LOCUS       sequence-256768-       15275 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CAG-DNCR2-VPR-IRES-Puro-WPRE-SV40PA-PGK-NS3aH1-tagBFP-SpdCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15275)
  AUTHORS   Foight GW, Wang Z, Wei CT, Jr Greisen P, Warner KM, 
            Cunningham-Bryant D, Park K, Brunette TJ, Sheffler W, Baker D, Maly 
            DJ
  TITLE     Multi-input chemical control of protein dimerization for programming
            graded cellular responses.
  JOURNAL   Nat Biotechnol. 2019 Sep 9. pii: 10.1038/s41587-019-0242-8. doi: 
            10.1038/s41587-019-0242-8.
  PUBMED    31501561
REFERENCE   2  (bases 1 to 15275)
  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 
            Biotechnol. 2019 Sep 9. pii: 10.1038/s41587-019-0242-8. doi: 
            10.1038/s41587-019-0242-8."
COMMENT     Sequence Label: GWF180
FEATURES             Location/Qualifiers
     source          1..15275
                     /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             2054..2083
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2111..2260
                     /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             2267..2287
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2717..3088
                     /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             3107..3676
                     /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"
     misc_feature    join(3694..4267,4268..4270,4271..4279)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3861..3878)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4088..4107
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4301..4900
                     /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(4301..4320)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4797..4817
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4938..5526
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4991..5011)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5409..5420)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5565..5584)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5619..5638
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        5680..6179
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5715..5738)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6112..6131
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6827..7525
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     regulatory      7541..7550
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(7547..7573,7574..7576,7577..10063,10064..10066,
                     10067..11650)
                     /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             7547..11650
                     /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             11654..11680
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             11660..11686
                     /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             11699..11719
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11726..11746
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     repeat_region   11963..12025
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(12076..12096)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(12076..12094)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(12115..12131)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12115..12131)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12128..12150)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12139..12155
                     /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(12163..12169,12170..12187,12188..12193))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12208..12229
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12346..12363)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12517..13105)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12597..12616)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13276..14067,14068..14136))
                     /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     13899..13918
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14137..14241)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(14267..14722)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(14404..14425)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     14616..14635
