LOCUS       sequence-249977-        6890 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses myc-tagged NDUFS2/K62A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6890)
  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 6890)
  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 K62A/myc-His
FEATURES             Location/Qualifiers
     source          1..6890
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        118..497
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        498..701
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     652..672
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     746..765
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        746..764
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2258..2287
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2303..2320
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2364..2381)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2370..2594
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2640..3068
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2727..2746)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2937..2958
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3077..3097)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3082..3411
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3262..3397
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3324..3343
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3478..4272
                     /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(3532..3551)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4142..4161
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4446..4567
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4483..4502)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4537..4556
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4616..4632)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4616..4632)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4629..4651)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4640..4656
                     /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(4664..4670,4671..4688,4689..4694))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4709..4730
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4847..4864)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5018..5606)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5098..5117)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5777..6568,6569..6637))
                     /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     6400..6419
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6638..6742)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6817..6836)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
       61 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      121 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      181 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      241 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      301 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      361 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      421 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      481 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      541 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      601 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      661 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta actagagaac
      721 ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc caagctggct
      781 agctaagatg gcggcgctga gggctttgtg cggcttccgg ggcgtcgcgg cccaggtgct
      841 gcggcctggg gctggagtcc gattgccgat tcagcccagc agaggtgttc ggcagtggca
      901 gccagatgtg gaatgggcac agcagtttgg gggagctgtt atgtacccaa gcaaagaaac
      961 agcccactgg gcacctccac cttggaatga tgtggaccct ccaaaggaca caattgtgaa
     1021 gaacattacc ctgaactttg ggccccaaca cccagcagcg catggtgtcc tgcgactagt
     1081 gatggaattg agtggggaga tggtgcggaa gtgtgatcct cacatcgggc tcctgcaccg
     1141 aggcactgag aagctcattg aatacaagac ctatcttcag gcccttccat actttgaccg
     1201 gctagactat gtgtccatga tgtgtaacga acaggcctat tctctagctg tggagaagtt
     1261 gctaaacatc cggcctcctc ctcgggcaca gtggatccga gtgctgtttg gagaaatcac
     1321 acgtttgttg aaccacatca tggctgtgac cacacatgcc ctggaccttg gggccatgac
     1381 ccctttcttc tggctgtttg aagaaaggga gaagatgttt gagttctacg agcgagtgtc
     1441 tggagcccga atgcatgctg cttatatccg gccaggagga gtgcaccagg acctacccct
     1501 tgggcttatg gatgacattt atcagttttc taagaacttc tctcttcggc ttgatgagtt
     1561 ggaggagttg ctgaccaaca ataggatctg gcgaaatcgg acaattgaca ttggggttgt
     1621 aacagcagaa gaagcactta actatggttt tagtggagtg atgcttcggg gctcaggcat
     1681 ccagtgggac ctgcggaaga cccagcccta tgatgtttac gaccaggttg agtttgatgt
     1741 tcctgttggt tctcgagggg actgctatga taggtacctg tgccgggtgg aggagatgcg
     1801 ccagtccctg agaattatcg cacagtgtct aaacaagatg cctcctgggg agatcaaggt
     1861 tgatgatgcc aaagtgtctc cacctaagcg agcagagatg aagacttcca tggagtcact
     1921 gattcatcac tttaagttgt atactgaggg ctaccaagtt cctccaggag ccacatatac
     1981 tgccattgag gctcccaagg gagagtttgg ggtgtacctg gtgtctgatg gcagcagccg
     2041 cccttatcga tgcaagatca aggctcctgg ttttgcccat ctggctggtt tggacaagat
     2101 gtctaaggga cacatgttgg cagatgtcgt tgccatcata ggtacccaag atattgtatt
     2161 tggagaagta gatcgggcgg ccgccactgt gctggatatc tgcagaattc caccacactg
     2221 gactagtgga tccgagctcg gtaccaagct tgggcccgaa caaaaactca tctcagaaga
     2281 ggatctgaat agcgccgtcg accatcatca tcatcatcat tgagtttaaa cggtctccag
     2341 cttaagttta aaccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     2401 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     2461 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     2521 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     2581 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     2641 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2701 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2761 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     2821 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     2881 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     2941 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3001 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3061 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3121 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3181 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3241 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3301 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     3361 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     3421 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     3481 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     3541 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     3601 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     3661 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     3721 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     3781 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     3841 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     3901 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     3961 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     4021 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     4081 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     4141 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4201 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     4261 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     4321 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     4381 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     4441 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     4501 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     4561 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     4621 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     4681 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     4741 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     4801 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     4861 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4921 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4981 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5041 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5101 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5161 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5221 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5281 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5341 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5401 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     5461 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5521 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5581 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5641 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5701 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5761 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5821 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5881 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5941 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6001 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6061 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6121 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6181 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6241 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6301 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6361 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6421 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6481 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6541 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6601 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6661 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6721 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     6781 cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata
     6841 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
//
