LOCUS       sequence-198237-       13809 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  PiggyBac transposon system construct with constitutive CAG promoter 
            expression of TMP inducible DDdCas9VP192-p65-HSF1 activator followed
            by T2A-EGFP as a reporter and IRES-Neo as selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 13809)
  AUTHORS   Weltner J, Balboa D, Katayama S, Bespalov M, Krjutskov K, Jouhilahti
            EM, Trokovic R, Kere J, Otonkoski T
  TITLE     Human pluripotent reprogramming with CRISPR activators.
  JOURNAL   Nat Commun. 2018 Jul 6;9(1):2643. doi: 10.1038/s41467-018-05067-x.
  PUBMED    29980666
REFERENCE   2  (bases 1 to 13809)
  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: "2018-07-6"; volume: "9(1):2643. doi"; pages: " 
            10.1038/s41467-018-05067-x"
COMMENT     Sequence Label: PB-CAG-DDdCas9VPH-T2A-GFP-IRES-Neo
FEATURES             Location/Qualifiers
     source          1..13809
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     322..344
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     406..425
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             465..941
                     /codon_start=1
                     /gene="folA"
                     /product="E. coli dihydrofolate reductase"
                     /label=ecDHFR
                     /translation="MISLIAALAVDYVIGMENAMPWNLPADLAWFKRNTLNKPVIMGRH
                     TWESIGRPLPGRKNIILSSQPSTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVIEQFL
                     PKAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR"
     CDS             966..992
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             996..1016
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1026..1049,1050..1052,1053..3608,3609..3611,
                     3612..5126)
                     /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 N863A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(1026..1049,1050..1052,1053..3539,3540..3542,
                     3543..5126)
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5130..5150
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5187..5297
                     /codon_start=1
                     /product="3 tandem repeats of the minimal activation domain
                     of herpes simplex virus VP16 (Cheng et al., 2013)"
                     /label=VP48
                     /translation="DALDDFDLDMLPADALDDFDLDMLPADALDDFDLDML"
     CDS             5349..5459
                     /codon_start=1
                     /product="3 tandem repeats of the minimal activation domain
                     of herpes simplex virus VP16 (Cheng et al., 2013)"
                     /label=VP48
                     /translation="DALDDFDLDMLPADALDDFDLDMLPADALDDFDLDML"
     CDS             5511..5621
                     /codon_start=1
                     /product="3 tandem repeats of the minimal activation domain
                     of herpes simplex virus VP16 (Cheng et al., 2013)"
                     /label=VP48
                     /translation="DALDDFDLDMLPADALDDFDLDMLPADALDDFDLDML"
     CDS             5673..5783
                     /codon_start=1
                     /product="3 tandem repeats of the minimal activation domain
                     of herpes simplex virus VP16 (Cheng et al., 2013)"
                     /label=VP48
                     /translation="DALDDFDLDMLPADALDDFDLDMLPADALDDFDLDML"
     CDS             5832..6374
                     /codon_start=1
                     /product="C-terminal portion of the p65 subunit of mouse 
                     NF-kappa-B"
                     /label=p65
                     /translation="PSGQISNQALALAPSSAPVLAQTMVPSSAMVPLAQPPAPAPVLTP
                     GPPQSLSAPVPKSTQAGEGTLSEALLHLQFDADEDLGALLGNSTDPGVFTDLASVDNSE
                     FQQLLNQGVSMSHSTAEPMLMEYPEAITRLVTGSQRPPDPAPTPLGTSGLPNGLSGDED
                     FSSIADMDFSALLSQISS"
     CDS             6399..6770
                     /codon_start=1
                     /product="C-terminal activation domain from the human heat 
                     shock transcription factor HSF1"
                     /label=HSF1
                     /translation="GFSVDTSALLDLFSPSVTVPDMSLPDLDSSLASIQELLSPQEPPR
                     PPEAENSSPDSGKQLVHYTAQPLFLLDPGSVDTGSNDLPVLFELGEGSYFSEGDGFAED
                     PTISLLTGSEPPKAKDPTVS"
     CDS             6792..6845
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(6852..6854,6855..6857,6858..7571)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(6852..6854,6855..6857,6858..7568)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7158..7177)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7505..7526
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    7593..8166
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7760..7777)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7987..8006
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             8180..8974
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(8234..8253)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8844..8863
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(9276..9293)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9282..9506
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(9553..9569)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    9577..9593
                     /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(9601..9607,9608..9625,9626..9631))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9646..9667
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    complement(9726..9850)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     repeat_region   10140..10174
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(10203..10222)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(10204..10222)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(10232..10249)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10232..10248)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10241..10263)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      10390..10845
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10477..10496)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10687..10708
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10871..10975
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10976..11044,11045..11836)
                     /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     complement(11194..11213)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      12007..12595
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12496..12515
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12749..12766
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12883..12904
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12919..12924,12925..12942,12943..12949)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12957..12973
                     /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)."
