LOCUS       sequence-160811-       15416 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ABA-inducible VPR-Sp dCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15416)
  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 15416)
  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: pSLQ2817 pPB: CAG-PYL1-VPR-IRES-Puro-WPRE-SV40...
FEATURES             Location/Qualifiers
     source          1..15416
                     /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             1940..2089
                     /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             2096..2116
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2546..2917
                     /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             2936..3505
                     /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(3523..4096,4097..4099,4100..4108)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3690..3707)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3917..3936
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4130..4729
                     /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(4130..4149)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4626..4646
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4767..5355
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4820..4840)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5238..5249)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5394..5413)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5448..5467
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        5509..6008
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5544..5567)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     5941..5960
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6968..7666
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     regulatory      7682..7691
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(7688..7714,7715..7717,7718..10204,10205..10207,
                     10208..11791)
                     /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             7688..11791
                     /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             11795..11821
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             11801..11827
                     /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             11840..11860
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11867..11887
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     repeat_region   12104..12166
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(12217..12237)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(12217..12235)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(12256..12272)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12256..12272)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12269..12291)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12280..12296
                     /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(12304..12310,12311..12328,12329..12334))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12349..12370
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12487..12504)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12658..13246)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12738..12757)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13417..14208,14209..14277))
                     /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     14040..14059
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14278..14382)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(14408..14863)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(14545..14566)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     14757..14776
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     14990..15012
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     15004..15021
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     15005..15021
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     15031..15050
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        15031..15049
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   15079..15113
                     /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 accatgggtg ggggcgcgcc
     1381 aactcaagac gaattcaccc aactctccca atcaatcgcc gagttccaca cgtaccaact
     1441 cggtaacggc cgttgctcat ctctcctagc tcagcgaatc cacgcgccgc cggaaacagt
     1501 atggtccgtg gtgagacgtt tcgataggcc acagatttac aaacacttca tcaaaagctg
     1561 taacgtgagt gaagatttcg agatgcgagt gggatgcacg cgcgacgtga acgtgataag
     1621 tggattaccg gcgaatacgt ctcgagagag attagatctg ttggacgatg atcggagagt
     1681 gactgggttt agtataaccg gtggtgaaca taggctgagg aattataaat cggttacgac
     1741 ggttcataga tttgagaaag aagaagaaga agaaaggatc tggaccgttg ttttggaatc
     1801 ttatgttgtt gatgtaccgg aaggtaattc ggaggaagat acgagattgt ttgctgatac
     1861 ggttattaga ttgaatcttc agaaacttgc ttcgatcact gaagctatga acgaattctc
     1921 aagcgctgca ggtactagtg atgctttaga cgattttgac ttagatatgc ttggttcaga
     1981 cgcgttagac gacttcgacc tagacatgtt aggctcagat gcattggacg acttcgattt
     2041 agatatgttg ggctccgatg ccctagatga ctttgatcta gatatgctag gtagtcccaa
     2101 aaagaagagg aaagtgggat cccagtatct gcccgacaca gatgatagac accgaatcga
