LOCUS       sequence-209376-        6544 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CMV luciferase reporter - firefly luciferase driven by CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6544)
  AUTHORS   Grohar PJ, Woldemichael GM, Griffin LB, Mendoza A, Chen QR, Yeung C,
            Currier DG, Davis S, Khanna C, Khan J, McMahon JB, Helman LJ
  TITLE     Identification of an inhibitor of the EWS-FLI1 oncogenic 
            transcription factor by high-throughput screening.
  JOURNAL   J Natl Cancer Inst. 2011 Jun 22;103(12):962-78. doi: 
            10.1093/jnci/djr156. Epub 2011 Jun 8.
  PUBMED    21653923
REFERENCE   2  (bases 1 to 6544)
  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: 
            "10.1093/jnci/djr156"; journalName: "J Natl Cancer Inst"; date: 
            "2011-06-22- 22"; volume: "103"; issue: "12"; pages: "962-78"
COMMENT     Sequence Label: pGL4.18 CMV-Luc
FEATURES             Location/Qualifiers
     source          1..6544
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      729..738
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             735..2384
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     CDS             2388..2507
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    2559..2680
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2570..2589)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2624..2643
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2874..3231
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2874..3065
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2897..2917)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3082..3217
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3144..3163
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3262..4056
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4081..4129
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4285..4302)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4456..5044)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4536..4555)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5244..6035,6036..6104))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    6209..6257
                     /note="synthetic polyadenylation signal"
     misc_feature    6271..6362
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6311..6330
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcactag aagctttatt gcggtagttt atcacagtta aattgctaac gcagtcagtg
      661 ggctagcctc gaggatatca agatctggcc tcggcggcca agcttggcaa tccggtactg
      721 ttggtaaagc caccatggaa gatgccaaaa acattaagaa gggcccagcg ccattctacc
      781 cactcgaaga cgggaccgcc ggcgagcagc tgcacaaagc catgaagcgc tacgccctgg
      841 tgcccggcac catcgccttt accgacgcac atatcgaggt ggacattacc tacgccgagt
      901 acttcgagat gagcgttcgg ctggcagaag ctatgaagcg ctatgggctg aatacaaacc
      961 atcggatcgt ggtgtgcagc gagaatagct tgcagttctt catgcccgtg ttgggtgccc
     1021 tgttcatcgg tgtggctgtg gccccagcta acgacatcta caacgagcgc gagctgctga
     1081 acagcatggg catcagccag cccaccgtcg tattcgtgag caagaaaggg ctgcaaaaga
     1141 tcctcaacgt gcaaaagaag ctaccgatca tacaaaagat catcatcatg gatagcaaga
     1201 ccgactacca gggcttccaa agcatgtaca ccttcgtgac ttcccatttg ccacccggct
     1261 tcaacgagta cgacttcgtg cccgagagct tcgaccggga caaaaccatc gccctgatca
     1321 tgaacagtag tggcagtacc ggattgccca agggcgtagc cctaccgcac cgcaccgctt
     1381 gtgtccgatt cagtcatgcc cgcgacccca tcttcggcaa ccagatcatc cccgacaccg
     1441 ctatcctcag cgtggtgcca tttcaccacg gcttcggcat gttcaccacg ctgggctact
     1501 tgatctgcgg ctttcgggtc gtgctcatgt accgcttcga ggaggagcta ttcttgcgca
     1561 gcttgcaaga ctataagatt caatctgccc tgctggtgcc cacactattt agcttcttcg
     1621 ctaagagcac tctcatcgac aagtacgacc taagcaactt gcacgagatc gccagcggcg
     1681 gggcgccgct cagcaaggag gtaggtgagg ccgtggccaa acgcttccac ctaccaggca
     1741 tccgccaggg ctacggcctg acagaaacaa ccagcgccat tctgatcacc cccgaagggg
     1801 acgacaagcc tggcgcagta ggcaaggtgg tgcccttctt cgaggctaag gtggtggact
     1861 tggacaccgg taagacactg ggtgtgaacc agcgcggcga gctgtgcgtc cgtggcccca
     1921 tgatcatgag cggctacgtt aacaaccccg aggctacaaa cgctctcatc gacaaggacg
     1981 gctggctgca cagcggcgac atcgcctact gggacgagga cgagcacttc ttcatcgtgg
     2041 accggctgaa gagcctgatc aaatacaagg gctaccaggt agccccagcc gaactggaga
     2101 gcatcctgct gcaacacccc aacatcttcg acgccggggt cgccggcctg cccgacgacg
     2161 atgccggcga gctgcccgcc gcagtcgtcg tgctggaaca cggtaaaacc atgaccgaga
     2221 aggagatcgt ggactatgtg gccagccagg ttacaaccgc caagaagctg cgcggtggtg
     2281 ttgtgttcgt ggacgaggtg cctaaaggac tgaccggcaa gttggacgcc cgcaagatcc
     2341 gcgagattct cattaaggcc aagaagggcg gcaagatcgc cgtgaattct cacggcttcc
     2401 ctcccgaggt ggaggagcag gccgccggca ccctgcccat gagctgcgcc caggagagcg
