LOCUS       sequence-205810-       13779 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses Sp dCas9 and GA-inducible KRAB for NOR gate/diametric 
            regulation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 13779)
  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 13779)
  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..13779
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     272..294
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     356..375
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      403..412
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1516..1701
                     /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(1719..2292,2293..2295,2296..2304)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1886..1903)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2113..2132
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2326..2925
                     /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(2326..2345)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2822..2842
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    2963..3551
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3016..3036)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3434..3445)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        3572..4071
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3607..3630)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4004..4023
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             4113..4130
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4404..5102
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     regulatory      5118..5127
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5124..5150,5151..5153,5154..7640,7641..7643,
                     7644..9227)
                     /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             5124..9227
                     /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             9231..9257
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             9237..9263
                     /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             9276..9296
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9303..9323
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     repeat_region   10461..10523
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(10574..10594)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(10574..10592)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(10613..10629)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10613..10629)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10626..10648)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10637..10653
                     /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(10661..10667,10668..10685,10686..10691))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10706..10727
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10844..10861)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11015..11603)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11095..11114)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11774..12565,12566..12634))
                     /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     12397..12416
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12635..12739)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(12765..13220)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12902..12923)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     13114..13133
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     13347..13369
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     13361..13378
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     13362..13378
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     13388..13407
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        13388..13406
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   13436..13470
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc ccagggactt
       61 cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac cccctctagc
      121 gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt
      181 gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc gcggggggac
      241 ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt gaccggcggc
      301 tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac
      361 gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcgccaccat ggctgcgagc
      421 gatgaagtta atcttattga gagcagaaca gtggttcctc tcaatacatg ggttttaata
      481 tccaacttca aagtagccta caatatcctt cgtcgccctg atggaacctt taaccgacac
      541 ttagctgagt atctagaccg taaagtcact gcaaacgcca atccggttga tggggttttc
      601 tcgttcgatg tcttgattga tcgcaggatc aatcttctaa gcagagtcta tagaccagct
      661 tatgcagatc aagagcaacc tcctagtatt ttagatctcg agaagcctgt tgatggcgac
      721 attgtccctg ttatattgtt cttccatgga ggtagctttg ctcattcttc tgcaaacagt
      781 gccatctacg atactctttg tcgcaggctt gttggtttgt gcaagtgtgt tgttgtctct
      841 gtgaattatc ggcgtgcacc agagaatcca tacccttgtg cttatgatga tggttggatt
      901 gctcttaatt gggttaactc gagatcttgg cttaaatcca agaaagactc aaaggtccat
