LOCUS       sequence-373708-       14233 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressed ABA-inducible dimerizing KRAB-dCas9 system, with 
            KRAB-IRES-Blasticidin resistance under CAG promoter and 
            tagBFP-dCas9-FKBP12 (F36V mutant) degron under PGK promoter in a 
            piggyBac plasmid..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14233)
  AUTHORS   Noviello G, Gjaltema RAF, Schulz EG
  TITLE     CasTuner is a degron and CRISPR/Cas-based toolkit for analog tuning 
            of endogenous gene expression.
  JOURNAL   Nat Commun. 2023 Jun 3;14(1):3225. doi: 10.1038/s41467-023-38909-4.
  PUBMED    37270532
REFERENCE   2  (bases 1 to 14233)
  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 
            Commun."; date: "2023-06-3"; volume: "14(1):3225. doi"; pages: " 
            10.1038/s41467-023-38909-4"
COMMENT     Sequence Label: pSLQ2818_Blasti_pPB:
            CAG-PYL1-KRAB-IRES-Blasti...
FEATURES             Location/Qualifiers
     source          1..14233
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   182..244
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(295..315)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(295..313)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(334..350)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(334..350)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(347..369)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    358..374
                     /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(382..388,389..406,407..412))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    427..448
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(565..582)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(736..1324)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(816..835)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1495..2286,2287..2355))
                     /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     2118..2137
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2356..2460)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2486..2941)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2623..2644)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2835..2854
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3068..3090
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3082..3099
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3083..3099
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3109..3128
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3109..3127
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   3157..3191
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        3501..3778
                     /label=chicken beta-actin promoter
     intron          3779..4795
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     4719..4741
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     4803..4822
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      4850..4859
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5477..5662
                     /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(5680..6253,6254..6256,6257..6265)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5847..5864)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6074..6093
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6281..6679
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    6717..7305
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6770..6790)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7188..7199)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7344..7363)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7398..7417
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        7459..7958
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7494..7517)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     7891..7910
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             8918..9616
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     regulatory      9632..9641
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(9638..9664,9665..9667,9668..12154,12155..12157,
                     12158..13741)
                     /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             9638..13741
                     /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             13745..13771
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             13751..13777
