LOCUS       sequence-6781-ex        7404 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 7404)
  AUTHORS   Handschin C, Rhee J, Lin J, Tarr PT, Spiegelman BM
  TITLE     An autoregulatory loop controls peroxisome proliferator-activated 
            receptor gamma coactivator 1alpha expression in muscle.
  JOURNAL   Proc Natl Acad Sci U S A. 2003 Jun 10. 100(12):7111-6.
  PUBMED    12764228
REFERENCE   2  (bases 1 to 7404)
  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: "Proc Natl 
            Acad Sci U S A. 2003 Jun 10. 100(12):7111-6."
COMMENT     Sequence Label: PGC-1 alpha promoter luciferase delta MEF
FEATURES             Location/Qualifiers
     source          1..7404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      2668..2677
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2674..4326
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(2740..2758)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    4367..4488
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4378..4397)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4432..4451
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4736..4753)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4907..5495)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4987..5006)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5666..6457,6458..6526))
                     /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     6289..6308
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6527..6631)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6658..7113
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6745..6764)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6955..6976
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    7244..7292
                     /note="synthetic polyadenylation signal"
     misc_feature    7306..7397
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     7346..7365
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtacccctg tgctctctct agcttcacat accccttttt atgaaggtag aggacagagt
       61 ggctgtttta agcagatcat tagcttcatg gatgtgctgg gttagtttct tttcttttct
      121 tttcttttct ttttttaaag tagaattagg tggcaaaaaa agaaagaaag aaagaaagaa
      181 agaaagaaat tatcttttca aagcaaagaa aaagaaaatc ctgccacaat tcagtgtgag
      241 caagttaaga tatcaaaaca ataatgcaga gttatctagt gcaagcaacc atctgtaaga
      301 agaggtgtgc atgaggtttg aaaaagtatg gtggatgcct aatggatgac aaagagaaac
      361 caaagtatca gttaccatca ggatgccagg attgcttgat aatgatgaat aaacatcgac
      421 tctttatagc taagtggcca gcattatttc acagttgact gtgtggaaag tagagcccat
      481 gacctttgtc ctgaatttta atagtttact gaagttttac attaagtaaa aatctaattg
      541 gcaagaaaaa ttagtgtttc tttgtgggag tacagagtaa ggctactaat tgcatgtaga
      601 aaatgatccc agggttgtct ctctctctct ctgaattaga aaattaaatg aataatacct
      661 ttctgatgtt tggaagagga cagttgtagc agtgaagtat attcacctaa tttagaatgt
      721 caaagctgtt ggccatcttg ttctgttcac ttattgacaa actgtcgatt tcattaccat
      781 attccctgtc tactagttgt ggacatccaa gcaaacagac cccttatacg gggtctgaag
      841 caaacagcaa gcttgtgggc tttgtgtgtg tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg
      901 tgtgtgtgtg tgcacgctcg ctgcatttct ttctttcact ttactgtatt agtaactgac
      961 tatataaagt cagactgaga gaagtcacca atgttttcct tctaagggga gagaaaataa
     1021 agagcacatt aaattaacct cagtggaata ggagtttaaa tgaatggtgc tttataaatt
     1081 ccgcggagat gcatagggac ttttttcttc ttctaaatta ctgtcaagta aaggaaatga
     1141 gctggttttt gagtctgtgg ggtttgaggt attgtgaaaa tatgttttca aaggtcccct
     1201 gtgcatttct cactgggcct ggaagggtta agtctgagca cccaagtgtt atggaaagtg
     1261 ctgagagttg gttatgtcct ctgtctgtaa tgtcacagga aaaacagtgg cacctgcatt
     1321 acccctcatt gactcaggaa cgacaaaaaa gtattagtaa gcaaagctca agaaatgagt
     1381 atctctgctg ataccatttc agtgtttttc cttcattccc tggacattct tgatttcaaa
     1441 aacaaactgt acagcccaag gcactagggt tggagtccaa tgtttattca aaaaggcacc
     1501 ctgaagccat gaggaagact gtgctacata tgagaaaaga aataaggggt gggggcaggt
     1561 gagtagctaa gctgtttcag ggatggcagc agcaattgta ttttctagca tttgttttct
     1621 gggagcctat gagatccacg gaaagaatca tgagggggaa cccaagagtc tagggtgttg
     1681 tggcttgctt gctttacaag gagcaaggca aactgcagta acagtttagg agactgcatt
     1741 ctctactgcc aaggagacag ctgatttggg gtagagaaat ttgtttagac ctaaacaaat
     1801 gtggcggttt tgttgactaa acatggaaag aaagaaagaa agaaagaaag aaagaaagaa
     1861 agaaagaaag aaagaaagaa agaaacaaag aaagaaagaa agaaaggaag gaaggaagga
     1921 agaaaggaag aaaggaagga aggaaggaag gaaagaaagg agagagagaa agaaaatcgg
     1981 gggtgttgcc ttcaaacact cctctaatag ggagggaaaa aaaagaatct catgaaaatg
     2041 tatcacatga ggagcgcttg cttcagttcc aagctgagtc tggggctact tggaaaccat
     2101 ttcttaaagc acacacattt taggcaaggg tgtagttact gtgtcagtaa caggggatct
     2161 ttgctatttg cctgttttgg atggaaaata aatttaaaaa aaaaagattg caggagattt
     2221 gagttattat gtgagcaggg ctccggttta gagttggtgg cattcaaagc tggcttcagt
     2281 cacagtgtga tgcttgaagc ctcccaaagg ccaagtgttt ccttttcttt cttctatttt
     2341 ttttttcctc tctctctaag cgttacttca ctgaggcaga gggctgcctt ggagtgacgt
     2401 caggagtttg tgcagcaagc ttgcacagga gaagggaggc tgggtgagtg acagcccagc
     2461 ctacttttta atagctttgt catgtgactg gggactgtag taagacaggt gccttcagtt
     2521 cactctcagt aaggggctgg ttgcctgcat gagtgtgtgc tgtgtgtcag agtggattgg
     2581 agttgaaaaa gcttgactgg cgtcattcgg gagctggctc gagatctgcg atctaagtaa
