LOCUS       sequence-249978-        6890 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses myc-tagged NDUFS2/W61A 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 W61A/myc-His
FEATURES             Location/Qualifiers
     source          1..6890
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        120..499
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        500..703
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     654..674
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     748..767
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        748..766
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2260..2289
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2305..2322
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2366..2383)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2372..2596
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2642..3070
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2729..2748)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2939..2960
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3079..3099)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3084..3413
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3264..3399
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3326..3345
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3480..4274
                     /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(3534..3553)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4144..4163
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4448..4569
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4485..4504)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4539..4558
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4618..4634)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4618..4634)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4631..4653)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4642..4658
                     /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(4666..4672,4673..4690,4691..4696))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4711..4732
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4849..4866)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5020..5608)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5100..5119)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5779..6570,6571..6639))
                     /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     6402..6421
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6640..6744)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6819..6838)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
       61 tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg
      121 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
      181 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      241 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      301 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      361 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      421 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      481 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      541 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      601 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      661 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga
      721 acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg
      781 ctagctaaga tggcggcgct gagggctttg tgcggcttcc ggggcgtcgc ggcccaggtg
      841 ctgcggcctg gggctggagt ccgattgccg attcagccca gcagaggtgt tcggcagtgg
      901 cagccagatg tggaatgggc acagcagttt gggggagctg ttatgtaccc aagcaaagaa
      961 acagcccacg cgaagcctcc accttggaat gatgtggacc ctccaaagga cacaattgtg
     1021 aagaacatta ccctgaactt tgggccccaa cacccagcag cgcatggtgt cctgcgacta
     1081 gtgatggaat tgagtgggga gatggtgcgg aagtgtgatc ctcacatcgg gctcctgcac
     1141 cgaggcactg agaagctcat tgaatacaag acctatcttc aggcccttcc atactttgac
     1201 cggctagact atgtgtccat gatgtgtaac gaacaggcct attctctagc tgtggagaag
     1261 ttgctaaaca tccggcctcc tcctcgggca cagtggatcc gagtgctgtt tggagaaatc
     1321 acacgtttgt tgaaccacat catggctgtg accacacatg ccctggacct tggggccatg
     1381 acccctttct tctggctgtt tgaagaaagg gagaagatgt ttgagttcta cgagcgagtg
     1441 tctggagccc gaatgcatgc tgcttatatc cggccaggag gagtgcacca ggacctaccc
     1501 cttgggctta tggatgacat ttatcagttt tctaagaact tctctcttcg gcttgatgag
     1561 ttggaggagt tgctgaccaa caataggatc tggcgaaatc ggacaattga cattggggtt
     1621 gtaacagcag aagaagcact taactatggt tttagtggag tgatgcttcg gggctcaggc
     1681 atccagtggg acctgcggaa gacccagccc tatgatgttt acgaccaggt tgagtttgat
     1741 gttcctgttg gttctcgagg ggactgctat gataggtacc tgtgccgggt ggaggagatg
     1801 cgccagtccc tgagaattat cgcacagtgt ctaaacaaga tgcctcctgg ggagatcaag
     1861 gttgatgatg ccaaagtgtc tccacctaag cgagcagaga tgaagacttc catggagtca
     1921 ctgattcatc actttaagtt gtatactgag ggctaccaag ttcctccagg agccacatat
     1981 actgccattg aggctcccaa gggagagttt ggggtgtacc tggtgtctga tggcagcagc
     2041 cgcccttatc gatgcaagat caaggctcct ggttttgccc atctggctgg tttggacaag
     2101 atgtctaagg gacacatgtt ggcagatgtc gttgccatca taggtaccca agatattgta
     2161 tttggagaag tagatcgggc ggccgccact gtgctggata tctgcagaat tccaccacac
     2221 tggactagtg gatccgagct cggtaccaag cttgggcccg aacaaaaact catctcagaa
     2281 gaggatctga atagcgccgt cgaccatcat catcatcatc attgagttta aacggtctcc
     2341 agcttaagtt taaaccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     2401 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     2461 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     2521 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     2581 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     2641 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     2701 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     2761 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     2821 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     2881 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     2941 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     3001 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3061 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     3121 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3181 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3241 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3301 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     3361 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     3421 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     3481 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     3541 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     3601 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     3661 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     3721 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     3781 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     3841 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     3901 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     3961 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     4021 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     4081 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     4141 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     4201 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     4261 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     4321 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4381 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4441 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4501 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4561 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     4621 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4681 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4741 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4801 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4861 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4921 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4981 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5041 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5101 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5161 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5221 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5281 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5341 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5401 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5461 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5521 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5581 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5641 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5701 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5761 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5821 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5881 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5941 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6001 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6061 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6121 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6181 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6241 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6301 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6361 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6421 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6481 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6541 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6601 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6661 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6721 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     6781 atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca
     6841 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
//
