LOCUS       sequence-160812-       15142 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GA-inducible VPR-Sp dCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15142)
  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 15142)
  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: pSLQ2812 pPB: CAG-GID1-VPR-IRES-Puro-WPRE
            PGK-...
FEATURES             Location/Qualifiers
     source          1..15142
                     /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"
     regulatory      1358..1367
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2426..2575
                     /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             2582..2602
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3032..3403
                     /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             3422..3991
                     /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(4009..4582,4583..4585,4586..4594)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4176..4193)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4403..4422
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4616..5215
                     /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(4616..4635)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5112..5132
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    5253..5841
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5306..5326)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5724..5735)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        5862..6361
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5897..5920)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6294..6313
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6403..6420
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             6694..7392
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     regulatory      7408..7417
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(7414..7440,7441..7443,7444..9930,9931..9933,
                     9934..11517)
                     /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             7414..11517
                     /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             11521..11547
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             11527..11553
                     /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             11566..11586
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11593..11613
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     repeat_region   11830..11892
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(11943..11963)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(11943..11961)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(11982..11998)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11982..11998)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11995..12017)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12006..12022
                     /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(12030..12036,12037..12054,12055..12060))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12075..12096
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12213..12230)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12384..12972)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12464..12483)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13143..13934,13935..14003))
                     /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     13766..13785
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14004..14108)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(14134..14589)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(14271..14292)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     14483..14502
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     14716..14738
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     14730..14747
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     14731..14747
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     14757..14776
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        14757..14775
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   14805..14839
                     /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 accatggctg cgagcgatga
     1381 agttaatctt attgagagca gaacagtggt tcctctcaat acatgggttt taatatccaa
     1441 cttcaaagta gcctacaata tccttcgtcg ccctgatgga acctttaacc gacacttagc
     1501 tgagtatcta gaccgtaaag tcactgcaaa cgccaatccg gttgatgggg ttttctcgtt
     1561 cgatgtcttg attgatcgca ggatcaatct tctaagcaga gtctatagac cagcttatgc
     1621 agatcaagag caacctccta gtattttaga tctcgagaag cctgttgatg gcgacattgt
     1681 ccctgttata ttgttcttcc atggaggtag ctttgctcat tcttctgcaa acagtgccat
     1741 ctacgatact ctttgtcgca ggcttgttgg tttgtgcaag tgtgttgttg tctctgtgaa
     1801 ttatcggcgt gcaccagaga atccataccc ttgtgcttat gatgatggtt ggattgctct
     1861 taattgggtt aactcgagat cttggcttaa atccaagaaa gactcaaagg tccatatttt
     1921 cttggctggt gatagctctg gaggtaacat cgcgcataat gtggctttaa gagcgggtga
     1981 atcgggaatc gatgttttgg ggaacattct gctgaatcct atgtttggtg ggaatgagag
     2041 aacggagtct gagaaaagtt tggatgggaa atactttgtg acggttagag accgcgattg
