LOCUS       sequence-194135-        6054 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luferase reporter plasmid containing three tandem repeats of the 
            amino acid response element (AARE).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6054)
  AUTHORS   Miyake M, Nomura A, Ogura A, Takehana K, Kitahara Y, Takahara K, 
            Tsugawa K, Miyamoto C, Miura N, Sato R, Kurahashi K, Harding HP, 
            Oyadomari M, Ron D, Oyadomari S
  TITLE     Skeletal muscle-specific eukaryotic translation initiation factor 
            2alpha phosphorylation controls amino acid metabolism and fibroblast
            growth factor 21-mediated non-cell-autonomous energy metabolism.
  JOURNAL   FASEB J. 2016 Feb;30(2):798-812. doi: 10.1096/fj.15-275990. Epub 
            2015 Oct 20.
  PUBMED    26487695
REFERENCE   2  (bases 1 to 6054)
  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: 
            "10.1096/fj.15-275990"; journalName: "FASEB J"; date: "2016-02"; 
            volume: "30"; issue: "2"; pages: "798-812"
COMMENT     Sequence Label: pGL4-AARE-luc2P-Hygro
FEATURES             Location/Qualifiers
     source          1..6054
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        121..152
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      179..188
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             185..1834
                     /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             1838..1957
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    2009..2130
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2020..2039)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2074..2093
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2324..2681
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2324..2515
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2347..2367)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2532..2667
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2594..2613
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2712..3749
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             3728..3736
                     /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    3773..3821
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(3977..3994)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4148..4736)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4228..4247)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4936..5727,5728..5796))
                     /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    5901..5949
                     /note="synthetic polyadenylation signal"
     misc_feature    5963..6054
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6003..6022
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcgcta gcaacattgc atcatccccg caacattgca
       61 tcatccccgc aacattgcat catccccgca gatctggcct cggcggccaa gcttagacac
      121 tagagggtat ataatggaag ctcgacttcc agcttggcaa tccggtactg ttggtaaagc
      181 caccatggaa gatgccaaaa acattaagaa gggcccagcg ccattctacc cactcgaaga
      241 cgggaccgcc ggcgagcagc tgcacaaagc catgaagcgc tacgccctgg tgcccggcac
      301 catcgccttt accgacgcac atatcgaggt ggacattacc tacgccgagt acttcgagat
      361 gagcgttcgg ctggcagaag ctatgaagcg ctatgggctg aatacaaacc atcggatcgt
      421 ggtgtgcagc gagaatagct tgcagttctt catgcccgtg ttgggtgccc tgttcatcgg
      481 tgtggctgtg gccccagcta acgacatcta caacgagcgc gagctgctga acagcatggg
      541 catcagccag cccaccgtcg tattcgtgag caagaaaggg ctgcaaaaga tcctcaacgt
      601 gcaaaagaag ctaccgatca tacaaaagat catcatcatg gatagcaaga ccgactacca
      661 gggcttccaa agcatgtaca ccttcgtgac ttcccatttg ccacccggct tcaacgagta
      721 cgacttcgtg cccgagagct tcgaccggga caaaaccatc gccctgatca tgaacagtag
      781 tggcagtacc ggattgccca agggcgtagc cctaccgcac cgcaccgctt gtgtccgatt
      841 cagtcatgcc cgcgacccca tcttcggcaa ccagatcatc cccgacaccg ctatcctcag
      901 cgtggtgcca tttcaccacg gcttcggcat gttcaccacg ctgggctact tgatctgcgg
      961 ctttcgggtc gtgctcatgt accgcttcga ggaggagcta ttcttgcgca gcttgcaaga
     1021 ctataagatt caatctgccc tgctggtgcc cacactattt agcttcttcg ctaagagcac
     1081 tctcatcgac aagtacgacc taagcaactt gcacgagatc gccagcggcg gggcgccgct
     1141 cagcaaggag gtaggtgagg ccgtggccaa acgcttccac ctaccaggca tccgccaggg
     1201 ctacggcctg acagaaacaa ccagcgccat tctgatcacc cccgaagggg acgacaagcc
     1261 tggcgcagta ggcaaggtgg tgcccttctt cgaggctaag gtggtggact tggacaccgg
     1321 taagacactg ggtgtgaacc agcgcggcga gctgtgcgtc cgtggcccca tgatcatgag
     1381 cggctacgtt aacaaccccg aggctacaaa cgctctcatc gacaaggacg gctggctgca
     1441 cagcggcgac atcgcctact gggacgagga cgagcacttc ttcatcgtgg accggctgaa
     1501 gagcctgatc aaatacaagg gctaccaggt agccccagcc gaactggaga gcatcctgct
     1561 gcaacacccc aacatcttcg acgccggggt cgccggcctg cccgacgacg atgccggcga
     1621 gctgcccgcc gcagtcgtcg tgctggaaca cggtaaaacc atgaccgaga aggagatcgt
     1681 ggactatgtg gccagccagg ttacaaccgc caagaagctg cgcggtggtg ttgtgttcgt
     1741 ggacgaggtg cctaaaggac tgaccggcaa gttggacgcc cgcaagatcc gcgagattct
