LOCUS       sequence-306859-        6492 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destabilized firefly luciferase reporter containing the upstream 
            Human PGC-1alpha Distal Promoter from -500 to +52..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6492)
  AUTHORS   Musicant AM, Parag-Sharma K, Gong W, Sengupta M, Chatterjee A, Henry
            EC, Tsai YH, Hayward MC, Sheth S, Betancourt R, Hackman TG, Padilla 
            RJ, Parker JS, Giudice J, Flaveny CA, Hayes DN, Amelio AL
  TITLE     CRTC1/MAML2 directs a PGC-1alpha-IGF-1 circuit that confers 
            vulnerability to PPARgamma inhibition.
  JOURNAL   Cell Rep. 2021 Feb 23;34(8):108768. doi: 
            10.1016/j.celrep.2021.108768.
  PUBMED    33626346
REFERENCE   2  (bases 1 to 6492)
  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: "Cell Rep.";
            date: "2021-02-23"; volume: "34(8):108768. doi"; pages: " 
            10.1016/j.celrep.2021.108768"
COMMENT     Sequence Label: pPGC1a_Distal Promoter-FLuc2P
FEATURES             Location/Qualifiers
     source          1..6492
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        65..256
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     213..233
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    450..571
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(487..506)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     541..560
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(623..742)
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     CDS             complement(746..2395)
                     /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"
     regulatory      2392..2401
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    3018..3109
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(3050..3069)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    3123..3171
                     /note="synthetic polyadenylation signal"
     CDS             join(3276..3344,3345..4136)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4336..4924
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4825..4844
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5078..5095
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    5251..5299
                     /note="synthetic polyadenylation signal"
     CDS             complement(5323..6360)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5336..5344
                     /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)"
     promoter        complement(join(6391..6492,1..256))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6405..6492,1..48)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6459..6478)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag
       61 gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
      121 ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc tttgcatact
      181 tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat tccacagctg
      241 gttctttccg cctcagaagg tagctaacca agttcctctt tcagaggtta tttcaggcca
      301 tggtgctgcg cagatcagcg gaagagcgcc caatacgcaa acggatcctt atcgatttta
      361 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
      421 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
      481 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
      541 gtggtttgtc caaactcatc aatgtatctt atcatgtctg ctcgaagcgg ccggccgccc
      601 cgactctaga gtcgcggcct tagacgttga tcctggcgct ggcgcaagca gcagggtgtc
      661 tatccatgcc gctctcctgg gcgcagctca tgggcagggt gccggcggcc tgctcctcca
