LOCUS       sequence-435431-        6763 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PPAR reporter.  3xDR1 sites upstream of a luciferase reporter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6763)
  AUTHORS   Kim JB, Wright HM, Wright M, Spiegelman BM
  TITLE     ADD1/SREBP1 activates PPARgamma through the production of endogenous
            ligand.
  JOURNAL   Proc Natl Acad Sci U S A 1998 Apr 14;95(8):4333-7.
  PUBMED    9539737
REFERENCE   2  (bases 1 to 6763)
  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: "Proc Natl 
            Acad Sci U S A"; date: "1998-04-14"; volume: "95(8)"; pages: 
            "4333-7"
COMMENT     Sequence Label: PPRE X3-TK-luc
FEATURES             Location/Qualifiers
     source          1..6763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(274..291)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(274..290)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(283..305)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      731..1244
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(818..837)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1028..1049
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1726..1745)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1905..1923)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1991..2095
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2096..2164,2165..2956)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2314..2333)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3127..3715
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3616..3635
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3869..3886
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4003..4024
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4039..4044,4045..4062,4063..4069)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4077..4093
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     4082..4104
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4101..4117
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4101..4117
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    4135..4269
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4159..4178)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4213..4232
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4694..4714)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          4844..4909
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(5040..6692)
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     6608..6626
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     promoter        complement(join(6752..6763,1..51))
                     /label=Mini-TK promoter
                     /note="minimal herpes simplex virus (HSV) thymidine kinase 
                     promoter"
ORIGIN
        1 ctgcagggtc gctcggtgtt cgaggccaca cgcgtcacct taatatgcga agtggacctc
       61 ggaccgcgcc gccccgactg catctgcgtg ttcgaattcg ccaatgacaa gacgctgggc
      121 ggggccggat cctctagagt cgactcccga acgtgacctt tgtcctggtc ccctgtcgac
      181 tcccgaacgt gacctttgtc ctggtcccct gtcgactccc gaacgtgacc tttgtcctgg
      241 tcccctgtcg acctgcaggc atgcaagctt ggcactggcc gtcgttttac aacgtcgtga
      301 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
      361 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
      421 tggcgaatgg cgattccgtt gcaatggctg gcggtaatat tgttctggat attaccagca
      481 aggccgatag tttgagttct tctactcagg caagtgatgt tattactaat caaagaagta
      541 ttgcgacaac ggttaatttg cgtgatggac agactctttt actcggtggc ctcactgatt
      601 ataaaaacac ttctcaggat tctggcgtac cgttcctgtc taaaatccct ttaatcggcc
      661 tcctgtttag ctcccgctct gattctaacg aggaaagcac gttatacgtg ctcgtcaaag
      721 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
      781 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
      841 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
      901 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
      961 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     1021 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     1081 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     1141 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaaat atttgcttat
     1201 acaatcttcc tgtttttggg gcttttctga ttatcaaccg gggtacatat gattgacatg
     1261 ctagttttac gattaccgtt catcgattct cttgtttgct ccagactctc aggcaatgac
     1321 ctgatagcct ttgtagagac ctctcaaaaa tagctaccct ctccggcatg aatttatcag
     1381 ctagaacggt tgaatatcat attgatggtg atttgactgt ctccggcctt tctcacccgt
     1441 ttgaatcttt acctacacat tactcaggca ttgcatttaa aatatatgag ggttctaaaa
     1501 atttttatcc ttgcgttgaa ataaaggctt ctcccgcaaa agtattacag ggtcataatg
     1561 tttttggtac aaccgattta gctttatgct ctgaggcttt attgcttaat tttgctaatt
     1621 ctttgccttg cctgtatgat ttattggatg ttggaatcgc ctgatgcggt attttctcct
     1681 tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga
