctggccgcca agaacctgtc cgacgccatc
     6361 ctgctgagcg acatcctgag agtgaacacc gagatcacca aggcccccct gagcgcctct
     6421 atgatcaaga gatacgacga gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg
     6481 cagcagctgc ctgagaagta caaagagatt ttcttcgacc agagcaagaa cggctacgcc
     6541 ggctacatcg atggcggagc cagccaggaa gagttctaca agttcatcaa gcccatcctg
     6601 gaaaagatgg acggcaccga ggaactgctc gtgaagctga acagagagga cctgctgcgg
     6661 aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg agagctgcac
     6721 gccattctgc ggcggcagga agatttttac ccattcctga aggacaaccg ggaaaagatc
     6781 gagaagatcc tgaccttccg catcccctac tacgtgggcc ctctggccag gggaaacagc
     6841 agattcgcct ggatgaccag aaagagcgag gaaaccatca ccccctggaa cttcgaggaa
     6901 gtggtggaca agggcgccag cgcccagagc ttcatcgagc ggatgaccaa cttcgataag
     6961 aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg
     7021 tacaacgagc tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc cgccttcctg
     7081 agcggcgagc agaaaaaagc catcgtggac ctgctgttca agaccaaccg gaaagtgacc
     7141 gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc
     7201 tccggcgtgg aagatcggtt caacgcctcc ctgggcacat accacgatct gctgaaaatt
     7261 atcaaggaca aggacttcct ggacaatgag gaaaacgagg acattctgga agatatcgtg
     7321 ctgaccctga cactgtttga ggacagagag atgatcgagg aacggctgaa aacctatgcc
     7381 cacctgttcg acgacaaagt gatgaagcag ctgaagcggc ggagatacac cggctggggc
     7441 aggctgagcc ggaagctgat caacggcatc cgggacaagc agtccggcaa gacaatcctg
     7501 gatttcctga agtccgacgg cttcgccaac agaaacttca tgcagctgat ccacgacgac
     7561 agcctgacct ttaaagagga catccagaaa gcccaggtgt ccggccaggg cgatagcctg
     7621 cacgagcaca ttgccaatct ggccggcagc cccgccatta agaagggcat cctgcagaca
     7681 gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc acaagcccga gaacatcgtg
     7741 atcgaaatgg ccagagagaa ccagaccacc cagaagggac agaagaacag ccgcgagaga
     7801 atgaagcgga tcgaagaggg catcaaagag ctgggcagcc agatcctgaa agaacacccc
     7861 gtggaaaaca cccagctgca gaacgagaag ctgtacctgt actacctgca gaatgggcgg
     7921 gatatgtacg tggaccagga actggacatc aaccggctgt ccgactacga tgtggacgct
     7981 atcgtgcctc agagctttct gaaggacgac tccatcgata acaaagtgct gactcggagc
     8041 gacaagaacc ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag
     8101 aactactggc gccagctgct gaatgccaag ctgattaccc agaggaagtt cgacaatctg
     8161 accaaggccg agagaggcgg cctgagcgaa ctggataagg ccggcttcat caagagacag
     8221 ctggtggaaa cccggcagat cacaaagcac gtggcacaga tcctggactc ccggatgaac
     8281 actaagtacg acgagaacga caaactgatc cgggaagtga aagtgatcac cctgaagtcc
     8341 aagctggtgt ccgatttccg gaaggatttc cagttttaca aagtgcgcga gatcaacaac
     8401 taccaccacg cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag
     8461 taccctaagc tggaaagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag
     8521 atgatcgcca agagcgagca ggaaatcggc aaggctaccg ccaagtactt cttctacagc
     8581 aacatcatga actttttcaa gaccgagatt accctggcca acggcgagat ccggaagcgg
     8641 cctctgatcg agacaaacgg cgaaacaggc gagatcgtgt gggataaggg ccgggacttt
     8701 gccaccgtgc ggaaagtgct gtctatgccc caagtgaata tcgtgaaaaa gaccgaggtg
     8761 cagacaggcg gcttcagcaa agagtctatc ctgcccaaga ggaacagcga caagctgatc
     8821 gccagaaaga aggactggga ccctaagaag tacggcggct tcgacagccc caccgtggcc
     8881 tattctgtgc tggtggtggc caaagtggaa aagggcaagt ccaagaaact gaagagtgtg
     8941 aaagagctgc tggggatcac catcatggaa agaagcagct tcgagaagaa tcccatcgac
     9001 tttctggaag ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag
     9061 tactccctgt tcgagctgga aaacggccgg aagagaatgc tggcctctgc cggcgaactg
     9121 cagaagggaa acgaactggc cctgccctcc aaatatgtga acttcctgta cctggccagc
     9181 cactatgaga agctgaaggg ctcccccgag gataatgagc agaaacagct gtttgtggaa
     9241 cagcacaaac actacctgga cgagatcatc gagcagatca gcgagttctc caagagagtg
     9301 atcctggccg acgctaatct ggacaaggtg ctgagcgcct acaacaagca cagagacaag
     9361 cctatcagag agcaggccga gaatatcatc cacctgttta ccctgaccaa tctgggagcc
     9421 cctgccgcct tcaagtactt tgacaccacc atcgaccgga agaggtacac cagcaccaaa
     9481 gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga gacacggatc
     9541 gacctgtctc agctgggagg cgacgcctat ccctatgacg tgcccgatta tgccagcctg
     9601 ggcagcggct cccccaagaa aaaacgcaag gtggaagatc ctaagaaaaa gcggaaagtg
     9661 gacggcattg gtagtgggag caacggatga ctcgagctag ttaaaagttt tgttacttta
     9721 tagaagaaat tttgagtttt tgtttttttt taataaataa ataaacataa ataaattgtt
     9781 tgttgaattt attattagta tgtaagtgta aatataataa aacttaatat ctattcaaat
     9841 taataaataa acctcgatat acagaccgat aaaacacatg cgtcaatttt acgcatgatt
     9901 atctttaacg tacgtcacaa tatgattatc tttctagggt taatctagta tacgcgtatg
     9961 cggccgctta attaatccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg
    10021 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
    10081 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
    10141 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
    10201 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
    10261 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
    10321 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
    10381 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
    10441 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
    10501 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
    10561 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
    10621 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
    10681 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
    10741 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
    10801 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
    10861 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
    10921 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
    10981 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
    11041 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
    11101 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
    11161 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
    11221 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
    11281 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
    11341 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
    11401 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
    11461 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
    11521 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
    11581 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
    11641 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
    11701 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
    11761 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
    11821 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
    11881 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
    11941 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
    12001 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
    12061 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
    12121 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
    12181 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
    12241 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
    12301 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
    12361 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
    12421 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
    12481 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
    12541 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
    12601 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca
    12661 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
    12721 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
    12781 aaacgacggc cagtgagcgc gcgtaatacg actcactata gggcgaattg gggcgcgcca
    12841 ttctagatta accctagaaa gatagtctgc gtaaaattga cgcatgcatt cttgaaatat
    12901 tgctctctct ttctaaatag cgcgaatccg tcgctgtgca tttaggacat ctcagtcgcc
    12961 gcttggagct cccgtgaggc gtgcttgtca atgcggtaag tgtcactgat tttgaactat
    13021 aacgaccgcg tgagtcaaaa tgacgcatga ttatctttta cgtgactttt aagatttaac
    13081 tcatacgata attatattgt tatttcatgt tctacttacg tgataactta ttatatatat
    13141 attttcttgt tatagatatc aactagaatg ctagcatggg cccagatct
//
