LOCUS       sequence-217443-        7042 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of flag-tagged PPAR gamma.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7042)
  AUTHORS   Hauser S, Adelmant G, Sarraf P, Wright HM, Mueller E, Spiegelman BM
  TITLE     Degradation of the peroxisome proliferator-activated receptor gamma 
            is linked to ligand-dependent activation.
  JOURNAL   J Biol Chem. 2000 Jun 16. 275(24):18527-33.
  PUBMED    10748014
REFERENCE   2  (bases 1 to 7042)
  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: "J Biol 
            Chem. 2000 Jun 16. 275(24):18527-33."
COMMENT     Sequence Label: pcDNA flag PPAR gamma
FEATURES             Location/Qualifiers
     source          1..7042
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     865..884
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        865..883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             988..1011
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2548..2577
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2593..2610
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2633..2650)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2639..2863
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2909..3337
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2996..3015)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3206..3227
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3346..3366)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3351..3680
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3531..3666
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3593..3612
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3747..4541
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3801..3820)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4411..4430
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4717..4838
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4754..4773)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4808..4827
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4887..4903)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4887..4903)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4900..4922)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4911..4927
                     /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(4935..4941,4942..4959,4960..4965))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4980..5001
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5118..5135)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5289..5877)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5369..5388)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6048..6839,6840..6908))
                     /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     6671..6690
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6909..7013)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta
      901 gttaagcttg gtaccgagct cggatccact agtccagtgt ggtggaattc tgcagatatc
      961 cagcacagtg gcggccgcac caccatggac tacaaagacg atgacgataa agcaatgggt
     1021 gaaactctgg gagattctcc tgttgaccca gagcatggtg ccttcgctga tgcactgcct
     1081 atgagcactt cacaagaaat taccatggtt gacacagaga tgccattctg gcccaccaac
     1141 ttcggaatca gctctgtgga cctctccgtg atggaagacc actcgcattc ctttgacatc
     1201 aagcccttta ccacagttga tttctccagc atttctgctc cacactatga agacattcca
     1261 ttcacaagag ctgacccaat ggttgctgat tacaaatatg acctgaagct ccaagaatac
     1321 caaagtgcga tcaaagtaga acctgcatct ccaccttatt attctgaaaa gacccagctc
     1381 tacaacaggc ctcatgaaga accttctaac tccctcatgg ccattgagtg ccgagtctgt
     1441 ggggataaag catcaggctt ccactatgga gttcatgctt gtgaaggatg caagggtttt
     1501 ttccgaagaa ccatccgatt gaagcttatt tatgataggt gtgatcttaa ctgccggatc
     1561 cacaaaaaaa gtagaaataa atgtcagtac tgtcggtttc agaagtgcct tgctgtgggg
     1621 atgtctcaca atgccatcag gtttgggcgg atgccacagg ccgagaagga gaagctgttg
     1681 gcggagatct ccagtgatat cgaccagctg aacccagagt ctgctgatct gcgagccctg
     1741 gcaaagcatt tgtatgactc atacataaag tccttcccgc tgaccaaagc caaggcgagg
     1801 gcgatcttga caggaaagac aacggacaaa tcaccatttg tcatctacga catgaattcc
     1861 ttaatgatgg gagaagataa aatcaagttc aaacatatca cccccctgca ggagcagagc
     1921 aaagaggtgg ccatccgaat ttttcaaggg tgccagtttc gatccgtaga agccgtgcaa
     1981 gagatcacag agtatgccaa aaatatccct ggtttcatta accttgattt gaatgaccaa
     2041 gtgactctgc tcaagtatgg tgtccatgag atcatctaca cgatgctggc ctccctgatg
     2101 aataaagatg gagtcctcat ctcagagggc caaggattca tgaccaggga gttcctcaaa
     2161 agcctgcgga agccctttgg tgactttatg gagcctaagt ttgagtttgc tgtgaagttc
     2221 aatgcactgg aattagatga cagtgacttg gctatattta tagctgtcat tattctcagt
     2281 ggagaccgcc caggcttgct gaacgtgaag cccatcgagg acatccaaga caacctgctg
     2341 caggccctgg aactgcagct caagctgaat cacccagagt cctctcagct gttcgccaag
     2401 gtgctccaga agatgacaga cctcaggcag atcgtcacag agcacgtgca gctactgcat
     2461 gtgatcaaga agacagagac agacatgagc cttcaccccc tgctccagga gatctacaag
     2521 gacttgtatt agtctagagg gcccttcgaa caaaaactca tctcagaaga ggatctgaat
     2581 atgcataccg gtcatcatca ccatcaccat tgagtttaaa cccgctgatc agcctcgact
     2641 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     2701 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     2761 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     2821 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     2881 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     2941 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     3001 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3061 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3121 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3181 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3241 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     3301 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     3361 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3421 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3481 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3541 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3601 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3661 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     3721 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     3781 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     3841 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3901 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     3961 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     4021 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     4081 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4141 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     4201 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     4261 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4321 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4381 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4441 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4501 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcgc
     4561 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     4621 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     4681 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     4741 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4801 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     4861 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     4921 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     4981 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     5041 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     5101 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     5161 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5221 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5281 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5341 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5401 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5461 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     5521 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5581 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5641 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5701 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     5761 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5821 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5881 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5941 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     6001 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     6061 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     6121 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6181 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6241 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6301 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6361 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6421 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6481 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6541 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6601 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6661 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6721 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6781 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6841 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6901 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6961 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     7021 ccccgaaaag tgccacctga cg
//