     2641 gcttggcatt ccggtactgt tggtaaagcc accatggaag acgccaaaaa cataaagaaa
     2701 ggcccggcgc cattctatcc gctggaagat ggaaccgctg gagagcaact gcataaggct
     2761 atgaagagat acgccctggt tcctggaaca attgctttta cagatgcaca tatcgaggtg
     2821 gacatcactt acgctgagta cttcgaaatg tccgttcggt tggcagaagc tatgaaacga
     2881 tatgggctga atacaaatca cagaatcgtc gtatgcagtg aaaactctct tcaattcttt
     2941 atgccggtgt tgggcgcgtt atttatcgga gttgcagttg cgcccgcgaa cgacatttat
     3001 aatgaacgtg aattgctcaa cagtatgggc atttcgcagc ctaccgtggt gttcgtttcc
     3061 aaaaaggggt tgcaaaaaat tttgaacgtg caaaaaaagc tcccaatcat ccaaaaaatt
     3121 attatcatgg attctaaaac ggattaccag ggatttcagt cgatgtacac gttcgtcaca
     3181 tctcatctac ctcccggttt taatgaatac gattttgtgc cagagtcctt cgatagggac
     3241 aagacaattg cactgatcat gaactcctct ggatctactg gtctgcctaa aggtgtcgct
     3301 ctgcctcata gaactgcctg cgtgagattc tcgcatgcca gagatcctat ttttggcaat
     3361 caaatcattc cggatactgc gattttaagt gttgttccat tccatcacgg ttttggaatg
     3421 tttactacac tcggatattt gatatgtgga tttcgagtcg tcttaatgta tagatttgaa
     3481 gaagagctgt ttctgaggag ccttcaggat tacaagattc aaagtgcgct gctggtgcca
     3541 accctattct ccttcttcgc caaaagcact ctgattgaca aatacgattt atctaattta
     3601 cacgaaattg cttctggtgg cgctcccctc tctaaggaag tcggggaagc ggttgccaag
     3661 aggttccatc tgccaggtat caggcaagga tatgggctca ctgagactac atcagctatt
     3721 ctgattacac ccgaggggga tgataaaccg ggcgcggtcg gtaaagttgt tccatttttt
     3781 gaagcgaagg ttgtggatct ggataccggg aaaacgctgg gcgttaatca aagaggcgaa
     3841 ctgtgtgtga gaggtcctat gattatgtcc ggttatgtaa acaatccgga agcgaccaac
     3901 gccttgattg acaaggatgg atggctacat tctggagaca tagcttactg ggacgaagac
     3961 gaacacttct tcatcgttga ccgcctgaag tctctgatta agtacaaagg ctatcaggtg
     4021 gctcccgctg aattggaatc catcttgctc caacacccca acatcttcga cgcaggtgtc
     4081 gcaggtcttc ccgacgatga cgccggtgaa cttcccgccg ccgttgttgt tttggagcac
     4141 ggaaagacga tgacggaaaa agagatcgtg gattacgtcg ccagtcaagt aacaaccgcg
     4201 aaaaagttgc gcggaggagt tgtgtttgtg gacgaagtac cgaaaggtct taccggaaaa
     4261 ctcgacgcaa gaaaaatcag agagatcctc ataaaggcca agaagggcgg aaagatcgcc
     4321 gtgtaattct agagtcgggg cggccggccg cttcgagcag acatgataag atacattgat
     4381 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     4441 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     4501 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
     4561 aacctctaca aatgtggtaa aatcgataag gatccgtcga ccgatgccct tgagagcctt
     4621 caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac
     4681 tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctcttcc gcttcctcgc
     4741 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     4801 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4861 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4921 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4981 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5041 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5101 aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5161 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5221 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5281 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5341 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5401 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5461 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     5521 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5581 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5641 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5701 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5761 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5821 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5881 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5941 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6001 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6061 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6121 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6181 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6241 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6301 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6361 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6421 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6481 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     6541 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6601 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     6661 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     6721 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     6781 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     6841 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     6901 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     6961 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     7021 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     7081 cgaattttaa caaaatatta acgtttacaa tttcccattc gccattcagg ctgcgcaact
     7141 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagcccaag ctaccatgat
     7201 aagtaagtaa tattaaggta cgggaggtac ttggagcggc cgcaataaaa tatctttatt
     7261 ttcattacat ctgtgtgttg gttttttgtg tgaatcgata gtactaacat acgctctcca
     7321 tcaaaacaaa acgaaacaaa acaaactagc aaaataggct gtccccagtg caagtgcagg
     7381 tgccagaaca tttctctatc gata
//
