LOCUS       sequence-258338-        7338 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 7338)
  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 7338)
  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 2kb luciferase
FEATURES             Location/Qualifiers
     source          1..7338
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      2598..2607
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2604..4256
                     /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(2670..2688)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    4297..4418
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4308..4327)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4362..4381
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4666..4683)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4837..5425)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4917..4936)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5596..6387,6388..6456))
                     /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     6219..6238
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6457..6561)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6588..7043
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6675..6694)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6885..6906
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    7174..7222
                     /note="synthetic polyadenylation signal"
     misc_feature    7236..7327
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     7276..7295
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 cctgtgctct ctctagcttc acatacccct ttttatgaag gtagaggaca gagtggctgt
       61 tttaagcaga tcattagctt catggatgtg ctgggttagt ttcttttctt ttcttttctt
      121 ttttttttta aagtagaatt aggtggcaaa aaaaaaaaag aaagaaagaa agaaagaaag
      181 aaagaaagaa agaaagaaag aaattatctt ttcaaagcaa agagaaagaa aatcctgcca
      241 taattcagtg tgagcaagtt aagatatcaa aacaataatg cagagttatc tagtgcaagc
      301 aaccatctgt aagaagaggt gcgcatgagg tttgaaaaag tatggtggat gcctaatgga
      361 tgacaaagag aaaccaaagt atcagttacc atcaggatgc caggattgct tgataatgat
      421 gaataaacat cgactcttta tagctaagtg gccagcatta tttcacagtt gactgtgtgg
      481 aaagtagagc ccatgacctt tgtcctgaat tttaatagtt tactgaagtt ttacattaag
      541 taaaaatcta attggcaaga aaaattagtg tttctttgtg ggagtacaga gtaaggctac
      601 taattgcatg tagaaaatga tcccagggtt gtctctctct ctctctgaat tagaaaatta
      661 aatgaataat acctttctga tgtttggaag aggacagttg tagcagtgaa gtatattcac
      721 ctaatttaga atgtcaaagc tgttggccat cttgttctgt tcacttattg acaaactgtc
      781 gatttcatta ccatattccc tgtctactag ttgtggacat ccaagcaaac agacccctta
      841 tacggggtct gaagcaaaca gcaagcttgt gggctttgtg tgtgtgtgtg tgtgtgtgtg
      901 tgtgtgcacg ctcgctgcat ttctttcttt cactttactg tattagtaac tgactatata
      961 aagtcagact gagagaagtc accaatgttt tccttctaag gggagagaaa ataaagagca
     1021 cattaaatta acctcagtgg aataggagtt taaatgaatg gtgctttata aattatattt
     1081 agatgcatag ggactttttt cttcttctaa attactgtca agtaaaggaa atgagctggt
     1141 ttttgagtct gtggggtttg aggtattgtg aaaatatgtt ttcaaaggtg catttctcac
     1201 tgggcctgga agggttaagt ctgagcaccc aagtgttatg gaaagtgctg agagttggtt
     1261 atgtcctctg tctgtaatgt cacaggaaaa acagtggcac ctgcattacc cctcattgac
     1321 tcaggaatga caaaaaagta ttagtaagca aagctcaaga aatgagtatc tctgctgata
     1381 ccatttcagt gtttttcctt cattccctgg acattcttga tttcaaaaac aaactgtaca
     1441 gcccaaggca ctagggttgg agtccaatgt ttattcaaaa aggcaccctg aagccatgag
     1501 gaagactgtg ctacatatga gaaaagaaat aaggggtggg ggcaggtgag tagctaagct
     1561 gtttcaggga tggcagcagc aattgtattt tctagcattt gttttctggg agcctatgag
     1621 atccacggaa agaatcatga gggggaaccc aagagtctag ggtgttgtgg cttgcttgct
     1681 ttactaggag caaggcaaac tgcagtaaca gtttaggaga ctgcattctc tactgccaag
     1741 gagacagctg atttggggta gagaaatttg tttagaccta aacaaatgtg gcggttttgt
     1801 tgactaaaca tggaaagaaa gaaagaaaga aagaaagaaa gaaacaaaga aagaaagaaa
     1861 gaaagaaagg aaggaaggaa ggaaagaaag gagagagaga aagaaaatcg ggggtgttgc
     1921 cttcaaacac tcctctaata gggagggaaa aaaaagaatc tcatgaaaat gtatcacatg
     1981 aggagcgctt gcttcagttc caagctgagt ctggggctac ttggaaacca tttcttaaag
     2041 cacacacatt ttaggcaagg gtgtagttac tgtgtcagta acaggggatc tttgctattt
     2101 gcctgttttg gatggaaaat aaattaaaaa aaaaaagatt gcaggagatt tgagttatta
     2161 tgtgagcagg gctccggttt agagttggtg gcattcaaag ctggcttcag tcacagtgtg
     2221 atgcttgaag cctcccaaag gccaagtgtt tccttttctt tcttctattt ttttttcctc
     2281 tctctctaag cgttacttca ctgaggcaga gggctgcctc ggagtgacgt caggagtttg
     2341 tgcagcaagc ttgcacagga gaagggaggc tgggtgagtg acagcccagc ctacttttta
     2401 atagctttgt catgtgactg gggactgtag taagacaggt gccttcagtt cactctcagt
     2461 aaggggctgg ttgcctgcat gagtgtgtgc tgtgtgtcag agtggattgg agttgaaaaa
     2521 gcttgactgg cgtcattcgg gagctggctc gagatctgcg atctaagtaa gcttggcatt
     2581 ccggtactgt tggtaaagcc accatggaag acgccaaaaa cataaagaaa ggcccggcgc
     2641 cattctatcc gctggaagat ggaaccgctg gagagcaact gcataaggct atgaagagat
     2701 acgccctggt tcctggaaca attgctttta cagatgcaca tatcgaggtg gacatcactt
