LOCUS       sequence-254082-        8034 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for LUMIER assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8034)
  AUTHORS   Kawano S, Maruyama J, Nagashima S, Inami K, Qiu W, Iwasa H, Nakagawa
            K, Ishigami-Yuasa M, Kagechika H, Nishina H, Hata Y
  TITLE     A cell-based screening for TAZ activators identifies ethacridine, a 
            widely used antiseptic and abortifacient, as a compound that 
            promotes dephosphorylation of TAZ and inhibits adipogenesis in 
            C3H10T1/2 cells.
  JOURNAL   J Biochem. 2015 May 14. pii: mvv051.
  PUBMED    25979969
REFERENCE   2  (bases 1 to 8034)
  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: "J Biochem. 
            2015 May 14. pii: mvv051."
COMMENT     Sequence Label: pCIneoLuc-PP2A
FEATURES             Location/Qualifiers
     source          1..8034
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..212
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     163..183
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     209..233
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          373..505
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     488..507
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        550..568
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      572..581
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             578..2227
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(644..662)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    3221..3342
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3232..3251)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3286..3305
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3528..3983
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3615..3634)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3825..3846
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4100..4457
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4100..4291
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4123..4143)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4308..4443
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4508..5302
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4562..4581)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5172..5191
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5366..5414
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5455..5474)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5574..5596
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5634..5652)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5720..5824
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5825..5893,5894..6685)
                     /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(6043..6062)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6856..7444
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7345..7364
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7655..8034
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctgcgatcgc ccgccccgtt gacgcaaatg ggcggtaggc
      181 gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag atcactagaa
      241 gctttattgc ggtagtttat cacagttaaa ttgctaacgc agtcagtgct tctgacacaa
      301 cagtctcgaa cttaagctgc agtgactctc ttaaggtagc cttgcagaag ttggtcgtga
      361 ggcactgggc aggtaagtat caaggttaca agacaggttt aaggagacca atagaaactg
      421 ggcttgtcga gacagagaag actcttgcgt ttctgatagg cacctattgg tcttactgac
      481 atccactttg cctttctctc cacaggtgtc cactcccagt tcaattacag ctcttaaggc
      541 tagagtactt aatacgactc actataggct agccaccatg gaagacgcca aaaacataaa
      601 gaaaggcccg gcgccattct atccgctgga agatggaacc gctggagagc aactgcataa
      661 ggctatgaag agatacgccc tggttcctgg aacaattgct tttacagatg cacatatcga
      721 ggtggacatc acttacgctg agtacttcga aatgtccgtt cggttggcag aagctatgaa
      781 acgatatggg ctgaatacaa atcacagaat cgtcgtatgc agtgaaaact ctcttcaatt
      841 ctttatgccg gtgttgggcg cgttatttat cggagttgca gttgcgcccg cgaacgacat
      901 ttataatgaa cgtgaattgc tcaacagtat gggcatttcg cagcctaccg tggtgttcgt
      961 ttccaaaaag gggttgcaaa aaattttgaa cgtgcaaaaa aagctcccaa tcatccaaaa
     1021 aattattatc atggattcta aaacggatta ccagggattt cagtcgatgt acacgttcgt
     1081 cacatctcat ctacctcccg gttttaatga atacgatttt gtgccagagt ccttcgatag
     1141 ggacaagaca attgcactga tcatgaactc ctctggatct actggtctgc ctaaaggtgt
     1201 cgctctgcct catagaactg cctgcgtgag attctcgcat gccagagatc ctatttttgg
     1261 caatcaaatc attccggata ctgcgatttt aagtgttgtt ccattccatc acggttttgg
     1321 aatgtttact acactcggat atttgatatg tggatttcga gtcgtcttaa tgtatagatt
     1381 tgaagaagag ctgtttctga ggagccttca ggattacaag attcaaagtg cgctgctggt
     1441 gccaacccta ttctccttct tcgccaaaag cactctgatt gacaaatacg atttatctaa
     1501 tttacacgaa attgcttctg gtggcgctcc cctctctaag gaagtcgggg aagcggttgc
     1561 caagaggttc catctgccag gtatcaggca aggatatggg ctcactgaga ctacatcagc
     1621 tattctgatt acacccgagg gggatgataa accgggcgcg gtcggtaaag ttgttccatt
     1681 ttttgaagcg aaggttgtgg atctggatac cgggaaaacg ctgggcgtta atcaaagagg
     1741 cgaactgtgt gtgagaggtc ctatgattat gtccggttat gtaaacaatc cggaagcgac
     1801 caacgccttg attgacaagg atggatggct acattctgga gacatagctt actgggacga
     1861 agacgaacac ttcttcatcg ttgaccgcct gaagtctctg attaagtaca aaggctatca
     1921 ggtggctccc gctgaattgg aatccatctt gctccaacac cccaacatct tcgacgcagg
     1981 tgtcgcaggt cttcccgacg atgacgccgg tgaacttccc gccgccgttg ttgttttgga
     2041 gcacggaaag acgatgacgg aaaaagagat cgtggattac gtcgccagtc aagtaacaac
     2101 cgcgaaaaag ttgcgcggag gagttgtgtt tgtggacgaa gtaccgaaag gtcttaccgg
     2161 aaaactcgac gcaagaaaaa tcagagagat cctcataaag gccaagaagg gcggaaagat
     2221 cgccgtggaa ttcatggacg agaaggtgtt caccaaggag ctggaccagt ggatcgagca
     2281 gctgaacgag tgcaagcagc tgtccgagtc ccaggtcaag agcctctgcg agaaggctaa
     2341 agaaatcctg acaaaagaat ccaacgtgca agaggttcga tgtccagtta ctgtctgtgg
     2401 agatgtgcat gggcaatttc atgatctcat ggaactgttt agaattggtg gcaaatcacc
     2461 agatacaaat tacttgttta tgggagatta tgttgacaga ggatattatt cagttgaaac
