LOCUS       sequence-194712-       14588 bp    DNA     circular SYN 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      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14588)
  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 14588)
  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..14588
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        24..128
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(129..197,198..989)
                     /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(347..366)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1160..1748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1649..1668
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1902..1919
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2036..2057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2072..2077,2078..2095,2096..2102)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2110..2126
                     /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     2134..2150
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2169..2189
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2171..2189
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   2240..2302
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        2540..2843
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2845..3122
                     /label=chicken beta-actin promoter
     intron          3123..4139
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     4063..4085
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     4147..4166
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             4206..4682
                     /codon_start=1
                     /gene="folA"
                     /product="E. coli dihydrofolate reductase"
                     /label=ecDHFR
                     /translation="MISLIAALAVDYVIGMENAMPWNLPADLAWFKRNTLNKPVIMGRH
                     TWESIGRPLPGRKNIILSSQPSTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVIEQFL
                     PKAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR"
     CDS             4707..4733
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4737..4757
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(4767..4790,4791..4793,4794..7349,7350..7352,
                     7353..8867)
                     /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(4767..4790,4791..4793,4794..7280,7281..7283,
                     7284..8867)
                     /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             8871..8891
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8928..9038
                     /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             9090..9200
                     /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             9252..9362
                     /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             9414..9524
                     /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             9573..10115
                     /codon_start=1
                     /product="C-terminal portion of the p65 subunit of mouse 
                     NF-kappa-B"
                     /label=p65
                     /translation="PSGQISNQALALAPSSAPVLAQTMVPSSAMVPLAQPPAPAPVLTP
                     GPPQSLSAPVPKSTQAGEGTLSEALLHLQFDADEDLGALLGNSTDPGVFTDLASVDNSE
                     FQQLLNQGVSMSHSTAEPMLMEYPEAITRLVTGSQRPPDPAPTPLGTSGLPNGLSGDED
                     FSSIADMDFSALLSQISS"
     CDS             10140..10511
                     /codon_start=1
                     /product="C-terminal activation domain from the human heat 
                     shock transcription factor HSF1"
                     /label=HSF1
                     /translation="GFSVDTSALLDLFSPSVTVPDMSLPDLDSSLASIQELLSPQEPPR
                     PPEAENSSPDSGKQLVHYTAQPLFLLDPGSVDTGSNDLPVLFELGEGSYFSEGDGFAED
                     PTISLLTGSEPPKAKDPTVS"
     CDS             10533..10586
                     /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(10593..10595,10596..10598,10599..11312)
                     /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(10593..10595,10596..10598,10599..11309)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(10899..10918)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     11246..11267
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    join(11334..11908,11909..11911,11912..11920)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(11501..11518)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     11728..11747
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             11921..12715
                     /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(11975..11994)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     12585..12604
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(13017..13034)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    13023..13247
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(13294..13310)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    13318..13334
                     /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(13342..13348,13349..13366,13367..13372))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13387..13408
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    complement(13467..13591)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     repeat_region   13881..13915
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(13944..13963)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(13945..13963)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(13973..13990)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(13973..13989)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13982..14004)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      14131..14586
