LOCUS       sequence-471827-        6028 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express ERE response element in luciferase reporter assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6028)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6028)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pGL4 [luc2P/ERE/Hygro] Vector
FEATURES             Location/Qualifiers
     source          1..6028
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        95..126
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      153..162
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             159..1808
                     /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             1812..1931
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    1983..2104
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1994..2013)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2048..2067
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2298..2655
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2298..2489
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2321..2341)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2506..2641
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2568..2587
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2686..3723
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             3702..3710
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    3747..3795
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(3951..3968)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4122..4710)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4202..4221)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4910..5701,5702..5770))
                     /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    5875..5923
                     /note="synthetic polyadenylation signal"
     misc_feature    5937..6028
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5977..5996
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtaccg tcaggtcaca gtgacctgat gtcaggtcac agtgacctga
       61 tgtcaggtca cagtgacctg ataagcttag acactagagg gtatataatg gaagctcgac
      121 ttccagcttg gcaatccggt actgttggta aagccaccat ggaagatgcc aaaaacatta
      181 agaagggccc agcgccattc tacccactcg aagacgggac cgccggcgag cagctgcaca
      241 aagccatgaa gcgctacgcc ctggtgcccg gcaccatcgc ctttaccgac gcacatatcg
      301 aggtggacat tacctacgcc gagtacttcg agatgagcgt tcggctggca gaagctatga
      361 agcgctatgg gctgaataca aaccatcgga tcgtggtgtg cagcgagaat agcttgcagt
      421 tcttcatgcc cgtgttgggt gccctgttca tcggtgtggc tgtggcccca gctaacgaca
      481 tctacaacga gcgcgagctg ctgaacagca tgggcatcag ccagcccacc gtcgtattcg
      541 tgagcaagaa agggctgcaa aagatcctca acgtgcaaaa gaagctaccg atcatacaaa
      601 agatcatcat catggatagc aagaccgact accagggctt ccaaagcatg tacaccttcg
      661 tgacttccca tttgccaccc ggcttcaacg agtacgactt cgtgcccgag agcttcgacc
      721 gggacaaaac catcgccctg atcatgaaca gtagtggcag taccggattg cccaagggcg
      781 tagccctacc gcaccgcacc gcttgtgtcc gattcagtca tgcccgcgac cccatcttcg
      841 gcaaccagat catccccgac accgctatcc tcagcgtggt gccatttcac cacggcttcg
      901 gcatgttcac cacgctgggc tacttgatct gcggctttcg ggtcgtgctc atgtaccgct
      961 tcgaggagga gctattcttg cgcagcttgc aagactataa gattcaatct gccctgctgg
     1021 tgcccacact atttagcttc ttcgctaaga gcactctcat cgacaagtac gacctaagca
     1081 acttgcacga gatcgccagc ggcggggcgc cgctcagcaa ggaggtaggt gaggccgtgg
     1141 ccaaacgctt ccacctacca ggcatccgcc agggctacgg cctgacagaa acaaccagcg
     1201 ccattctgat cacccccgaa ggggacgaca agcctggcgc agtaggcaag gtggtgccct
     1261 tcttcgaggc taaggtggtg gacttggaca ccggtaagac actgggtgtg aaccagcgcg
     1321 gcgagctgtg cgtccgtggc cccatgatca tgagcggcta cgttaacaac cccgaggcta
     1381 caaacgctct catcgacaag gacggctggc tgcacagcgg cgacatcgcc tactgggacg
     1441 aggacgagca cttcttcatc gtggaccggc tgaagagcct gatcaaatac aagggctacc
     1501 aggtagcccc agccgaactg gagagcatcc tgctgcaaca ccccaacatc ttcgacgccg
     1561 gggtcgccgg cctgcccgac gacgatgccg gcgagctgcc cgccgcagtc gtcgtgctgg
     1621 aacacggtaa aaccatgacc gagaaggaga tcgtggacta tgtggccagc caggttacaa
     1681 ccgccaagaa gctgcgcggt ggtgttgtgt tcgtggacga ggtgcctaaa ggactgaccg
     1741 gcaagttgga cgcccgcaag atccgcgaga ttctcattaa ggccaagaag ggcggcaaga
     1801 tcgccgtgaa ttctcacggc ttccctcccg aggtggagga gcaggccgcc ggcaccctgc
     1861 ccatgagctg cgcccaggag agcggcatgg atagacaccc tgctgcttgc gccagcgcca
     1921 ggatcaacgt ctaaggccgc gactctagag tcggggcggc cggccgcttc gagcagacat