     2761 acgctgagta cttcgaaatg tccgttcggt tggcagaagc tatgaaacga tatgggctga
     2821 atacaaatca cagaatcgtc gtatgcagtg aaaactctct tcaattcttt atgccggtgt
     2881 tgggcgcgtt atttatcgga gttgcagttg cgcccgcgaa cgacatttat aatgaacgtg
     2941 aattgctcaa cagtatgggc atttcgcagc ctaccgtggt gttcgtttcc aaaaaggggt
     3001 tgcaaaaaat tttgaacgtg caaaaaaagc tcccaatcat ccaaaaaatt attatcatgg
     3061 attctaaaac ggattaccag ggatttcagt cgatgtacac gttcgtcaca tctcatctac
     3121 ctcccggttt taatgaatac gattttgtgc cagagtcctt cgatagggac aagacaattg
     3181 cactgatcat gaactcctct ggatctactg gtctgcctaa aggtgtcgct ctgcctcata
     3241 gaactgcctg cgtgagattc tcgcatgcca gagatcctat ttttggcaat caaatcattc
     3301 cggatactgc gattttaagt gttgttccat tccatcacgg ttttggaatg tttactacac
     3361 tcggatattt gatatgtgga tttcgagtcg tcttaatgta tagatttgaa gaagagctgt
     3421 ttctgaggag ccttcaggat tacaagattc aaagtgcgct gctggtgcca accctattct
     3481 ccttcttcgc caaaagcact ctgattgaca aatacgattt atctaattta cacgaaattg
     3541 cttctggtgg cgctcccctc tctaaggaag tcggggaagc ggttgccaag aggttccatc
     3601 tgccaggtat caggcaagga tatgggctca ctgagactac atcagctatt ctgattacac
     3661 ccgaggggga tgataaaccg ggcgcggtcg gtaaagttgt tccatttttt gaagcgaagg
     3721 ttgtggatct ggataccggg aaaacgctgg gcgttaatca aagaggcgaa ctgtgtgtga
     3781 gaggtcctat gattatgtcc ggttatgtaa acaatccgga agcgaccaac gccttgattg
     3841 acaaggatgg atggctacat tctggagaca tagcttactg ggacgaagac gaacacttct
     3901 tcatcgttga ccgcctgaag tctctgatta agtacaaagg ctatcaggtg gctcccgctg
     3961 aattggaatc catcttgctc caacacccca acatcttcga cgcaggtgtc gcaggtcttc
     4021 ccgacgatga cgccggtgaa cttcccgccg ccgttgttgt tttggagcac ggaaagacga
     4081 tgacggaaaa agagatcgtg gattacgtcg ccagtcaagt aacaaccgcg aaaaagttgc
     4141 gcggaggagt tgtgtttgtg gacgaagtac cgaaaggtct taccggaaaa ctcgacgcaa
     4201 gaaaaatcag agagatcctc ataaaggcca agaagggcgg aaagatcgcc gtgtaattct
     4261 agagtcgggg cggccggccg cttcgagcag acatgataag atacattgat gagtttggac
     4321 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     4381 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     4441 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
     4501 aatgtggtaa aatcgataag gatccgtcga ccgatgccct tgagagcctt caacccagtc
     4561 agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac tgtcttcttt
     4621 atcatgcaac tcgtaggaca ggtgccggca gcgctcttcc gcttcctcgc tcactgactc
     4681 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4741 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4801 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4861 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4921 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4981 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5041 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5101 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5161 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5221 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     5281 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5341 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5401 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5461 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5521 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5581 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5641 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5701 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5761 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5821 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5881 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5941 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6001 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6061 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6121 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6181 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6241 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6301 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6361 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6421 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6481 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6541 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag
     6601 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     6661 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     6721 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     6781 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     6841 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     6901 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     6961 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     7021 caaaatatta acgcttacaa tttgccattc gccattcagg ctgcgcaact gttgggaagg
     7081 gcgatcggtg cgggcctctt cgctattacg ccagcccaag ctaccatgat aagtaagtaa
     7141 tattaaggta cgggaggtac ttggagcggc cgcaataaaa tatctttatt ttcattacat
     7201 ctgtgtgttg gttttttgtg tgaatcgata gtactaacat acgctctcca tcaaaacaaa
     7261 acgaaacaaa acaaactagc aaaataggct gtccccagtg caagtgcagg tgccagaaca
     7321 tttctctatc gataggta
//