     2161 agagaaacgc aagcgaacgt atgaaacctt caaatcgatc atgaagaaat cgcccttctc
     2221 gggtccgacc gatcccaggc ccccaccgag aaggattgcg gtcccgtccc gctcgtcggc
     2281 cagcgtgccg aagcctgcgc cgcagcccta ccccttcacg tcgagcctga gcacaatcaa
     2341 ttatgacgag ttcccgacga tggtgttccc ctcgggacaa atctcacaag cctcggcgct
     2401 cgcaccagcg cctccccaag tccttccgca agcgcctgcc ccagcgcctg caccggcaat
     2461 ggtgtccgcc ctcgcacagg cccctgcgcc cgtccccgtg ctcgcgcctg gaccgcccca
     2521 ggcggtcgct ccaccggctc cgaagccgac gcaggccgga gagggaacac tctccgaagc
     2581 acttcttcaa ctccagtttg atgacgagga tcttggagca ctccttggaa actcgacaga
     2641 ccctgcggtg tttaccgacc tcgcgtcagt agataactcc gaatttcagc agcttttgaa
     2701 ccagggtatc ccggtcgcgc cacatacaac ggagcccatg ttgatggaat accccgaagc
     2761 aatcacgaga cttgtgacgg gagcgcagcg gcctcccgat cccgcacccg cacctttggg
     2821 ggcacctggc ctccctaacg gacttttgag cggcgacgag gatttctcct ccatcgccga
     2881 tatggatttc tcagccttgc tgtcacagat ttccagcggc tctggcagcg gcagccggga
     2941 ttccagggaa gggatgtttt tgccgaagcc tgaggccggc tccgctatta gtgacgtgtt
     3001 tgagggccgc gaggtgtgcc agccaaaacg aatccggcca tttcatcctc caggaagtcc
     3061 atgggccaac cgcccactcc ccgccagcct cgcaccaaca ccaaccggtc cagtacatga
     3121 gccagtcggg tcactgaccc cggcaccagt ccctcagcca ctggatccag cgcccgcagt
     3181 gactcccgag gccagtcacc tgttggagga tcccgatgaa gagacgagcc aggctgtcaa
     3241 agcccttcgg gagatggccg atactgtgat tccccagaag gaagaggctg caatctgtgg
     3301 ccaaatggac ctttcccatc cgcccccaag gggccatctg gatgagctga caaccacact
     3361 tgagtccatg accgaggatc tgaacctgga ctcacccctg accccggaat tgaacgagat
     3421 tctggatacc ttcctgaacg acgagtgcct cttgcatgcc atgcatatca gcacaggact
     3481 gtccatcttc gacacatctc tgttttgaat ttaaataatt cgcccctctc cctccccccc
     3541 ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt
     3601 attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt
     3661 cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa
     3721 tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac
     3781 cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg
     3841 tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt
     3901 tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca
     3961 gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacgtgtgt
     4021 ttagtcgagg ttaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga
     4081 aaaacacgat gataatatgg ccacaaccgg atccggagct accatgacca tgaccgagta
     4141 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc
     4201 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga
     4261 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
     4321 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
     4381 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
     4441 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
     4501 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
     4561 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
     4621 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
     4681 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgag tttaaaccat
     4741 gcaagcttga tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat
     4801 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     4861 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     4921 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     4981 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     5041 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     5101 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     5161 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     5221 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     5281 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     5341 gggccgcctc cccgcatcct tgtttattgc agcttataat ggttacaaat aaagcaatag
     5401 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5461 actcatcaat gtatcttatc atgtctggat cctgcaggta attctaccgg gtaggggagg
     5521 cgcttttccc aaggcagtct ggagcatgcg ctttagcagc cccgctgggc acttggcgct
     5581 acacaagtgg cctctggcct cgcacacatt ccacatccac cggtaggcgc caaccggctc
     5641 cgttctttgg tggccccttc gcgccacctt ctactcctcc cctagtcagg aagttccccc
     5701 ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga agtagcacgt ctcactaggc
     5761 tcgtgcagat ggacagcacc gctgagcaat ggaagcgggt aggcctttgg ggcagcggcc
     5821 aatagcagct ttgctccttc gctttctggg ctcagaggct gggaaggggt gggtccgggg
     5881 gcgggctcag gggcgggctc aggggcgggg cgggcgcccg aaggtcctcc ggaggcccgg
     5941 cattctgcac gcttcaaaag cgcacgtctg ccgcgctgtt ctcctcttcc tcatctccgg
     6001 gcctttcgac ctgcagccca agcttgtcga cgccaccatg acgcgtgtgc ctttgtatgg
     6061 ttttacttcg atttgtggaa gaagacctga gatggaagct gctgtttcga ctataccaag
     6121 attccttcaa tcttcctctg gttcgatgtt agatggtcgg tttgatcctc aatccgccgc
     6181 tcatttcttc ggtgtttacg acggccatgg cggttctcag gtagcgaact attgtagaga
     6241 gaggatgcat ttggctttgg cggaggagat agctaaggag aaaccgatgc tctgcgatgg
     6301 tgatacgtgg ctggagaagt ggaagaaagc tcttttcaac tcgttcctga gagttgactc
     6361 ggagattgag tcagttgcgc cggagacggt tgggtcaacg tcggtggttg ccgttgtttt
     6421 cccgtctcac atcttcgtcg ctaactgcgg tgactctaga gccgttcttt gccgcggcaa
     6481 aactgcactt ccattatccg ttgaccataa accggataga gaagatgaag ctgcgaggat
     6541 tgaagccgca ggagggaaag tgattcagtg gaatggagct cgtgttttcg gtgttctcgc
