LOCUS       sequence-259794-       13431 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac transposon system construct with constitutive CAG promoter 
            expression of TMP inducible DDdCas9VP192 activator followed by 
            T2A-EGFP as a reporter and IRES-Neo as selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13431)
  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 13431)
  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-DDdCas9VP192-T2A-GFP-IRES-Neo
FEATURES             Location/Qualifiers
     source          1..13431
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          50..1066
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     990..1012
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1074..1093
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1133..1609
                     /codon_start=1
                     /gene="folA"
                     /product="E. coli dihydrofolate reductase"
                     /label=ecDHFR
                     /translation="MISLIAALAVDYVIGMENAMPWNLPADLAWFKRNTLNKPVIMGRH
                     TWESIGRPLPGRKNIILSSQPSTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVIEQFL
                     PKAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR"
     CDS             1634..1660
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1664..1684
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1694..1717,1718..1720,1721..4276,4277..4279,
                     4280..5794)
                     /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(1694..1717,1718..1720,1721..4207,4208..4210,
                     4211..5794)
                     /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             5798..5818
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5855..5965
                     /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             6017..6127
                     /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             6179..6289
                     /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             6341..6451
                     /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             6491..6544
                     /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(6551..6553,6554..6556,6557..7270)
                     /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(6551..6553,6554..6556,6557..7267)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6857..6876)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7204..7225
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    7306..7873
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7467..7484)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7694..7713
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             7887..8681
                     /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(7941..7960)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8551..8570
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(8983..9000)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8989..9213
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(9260..9276)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    9284..9300
                     /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(9308..9314,9315..9332,9333..9338))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9353..9374
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    complement(9433..9557)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     repeat_region   9847..9881
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(9910..9929)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9911..9929)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9939..9956)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9939..9955)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9948..9970)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      10097..10552
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10184..10203)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10394..10415
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10578..10682
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10683..10751,10752..11543)
                     /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(10901..10920)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11714..12302
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12203..12222
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12456..12473
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12590..12611
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12626..12631,12632..12649,12650..12656)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12664..12680
                     /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     12688..12704
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12723..12743
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        12725..12743
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   12794..12856
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        13094..13397
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ccatckccss cssckscssc cctataaaaa gcgawgcgcg cggcgggcgg gagtcgctgc
       61 gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc cccggctctg
