LOCUS       sequence-249987-        6887 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses myc-tagged NDUFS2/E57A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6887)
  AUTHORS   Jain S, Hu C, Kluza J, Ke W, Tian G, Giurgiu M, Bleilevens A, Campos
            AR, Charbono A, Stickeler E, Maurer J, Holinski-Feder E, Vaisburg A,
            Bureik M, Luo G, Marchetti P, Cheng Y, Wolf DA
  TITLE     Metabolic targeting of cancer by a ubiquinone uncompetitive 
            inhibitor of mitochondrial complex I.
  JOURNAL   Cell Chem Biol. 2021 Nov 23. pii: S2451-9456(21)00479-7. doi: 
            10.1016/j.chembiol.2021.11.002.
  PUBMED    34852219
REFERENCE   2  (bases 1 to 6887)
  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: "Cell Chem 
            Biol. 2021 Nov 23. pii: S2451-9456(21)00479-7. doi: 
            10.1016/j.chembiol.2021.11.002."
COMMENT     Sequence Label: pcDNA3.1/NDUFS2 E57A/myc-His
FEATURES             Location/Qualifiers
     source          1..6887
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        74..453
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        454..657
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     608..628
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     702..721
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        702..720
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2214..2243
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2317..2334)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2323..2547
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2593..3021
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2680..2699)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2890..2911
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3030..3050)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3035..3364
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3215..3350
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3277..3296
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3431..4225
                     /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(3485..3504)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4095..4114
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4399..4520
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4436..4455)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4490..4509
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4569..4585)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4569..4585)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4582..4604)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4593..4609
                     /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(4617..4623,4624..4641,4642..4647))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4662..4683
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4800..4817)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4971..5559)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5051..5070)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5730..6521,6522..6590))
                     /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     6353..6372
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6591..6695)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6770..6789)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
       61 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
      121 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      181 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      241 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      301 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      361 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      421 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      481 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      541 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      601 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc
      661 tggctaacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg
      721 gagacccaag ctggctagct aagatggcgg cgctgagggc tttgtgcggc ttccggggcg
      781 tcgcggccca ggtgctgcgg cctggggctg gagtccgatt gccgattcag cccagcagag
      841 gtgttcggca gtggcagcca gatgtggaat gggcacagca gtttggggga gctgttatgt
      901 acccaagcaa agcgacagcc cactggaagc ctccaccttg gaatgatgtg gaccctccaa
      961 aggacacaat tgtgaagaac attaccctga actttgggcc ccaacaccca gcagcgcatg
     1021 gtgtcctgcg actagtgatg gaattgagtg gggagatggt gcggaagtgt gatcctcaca
     1081 tcgggctcct gcaccgaggc actgagaagc tcattgaata caagacctat cttcaggccc
     1141 ttccatactt tgaccggcta gactatgtgt ccatgatgtg taacgaacag gcctattctc
     1201 tagctgtgga gaagttgcta aacatccggc ctcctcctcg ggcacagtgg atccgagtgc
     1261 tgtttggaga aatcacacgt ttgttgaacc acatcatggc tgtgaccaca catgccctgg
     1321 accttggggc catgacccct ttcttctggc tgtttgaaga aagggagaag atgtttgagt
     1381 tctacgagcg agtgtctgga gcccgaatgc atgctgctta tatccggcca ggaggagtgc
     1441 accaggacct accccttggg cttatggatg acatttatca gttttctaag aacttctctc
     1501 ttcggcttga tgagttggag gagttgctga ccaacaatag gatctggcga aatcggacaa
     1561 ttgacattgg ggttgtaaca gcagaagaag cacttaacta tggttttagt ggagtgatgc
     1621 ttcggggctc aggcatccag tgggacctgc ggaagaccca gccctatgat gtttacgacc
     1681 aggttgagtt tgatgttcct gttggttctc gaggggactg ctatgatagg tacctgtgcc
     1741 gggtggagga gatgcgccag tccctgagaa ttatcgcaca gtgtctaaac aagatgcctc
     1801 ctggggagat caaggttgat gatgccaaag tgtctccacc taagcgagca gagatgaaga
     1861 cttccatgga gtcactgatt catcacttta agttgtatac tgagggctac caagttcctc
     1921 caggagccac atatactgcc attgaggctc ccaagggaga gtttggggtg tacctggtgt
     1981 ctgatggcag cagccgccct tatcgatgca agatcaaggc tcctggtttt gcccatctgg
     2041 ctggtttgga caagatgtct aagggacaca tgttggcaga tgtcgttgcc atcataggta
     2101 cccaagatat tgtatttgga gaagtagatc gggcggccgc cactgtgctg gatatctgca
     2161 gaattccacc acactggact agtggatccg agctcggtac caagcttggg cccgaacaaa
     2221 aactcatctc agaagaggat ctgaatagcg ccgtcgacca tcatcatcat cattgagttt
     2281 aaacggtctc cagcttaagt ttaaaccgct gatcagcctc gactgtgcct tctagttgcc
     2341 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     2401 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     2461 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     2521 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     2581 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     2641 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     2701 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     2761 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     2821 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     2881 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     2941 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     3001 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     3061 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     3121 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3181 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3241 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     3301 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     3361 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg
     3421 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     3481 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     3541 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     3601 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     3661 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     3721 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     3781 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     3841 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     3901 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     3961 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     4021 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     4081 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     4141 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     4201 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     4261 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     4321 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     4381 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     4441 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     4501 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     4561 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     4621 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     4681 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     4741 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     4801 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4861 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4921 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4981 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5041 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5101 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5161 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5221 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5281 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5341 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5401 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5461 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5521 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5581 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5641 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     5701 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     5761 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     5821 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     5881 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5941 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6001 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     6061 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6121 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6181 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6241 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6301 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6361 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6421 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6481 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6541 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6601 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6661 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     6721 gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct
     6781 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     6841 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgac
//
