LOCUS       sequence-252700-        8445 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 8445)
  AUTHORS   Maruyama J, Inami K, Michishita F, Jiang X, Iwasa H, Nakagawa K, 
            Ishigami-Yuasa M, Kagechika H, Miyamura N, Hirayama J, Nishina H, 
            Nogawa D, Yamamoto K, Hata Y
  TITLE     Novel YAP1 Activator, Identified by Transcription-Based Functional 
            Screen, Limits Multiple Myeloma Growth.
  JOURNAL   Mol Cancer Res. 2018 Feb;16(2):197-211. doi: 
            10.1158/1541-7786.MCR-17-0382. Epub 2017 Oct 23.
  PUBMED    29061667
REFERENCE   2  (bases 1 to 8445)
  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"; doi: 
            "10.1158/1541-7786.MCR-17-0382"; journalName: "Mol Cancer Res"; 
            date: "2018-02"; volume: "16"; issue: "2"; pages: "197-211"
COMMENT     Sequence Label: pCIneoLuc-TEAD4
FEATURES             Location/Qualifiers
     source          1..8445
                     /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    3632..3753
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3643..3662)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3697..3716
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3939..4394
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4026..4045)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4236..4257
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4511..4868
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4511..4702
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4534..4554)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4719..4854
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4919..5713
                     /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(4973..4992)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5583..5602
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5777..5825
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5866..5885)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5985..6007
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6045..6063)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6131..6235
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6236..6304,6305..7096)
                     /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(6454..6473)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7267..7855
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7756..7775
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        8066..8445
                     /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 ttcgagggca cggccggcac cattacctcc aacgagtgga gctctcccac
     2281 ctcccctgag gggagcaccg cctctggggg cagtcaggca ctggacaagc ccatcgacaa
     2341 tgacgcagag ggcgtgtgga gcccggatat tgagcagagt ttccaggagg ccctcgccat
     2401 ctacccgccc tgtggcaggc gcaaaatcat cctgtcggac gagggtaaga tgtatggtcg
     2461 gaacgagctg attgcccgct acatcaagct ccggacaggg aagacccgca ccaggaagca
     2521 ggtctccagc cacatccagg tgctggctcg tcgcaaagct cgcgagatcc aggccaagct
     2581 aaaggaccag gcagctaagg acaaggccct gcagagcatg gctgccatgt cgtctgcaca
     2641 gatcatctcc gccacggcct tccacagtag catggccctc gcccggggcc ccggccgccc
     2701 agcagtctca gggttttggc aaggagcttt gccaggccaa gccggaacgt cccatgatgt
     2761 gaagcctttc tctcagcaaa cctatgctgt ccagcctccg ctgcctctgc cagggtttga
     2821 gtctcctgca gggcccgccc catcgccctc tgcgcccccg gcacccccat ggcagggccg
     2881 cagcgtggcc agctccaagc tctggatgtt ggagttctct gccttcctgg agcagcagca
     2941 ggacccggac acgtacaaca agcacctgtt cgtgcacatt ggccagtcca gcccaagcta
     3001 cagcgacccc tacctcgaag ccgtggacat ccgccaaatc tatgacaaat tcccggagaa
     3061 aaagggtgga ctcaaggatc tcttcgaacg gggaccctcc aatgcctttt ttcttgtgaa
     3121 gttctgggca gacctcaaca ccaacatcga ggatgaaggc ggctccttct atggggtctc
     3181 cagccagtat gagagccccg agaacatgat catcacctgc tccacgaagg tctgctcttt
     3241 cggcaagcag gtggtggaga aagttgagac agagtatgct cgctatgaga atggacacta
     3301 ctcttaccgc atccaccggt ccccgctctg tgagtacatg atcaacttca tccacaagct
     3361 caagcacctc cctgagaagt acatgatgaa cagcgtgctg gagaacttca ccatcctgca
     3421 ggtggtcacc aacagagaca cacaggagac cttgctgtgc attgcctatg tctttgaggt
     3481 gtcagccagt gagcacgggg ctcagcacca catctacagg ctggtgaaag aatgagtcga
     3541 caattgtgga tccgctctag agtcgacctg caggcatgca agcttgcggc cgcttccctt
     3601 tagtgagggt taatgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc
     3661 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     3721 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     3781 tttcaggttc agggggagat gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
     3841 ggtaaaatcc gataaggatc gatccgggct ggcgtaatag cgaagaggcc cgcaccgatc
     3901 gcccttccca acagttgcgc agcctgaatg gcgaatggac gcgccctgta gcggcgcatt
     3961 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     4021 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4081 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4141 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4201 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4261 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     4321 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     4381 aacgcttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     4441 cgcatacgcg gatctgcgca gcaccatggc ctgaaataac ctctgaaaga ggaacttggt
     4501 taggtacctt ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     4561 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4621 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4681 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4741 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4801 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4861 tttgcaaaaa gcttgattct tctgacacaa cagtctcgaa cttaaggcta gagccaccat
     4921 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     4981 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     5041 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     5101 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     5161 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     5221 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     5281 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     5341 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     5401 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     5461 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     5521 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     5581 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     5641 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     5701 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     5761 ccatcacgat ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt
     5821 gtgtgaatcg atagcgataa ggatccgcgt atggtgcact ctcagtacaa tctgctctga
     5881 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     5941 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     6001 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     6061 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     6121 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     6181 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     6241 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     6301 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     6361 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     6421 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     6481 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     6541 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     6601 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     6661 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     6721 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     6781 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     6841 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     6901 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     6961 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     7021 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     7081 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     7141 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     7201 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     7261 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     7321 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     7381 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     7441 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     7501 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     7561 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     7621 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     7681 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     7741 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     7801 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     7861 cagtaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatggctcg
     7921 acagatcttc aatattggcc attagccata ttattcattg gttatatagc ataaatcaat
     7981 attggctatt ggccattgca tacgttgtat ctatatcata atatgtacat ttatattggc
     8041 tcatgtccaa tatgaccgcc atgttggcat tgattattga ctagttatta atagtaatca
     8101 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
     8161 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     8221 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     8281 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtccgcc ccctattgac
     8341 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt acgggacttt
     8401 cctacttggc agtacatcta cgtattagtc atcgctatta ccatg
//