      721 cctcgggagg gaagccgtga gaattcacgg cgatcttgcc gcccttcttg gccttaatga
      781 gaatctcgcg gatcttgcgg gcgtccaact tgccggtcag tcctttaggc acctcgtcca
      841 cgaacacaac accaccgcgc agcttcttgg cggttgtaac ctggctggcc acatagtcca
      901 cgatctcctt ctcggtcatg gttttaccgt gttccagcac gacgactgcg gcgggcagct
      961 cgccggcatc gtcgtcgggc aggccggcga ccccggcgtc gaagatgttg gggtgttgca
     1021 gcaggatgct ctccagttcg gctggggcta cctggtagcc cttgtatttg atcaggctct
     1081 tcagccggtc cacgatgaag aagtgctcgt cctcgtccca gtaggcgatg tcgccgctgt
     1141 gcagccagcc gtccttgtcg atgagagcgt ttgtagcctc ggggttgtta acgtagccgc
     1201 tcatgatcat ggggccacgg acgcacagct cgccgcgctg gttcacaccc agtgtcttac
     1261 cggtgtccaa gtccaccacc ttagcctcga agaagggcac caccttgcct actgcgccag
     1321 gcttgtcgtc cccttcgggg gtgatcagaa tggcgctggt tgtttctgtc aggccgtagc
     1381 cctggcggat gcctggtagg tggaagcgtt tggccacggc ctcacctacc tccttgctga
     1441 gcggcgcccc gccgctggcg atctcgtgca agttgcttag gtcgtacttg tcgatgagag
     1501 tgctcttagc gaagaagcta aatagtgtgg gcaccagcag ggcagattga atcttatagt
     1561 cttgcaagct gcgcaagaat agctcctcct cgaagcggta catgagcacg acccgaaagc
     1621 cgcagatcaa gtagcccagc gtggtgaaca tgccgaagcc gtggtgaaat ggcaccacgc
     1681 tgaggatagc ggtgtcgggg atgatctggt tgccgaagat ggggtcgcgg gcatgactga
     1741 atcggacaca agcggtgcgg tgcggtaggg ctacgccctt gggcaatccg gtactgccac
     1801 tactgttcat gatcagggcg atggttttgt cccggtcgaa gctctcgggc acgaagtcgt
     1861 actcgttgaa gccgggtggc aaatgggaag tcacgaaggt gtacatgctt tggaagccct
     1921 ggtagtcggt cttgctatcc atgatgatga tcttttgtat gatcggtagc ttcttttgca
     1981 cgttgaggat cttttgcagc cctttcttgc tcacgaatac gacggtgggc tggctgatgc
     2041 ccatgctgtt cagcagctcg cgctcgttgt agatgtcgtt agctggggcc acagccacac
     2101 cgatgaacag ggcacccaac acgggcatga agaactgcaa gctattctcg ctgcacacca
     2161 cgatccgatg gtttgtattc agcccatagc gcttcatagc ttctgccagc cgaacgctca
     2221 tctcgaagta ctcggcgtag gtaatgtcca cctcgatatg tgcgtcggta aaggcgatgg
     2281 tgccgggcac cagggcgtag cgcttcatgg ctttgtgcag ctgctcgccg gcggtcccgt
     2341 cttcgagtgg gtagaatggc gctgggccct tcttaatgtt tttggcatct tccatggtgg
     2401 ctttaccaac agtaccggat tgccaagctt ctcacaacaa gggcttcggc aggccacttt
     2461 ttaattcaca agggcatggt gacgtgggag ggcatctcag agtcactact aaatgaactc
     2521 cacaataggg cacaaatctg aaacccaatc ttagccagat acgaacgtaa gagcaaatat
     2581 gtacagtatc atgtgctttc cagcagccct gtgtctcgag ggagctctct gacatcagca
     2641 attgagacag gtggttccct ttccaacccg tggaaaataa aaccctagtt cagaggcact
     2701 aagtaaatat agcagtactg taatttttcc ttggtgcact ctccctgtga tttctgtcct
     2761 atgatgacac tttatcccgg gaacccattc aggacatcca tccatgccca taactgactg
     2821 aacatttgca aactggttaa tattgacatt aaagagagtg tagaagactc tgttttatgg
     2881 ccccagttgg tagagctagg gttttcctcc tctcatttat ttggaatacg ttgtcaagtg
     2941 gctgctcaaa actttactgg attatttcac cctttgtccc ctgttgctag cgagctcagg
     3001 taccggccag ttaggccaga gaaatgttct ggcacctgca cttgcactgg ggacagccta
     3061 ttttgctagt ttgttttgtt tcgttttgtt ttgatggaga gcgtatgtta gtactatcga
     3121 ttcacacaaa aaaccaacac acagatgtaa tgaaaataaa gatattttat tgcggcctgt
     3181 ccaatacctc ccgtacctta atattactta cttatccttg agagacgtac tagtaaccct
     3241 gataaatgct tcaataatat tgaaaaagga cgagtatgag catccaacat tttcgtgtcg
     3301 cactcattcc cttctttgcg gcattttgct tgcctgtttt tgcacacccc gaaacgctgg
     3361 tgaaagtaaa agatgctgaa gatcaactgg gtgcaagagt gggctatatc gaactggatc
     3421 tcaatagcgg caagatcctt gagtctttcc gccccgaaga acgattcccg atgatgagca