     2101 gtactggaaa gcgtttttac ccgagggaga agatagagag catccagcgt gtaatccgtt
     2161 tagcccgaga gggaaaagct tagaaggagt gagtttcccc aagagtcttg tggttgtcgc
     2221 gggtttggat ttgattagag attggcagtt ggcatacgcg gaagggctca agaaagcggg
     2281 tcaagaggtt aagcttatgc atttagagaa agcaactgtt gggttttacc tcttgcctaa
     2341 taacaatcat ttccataatg ttatggatga gatttcggcg tttgtaaacg cggaatgtga
     2401 attctcaagc gctgcaggta ctagtgatgc tttagacgat tttgacttag atatgcttgg
     2461 ttcagacgcg ttagacgact tcgacctaga catgttaggc tcagatgcat tggacgactt
     2521 cgatttagat atgttgggct ccgatgccct agatgacttt gatctagata tgctaggtag
     2581 tcccaaaaag aagaggaaag tgggatccca gtatctgccc gacacagatg atagacaccg
     2641 aatcgaagag aaacgcaagc gaacgtatga aaccttcaaa tcgatcatga agaaatcgcc
     2701 cttctcgggt ccgaccgatc ccaggccccc accgagaagg attgcggtcc cgtcccgctc
     2761 gtcggccagc gtgccgaagc ctgcgccgca gccctacccc ttcacgtcga gcctgagcac
     2821 aatcaattat gacgagttcc cgacgatggt gttcccctcg ggacaaatct cacaagcctc
     2881 ggcgctcgca ccagcgcctc cccaagtcct tccgcaagcg cctgccccag cgcctgcacc
     2941 ggcaatggtg tccgccctcg cacaggcccc tgcgcccgtc cccgtgctcg cgcctggacc
     3001 gccccaggcg gtcgctccac cggctccgaa gccgacgcag gccggagagg gaacactctc
     3061 cgaagcactt cttcaactcc agtttgatga cgaggatctt ggagcactcc ttggaaactc
     3121 gacagaccct gcggtgttta ccgacctcgc gtcagtagat aactccgaat ttcagcagct
     3181 tttgaaccag ggtatcccgg tcgcgccaca tacaacggag cccatgttga tggaataccc
     3241 cgaagcaatc acgagacttg tgacgggagc gcagcggcct cccgatcccg cacccgcacc
     3301 tttgggggca cctggcctcc ctaacggact tttgagcggc gacgaggatt tctcctccat
     3361 cgccgatatg gatttctcag ccttgctgtc acagatttcc agcggctctg gcagcggcag
     3421 ccgggattcc agggaaggga tgtttttgcc gaagcctgag gccggctccg ctattagtga
     3481 cgtgtttgag ggccgcgagg tgtgccagcc aaaacgaatc cggccatttc atcctccagg
     3541 aagtccatgg gccaaccgcc cactccccgc cagcctcgca ccaacaccaa ccggtccagt
     3601 acatgagcca gtcgggtcac tgaccccggc accagtccct cagccactgg atccagcgcc
     3661 cgcagtgact cccgaggcca gtcacctgtt ggaggatccc gatgaagaga cgagccaggc
     3721 tgtcaaagcc cttcgggaga tggccgatac tgtgattccc cagaaggaag aggctgcaat
     3781 ctgtggccaa atggaccttt cccatccgcc cccaaggggc catctggatg agctgacaac
     3841 cacacttgag tccatgaccg aggatctgaa cctggactca cccctgaccc cggaattgaa
     3901 cgagattctg gataccttcc tgaacgacga gtgcctcttg catgccatgc atatcagcac
     3961 aggactgtcc atcttcgaca catctctgtt ttgaatttaa ataattcgcc cctctccctc
     4021 ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta
     4081 tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc
     4141 tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct
     4201 gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt
     4261 agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa
     4321 gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg
     4381 gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga
     4441 tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac
     4501 gtgtgtttag tcgaggttaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     4561 ctttgaaaaa cacgatgata atatggccac aaccggatcc ggagctacca tgaccatgac
     4621 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac
     4681 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca
     4741 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg
     4801 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt
     4861 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg
     4921 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc
     4981 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc
     5041 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac
     5101 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt
     5161 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgagttta
     5221 aaccatgcaa gcttgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg
     5281 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     5341 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     5401 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     5461 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     5521 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     5581 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     5641 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg
     5701 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     5761 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     5821 ccctttgggc cgcctccccg catcctgcag gtaattctac cgggtagggg aggcgctttt
     5881 cccaaggcag tctggagcat gcgctttagc agccccgctg ggcacttggc gctacacaag
     5941 tggcctctgg cctcgcacac attccacatc caccggtagg cgccaaccgg ctccgttctt
     6001 tggtggcccc ttcgcgccac cttctactcc tcccctagtc aggaagttcc cccccgcccc
     6061 gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta ggctcgtgca
     6121 gatggacagc accgctgagc aatggaagcg ggtaggcctt tggggcagcg gccaatagca
     6181 gctttgctcc ttcgctttct gggctcagag gctgggaagg ggtgggtccg ggggcgggct
     6241 caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg
     6301 cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct tcctcatctc cgggcctttc
     6361 gacctgcagc ccaagcttgt cgacgccacc atgaagagag atcatcatca tcatcatcat
     6421 caagataaga agactatgat gatgaatgaa gaagacgacg gtaacggcat ggatgagctt
