LOCUS       sequence-197589-        5630 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5630)
  AUTHORS   Ichida M, Nemoto S, Finkel T
  TITLE     Identification of a specific molecular repressor of the peroxisome 
            proliferator-activated receptor gamma Coactivator-1 alpha 
            (PGC-1alpha).
  JOURNAL   J Biol Chem. 2002 Dec 27. 277(52):50991-5.
  PUBMED    12397057
REFERENCE   2  (bases 1 to 5630)
  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. 2002 Dec 27. 277(52):50991-5."
COMMENT     Sequence Label: pCMV flag ERR alpha
FEATURES             Location/Qualifiers
     source          1..5630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        67..370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        371..574
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     525..545
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        620..638
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      673..682
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             682..705
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2059..2075)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2060..2076)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(2131..2150)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2132..2150)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2424..2545
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2461..2480)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2515..2534
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2552..3007)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2689..2710)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2901..2920
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3034..3138
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3140..3497
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3140..3331
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3163..3183)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3348..3483
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3532..4326
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3532..4326
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3586..3605)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4196..4215
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4514..4533)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4558..4605
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4934..5522
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5423..5442
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
       61 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      121 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      181 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      241 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      301 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      361 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      421 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      481 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      541 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      601 gcgattacgc caagctcgaa attaaccctc actaaaggga acaaaagctg gagctccacc
      661 gcggtggcgg ccgccaccat ggattacaag gatgacgacg ataaggcccg ggcggatccc
      721 ccgggctgca ggaattcgcc cttcatgtcc agccaggtgg tgggcattga gcctctctac
      781 atcaaggcag agccggccag ccctgacagt ccaaagggtt cctcggagac agagaccgag
      841 cctcctgtgg ccctggcccc tggtccagct cccactcgct gcctcccagg ccacaaggaa
      901 gaggaggatg gggagggggc tgggcctggc gagcagggcg gtgggaagct ggtgctcagc
      961 tccctgccca agcgcctctg cctggtctgt ggggacgtgg cctccggcta ccactatggt
     1021 gtggcatcct gtgaggcctg caaagccttc ttcaagagga ccatccaggg gagcatcgag
     1081 tacagctgtc cggcctccaa cgagtgtgag atcaccaagc ggagacgcaa ggcctgccag
     1141 gcctgccgct tcaccaagtg cctgcgggtg ggcatgctca aggagggagt gcgcctggac
     1201 cgcgtccggg gtgggcggca gaagtacaag cggcggccgg aggtggaccc actgcccttc
     1261 ccgggcccct tccctgctgg gcccctggca gtcgctggag gcccccggaa gacagcagcc
     1321 ccagtgaatg cactggtgtc tcatctgctg gtggttgagc ctgagaagct ctatgccatg
     1381 cctgaccccg caggccctga tgggcacctc ccagccgtgg ctaccctctg tgacctcttt
     1441 gaccgagaga ttgtggtcac catcagctgg gccaagagca tcccaggctt ctcatcgctg
     1501 tcgctgtctg accagatgtc agtactgcag agcgtgtgga tggaggtgct ggtgctgggt
     1561 gtggcccagc gctcactgcc actgcaggat gagctggcct tcgctgagga cttagtcctg
     1621 gatgaagagg gggcacgggc agctggcctg ggggaactgg gggctgccct gctgcaacta
     1681 gtgcggcggc tgcaggccct gcggctggag cgagaggagt atgttctact aaaggccttg
     1741 gcccttgcca attcagactc tgtgcacatc gaagatgccg aggctgtgga gcagctgcga
     1801 gaagctctgc acgaggccct gctggagtat gaagccggcc gggctggccc cggagggggt
     1861 gctgagcggc ggcgggcggg caggctgctg ctcacgctac cgctcctccg ccagacagcg
     1921 ggcaaagtgc tggcccattt ctatggggtg aagctggagg gcaaggtgcc catgcacaag
     1981 ctgttcttgg agatgctcga ggccatgatg gactgaggca aggggtggga ctaagggcga
     2041 attcgatatc aagcttatcg ataccgtcga cctcgagggg gggcccggta ccttaattaa
     2101 ttaaggtacc aggtaagtgt acccaattcg ccctatagtg agtcgtatta caattcactc
     2161 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggagatccaa tttttaagtg
     2221 tataatgtgt taaactactg attctaattg tttgtgtatt ttagattcac agtcccaagg
     2281 ctcatttcag gcccctcagt cctcacagtc tgttcatgat cataatcagc cataccacat
     2341 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     2401 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     2461 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     2521 tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa
     2581 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     2641 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     2701 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     2761 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     2821 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     2881 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     2941 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     3001 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     3061 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     3121 tattgaaaaa ggaagaatcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     3181 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3241 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3301 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3361 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3421 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3481 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     3541 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3601 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3661 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     3721 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     3781 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     3841 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     3901 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     3961 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agaacatcag
     4021 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     4081 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4141 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     4201 gctacccgtg atattgctga agaacttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4261 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     4321 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     4381 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4441 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     4501 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     4561 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     4621 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     4681 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     4741 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     4801 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     4861 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     4921 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4981 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5041 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     5101 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5161 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5221 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5281 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5341 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     5401 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     5461 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     5521 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     5581 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
//