     1981 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     2041 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     2101 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     2161 tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc gataaggatc cgtttgcgta
     2221 ttgggcgctc ttccgctgat ctgcgcagca ccatggcctg aaataacctc tgaaagagga
     2281 acttggttag ctaccttctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     2341 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     2401 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     2461 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     2521 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     2581 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     2641 ctaggctttt gcaaaaagct cgattcttct gacactagcg ccaccatgaa gaagcccgaa
     2701 ctcaccgcta ccagcgttga aaaatttctc atcgagaagt tcgacagtgt gagcgacctg
     2761 atgcagttgt cggagggcga agagagccga gccttcagct tcgatgtcgg cggacgcggc
     2821 tatgtactgc gggtgaatag ctgcgctgat ggcttctaca aagaccgcta cgtgtaccgc
     2881 cacttcgcca gcgctgcact acccatcccc gaagtgttgg acatcggcga gttcagcgag
     2941 agcctgacat actgcatcag tagacgcgcc caaggcgtta ctctccaaga cctccccgaa
     3001 acagagctgc ctgctgtgtt acagcctgtc gccgaagcta tggatgctat tgccgccgcc
     3061 gacctcagtc aaaccagcgg cttcggccca ttcgggcccc aaggcatcgg ccagtacaca
     3121 acctggcggg atttcatttg cgccattgct gatccccatg tctaccactg gcagaccgtg
     3181 atggacgaca ccgtgtccgc cagcgtagct caagccctgg acgaactgat gctgtgggcc
     3241 gaagactgtc ccgaggtgcg ccacctcgtc catgccgact tcggcagcaa caacgtcctg
     3301 accgacaacg gccgcatcac cgccgtaatc gactggtccg aagctatgtt cggggacagt
     3361 cagtacgagg tggccaacat cttcttctgg cggccctggc tggcttgcat ggagcagcag
     3421 actcgctact tcgagcgccg gcatcccgag ctggccggca gccctcgtct gcgagcctac
     3481 atgctgcgca tcggcctgga tcagctctac cagagcctcg tggacggcaa cttcgacgat
     3541 gctgcctggg ctcaaggccg ctgcgatgcc atcgtccgca gcggggccgg caccgtcggt
     3601 cgcacacaaa tcgctcgccg gagcgcagcc gtatggaccg acggctgcgt cgaggtgctg
     3661 gccgacagcg gcaaccgccg gcccagtaca cgaccgcgcg ctaaggaggt aggtcgagtt
     3721 taaactctag aaccggtcat ggccgcaata aaatatcttt attttcatta catctgtgtg
     3781 ttggtttttt gtgtgttcga actagatgct gtcgaccgat gcccttgaga gccttcaacc
     3841 cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct
     3901 tctttatcat gcaactcgta ggacaggtgc cggcagcgct cttccgcttc ctcgctcact
     3961 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4021 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4081 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4141 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4201 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4261 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4321 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4381 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4441 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4501 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4561 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4621 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4681 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4741 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4801 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4861 gagtaaactt ggtctgacag cggccgcaaa tgctaaacca ctgcagtggt taccagtgct
     4921 tgatcagtga ggcaccgatc tcagcgatct gcctatttcg ttcgtccata gtggcctgac
     4981 tccccgtcgt gtagatcact acgattcgtg agggcttacc atcaggcccc agcgcagcaa
     5041 tgatgccgcg agagccgcgt tcaccggccc ccgatttgtc agcaatgaac cagccagcag
     5101 ggagggccga gcgaagaagt ggtcctgcta ctttgtccgc ctccatccag tctatgagct
     5161 gctgtcgtga tgctagagta agaagttcgc cagtgagtag tttccgaaga gttgtggcca
     5221 ttgctactgg catcgtggta tcacgctcgt cgttcggtat ggcttcgttc aactctggtt
     5281 cccagcggtc aagccgggtc acatgatcac ccatattatg aagaaatgca gtcagctcct
     5341 tagggcctcc gatcgttgtc agaagtaagt tggccgcggt gttgtcgctc atggtaatgg
     5401 cagcactaca caattctctt accgtcatgc catccgtaag atgcttttcc gtgaccggcg
     5461 agtactcaac caagtcgttt tgtgagtagt gtatacggcg accaagctgc tcttgcccgg
     5521 cgtctatacg ggacaacacc gcgccacata gcagtacttt gaaagtgctc atcatcggga
     5581 atcgttcttc ggggcggaaa gactcaagga tcttgccgct attgagatcc agttcgatat
     5641 agcccactct tgcacccagt tgatcttcag catcttttac tttcaccagc gtttcggggt
     5701 gtgcaaaaac aggcaagcaa aatgccgcaa agaagggaat gagtgcgaca cgaaaatgtt
     5761 ggatgctcat actcgtcctt tttcaatatt attgaagcat ttatcagggt tactagtacg
     5821 tctctcaagg ataagtaagt aatattaagg tacgggaggt attggacagg ccgcaataaa
     5881 atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat agtactaaca
     5941 tacgctctcc atcaaaacaa aacgaaacaa aacaaactag caaaataggc tgtccccagt
     6001 gcaagtgcag gtgccagaac atttctct
//