      121 actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa
      181 ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa gccttgaggg
      241 gctccgggag ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg tgtgtgtgtg
      301 cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct gcgggcgcgg
      361 cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg gcggtgcccc
      421 gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt gcgtgggggg
      481 gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc cccctccccg
      541 agttgctgag cacggcccgg cttcgggtgc ggggctccgt acggggcgtg gcgcggggct
      601 cgccgtgccg ggcggggggt ggcggcaggk gggggtgccg ggcggggcgg ggccgcctcg
      661 ggccggggrg ggctcggggg rggggcgcgg cggcccccgg agcgccggcg gctgtcgagg
      721 cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc agggacttcc
      781 tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc cctctagcgg
      841 gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc
      901 gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc ggggggacgg
      961 ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga ccggcggctc
     1021 tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc tgggcaacgt
     1081 gctggttatt gtgctgtctc atcattttgg caaagaattg ggatccgcca ccatgatcag
     1141 tctgattgcg gcgttagcgg tagattacgt tatcggcatg gaaaacgcca tgccgtggaa
     1201 cctgcctgcc gatctcgcct ggtttaaacg caacacctta aataaacccg tgattatggg
     1261 ccgccatacc tgggaatcaa tcggtcgtcc gttgccagga cgcaaaaata ttatcctcag
     1321 cagtcaaccg agtacggacg atcgcgtaac gtgggtgaag tcggtggatg aagccatcgc
     1381 ggcgtgtggt gacgtaccag aaatcatggt gattggcggc ggtcgcgtta ttgaacagtt
     1441 cttgccaaaa gcgcaaaaac tgtatctgac gcatatcgac gcagaagtgg aaggcgacac
     1501 ccatttcccg gattacgagc cggatgactg ggaatcggta ttcagcgaat tccacgatgc
     1561 tgatgcgcag aactctcaca gctattgctt tgagattctg gagcggcgag gctccggaac
     1621 cggtgccacc atgtacccat acgatgttcc agattacgct tcgccgaaga aaaagcgcaa
     1681 ggtcgaagcg tccgacaaga agtacagcat cggcctggcc atcggcacca actctgtggg
     1741 ctgggccgtg atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa
     1801 caccgaccgg cacagcatca agaagaacct gatcggagcc ctgctgttcg acagcggcga
     1861 aacagccgag gccacccggc tgaagagaac cgccagaaga agatacacca gacggaagaa
     1921 ccggatctgc tatctgcaag agatcttcag caacgagatg gccaaggtgg acgacagctt
     1981 cttccacaga ctggaagagt ccttcctggt ggaagaggat aagaagcacg agcggcaccc
     2041 catcttcggc aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca
     2101 cctgagaaag aaactggtgg acagcaccga caaggccgac ctgcggctga tctatctggc
     2161 cctggcccac atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga
     2221 caacagcgac gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga
     2281 ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg ccagactgag
     2341 caagagcaga cggctggaaa atctgatcgc ccagctgccc ggcgagaaga agaatggcct
     2401 gttcggcaac ctgattgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga
     2461 cctggccgag gatgccaaac tgcagctgag caaggacacc tacgacgacg acctggacaa
     2521 cctgctggcc cagatcggcg accagtacgc cgacctgttt ctggccgcca agaacctgtc
     2581 cgacgccatc ctgctgagcg acatcctgag agtgaacacc gagatcacca aggcccccct
     2641 gagcgcctct atgatcaaga gatacgacga gcaccaccag gacctgaccc tgctgaaagc
     2701 tctcgtgcgg cagcagctgc ctgagaagta caaagagatt ttcttcgacc agagcaagaa
     2761 cggctacgcc ggctacattg acggcggagc cagccaggaa gagttctaca agttcatcaa
     2821 gcccatcctg gaaaagatgg acggcaccga ggaactgctc gtgaagctga acagagagga
     2881 cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg
     2941 agagctgcac gccattctgc ggcggcagga agatttttac ccattcctga aggacaaccg
     3001 ggaaaagatc gagaagatcc tgaccttccg catcccctac tacgtgggcc ctctggccag
     3061 gggaaacagc agattcgcct ggatgaccag aaagagcgag gaaaccatca ccccctggaa
     3121 cttcgaggaa gtggtggaca agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa
     3181 cttcgataag aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta
     3241 cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc
     3301 cgccttcctg agcggcgagc agaaaaaggc catcgtggac ctgctgttca agaccaaccg
     3361 gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt gcttcgactc
     3421 cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc ctgggcacat accacgatct
     3481 gctgaaaatt atcaaggaca aggacttcct ggacaatgag gaaaacgagg acattctgga
     3541 agatatcgtg ctgaccctga cactgtttga ggacagagag atgatcgagg aacggctgaa
     3601 aacctatgcc cacctgttcg acgacaaagt gatgaagcag ctgaagcggc ggagatacac
     3661 cggctggggc aggctgagcc ggaagctgat caacggcatc cgggacaagc agtccggcaa
     3721 gacaatcctg gatttcctga agtccgacgg cttcgccaac agaaacttca tgcagctgat
     3781 ccacgacgac agcctgacct ttaaagagga catccagaaa gcccaggtgt ccggccaggg
     3841 cgatagcctg cacgagcaca ttgccaatct ggccggcagc cccgccatta agaagggcat
     3901 cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc acaagcccga
     3961 gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc cagaagggac agaagaacag
     4021 ccgcgagaga atgaagcgga tcgaagaggg catcaaagag ctgggcagcc agatcctgaa
     4081 agaacacccc gtggaaaaca cccagctgca gaacgagaag ctgtacctgt actacctgca
     4141 gaatgggcgg gatatgtacg tggaccagga actggacatc aaccggctgt ccgactacga
     4201 tgtcgacgcc atcgtgcctc agagctttct gaaggacgac tccatcgaca acaaggtgct
