LOCUS       sequence-306860-        8036 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destabilized firefly luciferase reporter containing the canonical 
            Human IGF-1 P1 Promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8036)
  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 8036)
  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: pIGF1_P1 Promoter-FLuc2P
FEATURES             Location/Qualifiers
     source          1..8036
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(41..60)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        139..330
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     287..307
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    524..645
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(561..580)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     615..634
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(697..816)
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     CDS             complement(820..2469)
                     /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      2466..2475
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    4636..4727
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(4668..4687)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    4741..4789
                     /note="synthetic polyadenylation signal"
     CDS             join(4894..4962,4963..5754)
                     /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      5954..6542
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6443..6462
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6696..6713
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6869..6917
                     /note="synthetic polyadenylation signal"
     CDS             complement(6941..7978)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             6954..6962
                     /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(8009..8036,1..330))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(8023..8036,1..122)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cctcctcact acttctggaa tagctcagag gcagaggcgg cctcggcctc tgcataaata
       61 aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg
      121 atgggcggag ttaggggcgg gactatggtt gctgactaat tgagatgcat gctttgcata
      181 cttctgcctg ctggggagcc tggggacttt ccacacctgg ttgctgacta attgagatgc
      241 atgctttgca tacttctgcc tgctggggag cctggggact ttccacaccc taactgacac
      301 acattccaca gctggttctt tccgcctcag aaggtagcta accaagttcc tctttcagag
      361 gttatttcag gccatggtgc tgcgcagatc agcggaagag cgcccaatac gcaaacggat
      421 ccttatcgat tttaccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
      481 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
      541 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
      601 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgctcgaa
      661 gcggccggcc gccccgactc tagagtcgcg gccttagacg ttgatcctgg cgctggcgca
      721 agcagcaggg tgtctatcca tgccgctctc ctgggcgcag ctcatgggca gggtgccggc
      781 ggcctgctcc tccacctcgg gagggaagcc gtgagaattc acggcgatct tgccgccctt
      841 cttggcctta atgagaatct cgcggatctt gcgggcgtcc aacttgccgg tcagtccttt
      901 aggcacctcg tccacgaaca caacaccacc gcgcagcttc ttggcggttg taacctggct
      961 ggccacatag tccacgatct ccttctcggt catggtttta ccgtgttcca gcacgacgac
     1021 tgcggcgggc agctcgccgg catcgtcgtc gggcaggccg gcgaccccgg cgtcgaagat
     1081 gttggggtgt tgcagcagga tgctctccag ttcggctggg gctacctggt agcccttgta
     1141 tttgatcagg ctcttcagcc ggtccacgat gaagaagtgc tcgtcctcgt cccagtaggc
     1201 gatgtcgccg ctgtgcagcc agccgtcctt gtcgatgaga gcgtttgtag cctcggggtt
     1261 gttaacgtag ccgctcatga tcatggggcc acggacgcac agctcgccgc gctggttcac
     1321 acccagtgtc ttaccggtgt ccaagtccac caccttagcc tcgaagaagg gcaccacctt
     1381 gcctactgcg ccaggcttgt cgtccccttc gggggtgatc agaatggcgc tggttgtttc
     1441 tgtcaggccg tagccctggc ggatgcctgg taggtggaag cgtttggcca cggcctcacc