     primer_bind     12981..12997
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     13016..13036
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        13018..13036
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   13087..13149
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        13387..13690
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 agggctcggg ggaggggcgc ggcggccccc ggagcgccgg cggctgtcga ggcgcggcga
       61 gccgcagcca ttgcctttta tggtaatcgt gcgagagggc gcagggactt cctttgtccc
      121 aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac cccctctagc gggcgcgggg
      181 cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt gcgtcgccgc
      241 gccgccgtcc ccttctccct ctccagcctc ggggctgtcc gcggggggac ggctgccttc
      301 gggggggacg gggcagggcg gggttcggct tctggcgtgt gaccggcggc tctagagcct
      361 ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac gtgctggtta
      421 ttgtgctgtc tcatcatttt ggcaaagaat tgggatccgc caccatgatc agtctgattg
      481 cggcgttagc ggtagattac gttatcggca tggaaaacgc catgccgtgg aacctgcctg
      541 ccgatctcgc ctggtttaaa cgcaacacct taaataaacc cgtgattatg ggccgccata
      601 cctgggaatc aatcggtcgt ccgttgccag gacgcaaaaa tattatcctc agcagtcaac
      661 cgagtacgga cgatcgcgta acgtgggtga agtcggtgga tgaagccatc gcggcgtgtg
      721 gtgacgtacc agaaatcatg gtgattggcg gcggtcgcgt tattgaacag ttcttgccaa
      781 aagcgcaaaa actgtatctg acgcatatcg acgcagaagt ggaaggcgac acccatttcc
      841 cggattacga gccggatgac tgggaatcgg tattcagcga attccacgat gctgatgcgc
      901 agaactctca cagctattgc tttgagattc tggagcggcg aggctccgga accggtgcca
      961 ccatgtaccc atacgatgtt ccagattacg cttcgccgaa gaaaaagcgc aaggtcgaag
     1021 cgtccgacaa gaagtacagc atcggcctgg ccatcggcac caactctgtg ggctgggccg
     1081 tgatcaccga cgagtacaag gtgcccagca agaaattcaa ggtgctgggc aacaccgacc
     1141 ggcacagcat caagaagaac ctgatcggag ccctgctgtt cgacagcggc gaaacagccg
     1201 aggccacccg gctgaagaga accgccagaa gaagatacac cagacggaag aaccggatct
     1261 gctatctgca agagatcttc agcaacgaga tggccaaggt ggacgacagc ttcttccaca
     1321 gactggaaga gtccttcctg gtggaagagg ataagaagca cgagcggcac cccatcttcg
     1381 gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc caccatctac cacctgagaa
     1441 agaaactggt ggacagcacc gacaaggccg acctgcggct gatctatctg gccctggccc
     1501 acatgatcaa gttccggggc cacttcctga tcgagggcga cctgaacccc gacaacagcg
     1561 acgtggacaa gctgttcatc cagctggtgc agacctacaa ccagctgttc gaggaaaacc
     1621 ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc tgccagactg agcaagagca
     1681 gacggctgga aaatctgatc gcccagctgc ccggcgagaa gaagaatggc ctgttcggca
     1741 acctgattgc cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc gacctggccg
     1801 aggatgccaa actgcagctg agcaaggaca cctacgacga cgacctggac aacctgctgg
     1861 cccagatcgg cgaccagtac gccgacctgt ttctggccgc caagaacctg tccgacgcca
     1921 tcctgctgag cgacatcctg agagtgaaca ccgagatcac caaggccccc ctgagcgcct
     1981 ctatgatcaa gagatacgac gagcaccacc aggacctgac cctgctgaaa gctctcgtgc
     2041 ggcagcagct gcctgagaag tacaaagaga ttttcttcga ccagagcaag aacggctacg
     2101 ccggctacat tgacggcgga gccagccagg aagagttcta caagttcatc aagcccatcc
     2161 tggaaaagat ggacggcacc gaggaactgc tcgtgaagct gaacagagag gacctgctgc
     2221 ggaagcagcg gaccttcgac aacggcagca tcccccacca gatccacctg ggagagctgc
     2281 acgccattct gcggcggcag gaagattttt acccattcct gaaggacaac cgggaaaaga
     2341 tcgagaagat cctgaccttc cgcatcccct actacgtggg ccctctggcc aggggaaaca
     2401 gcagattcgc ctggatgacc agaaagagcg aggaaaccat caccccctgg aacttcgagg
     2461 aagtggtgga caagggcgct tccgcccaga gcttcatcga gcggatgacc aacttcgata
     2521 agaacctgcc caacgagaag gtgctgccca agcacagcct gctgtacgag tacttcaccg
     2581 tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc
     2641 tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt caagaccaac cggaaagtga
     2701 ccgtgaagca gctgaaagag gactacttca agaaaatcga gtgcttcgac tccgtggaaa
     2761 tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa
     2821 ttatcaagga caaggacttc ctggacaatg aggaaaacga ggacattctg gaagatatcg
     2881 tgctgaccct gacactgttt gaggacagag agatgatcga ggaacggctg aaaacctatg
     2941 cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg gcggagatac accggctggg
