LOCUS       sequence-260666-        7697 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NR0B1 luciferase reporter - firefly luciferase driven by NR0B1 
            promoter .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7697)
  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 7697)
  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 NR0B1-Luc
FEATURES             Location/Qualifiers
     source          1..7697
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    511..602
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(543..562)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    616..664
                     /note="synthetic polyadenylation signal"
     CDS             join(769..837,838..1629)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1829..2417
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2318..2337
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2571..2588
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    2744..2792
                     /note="synthetic polyadenylation signal"
     CDS             complement(2817..3611)
                     /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"
     promoter        complement(3642..3999)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3656..3791
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3710..3729)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        3808..3999
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     3956..3976
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    4193..4314
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4230..4249)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4284..4303
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(4366..4485)
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     CDS             complement(4489..6138)
                     /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"
     regulatory      6135..6144
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ccttccttcc tttccttcct tccttccttc cttccttcct tccttccttc ccttccttcc
       61 ttccttcctc cctccctccc tccctccctc cctccttctc tcactccttc tctctctctc
      121 tttctttctc tttttcttct tctccaggac ctggaattct cagcataaga ggaaaaccta
      181 ggagaccccc attttgcttg cagcgttaaa tttcattctg cctgtgagag gccctaaata
      241 gcacagtggt taaatccctg gaactagcac tagattgcct cagttttaat cctggccctg
      301 ccactttgta gcatcataac cttgggcaat gtgttagttt ctttgtcgct gctttactta
      361 ccacaaatgt agtggcttaa gtcaacacaa acaacacgaa ttttttatgt tataattctg
      421 gaagccagaa gactgaaatc agtttcactg agctaaagtc aaggtgacat ttcttctgga
      481 agttctagga cggtaccggc cagttaggcc agagaaatgt tctggcacct gcacttgcac
      541 tggggacagc ctattttgct agtttgtttt gtttcgtttt gttttgatgg agagcgtatg
      601 ttagtactat cgattcacac aaaaaaccaa cacacagatg taatgaaaat aaagatattt
      661 tattgcggcc tgtccaatac ctcccgtacc ttaatattac ttacttatcc ttgagagacg
      721 tactagtaac cctgataaat gcttcaataa tattgaaaaa ggacgagtat gagcatccaa
      781 cattttcgtg tcgcactcat tcccttcttt gcggcatttt gcttgcctgt ttttgcacac
      841 cccgaaacgc tggtgaaagt aaaagatgct gaagatcaac tgggtgcaag agtgggctat
      901 atcgaactgg atctcaatag cggcaagatc cttgagtctt tccgccccga agaacgattc
      961 ccgatgatga gcactttcaa agtactgcta tgtggcgcgg tgttgtcccg tatagacgcc
     1021 gggcaagagc agcttggtcg ccgtatacac tactcacaaa acgacttggt tgagtactcg
     1081 ccggtcacgg aaaagcatct tacggatggc atgacggtaa gagaattgtg tagtgctgcc
     1141 attaccatga gcgacaacac cgcggccaac ttacttctga caacgatcgg aggccctaag
     1201 gagctgactg catttcttca taatatgggt gatcatgtga cccggcttga ccgctgggaa
     1261 ccagagttga acgaagccat accgaacgac gagcgtgata ccacgatgcc agtagcaatg
     1321 gccacaactc ttcggaaact actcactggc gaacttctta ctctagcatc acgacagcag
     1381 ctcatagact ggatggaggc ggacaaagta gcaggaccac ttcttcgctc ggccctccct
