LOCUS       sequence-429987-        7838 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 7838)
  AUTHORS   Ruas JL, White JP, Rao RR, Kleiner S, Brannan KT, Harrison BC, 
            Greene NP, Wu J, Estall JL, Irving BA, Lanza IR, Rasbach KA, Okutsu 
            M, Nair KS, Yan Z, Leinwand LA, Spiegelman BM
  TITLE     A PGC-1alpha isoform induced by resistance training regulates 
            skeletal muscle hypertrophy.
  JOURNAL   Cell. 2012 Dec 7;151(6):1319-31. doi: 10.1016/j.cell.2012.10.050.
  PUBMED    23217713
REFERENCE   2  (bases 1 to 7838)
  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: 
            "10.1016/j.cell.2012.10"; journalName: "Cell"; date: "2012-12-7- 7";
            volume: "151"; issue: "6"; pages: "1319-31"
COMMENT     Sequence Label: pcDNA3.1-Flag-PGC-1alpha1
FEATURES             Location/Qualifiers
     source          1..7838
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             919..942
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3343..3360
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(3391..3409)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3392..3409)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3429..3446)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3435..3659
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3705..4133
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3792..3811)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4002..4023
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4142..4162)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4147..4476
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4327..4462
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4389..4408
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4543..5337
                     /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(4597..4616)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5207..5226
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5511..5632
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5548..5567)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5602..5621
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5681..5697)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5681..5697)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5694..5716)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5705..5721
                     /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(5729..5735,5736..5753,5754..5759))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5774..5795
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5912..5929)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6083..6671)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6163..6182)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6842..7633,7634..7702))
                     /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     7465..7484
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7703..7807)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccaccatgga ctacaaagac gatgacgaca aggcggccgc gatggcttgg
      961 gacatgtgca gccaagactc tgtatggagt gacatagagt gtgctgctct ggttggtgag
     1021 gaccagcctc tttgcccaga tcttcctgaa cttgaccttt ctgaacttga tgtgaatgac
     1081 ttggatacag acagctttct gggtggattg aagtggtgta gcgaccaatc ggaaatcata
     1141 tccaaccagt acaacaatga gcctgcgaac atatttgaga agatagatga agagaatgag
     1201 gcaaacttgc tagcggtcct cacagagaca ctggacagtc tccccgtgga tgaagacgga
     1261 ttgccctcat ttgatgcact gacagatgga gccgtgacca ctgacaacga ggccagtcct
     1321 tcctccatgc ctgacggcac ccctccccct caggaggcag aagagccgtc tctacttaag
     1381 aagctcttac tggcaccagc caacactcag ctcagctaca atgaatgcag cggtcttagc
     1441 actcagaacc atgcagcaaa ccacacccac aggatcagaa caaaccctgc cattgttaag
     1501 accgagaatt catggagcaa taaagcgaag agcatttgtc aacagcaaaa gccacaaaga
     1561 cgtccctgct cagagcttct caagtatctg accacaaacg atgaccctcc tcacaccaaa
     1621 cccacagaaa acaggaacag cagcagagac aaatgtgctt ccaaaaagaa gtcccataca
     1681 caaccgcagt cgcaacatgc tcaagccaaa ccaacaactt tatctcttcc tctgacccca
     1741 gagtcaccaa atgaccccaa gggttcccca tttgaaaaca agactattga gcgaacctta
     1801 agtgtggaac tctctggaac tgcaggccta actcctccca caactcctcc tcataaagcc
     1861 aaccaagata accctttcaa ggcttcgcca aagctgaagc cctcttgcaa gaccgtggtg
     1921 ccaccgccaa ccaagagggc ccggtacagt gagtgttctg gtaccaaagg cagccactcc
     1981 accaagaaag ggcccgagca atctgagttg tacgcacaac tcagcaagtc ctcagggctc
     2041 agccgaggac acgaggaaag gaagactaaa cggcccagtc tccggctgtt tggtgaccat
     2101 gactactgtc agtcactcaa ttccaaaacg gatatactca ttaacatatc acaggagctc
     2161 caagactcta gacaactaga cttcaaagat gcctcctgtg actggcaggg gcacatctgt
     2221 tcttccacag attcaggcca gtgctacctg agagagactt tggaggccag caagcaggtc
     2281 tctccttgca gcaccagaaa acagctccaa gaccaggaaa tccgagcgga gctgaacaag
     2341 cacttcggtc atccctgtca agctgtgttt gacgacaaat cagacaagac cagtgaacta
     2401 agggatggcg acttcagtaa tgaacaattc tccaaactac ctgtgtttat aaattcagga
     2461 ctagccatgg atggcctatt tgatgacagt gaagatgaaa gtgataaact gagctaccct
     2521 tgggatggca cgcagcccta ttcattgttc gatgtgtcgc cttcttgctc ttcctttaac
     2581 tctccgtgtc gagactcagt gtcaccaccg aaatccttat tttctcaaag accccaaagg
     2641 atgcgctctc gttcaagatc cttttctcga cacaggtcgt gttcccgatc accatattcc
     2701 aggtcaagat caaggtcccc aggcagtaga tcctcttcaa gatcctgtta ctactatgaa
     2761 tcaagccact acagacaccg cacacaccgc aattctccct tgtatgtgag atcacgttca
     2821 aggtcaccct acagccgtag gcccaggtac gacagctatg aagcctatga gcacgaaagg
     2881 ctcaagaggg atgaataccg caaagagcac gagaagcggg agtctgaaag ggccaaacag
     2941 agagagaggc agaagcagaa agcaattgaa gagcgccgtg tgatttacgt tggtaaaatc
     3001 agacctgaca caacgcggac agaattgaga gaccgctttg aagtttttgg tgaaattgag
     3061 gaatgcaccg taaatctgcg ggatgatgga gacagctatg gtttcatcac ctaccgttac
     3121 acctgtgacg ctttcgctgc tcttgagaat ggatatactt tacgcaggtc gaacgaaact
     3181 gacttcgagc tgtacttttg tggacggaag caatttttca agtctaacta tgcagaccta
     3241 gataccaact cagacgattt tgaccctgct tccaccaaga gcaagtatga ctctctggat
     3301 tttgatagtt tactgaagga agctcagaga agcttgcgca ggcatcatca tcatcatcat
     3361 taactcgagc atgcatctag agggccctat tctatagtgt cacctaaatg ctagagctcg
     3421 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     3481 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     3541 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     3601 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     3661 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     3721 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3781 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3841 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     3901 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3961 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4021 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     4081 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     4141 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4201 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4261 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4321 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4381 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     4441 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     4501 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4561 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4621 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4681 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     4741 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4801 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4861 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4921 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4981 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5041 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     5101 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5161 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5221 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5281 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5341 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     5401 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5461 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     5521 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5581 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     5641 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     5701 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     5761 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     5821 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     5881 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5941 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     6001 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6061 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6121 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6181 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6241 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6301 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6361 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6421 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6481 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6541 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6601 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6661 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6721 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6781 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6841 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6901 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6961 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     7021 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     7081 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7141 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7201 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     7261 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     7321 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     7381 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     7441 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7501 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7561 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7621 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7681 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7741 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7801 ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