     3001 gcaggctgag ccggaagctg atcaacggca tccgggacaa gcagtccggc aagacaatcc
     3061 tggatttcct gaagtccgac ggcttcgcca acagaaactt catgcagctg atccacgacg
     3121 acagcctgac ctttaaagag gacatccaga aagcccaggt gtccggccag ggcgatagcc
     3181 tgcacgagca cattgccaat ctggccggca gccccgccat taagaagggc atcctgcaga
     3241 cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg
     3301 tgatcgaaat ggccagagag aaccagacca cccagaaggg acagaagaac agccgcgaga
     3361 gaatgaagcg gatcgaagag ggcatcaaag agctgggcag ccagatcctg aaagaacacc
     3421 ccgtggaaaa cacccagctg cagaacgaga agctgtacct gtactacctg cagaatgggc
     3481 gggatatgta cgtggaccag gaactggaca tcaaccggct gtccgactac gatgtcgacg
     3541 ccatcgtgcc tcagagcttt ctgaaggacg actccatcga caacaaggtg ctgaccagaa
     3601 gcgacaagaa ccggggcaag agcgacaacg tgccctccga agaggtcgtg aagaagatga
     3661 agaactactg gcggcagctg ctgaacgcca agctgattac ccagagaaag ttcgacaatc
     3721 tgaccaaggc cgagagaggc ggcctgagcg aactggataa ggccggcttc atcaagagac
     3781 agctggtgga aacccggcag atcacaaagc acgtggcaca gatcctggac tcccggatga
     3841 acactaagta cgacgagaat gacaagctga tccgggaagt gaaagtgatc accctgaagt
     3901 ccaagctggt gtccgatttc cggaaggatt tccagtttta caaagtgcgc gagatcaaca
     3961 actaccacca cgcccacgac gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa
     4021 agtaccctaa gctggaaagc gagttcgtgt acggcgacta caaggtgtac gacgtgcgga
     4081 agatgatcgc caagagcgag caggaaatcg gcaaggctac cgccaagtac ttcttctaca
     4141 gcaacatcat gaactttttc aagaccgaga ttaccctggc caacggcgag atccggaagc
     4201 ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt gtgggataag ggccgggatt
     4261 ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg
     4321 tgcagacagg cggcttcagc aaagagtcta tcctgcccaa gaggaacagc gataagctga
     4381 tcgccagaaa gaaggactgg gaccctaaga agtacggcgg cttcgacagc cccaccgtgg
     4441 cctattctgt gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg
     4501 tgaaagagct gctggggatc accatcatgg aaagaagcag cttcgagaag aatcccatcg
     4561 actttctgga agccaagggc tacaaagaag tgaaaaagga cctgatcatc aagctgccta
     4621 agtactccct gttcgagctg gaaaacggcc ggaagagaat gctggcctct gccggcgaac
     4681 tgcagaaggg aaacgaactg gccctgccct ccaaatatgt gaacttcctg tacctggcca
     4741 gccactatga gaagctgaag ggctcccccg aggataatga gcagaaacag ctgtttgtgg
     4801 aacagcacaa gcactacctg gacgagatca tcgagcagat cagcgagttc tccaagagag
     4861 tgatcctggc cgacgctaat ctggacaaag tgctgtccgc ctacaacaag caccgggata
     4921 agcccatcag agagcaggcc gagaatatca tccacctgtt taccctgacc aatctgggag
     4981 cccctgccgc cttcaagtac tttgacacca ccatcgaccg gaagaggtac accagcacca
     5041 aagaggtgct ggacgccacc ctgatccacc agagcatcac cggcctgtac gagacacgga
     5101 tcgacctgtc tcagctggga ggcgacagcc ccaagaagaa gagaaaggtg gaggccagcc
     5161 tcgagtgcga aagcggcggg ccggccgacg cccttgacga ttttgactta gacatgctcc
     5221 cagccgatgc ccttgacgac tttgaccttg atatgctgcc tgctgacgct cttgacgatt
     5281 ttgaccttga catgctcccc gggtcactac ccagcgctag cgtcgagtgc gaaagcggcg
     5341 ggccggccga cgcccttgac gattttgact tagacatgct cccagccgat gcccttgacg
     5401 actttgacct tgatatgctg cctgctgacg ctcttgacga ttttgacctt gacatgctcc
     5461 ccgggtcact acccagcgct agcgtcgagt gcgaaagcgg cgggccggcc gacgcccttg
     5521 acgattttga cttagacatg ctcccagccg atgcccttga cgactttgac cttgatatgc
     5581 tgcctgctga cgctcttgac gattttgacc ttgacatgct ccccgggtca ctacccagcg
     5641 ctagcgtcga gtgcgaaagc ggcgggccgg ccgacgccct tgacgatttt gacttagaca
     5701 tgctcccagc cgatgccctt gacgactttg accttgatat gctgcctgct gacgctcttg
     5761 acgattttga ccttgacatg ctccccgggt cactacccag cgctagcgtc gagttcgaag
     5821 gcagcggcgg gccttcaggg cagatcagca accaggccct ggctctggcc cctagctccg
     5881 ctccagtgct ggcccagact atggtgccct ctagtgctat ggtgcctctg gcccagccac
     5941 ctgctccagc ccctgtgctg accccaggac caccccagtc actgagcgct ccagtgccca
     6001 agtctacaca ggccggcgag gggactctga gtgaagctct gctgcacctg cagttcgacg
     6061 ctgatgagga cctgggagct ctgctgggga acagcaccga tcccggagtg ttcacagatc
     6121 tggcctccgt ggacaactct gagtttcagc agctgctgaa tcagggcgtg tccatgtctc
     6181 atagtacagc cgaaccaatg ctgatggagt accccgaagc cattacccgg ctggtgaccg
     6241 gcagccagcg gccccccgac cccgctccaa ctcccctggg aaccagcggc ctgcctaatg
