LOCUS       sequence-194148-        6096 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase reporter plasmid containing three tandem repeats of the 
            ER stress response element-II (ERSE-II).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6096)
  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 6096)
  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-ERSE2-luc2P-Hygro
FEATURES             Location/Qualifiers
     source          1..6096
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        163..194
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      221..230
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             227..1876
                     /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             1880..1999
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    2051..2172
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2062..2081)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2116..2135
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2366..2723
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2366..2557
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2389..2409)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2574..2709
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2636..2655
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2754..3791
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             3770..3778
                     /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    3815..3863
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4019..4036)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4190..4778)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4270..4289)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4978..5769,5770..5838))
                     /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    5943..5991
                     /note="synthetic polyadenylation signal"
     misc_feature    6005..6096
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6045..6064
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcgcta gcggacgccg attgggccac gttgggagag
       61 tgcctggacg ccgattgggc cacgttggga gagtgcctgg acgccgattg ggccacgttg
      121 ggagagtgcc tagatctggc ctcggcggcc aagcttagac actagagggt atataatgga
      181 agctcgactt ccagcttggc aatccggtac tgttggtaaa gccaccatgg aagatgccaa
      241 aaacattaag aagggcccag cgccattcta cccactcgaa gacgggaccg ccggcgagca
      301 gctgcacaaa gccatgaagc gctacgccct ggtgcccggc accatcgcct ttaccgacgc
      361 acatatcgag gtggacatta cctacgccga gtacttcgag atgagcgttc ggctggcaga
      421 agctatgaag cgctatgggc tgaatacaaa ccatcggatc gtggtgtgca gcgagaatag
      481 cttgcagttc ttcatgcccg tgttgggtgc cctgttcatc ggtgtggctg tggccccagc
      541 taacgacatc tacaacgagc gcgagctgct gaacagcatg ggcatcagcc agcccaccgt
      601 cgtattcgtg agcaagaaag ggctgcaaaa gatcctcaac gtgcaaaaga agctaccgat
      661 catacaaaag atcatcatca tggatagcaa gaccgactac cagggcttcc aaagcatgta
      721 caccttcgtg acttcccatt tgccacccgg cttcaacgag tacgacttcg tgcccgagag
      781 cttcgaccgg gacaaaacca tcgccctgat catgaacagt agtggcagta ccggattgcc
      841 caagggcgta gccctaccgc accgcaccgc ttgtgtccga ttcagtcatg cccgcgaccc
      901 catcttcggc aaccagatca tccccgacac cgctatcctc agcgtggtgc catttcacca
      961 cggcttcggc atgttcacca cgctgggcta cttgatctgc ggctttcggg tcgtgctcat
     1021 gtaccgcttc gaggaggagc tattcttgcg cagcttgcaa gactataaga ttcaatctgc
     1081 cctgctggtg cccacactat ttagcttctt cgctaagagc actctcatcg acaagtacga
     1141 cctaagcaac ttgcacgaga tcgccagcgg cggggcgccg ctcagcaagg aggtaggtga
     1201 ggccgtggcc aaacgcttcc acctaccagg catccgccag ggctacggcc tgacagaaac
     1261 aaccagcgcc attctgatca cccccgaagg ggacgacaag cctggcgcag taggcaaggt
     1321 ggtgcccttc ttcgaggcta aggtggtgga cttggacacc ggtaagacac tgggtgtgaa
     1381 ccagcgcggc gagctgtgcg tccgtggccc catgatcatg agcggctacg ttaacaaccc
     1441 cgaggctaca aacgctctca tcgacaagga cggctggctg cacagcggcg acatcgccta
     1501 ctgggacgag gacgagcact tcttcatcgt ggaccggctg aagagcctga tcaaatacaa
     1561 gggctaccag gtagccccag ccgaactgga gagcatcctg ctgcaacacc ccaacatctt
     1621 cgacgccggg gtcgccggcc tgcccgacga cgatgccggc gagctgcccg ccgcagtcgt
     1681 cgtgctggaa cacggtaaaa ccatgaccga gaaggagatc gtggactatg tggccagcca
     1741 ggttacaacc gccaagaagc tgcgcggtgg tgttgtgttc gtggacgagg tgcctaaagg
     1801 actgaccggc aagttggacg cccgcaagat ccgcgagatt ctcattaagg ccaagaaggg
     1861 cggcaagatc gccgtgaatt ctcacggctt ccctcccgag gtggaggagc aggccgccgg
     1921 caccctgccc atgagctgcg cccaggagag cggcatggat agacaccctg ctgcttgcgc
     1981 cagcgccagg atcaacgtct aaggccgcga ctctagagtc ggggcggccg gccgcttcga