                     /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             13790..13810
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             13817..13837
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             13910..14233
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=FKBP-V
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
ORIGIN
        1 gctagttaaa agttttgtta ctttatagaa gaaattttga gtttttgttt ttttttaata
       61 aataaataaa cataaataaa ttgtttgttg aatttattat tagtatgtaa gtgtaaatat
      121 aataaaactt aatatctatt caaattaata aataaacctc gatatacaga ccgataaaac
      181 acatgcgtca attttacgca tgattatctt taacgtacgt cacaatatga ttatctttct
      241 agggttaatc tagtatacgc gtatgcggcc gcttaattaa tccagctttt gttcccttta
      301 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
      361 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
      421 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
      481 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
      541 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
      601 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
      661 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      721 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
      781 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      841 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
      901 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
      961 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     1021 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     1081 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     1141 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     1201 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     1261 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     1321 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     1381 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     1441 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     1501 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     1561 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     1621 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     1681 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     1741 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     1801 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     1861 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     1921 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     1981 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     2041 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     2101 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     2161 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     2221 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     2281 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     2341 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     2401 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     2461 cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg
     2521 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     2581 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     2641 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     2701 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     2761 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     2821 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     2881 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     2941 tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     3001 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     3061 gggttttccc agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta atacgactca
     3121 ctatagggcg aattggggcg cgccattcta gattaaccct agaaagatag tctgcgtaaa
     3181 attgacgcat gcattcttga aatattgctc tctctttcta aatagcgcga atccgtcgct
     3241 gtgcatttag gacatctcag tcgccgcttg gagctcccgt gaggcgtgct tgtcaatgcg
     3301 gtaagtgtca ctgattttga actataacga ccgcgtgagt caaaatgacg catgattatc
     3361 ttttacgtga cttttaagat ttaactcata cgataattat attgttattt catgttctac
     3421 ttacgtgata acttattata tatatatttt cttgttatag atatcaacta gaatgctagc
     3481 atgggcccag atctccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc
     3541 ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat
     3601 gggggcgggg gggggggggg ggcgcgcgcc rgggggggsg gggsggggsg gggggsgggg
