LOCUS       sequence-201653-       14728 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Sp dCas9 and GA-inducible KRAB for NOR gate/diametric 
            regulation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14728)
  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 14728)
  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: pSLQ2822 pPB: CAG-GID1-KRAB-IRES-Puro-WPRE
            PGK...
FEATURES             Location/Qualifiers
     source          1..14728
                     /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             2471..2656
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEP"
     misc_feature    join(2674..3247,3248..3250,3251..3259)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2841..2858)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3068..3087
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3281..3880
                     /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(3281..3300)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3777..3797
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    3918..4506
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3971..3991)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4389..4400)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        4527..5026
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4562..4585)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4959..4978
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             5068..5085
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5359..6057
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     regulatory      6073..6082
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6079..6105,6106..6108,6109..8595,8596..8598,
                     8599..10182)
                     /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             6079..10182
                     /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             10186..10212
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             10192..10218
                     /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             10231..10251
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10258..10278
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     repeat_region   11416..11478
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(11529..11549)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(11529..11547)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(11568..11584)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11568..11584)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11581..11603)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11592..11608
                     /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(11616..11622,11623..11640,11641..11646))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11661..11682
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11799..11816)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11970..12558)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12050..12069)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12729..13520,13521..13589))
                     /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     13352..13371
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13590..13694)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(13720..14175)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(13857..13878)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     14069..14088
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     14302..14324
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     14316..14333
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     14317..14333
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     14343..14362
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        14343..14361
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   14391..14425
                     /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 ctagtggcgg tggcggaggg atggatgcta agtcactaac
     2461 tgcctggtcc cggacactgg tgaccttcaa ggatgtattt gtggacttca ccagggagga
     2521 gtggaagctg ctggacactg ctcagcagat cgtgtacaga aatgtgatgc tggagaacta
     2581 taagaacctg gtttccttgg gttatcagct tactaagcca gatgtgatcc tccggttgga
     2641 gaagggagaa gagccctaga tttaaataat tcgcccctct ccctcccccc cccctaacgt
     2701 tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac
     2761 catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag
     2821 cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa
     2881 ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag
     2941 gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga
     3001 tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag
     3061 agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc
     3121 ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacgtgtg tttagtcgag
     3181 gttaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga
     3241 tgataatatg gccacaaccg gatccggagc taccatgacc atgaccgagt acaagcccac
     3301 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt
     3361 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac
     3421 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     3481 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     3541 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     3601 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     3661 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     3721 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     3781 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     3841 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga gtttaaacca tgcaagcttg
     3901 atatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     3961 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     4021 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     4081 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     4141 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     4201 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     4261 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     4321 cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     4381 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     4441 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     4501 ccccgcatcc tgcaggtaat tctaccgggt aggggaggcg cttttcccaa ggcagtctgg