     6481 ctagctgttc ttggttacaa ggttaggtca tccgaaatgg ctgatgttgc tcagaaactc
     6541 gagcagcttg aagttatgat gtctaatgtt caagaagacg atctttctca actcgctact
     6601 gagactgttc actataatcc ggcggagctt tacacgtggc ttgattctat gctcaccgac
     6661 cttaatcata tgtcaagcgc tgcaggtgct accatgagcg agctgattaa ggagaacatg
     6721 cacatgaagc tgtacatgga gggcaccgtg gacaaccatc acttcaagtg cacatccgag
     6781 ggcgaaggca agccctacga gggcacccag accatgagaa tcaaggtggt cgagggcggc
     6841 cctctcccct tcgccttcga catcctggct actagcttcc tctacggcag caagaccttc
     6901 atcaaccaca cccagggcat ccccgacttc ttcaagcagt ccttccctga gggcttcaca
     6961 tgggagagag tcaccacata cgaagacggg ggcgtgctga ccgctaccca ggacaccagc
     7021 ctccaggacg gctgcctcat ctacaacgtc aagatcagag gggtgaactt cacatccaac
     7081 ggccctgtga tgcagaagaa aacactcggc tgggaggcct tcaccgagac gctgtacccc
     7141 gctgacggcg gcctggaagg cagaaacgac atggccctga agctcgtggg cgggagccat
     7201 ctgatcgcaa acatcaagac cacatataga tccaagaaac ccgctaagaa cctcaagatg
     7261 cctggcgtct actatgtgga ctacagactg gaaagaatca aggaggccaa caacgagacc
     7321 tacgtcgagc agcacgaggt ggcagtggcc agatactgcg acctccctag caaactgggg
     7381 cacaagctta attcaagcgc tgcaggtgcc accatggaca agaagtacag catcggcctg
     7441 gccatcggca ccaactctgt gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc
     7501 aagaaattca aggtgctggg caacaccgac cggcacagca tcaagaagaa cctgatcggc
     7561 gccctgctgt tcgacagcgg agaaacagcc gaggccaccc ggctgaagag aaccgccaga
     7621 agaagataca ccagacggaa gaaccggatc tgctatctgc aagagatctt cagcaacgag
     7681 atggccaagg tggacgacag cttcttccac agactggaag agtccttcct ggtggaagag
     7741 gataagaagc acgagcggca ccccatcttc ggcaacatcg tggacgaggt ggcctaccac
     7801 gagaagtacc ccaccatcta ccacctgaga aagaaactgg tggacagcac cgacaaggcc
     7861 gacctgcggc tgatctatct ggccctggcc cacatgatca agttccgggg ccacttcctg
     7921 atcgagggcg acctgaaccc cgacaacagc gacgtggaca agctgttcat ccagctggtg
     7981 cagacctaca accagctgtt cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag
     8041 gccatcctgt ctgccagact gagcaagagc agacggctgg aaaatctgat cgcccagctg
     8101 cccggcgaga agaagaatgg cctgttcggc aacctgattg ccctgagcct gggcctgacc
     8161 cccaacttca agagcaactt cgacctggcc gaggatgcca aactgcagct gagcaaggac
     8221 acctacgacg acgacctgga caacctgctg gcccagatcg gcgaccagta cgccgacctg
     8281 tttctggccg ccaagaacct gtccgacgcc atcctgctga gcgacatcct gagagtgaac
     8341 accgagatca ccaaggcccc cctgagcgcc tctatgatca agagatacga cgagcaccac
     8401 caggacctga ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa gtacaaagag
     8461 attttcttcg accagagcaa gaacggctac gccggctaca tcgatggcgg agccagccag
     8521 gaagagttct acaagttcat caagcccatc ctggaaaaga tggacggcac cgaggaactg
     8581 ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc ggaccttcga caacggcagc
     8641 atcccccacc agatccacct gggagagctg cacgccattc tgcggcggca ggaagatttt
     8701 tacccattcc tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc
     8761 tactacgtgg gccctctggc caggggaaac agcagattcg cctggatgac cagaaagagc
     8821 gaggaaacca tcaccccctg gaacttcgag gaagtggtgg acaagggcgc cagcgcccag
     8881 agcttcatcg agcggatgac caacttcgat aagaacctgc ccaacgagaa ggtgctgccc
     8941 aagcacagcc tgctgtacga gtacttcacc gtgtacaacg agctgaccaa agtgaaatac
     9001 gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa agccatcgtg
     9061 gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga ggactacttc
     9121 aagaaaatcg agtgcttcga ctccgtggaa atctccggcg tggaagatcg gttcaacgcc
     9181 tccctgggca cataccacga tctgctgaaa attatcaagg acaaggactt cctggacaat
     9241 gaggaaaacg aggacattct ggaagatatc gtgctgaccc tgacactgtt tgaggacaga
     9301 gagatgatcg aggaacggct gaaaacctat gcccacctgt tcgacgacaa agtgatgaag
     9361 cagctgaagc ggcggagata caccggctgg ggcaggctga gccggaagct gatcaacggc
     9421 atccgggaca agcagtccgg caagacaatc ctggatttcc tgaagtccga cggcttcgcc
     9481 aacagaaact tcatgcagct gatccacgac gacagcctga cctttaaaga ggacatccag
     9541 aaagcccagg tgtccggcca gggcgatagc ctgcacgagc acattgccaa tctggccggc
     9601 agccccgcca ttaagaaggg catcctgcag acagtgaagg tggtggacga gctcgtgaaa
     9661 gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga gaaccagacc
     9721 acccagaagg gacagaagaa cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa
     9781 gagctgggca gccagatcct gaaagaacac cccgtggaaa acacccagct gcagaacgag
     9841 aagctgtacc tgtactacct gcagaatggg cgggatatgt acgtggacca ggaactggac
     9901 atcaaccggc tgtccgacta cgatgtggac gctatcgtgc ctcagagctt tctgaaggac
     9961 gactccatcg ataacaaagt gctgactcgg agcgacaaga accggggcaa gagcgacaac
    10021 gtgccctccg aagaggtcgt gaagaagatg aagaactact ggcgccagct gctgaatgcc
    10081 aagctgatta cccagaggaa gttcgacaat ctgaccaagg ccgagagagg cggcctgagc
    10141 gaactggata aggccggctt catcaagaga cagctggtgg aaacccggca gatcacaaag
    10201 cacgtggcac agatcctgga ctcccggatg aacactaagt acgacgagaa cgacaaactg
    10261 atccgggaag tgaaagtgat caccctgaag tccaagctgg tgtccgattt ccggaaggat
    10321 ttccagtttt acaaagtgcg cgagatcaac aactaccacc acgcccacga cgcctacctg
    10381 aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctggaaag cgagttcgtg
    10441 tacggcgact acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc
    10501 ggcaaggcta ccgccaagta cttcttctac agcaacatca tgaacttttt caagaccgag