     6301 ggctgtccgg agatgaagac ttctcaagca tcgctgatat ggactttagt gccctgctgt
     6361 cacagatttc ctctagtggg cagggaggag gtggaagcgg cttcagcgtg gacaccagtg
     6421 ccctgctgga cctgttcagc ccctcggtga ccgtgcccga catgagcctg cctgaccttg
     6481 acagcagcct ggccagtatc caagagctcc tgtctcccca ggagcccccc aggcctcccg
     6541 aggcagagaa cagcagcccg gattcaggga agcagctggt gcactacaca gcgcagccgc
     6601 tgttcctgct ggaccccggc tccgtggaca ccgggagcaa cgacctgccg gtgctgtttg
     6661 agctgggaga gggctcctac ttctccgaag gggacggctt cgccgaggac cccaccatct
     6721 ccctgctgac aggctcggag cctcccaaag ccaaggaccc cactgtctcc tctagcgtcg
     6781 acggctccgg agagggcaga ggaagtctgc taacatgcgg tgacgtcgag gagaatcctg
     6841 gcccaatcga tatggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     6901 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     6961 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     7021 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     7081 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     7141 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     7201 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     7261 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     7321 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     7381 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     7441 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     7501 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     7561 tgtacaagta atttaaagaa ttccgccccm cccccctctc nnnnnnnncc ccctaacgtt
     7621 actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc
     7681 atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc
     7741 attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag
     7801 gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg
     7861 cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat
     7921 acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga
     7981 gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc
     8041 cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg
     8101 ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga
     8161 tgataagctt gccacaacca tggctgaaca agatggattg cacgcaggtt ctccggccgc
     8221 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     8281 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     8341 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     8401 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     8461 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     8521 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     8581 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     8641 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     8701 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     8761 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     8821 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     8881 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     8941 cgccttctat cgccttcttg acgagttctt ctgagcggga tcggctagag tcgacccgga
     9001 cgggacgctc ctgcgcctga tacagaacga attgcttgca ggcatctcat gagtgtgtct
     9061 tcccgttttc cgcctgaggt cactgcgtgg atggagcgct ggcgcctgct gcgcgacggc
     9121 gagctgctca ccacccactc gagggctgca gcgctgcaga ggccgagtgc agaactgctc
     9181 caaagggacc tcaaggcttt ccgagggaca ctaggctgac tccatcgagc cagtgtagag
     9241 ataagcttat cgatgatcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca
     9301 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     9361 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     9421 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     9481 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaacctgca gcccaagctt
     9541 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     9601 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     9661 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg
     9721 gatcccatag tgactggata tgttgtgttt tacagtatta tgtagtctgt tttttatgca
     9781 aaatctaatt taatatattg atatttatat cattttacgt ttctcgttca gcttttttgt
     9841 acaaacttgt gctagcattc tagttgatat ctataacaag aaaatatata tataataagt
     9901 tatcacgtaa gtagaacatg aaataacaat ataattatcg tatgagttaa atcttaaaag