     1441 gctggctggt tcattgctga caaatcgggg gccggtgaac gcggctctcg cggcatcatt
     1501 gctgcgctgg ggcctgatgg taagccctca cgaatcgtag tgatctacac gacggggagt
     1561 caggccacta tggacgaacg aaataggcag atcgctgaga tcggtgcctc actgatcaag
     1621 cactggtaac cactgcagtg gtttagcatt tgcggccgct gtcagaccaa gtttactcat
     1681 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     1741 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     1801 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     1861 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     1921 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     1981 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     2041 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     2101 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     2161 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     2221 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     2281 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     2341 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     2401 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     2461 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     2521 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     2581 tgagcgagga agcggaagag cgctgccggc acctgtccta cgagttgcat gataaagaag
     2641 acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg
     2701 ttgaaggctc tcaagggcat cggtcgacag catctagttc gaacacacaa aaaaccaaca
     2761 cacagatgta atgaaaataa agatatttta ttgcggccat gaccggttct agatcattaa
     2821 aagaactcgt cgagcaagcg gtagaaggcg atgcgctggc tgtcgggagc tgcgatgccg
     2881 tacaagacta agaagcggtc ggcccactcg ccgccaagct cctcagcgat gtcgcgggtg
     2941 gccagggcga tgtcctggta gcggtcggca actcctaggc gaccgcagtc gatgaagccg
     3001 gagaagcggc cattctcgac catgatgtta gggaggcagg catctccgtg tgtgacaacc
     3061 aggtcctcac cgtccggcat gcgggctttc agtcgggcga acagttcagc gggggccagg
     3121 ccctgatgct cctcgtccag gtcgtcctgg tccaccaggc cggcctccat gcgggtccga
     3181 gcacgctcga tccgatgctt agcctggtgg tcgaaggggc aggtagcggg gtccagggtg
     3241 tgcaggcgcc gcattgcatc ggccatgata gaaaccttct cagcgggggc aaggtggctg
     3301 ctcagcagat cctggccagg gacctcgccc agcagcagcc agtcccggcc ggcttcggtc
     3361 acaacatcca acacagcggc gcagggtacg ccggtggtgg ccagccagct caggcgggca
     3421 gcctcgtctt gaagctcgtt cagggcgccg ctcaggtcgg tcttcacgaa cagcacgggc
     3481 cggccttgag cgcttagacg gaacacggcc gcatcgctac atccgatggt ctgctgggcc
     3541 cagtcgtacc cgaacaagcg ttcgacccaa gctgcgggac tgccagcatg gaggccgtct
     3601 tgttcgatca tggtggcgct agtgtcagaa gaatcgagct ttttgcaaaa gcctaggcct
     3661 ccaaaaaagc ctcctcacta cttctggaat agctcagagg cagaggcggc ctcggcctct
     3721 gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact gggcggagtt
     3781 aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt gagatgcatg
     3841 ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt tgctgactaa
     3901 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacaccct
     3961 aactgacaca cattccacag ctggttcttt ccgcctcaga aggtagctaa ccaagttcct
     4021 ctttcagagg ttatttcagg ccatggtgct gcgcagatca gcggaagagc gcccaatacg
     4081 caaacggatc cttatcgatt ttaccacatt tgtagaggtt ttacttgctt taaaaaacct
     4141 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     4201 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     4261 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     4321 ctgctcgaag cggccggccg ccccgactct agagtcgcgg ccttagacgt tgatcctggc
     4381 gctggcgcaa gcagcagggt gtctatccat gccgctctcc tgggcgcagc tcatgggcag
     4441 ggtgccggcg gcctgctcct ccacctcggg agggaagccg tgagaattca cggcgatctt
     4501 gccgcccttc ttggccttaa tgagaatctc gcggatcttg cgggcgtcca acttgccggt
     4561 cagtccttta ggcacctcgt ccacgaacac aacaccaccg cgcagcttct tggcggttgt
     4621 aacctggctg gccacatagt ccacgatctc cttctcggtc atggttttac cgtgttccag