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(14218..14237)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     14428..14449
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
       61 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
      121 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
      181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
      241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
      301 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
      361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
      421 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
      481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
      541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
      601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
      661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
      721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
      781 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
      841 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
      901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
      961 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     1021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     1081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     1321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     1561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     1621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     1681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     1741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     1801 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     1861 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     1921 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     1981 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     2041 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     2101 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     2161 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggattaatta agcggccgca
     2221 tacgcgtata ctagattaac cctagaaaga taatcatatt gtgacgtacg ttaaagataa
     2281 tcatgcgtaa aattgacgca tgtgttttat cggtctgtat atcgaggttt atttattaat
     2341 ttgaatagat attaagtttt attatattta cacttacata ctaataataa attcaacaaa
     2401 caatttattt atgtttattt atttattaaa aaaaaacaaa aactcaaaat ttcttctata
     2461 aagtaacaaa acttttaact agttattaat agtaatcaat tacggggtca ttagttcata
     2521 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     2581 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     2641 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     2701 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     2761 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     2821 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
     2881 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
     2941 cgatgggggc gggggggggg ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc
     3001 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
     3061 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
     3121 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
     3181 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
     3241 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
     3301 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
     3361 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
     3421 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     3481 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     3541 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     3601 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     3661 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     3721 cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg
     3781 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
     3841 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     3901 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     3961 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     4021 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     4081 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcntacagc
     4141 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttgggatccg
     4201 ccaccatgat cagtctgatt gcggcgttag cggtagatta cgttatcggc atggaaaacg
     4261 ccatgccgtg gaacctgcct gccgatctcg cctggtttaa acgcaacacc ttaaataaac
     4321 ccgtgattat gggccgccat acctgggaat caatcggtcg tccgttgcca ggacgcaaaa
     4381 atattatcct cagcagtcaa ccgagtacgg acgatcgcgt aacgtgggtg aagtcggtgg
     4441 atgaagccat cgcggcgtgt ggtgacgtac cagaaatcat ggtgattggc ggcggtcgcg
     4501 ttattgaaca gttcttgcca aaagcgcaaa aactgtatct gacgcatatc gacgcagaag
     4561 tggaaggcga cacccatttc ccggattacg agccggatga ctgggaatcg gtattcagcg
     4621 aattccacga tgctgatgcg cagaactctc acagctattg ctttgagatt ctggagcggc
     4681 gaggctccgg aaccggtgcc accatgtacc catacgatgt tccagattac gcttcgccga
     4741 agaaaaagcg caaggtcgaa gcgtccgaca agaagtacag catcggcctg gccatcggca
     4801 ccaactctgt gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc aagaaattca
     4861 aggtgctggg caacaccgac cggcacagca tcaagaagaa cctgatcgga gccctgctgt
     4921 tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag aaccgccaga agaagataca
     4981 ccagacggaa gaaccggatc tgctatctgc aagagatctt cagcaacgag atggccaagg
     5041 tggacgacag cttcttccac agactggaag agtccttcct ggtggaagag gataagaagc
     5101 acgagcggca ccccatcttc ggcaacatcg tggacgaggt ggcctaccac gagaagtacc
     5161 ccaccatcta ccacctgaga aagaaactgg tggacagcac cgacaaggcc gacctgcggc
