LOCUS       sequence-473152-        6716 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient overexpression of FBXO15 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6716)
  AUTHORS   Donato V, Bonora M, Simoneschi D, Sartini D, Kudo Y, Saraf A, 
            Florens L, Washburn MP, Stadtfeld M, Pinton P, Pagano M
  TITLE     The TDH-GCN5L1-Fbxo15-KBP axis limits mitochondrial biogenesis in 
            mouse embryonic stem cells.
  JOURNAL   Nat Cell Biol. 2017 Apr;19(4):341-351. doi: 10.1038/ncb3491. Epub 
            2017 Mar 20.
  PUBMED    28319092
REFERENCE   2  (bases 1 to 6716)
  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: "10.1038/ncb3491"; 
            journalName: "Nat Cell Biol"; date: "2017-04"; volume: "19"; issue: 
            "4"; pages: "341-351"
COMMENT     Sequence Label: pcDNA3 FLAG FBXO15
FEATURES             Location/Qualifiers
     source          1..6716
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     404..423
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(642..746)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(821..840)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1012..1391
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1392..1595
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1546..1566
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1640..1659
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1640..1658
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2992..3015
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(3046..3064)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3047..3064)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3084..3101)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3090..3314
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3360..3788
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3447..3466)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3657..3678
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3797..3817)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3802..4131
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3982..4117
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4044..4063
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4198..4992
                     /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(4252..4271)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4862..4881
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5166..5287
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5203..5222)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5257..5276
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5336..5352)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5336..5352)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5349..5371)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5360..5376
                     /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(5384..5390,5391..5408,5409..5414))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5429..5450
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5567..5584)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5738..6326)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5818..5837)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6497..6716,1..572,573..641))
                     /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"
ORIGIN
        1 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
       61 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
      121 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
      181 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
      241 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
      301 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
      361 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
      421 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
      481 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
      541 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
      601 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
      661 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
      721 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
      781 ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg
      841 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
      901 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
      961 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat
     1021 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     1081 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     1141 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     1201 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     1261 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     1321 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     1381 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
     1441 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     1501 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
     1561 ggcgtgtacg gtgggaggtc tatataagca gagctctctg gctaactaga gaacccactg
     1621 cttactggct tatcgaaatt aatacgactc actataggga gacccaagct tggtaccgag
     1681 ctcggatcca tgccttcaga aatcttgctg aagatatttt cctacttgga tgctgtgagc
     1741 cttctgtgta ctggatgtgt gagcaggcgc ttttatcatc tagccaatga caattttatt
     1801 tggatcggaa tctactcaac tgctttttca cctgcaagat caaattggaa atttaattca
     1861 gtagagaaga tagctatgtc tatgagcttt ctgtcagttc aggataaaga agctggttat
     1921 tggaagaaag aatatatcac aaaacaaata gcatctgtaa aagccgcact agctgacatt
     1981 ctcaaacctg tcaaccctta cacaggcctt ccagttaaga ccaaagaggc cctcagaata
     2041 tttggtttag gttgggcaat tatactgaaa gaaaaaggtg gaaaagaata tatcatggag
     2101 catgttgatc tttccataaa tgacacatca gttactgtta tatggtatgg caaaaaatgg
     2161 ccatgcctag catcattgtc aaccttagat ttatgtggca tgacaccagt ttttaccgac
     2221 tggtataaaa ctcccaccaa acatagactc cgatggcatt ctttaattgc aaagtacaat
     2281 ctgagtcatt tgaccatatc taccatgatt ggctgtgaca gactcattcg gatcttctgc
     2341 ctgcaccctg gcctcctggt gggagtgtgg aagaaggagg aagaactggc ttttgttatg
     2401 gcaaatcttc attttcatca ccttgtggag aggagcacat taggctcggc tactatcccc
     2461 tatgaactgc ctccacatag cccctttttg gatgatagcc ccgagtatgg actgcacggc
     2521 taccaactcc atgttgatct gcacagcggt ggggttttct acctatgtgg tacatttcgc
     2581 aatctcttca ccaagagagg aaatattgaa aatggacatg tgaagctcat tgttatacat
     2641 ttaaaaaata acagagaaca cctacctctt attggaaaag ttggcctctc gtggaaaact
     2701 gatatttttg atggctgtat aaagagttgt tccatgatgg acgtaactct tttggatgaa
     2761 catgggaaac ccttttggtg tttcagttcc ccggtgtgcc tgagatcgcc tgccacaccc
     2821 tctgacagct ctagcttctt gggacagaca tacaacgtgg actacgttga tgcggaagga
     2881 agagtgcacg tggagctggt gtggatcaga gagaccgaag aataccttat tgtcaacctg
     2941 gtcctttatc ttagtatcgc aaaaatcaac cattggtttg ggactgaata tgactacaag
     3001 gacgacgatg acaagtagct cgagcatgca tctagagggc cctattctat agtgtcacct
     3061 aaatgctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     3121 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     3181 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     3241 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     3301 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     3361 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3421 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3481 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     3541 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3601 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3661 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3721 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3781 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3841 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3901 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3961 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4021 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     4081 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     4141 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     4201 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4261 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4321 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     4381 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4441 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4501 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4561 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4621 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4681 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     4741 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     4801 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     4861 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4921 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     4981 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     5041 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     5101 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     5161 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5221 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5281 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     5341 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     5401 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     5461 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5521 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5581 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5641 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5701 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5761 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5821 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5881 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5941 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6001 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6061 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6121 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6181 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6241 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6301 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6361 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6421 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6481 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6541 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6601 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6661 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgca
//
