LOCUS       sequence-194146-        6084 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luferase reporter plasmid containing three tandem repeats of the ER 
            stress response element-I (ERSE-I).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6084)
  AUTHORS   Fujita H, Bando T, Oyadomari S, Ochiai K, Watanabe M, Kumon H, 
            Ohuchi H
  TITLE     Dkk3/REIC, an N-glycosylated Protein, Is a Physiological Endoplasmic
            Reticulum Stress Inducer in the Mouse Adrenal Gland.
  JOURNAL   Acta Med Okayama. 2020 Jun;74(3):199-208. doi: 10.18926/AMO/59950.
  PUBMED    32577017
REFERENCE   2  (bases 1 to 6084)
  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"; journalName: 
            "Acta Med Okayama"; date: "2020-06"; volume: "74"; issue: "3"; 
            pages: "199-208"
COMMENT     Sequence Label: pGL4-ERSE1-luc2P-Hygro
FEATURES             Location/Qualifiers
     source          1..6084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        151..182
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      209..218
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             215..1864
                     /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             1868..1987
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    2039..2160
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2050..2069)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2104..2123
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2354..2711
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2354..2545
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2377..2397)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2562..2697
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2624..2643
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2742..3779
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             3758..3766
                     /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    3803..3851
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4007..4024)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4178..4766)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4258..4277)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4966..5757,5758..5826))
                     /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    5931..5979
                     /note="synthetic polyadenylation signal"
     misc_feature    5993..6084
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6033..6052
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcgcta gcccttcacc aatcggcggc ctccacgacg
       61 gccttcacca atcggcggcc tccacgacgg ccttcaccaa tcggcggcct ccacgacgga
      121 gatctggcct cggcggccaa gcttagacac tagagggtat ataatggaag ctcgacttcc
      181 agcttggcaa tccggtactg ttggtaaagc caccatggaa gatgccaaaa acattaagaa
      241 gggcccagcg ccattctacc cactcgaaga cgggaccgcc ggcgagcagc tgcacaaagc
      301 catgaagcgc tacgccctgg tgcccggcac catcgccttt accgacgcac atatcgaggt
      361 ggacattacc tacgccgagt acttcgagat gagcgttcgg ctggcagaag ctatgaagcg
      421 ctatgggctg aatacaaacc atcggatcgt ggtgtgcagc gagaatagct tgcagttctt
      481 catgcccgtg ttgggtgccc tgttcatcgg tgtggctgtg gccccagcta acgacatcta
      541 caacgagcgc gagctgctga acagcatggg catcagccag cccaccgtcg tattcgtgag
      601 caagaaaggg ctgcaaaaga tcctcaacgt gcaaaagaag ctaccgatca tacaaaagat
      661 catcatcatg gatagcaaga ccgactacca gggcttccaa agcatgtaca ccttcgtgac
      721 ttcccatttg ccacccggct tcaacgagta cgacttcgtg cccgagagct tcgaccggga
      781 caaaaccatc gccctgatca tgaacagtag tggcagtacc ggattgccca agggcgtagc
      841 cctaccgcac cgcaccgctt gtgtccgatt cagtcatgcc cgcgacccca tcttcggcaa
      901 ccagatcatc cccgacaccg ctatcctcag cgtggtgcca tttcaccacg gcttcggcat
      961 gttcaccacg ctgggctact tgatctgcgg ctttcgggtc gtgctcatgt accgcttcga
     1021 ggaggagcta ttcttgcgca gcttgcaaga ctataagatt caatctgccc tgctggtgcc
     1081 cacactattt agcttcttcg ctaagagcac tctcatcgac aagtacgacc taagcaactt
     1141 gcacgagatc gccagcggcg gggcgccgct cagcaaggag gtaggtgagg ccgtggccaa
     1201 acgcttccac ctaccaggca tccgccaggg ctacggcctg acagaaacaa ccagcgccat
     1261 tctgatcacc cccgaagggg acgacaagcc tggcgcagta ggcaaggtgg tgcccttctt
     1321 cgaggctaag gtggtggact tggacaccgg taagacactg ggtgtgaacc agcgcggcga
     1381 gctgtgcgtc cgtggcccca tgatcatgag cggctacgtt aacaaccccg aggctacaaa
     1441 cgctctcatc gacaaggacg gctggctgca cagcggcgac atcgcctact gggacgagga
     1501 cgagcacttc ttcatcgtgg accggctgaa gagcctgatc aaatacaagg gctaccaggt
     1561 agccccagcc gaactggaga gcatcctgct gcaacacccc aacatcttcg acgccggggt
     1621 cgccggcctg cccgacgacg atgccggcga gctgcccgcc gcagtcgtcg tgctggaaca
     1681 cggtaaaacc atgaccgaga aggagatcgt ggactatgtg gccagccagg ttacaaccgc
     1741 caagaagctg cgcggtggtg ttgtgttcgt ggacgaggtg cctaaaggac tgaccggcaa
     1801 gttggacgcc cgcaagatcc gcgagattct cattaaggcc aagaagggcg gcaagatcgc
     1861 cgtgaattct cacggcttcc ctcccgaggt ggaggagcag gccgccggca ccctgcccat