     4261 gaccagaagc gacaagaacc ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa
     4321 gaagatgaag aactactggc ggcagctgct gaacgccaag ctgattaccc agagaaagtt
     4381 cgacaatctg accaaggccg agagaggcgg cctgagcgaa ctggataagg ccggcttcat
     4441 caagagacag ctggtggaaa cccggcagat cacaaagcac gtggcacaga tcctggactc
     4501 ccggatgaac actaagtacg acgagaatga caagctgatc cgggaagtga aagtgatcac
     4561 cctgaagtcc aagctggtgt ccgatttccg gaaggatttc cagttttaca aagtgcgcga
     4621 gatcaacaac taccaccacg cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct
     4681 gatcaaaaag taccctaagc tggaaagcga gttcgtgtac ggcgactaca aggtgtacga
     4741 cgtgcggaag atgatcgcca agagcgagca ggaaatcggc aaggctaccg ccaagtactt
     4801 cttctacagc aacatcatga actttttcaa gaccgagatt accctggcca acggcgagat
     4861 ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt gggataaggg
     4921 ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa
     4981 gaccgaggtg cagacaggcg gcttcagcaa agagtctatc ctgcccaaga ggaacagcga
     5041 taagctgatc gccagaaaga aggactggga ccctaagaag tacggcggct tcgacagccc
     5101 caccgtggcc tattctgtgc tggtggtggc caaagtggaa aagggcaagt ccaagaaact
     5161 gaagagtgtg aaagagctgc tggggatcac catcatggaa agaagcagct tcgagaagaa
     5221 tcccatcgac tttctggaag ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa
     5281 gctgcctaag tactccctgt tcgagctgga aaacggccgg aagagaatgc tggcctctgc
     5341 cggcgaactg cagaagggaa acgaactggc cctgccctcc aaatatgtga acttcctgta
     5401 cctggccagc cactatgaga agctgaaggg ctcccccgag gataatgagc agaaacagct
     5461 gtttgtggaa cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttctc
     5521 caagagagtg atcctggccg acgctaatct ggacaaagtg ctgtccgcct acaacaagca
     5581 ccgggataag cccatcagag agcaggccga gaatatcatc cacctgttta ccctgaccaa
     5641 tctgggagcc cctgccgcct tcaagtactt tgacaccacc atcgaccgga agaggtacac
     5701 cagcaccaaa gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga
     5761 gacacggatc gacctgtctc agctgggagg cgacagcccc aagaagaaga gaaaggtgga
     5821 ggccagcctc gagtgcgaaa gcggcgggcc ggccgacgcc cttgacgatt ttgacttaga
     5881 catgctccca gccgatgccc ttgacgactt tgaccttgat atgctgcctg ctgacgctct
     5941 tgacgatttt gaccttgaca tgctccccgg gtcactaccc agcgctagcg tcgagtgcga
     6001 aagcggcggg ccggccgacg cccttgacga ttttgactta gacatgctcc cagccgatgc
     6061 ccttgacgac tttgaccttg atatgctgcc tgctgacgct cttgacgatt ttgaccttga
     6121 catgctcccc gggtcactac ccagcgctag cgtcgagtgc gaaagcggcg ggccggccga
     6181 cgcccttgac gattttgact tagacatgct cccagccgat gcccttgacg actttgacct
     6241 tgatatgctg cctgctgacg ctcttgacga ttttgacctt gacatgctcc ccgggtcact
     6301 acccagcgct agcgtcgagt gcgaaagcgg cgggccggcc gacgcccttg acgattttga
     6361 cttagacatg ctcccagccg atgcccttga cgactttgac cttgatatgc tgcctgctga
     6421 cgctcttgac gattttgacc ttgacatgct ccccgggtca ctacccagcg ctagcgtcga
     6481 cggctccgga gagggcagag gaagtctgct aacatgcggt gacgtcgagg agaatcctgg
     6541 cccaatcgat atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     6601 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     6661 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     6721 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     6781 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     6841 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     6901 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     6961 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     7021 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     7081 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     7141 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     7201 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     7261 gtacaagtaa tttaaagaat tccgccccyc wcbmssckms mmstctccck cccccccccc
     7321 taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt
     7381 ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt
     7441 gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt
     7501 cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct
     7561 ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt
     7621 ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt
     7681 ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa
     7741 ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta
     7801 gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa
     7861 aacacgatga taagcttgcc acaaccatgg ctgaacaaga tggattgcac gcaggttctc
     7921 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     7981 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     8041 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     8101 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     8161 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     8221 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     8281 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     8341 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     8401 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     8461 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     8521 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     8581 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     8641 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggatcg gctagagtcg