     2461 gcatggatag acaccctgct gcttgcgcca gcgccaggat caacgtctaa ggccgcgact
     2521 ctagagtcgg ggcggccggc cgcttcgagc agacatgata agatacattg atgagtttgg
     2581 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     2641 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     2701 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     2761 caaatgtggt aaaatcgata aggatccgtt tgcgtattgg gcgctcttcc gctgatctgc
     2821 gcagcaccat ggcctgaaat aacctctgaa agaggaactt ggttagctac cttctgaggc
     2881 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     2941 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3001 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3061 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3121 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     3181 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcgat
     3241 tcttctgaca ctagcgccac catgatcgaa caagacggcc tccatgctgg cagtcccgca
     3301 gcttgggtcg aacgcttgtt cgggtacgac tgggcccagc agaccatcgg atgtagcgat
     3361 gcggccgtgt tccgtctaag cgctcaaggc cggcccgtgc tgttcgtgaa gaccgacctg
     3421 agcggcgccc tgaacgagct tcaagacgag gctgcccgcc tgagctggct ggccaccacc
     3481 ggcgtaccct gcgccgctgt gttggatgtt gtgaccgaag ccggccggga ctggctgctg
     3541 ctgggcgagg tccctggcca ggatctgctg agcagccacc ttgcccccgc tgagaaggtt
     3601 tctatcatgg ccgatgcaat gcggcgcctg cacaccctgg accccgctac ctgccccttc
     3661 gaccaccagg ctaagcatcg gatcgagcgt gctcggaccc gcatggaggc cggcctggtg
     3721 gaccaggacg acctggacga ggagcatcag ggcctggccc ccgctgaact gttcgcccga
     3781 ctgaaagccc gcatgccgga cggtgaggac ctggttgtca cacacggaga tgcctgcctc
     3841 cctaacatca tggtcgagaa tggccgcttc tccggcttca tcgactgcgg tcgcctagga
     3901 gttgccgacc gctaccagga catcgccctg gccacccgcg acatcgctga ggagcttggc
     3961 ggcgagtggg ccgaccgctt cttagtcttg tacggcatcg cagctcccga cagccagcgc
     4021 atcgccttct accgcttgct cgacgagttc ttttaatgat ctagaaccgg tcatggccgc
     4081 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtgt tcgaactaga
     4141 tgctgtcgac cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg
     4201 ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag
     4261 gtgccggcag cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4321 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4381 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4441 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4501 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4561 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4621 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4681 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4741 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4801 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4861 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4921 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4981 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5041 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5101 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5161 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagcggccg
     5221 caaatgctaa accactgcag tggttaccag tgcttgatca gtgaggcacc gatctcagcg
     5281 atctgcctat ttcgttcgtc catagtggcc tgactccccg tcgtgtagat cactacgatt
     5341 cgtgagggct taccatcagg ccccagcgca gcaatgatgc cgcgagagcc gcgttcaccg
     5401 gcccccgatt tgtcagcaat gaaccagcca gcagggaggg ccgagcgaag aagtggtcct
     5461 gctactttgt ccgcctccat ccagtctatg agctgctgtc gtgatgctag agtaagaagt
     5521 tcgccagtga gtagtttccg aagagttgtg gccattgcta ctggcatcgt ggtatcacgc
     5581 tcgtcgttcg gtatggcttc gttcaactct ggttcccagc ggtcaagccg ggtcacatga
     5641 tcacccatat tatgaagaaa tgcagtcagc tccttagggc ctccgatcgt tgtcagaagt
     5701 aagttggccg cggtgttgtc gctcatggta atggcagcac tacacaattc tcttaccgtc
     5761 atgccatccg taagatgctt ttccgtgacc ggcgagtact caaccaagtc gttttgtgag
     5821 tagtgtatac ggcgaccaag ctgctcttgc ccggcgtcta tacgggacaa caccgcgcca
     5881 catagcagta ctttgaaagt gctcatcatc gggaatcgtt cttcggggcg gaaagactca
     5941 aggatcttgc cgctattgag atccagttcg atatagccca ctcttgcacc cagttgatct
     6001 tcagcatctt ttactttcac cagcgtttcg gggtgtgcaa aaacaggcaa gcaaaatgcc
     6061 gcaaagaagg gaatgagtgc gacacgaaaa tgttggatgc tcatactcgt cctttttcaa
     6121 tattattgaa gcatttatca gggttactag tacgtctctc aaggataagt aagtaatatt
     6181 aaggtacggg aggtattgga caggccgcaa taaaatatct ttattttcat tacatctgtg
     6241 tgttggtttt ttgtgtgaat cgatagtact aacatacgct ctccatcaaa acaaaacgaa
     6301 acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca gaacatttct
     6361 ctggcctaac tggccggtac ctgagctcgc tagcggccta actggcctca atattggcca
     6421 ttagccatat tattcattgg ttatatagca taaatcaata ttggctattg gccattgcat
     6481 acgttgtatc tatatcataa tatgtacatt tatattggct catgtccaat atgaccgcca
     6541 tgtt
//