     1741 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     1801 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     1861 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     1921 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     1981 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     2041 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     2101 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     2161 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     2221 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     2281 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     2341 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     2401 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     2461 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     2521 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     2581 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     2641 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     2701 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     2761 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     2821 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     2881 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     2941 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     3001 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     3061 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3121 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3181 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3241 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3301 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3361 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3421 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3481 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3541 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3601 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3661 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3721 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     3781 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     3841 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     3901 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     3961 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     4021 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     4081 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgaat tcccgatcca
     4141 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     4201 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     4261 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
     4321 gaggtttttt aaagcaagta aaacctctac aaatgtggta tggctgatta tgatctctag
     4381 tcaaggcact atacatcaaa tattccttat taaccccttt acaaattaaa aagctaaagg
     4441 tacacaattt ttgagcatag ttattaatag cagacactct atgcctgtgt ggagtaagaa
     4501 aaaacagtat gttatgatta taactgttat gcctacttat aaaggttaca gaatattttt
     4561 ccataatttt cttgtatagc agtgcagctt tttcctttgt ggtgtaaata gcaaagcaag
     4621 caagagttct attactaaac acagcatgac tcaaaaaact tagcaattct gaaggaaagt
     4681 ccttggggtc ttctaccttt ctcttctttt ttggaggagt agaatgttga gagtcagcag
     4741 tagcctcatc atcactagat ggcatttctt ctgagcaaaa caggttttcc tcattaaagg
     4801 cattccacca ctgctcccat tcatcagttc cataggttgg aatctaaaat acacaaacaa
     4861 ttagaatcag tagtttaaca cattatacac ttaaaaattt tatatttacc ttagagcttt
     4921 aaatctctgt aggtagtttg tccaattatg tcacaccaca gaagtaaggt tccttcacaa
     4981 agatcctcta gagtattaca atagctaaga atttcgtcat cgctgaatac agttacattt
     5041 tacaatttgg actttccgcc cttcttggcc tttatgagga tctctctgat ttttcttgcg
     5101 tcgagttttc cggtaagacc tttcggtact tcgtccacaa acacaactcc tccgcgcaac
     5161 tttttcgcgg ttgttacttg actggcgacg taatccacga tctctttttc cgtcatcgtc
     5221 tttccgtgct ccaaaacaac aacggcggcg ggaagttcac cggcgtcatc gtcgggaaga
     5281 cctgccacgc ccgcgtcgaa gatgttgggg tgttgtaaca atatcgattc caattcagcg
     5341 ggggccacct gatatccttt gtatttaatt aaagacttca agcggtcaac tatgaagaag
     5401 tgttcgtctt cgtcccagta agctatgtct ccagaatgta gccatccatc cttgtcaatc
     5461 aaggcgttgg tcgcttccgg attgtttaca taaccggaca taatcatagg tcctctgaca
     5521 cataattcgc ctctctgatt aacgcccagc gttttcccgg tatccagatc cacaaccttc
     5581 gcttcaaaaa atggaacaac tttaccgacc gcgcccggtt tatcatcccc ctcgggtgta
     5641 atcagaatag ctgatgtagt ctcagtgagc ccatatcctt gtcgtatccc tggaagatgg
     5701 aagcgttttg caaccgcttc cccgacttct ttcgaaagag gtgcgccccc agaagcaatt
     5761 tcgtgtaaat tagataaatc gtatttgtca atcagagtgc ttttggcgaa gaatgaaaat
     5821 agggttggta ctagcaacgc actttgaatt ttgtaatcct gaagggatcg taaaaacagc
     5881 tcttcttcaa atctatacat taagacgact cgaaatccac atatcaaata tccgagtgta
     5941 gtaaacattc caaaaccgtg atggaatgga acaacactta aaatcgcagt atccggaatg
     6001 atttgattgc caaaaatagg atctctggca tgcgagaatc tgacgcaggc agttctatgc
     6061 ggaagggcca cacccttagg taacccagta gatccagagg aattcattat cagtgcaatt
     6121 gttttgtcac gatcaaagga ctctggtaca aaatcgtatt cattaaaacc gggaggtaga
     6181 tgagatgtga cgaacgtgta catcgactga aatccctggt aatccgtttt agaatccatg
     6241 ataataattt tctggattat tggtaatttt ttttgcacgt tcaaaatttt ttgcaacccc
     6301 tttttggaaa caaacactac ggtaggctgc gaaatgttca tactgttgag caattcacgt
     6361 tcattataaa tgtcgttcgc gggcgcaact gcaactccga taaataacgc gcccaacacc
     6421 ggcataaaga attgaagaga gttttcactg catacgacga ttctgtgatt tgtattcagc
     6481 ccatatcgtt tcatagcttc tgccaaccga acggacattt cgaagtattc cgcgtacgtg
     6541 atgttcacct cgatatgtgc atctgtaaaa gcaattgttc caggaaccag ggcgtatctc
     6601 ttcatagcct tatgcagttg ctctccagcg gttccatcct ctagaggata gaatggcgcc
     6661 gggcctttct ttatgttttt ggcgtcttcc attttaccaa cagtaccgga atgccaagct
     6721 cctcgagatc tgcggcacgc tgttgacgct gttaagcggg tcg
//
