LOCUS       sequence-341222-        5665 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To overexpress PPARgamma1 in Mammalian Cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5665)
  AUTHORS   Kim JH, Park KW, Lee EW, Jang WS, Seo J, Shin S, Hwang KA, Song J
  TITLE     Suppression of PPARgamma through MKRN1-mediated ubiquitination and 
            degradation prevents adipocyte differentiation.
  JOURNAL   Cell Death Differ. 2014 Apr;21(4):594-603. doi: 
            10.1038/cdd.2013.181. Epub 2013 Dec 13.
  PUBMED    24336050
REFERENCE   2  (bases 1 to 5665)
  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: 
            "10.1038/cdd.2013.181"; journalName: "Cell Death Differ"; date: 
            "2014-04"; volume: "21"; issue: "4"; pages: "594-603"
COMMENT     Sequence Label: PCS4 3XFLAG-PPARgamma1
FEATURES             Location/Qualifiers
     source          1..5665
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..978
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1013..1031
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1013..1030
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1107..1130
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1134..1157
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1161..1184
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2671..2690)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2672..2690)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2695..2829
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2719..2738)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2773..2792
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2942..2962)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2942..2960)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2981..2997)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2981..2997)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2994..3016)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3005..3021
                     /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)."
     promoter        complement(join(3029..3035,3036..3053,3054..3059))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3074..3095
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3212..3229)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3383..3971)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3463..3482)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4142..4933,4934..5002))
                     /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     4765..4784
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5003..5107)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5133..5588)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5270..5291)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5482..5501
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
       61 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
      121 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt
      181 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg
      241 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
      301 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
      361 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
      421 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
      481 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
      541 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
      601 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
      661 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
      721 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
      781 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
      841 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
      901 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
      961 aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct tgatttaggt
     1021 gacactatag aatacaagct acttgttctt tttgcaggat cccatcgatt gtacagatat
     1081 ctggcgcgcc taccatgggg cccggcgact acaaggatga cgatgacaag ggcgactaca
     1141 aggatgacga tgacaagggc gactacaagg atgacgatga caagggccgg cctaagatct
     1201 ggaattcaag gcctctcgag atgaccatgg ttgacacaga gatgccattc tggcccacca
     1261 acttcggaat cagctctgtg gacctctccg tgatggaaga ccactcgcat tcctttgaca
     1321 tcaagccctt taccacagtt gatttctcca gcatttctgc tccacactat gaagacattc
     1381 cattcacaag agctgaccca atggttgctg attacaaata tgacctgaag ctccaagaat
     1441 accaaagtgc gatcaaagta gaacctgcat ctccacctta ttattctgaa aagacccagc
     1501 tctacaacag gcctcatgaa gaaccttcta actccctcat ggccattgag tgccgagtct
     1561 gtggggataa agcatcaggc ttccactatg gagttcatgc ttgtgaagga tgcaagggtt
     1621 ttttccgaag aaccatccga ttgaagctta tttatgatag gtgtgatctt aactgccgga
     1681 tccacaaaaa aagtagaaat aaatgtcagt actgtcggtt tcagaagtgc cttgctgtgg
     1741 ggatgtctca caatgccatc aggtttgggc ggatgccaca ggccgagaag gagaagctgt
     1801 tggcggagat ctccagtgat atcgaccagc tgaacccaga gtctgctgat ctgcgagccc
     1861 tggcaaagca tttgtatgac tcatacataa agtccttccc gctgaccaaa gccaaggcga
     1921 gggcgatctt gacaggaaag acaacggaca aatcaccatt tgtcatctac gacatgaatt
     1981 ccttaatgat gggagaagat aaaatcaagt tcaaacatat cacccccctg caggagcaga
     2041 gcaaagaggt ggccatccga atttttcaag ggtgccagtt tcgatccgta gaagccgtgc
     2101 aagagatcac agagtatgcc aaaaatatcc ctggtttcat taaccttgat ttgaatgacc
     2161 aagtgactct gctcaagtat ggtgtccatg agatcatcta cacgatgctg gcctccctga
     2221 tgaataaaga tggagtcctc atctcagagg gccaaggatt catgaccagg gagttcctca
     2281 aaagcctgcg gaagcccttt ggtgacttta tggagcctaa gtttgagttt gctgtgaagt
     2341 tcaatgcact ggaattagat gacagtgact tggctatatt tatagctgtc attattctca
     2401 gtggagaccg cccaggcttg ctgaacgtga agcccatcga ggacatccaa gacaacctgc
     2461 tgcaggccct ggaactgcag ctcaagctga atcacccaga gtcctctcag ctgttcgcca
     2521 aggtgctcca gaagatgaca gacctcaggc agatcgtcac agagcacgtg cagctactgc
     2581 atgtgatcaa gaagacagag acagacatga gccttcaccc cctgctccag gagatctaca
     2641 aggacttgta ctagtctaga tagctagcct ccctatagtg agtcgtatta cgtagatcca
     2701 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     2761 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     2821 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
     2881 gaggtttttt aattcgcggc cgcggcgcca atgcattggg cccggtaccc agcttttgtt
     2941 ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt
     3001 gaaattgtta tccgctcaca attccacaca acatacgagc cgggagcata aagtgtaaag
     3061 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     3121 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     3181 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     3241 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     3301 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3361 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     3421 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     3481 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3541 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     3601 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     3661 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     3721 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     3781 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     3841 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     3901 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     3961 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4021 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4081 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4141 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4201 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     4261 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4321 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     4381 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4441 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     4501 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     4561 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     4621 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     4681 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     4741 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     4801 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     4861 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     4921 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     4981 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     5041 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5101 ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa
     5161 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     5221 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     5281 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     5341 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     5401 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     5461 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     5521 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     5581 gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     5641 ctcttcgcta ttacgccagt cgacc
//
