LOCUS       sequence-261658-       15273 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 15273)
  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 15273)
  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..15273
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..284
                     /label=chicken beta-actin promoter
     intron          285..1301
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1225..1247
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1309..1328
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2052..2081
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2109..2258
                     /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             2265..2285
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2715..3086
                     /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             3105..3674
                     /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(3692..4265,4266..4268,4269..4277)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3859..3876)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4086..4105
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4299..4898
                     /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(4299..4318)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4795..4815
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4936..5524
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4989..5009)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5407..5418)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5563..5582)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5617..5636
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        5678..6177
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5713..5736)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6110..6129
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6825..7523
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     regulatory      7539..7548
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(7545..7571,7572..7574,7575..10061,10062..10064,
                     10065..11648)
                     /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             7545..11648
                     /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             11652..11678
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             11658..11684
                     /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             11697..11717
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11724..11744
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     repeat_region   11961..12023
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(12074..12094)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(12074..12092)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(12113..12129)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12113..12129)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12126..12148)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12137..12153
                     /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(12161..12167,12168..12185,12186..12191))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12206..12227
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12344..12361)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12515..13103)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12595..12614)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13274..14065,14066..14134))
                     /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     13897..13916
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14135..14239)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(14265..14720)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(14402..14423)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     14614..14633
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     14847..14869
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     14861..14878
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     14862..14878
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     14888..14907
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        14888..14906
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   14936..14970
                     /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 ggcgggagtc gctgcgcgct
      301 gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga
      361 ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc
      421 gcttggttta atgacggctt gtttcttttc tgtggctgcg tgaaagcctt gaggggctcc
      481 gggagggccc tttgtgcggg gggagcggct cggggggtgc gtgcgtgtgt gtgtgcgtgg
      541 ggagcgccgc gtgcggctcc gcgctgcccg gcggctgtga gcgctgcggg cgcggcgcgg
      601 ggctttgtgc gctccgcagt gtgcgcgagg ggagcgcggc cgggggcggt gccccgcggt
      661 gcgggggggg ctgcgagggg aacaaaggct gcgtgcgggg tgtgtgcgtg ggggggtgag
      721 cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct gcacccccct ccccgagttg
      781 ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg gcgtggcgcg gggctcgccg
      841 tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg cctcgggccg
      901 gggagggctc gggggagggg cgcggcggcc cccggagcgc cggcggctgt cgaggcgcgg
      961 cgagccgcag ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga cttcctttgt
     1021 cccaaatctg tgcggagccg aaatctggga ggcgccgccg caccccctct agcgggcgcg
     1081 gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt cgtgcgtcgc
     1141 cgcgccgccg tccccttctc cctctccagc ctcggggctg tccgcggggg gacggctgcc