     6601 catgtcgaga tccattggcg atagatactt gaaaccatcc atcattcctg atccggaagt
     6661 gacggctgtg aagagagtaa aagaagatga ttgtctgatt ttggcgagtg acggggtttg
     6721 ggatgtaatg acggatgaag aagcgtgtga gatggcaagg aagcggattc tcttgtggca
     6781 caagaaaaac gcggtggctg gggatgcatc gttgctcgcg gatgagcgga gaaaggaagg
     6841 gaaagatcct gcggcgatgt ccgcggctga gtatttgtca aagctggcga tacagagagg
     6901 aagcaaagac aacataagtg tggtggtggt tgatttgaag catatgtcaa gcgctgcagg
     6961 tgctaccatg agcgagctga ttaaggagaa catgcacatg aagctgtaca tggagggcac
     7021 cgtggacaac catcacttca agtgcacatc cgagggcgaa ggcaagccct acgagggcac
     7081 ccagaccatg agaatcaagg tggtcgaggg cggccctctc cccttcgcct tcgacatcct
     7141 ggctactagc ttcctctacg gcagcaagac cttcatcaac cacacccagg gcatccccga
     7201 cttcttcaag cagtccttcc ctgagggctt cacatgggag agagtcacca catacgaaga
     7261 cgggggcgtg ctgaccgcta cccaggacac cagcctccag gacggctgcc tcatctacaa
     7321 cgtcaagatc agaggggtga acttcacatc caacggccct gtgatgcaga agaaaacact
     7381 cggctgggag gccttcaccg agacgctgta ccccgctgac ggcggcctgg aaggcagaaa
     7441 cgacatggcc ctgaagctcg tgggcgggag ccatctgatc gcaaacatca agaccacata
     7501 tagatccaag aaacccgcta agaacctcaa gatgcctggc gtctactatg tggactacag
     7561 actggaaaga atcaaggagg ccaacaacga gacctacgtc gagcagcacg aggtggcagt
     7621 ggccagatac tgcgacctcc ctagcaaact ggggcacaag cttaattcaa gcgctgcagg
     7681 tgccaccatg gacaagaagt acagcatcgg cctggccatc ggcaccaact ctgtgggctg
     7741 ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac
     7801 cgaccggcac agcatcaaga agaacctgat cggcgccctg ctgttcgaca gcggagaaac
     7861 agccgaggcc acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg
     7921 gatctgctat ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt
     7981 ccacagactg gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat
     8041 cttcggcaac atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct
     8101 gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct
     8161 ggcccacatg atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa
     8221 cagcgacgtg gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga
     8281 aaaccccatc aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa
     8341 gagcagacgg ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt
     8401 cggcaacctg attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct
     8461 ggccgaggat gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct
     8521 gctggcccag atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga
     8581 cgccatcctg ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag
     8641 cgcctctatg atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct
     8701 cgtgcggcag cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg
     8761 ctacgccggc tacatcgatg gcggagccag ccaggaagag ttctacaagt tcatcaagcc
     8821 catcctggaa aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct
     8881 gctgcggaag cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga
     8941 gctgcacgcc attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga
     9001 aaagatcgag aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg
     9061 aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt
     9121 cgaggaagtg gtggacaagg gcgccagcgc ccagagcttc atcgagcgga tgaccaactt
     9181 cgataagaac ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt
     9241 caccgtgtac aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc
     9301 cttcctgagc ggcgagcaga aaaaagccat cgtggacctg ctgttcaaga ccaaccggaa
     9361 agtgaccgtg aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt
     9421 ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct
     9481 gaaaattatc aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga
     9541 tatcgtgctg accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac
     9601 ctatgcccac ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg
     9661 ctggggcagg ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac
     9721 aatcctggat ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca
     9781 cgacgacagc ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga
     9841 tagcctgcac gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct
     9901 gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa
     9961 catcgtgatc gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg
    10021 cgagagaatg aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga
    10081 acaccccgtg gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa
    10141 tgggcgggat atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt
    10201 ggacgctatc gtgcctcaga gctttctgaa ggacgactcc atcgataaca aagtgctgac
    10261 tcggagcgac aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa
    10321 gatgaagaac tactggcgcc agctgctgaa tgccaagctg attacccaga ggaagttcga
    10381 caatctgacc aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa
    10441 gagacagctg gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg
    10501 gatgaacact aagtacgacg agaacgacaa actgatccgg gaagtgaaag tgatcaccct
    10561 gaagtccaag ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat
    10621 caacaactac caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat
    10681 caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt
    10741 gcggaagatg atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt
    10801 ctacagcaac atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg
    10861 gaagcggcct ctgatcgaga caaacggcga aacaggcgag atcgtgtggg ataagggccg
    10921 ggactttgcc accgtgcgga aagtgctgtc tatgccccaa gtgaatatcg tgaaaaagac
    10981 cgaggtgcag acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgacaa
    11041 gctgatcgcc agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac
    11101 cgtggcctat tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa
    11161 gagtgtgaaa gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc
    11221 catcgacttt ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct
    11281 gcctaagtac tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg
    11341 cgaactgcag aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct
    11401 ggccagccac tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt
    11461 tgtggaacag cacaaacact acctggacga gatcatcgag cagatcagcg agttctccaa
    11521 gagagtgatc ctggccgacg ctaatctgga caaggtgctg agcgcctaca acaagcacag
    11581 agacaagcct atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct
    11641 gggagcccct gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag
    11701 caccaaagag gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac
    11761 acggatcgac ctgtctcagc tgggaggcga cgcctatccc tatgacgtgc ccgattatgc
    11821 cagcctgggc agcggctccc ccaagaaaaa acgcaaggtg gaagatccta agaaaaagcg
    11881 gaaagtggac ggcattggta gtgggagcaa cggatgactc gagctagtta aaagttttgt
    11941 tactttatag aagaaatttt gagtttttgt ttttttttaa taaataaata aacataaata
    12001 aattgtttgt tgaatttatt attagtatgt aagtgtaaat ataataaaac ttaatatcta
    12061 ttcaaattaa taaataaacc tcgatataca gaccgataaa acacatgcgt caattttacg
    12121 catgattatc tttaacgtac gtcacaatat gattatcttt ctagggttaa tctagtatac
    12181 gcgtatgcgg ccgcttaatt aatccagctt ttgttccctt tagtgagggt taattgcgcg
    12241 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
    12301 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
    12361 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
    12421 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
    12481 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
    12541 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
    12601 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
    12661 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
    12721 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
    12781 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
    12841 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
    12901 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
    12961 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
    13021 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
    13081 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
    13141 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
    13201 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
    13261 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
    13321 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
    13381 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
    13441 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
    13501 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
    13561 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
    13621 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
    13681 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
    13741 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
    13801 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
    13861 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
    13921 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
    13981 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
    14041 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
    14101 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
    14161 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
    14221 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
    14281 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
    14341 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
    14401 gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
    14461 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
    14521 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
    14581 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
    14641 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
    14701 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
    14761 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
    14821 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg
    14881 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
    14941 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
    15001 cgttgtaaaa cgacggccag tgagcgcgcg taatacgact cactataggg cgaattgggg
    15061 cgcgccattc tagattaacc ctagaaagat agtctgcgta aaattgacgc atgcattctt
    15121 gaaatattgc tctctctttc taaatagcgc gaatccgtcg ctgtgcattt aggacatctc
    15181 agtcgccgct tggagctccc gtgaggcgtg cttgtcaatg cggtaagtgt cactgatttt
    15241 gaactataac gaccgcgtga gtcaaaatga cgcatgatta tcttttacgt gacttttaag
    15301 atttaactca tacgataatt atattgttat ttcatgttct acttacgtga taacttatta
    15361 tatatatatt ttcttgttat agatatcaac tagaatgcta gcatgggccc agatct
//