     4681 cacgacgact gcggcgggca gctcgccggc atcgtcgtcg ggcaggccgg cgaccccggc
     4741 gtcgaagatg ttggggtgtt gcagcaggat gctctccagt tcggctgggg ctacctggta
     4801 gcccttgtat ttgatcaggc tcttcagccg gtccacgatg aagaagtgct cgtcctcgtc
     4861 ccagtaggcg atgtcgccgc tgtgcagcca gccgtccttg tcgatgagag cgtttgtagc
     4921 ctcggggttg ttaacgtagc cgctcatgat catggggcca cggacgcaca gctcgccgcg
     4981 ctggttcaca cccagtgtct taccggtgtc caagtccacc accttagcct cgaagaaggg
     5041 caccaccttg cctactgcgc caggcttgtc gtccccttcg ggggtgatca gaatggcgct
     5101 ggttgtttct gtcaggccgt agccctggcg gatgcctggt aggtggaagc gtttggccac
     5161 ggcctcacct acctccttgc tgagcggcgc cccgccgctg gcgatctcgt gcaagttgct
     5221 taggtcgtac ttgtcgatga gagtgctctt agcgaagaag ctaaatagtg tgggcaccag
     5281 cagggcagat tgaatcttat agtcttgcaa gctgcgcaag aatagctcct cctcgaagcg
     5341 gtacatgagc acgacccgaa agccgcagat caagtagccc agcgtggtga acatgccgaa
     5401 gccgtggtga aatggcacca cgctgaggat agcggtgtcg gggatgatct ggttgccgaa
     5461 gatggggtcg cgggcatgac tgaatcggac acaagcggtg cggtgcggta gggctacgcc
     5521 cttgggcaat ccggtactgc cactactgtt catgatcagg gcgatggttt tgtcccggtc
     5581 gaagctctcg ggcacgaagt cgtactcgtt gaagccgggt ggcaaatggg aagtcacgaa
     5641 ggtgtacatg ctttggaagc cctggtagtc ggtcttgcta tccatgatga tgatcttttg
     5701 tatgatcggt agcttctttt gcacgttgag gatcttttgc agccctttct tgctcacgaa
     5761 tacgacggtg ggctggctga tgcccatgct gttcagcagc tcgcgctcgt tgtagatgtc
     5821 gttagctggg gccacagcca caccgatgaa cagggcaccc aacacgggca tgaagaactg
     5881 caagctattc tcgctgcaca ccacgatccg atggtttgta ttcagcccat agcgcttcat
     5941 agcttctgcc agccgaacgc tcatctcgaa gtactcggcg taggtaatgt ccacctcgat
     6001 atgtgcgtcg gtaaaggcga tggtgccggg caccagggcg tagcgcttca tggctttgtg
     6061 cagctgctcg ccggcggtcc cgtcttcgag tgggtagaat ggcgctgggc ccttcttaat
     6121 gtttttggca tcttccatgg tggctttacc aacagtaccg gattgccaag cttggccgcc
     6181 gaggccagat cttgatatcc tcgaggctag ccttcgcgct cataagcatg ttgtagagga
     6241 tgctgccctg ccactggtgg ttctcgcccg ccatggcccg cggcgcccgt agcccagtcc
     6301 tgcccagtgg ctgcctcctg ggacctattt atacctagcg cgcacgcagg cgcgtcggcc
     6361 gcctcagtct gggcaagggc gcggaggcgg gcggtgtgtg cgggggggcg cggtgctgcg
     6421 ctccggtgtg ttcgcccatg acctcggtgg ttcgaggggg cgggtgctct taaaagctgg
     6481 aaatggaaga acagcagcgt gggagctcct ggacactttc ctccttcctc tcattacccc
     6541 tttccctgtc tcaggaagac cggaggagag cgagcgcctg gatgctgttg caactctttc
     6601 cattgaaccc tcgcatcaaa tttgttccct ttattgggag ggcaggggaa aagaggaaac
     6661 atctacttct atgctctttg taaactttca ttaaattatc acagtttgtg tgacttatat
     6721 ttgtatatgg atagggaggt ctcattttct gagaccttga aaaatacaga acatagacaa
     6781 gcatcatatg ggaactcggt ggaaatgccc aatctcaggc tccaaccaga ctttctaact
     6841 ctgaatctgc attttaacgg tgtccccagg gattcacatg cacctagacg tttgaaaagc
     6901 actgctctaa atatgctatc ataagcgctc agtttaattc catcagtatt tataagcgcc
     6961 cactatgtcc tggatgctgt gttaagagag ctgggggcag cgagcaggat gtaaaagtgc
     7021 ctaaaactta ggaatataat aattacgagg ggataatatt gttacagtga gaattgtttt
     7081 cgttttagta tatacttttt tttcttttaa cttttaggtt caggagtata tgtgcgggtt
     7141 tgttatataa gtgaattgct tgtcgtgggg gtttggtgta cagattattt catcacccag
     7201 gtaatagtat ccaataggta ttttttctca ttctctccct cttcccaccc tccaccttca
     7261 agtaggttcc agtgtctgtt gttcccctcc tagtgtccag gtgatctctt tatttagctc
     7321 ccacttgcaa gtgagaacat gtgatatttg gttttctgtt cctgcattac tttgctaagc
     7381 atgatggccg gcagctctca ttctttttca tggccgcagt gagaaatttt gatccttgta
     7441 aaagtattcc actggtaagt gtgtaattag tatttctttt cccacccttc ctaagtgaat
     7501 aatgataaat agaagcatca ggaagcctgg atccattagt cttcacttgc cacacagctg
     7561 tgaagacatg gggcagggaa gggtgttggt atataccagc tgatacagaa tcatttttac
     7621 tttttccctc cctcccatcc tccctctttc tttttttgct gtttcttttc cttccttcct
     7681 tccttccttc cttcctt
//