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     14849..14871
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     14863..14880
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     14864..14880
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     14890..14909
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        14890..14908
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   14938..14972
                     /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 accatgtctt ccgacgaaga
     1381 agaagcgcgc gagctgatcg agcgtgcgaa ggaggctgcg gaacgcgcgc aagaagctgc
     1441 tgagcgtacc ggtgatccgc gcgtgcgtga actcgctcgt gagctgaaac gcctggctca
     1501 ggaagctgct gaagaagtga agcgtgaccc gtcttcctcc gatgtgaacg aagcgctcaa
     1561 actcatcgtg gaagctatcg aggcggctgt ggacgcgctc gaagcggctg aacgcaccgg
     1621 cgacccggag gttcgtgagc tggcgcgtga gctcgtgcgc ctcgcggttg aggctgcgga
     1681 ggaggtgcaa cgtaacccat cctcctctga cgtgaatgag gctctccaca gcatagttta
     1741 cgctattgaa gctgccattt ttgcattgga agctgccgaa aggactggtg acccggaagt
     1801 gagagaactc gcccgagagc ttgtaagact ggcggttgag gcggctgaag aagtccagcg
     1861 aaatccgtcc agccggaatg tcgagcacgc actgatgagg attgtgttgg ccatttacct
     1921 ggcagaagaa aatctccgcg aagctgagga gtctggcgat ccggaaaaac gtgaaaaggc
     1981 tcgcgaacgt gttcgtgagg cggtggaacg tgcggaagag gttcagcgtg atccatccgg
     2041 ttggctgaat catgaacaaa aacttatttc cgaagaggat ctggaattct caagcgctgc
     2101 aggtactagt gatgctttag acgattttga cttagatatg cttggttcag acgcgttaga
     2161 cgacttcgac ctagacatgt taggctcaga tgcattggac gacttcgatt tagatatgtt
     2221 gggctccgat gccctagatg actttgatct agatatgcta ggtagtccca aaaagaagag
     2281 gaaagtggga tcccagtatc tgcccgacac agatgataga caccgaatcg aagagaaacg
     2341 caagcgaacg tatgaaacct tcaaatcgat catgaagaaa tcgcccttct cgggtccgac
     2401 cgatcccagg cccccaccga gaaggattgc ggtcccgtcc cgctcgtcgg ccagcgtgcc
     2461 gaagcctgcg ccgcagccct accccttcac gtcgagcctg agcacaatca attatgacga
     2521 gttcccgacg atggtgttcc cctcgggaca aatctcacaa gcctcggcgc tcgcaccagc
     2581 gcctccccaa gtccttccgc aagcgcctgc cccagcgcct gcaccggcaa tggtgtccgc
     2641 cctcgcacag gcccctgcgc ccgtccccgt gctcgcgcct ggaccgcccc aggcggtcgc
     2701 tccaccggct ccgaagccga cgcaggccgg agagggaaca ctctccgaag cacttcttca
     2761 actccagttt gatgacgagg atcttggagc actccttgga aactcgacag accctgcggt
     2821 gtttaccgac ctcgcgtcag tagataactc cgaatttcag cagcttttga accagggtat
     2881 cccggtcgcg ccacatacaa cggagcccat gttgatggaa taccccgaag caatcacgag
     2941 acttgtgacg ggagcgcagc ggcctcccga tcccgcaccc gcacctttgg gggcacctgg
     3001 cctccctaac ggacttttga gcggcgacga ggatttctcc tccatcgccg atatggattt
     3061 ctcagccttg ctgtcacaga tttccagcgg ctctggcagc ggcagccggg attccaggga
     3121 agggatgttt ttgccgaagc ctgaggccgg ctccgctatt agtgacgtgt ttgagggccg
     3181 cgaggtgtgc cagccaaaac gaatccggcc atttcatcct ccaggaagtc catgggccaa
     3241 ccgcccactc cccgccagcc tcgcaccaac accaaccggt ccagtacatg agccagtcgg
     3301 gtcactgacc ccggcaccag tccctcagcc actggatcca gcgcccgcag tgactcccga
     3361 ggccagtcac ctgttggagg atcccgatga agagacgagc caggctgtca aagcccttcg
     3421 ggagatggcc gatactgtga ttccccagaa ggaagaggct gcaatctgtg gccaaatgga
     3481 cctttcccat ccgcccccaa ggggccatct ggatgagctg acaaccacac ttgagtccat
     3541 gaccgaggat ctgaacctgg actcacccct gaccccggaa ttgaacgaga ttctggatac
     3601 cttcctgaac gacgagtgcc tcttgcatgc catgcatatc agcacaggac tgtccatctt
     3661 cgacacatct ctgttttgaa tttaaataat tcgcccctct ccctcccccc cccctaacgt
     3721 tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac
     3781 catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag
     3841 cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa
     3901 ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag
     3961 gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga
     4021 tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag
     4081 agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc
     4141 ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacgtgtg tttagtcgag
     4201 gttaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga
     4261 tgataatatg gccacaaccg gatccggagc taccatgacc atgaccgagt acaagcccac
     4321 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt
     4381 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac
