LOCUS       sequence-160888-       11836 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses direct fusion KRAB-Sp dCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11836)
  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 11836)
  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: pSLQ2815 pPB: CAG-Puro-WPRE PGK-KRAB-tagBFP-Sp...
FEATURES             Location/Qualifiers
     source          1..11836
                     /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"
     CDS             1370..1969
                     /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(1370..1389)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1866..1886
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    2007..2595
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2060..2080)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2478..2489)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        2616..3115
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2651..2674)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3048..3067
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      3139..3148
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3175..3360
                     /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"
     CDS             3388..4086
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     regulatory      4102..4111
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4108..4134,4135..4137,4138..6624,6625..6627,
                     6628..8211)
                     /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             4108..8211
                     /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             8215..8241
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             8221..8247
                     /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             8260..8280
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8287..8307
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     repeat_region   8524..8586
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(8637..8657)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(8637..8655)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8676..8692)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8676..8692)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8689..8711)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8700..8716
                     /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(8724..8730,8731..8748,8749..8754))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8769..8790
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8907..8924)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9078..9666)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9158..9177)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9837..10628,10629..10697))
                     /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     10460..10479
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10698..10802)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(10828..11283)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10965..10986)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     11177..11196
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11410..11432
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     11424..11441
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     11425..11441
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11451..11470
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        11451..11469
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   11499..11533
                     /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 accatgacca tgaccgagta
     1381 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc
     1441 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga
     1501 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
     1561 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
     1621 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
     1681 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
     1741 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
     1801 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
     1861 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
     1921 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgag tttaaaccat
     1981 gcaagcttga tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat
     2041 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     2101 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     2161 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     2221 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     2281 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     2341 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     2401 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     2461 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     2521 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     2581 gggccgcctc cccgcatcct gcaggtaatt ctaccgggta ggggaggcgc ttttcccaag
     2641 gcagtctgga gcatgcgctt tagcagcccc gctgggcact tggcgctaca caagtggcct
     2701 ctggcctcgc acacattcca catccaccgg taggcgccaa ccggctccgt tctttggtgg
     2761 ccccttcgcg ccaccttcta ctcctcccct agtcaggaag ttcccccccg ccccgcagct
     2821 cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc actaggctcg tgcagatgga