     1801 cattaaggcc aagaagggcg gcaagatcgc cgtgaattct cacggcttcc ctcccgaggt
     1861 ggaggagcag gccgccggca ccctgcccat gagctgcgcc caggagagcg gcatggatag
     1921 acaccctgct gcttgcgcca gcgccaggat caacgtctaa ggccgcgact ctagagtcgg
     1981 ggcggccggc cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca
     2041 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     2101 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     2161 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     2221 aaaatcgata aggatccgtt tgcgtattgg gcgctcttcc gctgatctgc gcagcaccat
     2281 ggcctgaaat aacctctgaa agaggaactt ggttagctac cttctgaggc ggaaagaacc
     2341 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2401 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     2461 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     2521 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     2581 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     2641 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcgat tcttctgaca
     2701 ctagcgccac catgaagaag cccgaactca ccgctaccag cgttgaaaaa tttctcatcg
     2761 agaagttcga cagtgtgagc gacctgatgc agttgtcgga gggcgaagag agccgagcct
     2821 tcagcttcga tgtcggcgga cgcggctatg tactgcgggt gaatagctgc gctgatggct
     2881 tctacaaaga ccgctacgtg taccgccact tcgccagcgc tgcactaccc atccccgaag
     2941 tgttggacat cggcgagttc agcgagagcc tgacatactg catcagtaga cgcgcccaag
     3001 gcgttactct ccaagacctc cccgaaacag agctgcctgc tgtgttacag cctgtcgccg
     3061 aagctatgga tgctattgcc gccgccgacc tcagtcaaac cagcggcttc ggcccattcg
     3121 ggccccaagg catcggccag tacacaacct ggcgggattt catttgcgcc attgctgatc
     3181 cccatgtcta ccactggcag accgtgatgg acgacaccgt gtccgccagc gtagctcaag
     3241 ccctggacga actgatgctg tgggccgaag actgtcccga ggtgcgccac ctcgtccatg
     3301 ccgacttcgg cagcaacaac gtcctgaccg acaacggccg catcaccgcc gtaatcgact
     3361 ggtccgaagc tatgttcggg gacagtcagt acgaggtggc caacatcttc ttctggcggc
     3421 cctggctggc ttgcatggag cagcagactc gctacttcga gcgccggcat cccgagctgg
     3481 ccggcagccc tcgtctgcga gcctacatgc tgcgcatcgg cctggatcag ctctaccaga
     3541 gcctcgtgga cggcaacttc gacgatgctg cctgggctca aggccgctgc gatgccatcg
     3601 tccgcagcgg ggccggcacc gtcggtcgca cacaaatcgc tcgccggagc gcagccgtat
     3661 ggaccgacgg ctgcgtcgag gtgctggccg acagcggcaa ccgccggccc agtacacgac
     3721 cgcgcgctaa ggaggtaggt cgagtttaaa ctctagaacc ggtcatggcc gcaataaaat
     3781 atctttattt tcattacatc tgtgtgttgg ttttttgtgt gttcgaacta gatgctgtcg
     3841 accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac
     3901 tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc
     3961 agcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4021 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4081 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4141 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4201 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4261 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4321 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4381 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4441 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4501 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4561 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4621 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4681 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4741 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4801 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     4861 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagcggc cgcaaatgct
     4921 aaaccactgc agtggttacc agtgcttgat cagtgaggca ccgatctcag cgatctgcct
     4981 atttcgttcg tccatagtgg cctgactccc cgtcgtgtag atcactacga ttcgtgaggg
     5041 cttaccatca ggccccagcg cagcaatgat gccgcgagag ccgcgttcac cggcccccga
     5101 tttgtcagca atgaaccagc cagcagggag ggccgagcga agaagtggtc ctgctacttt
     5161 gtccgcctcc atccagtcta tgagctgctg tcgtgatgct agagtaagaa gttcgccagt
     5221 gagtagtttc cgaagagttg tggccattgc tactggcatc gtggtatcac gctcgtcgtt
     5281 cggtatggct tcgttcaact ctggttccca gcggtcaagc cgggtcacat gatcacccat
     5341 attatgaaga aatgcagtca gctccttagg gcctccgatc gttgtcagaa gtaagttggc
     5401 cgcggtgttg tcgctcatgg taatggcagc actacacaat tctcttaccg tcatgccatc
     5461 cgtaagatgc ttttccgtga ccggcgagta ctcaaccaag tcgttttgtg agtagtgtat
     5521 acggcgacca agctgctctt gcccggcgtc tatacgggac aacaccgcgc cacatagcag
     5581 tactttgaaa gtgctcatca tcgggaatcg ttcttcgggg cggaaagact caaggatctt
     5641 gccgctattg agatccagtt cgatatagcc cactcttgca cccagttgat cttcagcatc
     5701 ttttactttc accagcgttt cggggtgtgc aaaaacaggc aagcaaaatg ccgcaaagaa
     5761 gggaatgagt gcgacacgaa aatgttggat gctcatactc gtcctttttc aatattattg
     5821 aagcatttat cagggttact agtacgtctc tcaaggataa gtaagtaata ttaaggtacg
     5881 ggaggtattg gacaggccgc aataaaatat ctttattttc attacatctg tgtgttggtt
     5941 ttttgtgtga atcgatagta ctaacatacg ctctccatca aaacaaaacg aaacaaaaca
     6001 aactagcaaa ataggctgtc cccagtgcaa gtgcaggtgc cagaacattt ctct
//