     1501 tacctccttg ctgagcggcg ccccgccgct ggcgatctcg tgcaagttgc ttaggtcgta
     1561 cttgtcgatg agagtgctct tagcgaagaa gctaaatagt gtgggcacca gcagggcaga
     1621 ttgaatctta tagtcttgca agctgcgcaa gaatagctcc tcctcgaagc ggtacatgag
     1681 cacgacccga aagccgcaga tcaagtagcc cagcgtggtg aacatgccga agccgtggtg
     1741 aaatggcacc acgctgagga tagcggtgtc ggggatgatc tggttgccga agatggggtc
     1801 gcgggcatga ctgaatcgga cacaagcggt gcggtgcggt agggctacgc ccttgggcaa
     1861 tccggtactg ccactactgt tcatgatcag ggcgatggtt ttgtcccggt cgaagctctc
     1921 gggcacgaag tcgtactcgt tgaagccggg tggcaaatgg gaagtcacga aggtgtacat
     1981 gctttggaag ccctggtagt cggtcttgct atccatgatg atgatctttt gtatgatcgg
     2041 tagcttcttt tgcacgttga ggatcttttg cagccctttc ttgctcacga atacgacggt
     2101 gggctggctg atgcccatgc tgttcagcag ctcgcgctcg ttgtagatgt cgttagctgg
     2161 ggccacagcc acaccgatga acagggcacc caacacgggc atgaagaact gcaagctatt
     2221 ctcgctgcac accacgatcc gatggtttgt attcagccca tagcgcttca tagcttctgc
     2281 cagccgaacg ctcatctcga agtactcggc gtaggtaatg tccacctcga tatgtgcgtc
     2341 ggtaaaggcg atggtgccgg gcaccagggc gtagcgcttc atggctttgt gcagctgctc
     2401 gccggcggtc ccgtcttcga gtgggtagaa tggcgctggg cccttcttaa tgtttttggc
     2461 atcttccatg gtggctttac caacagtacc ggattgccaa gcttctaggc agaagcaaac
     2521 agtacacaat ttaaatagaa ttccccaatg acttcaaaga gtaagaaata tttaccttca
     2581 agaaatcaca aaagcagcac ttaaataatt gggttggaag actgctgatt tttcccattg
     2641 cttctgaagt acaaagtctg aaaatgaatt ggttagcagg aataatgaag caaaaagaaa
     2701 tccagagaga tgggagatgt tgagagcaat gtcacatttc aattttgagg actttattcc
     2761 attgcgcagg ctctatctgc tctgaattta gcagtgacag tgagatttag caaacagaag
     2821 agggatttag agaaaatcct cacatttatc tacaaaacac agacactgta gacaggaaac
     2881 agctggggga acatttgcct tctctctctc tccctcttct ggcaaagtta ttgagtaagg
     2941 acttttttgg gcatggtgac aaataacatc atacctttgc attttaaaac tagagcacag
     3001 aagcattttt ttcccttaaa agaatgtgtg ttagtgacag ggttcgcaga cattaaaata
     3061 cttatgctgc catagaaaat aaggatctgt tttctgatta actttctgct gggcatgaag
     3121 acacaaacgt ctgctaatac accttaccag tattaaagga atatggggga tgggagagca
     3181 attttagatt ctagagcaaa tgcattattt aaaacaaaaa aagaaagaaa ggacgataag
     3241 accctctcca gcaaactcta caattttaac cctgaagtga ctggggtaaa gtagattgga
     3301 agacagcact cgggtgaccc cttgtcccag ttgccaagtg agaggtgtga tctcatttcc
     3361 tagaacctat ggaggggcag gcagcattgg attggtccct ttagggcact gtctcatact
     3421 ttttctcctg cgatgaggca aagactatgc cgagctgtaa aacccaactg aaatagaaca
     3481 tttttcttta ttcttctact aggctgcctg tcactgtccc ctccccaccc caccactctg
     3541 ggagaagggt acctgggggg caaacaattt tcctgttgtt tgaattttcc atataactca
     3601 aacctgtgtg tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg tgtgtatgac agagggagtc
     3661 tgtgtgccag agtgtgtttt ctttctctga tttttgtgta ttctaaataa caccagctgg
     3721 ctagcaatac cctctcgagg gttatgatgt cattcaaatc cctcaactgg gtgactggcc
     3781 ggcaacatgg tccaagccag ctgttttgct atttataatg tacacttgcc tttgccattg
     3841 agaattcagt tctgatcttt tgtcgataca gtttgtacct atttcccttt ggttttgtga
     3901 agcagtagat aagaaagtga attataaaat aaggttgcaa aagcccagag cagacatacc
     3961 tctttcccta gagagctctc caggcctggt ttcccaggag tggtggaaat aacctggacc
     4021 ttgaattttt tctttttttt ttttttccac atgactctca ggggactgac acatcaactg
     4081 aaaacacagt tctgcttgaa atcctactct ggctctttgt gttatcctgg ggccagaaat
     4141 ttgtttgtag aagtgggtgc cctaacaaaa aacaaacaaa cctgctttgt ttggaagggg
     4201 gttgggtttg tttgagtctt ctgtgtggtt aatacattgt agcaatgttt acctttcaga
     4261 aatcaaagtt agaaatctgg agttatgttg tgtcaatggt gcactatgtt cagcaacatt
     4321 ttgtgagctc tactatgtgc ccaactacaa catccctagg aaaacaaaga ggaataagat
     4381 atggtcaagt cccttcaggg gctttcagct gaatgtcaga gacaaagagg aaaacttcac
     4441 agtcatatga tgggcatgtg ctcttttaaa aacatgaaca gtgcacaatg agagggagtg
     4501 acacattctc tgggaagaca ggggagtaaa atcttttgga aaatccaaaa tgagggaggc
     4561 agattgtgtc atgagaggct ttggagatct tgatatcctc gaggctagcg agctcaggta
     4621 ccggccagtt aggccagaga aatgttctgg cacctgcact tgcactgggg acagcctatt