      961 attttcttgg ctggtgatag ctctggaggt aacatcgcgc ataatgtggc tttaagagcg
     1021 ggtgaatcgg gaatcgatgt tttggggaac attctgctga atcctatgtt tggtgggaat
     1081 gagagaacgg agtctgagaa aagtttggat gggaaatact ttgtgacggt tagagaccgc
     1141 gattggtact ggaaagcgtt tttacccgag ggagaagata gagagcatcc agcgtgtaat
     1201 ccgtttagcc cgagagggaa aagcttagaa ggagtgagtt tccccaagag tcttgtggtt
     1261 gtcgcgggtt tggatttgat tagagattgg cagttggcat acgcggaagg gctcaagaaa
     1321 gcgggtcaag aggttaagct tatgcattta gagaaagcaa ctgttgggtt ttacctcttg
     1381 cctaataaca atcatttcca taatgttatg gatgagattt cggcgtttgt aaacgcggaa
     1441 tgtgaattct caagcgctgc aggtactagt ggcggtggcg gagggatgga tgctaagtca
     1501 ctaactgcct ggtcccggac actggtgacc ttcaaggatg tatttgtgga cttcaccagg
     1561 gaggagtgga agctgctgga cactgctcag cagatcgtgt acagaaatgt gatgctggag
     1621 aactataaga acctggtttc cttgggttat cagcttacta agccagatgt gatcctccgg
     1681 ttggagaagg gagaagagcc ctagatttaa ataattcgcc cctctccctc ccccccccct
     1741 aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt
     1801 tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg
     1861 acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc
     1921 gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt
     1981 tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta
     2041 taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg
     2101 gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag
     2161 gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac gtgtgtttag
     2221 tcgaggttaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa
     2281 cacgatgata atatggccac aaccggatcc ggagctacca tgaccatgac cgagtacaag
     2341 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     2401 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     2461 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     2521 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     2581 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     2641 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     2701 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     2761 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     2821 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     2881 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgagttta aaccatgcaa
     2941 gcttgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
     3001 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     3061 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     3121 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     3181 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     3241 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     3301 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     3361 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     3421 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     3481 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     3541 cgcctccccg catcctgcag gtaattctac cgggtagggg aggcgctttt cccaaggcag
     3601 tctggagcat gcgctttagc agccccgctg ggcacttggc gctacacaag tggcctctgg
     3661 cctcgcacac attccacatc caccggtagg cgccaaccgg ctccgttctt tggtggcccc
     3721 ttcgcgccac cttctactcc tcccctagtc aggaagttcc cccccgcccc gcagctcgcg
     3781 tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta ggctcgtgca gatggacagc
     3841 accgctgagc aatggaagcg ggtaggcctt tggggcagcg gccaatagca gctttgctcc
     3901 ttcgctttct gggctcagag gctgggaagg ggtgggtccg ggggcgggct caggggcggg
     3961 ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa
     4021 aagcgcacgt ctgccgcgct gttctcctct tcctcatctc cgggcctttc gacctgcagc
     4081 ccaagcttgt cgacgccacc atgaagagag atcatcatca tcatcatcat caagataaga
     4141 agactatgat gatgaatgaa gaagacgacg gtaacggcat ggatgagctt ctagctgttc
     4201 ttggttacaa ggttaggtca tccgaaatgg ctgatgttgc tcagaaactc gagcagcttg
     4261 aagttatgat gtctaatgtt caagaagacg atctttctca actcgctact gagactgttc
     4321 actataatcc ggcggagctt tacacgtggc ttgattctat gctcaccgac cttaatcata
     4381 tgtcaagcgc tgcaggtgct accatgagcg agctgattaa ggagaacatg cacatgaagc
     4441 tgtacatgga gggcaccgtg gacaaccatc acttcaagtg cacatccgag ggcgaaggca
     4501 agccctacga gggcacccag accatgagaa tcaaggtggt cgagggcggc cctctcccct
     4561 tcgccttcga catcctggct actagcttcc tctacggcag caagaccttc atcaaccaca
     4621 cccagggcat ccccgacttc ttcaagcagt ccttccctga gggcttcaca tgggagagag
     4681 tcaccacata cgaagacggg ggcgtgctga ccgctaccca ggacaccagc ctccaggacg
     4741 gctgcctcat ctacaacgtc aagatcagag gggtgaactt cacatccaac ggccctgtga
     4801 tgcagaagaa aacactcggc tgggaggcct tcaccgagac gctgtacccc gctgacggcg
     4861 gcctggaagg cagaaacgac atggccctga agctcgtggg cgggagccat ctgatcgcaa
     4921 acatcaagac cacatataga tccaagaaac ccgctaagaa cctcaagatg cctggcgtct
     4981 actatgtgga ctacagactg gaaagaatca aggaggccaa caacgagacc tacgtcgagc
     5041 agcacgaggt ggcagtggcc agatactgcg acctccctag caaactgggg cacaagctta
     5101 attcaagcgc tgcaggtgcc accatggaca agaagtacag catcggcctg gccatcggca
     5161 ccaactctgt gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc aagaaattca
     5221 aggtgctggg caacaccgac cggcacagca tcaagaagaa cctgatcggc gccctgctgt