     4561 agcatgcgct ttagcagccc cgctgggcac ttggcgctac acaagtggcc tctggcctcg
     4621 cacacattcc acatccaccg gtaggcgcca accggctccg ttctttggtg gccccttcgc
     4681 gccaccttct actcctcccc tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg
     4741 caggacgtga caaatggaag tagcacgtct cactaggctc gtgcagatgg acagcaccgc
     4801 tgagcaatgg aagcgggtag gcctttgggg cagcggccaa tagcagcttt gctccttcgc
     4861 tttctgggct cagaggctgg gaaggggtgg gtccgggggc gggctcaggg gcgggctcag
     4921 gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg
     4981 cacgtctgcc gcgctgttct cctcttcctc atctccgggc ctttcgacct gcagcccaag
     5041 cttgtcgacg ccaccatgaa gagagatcat catcatcatc atcatcaaga taagaagact
     5101 atgatgatga atgaagaaga cgacggtaac ggcatggatg agcttctagc tgttcttggt
     5161 tacaaggtta ggtcatccga aatggctgat gttgctcaga aactcgagca gcttgaagtt
     5221 atgatgtcta atgttcaaga agacgatctt tctcaactcg ctactgagac tgttcactat
     5281 aatccggcgg agctttacac gtggcttgat tctatgctca ccgaccttaa tcatatgtca
     5341 agcgctgcag gtgctaccat gagcgagctg attaaggaga acatgcacat gaagctgtac
     5401 atggagggca ccgtggacaa ccatcacttc aagtgcacat ccgagggcga aggcaagccc
     5461 tacgagggca cccagaccat gagaatcaag gtggtcgagg gcggccctct ccccttcgcc
     5521 ttcgacatcc tggctactag cttcctctac ggcagcaaga ccttcatcaa ccacacccag
     5581 ggcatccccg acttcttcaa gcagtccttc cctgagggct tcacatggga gagagtcacc
     5641 acatacgaag acgggggcgt gctgaccgct acccaggaca ccagcctcca ggacggctgc
     5701 ctcatctaca acgtcaagat cagaggggtg aacttcacat ccaacggccc tgtgatgcag
     5761 aagaaaacac tcggctggga ggccttcacc gagacgctgt accccgctga cggcggcctg
     5821 gaaggcagaa acgacatggc cctgaagctc gtgggcggga gccatctgat cgcaaacatc
     5881 aagaccacat atagatccaa gaaacccgct aagaacctca agatgcctgg cgtctactat
     5941 gtggactaca gactggaaag aatcaaggag gccaacaacg agacctacgt cgagcagcac
     6001 gaggtggcag tggccagata ctgcgacctc cctagcaaac tggggcacaa gcttaattca
     6061 agcgctgcag gtgccaccat ggacaagaag tacagcatcg gcctggccat cggcaccaac
     6121 tctgtgggct gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg
     6181 ctgggcaaca ccgaccggca cagcatcaag aagaacctga tcggcgccct gctgttcgac
     6241 agcggagaaa cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga
     6301 cggaagaacc ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac
     6361 gacagcttct tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag
     6421 cggcacccca tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc
     6481 atctaccacc tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc
     6541 tatctggccc tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg
     6601 aaccccgaca acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag
     6661 ctgttcgagg aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc
     6721 agactgagca agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag
     6781 aatggcctgt tcggcaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc
     6841 aacttcgacc tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac
     6901 ctggacaacc tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag
     6961 aacctgtccg acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag
     7021 gcccccctga gcgcctctat gatcaagaga tacgacgagc accaccagga cctgaccctg
     7081 ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag
     7141 agcaagaacg gctacgccgg ctacatcgat ggcggagcca gccaggaaga gttctacaag
     7201 ttcatcaagc ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac
     7261 agagaggacc tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc ccaccagatc
     7321 cacctgggag agctgcacgc cattctgcgg cggcaggaag atttttaccc attcctgaag
     7381 gacaaccggg aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct
     7441 ctggccaggg gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc
     7501 ccctggaact tcgaggaagt ggtggacaag ggcgccagcg cccagagctt catcgagcgg
     7561 atgaccaact tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg
     7621 tacgagtact tcaccgtgta caacgagctg accaaagtga aatacgtgac cgagggaatg
     7681 agaaagcccg ccttcctgag cggcgagcag aaaaaagcca tcgtggacct gctgttcaag
     7741 accaaccgga aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc
     7801 ttcgactccg tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac
     7861 cacgatctgc tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac
     7921 attctggaag atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa
     7981 cggctgaaaa cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggcgg
     8041 agatacaccg gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag
     8101 tccggcaaga caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg
     8161 cagctgatcc acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc
     8221 ggccagggcg atagcctgca cgagcacatt gccaatctgg ccggcagccc cgccattaag
     8281 aagggcatcc tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat gggccggcac
     8341 aagcccgaga acatcgtgat cgaaatggcc agagagaacc agaccaccca gaagggacag
     8401 aagaacagcc gcgagagaat gaagcggatc gaagagggca tcaaagagct gggcagccag
     8461 atcctgaaag aacaccccgt ggaaaacacc cagctgcaga acgagaagct gtacctgtac
     8521 tacctgcaga atgggcggga tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc
     8581 gactacgatg tggacgctat cgtgcctcag agctttctga aggacgactc catcgataac
     8641 aaagtgctga ctcggagcga caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag
     8701 gtcgtgaaga agatgaagaa ctactggcgc cagctgctga atgccaagct gattacccag
     8761 aggaagttcg acaatctgac caaggccgag agaggcggcc tgagcgaact ggataaggcc
     8821 ggcttcatca agagacagct ggtggaaacc cggcagatca caaagcacgt ggcacagatc
     8881 ctggactccc ggatgaacac taagtacgac gagaacgaca aactgatccg ggaagtgaaa
     8941 gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga aggatttcca gttttacaaa
     9001 gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga
     9061 accgccctga tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag
     9121 gtgtacgacg tgcggaagat gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc
     9181 aagtacttct tctacagcaa catcatgaac tttttcaaga ccgagattac cctggccaac
     9241 ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg aaacaggcga gatcgtgtgg