     4681 ttgctagttt gttttgtttc gttttgtttt gatggagagc gtatgttagt actatcgatt
     4741 cacacaaaaa accaacacac agatgtaatg aaaataaaga tattttattg cggcctgtcc
     4801 aatacctccc gtaccttaat attacttact tatccttgag agacgtacta gtaaccctga
     4861 taaatgcttc aataatattg aaaaaggacg agtatgagca tccaacattt tcgtgtcgca
     4921 ctcattccct tctttgcggc attttgcttg cctgtttttg cacaccccga aacgctggtg
     4981 aaagtaaaag atgctgaaga tcaactgggt gcaagagtgg gctatatcga actggatctc
     5041 aatagcggca agatccttga gtctttccgc cccgaagaac gattcccgat gatgagcact
     5101 ttcaaagtac tgctatgtgg cgcggtgttg tcccgtatag acgccgggca agagcagctt
     5161 ggtcgccgta tacactactc acaaaacgac ttggttgagt actcgccggt cacggaaaag
     5221 catcttacgg atggcatgac ggtaagagaa ttgtgtagtg ctgccattac catgagcgac
     5281 aacaccgcgg ccaacttact tctgacaacg atcggaggcc ctaaggagct gactgcattt
     5341 cttcataata tgggtgatca tgtgacccgg cttgaccgct gggaaccaga gttgaacgaa
     5401 gccataccga acgacgagcg tgataccacg atgccagtag caatggccac aactcttcgg
     5461 aaactactca ctggcgaact tcttactcta gcatcacgac agcagctcat agactggatg
     5521 gaggcggaca aagtagcagg accacttctt cgctcggccc tccctgctgg ctggttcatt
     5581 gctgacaaat cgggggccgg tgaacgcggc tctcgcggca tcattgctgc gctggggcct
     5641 gatggtaagc cctcacgaat cgtagtgatc tacacgacgg ggagtcaggc cactatggac
     5701 gaacgaaata ggcagatcgc tgagatcggt gcctcactga tcaagcactg gtaaccactg
     5761 cagtggttta gcatttgcgg ccgctgtcag accaagttta ctcatatata ctttagattg
     5821 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5881 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5941 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     6001 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     6061 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6121 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6181 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6241 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6301 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6361 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6421 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6481 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6541 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6601 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     6661 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     6721 aagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg
     6781 gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag
     6841 ggcatcggtc gacagcatct agttcgaaca cacaaaaaac caacacacag atgtaatgaa
     6901 aataaagata ttttattgcg gccatgaccg gttctagagt ttaaactcga cctacctcct
     6961 tagcgcgcgg tcgtgtactg ggccggcggt tgccgctgtc ggccagcacc tcgacgcagc
     7021 cgtcggtcca tacggctgcg ctccggcgag cgatttgtgt gcgaccgacg gtgccggccc
     7081 cgctgcggac gatggcatcg cagcggcctt gagcccaggc agcatcgtcg aagttgccgt
     7141 ccacgaggct ctggtagagc tgatccaggc cgatgcgcag catgtaggct cgcagacgag
     7201 ggctgccggc cagctcggga tgccggcgct cgaagtagcg agtctgctgc tccatgcaag
     7261 ccagccaggg ccgccagaag aagatgttgg ccacctcgta ctgactgtcc ccgaacatag
     7321 cttcggacca gtcgattacg gcggtgatgc ggccgttgtc ggtcaggacg ttgttgctgc
     7381 cgaagtcggc atggacgagg tggcgcacct cgggacagtc ttcggcccac agcatcagtt
     7441 cgtccagggc ttgagctacg ctggcggaca cggtgtcgtc catcacggtc tgccagtggt
     7501 agacatgggg atcagcaatg gcgcaaatga aatcccgcca ggttgtgtac tggccgatgc
     7561 cttggggccc gaatgggccg aagccgctgg tttgactgag gtcggcggcg gcaatagcat
     7621 ccatagcttc ggcgacaggc tgtaacacag caggcagctc tgtttcgggg aggtcttgga
     7681 gagtaacgcc ttgggcgcgt ctactgatgc agtatgtcag gctctcgctg aactcgccga
     7741 tgtccaacac ttcggggatg ggtagtgcag cgctggcgaa gtggcggtac acgtagcggt
     7801 ctttgtagaa gccatcagcg cagctattca cccgcagtac atagccgcgt ccgccgacat
     7861 cgaagctgaa ggctcggctc tcttcgccct ccgacaactg catcaggtcg ctcacactgt
     7921 cgaacttctc gatgagaaat ttttcaacgc tggtagcggt gagttcgggc ttcttcatgg
     7981 tggcgctagt gtcagaagaa tcgagctttt tgcaaaagcc taggcctcca aaaaag
//