     9961 tcacgtaaaa gataatcatg cgtcattttg actcacgcgg tcgttatagt tcaaaatcag
    10021 tgacacttac cgcattgaca agcacgcctc acgggagctc caagcggcga ctgagatgtc
    10081 ctaaatgcac agcgacggat tcgcgctatt tagaaagaga gagcaatatt tcaagaatgc
    10141 atgcgtcaat tttacgcaga ctatctttct agggttaatc tagaatggcg cgccccaatt
    10201 cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact
    10261 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
    10321 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
    10381 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
    10441 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
    10501 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
    10561 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
    10621 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
    10681 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
    10741 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
    10801 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
    10861 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
    10921 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
    10981 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
    11041 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
    11101 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
    11161 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
    11221 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
    11281 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
    11341 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
    11401 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
    11461 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
    11521 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
    11581 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
    11641 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
    11701 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
    11761 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
    11821 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
    11881 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
    11941 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
    12001 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
    12061 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
    12121 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
    12181 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
    12241 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
    12301 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
    12361 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
    12421 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
    12481 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
    12541 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
    12601 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
    12661 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
    12721 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
    12781 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
    12841 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
    12901 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
    12961 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat
    13021 taaccctcac taaagggaac aaaagctgga ttaattaagc ggccgcatac gcgtatacta
    13081 gattaaccct agaaagataa tcatattgtg acgtacgtta aagataatca tgcgtaaaat
    13141 tgacgcatgt gttttatcgg tctgtatatc gaggtttatt tattaatttg aatagatatt
    13201 aagttttatt atatttacac ttacatacta ataataaatt caacaaacaa tttatttatg
    13261 tttatttatt tattaaaaaa aaacaaaaac tcaaaatttc ttctataaag taacaaaact
    13321 tttaactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
    13381 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
    13441 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
    13501 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
    13561 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
    13621 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
    13681 tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc
    13741 cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg
    13801 ggggggggg
//
