LOCUS       sequence-244118-        6880 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses myc-tagged NDUFS2/T58A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6880)
  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 6880)
  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 T58A/myc-His
FEATURES             Location/Qualifiers
     source          1..6880
                     /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             2365..2394
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2410..2427
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2471..2488)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2477..2701
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2747..3175
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2834..2853)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3044..3065
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3184..3204)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3189..3518
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3369..3504
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3431..3450
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3585..4379
                     /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(3639..3658)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4249..4268
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4553..4674
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4590..4609)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4644..4663
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4723..4739)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4723..4739)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4736..4758)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4747..4763
                     /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(4771..4777,4778..4795,4796..4801))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4816..4837
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4954..4971)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5125..5713)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5205..5224)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5884..6675,6676..6744))
                     /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     6507..6526
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6745..6849)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 gctggtaaga
      901 tggcggcgct gagggctttg tgcggcttcc ggggcgtcgc ggcccaggtg ctgcggcctg
      961 ggctggagtc cgattgccga ttcagcccag cagaggtgtt cggcagtggc agccagatgt
     1021 ggaatgggca cagcagtttg ggggagctgt tatgtaccca agcaaagaag ccgcccactg
     1081 gaagcctcca ccttggaatg atgtggaccc tccaaaggac acaattgtga agaacattac
     1141 cctgaacttt gggccccaac acccagcagc gcatggtgtc ctgcgactag tgatggaatt
     1201 gagtggggag atggtgcgga agtgtgatcc tcacatcggg ctcctgcacc gaggcactga
     1261 gaagctcatt gaatacaaga cctatcttca ggcccttcca tactttgacc ggctagacta
     1321 tgtgtccatg atgtgtaacg aacaggccta ttctctagct gtggagaagt tgctaaacat
     1381 ccggcctcct cctcgggcac agtggatccg agtgctgttt ggagaaatca cacgtttgtt
     1441 gaaccacatc atggctgtga ccacacatgc cctggacctt ggggccatga cccctttctt
     1501 ctggctgttt gaagaaaggg agaagatgtt tgagttctac gagcgagtgt ctggagcccg
     1561 aatgcatgct gcttatatcc ggccaggagg agtgcaccag gacctacccc ttgggcttat
     1621 ggatgacatt tatcagtttt ctaagaactt ctctcttcgg cttgatgagt tggaggagtt
     1681 gctgaccaac aataggatct ggcgaaatcg gacaattgac attggggttg taacagcaga
     1741 agaagcactt aactatggtt ttagtggagt gatgcttcgg ggctcaggca tccagtggga
     1801 cctgcggaag acccagccct tgatgtttac gaccaggttg agtttgatgt tcctgttggt
     1861 tctcgagggg actgctatga taggtacctg tgccgggtgg aggagatgcg ccagtccctg
     1921 agaattatcg cacagtgtct aaacaagatg cctcctgggg agatcaaggt tgatgatgcc
     1981 aaagtgtctc cacctaagcg agcagagatg aagacttcca tggagtcact gattcatcac
     2041 tttaagttgt atactgaggc taccaagttc ctccaggagc cacatatact gccattgagg
     2101 ctcccaaggg agagtttggg gtgtacctgg tgtctgatgg cagcagccgc ccttatcgat
     2161 gcaagatcaa ggctcctggt tttgcccatc tggctggttt ggacaagatg tctaagggac
     2221 acatgttggc agatgtcgtt gccatcatag gtacccaaga tattgtattt ggagaagtag
     2281 atcggggccg ccactgtgct ggatatctgc agaattccac cacactggac tagtggatcc
     2341 gagctcggta ccaagcttgg gcccgaacaa aaactcatct cagaagagga tctgaatagc
     2401 gccgtcgacc atcatcatca tcatcattga gtttaaacgg tctccagctt aagtttaaac
     2461 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     2521 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     2581 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     2641 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     2701 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     2761 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2821 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2881 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     2941 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     3001 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3061 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     3121 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     3181 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     3241 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3301 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3361 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3421 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     3481 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     3541 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     3601 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     3661 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     3721 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     3781 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     3841 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     3901 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     3961 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4021 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4081 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     4141 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4201 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4261 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4321 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     4381 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     4441 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4501 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     4561 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     4621 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     4681 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     4741 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     4801 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     4861 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     4921 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4981 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5041 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5101 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5161 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5221 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5281 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5341 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5401 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5461 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5521 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     5581 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5641 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5701 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5761 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5821 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5881 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5941 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6001 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6061 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6121 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6181 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6241 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6301 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6361 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     6421 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     6481 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     6541 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     6601 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6661 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6721 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6781 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6841 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