     2521 agttacactg cttgtagctc ttaaggttcg ttaccgtgaa cgcatcacca ttcttcgagg
     2581 gaatcatgag agcagacaga tcacacaagt ttatggtttc tatgatgaat gtttaagaaa
     2641 atatggaaat gcaaatgttt ggaaatattt tacagatctt tttgactatc ttcctctcac
     2701 tgccttggtg gatgggcaga tcttctgtct acatggtggt ctctcgccat ctatagatac
     2761 actggatcat atcagagcac ttgatcgcct acaagaagtt ccccatgagg gtccaatgtg
     2821 tgacttgctg tggtcagatc cagatgaccg tggtggttgg ggtatatctc ctcgaggagc
     2881 tggttacacc tttgggcaag atatttctga gacatttaat catgccaatg gcctcacgtt
     2941 ggtgtctaga gctcaccagc tagtgatgga gggatataac tggtgccagg accggaatgt
     3001 agtaacgatt ttcagtgctc caaactattg ttatcgttgt ggtaaccaag ctgcaatcat
     3061 ggaacttgac gatactctaa aatactcttt cttgcagttt gacccagcac ctcgtagagg
     3121 cgagccacat gttactcgtc gtaccccaga ctacttcctg taatgtcgac ccgggcggcc
     3181 gcttcccttt agtgagggtt aatgcttcga gcagacatga taagatacat tgatgagttt
     3241 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     3301 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     3361 cattttatgt ttcaggttca gggggagatg tgggaggttt tttaaagcaa gtaaaacctc
     3421 tacaaatgtg gtaaaatccg ataaggatcg atccgggctg gcgtaatagc gaagaggccc
     3481 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggacg cgccctgtag
     3541 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3601 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3661 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3721 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3781 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3841 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     3901 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     3961 caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt
     4021 atttcacacc gcatacgcgg atctgcgcag caccatggcc tgaaataacc tctgaaagag
     4081 gaacttggtt aggtaccttc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     4141 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4201 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4261 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4321 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4381 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4441 gcctaggctt ttgcaaaaag cttgattctt ctgacacaac agtctcgaac ttaaggctag
     4501 agccaccatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     4561 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     4621 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     4681 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     4741 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     4801 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     4861 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     4921 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     4981 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     5041 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     5101 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     5161 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     5221 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     5281 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     5341 cccaacctgc catcacgatg gccgcaataa aatatcttta ttttcattac atctgtgtgt
     5401 tggttttttg tgtgaatcga tagcgataag gatccgcgta tggtgcactc tcagtacaat
     5461 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     5521 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     5581 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     5641 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     5701 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     5761 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     5821 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     5881 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     5941 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     6001 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     6061 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     6121 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     6181 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     6241 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     6301 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     6361 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     6421 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     6481 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     6541 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     6601 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     6661 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     6721 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     6781 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     6841 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6901 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6961 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     7021 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     7081 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     7141 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     7201 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     7261 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     7321 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     7381 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     7441 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     7501 catggctcga cagatcttca atattggcca ttagccatat tattcattgg ttatatagca
     7561 taaatcaata ttggctattg gccattgcat acgttgtatc tatatcataa tatgtacatt
     7621 tatattggct catgtccaat atgaccgcca tgttggcatt gattattgac tagttattaa
     7681 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     7741 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     7801 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     7861 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtccgccc
     7921 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     7981 cgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catg
//