     8701 acccggacgg gacgctcctg cgcctgatac agaacgaatt gcttgcaggc atctcatgag
     8761 tgtgtcttcc cgttttccgc ctgaggtcac tgcgtggatg gagcgctggc gcctgctgcg
     8821 cgacggcgag ctgctcacca cccactcgag ggctgcagcg ctgcagaggc cgagtgcaga
     8881 actgctccaa agggacctca aggctttccg agggacacta ggctgactcc atcgagccag
     8941 tgtagagata agcttatcga tgatctagag ctcgctgatc agcctcgact gtgccttcta
     9001 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     9061 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     9121 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     9181 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga acctgcagcc
     9241 caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     9301 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     9361 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     9421 gccagcggat cccatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt
     9481 ttatgcaaaa tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcagct
     9541 tttttgtaca aacttgtgct agcattctag ttgatatcta taacaagaaa atatatatat
     9601 aataagttat cacgtaagta gaacatgaaa taacaatata attatcgtat gagttaaatc
     9661 ttaaaagtca cgtaaaagat aatcatgcgt cattttgact cacgcggtcg ttatagttca
     9721 aaatcagtga cacttaccgc attgacaagc acgcctcacg ggagctccaa gcggcgactg
     9781 agatgtccta aatgcacagc gacggattcg cgctatttag aaagagagag caatatttca
     9841 agaatgcatg cgtcaatttt acgcagacta tctttctagg gttaatctag aatggcgcgc
     9901 cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt
     9961 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
    10021 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
    10081 ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
    10141 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
    10201 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
    10261 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
    10321 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
    10381 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
    10441 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
    10501 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
    10561 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
    10621 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
    10681 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
    10741 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
    10801 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
    10861 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
    10921 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
    10981 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
    11041 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
    11101 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
    11161 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
    11221 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
    11281 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
    11341 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
    11401 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
    11461 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
    11521 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
    11581 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
    11641 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
    11701 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
    11761 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
    11821 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
    11881 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
    11941 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
    12001 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
    12061 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
    12121 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
    12181 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
    12241 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
    12301 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
    12361 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
    12421 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
    12481 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
    12541 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
    12601 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
    12661 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
    12721 gcgcaattaa ccctcactaa agggaacaaa agctggatta attaagcggc cgcatacgcg
    12781 tatactagat taaccctaga aagataatca tattgtgacg tacgttaaag ataatcatgc
    12841 gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag gtttatttat taatttgaat
    12901 agatattaag ttttattata tttacactta catactaata ataaattcaa caaacaattt
    12961 atttatgttt atttatttat taaaaaaaaa caaaaactca aaatttcttc tataaagtaa
    13021 caaaactttt aactagttat taatagtaat caattacggg gtcattagtt catagcccat
    13081 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
    13141 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
    13201 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
    13261 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
    13321 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
    13381 tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc c
//
