LOCUS       sequence-306858-        6551 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Destabilized firefly luciferase reporter containing the Human 
            PGC-1alpha Proximal Promoter from -500 to +52..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6551)
  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 6551)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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_Proximal Promoter-FLuc2P
FEATURES             Location/Qualifiers
     source          1..6551
                     /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    3077..3168
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(3109..3128)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    3182..3230
                     /note="synthetic polyadenylation signal"
     CDS             join(3335..3403,3404..4195)
                     /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      4395..4983
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4884..4903
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5137..5154
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    5310..5358
                     /note="synthetic polyadenylation signal"
     CDS             complement(5382..6419)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5395..5403
                     /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(6450..6551,1..256))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6464..6551,1..48)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6518..6537)
                     /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 tttcaactcc aatccacagt gacacagagc
     2461 acacactcat gcaggcaacc agccccttac tgagagtgaa ctgaaggcac ctgtcttact
     2521 acagtcccca gtcacatgac aaagctatta aaaagtaggc tgggctgtca ctcacccagc
     2581 ctcccttccc ctgtgcagct tgctgctcta actcgtgacg tcactcaaag gcagccctct
     2641 gcttcagtga agtaacgctt taaaaaaaaa aaaagaaaga aaaagaaaag aaagaaagaa
     2701 aggaaacact tagacttttg gaggcttcaa gcatcatgct gtgattaaag ccagctttga
     2761 atgccacaga ctctaaacgg agccctggcc atataataac caaaatcccc tgcaatcttt
     2821 ttttaattta ttttccttcc aaaacaagca agtggcaaag ctccctgttt catgacagag
     2881 taactatatt cttagctaaa acgtgtattc cctaagaact ctttaattct aagatctcca
     2941 agtggactca agctcagttt gggactaaag caagagctta tcacatgatg catttttatg
     3001 agacattttc tcattgaggg agtgtttgaa agcgcaaacg gggcgctagc gagctcaggt
     3061 accggccagt taggccagag aaatgttctg gcacctgcac ttgcactggg gacagcctat
     3121 tttgctagtt tgttttgttt cgttttgttt tgatggagag cgtatgttag tactatcgat
     3181 tcacacaaaa aaccaacaca cagatgtaat gaaaataaag atattttatt gcggcctgtc
     3241 caatacctcc cgtaccttaa tattacttac ttatccttga gagacgtact agtaaccctg
     3301 ataaatgctt caataatatt gaaaaaggac gagtatgagc atccaacatt ttcgtgtcgc
     3361 actcattccc ttctttgcgg cattttgctt gcctgttttt gcacaccccg aaacgctggt
     3421 gaaagtaaaa gatgctgaag atcaactggg tgcaagagtg ggctatatcg aactggatct
     3481 caatagcggc aagatccttg agtctttccg ccccgaagaa cgattcccga tgatgagcac
     3541 tttcaaagta ctgctatgtg gcgcggtgtt gtcccgtata gacgccgggc aagagcagct
     3601 tggtcgccgt atacactact cacaaaacga cttggttgag tactcgccgg tcacggaaaa
     3661 gcatcttacg gatggcatga cggtaagaga attgtgtagt gctgccatta ccatgagcga
     3721 caacaccgcg gccaacttac ttctgacaac gatcggaggc cctaaggagc tgactgcatt
     3781 tcttcataat atgggtgatc atgtgacccg gcttgaccgc tgggaaccag agttgaacga
     3841 agccataccg aacgacgagc gtgataccac gatgccagta gcaatggcca caactcttcg
     3901 gaaactactc actggcgaac ttcttactct agcatcacga cagcagctca tagactggat
     3961 ggaggcggac aaagtagcag gaccacttct tcgctcggcc ctccctgctg gctggttcat
     4021 tgctgacaaa tcgggggccg gtgaacgcgg ctctcgcggc atcattgctg cgctggggcc
     4081 tgatggtaag ccctcacgaa tcgtagtgat ctacacgacg gggagtcagg ccactatgga
     4141 cgaacgaaat aggcagatcg ctgagatcgg tgcctcactg atcaagcact ggtaaccact
     4201 gcagtggttt agcatttgcg gccgctgtca gaccaagttt actcatatat actttagatt
     4261 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     4321 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     4381 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     4441 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     4501 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     4561 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     4621 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     4681 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     4741 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     4801 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     4861 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     4921 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     4981 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     5041 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     5101 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     5161 gaagagcgct gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc
     5221 ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa
     5281 gggcatcggt cgacagcatc tagttcgaac acacaaaaaa ccaacacaca gatgtaatga
     5341 aaataaagat attttattgc ggccatgacc ggttctagag tttaaactcg acctacctcc
     5401 ttagcgcgcg gtcgtgtact gggccggcgg ttgccgctgt cggccagcac ctcgacgcag
     5461 ccgtcggtcc atacggctgc gctccggcga gcgatttgtg tgcgaccgac ggtgccggcc
     5521 ccgctgcgga cgatggcatc gcagcggcct tgagcccagg cagcatcgtc gaagttgccg
     5581 tccacgaggc tctggtagag ctgatccagg ccgatgcgca gcatgtaggc tcgcagacga
     5641 gggctgccgg ccagctcggg atgccggcgc tcgaagtagc gagtctgctg ctccatgcaa
     5701 gccagccagg gccgccagaa gaagatgttg gccacctcgt actgactgtc cccgaacata
     5761 gcttcggacc agtcgattac ggcggtgatg cggccgttgt cggtcaggac gttgttgctg
     5821 ccgaagtcgg catggacgag gtggcgcacc tcgggacagt cttcggccca cagcatcagt
     5881 tcgtccaggg cttgagctac gctggcggac acggtgtcgt ccatcacggt ctgccagtgg
     5941 tagacatggg gatcagcaat ggcgcaaatg aaatcccgcc aggttgtgta ctggccgatg
     6001 ccttggggcc cgaatgggcc gaagccgctg gtttgactga ggtcggcggc ggcaatagca
     6061 tccatagctt cggcgacagg ctgtaacaca gcaggcagct ctgtttcggg gaggtcttgg
     6121 agagtaacgc cttgggcgcg tctactgatg cagtatgtca ggctctcgct gaactcgccg
     6181 atgtccaaca cttcggggat gggtagtgca gcgctggcga agtggcggta cacgtagcgg
     6241 tctttgtaga agccatcagc gcagctattc acccgcagta catagccgcg tccgccgaca
     6301 tcgaagctga aggctcggct ctcttcgccc tccgacaact gcatcaggtc gctcacactg
     6361 tcgaacttct cgatgagaaa tttttcaacg ctggtagcgg tgagttcggg cttcttcatg
     6421 gtggcgctag tgtcagaaga atcgagcttt ttgcaaaagc ctaggcctcc aaaaaagcct
     6481 cctcactact tctggaatag ctcagaggca gaggcggcct cggcctctgc ataaataaaa
     6541 aaaattagtc a
//