     3481 ctttcaaagt actgctatgt ggcgcggtgt tgtcccgtat agacgccggg caagagcagc
     3541 ttggtcgccg tatacactac tcacaaaacg acttggttga gtactcgccg gtcacggaaa
     3601 agcatcttac ggatggcatg acggtaagag aattgtgtag tgctgccatt accatgagcg
     3661 acaacaccgc ggccaactta cttctgacaa cgatcggagg ccctaaggag ctgactgcat
     3721 ttcttcataa tatgggtgat catgtgaccc ggcttgaccg ctgggaacca gagttgaacg
     3781 aagccatacc gaacgacgag cgtgatacca cgatgccagt agcaatggcc acaactcttc
     3841 ggaaactact cactggcgaa cttcttactc tagcatcacg acagcagctc atagactgga
     3901 tggaggcgga caaagtagca ggaccacttc ttcgctcggc cctccctgct ggctggttca
     3961 ttgctgacaa atcgggggcc ggtgaacgcg gctctcgcgg catcattgct gcgctggggc
     4021 ctgatggtaa gccctcacga atcgtagtga tctacacgac ggggagtcag gccactatgg
     4081 acgaacgaaa taggcagatc gctgagatcg gtgcctcact gatcaagcac tggtaaccac
     4141 tgcagtggtt tagcatttgc ggccgctgtc agaccaagtt tactcatata tactttagat
     4201 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     4261 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     4321 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4381 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     4441 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     4501 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     4561 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4621 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4681 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4741 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4801 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4861 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4921 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4981 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     5041 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     5101 ggaagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg
     5161 cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca
     5221 agggcatcgg tcgacagcat ctagttcgaa cacacaaaaa accaacacac agatgtaatg
     5281 aaaataaaga tattttattg cggccatgac cggttctaga gtttaaactc gacctacctc
     5341 cttagcgcgc ggtcgtgtac tgggccggcg gttgccgctg tcggccagca cctcgacgca
     5401 gccgtcggtc catacggctg cgctccggcg agcgatttgt gtgcgaccga cggtgccggc
     5461 cccgctgcgg acgatggcat cgcagcggcc ttgagcccag gcagcatcgt cgaagttgcc
     5521 gtccacgagg ctctggtaga gctgatccag gccgatgcgc agcatgtagg ctcgcagacg
     5581 agggctgccg gccagctcgg gatgccggcg ctcgaagtag cgagtctgct gctccatgca
     5641 agccagccag ggccgccaga agaagatgtt ggccacctcg tactgactgt ccccgaacat
     5701 agcttcggac cagtcgatta cggcggtgat gcggccgttg tcggtcagga cgttgttgct
     5761 gccgaagtcg gcatggacga ggtggcgcac ctcgggacag tcttcggccc acagcatcag
     5821 ttcgtccagg gcttgagcta cgctggcgga cacggtgtcg tccatcacgg tctgccagtg
     5881 gtagacatgg ggatcagcaa tggcgcaaat gaaatcccgc caggttgtgt actggccgat
     5941 gccttggggc ccgaatgggc cgaagccgct ggtttgactg aggtcggcgg cggcaatagc
     6001 atccatagct tcggcgacag gctgtaacac agcaggcagc tctgtttcgg ggaggtcttg
     6061 gagagtaacg ccttgggcgc gtctactgat gcagtatgtc aggctctcgc tgaactcgcc
     6121 gatgtccaac acttcgggga tgggtagtgc agcgctggcg aagtggcggt acacgtagcg
     6181 gtctttgtag aagccatcag cgcagctatt cacccgcagt acatagccgc gtccgccgac
     6241 atcgaagctg aaggctcggc tctcttcgcc ctccgacaac tgcatcaggt cgctcacact
     6301 gtcgaacttc tcgatgagaa atttttcaac gctggtagcg gtgagttcgg gcttcttcat
     6361 ggtggcgcta gtgtcagaag aatcgagctt tttgcaaaag cctaggcctc caaaaaagcc
     6421 tcctcactac ttctggaata gctcagaggc agaggcggcc tcggcctctg cataaataaa
     6481 aaaaattagt ca
//