     5221 tgatctatct ggccctggcc cacatgatca agttccgggg ccacttcctg atcgagggcg
     5281 acctgaaccc cgacaacagc gacgtggaca agctgttcat ccagctggtg cagacctaca
     5341 accagctgtt cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag gccatcctgt
     5401 ctgccagact gagcaagagc agacggctgg aaaatctgat cgcccagctg cccggcgaga
     5461 agaagaatgg cctgttcggc aacctgattg ccctgagcct gggcctgacc cccaacttca
     5521 agagcaactt cgacctggcc gaggatgcca aactgcagct gagcaaggac acctacgacg
     5581 acgacctgga caacctgctg gcccagatcg gcgaccagta cgccgacctg tttctggccg
     5641 ccaagaacct gtccgacgcc atcctgctga gcgacatcct gagagtgaac accgagatca
     5701 ccaaggcccc cctgagcgcc tctatgatca agagatacga cgagcaccac caggacctga
     5761 ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa gtacaaagag attttcttcg
     5821 accagagcaa gaacggctac gccggctaca ttgacggcgg agccagccag gaagagttct
     5881 acaagttcat caagcccatc ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc
     5941 tgaacagaga ggacctgctg cggaagcagc ggaccttcga caacggcagc atcccccacc
     6001 agatccacct gggagagctg cacgccattc tgcggcggca ggaagatttt tacccattcc
     6061 tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc tactacgtgg
     6121 gccctctggc caggggaaac agcagattcg cctggatgac cagaaagagc gaggaaacca
     6181 tcaccccctg gaacttcgag gaagtggtgg acaagggcgc ttccgcccag agcttcatcg
     6241 agcggatgac caacttcgat aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc
     6301 tgctgtacga gtacttcacc gtgtataacg agctgaccaa agtgaaatac gtgaccgagg
     6361 gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa ggccatcgtg gacctgctgt
     6421 tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga ggactacttc aagaaaatcg
     6481 agtgcttcga ctccgtggaa atctccggcg tggaagatcg gttcaacgcc tccctgggca
     6541 cataccacga tctgctgaaa attatcaagg acaaggactt cctggacaat gaggaaaacg
     6601 aggacattct ggaagatatc gtgctgaccc tgacactgtt tgaggacaga gagatgatcg
     6661 aggaacggct gaaaacctat gcccacctgt tcgacgacaa agtgatgaag cagctgaagc
     6721 ggcggagata caccggctgg ggcaggctga gccggaagct gatcaacggc atccgggaca
     6781 agcagtccgg caagacaatc ctggatttcc tgaagtccga cggcttcgcc aacagaaact
     6841 tcatgcagct gatccacgac gacagcctga cctttaaaga ggacatccag aaagcccagg
     6901 tgtccggcca gggcgatagc ctgcacgagc acattgccaa tctggccggc agccccgcca
     6961 ttaagaaggg catcctgcag acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc
     7021 ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga gaaccagacc acccagaagg
     7081 gacagaagaa cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa gagctgggca
     7141 gccagatcct gaaagaacac cccgtggaaa acacccagct gcagaacgag aagctgtacc
     7201 tgtactacct gcagaatggg cgggatatgt acgtggacca ggaactggac atcaaccggc
     7261 tgtccgacta cgatgtcgac gccatcgtgc ctcagagctt tctgaaggac gactccatcg
     7321 acaacaaggt gctgaccaga agcgacaaga accggggcaa gagcgacaac gtgccctccg
     7381 aagaggtcgt gaagaagatg aagaactact ggcggcagct gctgaacgcc aagctgatta
     7441 cccagagaaa gttcgacaat ctgaccaagg ccgagagagg cggcctgagc gaactggata
     7501 aggccggctt catcaagaga cagctggtgg aaacccggca gatcacaaag cacgtggcac
     7561 agatcctgga ctcccggatg aacactaagt acgacgagaa tgacaagctg atccgggaag
     7621 tgaaagtgat caccctgaag tccaagctgg tgtccgattt ccggaaggat ttccagtttt
     7681 acaaagtgcg cgagatcaac aactaccacc acgcccacga cgcctacctg aacgccgtcg
     7741 tgggaaccgc cctgatcaaa aagtacccta agctggaaag cgagttcgtg tacggcgact
     7801 acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta
     7861 ccgccaagta cttcttctac agcaacatca tgaacttttt caagaccgag attaccctgg
     7921 ccaacggcga gatccggaag cggcctctga tcgagacaaa cggcgaaacc ggggagatcg
     7981 tgtgggataa gggccgggat tttgccaccg tgcggaaagt gctgagcatg ccccaagtga
     8041 atatcgtgaa aaagaccgag gtgcagacag gcggcttcag caaagagtct atcctgccca
     8101 agaggaacag cgataagctg atcgccagaa agaaggactg ggaccctaag aagtacggcg
     8161 gcttcgacag ccccaccgtg gcctattctg tgctggtggt ggccaaagtg gaaaagggca
     8221 agtccaagaa actgaagagt gtgaaagagc tgctggggat caccatcatg gaaagaagca
     8281 gcttcgagaa gaatcccatc gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg
     8341 acctgatcat caagctgcct aagtactccc tgttcgagct ggaaaacggc cggaagagaa
     8401 tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact ggccctgccc tccaaatatg
     8461 tgaacttcct gtacctggcc agccactatg agaagctgaa gggctccccc gaggataatg
     8521 agcagaaaca gctgtttgtg gaacagcaca agcactacct ggacgagatc atcgagcaga
     8581 tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa tctggacaaa gtgctgtccg
     8641 cctacaacaa gcaccgggat aagcccatca gagagcaggc cgagaatatc atccacctgt
     8701 ttaccctgac caatctggga gcccctgccg ccttcaagta ctttgacacc accatcgacc
     8761 ggaagaggta caccagcacc aaagaggtgc tggacgccac cctgatccac cagagcatca
     8821 ccggcctgta cgagacacgg atcgacctgt ctcagctggg aggcgacagc cccaagaaga
     8881 agagaaaggt ggaggccagc ctcgagtgcg aaagcggcgg gccggccgac gcccttgacg
     8941 attttgactt agacatgctc ccagccgatg cccttgacga ctttgacctt gatatgctgc
     9001 ctgctgacgc tcttgacgat tttgaccttg acatgctccc cgggtcacta cccagcgcta
     9061 gcgtcgagtg cgaaagcggc gggccggccg acgcccttga cgattttgac ttagacatgc
     9121 tcccagccga tgcccttgac gactttgacc ttgatatgct gcctgctgac gctcttgacg
     9181 attttgacct tgacatgctc cccgggtcac tacccagcgc tagcgtcgag tgcgaaagcg
     9241 gcgggccggc cgacgccctt gacgattttg acttagacat gctcccagcc gatgcccttg
     9301 acgactttga ccttgatatg ctgcctgctg acgctcttga cgattttgac cttgacatgc
     9361 tccccgggtc actacccagc gctagcgtcg agtgcgaaag cggcgggccg gccgacgccc
     9421 ttgacgattt tgacttagac atgctcccag ccgatgccct tgacgacttt gaccttgata
     9481 tgctgcctgc tgacgctctt gacgattttg accttgacat gctccccggg tcactaccca
     9541 gcgctagcgt cgagttcgaa ggcagcggcg ggccttcagg gcagatcagc aaccaggccc