     5281 tcgacagcgg agaaacagcc gaggccaccc ggctgaagag aaccgccaga agaagataca
     5341 ccagacggaa gaaccggatc tgctatctgc aagagatctt cagcaacgag atggccaagg
     5401 tggacgacag cttcttccac agactggaag agtccttcct ggtggaagag gataagaagc
     5461 acgagcggca ccccatcttc ggcaacatcg tggacgaggt ggcctaccac gagaagtacc
     5521 ccaccatcta ccacctgaga aagaaactgg tggacagcac cgacaaggcc gacctgcggc
     5581 tgatctatct ggccctggcc cacatgatca agttccgggg ccacttcctg atcgagggcg
     5641 acctgaaccc cgacaacagc gacgtggaca agctgttcat ccagctggtg cagacctaca
     5701 accagctgtt cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag gccatcctgt
     5761 ctgccagact gagcaagagc agacggctgg aaaatctgat cgcccagctg cccggcgaga
     5821 agaagaatgg cctgttcggc aacctgattg ccctgagcct gggcctgacc cccaacttca
     5881 agagcaactt cgacctggcc gaggatgcca aactgcagct gagcaaggac acctacgacg
     5941 acgacctgga caacctgctg gcccagatcg gcgaccagta cgccgacctg tttctggccg
     6001 ccaagaacct gtccgacgcc atcctgctga gcgacatcct gagagtgaac accgagatca
     6061 ccaaggcccc cctgagcgcc tctatgatca agagatacga cgagcaccac caggacctga
     6121 ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa gtacaaagag attttcttcg
     6181 accagagcaa gaacggctac gccggctaca tcgatggcgg agccagccag gaagagttct
     6241 acaagttcat caagcccatc ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc
     6301 tgaacagaga ggacctgctg cggaagcagc ggaccttcga caacggcagc atcccccacc
     6361 agatccacct gggagagctg cacgccattc tgcggcggca ggaagatttt tacccattcc
     6421 tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc tactacgtgg
     6481 gccctctggc caggggaaac agcagattcg cctggatgac cagaaagagc gaggaaacca
     6541 tcaccccctg gaacttcgag gaagtggtgg acaagggcgc cagcgcccag agcttcatcg
     6601 agcggatgac caacttcgat aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc
     6661 tgctgtacga gtacttcacc gtgtacaacg agctgaccaa agtgaaatac gtgaccgagg
     6721 gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa agccatcgtg gacctgctgt
     6781 tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga ggactacttc aagaaaatcg
     6841 agtgcttcga ctccgtggaa atctccggcg tggaagatcg gttcaacgcc tccctgggca
     6901 cataccacga tctgctgaaa attatcaagg acaaggactt cctggacaat gaggaaaacg
     6961 aggacattct ggaagatatc gtgctgaccc tgacactgtt tgaggacaga gagatgatcg
     7021 aggaacggct gaaaacctat gcccacctgt tcgacgacaa agtgatgaag cagctgaagc
     7081 ggcggagata caccggctgg ggcaggctga gccggaagct gatcaacggc atccgggaca
     7141 agcagtccgg caagacaatc ctggatttcc tgaagtccga cggcttcgcc aacagaaact
     7201 tcatgcagct gatccacgac gacagcctga cctttaaaga ggacatccag aaagcccagg
     7261 tgtccggcca gggcgatagc ctgcacgagc acattgccaa tctggccggc agccccgcca
     7321 ttaagaaggg catcctgcag acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc
     7381 ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga gaaccagacc acccagaagg
     7441 gacagaagaa cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa gagctgggca
     7501 gccagatcct gaaagaacac cccgtggaaa acacccagct gcagaacgag aagctgtacc
     7561 tgtactacct gcagaatggg cgggatatgt acgtggacca ggaactggac atcaaccggc
     7621 tgtccgacta cgatgtggac gctatcgtgc ctcagagctt tctgaaggac gactccatcg
     7681 ataacaaagt gctgactcgg agcgacaaga accggggcaa gagcgacaac gtgccctccg
     7741 aagaggtcgt gaagaagatg aagaactact ggcgccagct gctgaatgcc aagctgatta
     7801 cccagaggaa gttcgacaat ctgaccaagg ccgagagagg cggcctgagc gaactggata
     7861 aggccggctt catcaagaga cagctggtgg aaacccggca gatcacaaag cacgtggcac
     7921 agatcctgga ctcccggatg aacactaagt acgacgagaa cgacaaactg atccgggaag
     7981 tgaaagtgat caccctgaag tccaagctgg tgtccgattt ccggaaggat ttccagtttt
     8041 acaaagtgcg cgagatcaac aactaccacc acgcccacga cgcctacctg aacgccgtcg
     8101 tgggaaccgc cctgatcaaa aagtacccta agctggaaag cgagttcgtg tacggcgact
     8161 acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta
     8221 ccgccaagta cttcttctac agcaacatca tgaacttttt caagaccgag attaccctgg
     8281 ccaacggcga gatccggaag cggcctctga tcgagacaaa cggcgaaaca ggcgagatcg
     8341 tgtgggataa gggccgggac tttgccaccg tgcggaaagt gctgtctatg ccccaagtga
     8401 atatcgtgaa aaagaccgag gtgcagacag gcggcttcag caaagagtct atcctgccca
     8461 agaggaacag cgacaagctg atcgccagaa agaaggactg ggaccctaag aagtacggcg
     8521 gcttcgacag ccccaccgtg gcctattctg tgctggtggt ggccaaagtg gaaaagggca
     8581 agtccaagaa actgaagagt gtgaaagagc tgctggggat caccatcatg gaaagaagca
     8641 gcttcgagaa gaatcccatc gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg
     8701 acctgatcat caagctgcct aagtactccc tgttcgagct ggaaaacggc cggaagagaa
     8761 tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact ggccctgccc tccaaatatg
     8821 tgaacttcct gtacctggcc agccactatg agaagctgaa gggctccccc gaggataatg
     8881 agcagaaaca gctgtttgtg gaacagcaca aacactacct ggacgagatc atcgagcaga
     8941 tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa tctggacaag gtgctgagcg
     9001 cctacaacaa gcacagagac aagcctatca gagagcaggc cgagaatatc atccacctgt
     9061 ttaccctgac caatctggga gcccctgccg ccttcaagta ctttgacacc accatcgacc
     9121 ggaagaggta caccagcacc aaagaggtgc tggacgccac cctgatccac cagagcatca
     9181 ccggcctgta cgagacacgg atcgacctgt ctcagctggg aggcgacgcc tatccctatg