     4441 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     4501 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     4561 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     4621 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     4681 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     4741 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     4801 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     4861 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga gtttaaacca tgcaagcttg
     4921 atatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     4981 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     5041 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     5101 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     5161 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     5221 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     5281 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     5341 cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     5401 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     5461 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     5521 ccccgcatcc ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5581 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5641 tgtatcttat catgtctgga tcctgcaggt aattctaccg ggtaggggag gcgcttttcc
     5701 caaggcagtc tggagcatgc gctttagcag ccccgctggg cacttggcgc tacacaagtg
     5761 gcctctggcc tcgcacacat tccacatcca ccggtaggcg ccaaccggct ccgttctttg
     5821 gtggcccctt cgcgccacct tctactcctc ccctagtcag gaagttcccc cccgccccgc
     5881 agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg tctcactagg ctcgtgcaga
     5941 tggacagcac cgctgagcaa tggaagcggg taggcctttg gggcagcggc caatagcagc
     6001 tttgctcctt cgctttctgg gctcagaggc tgggaagggg tgggtccggg ggcgggctca
     6061 ggggcgggct caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca
     6121 cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggcctttcga
     6181 cctgcagccc aagcttgtcg acgccaccat gaaaaagaaa ggttctgttg tcatcgttgg
     6241 acgcataaat ctctctggcg acaccgcata ttctcagcag acgcgaggtc tggaagggtg
     6301 tcaagagacc tcacaaaccg ggagggacaa gaatcaagta gaaggggagg tccaggtggt
     6361 atctacggca acccaatcct tcttggcgac atctatcaat ggagtcctgt ggacggttta
     6421 tcatggtgcg ggcactcgga caattgcctc tccaaaaggt ccagttaccc agatgtatac
     6481 taatgtagac aaagatcttg tcggatggca agcaccacaa gggtctcgga gcctcactcc
     6541 ctgtacgtgt ggaagttcag acctttatct cgtaactaga cacgcggatg ttatccccgt
     6601 taggagaaga ggagatagta ggggatccct gctttcacca cgcccaatat cctatcttaa
     6661 gggttcttca ggtggtccct tgttgtgtcc ggcgggtcac gcggtaggta tattccgcgc
     6721 ggccgtatct actcggggcg tggctaaggc ggtcgatttc ataccggttg agtccttgga
     6781 aacaacgatg cggagccccc atatgtcaag cgctgcaggt gctaccatga gcgagctgat
     6841 taaggagaac atgcacatga agctgtacat ggagggcacc gtggacaacc atcacttcaa
     6901 gtgcacatcc gagggcgaag gcaagcccta cgagggcacc cagaccatga gaatcaaggt
     6961 ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg gctactagct tcctctacgg
     7021 cagcaagacc ttcatcaacc acacccaggg catccccgac ttcttcaagc agtccttccc
     7081 tgagggcttc acatgggaga gagtcaccac atacgaagac gggggcgtgc tgaccgctac
     7141 ccaggacacc agcctccagg acggctgcct catctacaac gtcaagatca gaggggtgaa
     7201 cttcacatcc aacggccctg tgatgcagaa gaaaacactc ggctgggagg ccttcaccga
     7261 gacgctgtac cccgctgacg gcggcctgga aggcagaaac gacatggccc tgaagctcgt
     7321 gggcgggagc catctgatcg caaacatcaa gaccacatat agatccaaga aacccgctaa
     7381 gaacctcaag atgcctggcg tctactatgt ggactacaga ctggaaagaa tcaaggaggc
     7441 caacaacgag acctacgtcg agcagcacga ggtggcagtg gccagatact gcgacctccc
     7501 tagcaaactg gggcacaagc ttaattcaag cgctgcaggt gccaccatgg acaagaagta
     7561 cagcatcggc ctggccatcg gcaccaactc tgtgggctgg gccgtgatca ccgacgagta
     7621 caaggtgccc agcaagaaat tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa
     7681 gaacctgatc ggcgccctgc tgttcgacag cggagaaaca gccgaggcca cccggctgaa
     7741 gagaaccgcc agaagaagat acaccagacg gaagaaccgg atctgctatc tgcaagagat
     7801 cttcagcaac gagatggcca aggtggacga cagcttcttc cacagactgg aagagtcctt
     7861 cctggtggaa gaggataaga agcacgagcg gcaccccatc ttcggcaaca tcgtggacga
     7921 ggtggcctac cacgagaagt accccaccat ctaccacctg agaaagaaac tggtggacag