     9601 tggctctggc ccctagctcc gctccagtgc tggcccagac tatggtgccc tctagtgcta
     9661 tggtgcctct ggcccagcca cctgctccag cccctgtgct gaccccagga ccaccccagt
     9721 cactgagcgc tccagtgccc aagtctacac aggccggcga ggggactctg agtgaagctc
     9781 tgctgcacct gcagttcgac gctgatgagg acctgggagc tctgctgggg aacagcaccg
     9841 atcccggagt gttcacagat ctggcctccg tggacaactc tgagtttcag cagctgctga
     9901 atcagggcgt gtccatgtct catagtacag ccgaaccaat gctgatggag taccccgaag
     9961 ccattacccg gctggtgacc ggcagccagc ggccccccga ccccgctcca actcccctgg
    10021 gaaccagcgg cctgcctaat gggctgtccg gagatgaaga cttctcaagc atcgctgata
    10081 tggactttag tgccctgctg tcacagattt cctctagtgg gcagggagga ggtggaagcg
    10141 gcttcagcgt ggacaccagt gccctgctgg acctgttcag cccctcggtg accgtgcccg
    10201 acatgagcct gcctgacctt gacagcagcc tggccagtat ccaagagctc ctgtctcccc
    10261 aggagccccc caggcctccc gaggcagaga acagcagccc ggattcaggg aagcagctgg
    10321 tgcactacac agcgcagccg ctgttcctgc tggaccccgg ctccgtggac accgggagca
    10381 acgacctgcc ggtgctgttt gagctgggag agggctccta cttctccgaa ggggacggct
    10441 tcgccgagga ccccaccatc tccctgctga caggctcgga gcctcccaaa gccaaggacc
    10501 ccactgtctc ctctagcgtc gacggctccg gagagggcag aggaagtctg ctaacatgcg
    10561 gtgacgtcga ggagaatcct ggcccaatcg atatggtgag caagggcgag gagctgttca
    10621 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
    10681 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
    10741 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
    10801 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
    10861 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
    10921 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
    10981 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
    11041 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
    11101 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
    11161 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
    11221 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
    11281 tcactctcgg catggacgag ctgtacaagt aatttaaaga attccgcccc ccccccctct
    11341 ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt
    11401 gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct
    11461 ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa
    11521 ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg
    11581 tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc
    11641 caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg
    11701 agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg
    11761 aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc
    11821 tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg
    11881 gttttccttt gaaaaacacg atgataagct tgccacaacc atggctgaac aagatggatt
    11941 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
    12001 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
    12061 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
    12121 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
    12181 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
    12241 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
    12301 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
    12361 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
    12421 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
    12481 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
    12541 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
    12601 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
    12661 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
    12721 atcggctaga gtcgacccgg acgggacgct cctgcgcctg atacagaacg aattgcttgc
    12781 aggcatctca tgagtgtgtc ttcccgtttt ccgcctgagg tcactgcgtg gatggagcgc
    12841 tggcgcctgc tgcgcgacgg cgagctgctc accacccact cgagggctgc agcgctgcag
    12901 aggccgagtg cagaactgct ccaaagggac ctcaaggctt tccgagggac actaggctga
    12961 ctccatcgag ccagtgtaga gataagctta tcgatgatct agagctcgct gatcagcctc
    13021 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
    13081 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
    13141 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
    13201 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
    13261 aagaacctgc agcccaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
    13321 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
    13381 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
    13441 gggaaacctg tcgtgccagc ggatcccata gtgactggat atgttgtgtt ttacagtatt
    13501 atgtagtctg ttttttatgc aaaatctaat ttaatatatt gatatttata tcattttacg
    13561 tttctcgttc agcttttttg tacaaacttg tgctagcatt ctagttgata tctataacaa
    13621 gaaaatatat atataataag ttatcacgta agtagaacat gaaataacaa tataattatc
    13681 gtatgagtta aatcttaaaa gtcacgtaaa agataatcat gcgtcatttt gactcacgcg
    13741 gtcgttatag ttcaaaatca gtgacactta ccgcattgac aagcacgcct cacgggagct
    13801 ccaagcggcg actgagatgt cctaaatgca cagcgacgga ttcgcgctat ttagaaagag
    13861 agagcaatat ttcaagaatg catgcgtcaa ttttacgcag actatctttc tagggttaat
    13921 ctagaatggc gcgccccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg
    13981 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
    14041 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
    14101 aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg
    14161 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
    14221 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
    14281 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
    14341 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
    14401 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
    14461 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
    14521 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
    14581 caatttag
//