     2041 gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     2101 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     2161 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     2221 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaggatccg
     2281 tttgcgtatt gggcgctctt ccgctgatct gcgcagcacc atggcctgaa ataacctctg
     2341 aaagaggaac ttggttagct accttctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     2401 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2461 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2521 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     2581 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     2641 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     2701 ttggaggcct aggcttttgc aaaaagctcg attcttctga cactagcgcc accatgaaga
     2761 agcccgaact caccgctacc agcgttgaaa aatttctcat cgagaagttc gacagtgtga
     2821 gcgacctgat gcagttgtcg gagggcgaag agagccgagc cttcagcttc gatgtcggcg
     2881 gacgcggcta tgtactgcgg gtgaatagct gcgctgatgg cttctacaaa gaccgctacg
     2941 tgtaccgcca cttcgccagc gctgcactac ccatccccga agtgttggac atcggcgagt
     3001 tcagcgagag cctgacatac tgcatcagta gacgcgccca aggcgttact ctccaagacc
     3061 tccccgaaac agagctgcct gctgtgttac agcctgtcgc cgaagctatg gatgctattg
     3121 ccgccgccga cctcagtcaa accagcggct tcggcccatt cgggccccaa ggcatcggcc
     3181 agtacacaac ctggcgggat ttcatttgcg ccattgctga tccccatgtc taccactggc
     3241 agaccgtgat ggacgacacc gtgtccgcca gcgtagctca agccctggac gaactgatgc
     3301 tgtgggccga agactgtccc gaggtgcgcc acctcgtcca tgccgacttc ggcagcaaca
     3361 acgtcctgac cgacaacggc cgcatcaccg ccgtaatcga ctggtccgaa gctatgttcg
     3421 gggacagtca gtacgaggtg gccaacatct tcttctggcg gccctggctg gcttgcatgg
     3481 agcagcagac tcgctacttc gagcgccggc atcccgagct ggccggcagc cctcgtctgc
     3541 gagcctacat gctgcgcatc ggcctggatc agctctacca gagcctcgtg gacggcaact
     3601 tcgacgatgc tgcctgggct caaggccgct gcgatgccat cgtccgcagc ggggccggca
     3661 ccgtcggtcg cacacaaatc gctcgccgga gcgcagccgt atggaccgac ggctgcgtcg
     3721 aggtgctggc cgacagcggc aaccgccggc ccagtacacg accgcgcgct aaggaggtag
     3781 gtcgagttta aactctagaa ccggtcatgg ccgcaataaa atatctttat tttcattaca
     3841 tctgtgtgtt ggttttttgt gtgttcgaac tagatgctgt cgaccgatgc ccttgagagc
     3901 cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat
     3961 gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct tccgcttcct
     4021 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4081 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4141 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4201 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4261 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4321 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4381 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4441 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4501 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4561 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4621 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4681 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4741 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4801 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4861 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4921 gtatatatga gtaaacttgg tctgacagcg gccgcaaatg ctaaaccact gcagtggtta
     4981 ccagtgcttg atcagtgagg caccgatctc agcgatctgc ctatttcgtt cgtccatagt
     5041 ggcctgactc cccgtcgtgt agatcactac gattcgtgag ggcttaccat caggccccag
     5101 cgcagcaatg atgccgcgag agccgcgttc accggccccc gatttgtcag caatgaacca
     5161 gccagcaggg agggccgagc gaagaagtgg tcctgctact ttgtccgcct ccatccagtc
     5221 tatgagctgc tgtcgtgatg ctagagtaag aagttcgcca gtgagtagtt tccgaagagt
     5281 tgtggccatt gctactggca tcgtggtatc acgctcgtcg ttcggtatgg cttcgttcaa
     5341 ctctggttcc cagcggtcaa gccgggtcac atgatcaccc atattatgaa gaaatgcagt
     5401 cagctcctta gggcctccga tcgttgtcag aagtaagttg gccgcggtgt tgtcgctcat
     5461 ggtaatggca gcactacaca attctcttac cgtcatgcca tccgtaagat gcttttccgt
     5521 gaccggcgag tactcaacca agtcgttttg tgagtagtgt atacggcgac caagctgctc
     5581 ttgcccggcg tctatacggg acaacaccgc gccacatagc agtactttga aagtgctcat
     5641 catcgggaat cgttcttcgg ggcggaaaga ctcaaggatc ttgccgctat tgagatccag
     5701 ttcgatatag cccactcttg cacccagttg atcttcagca tcttttactt tcaccagcgt
     5761 ttcggggtgt gcaaaaacag gcaagcaaaa tgccgcaaag aagggaatga gtgcgacacg
     5821 aaaatgttgg atgctcatac tcgtcctttt tcaatattat tgaagcattt atcagggtta
     5881 ctagtacgtc tctcaaggat aagtaagtaa tattaaggta cgggaggtat tggacaggcc
     5941 gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag
     6001 tactaacata cgctctccat caaaacaaaa cgaaacaaaa caaactagca aaataggctg
     6061 tccccagtgc aagtgcaggt gccagaacat ttctct
//
