LOCUS       sequence-252752-        8095 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Vector for LUMIER assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8095)
  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 8095)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-PP1A
FEATURES             Location/Qualifiers
     source          1..8095
                     /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    3282..3403
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3293..3312)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3347..3366
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3589..4044
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3676..3695)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3886..3907
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4161..4518
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4161..4352
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4184..4204)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4369..4504
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4569..5363
                     /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(4623..4642)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5233..5252
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5427..5475
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5516..5535)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5635..5657
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5695..5713)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5781..5885
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5886..5954,5955..6746)
                     /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(6104..6123)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6917..7505
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7406..7425
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7716..8095
                     /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 ttcatgtccg acagcgagaa gctcaacctg gactcgatca tcgggcgcct
     2281 gctggaagtg cagggctcgc ggcctggcaa gaatgtacag ctgacagaga acgagatccg
     2341 cggtctgtgc ctgaaatctc gggagatttt tctgagccag cccattcttc tggagctgga
     2401 ggcacccctc aagatctgcg gtgacataca cggccagtac tacgaccttc tgcgactatt
     2461 tgagtatggc ggtttccctc ccgagagcaa ctacctcttt ctgggggact atgtggacag
     2521 gggcaagcag tccttggaga ccatctgcct gctgctggcc tataagatca agtaccccga
     2581 gaacttcttc ctgctccgtg ggaaccacga gtgtgccagc atcaaccgca tctatggttt
     2641 ctacgatgag tgcaagagac gctacaacat caaactgtgg aaaaccttca ctgactgctt
     2701 caactgcctg cccatcgcgg ccatagtgga cgaaaagatc ttctgctgcc acggaggcct
     2761 gtccccggac ctgcagtcta tggagcagat tcggcggatc atgcggccca cagatgtgcc
     2821 tgaccagggc ctgctgtgtg acctgctgtg gtctgaccct gacaaggacg tgcagggctg
     2881 gggcgagaac gaccgtggcg tctcttttgc ctttggagcc gaggtggtgg ccaagttcct
     2941 ccacaagcac gacttggacc tcatctgccg agcacaccag gtggtagaag acggctacga
     3001 gttctttgcc aagcggcagc tggtgacact tttctcagct cccaactact gtggcgagtt
     3061 tgacaatgct ggcgccatga tgagtgtgga cgagaccctc atgtgctctt tccagatcct
     3121 caagcccgcc gacaagaaca aggggaagta cgggcagttc agtggcctga accctggagg
     3181 ccgacccatc accccacccc gcaattccgc caaagccaag aaatagtcga cccgggcggc
     3241 cgcttccctt tagtgagggt taatgcttcg agcagacatg ataagataca ttgatgagtt
     3301 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     3361 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     3421 tcattttatg tttcaggttc agggggagat gtgggaggtt ttttaaagca agtaaaacct
     3481 ctacaaatgt ggtaaaatcc gataaggatc gatccgggct ggcgtaatag cgaagaggcc
     3541 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggac gcgccctgta
     3601 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3661 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3721 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3781 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     3841 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3901 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     3961 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     4021 acaaaatatt aacgcttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg
     4081 tatttcacac cgcatacgcg gatctgcgca gcaccatggc ctgaaataac ctctgaaaga
     4141 ggaacttggt taggtacctt ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     4201 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4261 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4321 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4381 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4441 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4501 ggcctaggct tttgcaaaaa gcttgattct tctgacacaa cagtctcgaa cttaaggcta
     4561 gagccaccat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     4621 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4681 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4741 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4801 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4861 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4921 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4981 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     5041 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     5101 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     5161 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     5221 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     5281 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     5341 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     5401 gcccaacctg ccatcacgat ggccgcaata aaatatcttt attttcatta catctgtgtg
     5461 ttggtttttt gtgtgaatcg atagcgataa ggatccgcgt atggtgcact ctcagtacaa
     5521 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     5581 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     5641 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     5701 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     5761 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5821 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5881 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5941 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     6001 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     6061 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     6121 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     6181 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     6241 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     6301 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     6361 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     6421 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     6481 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     6541 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     6601 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     6661 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     6721 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     6781 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6841 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6901 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6961 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     7021 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     7081 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     7141 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     7201 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     7261 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     7321 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     7381 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     7441 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     7501 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     7561 acatggctcg acagatcttc aatattggcc attagccata ttattcattg gttatatagc
     7621 ataaatcaat attggctatt ggccattgca tacgttgtat ctatatcata atatgtacat
     7681 ttatattggc tcatgtccaa tatgaccgcc atgttggcat tgattattga ctagttatta
     7741 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     7801 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     7861 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     7921 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtccgcc
     7981 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     8041 acgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatg
//