     1201 ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc ggctctagag
     1261 cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc aacgtgctgg
     1321 ttattgtgct gtctcatcat tttggcaaag aattcgccac catgtcttcc gacgaagaag
     1381 aagcgcgcga gctgatcgag cgtgcgaagg aggctgcgga acgcgcgcaa gaagctgctg
     1441 agcgtaccgg tgatccgcgc gtgcgtgaac tcgctcgtga gctgaaacgc ctggctcagg
     1501 aagctgctga agaagtgaag cgtgacccgt cttcctccga tgtgaacgaa gcgctcaaac
     1561 tcatcgtgga agctatcgag gcggctgtgg acgcgctcga agcggctgaa cgcaccggcg
     1621 acccggaggt tcgtgagctg gcgcgtgagc tcgtgcgcct cgcggttgag gctgcggagg
     1681 aggtgcaacg taacccatcc tcctctgacg tgaatgaggc tctccacagc atagtttacg
     1741 ctattgaagc tgccattttt gcattggaag ctgccgaaag gactggtgac ccggaagtga
     1801 gagaactcgc ccgagagctt gtaagactgg cggttgaggc ggctgaagaa gtccagcgaa
     1861 atccgtccag ccggaatgtc gagcacgcac tgatgaggat tgtgttggcc atttacctgg
     1921 cagaagaaaa tctccgcgaa gctgaggagt ctggcgatcc ggaaaaacgt gaaaaggctc
     1981 gcgaacgtgt tcgtgaggcg gtggaacgtg cggaagaggt tcagcgtgat ccatccggtt
     2041 ggctgaatca tgaacaaaaa cttatttccg aagaggatct ggaattctca agcgctgcag
     2101 gtactagtga tgctttagac gattttgact tagatatgct tggttcagac gcgttagacg
     2161 acttcgacct agacatgtta ggctcagatg cattggacga cttcgattta gatatgttgg
     2221 gctccgatgc cctagatgac tttgatctag atatgctagg tagtcccaaa aagaagagga
     2281 aagtgggatc ccagtatctg cccgacacag atgatagaca ccgaatcgaa gagaaacgca
     2341 agcgaacgta tgaaaccttc aaatcgatca tgaagaaatc gcccttctcg ggtccgaccg
     2401 atcccaggcc cccaccgaga aggattgcgg tcccgtcccg ctcgtcggcc agcgtgccga
     2461 agcctgcgcc gcagccctac cccttcacgt cgagcctgag cacaatcaat tatgacgagt
     2521 tcccgacgat ggtgttcccc tcgggacaaa tctcacaagc ctcggcgctc gcaccagcgc
     2581 ctccccaagt ccttccgcaa gcgcctgccc cagcgcctgc accggcaatg gtgtccgccc
     2641 tcgcacaggc ccctgcgccc gtccccgtgc tcgcgcctgg accgccccag gcggtcgctc
     2701 caccggctcc gaagccgacg caggccggag agggaacact ctccgaagca cttcttcaac
     2761 tccagtttga tgacgaggat cttggagcac tccttggaaa ctcgacagac cctgcggtgt
     2821 ttaccgacct cgcgtcagta gataactccg aatttcagca gcttttgaac cagggtatcc
     2881 cggtcgcgcc acatacaacg gagcccatgt tgatggaata ccccgaagca atcacgagac
     2941 ttgtgacggg agcgcagcgg cctcccgatc ccgcacccgc acctttgggg gcacctggcc
     3001 tccctaacgg acttttgagc ggcgacgagg atttctcctc catcgccgat atggatttct
     3061 cagccttgct gtcacagatt tccagcggct ctggcagcgg cagccgggat tccagggaag
     3121 ggatgttttt gccgaagcct gaggccggct ccgctattag tgacgtgttt gagggccgcg
     3181 aggtgtgcca gccaaaacga atccggccat ttcatcctcc aggaagtcca tgggccaacc
     3241 gcccactccc cgccagcctc gcaccaacac caaccggtcc agtacatgag ccagtcgggt
     3301 cactgacccc ggcaccagtc cctcagccac tggatccagc gcccgcagtg actcccgagg
     3361 ccagtcacct gttggaggat cccgatgaag agacgagcca ggctgtcaaa gcccttcggg
     3421 agatggccga tactgtgatt ccccagaagg aagaggctgc aatctgtggc caaatggacc
     3481 tttcccatcc gcccccaagg ggccatctgg atgagctgac aaccacactt gagtccatga
     3541 ccgaggatct gaacctggac tcacccctga ccccggaatt gaacgagatt ctggatacct
     3601 tcctgaacga cgagtgcctc ttgcatgcca tgcatatcag cacaggactg tccatcttcg
     3661 acacatctct gttttgaatt taaataattc gcccctctcc ctcccccccc cctaacgtta
     3721 ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca
     3781 tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca
     3841 ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg
     3901 aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc
     3961 agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata
     4021 cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag
     4081 tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc
     4141 attgtatggg atctgatctg gggcctcggt gcacatgctt tacgtgtgtt tagtcgaggt
     4201 taaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg
     4261 ataatatggc cacaaccgga tccggagcta ccatgaccat gaccgagtac aagcccacgg
     4321 tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg
     4381 ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg
     4441 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg
     4501 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt
     4561 tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac
     4621 agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg
     4681 tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag
     4741 tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc
     4801 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac
     4861 cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgagt ttaaaccatg caagcttgat
     4921 atcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     4981 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     5041 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     5101 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     5161 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     5221 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     5281 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     5341 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     5401 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     5461 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     5521 ccgcatcctt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5581 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5641 tatcttatca tgtctggatc ctgcaggtaa ttctaccggg taggggaggc gcttttccca
     5701 aggcagtctg gagcatgcgc tttagcagcc ccgctgggca cttggcgcta cacaagtggc
     5761 ctctggcctc gcacacattc cacatccacc ggtaggcgcc aaccggctcc gttctttggt