     3661 sggggsgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc
     3721 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg
     3781 agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc
     3841 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
     3901 gggctgtaat tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag
     3961 ccttgagggg ctccgggagg gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt
     4021 gtgtgtgtgc gtggggagcg ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg
     4081 cgggcgcggc gcggggcttt gtgcgctccg cagtgtgcgc gaggggagcg cggccggggg
     4141 cggtgccccg cggtgcgggg ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg
     4201 cgtggggggg tgagcagggg gtgtgggcgc gtcggtcggg ctgcaacccc ccctgcaccc
     4261 ccctccccga gttgctgagc acggcccggc ttcgggtgcg gggctccgta cggggcgtgg
     4321 cgcggggctc gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg
     4381 gccgcctcgg gccggggrgg gctcgggggr ggggcgcggc ggcccccgga gcgccggcgg
     4441 ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg taatcgtgcg agagggcgca
     4501 gggacttcct ttgtcccaaa tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc
     4561 ctctagcggg cgcggggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg
     4621 ccttcgtgcg tcgccgcgcc gccgtcccct tctccctctc cagcctcggg gctgtccgcg
     4681 gggggacggc tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac
     4741 cggcggctct agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct
     4801 gggcaacgtg ctggttattg tgctgtctca tcattttggc aaagaattcg ccaccatggg
     4861 tgggggcgcg ccaactcaag acgaattcac ccaactctcc caatcaatcg ccgagttcca
     4921 cacgtaccaa ctcggtaacg gccgttgctc atctctccta gctcagcgaa tccacgcgcc
     4981 gccggaaaca gtatggtccg tggtgagacg tttcgatagg ccacagattt acaaacactt
     5041 catcaaaagc tgtaacgtga gtgaagattt cgagatgcga gtgggatgca cgcgcgacgt
     5101 gaacgtgata agtggattac cggcgaatac gtctcgagag agattagatc tgttggacga
     5161 tgatcggaga gtgactgggt ttagtataac cggtggtgaa cataggctga ggaattataa
     5221 atcggttacg acggttcata gatttgagaa agaagaagaa gaagaaagga tctggaccgt
     5281 tgttttggaa tcttatgttg ttgatgtacc ggaaggtaat tcggaggaag atacgagatt
     5341 gtttgctgat acggttatta gattgaatct tcagaaactt gcttcgatca ctgaagctat
     5401 gaacgaattc tcaagcgctg caggtactag tggcggtggc ggagggatgg atgctaagtc
     5461 actaactgcc tggtcccgga cactggtgac cttcaaggat gtatttgtgg acttcaccag
     5521 ggaggagtgg aagctgctgg acactgctca gcagatcgtg tacagaaatg tgatgctgga
     5581 gaactataag aacctggttt ccttgggtta tcagcttact aagccagatg tgatcctccg
     5641 gttggagaag ggagaagagc cctagattta aataattcgc ccctctccct cccccccccc
     5701 taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt
     5761 ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt
     5821 gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt
     5881 cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct
     5941 ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt
     6001 ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt
     6061 ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa
     6121 ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta cgtgtgttta
     6181 gtcgaggtta aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa
     6241 acacgatgat aatatggcca caaccggatc cggagctacc atggccaagc ctttgtctca
     6301 agaagaatcc accctcattg aaagagcaac ggctacaatc aacagcatcc ccatctctga
     6361 agactacagc gtcgccagcg cagctctctc tagcgacggc cgcatcttca ctggtgtcaa
     6421 tgtatatcat tttactgggg gaccttgtgc agaactcgtg gtgctgggca ctgctgctgc
     6481 tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga aatgagaaca ggggcatctt
     6541 gagcccctgc ggacggtgcc gacaggtgct tctcgatctg catcctggga tcaaagccat
     6601 agtgaaggac agtgatggac agccgacggc agttgggatt cgtgaattgc tgccctctgg
     6661 ttatgtgtgg gagggctaag tttaaaccat gcaagcttga tatcaagctt atcgataatc
     6721 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     6781 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     6841 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     6901 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     6961 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     7021 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     7081 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     7141 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     7201 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     7261 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcct tgtttattgc