     2881 cagcaccgct gagcaatgga agcgggtagg cctttggggc agcggccaat agcagctttg
     2941 ctccttcgct ttctgggctc agaggctggg aaggggtggg tccgggggcg ggctcagggg
     3001 cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga ggcccggcat tctgcacgct
     3061 tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca tctccgggcc tttcgacctg
     3121 cagcccaagc ttgtcgacgc caccatggat gctaagtcac taactgcctg gtcccggaca
     3181 ctggtgacct tcaaggatgt atttgtggac ttcaccaggg aggagtggaa gctgctggac
     3241 actgctcagc agatcgtgta cagaaatgtg atgctggaga actataagaa cctggtttcc
     3301 ttgggttatc agcttactaa gccagatgtg atcctccggt tggagaaggg agaagagccc
     3361 catatgtcaa gcgctgcagg tgctaccatg agcgagctga ttaaggagaa catgcacatg
     3421 aagctgtaca tggagggcac cgtggacaac catcacttca agtgcacatc cgagggcgaa
     3481 ggcaagccct acgagggcac ccagaccatg agaatcaagg tggtcgaggg cggccctctc
     3541 cccttcgcct tcgacatcct ggctactagc ttcctctacg gcagcaagac cttcatcaac
     3601 cacacccagg gcatccccga cttcttcaag cagtccttcc ctgagggctt cacatgggag
     3661 agagtcacca catacgaaga cgggggcgtg ctgaccgcta cccaggacac cagcctccag
     3721 gacggctgcc tcatctacaa cgtcaagatc agaggggtga acttcacatc caacggccct
     3781 gtgatgcaga agaaaacact cggctgggag gccttcaccg agacgctgta ccccgctgac
     3841 ggcggcctgg aaggcagaaa cgacatggcc ctgaagctcg tgggcgggag ccatctgatc
     3901 gcaaacatca agaccacata tagatccaag aaacccgcta agaacctcaa gatgcctggc
     3961 gtctactatg tggactacag actggaaaga atcaaggagg ccaacaacga gacctacgtc
     4021 gagcagcacg aggtggcagt ggccagatac tgcgacctcc ctagcaaact ggggcacaag
     4081 cttaattcaa gcgctgcagg tgccaccatg gacaagaagt acagcatcgg cctggccatc
     4141 ggcaccaact ctgtgggctg ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa
     4201 ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga agaacctgat cggcgccctg
     4261 ctgttcgaca gcggagaaac agccgaggcc acccggctga agagaaccgc cagaagaaga
     4321 tacaccagac ggaagaaccg gatctgctat ctgcaagaga tcttcagcaa cgagatggcc
     4381 aaggtggacg acagcttctt ccacagactg gaagagtcct tcctggtgga agaggataag
     4441 aagcacgagc ggcaccccat cttcggcaac atcgtggacg aggtggccta ccacgagaag
     4501 taccccacca tctaccacct gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg
     4561 cggctgatct atctggccct ggcccacatg atcaagttcc ggggccactt cctgatcgag
     4621 ggcgacctga accccgacaa cagcgacgtg gacaagctgt tcatccagct ggtgcagacc
     4681 tacaaccagc tgttcgagga aaaccccatc aacgccagcg gcgtggacgc caaggccatc
     4741 ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc tgatcgccca gctgcccggc
     4801 gagaagaaga atggcctgtt cggcaacctg attgccctga gcctgggcct gacccccaac
     4861 ttcaagagca acttcgacct ggccgaggat gccaaactgc agctgagcaa ggacacctac
     4921 gacgacgacc tggacaacct gctggcccag atcggcgacc agtacgccga cctgtttctg
     4981 gccgccaaga acctgtccga cgccatcctg ctgagcgaca tcctgagagt gaacaccgag
     5041 atcaccaagg cccccctgag cgcctctatg atcaagagat acgacgagca ccaccaggac
     5101 ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg agaagtacaa agagattttc
     5161 ttcgaccaga gcaagaacgg ctacgccggc tacatcgatg gcggagccag ccaggaagag
     5221 ttctacaagt tcatcaagcc catcctggaa aagatggacg gcaccgagga actgctcgtg
     5281 aagctgaaca gagaggacct gctgcggaag cagcggacct tcgacaacgg cagcatcccc
     5341 caccagatcc acctgggaga gctgcacgcc attctgcggc ggcaggaaga tttttaccca
     5401 ttcctgaagg acaaccggga aaagatcgag aagatcctga ccttccgcat cccctactac
     5461 gtgggccctc tggccagggg aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa
     5521 accatcaccc cctggaactt cgaggaagtg gtggacaagg gcgccagcgc ccagagcttc
     5581 atcgagcgga tgaccaactt cgataagaac ctgcccaacg agaaggtgct gcccaagcac
     5641 agcctgctgt acgagtactt caccgtgtac aacgagctga ccaaagtgaa atacgtgacc
     5701 gagggaatga gaaagcccgc cttcctgagc ggcgagcaga aaaaagccat cgtggacctg
     5761 ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga aagaggacta cttcaagaaa
     5821 atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg
     5881 ggcacatacc acgatctgct gaaaattatc aaggacaagg acttcctgga caatgaggaa
     5941 aacgaggaca ttctggaaga tatcgtgctg accctgacac tgtttgagga cagagagatg
     6001 atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg acaaagtgat gaagcagctg
     6061 aagcggcgga gatacaccgg ctggggcagg ctgagccgga agctgatcaa cggcatccgg
     6121 gacaagcagt ccggcaagac aatcctggat ttcctgaagt ccgacggctt cgccaacaga
     6181 aacttcatgc agctgatcca cgacgacagc ctgaccttta aagaggacat ccagaaagcc
     6241 caggtgtccg gccagggcga tagcctgcac gagcacattg ccaatctggc cggcagcccc
     6301 gccattaaga agggcatcct gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg
     6361 ggccggcaca agcccgagaa catcgtgatc gaaatggcca gagagaacca gaccacccag
     6421 aagggacaga agaacagccg cgagagaatg aagcggatcg aagagggcat caaagagctg
     6481 ggcagccaga tcctgaaaga acaccccgtg gaaaacaccc agctgcagaa cgagaagctg
     6541 tacctgtact acctgcagaa tgggcgggat atgtacgtgg accaggaact ggacatcaac
     6601 cggctgtccg actacgatgt ggacgctatc gtgcctcaga gctttctgaa ggacgactcc
     6661 atcgataaca aagtgctgac tcggagcgac aagaaccggg gcaagagcga caacgtgccc
     6721 tccgaagagg tcgtgaagaa gatgaagaac tactggcgcc agctgctgaa tgccaagctg
     6781 attacccaga ggaagttcga caatctgacc aaggccgaga gaggcggcct gagcgaactg
     6841 gataaggccg gcttcatcaa gagacagctg gtggaaaccc ggcagatcac aaagcacgtg
     6901 gcacagatcc tggactcccg gatgaacact aagtacgacg agaacgacaa actgatccgg
     6961 gaagtgaaag tgatcaccct gaagtccaag ctggtgtccg atttccggaa ggatttccag
     7021 ttttacaaag tgcgcgagat caacaactac caccacgccc acgacgccta cctgaacgcc
