LOCUS       sequence-1-expor        7203 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 7203)
  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 7203)
  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(delta CTD)
FEATURES             Location/Qualifiers
     source          1..7203
                     /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             2751..2768
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2791..2808)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2797..3021
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3067..3495
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3154..3173)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3364..3385
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3504..3524)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3509..3839
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3690..3825
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3752..3771
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3906..4700
                     /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(3960..3979)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4570..4589
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4876..4997
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4913..4932)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4967..4986
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5046..5062)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5046..5062)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5059..5081)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5070..5086
                     /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(5094..5100,5101..5118,5119..5124))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5139..5160
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5277..5294)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5448..6036)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5528..5547)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6207..6998,6999..7067))
                     /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     6830..6849
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7068..7172)
                     /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 aaaggatgct
     2701 cgagcatgca tctagttagt cagtctagac tagctgacta gaggaccggt catcatcacc
     2761 atcaccattg agtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
     2821 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     2881 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     2941 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     3001 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     3061 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     3121 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     3181 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg gggcatccct ttagggttcc
     3241 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     3301 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3361 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     3421 atttataagg gattttgggg atttcggcct attggttaaa aaatgagctg atttaacaaa
     3481 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     3541 ctccccaggc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     3601 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3661 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     3721 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     3781 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     3841 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     3901 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     3961 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4021 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4081 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4141 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4201 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4261 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4321 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4381 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     4441 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4501 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4561 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4621 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4681 ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg
     4741 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     4801 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     4861 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     4921 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     4981 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     5041 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     5101 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     5161 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5221 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     5281 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5341 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5401 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5461 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5521 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5581 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5641 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5701 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5761 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5821 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5881 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5941 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6001 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6061 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6121 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6181 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6241 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6301 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6361 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6421 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6481 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6541 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6601 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6661 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6721 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6781 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6841 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6901 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6961 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7021 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7081 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7141 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     7201 gtc
//