     7321 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     7381 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat
     7441 cctgcaggta attctaccgg gtaggggagg cgcttttccc aaggcagtct ggagcatgcg
     7501 ctttagcagc cccgctgggc acttggcgct acacaagtgg cctctggcct cgcacacatt
     7561 ccacatccac cggtaggcgc caaccggctc cgttctttgg tggccccttc gcgccacctt
     7621 ctactcctcc cctagtcagg aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt
     7681 gacaaatgga agtagcacgt ctcactaggc tcgtgcagat ggacagcacc gctgagcaat
     7741 ggaagcgggt aggcctttgg ggcagcggcc aatagcagct ttgctccttc gctttctggg
     7801 ctcagaggct gggaaggggt gggtccgggg gcgggctcag gggcgggctc aggggcgggg
     7861 cgggcgcccg aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg
     7921 ccgcgctgtt ctcctcttcc tcatctccgg gcctttcgac ctgcagccca agcttgtcga
     7981 cgccaccatg acgcgtgtgc ctttgtatgg ttttacttcg atttgtggaa gaagacctga
     8041 gatggaagct gctgtttcga ctataccaag attccttcaa tcttcctctg gttcgatgtt
     8101 agatggtcgg tttgatcctc aatccgccgc tcatttcttc ggtgtttacg acggccatgg
     8161 cggttctcag gtagcgaact attgtagaga gaggatgcat ttggctttgg cggaggagat
     8221 agctaaggag aaaccgatgc tctgcgatgg tgatacgtgg ctggagaagt ggaagaaagc
     8281 tcttttcaac tcgttcctga gagttgactc ggagattgag tcagttgcgc cggagacggt
     8341 tgggtcaacg tcggtggttg ccgttgtttt cccgtctcac atcttcgtcg ctaactgcgg
     8401 tgactctaga gccgttcttt gccgcggcaa aactgcactt ccattatccg ttgaccataa
     8461 accggataga gaagatgaag ctgcgaggat tgaagccgca ggagggaaag tgattcagtg
     8521 gaatggagct cgtgttttcg gtgttctcgc catgtcgaga tccattggcg atagatactt
     8581 gaaaccatcc atcattcctg atccggaagt gacggctgtg aagagagtaa aagaagatga
     8641 ttgtctgatt ttggcgagtg acggggtttg ggatgtaatg acggatgaag aagcgtgtga
     8701 gatggcaagg aagcggattc tcttgtggca caagaaaaac gcggtggctg gggatgcatc
     8761 gttgctcgcg gatgagcgga gaaaggaagg gaaagatcct gcggcgatgt ccgcggctga
     8821 gtatttgtca aagctggcga tacagagagg aagcaaagac aacataagtg tggtggtggt
     8881 tgatttgaag catatgtcaa gcgctgcagg tgctaccatg agcgagctga ttaaggagaa
     8941 catgcacatg aagctgtaca tggagggcac cgtggacaac catcacttca agtgcacatc
     9001 cgagggcgaa ggcaagccct acgagggcac ccagaccatg agaatcaagg tggtcgaggg
     9061 cggccctctc cccttcgcct tcgacatcct ggctactagc ttcctctacg gcagcaagac
     9121 cttcatcaac cacacccagg gcatccccga cttcttcaag cagtccttcc ctgagggctt
     9181 cacatgggag agagtcacca catacgaaga cgggggcgtg ctgaccgcta cccaggacac
     9241 cagcctccag gacggctgcc tcatctacaa cgtcaagatc agaggggtga acttcacatc
     9301 caacggccct gtgatgcaga agaaaacact cggctgggag gccttcaccg agacgctgta
     9361 ccccgctgac ggcggcctgg aaggcagaaa cgacatggcc ctgaagctcg tgggcgggag
     9421 ccatctgatc gcaaacatca agaccacata tagatccaag aaacccgcta agaacctcaa
     9481 gatgcctggc gtctactatg tggactacag actggaaaga atcaaggagg ccaacaacga
     9541 gacctacgtc gagcagcacg aggtggcagt ggccagatac tgcgacctcc ctagcaaact
     9601 ggggcacaag cttaattcaa gcgctgcagg tgccaccatg gacaagaagt acagcatcgg
     9661 cctggccatc ggcaccaact ctgtgggctg ggccgtgatc accgacgagt acaaggtgcc
     9721 cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga agaacctgat
     9781 cggcgccctg ctgttcgaca gcggagaaac agccgaggcc acccggctga agagaaccgc
     9841 cagaagaaga tacaccagac ggaagaaccg gatctgctat ctgcaagaga tcttcagcaa
     9901 cgagatggcc aaggtggacg acagcttctt ccacagactg gaagagtcct tcctggtgga
     9961 agaggataag aagcacgagc ggcaccccat cttcggcaac atcgtggacg aggtggccta
    10021 ccacgagaag taccccacca tctaccacct gagaaagaaa ctggtggaca gcaccgacaa
    10081 ggccgacctg cggctgatct atctggccct ggcccacatg atcaagttcc ggggccactt
    10141 cctgatcgag ggcgacctga accccgacaa cagcgacgtg gacaagctgt tcatccagct
    10201 ggtgcagacc tacaaccagc tgttcgagga aaaccccatc aacgccagcg gcgtggacgc
    10261 caaggccatc ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc tgatcgccca
    10321 gctgcccggc gagaagaaga atggcctgtt cggcaacctg attgccctga gcctgggcct
    10381 gacccccaac ttcaagagca acttcgacct ggccgaggat gccaaactgc agctgagcaa
    10441 ggacacctac gacgacgacc tggacaacct gctggcccag atcggcgacc agtacgccga
    10501 cctgtttctg gccgccaaga acctgtccga cgccatcctg ctgagcgaca tcctgagagt
    10561 gaacaccgag atcaccaagg cccccctgag cgcctctatg atcaagagat acgacgagca
    10621 ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg agaagtacaa
    10681 agagattttc ttcgaccaga gcaagaacgg ctacgccggc tacatcgatg gcggagccag
    10741 ccaggaagag ttctacaagt tcatcaagcc catcctggaa aagatggacg gcaccgagga