     7081 gtcgtgggaa ccgccctgat caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc
     7141 gactacaagg tgtacgacgt gcggaagatg atcgccaaga gcgagcagga aatcggcaag
     7201 gctaccgcca agtacttctt ctacagcaac atcatgaact ttttcaagac cgagattacc
     7261 ctggccaacg gcgagatccg gaagcggcct ctgatcgaga caaacggcga aacaggcgag
     7321 atcgtgtggg ataagggccg ggactttgcc accgtgcgga aagtgctgtc tatgccccaa
     7381 gtgaatatcg tgaaaaagac cgaggtgcag acaggcggct tcagcaaaga gtctatcctg
     7441 cccaagagga acagcgacaa gctgatcgcc agaaagaagg actgggaccc taagaagtac
     7501 ggcggcttcg acagccccac cgtggcctat tctgtgctgg tggtggccaa agtggaaaag
     7561 ggcaagtcca agaaactgaa gagtgtgaaa gagctgctgg ggatcaccat catggaaaga
     7621 agcagcttcg agaagaatcc catcgacttt ctggaagcca agggctacaa agaagtgaaa
     7681 aaggacctga tcatcaagct gcctaagtac tccctgttcg agctggaaaa cggccggaag
     7741 agaatgctgg cctctgccgg cgaactgcag aagggaaacg aactggccct gccctccaaa
     7801 tatgtgaact tcctgtacct ggccagccac tatgagaagc tgaagggctc ccccgaggat
     7861 aatgagcaga aacagctgtt tgtggaacag cacaaacact acctggacga gatcatcgag
     7921 cagatcagcg agttctccaa gagagtgatc ctggccgacg ctaatctgga caaggtgctg
     7981 agcgcctaca acaagcacag agacaagcct atcagagagc aggccgagaa tatcatccac
     8041 ctgtttaccc tgaccaatct gggagcccct gccgccttca agtactttga caccaccatc
     8101 gaccggaaga ggtacaccag caccaaagag gtgctggacg ccaccctgat ccaccagagc
     8161 atcaccggcc tgtacgagac acggatcgac ctgtctcagc tgggaggcga cgcctatccc
     8221 tatgacgtgc ccgattatgc cagcctgggc agcggctccc ccaagaaaaa acgcaaggtg
     8281 gaagatccta agaaaaagcg gaaagtggac ggcattggta gtgggagcaa cggatgactc
     8341 gagctagtta aaagttttgt tactttatag aagaaatttt gagtttttgt ttttttttaa
     8401 taaataaata aacataaata aattgtttgt tgaatttatt attagtatgt aagtgtaaat
     8461 ataataaaac ttaatatcta ttcaaattaa taaataaacc tcgatataca gaccgataaa
     8521 acacatgcgt caattttacg catgattatc tttaacgtac gtcacaatat gattatcttt
     8581 ctagggttaa tctagtatac gcgtatgcgg ccgcttaatt aatccagctt ttgttccctt
     8641 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     8701 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     8761 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     8821 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     8881 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     8941 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     9001 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     9061 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     9121 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     9181 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     9241 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     9301 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     9361 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     9421 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     9481 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     9541 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     9601 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     9661 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     9721 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     9781 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     9841 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     9901 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     9961 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
    10021 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
    10081 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
    10141 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
    10201 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
    10261 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
    10321 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
    10381 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
    10441 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
    10501 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
    10561 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
    10621 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
    10681 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
    10741 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
    10801 cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg
    10861 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
    10921 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
    10981 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
    11041 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
    11101 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
    11161 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
    11221 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
    11281 cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
    11341 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
    11401 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg taatacgact
    11461 cactataggg cgaattgggg cgcgccattc tagattaacc ctagaaagat agtctgcgta
    11521 aaattgacgc atgcattctt gaaatattgc tctctctttc taaatagcgc gaatccgtcg
    11581 ctgtgcattt aggacatctc agtcgccgct tggagctccc gtgaggcgtg cttgtcaatg
    11641 cggtaagtgt cactgatttt gaactataac gaccgcgtga gtcaaaatga cgcatgatta
    11701 tcttttacgt gacttttaag atttaactca tacgataatt atattgttat ttcatgttct
    11761 acttacgtga taacttatta tatatatatt ttcttgttat agatatcaac tagaatgcta
    11821 gcatgggccc agatct
//