     7981 caccgacaag gccgacctgc ggctgatcta tctggccctg gcccacatga tcaagttccg
     8041 gggccacttc ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt
     8101 catccagctg gtgcagacct acaaccagct gttcgaggaa aaccccatca acgccagcgg
     8161 cgtggacgcc aaggccatcc tgtctgccag actgagcaag agcagacggc tggaaaatct
     8221 gatcgcccag ctgcccggcg agaagaagaa tggcctgttc ggcaacctga ttgccctgag
     8281 cctgggcctg acccccaact tcaagagcaa cttcgacctg gccgaggatg ccaaactgca
     8341 gctgagcaag gacacctacg acgacgacct ggacaacctg ctggcccaga tcggcgacca
     8401 gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc tgagcgacat
     8461 cctgagagtg aacaccgaga tcaccaaggc ccccctgagc gcctctatga tcaagagata
     8521 cgacgagcac caccaggacc tgaccctgct gaaagctctc gtgcggcagc agctgcctga
     8581 gaagtacaaa gagattttct tcgaccagag caagaacggc tacgccggct acatcgatgg
     8641 cggagccagc caggaagagt tctacaagtt catcaagccc atcctggaaa agatggacgg
     8701 caccgaggaa ctgctcgtga agctgaacag agaggacctg ctgcggaagc agcggacctt
     8761 cgacaacggc agcatccccc accagatcca cctgggagag ctgcacgcca ttctgcggcg
     8821 gcaggaagat ttttacccat tcctgaagga caaccgggaa aagatcgaga agatcctgac
     8881 cttccgcatc ccctactacg tgggccctct ggccagggga aacagcagat tcgcctggat
     8941 gaccagaaag agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg tggacaaggg
     9001 cgccagcgcc cagagcttca tcgagcggat gaccaacttc gataagaacc tgcccaacga
     9061 gaaggtgctg cccaagcaca gcctgctgta cgagtacttc accgtgtaca acgagctgac
     9121 caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa
     9181 aaaagccatc gtggacctgc tgttcaagac caaccggaaa gtgaccgtga agcagctgaa
     9241 agaggactac ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga
     9301 tcggttcaac gcctccctgg gcacatacca cgatctgctg aaaattatca aggacaagga
     9361 cttcctggac aatgaggaaa acgaggacat tctggaagat atcgtgctga ccctgacact
     9421 gtttgaggac agagagatga tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga
     9481 caaagtgatg aagcagctga agcggcggag atacaccggc tggggcaggc tgagccggaa
     9541 gctgatcaac ggcatccggg acaagcagtc cggcaagaca atcctggatt tcctgaagtc
     9601 cgacggcttc gccaacagaa acttcatgca gctgatccac gacgacagcc tgacctttaa
     9661 agaggacatc cagaaagccc aggtgtccgg ccagggcgat agcctgcacg agcacattgc
     9721 caatctggcc ggcagccccg ccattaagaa gggcatcctg cagacagtga aggtggtgga
     9781 cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg aaatggccag
     9841 agagaaccag accacccaga agggacagaa gaacagccgc gagagaatga agcggatcga
     9901 agagggcatc aaagagctgg gcagccagat cctgaaagaa caccccgtgg aaaacaccca
     9961 gctgcagaac gagaagctgt acctgtacta cctgcagaat gggcgggata tgtacgtgga
    10021 ccaggaactg gacatcaacc ggctgtccga ctacgatgtg gacgctatcg tgcctcagag
    10081 ctttctgaag gacgactcca tcgataacaa agtgctgact cggagcgaca agaaccgggg
    10141 caagagcgac aacgtgccct ccgaagaggt cgtgaagaag atgaagaact actggcgcca
    10201 gctgctgaat gccaagctga ttacccagag gaagttcgac aatctgacca aggccgagag
    10261 aggcggcctg agcgaactgg ataaggccgg cttcatcaag agacagctgg tggaaacccg
    10321 gcagatcaca aagcacgtgg cacagatcct ggactcccgg atgaacacta agtacgacga
    10381 gaacgacaaa ctgatccggg aagtgaaagt gatcaccctg aagtccaagc tggtgtccga
    10441 tttccggaag gatttccagt tttacaaagt gcgcgagatc aacaactacc accacgccca
    10501 cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga
    10561 aagcgagttc gtgtacggcg actacaaggt gtacgacgtg cggaagatga tcgccaagag
    10621 cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc tacagcaaca tcatgaactt
    10681 tttcaagacc gagattaccc tggccaacgg cgagatccgg aagcggcctc tgatcgagac
    10741 aaacggcgaa acaggcgaga tcgtgtggga taagggccgg gactttgcca ccgtgcggaa
    10801 agtgctgtct atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt
    10861 cagcaaagag tctatcctgc ccaagaggaa cagcgacaag ctgatcgcca gaaagaagga
    10921 ctgggaccct aagaagtacg gcggcttcga cagccccacc gtggcctatt ctgtgctggt
    10981 ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag agctgctggg
    11041 gatcaccatc atggaaagaa gcagcttcga gaagaatccc atcgactttc tggaagccaa
    11101 gggctacaaa gaagtgaaaa aggacctgat catcaagctg cctaagtact ccctgttcga
    11161 gctggaaaac ggccggaaga gaatgctggc ctctgccggc gaactgcaga agggaaacga
    11221 actggccctg ccctccaaat atgtgaactt cctgtacctg gccagccact atgagaagct
    11281 gaagggctcc cccgaggata atgagcagaa acagctgttt gtggaacagc acaaacacta
    11341 cctggacgag atcatcgagc agatcagcga gttctccaag agagtgatcc tggccgacgc