    10801 actgctcgtg aagctgaaca gagaggacct gctgcggaag cagcggacct tcgacaacgg
    10861 cagcatcccc caccagatcc acctgggaga gctgcacgcc attctgcggc ggcaggaaga
    10921 tttttaccca ttcctgaagg acaaccggga aaagatcgag aagatcctga ccttccgcat
    10981 cccctactac gtgggccctc tggccagggg aaacagcaga ttcgcctgga tgaccagaaa
    11041 gagcgaggaa accatcaccc cctggaactt cgaggaagtg gtggacaagg gcgccagcgc
    11101 ccagagcttc atcgagcgga tgaccaactt cgataagaac ctgcccaacg agaaggtgct
    11161 gcccaagcac agcctgctgt acgagtactt caccgtgtac aacgagctga ccaaagtgaa
    11221 atacgtgacc gagggaatga gaaagcccgc cttcctgagc ggcgagcaga aaaaagccat
    11281 cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga aagaggacta
    11341 cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag atcggttcaa
    11401 cgcctccctg ggcacatacc acgatctgct gaaaattatc aaggacaagg acttcctgga
    11461 caatgaggaa aacgaggaca ttctggaaga tatcgtgctg accctgacac tgtttgagga
    11521 cagagagatg atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg acaaagtgat
    11581 gaagcagctg aagcggcgga gatacaccgg ctggggcagg ctgagccgga agctgatcaa
    11641 cggcatccgg gacaagcagt ccggcaagac aatcctggat ttcctgaagt ccgacggctt
    11701 cgccaacaga aacttcatgc agctgatcca cgacgacagc ctgaccttta aagaggacat
    11761 ccagaaagcc caggtgtccg gccagggcga tagcctgcac gagcacattg ccaatctggc
    11821 cggcagcccc gccattaaga agggcatcct gcagacagtg aaggtggtgg acgagctcgt
    11881 gaaagtgatg ggccggcaca agcccgagaa catcgtgatc gaaatggcca gagagaacca
    11941 gaccacccag aagggacaga agaacagccg cgagagaatg aagcggatcg aagagggcat
    12001 caaagagctg ggcagccaga tcctgaaaga acaccccgtg gaaaacaccc agctgcagaa
    12061 cgagaagctg tacctgtact acctgcagaa tgggcgggat atgtacgtgg accaggaact
    12121 ggacatcaac cggctgtccg actacgatgt ggacgctatc gtgcctcaga gctttctgaa
    12181 ggacgactcc atcgataaca aagtgctgac tcggagcgac aagaaccggg gcaagagcga
    12241 caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac tactggcgcc agctgctgaa
    12301 tgccaagctg attacccaga ggaagttcga caatctgacc aaggccgaga gaggcggcct
    12361 gagcgaactg gataaggccg gcttcatcaa gagacagctg gtggaaaccc ggcagatcac
    12421 aaagcacgtg gcacagatcc tggactcccg gatgaacact aagtacgacg agaacgacaa
    12481 actgatccgg gaagtgaaag tgatcaccct gaagtccaag ctggtgtccg atttccggaa
    12541 ggatttccag ttttacaaag tgcgcgagat caacaactac caccacgccc acgacgccta
    12601 cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac cctaagctgg aaagcgagtt
    12661 cgtgtacggc gactacaagg tgtacgacgt gcggaagatg atcgccaaga gcgagcagga
    12721 aatcggcaag gctaccgcca agtacttctt ctacagcaac atcatgaact ttttcaagac
    12781 cgagattacc ctggccaacg gcgagatccg gaagcggcct ctgatcgaga caaacggcga
    12841 aacaggcgag atcgtgtggg ataagggccg ggactttgcc accgtgcgga aagtgctgtc
    12901 tatgccccaa gtgaatatcg tgaaaaagac cgaggtgcag acaggcggct tcagcaaaga
    12961 gtctatcctg cccaagagga acagcgacaa gctgatcgcc agaaagaagg actgggaccc
    13021 taagaagtac ggcggcttcg acagccccac cgtggcctat tctgtgctgg tggtggccaa
    13081 agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa gagctgctgg ggatcaccat
    13141 catggaaaga agcagcttcg agaagaatcc catcgacttt ctggaagcca agggctacaa
    13201 agaagtgaaa aaggacctga tcatcaagct gcctaagtac tccctgttcg agctggaaaa
    13261 cggccggaag agaatgctgg cctctgccgg cgaactgcag aagggaaacg aactggccct
    13321 gccctccaaa tatgtgaact tcctgtacct ggccagccac tatgagaagc tgaagggctc
    13381 ccccgaggat aatgagcaga aacagctgtt tgtggaacag cacaaacact acctggacga
    13441 gatcatcgag cagatcagcg agttctccaa gagagtgatc ctggccgacg ctaatctgga
    13501 caaggtgctg agcgcctaca acaagcacag agacaagcct atcagagagc aggccgagaa
    13561 tatcatccac ctgtttaccc tgaccaatct gggagcccct gccgccttca agtactttga
    13621 caccaccatc gaccggaaga ggtacaccag caccaaagag gtgctggacg ccaccctgat
    13681 ccaccagagc atcaccggcc tgtacgagac acggatcgac ctgtctcagc tgggaggcga
    13741 cgcctatccc tatgacgtgc ccgattatgc cagcctgggc agcggctccc ccaagaaaaa
    13801 acgcaaggtg gaagatccta agaaaaagcg gaaagtggac ggcattggta gtgggagcaa
    13861 cggaggcgga tctggaggta gtggtggatc tggaggttct ggtggctccg gagttcaggt
    13921 ggaaaccatc tccccaggag acgggcgcac cttccccaag cgcggccaga cctgcgtggt
    13981 gcactacacc gggatgcttg aagatggaaa gaaagttgat tcctcccggg acagaaacaa
    14041 gccctttaag tttatgctag gcaagcagga ggtgatccga ggctgggaag aaggggttgc
    14101 ccagatgagt gtgggtcaga gagccaaact gactatatct ccagattatg cctatggtgc
    14161 cactgggcac ccaggcatca tcccaccaca tgccactctc gtcttcgatg tggagcttct
    14221 aaaactggaa tag
//
