LOCUS       sequence-2-expor        7905 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 7905)
  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 7905)
  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..7905
                     /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             974..997
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3453..3470
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3493..3510)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3499..3723
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3769..4197
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3856..3875)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4066..4087
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4206..4226)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4211..4541
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4392..4527
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4454..4473
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4608..5402
                     /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(4662..4681)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5272..5291
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5578..5699
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5615..5634)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5669..5688
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5748..5764)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5748..5764)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5761..5783)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5772..5788
                     /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(5796..5802,5803..5820,5821..5826))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5841..5862
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5979..5996)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6150..6738)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6230..6249)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6909..7700,7701..7769))
                     /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     7532..7551
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7770..7874)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gctggctagt
      901 taagcttggt accgagctcg gatccactag tccagtgtgg tggaattctg cagatatccg
      961 gatcaccacc atggactaca aagacgatga cgataaagca agaagagcat ctgtgggatc
     1021 ctgcagccaa gactctgtat ggagtgacat agagtgtgct gctctggttg gtgaggacca
     1081 gcctctttgc ccagatcttc ctgaacttga cctttctgaa cttgatgtga atgacttgga
     1141 tacagacagc tttctgggtg gattgaagtg gtgtagcgac caatcggaaa tcatatccaa
     1201 ccagtacaac aatgagcctg cgaacatatt tgagaagata gatgaagaga atgaggcaaa
     1261 cttgctagcg gtcctcacag agacactgga cagtctcccc gtggatgaag acggattgcc
     1321 ctcatttgat gcactgacag atggagccgt gaccactgac aacgaggcca gtccttcctc
     1381 catgcctgac ggcacccctc cccctcagga ggcagaagag ccgtctctac ttaagaagct
     1441 cttactggca ccagccaaca ctcagctcag ctacaatgaa tgcagcggtc ttagcactca
     1501 gaaccatgca gcaaaccaca cccacaggat cagaacaaac cctgccattg ttaagaccga
     1561 gaattcatgg agcaataaag cgaagagcat ttgtcaacag caaaagccac aaagacgtcc
     1621 ctgctcagag cttctcaagt atctgaccac aaacgatgac cctcctcaca ccaaacccac
     1681 agaaaacagg aacagcagca gagacaaatg tgcttccaaa aagaagtccc atacacaacc
     1741 gcagtcgcaa catgctcaag ccaaaccaac aactttatct cttcctctga ccccagagtc
     1801 accaaatgac cccaagggtt ccccatttga gaacaagact attgagcgaa ccttaagtgt
     1861 ggaactctct ggaactgcag gcctaactcc tcccacaact cctcctcata aagccaacca
     1921 agataaccct ttcaaggctt cgccaaagct gaagccctct tgcaagaccg tggtgccacc
     1981 gccaaccaag agggcccggt acagtgagtg ttctggtacc caaggcagcc actccaccaa
     2041 gaaagggccc gagcaatctg agttgtacgc acaactcagc aagtcctcag ggctcagccg
     2101 aggacacgag gaaaggaaga ctaaacggcc cagtctccgg ctgtttggtg accatgacta
     2161 ctgtcagtca ctcaattcca aaacggatat actcattaac atatcacagg agctccaaga
     2221 ctctagacaa ctagacttca aagatgcctc ctgtgactgg caggggcaca tctgttcttc
     2281 cacagattca ggccagtgct acctgagaga gactttggag gccagcaagc aggtctctcc
     2341 ttgcagcacc agaaaacagc tccaagacca ggaaatccga gcggagctga acaagcactt
     2401 cggtcatccc tgtcaagctg tgtttgacga caaatcagac aagaccagtg aactaaggga
     2461 tggcgacttc agtaatgaac aattctccaa actacctgtg tttataaatt caggactagc
     2521 catggatggc ctatttgatg acagtgaaga tgaaagtgat aaactgagct acccttggga
     2581 tggcacgcag ccctattcat tgttcgatgt gtcgccttct tgctcttcct ttaactctcc
     2641 gtgtcgagac tcagtgtcac caccgaaatc cttattttct caaagacccc aaaggatgcg
     2701 ctctcgttca agatcctttt ctcgacacag gtcgtgttcc cgatcaccat attccaggtc
     2761 aagatcaagg tccccaggca gtagatcctc ttcaagatcc tgttactact atgaatcaag
     2821 ccactacaga caccgcacac accgcaattc tcccttgtat gtgagatcac gttcaaggtc
     2881 accctacagc cgtaggccca ggtacgacag ctatgaagcc tatgagcacg aaaggctcaa
     2941 gagggatgaa taccgcaaag agcacgagaa gcgggagtct gaaagggcca aacagagaga
     3001 gaggcagaag cagaaagcaa ttgaagagcg ccgtgtgatt tacgttggta aaatcagacc
     3061 tgacacaacg cggacagaat tgagagaccg ctttgaagtt tttggtgaaa ttgaggaatg
     3121 caccgtaaat ctgcgggatg atggagacag ctatggtttc atcacctacc gttacacctg
     3181 tgacgctttc gctgctcttg agaatggata tactttacgc aggtcgaacg aaactgactt
     3241 cgagctgtac ttttgtggac ggaagcaatt tttcaagtct aactatgcag acctagatac
     3301 caactcagac gattttgacc ctgcttccac caagagcaag tatgactctc tggattttga
     3361 tagtttactg aaggaagctc agagaagctt gcgcaggtaa ctcgagcatg catctagtta
     3421 gtcagtctag actagctgac tagaggaccg gtcatcatca ccatcaccat tgagtttaaa
     3481 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     3541 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     3601 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     3661 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     3721 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     3781 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3841 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3901 ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc
     3961 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     4021 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4081 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg
     4141 ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     4201 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag gcaggcagaa
     4261 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     4321 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     4381 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     4441 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     4501 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     4561 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     4621 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     4681 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     4741 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     4801 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     4861 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     4921 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     4981 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     5041 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     5101 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     5161 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     5221 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     5281 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     5341 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     5401 gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     5461 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     5521 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     5581 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     5641 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     5701 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     5761 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     5821 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     5881 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     5941 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     6001 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     6061 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     6121 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     6181 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     6241 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     6301 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg
     6361 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     6421 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6481 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6541 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     6601 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6661 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6721 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6781 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6841 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6901 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6961 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     7021 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     7081 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     7141 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     7201 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     7261 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     7321 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     7381 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     7441 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     7501 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     7561 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     7621 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     7681 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     7741 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     7801 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     7861 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
