LOCUS       sequence-237262-       13465 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  piggyBac transposon Destination vector for dox-inducible expression 
            of Gateway cloned elements (inducible mCherry). KRAB-dCas9 is 
            constitutively expressed..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13465)
  AUTHORS   Kagawa H, Shimamoto R, Kim SI, Oceguera-Yanez F, Yamamoto T, 
            Schroeder T, Woltjen K
  TITLE     OVOL1 Influences the Determination and Expansion of iPSC 
            Reprogramming Intermediates.
  JOURNAL   Stem Cell Reports. 2018 Dec 28. pii: S2213-6711(18)30527-7. doi: 
            10.1016/j.stemcr.2018.12.008.
  PUBMED    30639212
REFERENCE   2  (bases 1 to 13465)
  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: "Stem Cell 
            Reports. 2018 Dec 28. pii: S2213-6711(18)30527-7. doi: 
            10.1016/j.stemcr.2018.12.008."
COMMENT     Sequence Label: PB-TAC-KRAB-dCas9
FEATURES             Location/Qualifiers
     source          1..13465
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(28..456)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(138..159)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     350..369
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     583..605
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     597..614
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     598..614
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     621..640
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        621..639
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   706..768
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    1140..1158
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1177..1195
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1214..1232
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1251..1269
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1288..1306
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1325..1343
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1362..1380
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1399..1417
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1439..1463
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    1543..1667
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(1692..1697,1698..1715,1716..1722)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1776..2435
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1842..1861)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2777..3082
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     2961..2980
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3123..3247)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    join(3289..3863,3864..3866,3867..3875)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3456..3473)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3683..3702
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3876..4586
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4021..4039)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4289..4308
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(4598..4617)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4663..4718
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4717..4736)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     enhancer        5155..5440
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        5442..5719
                     /label=chicken beta-actin promoter
     intron          5720..5947
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      5959..5968
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5995..6180
                     /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             join(6202..6228,6229..6231,6232..8718,8719..8721,
                     8722..10305)
                     /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             6202..10305
                     /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             10309..10335
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             10315..10341
                     /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             10354..10374
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10381..10401
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(10468..10485)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10474..10681
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   11092..11126
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(11275..11293)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(11314..11330)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11314..11330)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11327..11349)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11338..11354
                     /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(11362..11368,11369..11386,11387..11392))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11407..11428
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11545..11562)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11716..12304)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11796..11815)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12475..13266,13267..13335))
                     /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     13098..13117
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13336..13440)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
       61 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
      121 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
      181 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
      241 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
      301 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
      361 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
      421 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca
      481 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
      541 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
      601 aaacgacggc cagtgaattg taatacgact cactataggg cgaattggag ctcggtattc
      661 acgacagcag gctgaataat aaaaaaatta gaaactatta tttaacccta gaaagataat
      721 catattgtga cgtacgttaa agataatcat gcgtaaaatt gacgcatgtg ttttatcggt
      781 ctgtatatcg aggtttattt attaatttga atagatatta agttttatta tatttacact
      841 tacatactaa taataaattc aacaaacaat ttatttatgt ttatttattt attaaaaaaa
      901 aacaaaaact caaaatttct tctataaagt aacaaaactt ttaaacattc tctcttttac
      961 aaaaataaac ttattttgta ctttaaaaac agtcatgttg tattataaaa taagtaatta
     1021 gcttaaccta tacataatag aaacaaatta tacttattag tcagtcagaa acaactttgg
     1081 cacatatcaa tattatgctc tcgttaatta actcgaggga tccaagctgg ccgcacgtct
     1141 ccctatcagt gatagagaag tcgacacgtc tcgagctccc tatcagtgat agagaaggta
     1201 cgtctagaac gtctccctat cagtgataga gaagtcgaca cgtctcgagc tccctatcag
     1261 tgatagagaa ggtacgtcta gaacgtctcc ctatcagtga tagagaagtc gacacgtctc
     1321 gagctcccta tcagtgatag agaaggtacg tctagaacgt ctccctatca gtgatagaga
     1381 agtcgacacg tctcgagctc cctatcagtg atagagaagg taccccctat ataagcagag
     1441 ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata
     1501 gaagacaccg ggaccgatcc agcctccgcg gtggcggcca tcacaagttt gtacaaaaaa
     1561 gctgaacgag aaacgtaaaa tgatataaat atcaatatat taaattagat tttgcataaa
     1621 aaacagacta cataatactg taaaacacaa catatccagt cactatggcg gccgcattag
     1681 gcaccccagg ctttacactt tatgcttccg gctcgtataa tgtgtggatt ttgagttagg
     1741 atccgtcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc actggatata
     1801 ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt cagtcagttg
     1861 ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta aagaccgtaa
     1921 agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc ctgatgaatg
     1981 ctcatccgga attccgtatg gcaatgaaag acggtgagct ggtgatatgg gatagtgttc
     2041 acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat
     2101 accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg tgttacggtg
     2161 aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc tcagccaatc
     2221 cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac ttcttcgccc
     2281 ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg ccgctggcga
     2341 ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt aatgaattac
     2401 aacagtactg cgatgagtgg cagggcgggg cgtaaacgcg tggatccggc ttactaaaag
     2461 ccagataaca gtatgcgtat ttgcgcgctg atttttgcgg tataagaata tatactgata
     2521 tgtatacccg aagtatgtca aaaagaggta tgctatgaag cagcgtatta cagtgacagt
     2581 tgacagcgac agctatcagt tgctcaaggc atatatgatg tcaatatctc cggtctggta
     2641 agcacaacca tgcagaatga agcccgtcgt ctgcgtgccg aacgctggaa agcggaaaat
     2701 caggaaggga tggctgaggt cgcccggttt attgaaatga acggctcttt tgctgacgag
     2761 aacaggggct ggtgaaatgc agtttaaggt ttacacctat aaaagagaga gccgttatcg
     2821 tctgtttgtg gatgtacaga gtgatattat tgacacgccc gggcgacgga tggtgatccc
     2881 cctggccagt gcacgtctgc tgtcagataa agtctcccgt gaactttacc cggtggtgca
     2941 tatcggggat gaaagctggc gcatgatgac caccgatatg gccagtgtgc cggtctccgt
     3001 tatcggggaa gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa acgccattaa
     3061 cctgatgttc tggggaatat aaatgtcagg ctcccttata cacagccagt ctgcaggtcg
     3121 accatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt ttatgcaaaa
     3181 tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcagct ttcttgtaca
     3241 aagtggtgat ctagctaaaa cgcggcctcg aatcgaattc cgcccccccc cctctccctc
     3301 ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta
     3361 tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc
     3421 tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct
     3481 gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt
     3541 agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa
     3601 gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg
     3661 gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga
     3721 tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac
     3781 atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt
     3841 cctttgaaaa acacgatgat aatatggcca caaccatggt gagcaagggc gaggaggata
     3901 acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg
     3961 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
     4021 ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc
     4081 agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga
     4141 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
     4201 tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc
     4261 tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg
     4321 aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga
     4381 ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca
     4441 agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc
     4501 acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg
     4561 gcggcatgga cgagctgtac aagtaaagcg gccgcgatct ttttccctct gccaaaaatt
     4621 atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg aaatttattt
     4681 tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg
     4741 caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca tatgcccata
     4801 tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg aaacagcccc
     4861 ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt ttttttatat
     4921 tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat gttttactag
     4981 ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt ctcttatgga
     5041 gatccctcga cctgcaggaa ttcgattact taagcctagg atccagatct gtacattcga
     5101 atcgatgaat tcgttaacca tggaagcttc tgcagcggcc gcactagagg tacccgttac
     5161 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     5221 aatagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     5281 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     5341 tgacggtaaa tggcccgcct ggcattgtgc ccagtacatg accttatggg actttcctac
     5401 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtcgaggtga gccccacgtt
     5461 ctgcttcact ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt
     5521 ttaattattt tgtgcagcga tgggggcggg gggggggggg gggcgcgcgc crggsggggs
     5581 ggggsggggs grggggsggg gsggggcgag gcggagaggt gcggcggcag ccaatcagag
     5641 cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg cggcggcggc cctataaaaa
     5701 gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc
     5761 cgccgcctcg cgccgcccgc cccggctctg actgaccgcg ttactcccac aggtgagcgg
     5821 gcgggacggc ccttctcctc cgggctgtaa ttagctgagc aagaggtaag ggtttaaggg
     5881 atggttggtt ggtggggtat taatgtttaa ttacctggag cacctgcctg aaatcacttt
     5941 ttttcaggtt ggaccggtgc caccatggat gctaagtcac taactgcctg gtcccggaca
     6001 ctggtgacct tcaaggatgt atttgtggac ttcaccaggg aggagtggaa gctgctggac
     6061 actgctcagc agatcgtgta cagaaatgtg atgctggaga actataagaa cctggtttcc
     6121 ttgggttatc agcttactaa gccagatgtg atcctccggt tggagaaggg agaagagccc
     6181 ggcggttccg gcggagggtc gatggacaag aagtacagca tcggcctggc catcggcacc
     6241 aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag
     6301 gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggcgc cctgctgttc
     6361 gacagcggag aaacagccga ggccacccgg ctgaagagaa ccgccagaag aagatacacc
     6421 agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg
     6481 gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac
     6541 gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc
     6601 accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg
     6661 atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac
     6721 ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac
     6781 cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc catcctgtct
     6841 gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc cggcgagaag
     6901 aagaatggcc tgttcggcaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag
     6961 agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac ctacgacgac
     7021 gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc
     7081 aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc
     7141 aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca ggacctgacc
     7201 ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac
     7261 cagagcaaga acggctacgc cggctacatc gatggcggag ccagccagga agagttctac
     7321 aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg
     7381 aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat cccccaccag
     7441 atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg
     7501 aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc
     7561 cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc
     7621 accccctgga acttcgagga agtggtggac aagggcgcca gcgcccagag cttcatcgag
     7681 cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg
     7741 ctgtacgagt acttcaccgt gtacaacgag ctgaccaaag tgaaatacgt gaccgaggga
     7801 atgagaaagc ccgccttcct gagcggcgag cagaaaaaag ccatcgtgga cctgctgttc
     7861 aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag
     7921 tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca
     7981 taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag
     8041 gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag
     8101 gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg
     8161 cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat ccgggacaag
     8221 cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc
     8281 atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggtg
     8341 tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt
     8401 aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg
     8461 cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac ccagaaggga
     8521 cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc
     8581 cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa gctgtacctg
     8641 tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat caaccggctg
     8701 tccgactacg atgtggacgc tatcgtgcct cagagctttc tgaaggacga ctccatcgat
     8761 aacaaagtgc tgactcggag cgacaagaac cggggcaaga gcgacaacgt gccctccgaa
     8821 gaggtcgtga agaagatgaa gaactactgg cgccagctgc tgaatgccaa gctgattacc
     8881 cagaggaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga actggataag
     8941 gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca cgtggcacag
     9001 atcctggact cccggatgaa cactaagtac gacgagaacg acaaactgat ccgggaagtg
     9061 aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt ccagttttac
     9121 aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg
     9181 ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta cggcgactac
     9241 aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg caaggctacc
     9301 gccaagtact tcttctacag caacatcatg aactttttca agaccgagat taccctggcc
     9361 aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaacagg cgagatcgtg
     9421 tgggataagg gccgggactt tgccaccgtg cggaaagtgc tgtctatgcc ccaagtgaat
     9481 atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat cctgcccaag
     9541 aggaacagcg acaagctgat cgccagaaag aaggactggg accctaagaa gtacggcggc
     9601 ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag
     9661 tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga aagaagcagc
     9721 ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt gaaaaaggac
     9781 ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg gaagagaatg
     9841 ctggcctctg ccggcgaact gcagaaggga aacgaactgg ccctgccctc caaatatgtg
     9901 aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga ggataatgag
     9961 cagaaacagc tgtttgtgga acagcacaaa cactacctgg acgagatcat cgagcagatc
    10021 agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaggt gctgagcgcc
    10081 tacaacaagc acagagacaa gcctatcaga gagcaggccg agaatatcat ccacctgttt
    10141 accctgacca atctgggagc ccctgccgcc ttcaagtact ttgacaccac catcgaccgg
    10201 aagaggtaca ccagcaccaa agaggtgctg gacgccaccc tgatccacca gagcatcacc
    10261 ggcctgtacg agacacggat cgacctgtct cagctgggag gcgacgccta tccctatgac
    10321 gtgcccgatt atgccagcct gggcagcggc tcccccaaga aaaaacgcaa ggtggaagat
    10381 cctaagaaaa agcggaaagt ggacggcatt ggtagtggga gcaacggcag cagcggatcc
    10441 taagaattcc tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
    10501 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
    10561 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
    10621 gtggggtggg gcaggacagc aagggggagg attgggaaga gaatagcagg catgctgggg
    10681 agcggccgca ctagtctagt ctagagcgat cgcccgggtt taaacgctag ccaacaagct
    10741 cgtcatcgct ttgcagaaga gcagagagga tatgctcatc gtctaaagaa ctacccattt
    10801 tattatatat tagtcacgat atctataaca agaaaatata tatataataa gttatcacgt
    10861 aagtagaaca tgaaataaca atataattat cgtatgagtt aaatcttaaa agtcacgtaa
    10921 aagataatca tgcgtcattt tgactcacgc ggtcgttata gttcaaaatc agtgacactt
    10981 accgcattga caagcacgcc tcacgggagc tccaagcggc gactgagatg tcctaaatgc
    11041 acagcgacgg attcgcgcta tttagaaaga gagagcaata tttcaagaat gcatgcgtca
    11101 attttacgca gactatcttt ctagggttaa aaaagatttg cgctttactc gacctaaact
    11161 ttaaacacgt catagaatct tcgtttgaca aaaaccacat tgtggccaag ctgtgtgacg
    11221 cgacgcgcgc taaagaatgg caaaccaagt cgcgcgaggt accagctttt gttcccttta
    11281 gtgagggtta atttcgagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
    11341 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
    11401 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
    11461 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
    11521 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
    11581 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
    11641 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
    11701 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
    11761 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
    11821 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
    11881 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
    11941 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
    12001 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
    12061 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
    12121 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
    12181 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
    12241 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
    12301 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
    12361 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
    12421 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
    12481 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
    12541 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
    12601 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
    12661 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
    12721 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
    12781 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
    12841 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
    12901 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
    12961 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
    13021 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
    13081 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
    13141 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
    13201 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
    13261 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
    13321 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
    13381 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
    13441 cacatttccc cgaaaagtgc cacct
//
