LOCUS       sequence-220058-        7904 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 7904)
  AUTHORS   Monsalve M, Wu Z, Adelmant G, Puigserver P, Fan M, Spiegelman BM
  TITLE     Direct coupling of transcription and mRNA processing through the 
            thermogenic coactivator PGC-1.
  JOURNAL   Mol Cell 2000 Aug;6(2):307-16.
  PUBMED    10983978
REFERENCE   2  (bases 1 to 7904)
  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: "Mol Cell"; 
            date: "2000-08"; volume: "6(2)"; pages: "307-16"
COMMENT     Sequence Label: pcDNA-f:PGC1
FEATURES             Location/Qualifiers
     source          1..7904
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             740..763
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3219..3236
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3259..3276)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3265..3489
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3535..3963
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3622..3641)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3832..3853
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3972..3992)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3977..4306
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4157..4292
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4219..4238
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4373..5167
                     /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(4427..4446)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5037..5056
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5343..5464
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5380..5399)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5434..5453
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5513..5529)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5513..5529)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5526..5548)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5537..5553
                     /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(5561..5567,5568..5585,5586..5591))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5606..5627
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5744..5761)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5915..6503)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5995..6014)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6674..7465,7466..7534))
                     /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     7297..7316
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7535..7639)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7714..7733)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagttaag cttggtaccg agctcggatc cactagtcca gtgtggtgga attctgcaga
      721 tatccggatc accaccatgg actacaaaga cgatgacgat aaagcaagaa gagcatctgt
      781 gggatcctgc agccaagact ctgtatggag tgacatagag tgtgctgctc tggttggtga
      841 ggaccagcct ctttgcccag atcttcctga acttgacctt tctgaacttg atgtgaatga
      901 cttggataca gacagctttc tgggtggatt gaagtggtgt agcgaccaat cggaaatcat
      961 atccaaccag tacaacaatg agcctgcgaa catatttgag aagatagatg aagagaatga
     1021 ggcaaacttg ctagcggtcc tcacagagac actggacagt ctccccgtgg atgaagacgg
     1081 attgccctca tttgatgcac tgacagatgg agccgtgacc actgacaacg aggccagtcc
     1141 ttcctccatg cctgacggca cccctccccc tcaggaggca gaagagccgt ctctacttaa
     1201 gaagctctta ctggcaccag ccaacactca gctcagctac aatgaatgca gcggtcttag
     1261 cactcagaac catgcagcaa accacaccca caggatcaga acaaaccctg ccattgttaa
     1321 gaccgagaat tcatggagca ataaagcgaa gagcatttgt caacagcaaa agccacaaag
     1381 acgtccctgc tcagagcttc tcaagtatct gaccacaaac gatgaccctc ctcacaccaa
     1441 acccacagaa aacaggaaca gcagcagaga caaatgtgct tccaaaaaga agtcccatac
     1501 acaaccgcag tcgcaacatg ctcaagccaa accaacaact ttatctcttc ctctgacccc
     1561 agagtcacca aatgacccca agggttcccc atttgagaac aagactattg agcgaacctt
     1621 aagtgtggaa ctctctggaa ctgcaggcct aactcctccc acaactcctc ctcataaagc
     1681 caaccaagat aaccctttca aggcttcgcc aaagctgaag ccctcttgca agaccgtggt
     1741 gccaccgcca accaagaggg cccggtacag tgagtgttct ggtacccaag gcagccactc
     1801 caccaagaaa gggcccgagc aatctgagtt gtacgcacaa ctcagcaagt cctcagggct
     1861 cagccgagga cacgaggaaa ggaagactaa acggcccagt ctccggctgt ttggtgacca
     1921 tgactactgt cagtcactca attccaaaac ggatatactc attaacatat cacaggagct
     1981 ccaagactct agacaactag acttcaaaga tgcctcctgt gactggcagg ggcacatctg
     2041 ttcttccaca gattcaggcc agtgctacct gagagagact ttggaggcca gcaagcaggt
     2101 ctctccttgc agcaccagaa aacagctcca agaccaggaa atccgagcgg agctgaacaa
     2161 gcacttcggt catccctgtc aagctgtgtt tgacgacaaa tcagacaaga ccagtgaact
     2221 aagggatggc gacttcagta atgaacaatt ctccaaacta cctgtgttta taaattcagg
     2281 actagccatg gatggcctat ttgatgacag tgaagatgaa agtgataaac tgagctaccc
     2341 ttgggatggc acgcagccct attcattgtt cgatgtgtcg ccttcttgct cttcctttaa
     2401 ctctccgtgt cgagactcag tgtcaccacc gaaatcctta ttttctcaaa gaccccaaag
     2461 gatgcgctct cgttcaagat ccttttctcg acacaggtcg tgttcccgat caccatattc
     2521 caggtcaaga tcaaggtccc caggcagtag atcctcttca agatcctgtt actactatga
     2581 atcaagccac tacagacacc gcacacaccg caattctccc ttgtatgtga gatcacgttc
     2641 aaggtcaccc tacagccgta ggcccaggta cgacagctat gaagcctatg agcacgaaag
     2701 gctcaagagg gatgaatacc gcaaagagca cgagaagcgg gagtctgaaa gggccaaaca
     2761 gagagagagg cagaagcaga aagcaattga agagcgccgt gtgatttacg ttggtaaaat
     2821 cagacctgac acaacgcgga cagaattgag agaccgcttt gaagtttttg gtgaaattga
     2881 ggaatgcacc gtaaatctgc gggatgatgg agacagctat ggtttcatca cctaccgtta
     2941 cacctgtgac gctttcgctg ctcttgagaa tggatatact ttacgcaggt cgaacgaaac
     3001 tgacttcgag ctgtactttt gtggacggaa gcaatttttc aagtctaact atgcagacct
     3061 agataccaac tcagacgatt ttgaccctgc ttccaccaag agcaagtatg actctctgga
     3121 ttttgatagt ttactgaagg aagctcagag aagcttgcgc aggtaactcg agcatgcatc
     3181 tagttagtca gtctagacta gctgactaga ggaccggtca tcatcaccat caccattgag
     3241 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     3301 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     3361 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     3421 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     3481 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     3541 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3601 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3661 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3721 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     3781 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3841 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     3901 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3961 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     4021 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     4081 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     4141 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     4201 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     4261 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     4321 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     4381 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     4441 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     4501 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     4561 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     4621 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     4681 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     4741 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     4801 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     4861 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     4921 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     4981 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     5041 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     5101 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     5161 cttctgagcg ggactctggg gttcgcgaaa tgaccgacca agcgacgccc aacctgccat
     5221 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     5281 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5341 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     5401 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     5461 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     5521 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5581 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5641 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5701 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5761 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5821 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5881 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5941 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     6001 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     6061 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     6121 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6181 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6241 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6301 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     6361 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6421 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     6481 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6541 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6601 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6661 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6721 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6781 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6841 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6901 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6961 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     7021 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     7081 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     7141 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     7201 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     7261 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     7321 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     7381 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7441 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7501 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7561 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7621 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
     7681 gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     7741 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
     7801 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
     7861 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgtt
//