     9301 gataagggcc gggactttgc caccgtgcgg aaagtgctgt ctatgcccca agtgaatatc
     9361 gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag agtctatcct gcccaagagg
     9421 aacagcgaca agctgatcgc cagaaagaag gactgggacc ctaagaagta cggcggcttc
     9481 gacagcccca ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc
     9541 aagaaactga agagtgtgaa agagctgctg gggatcacca tcatggaaag aagcagcttc
     9601 gagaagaatc ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg
     9661 atcatcaagc tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg
     9721 gcctctgccg gcgaactgca gaagggaaac gaactggccc tgccctccaa atatgtgaac
     9781 ttcctgtacc tggccagcca ctatgagaag ctgaagggct cccccgagga taatgagcag
     9841 aaacagctgt ttgtggaaca gcacaaacac tacctggacg agatcatcga gcagatcagc
     9901 gagttctcca agagagtgat cctggccgac gctaatctgg acaaggtgct gagcgcctac
     9961 aacaagcaca gagacaagcc tatcagagag caggccgaga atatcatcca cctgtttacc
    10021 ctgaccaatc tgggagcccc tgccgccttc aagtactttg acaccaccat cgaccggaag
    10081 aggtacacca gcaccaaaga ggtgctggac gccaccctga tccaccagag catcaccggc
    10141 ctgtacgaga cacggatcga cctgtctcag ctgggaggcg acgcctatcc ctatgacgtg
    10201 cccgattatg ccagcctggg cagcggctcc cccaagaaaa aacgcaaggt ggaagatcct
    10261 aagaaaaagc ggaaagtgga cggcattggt agtgggagca acggaggtgg aggctctgga
    10321 cctaggacgc gtgtgccttt gtatggtttt acttcgattt gtggaagaag acctgagatg
    10381 gaagctgctg tttcgactat accaagattc cttcaatctt cctctggttc gatgttagat
    10441 ggtcggtttg atcctcaatc cgccgctcat ttcttcggtg tttacgacgg ccatggcggt
    10501 tctcaggtag cgaactattg tagagagagg atgcatttgg ctttggcgga ggagatagct
    10561 aaggagaaac cgatgctctg cgatggtgat acgtggctgg agaagtggaa gaaagctctt
    10621 ttcaactcgt tcctgagagt tgactcggag attgagtcag ttgcgccgga gacggttggg
    10681 tcaacgtcgg tggttgccgt tgttttcccg tctcacatct tcgtcgctaa ctgcggtgac
    10741 tctagagccg ttctttgccg cggcaaaact gcacttccat tatccgttga ccataaaccg
    10801 gatagagaag atgaagctgc gaggattgaa gccgcaggag ggaaagtgat tcagtggaat
    10861 ggagctcgtg ttttcggtgt tctcgccatg tcgagatcca ttggcgatag atacttgaaa
    10921 ccatccatca ttcctgatcc ggaagtgacg gctgtgaaga gagtaaaaga agatgattgt
    10981 ctgattttgg cgagtgacgg ggtttgggat gtaatgacgg atgaagaagc gtgtgagatg
    11041 gcaaggaagc ggattctctt gtggcacaag aaaaacgcgg tggctgggga tgcatcgttg
    11101 ctcgcggatg agcggagaaa ggaagggaaa gatcctgcgg cgatgtccgc ggctgagtat
    11161 ttgtcaaagc tggcgataca gagaggaagc aaagacaaca taagtgtggt ggtggttgat
    11221 ttgaagtgac tcgagctagt taaaagtttt gttactttat agaagaaatt ttgagttttt
    11281 gttttttttt aataaataaa taaacataaa taaattgttt gttgaattta ttattagtat
    11341 gtaagtgtaa atataataaa acttaatatc tattcaaatt aataaataaa cctcgatata
    11401 cagaccgata aaacacatgc gtcaatttta cgcatgatta tctttaacgt acgtcacaat
    11461 atgattatct ttctagggtt aatctagtat acgcgtatgc ggccgcttaa ttaatccagc
    11521 ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt
    11581 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
    11641 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
    11701 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
    11761 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
    11821 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
    11881 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
    11941 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
    12001 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
    12061 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
    12121 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
    12181 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
    12241 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
    12301 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
    12361 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
    12421 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
    12481 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
    12541 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
    12601 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
    12661 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
    12721 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
    12781 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
    12841 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
    12901 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
    12961 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
    13021 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
    13081 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
    13141 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
    13201 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
    13261 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
    13321 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
    13381 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
    13441 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
    13501 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
    13561 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
    13621 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
    13681 ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt
    13741 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
    13801 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
    13861 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
    13921 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
    13981 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
    14041 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
    14101 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
    14161 cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
    14221 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
    14281 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg
    14341 cgtaatacga ctcactatag ggcgaattgg ggcgcgccat tctagattaa ccctagaaag
    14401 atagtctgcg taaaattgac gcatgcattc ttgaaatatt gctctctctt tctaaatagc
    14461 gcgaatccgt cgctgtgcat ttaggacatc tcagtcgccg cttggagctc ccgtgaggcg
    14521 tgcttgtcaa tgcggtaagt gtcactgatt ttgaactata acgaccgcgt gagtcaaaat
    14581 gacgcatgat tatcttttac gtgactttta agatttaact catacgataa ttatattgtt
    14641 atttcatgtt ctacttacgt gataacttat tatatatata ttttcttgtt atagatatca
    14701 actagaatgc tagcatgggc ccagatct
//