     5821 ggccccttcg cgccaccttc tactcctccc ctagtcagga agttcccccc cgccccgcag
     5881 ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc tcactaggct cgtgcagatg
     5941 gacagcaccg ctgagcaatg gaagcgggta ggcctttggg gcagcggcca atagcagctt
     6001 tgctccttcg ctttctgggc tcagaggctg ggaaggggtg ggtccggggg cgggctcagg
     6061 ggcgggctca ggggcggggc gggcgcccga aggtcctccg gaggcccggc attctgcacg
     6121 cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct catctccggg cctttcgacc
     6181 tgcagcccaa gcttgtcgac gccaccatga aaaagaaagg ttctgttgtc atcgttggac
     6241 gcataaatct ctctggcgac accgcatatt ctcagcagac gcgaggtctg gaagggtgtc
     6301 aagagacctc acaaaccggg agggacaaga atcaagtaga aggggaggtc caggtggtat
     6361 ctacggcaac ccaatccttc ttggcgacat ctatcaatgg agtcctgtgg acggtttatc
     6421 atggtgcggg cactcggaca attgcctctc caaaaggtcc agttacccag atgtatacta
     6481 atgtagacaa agatcttgtc ggatggcaag caccacaagg gtctcggagc ctcactccct
     6541 gtacgtgtgg aagttcagac ctttatctcg taactagaca cgcggatgtt atccccgtta
     6601 ggagaagagg agatagtagg ggatccctgc tttcaccacg cccaatatcc tatcttaagg
     6661 gttcttcagg tggtcccttg ttgtgtccgg cgggtcacgc ggtaggtata ttccgcgcgg
     6721 ccgtatctac tcggggcgtg gctaaggcgg tcgatttcat accggttgag tccttggaaa
     6781 caacgatgcg gagcccccat atgtcaagcg ctgcaggtgc taccatgagc gagctgatta
     6841 aggagaacat gcacatgaag ctgtacatgg agggcaccgt ggacaaccat cacttcaagt
     6901 gcacatccga gggcgaaggc aagccctacg agggcaccca gaccatgaga atcaaggtgg
     6961 tcgagggcgg ccctctcccc ttcgccttcg acatcctggc tactagcttc ctctacggca
     7021 gcaagacctt catcaaccac acccagggca tccccgactt cttcaagcag tccttccctg
     7081 agggcttcac atgggagaga gtcaccacat acgaagacgg gggcgtgctg accgctaccc
     7141 aggacaccag cctccaggac ggctgcctca tctacaacgt caagatcaga ggggtgaact
     7201 tcacatccaa cggccctgtg atgcagaaga aaacactcgg ctgggaggcc ttcaccgaga
     7261 cgctgtaccc cgctgacggc ggcctggaag gcagaaacga catggccctg aagctcgtgg
     7321 gcgggagcca tctgatcgca aacatcaaga ccacatatag atccaagaaa cccgctaaga
     7381 acctcaagat gcctggcgtc tactatgtgg actacagact ggaaagaatc aaggaggcca
     7441 acaacgagac ctacgtcgag cagcacgagg tggcagtggc cagatactgc gacctcccta
     7501 gcaaactggg gcacaagctt aattcaagcg ctgcaggtgc caccatggac aagaagtaca
     7561 gcatcggcct ggccatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca
     7621 aggtgcccag caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga
     7681 acctgatcgg cgccctgctg ttcgacagcg gagaaacagc cgaggccacc cggctgaaga
     7741 gaaccgccag aagaagatac accagacgga agaaccggat ctgctatctg caagagatct
     7801 tcagcaacga gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc
     7861 tggtggaaga ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg
     7921 tggcctacca cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca
     7981 ccgacaaggc cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg
     8041 gccacttcct gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca
     8101 tccagctggt gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg
     8161 tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga
     8221 tcgcccagct gcccggcgag aagaagaatg gcctgttcgg caacctgatt gccctgagcc
     8281 tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc
     8341 tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt
     8401 acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc
     8461 tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg
     8521 acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga
     8581 agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac atcgatggcg
     8641 gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca
     8701 ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg
     8761 acaacggcag catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc
     8821 aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct
     8881 tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga
     8941 ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg
     9001 ccagcgccca gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga
     9061 aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtacaac gagctgacca
     9121 aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa
     9181 aagccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag
     9241 aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc
     9301 ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact
     9361 tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt
     9421 ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca
     9481 aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc
     9541 tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg
     9601 acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag
     9661 aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca
     9721 atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg
     9781 agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag
     9841 agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag
     9901 agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc
     9961 tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc
    10021 aggaactgga catcaaccgg ctgtccgact acgatgtgga cgctatcgtg cctcagagct
    10081 ttctgaagga cgactccatc gataacaaag tgctgactcg gagcgacaag aaccggggca
    10141 agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcgccagc
    10201 tgctgaatgc caagctgatt acccagagga agttcgacaa tctgaccaag gccgagagag
    10261 gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc
    10321 agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga
    10381 acgacaaact gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt
    10441 tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg
    10501 acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa
    10561 gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg
    10621 agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt
    10681 tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa
    10741 acggcgaaac aggcgagatc gtgtgggata agggccggga ctttgccacc gtgcggaaag
    10801 tgctgtctat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca
    10861 gcaaagagtc tatcctgccc aagaggaaca gcgacaagct gatcgccaga aagaaggact
    10921 gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct gtgctggtgg
    10981 tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctgctgggga
    11041 tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg
    11101 gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc
    11161 tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag ggaaacgaac
    11221 tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga
    11281 agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac aaacactacc
    11341 tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta
    11401 atctggacaa ggtgctgagc gcctacaaca agcacagaga caagcctatc agagagcagg
    11461 ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt
    11521 actttgacac caccatcgac cggaagaggt acaccagcac caaagaggtg ctggacgcca
    11581 ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg
    11641 gaggcgacgc ctatccctat gacgtgcccg attatgccag cctgggcagc ggctccccca
    11701 agaaaaaacg caaggtggaa gatcctaaga aaaagcggaa agtggacggc attggtagtg
    11761 ggagcaacgg atgactcgag ctagttaaaa gttttgttac tttatagaag aaattttgag
    11821 tttttgtttt tttttaataa ataaataaac ataaataaat tgtttgttga atttattatt
    11881 agtatgtaag tgtaaatata ataaaactta atatctattc aaattaataa ataaacctcg
    11941 atatacagac cgataaaaca catgcgtcaa ttttacgcat gattatcttt aacgtacgtc
    12001 acaatatgat tatctttcta gggttaatct agtatacgcg tatgcggccg cttaattaat
    12061 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc
    12121 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
    12181 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
    12241 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
    12301 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
    12361 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
    12421 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
    12481 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
    12541 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
    12601 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
    12661 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
    12721 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
    12781 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
    12841 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
    12901 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
    12961 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
    13021 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
    13081 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
    13141 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
    13201 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
    13261 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
    13321 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
    13381 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
    13441 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
    13501 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
    13561 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
    13621 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
    13681 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
    13741 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
    13801 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
    13861 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
    13921 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
    13981 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
    14041 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
    14101 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
    14161 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
    14221 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa
    14281 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
    14341 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
    14401 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
    14461 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
    14521 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
    14581 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
    14641 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
    14701 tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg
    14761 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
    14821 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
    14881 gcgcgcgtaa tacgactcac tatagggcga attggggcgc gccattctag attaacccta
    14941 gaaagatagt ctgcgtaaaa ttgacgcatg cattcttgaa atattgctct ctctttctaa
    15001 atagcgcgaa tccgtcgctg tgcatttagg acatctcagt cgccgcttgg agctcccgtg
    15061 aggcgtgctt gtcaatgcgg taagtgtcac tgattttgaa ctataacgac cgcgtgagtc
    15121 aaaatgacgc atgattatct tttacgtgac ttttaagatt taactcatac gataattata
    15181 ttgttatttc atgttctact tacgtgataa cttattatat atatattttc ttgttataga
    15241 tatcaactag aatgctagca tgggcccaga tct
//