     1921 gagctgcgcc caggagagcg gcatggatag acaccctgct gcttgcgcca gcgccaggat
     1981 caacgtctaa ggccgcgact ctagagtcgg ggcggccggc cgcttcgagc agacatgata
     2041 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     2101 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     2161 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     2221 taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatccgtt tgcgtattgg
     2281 gcgctcttcc gctgatctgc gcagcaccat ggcctgaaat aacctctgaa agaggaactt
     2341 ggttagctac cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     2401 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2461 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2521 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     2581 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     2641 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     2701 gcttttgcaa aaagctcgat tcttctgaca ctagcgccac catgaagaag cccgaactca
     2761 ccgctaccag cgttgaaaaa tttctcatcg agaagttcga cagtgtgagc gacctgatgc
     2821 agttgtcgga gggcgaagag agccgagcct tcagcttcga tgtcggcgga cgcggctatg
     2881 tactgcgggt gaatagctgc gctgatggct tctacaaaga ccgctacgtg taccgccact
     2941 tcgccagcgc tgcactaccc atccccgaag tgttggacat cggcgagttc agcgagagcc
     3001 tgacatactg catcagtaga cgcgcccaag gcgttactct ccaagacctc cccgaaacag
     3061 agctgcctgc tgtgttacag cctgtcgccg aagctatgga tgctattgcc gccgccgacc
     3121 tcagtcaaac cagcggcttc ggcccattcg ggccccaagg catcggccag tacacaacct
     3181 ggcgggattt catttgcgcc attgctgatc cccatgtcta ccactggcag accgtgatgg
     3241 acgacaccgt gtccgccagc gtagctcaag ccctggacga actgatgctg tgggccgaag
     3301 actgtcccga ggtgcgccac ctcgtccatg ccgacttcgg cagcaacaac gtcctgaccg
     3361 acaacggccg catcaccgcc gtaatcgact ggtccgaagc tatgttcggg gacagtcagt
     3421 acgaggtggc caacatcttc ttctggcggc cctggctggc ttgcatggag cagcagactc
     3481 gctacttcga gcgccggcat cccgagctgg ccggcagccc tcgtctgcga gcctacatgc
     3541 tgcgcatcgg cctggatcag ctctaccaga gcctcgtgga cggcaacttc gacgatgctg
     3601 cctgggctca aggccgctgc gatgccatcg tccgcagcgg ggccggcacc gtcggtcgca
     3661 cacaaatcgc tcgccggagc gcagccgtat ggaccgacgg ctgcgtcgag gtgctggccg
     3721 acagcggcaa ccgccggccc agtacacgac cgcgcgctaa ggaggtaggt cgagtttaaa
     3781 ctctagaacc ggtcatggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg
     3841 ttttttgtgt gttcgaacta gatgctgtcg accgatgccc ttgagagcct tcaacccagt
     3901 cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt
     3961 tatcatgcaa ctcgtaggac aggtgccggc agcgctcttc cgcttcctcg ctcactgact
     4021 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4081 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4141 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4201 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4261 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4321 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4381 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4441 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4501 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4561 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     4621 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4681 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4741 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4801 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4861 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     4921 aaacttggtc tgacagcggc cgcaaatgct aaaccactgc agtggttacc agtgcttgat
     4981 cagtgaggca ccgatctcag cgatctgcct atttcgttcg tccatagtgg cctgactccc
     5041 cgtcgtgtag atcactacga ttcgtgaggg cttaccatca ggccccagcg cagcaatgat
     5101 gccgcgagag ccgcgttcac cggcccccga tttgtcagca atgaaccagc cagcagggag
     5161 ggccgagcga agaagtggtc ctgctacttt gtccgcctcc atccagtcta tgagctgctg
     5221 tcgtgatgct agagtaagaa gttcgccagt gagtagtttc cgaagagttg tggccattgc
     5281 tactggcatc gtggtatcac gctcgtcgtt cggtatggct tcgttcaact ctggttccca
     5341 gcggtcaagc cgggtcacat gatcacccat attatgaaga aatgcagtca gctccttagg
     5401 gcctccgatc gttgtcagaa gtaagttggc cgcggtgttg tcgctcatgg taatggcagc
     5461 actacacaat tctcttaccg tcatgccatc cgtaagatgc ttttccgtga ccggcgagta
     5521 ctcaaccaag tcgttttgtg agtagtgtat acggcgacca agctgctctt gcccggcgtc
     5581 tatacgggac aacaccgcgc cacatagcag tactttgaaa gtgctcatca tcgggaatcg
     5641 ttcttcgggg cggaaagact caaggatctt gccgctattg agatccagtt cgatatagcc
     5701 cactcttgca cccagttgat cttcagcatc ttttactttc accagcgttt cggggtgtgc
     5761 aaaaacaggc aagcaaaatg ccgcaaagaa gggaatgagt gcgacacgaa aatgttggat
     5821 gctcatactc gtcctttttc aatattattg aagcatttat cagggttact agtacgtctc
     5881 tcaaggataa gtaagtaata ttaaggtacg ggaggtattg gacaggccgc aataaaatat
     5941 ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagta ctaacatacg
     6001 ctctccatca aaacaaaacg aaacaaaaca aactagcaaa ataggctgtc cccagtgcaa
     6061 gtgcaggtgc cagaacattt ctct
//