    11401 taatctggac aaggtgctga gcgcctacaa caagcacaga gacaagccta tcagagagca
    11461 ggccgagaat atcatccacc tgtttaccct gaccaatctg ggagcccctg ccgccttcaa
    11521 gtactttgac accaccatcg accggaagag gtacaccagc accaaagagg tgctggacgc
    11581 caccctgatc caccagagca tcaccggcct gtacgagaca cggatcgacc tgtctcagct
    11641 gggaggcgac gcctatccct atgacgtgcc cgattatgcc agcctgggca gcggctcccc
    11701 caagaaaaaa cgcaaggtgg aagatcctaa gaaaaagcgg aaagtggacg gcattggtag
    11761 tgggagcaac ggatgactcg agctagttaa aagttttgtt actttataga agaaattttg
    11821 agtttttgtt tttttttaat aaataaataa acataaataa attgtttgtt gaatttatta
    11881 ttagtatgta agtgtaaata taataaaact taatatctat tcaaattaat aaataaacct
    11941 cgatatacag accgataaaa cacatgcgtc aattttacgc atgattatct ttaacgtacg
    12001 tcacaatatg attatctttc tagggttaat ctagtatacg cgtatgcggc cgcttaatta
    12061 atccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata
    12121 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
    12181 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
    12241 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
    12301 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
    12361 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
    12421 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
    12481 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
    12541 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
    12601 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
    12661 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
    12721 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
    12781 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
    12841 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
    12901 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
    12961 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
    13021 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
    13081 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
    13141 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
    13201 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
    13261 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
    13321 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
    13381 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
    13441 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
    13501 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
    13561 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
    13621 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
    13681 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
    13741 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
    13801 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
    13861 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
    13921 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
    13981 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
    14041 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
    14101 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
    14161 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
    14221 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt
    14281 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
    14341 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
    14401 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
    14461 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
    14521 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
    14581 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
    14641 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
    14701 aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg
    14761 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
    14821 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
    14881 gagcgcgcgt aatacgactc actatagggc gaattggggc gcgccattct agattaaccc
    14941 tagaaagata gtctgcgtaa aattgacgca tgcattcttg aaatattgct ctctctttct
    15001 aaatagcgcg aatccgtcgc tgtgcattta ggacatctca gtcgccgctt ggagctcccg
    15061 tgaggcgtgc ttgtcaatgc ggtaagtgtc actgattttg aactataacg accgcgtgag
    15121 tcaaaatgac gcatgattat cttttacgtg acttttaaga tttaactcat acgataatta
    15181 tattgttatt tcatgttcta cttacgtgat aacttattat atatatattt tcttgttata
    15241 gatatcaact agaatgctag catgggccca gatct
//