    10561 attaccctgg ccaacggcga gatccggaag cggcctctga tcgagacaaa cggcgaaaca
    10621 ggcgagatcg tgtgggataa gggccgggac tttgccaccg tgcggaaagt gctgtctatg
    10681 ccccaagtga atatcgtgaa aaagaccgag gtgcagacag gcggcttcag caaagagtct
    10741 atcctgccca agaggaacag cgacaagctg atcgccagaa agaaggactg ggaccctaag
    10801 aagtacggcg gcttcgacag ccccaccgtg gcctattctg tgctggtggt ggccaaagtg
    10861 gaaaagggca agtccaagaa actgaagagt gtgaaagagc tgctggggat caccatcatg
    10921 gaaagaagca gcttcgagaa gaatcccatc gactttctgg aagccaaggg ctacaaagaa
    10981 gtgaaaaagg acctgatcat caagctgcct aagtactccc tgttcgagct ggaaaacggc
    11041 cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact ggccctgccc
    11101 tccaaatatg tgaacttcct gtacctggcc agccactatg agaagctgaa gggctccccc
    11161 gaggataatg agcagaaaca gctgtttgtg gaacagcaca aacactacct ggacgagatc
    11221 atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa tctggacaag
    11281 gtgctgagcg cctacaacaa gcacagagac aagcctatca gagagcaggc cgagaatatc
    11341 atccacctgt ttaccctgac caatctggga gcccctgccg ccttcaagta ctttgacacc
    11401 accatcgacc ggaagaggta caccagcacc aaagaggtgc tggacgccac cctgatccac
    11461 cagagcatca ccggcctgta cgagacacgg atcgacctgt ctcagctggg aggcgacgcc
    11521 tatccctatg acgtgcccga ttatgccagc ctgggcagcg gctcccccaa gaaaaaacgc
    11581 aaggtggaag atcctaagaa aaagcggaaa gtggacggca ttggtagtgg gagcaacgga
    11641 tgactcgagc tagttaaaag ttttgttact ttatagaaga aattttgagt ttttgttttt
    11701 ttttaataaa taaataaaca taaataaatt gtttgttgaa tttattatta gtatgtaagt
    11761 gtaaatataa taaaacttaa tatctattca aattaataaa taaacctcga tatacagacc
    11821 gataaaacac atgcgtcaat tttacgcatg attatcttta acgtacgtca caatatgatt
    11881 atctttctag ggttaatcta gtatacgcgt atgcggccgc ttaattaatc cagcttttgt
    11941 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg
    12001 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
    12061 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
    12121 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
    12181 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
    12241 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
    12301 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
    12361 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
    12421 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
    12481 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
    12541 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
    12601 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
    12661 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
    12721 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
    12781 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
    12841 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
    12901 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
    12961 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
    13021 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
    13081 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
    13141 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
    13201 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
    13261 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
    13321 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
    13381 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
    13441 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
    13501 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
    13561 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
    13621 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
    13681 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
    13741 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
    13801 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
    13861 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
    13921 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
    13981 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
    14041 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
    14101 gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa
    14161 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
    14221 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
    14281 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
    14341 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
    14401 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
    14461 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
    14521 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
    14581 agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
    14641 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
    14701 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat
    14761 acgactcact atagggcgaa ttggggcgcg ccattctaga ttaaccctag aaagatagtc
    14821 tgcgtaaaat tgacgcatgc attcttgaaa tattgctctc tctttctaaa tagcgcgaat
    14881 ccgtcgctgt gcatttagga catctcagtc gccgcttgga gctcccgtga ggcgtgcttg
    14941 tcaatgcggt aagtgtcact gattttgaac tataacgacc gcgtgagtca aaatgacgca
    15001 tgattatctt ttacgtgact tttaagattt aactcatacg ataattatat tgttatttca
    15061 tgttctactt acgtgataac ttattatata tatattttct tgttatagat atcaactaga
    15121 atgctagcat gggcccagat ct
//