     9241 acgtgcccga ttatgccagc ctgggcagcg gctcccccaa gaaaaaacgc aaggtggaag
     9301 atcctaagaa aaagcggaaa gtggacggca ttggtagtgg gagcaacgga ggtggaggct
     9361 ctggacctag gacgcgtgtg cctttgtatg gttttacttc gatttgtgga agaagacctg
     9421 agatggaagc tgctgtttcg actataccaa gattccttca atcttcctct ggttcgatgt
     9481 tagatggtcg gtttgatcct caatccgccg ctcatttctt cggtgtttac gacggccatg
     9541 gcggttctca ggtagcgaac tattgtagag agaggatgca tttggctttg gcggaggaga
     9601 tagctaagga gaaaccgatg ctctgcgatg gtgatacgtg gctggagaag tggaagaaag
     9661 ctcttttcaa ctcgttcctg agagttgact cggagattga gtcagttgcg ccggagacgg
     9721 ttgggtcaac gtcggtggtt gccgttgttt tcccgtctca catcttcgtc gctaactgcg
     9781 gtgactctag agccgttctt tgccgcggca aaactgcact tccattatcc gttgaccata
     9841 aaccggatag agaagatgaa gctgcgagga ttgaagccgc aggagggaaa gtgattcagt
     9901 ggaatggagc tcgtgttttc ggtgttctcg ccatgtcgag atccattggc gatagatact
     9961 tgaaaccatc catcattcct gatccggaag tgacggctgt gaagagagta aaagaagatg
    10021 attgtctgat tttggcgagt gacggggttt gggatgtaat gacggatgaa gaagcgtgtg
    10081 agatggcaag gaagcggatt ctcttgtggc acaagaaaaa cgcggtggct ggggatgcat
    10141 cgttgctcgc ggatgagcgg agaaaggaag ggaaagatcc tgcggcgatg tccgcggctg
    10201 agtatttgtc aaagctggcg atacagagag gaagcaaaga caacataagt gtggtggtgg
    10261 ttgatttgaa gtgactcgag ctagttaaaa gttttgttac tttatagaag aaattttgag
    10321 tttttgtttt tttttaataa ataaataaac ataaataaat tgtttgttga atttattatt
    10381 agtatgtaag tgtaaatata ataaaactta atatctattc aaattaataa ataaacctcg
    10441 atatacagac cgataaaaca catgcgtcaa ttttacgcat gattatcttt aacgtacgtc
    10501 acaatatgat tatctttcta gggttaatct agtatacgcg tatgcggccg cttaattaat
    10561 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc
    10621 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
    10681 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
    10741 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
    10801 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
    10861 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
    10921 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
    10981 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
    11041 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
    11101 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
    11161 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
    11221 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
    11281 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
    11341 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
    11401 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
    11461 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
    11521 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
    11581 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
    11641 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
    11701 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
    11761 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
    11821 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
    11881 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
    11941 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
    12001 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
    12061 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
    12121 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
    12181 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
    12241 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
    12301 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
    12361 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
    12421 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
    12481 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
    12541 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
    12601 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
    12661 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
    12721 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa
    12781 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
    12841 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
    12901 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
    12961 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
    13021 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
    13081 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
    13141 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
    13201 tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg
    13261 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
    13321 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
    13381 gcgcgcgtaa tacgactcac tatagggcga attggggcgc gccattctag attaacccta
    13441 gaaagatagt ctgcgtaaaa ttgacgcatg cattcttgaa atattgctct ctctttctaa
    13501 atagcgcgaa tccgtcgctg tgcatttagg acatctcagt cgccgcttgg agctcccgtg
    13561 aggcgtgctt gtcaatgcgg taagtgtcac tgattttgaa ctataacgac cgcgtgagtc
    13621 aaaatgacgc atgattatct tttacgtgac ttttaagatt taactcatac gataattata
    13681 ttgttatttc atgttctact tacgtgataa cttattatat atatattttc ttgttataga
    13741 tatcaactag aatgctagca tgggcccaga tctccatgg
//
