LOCUS       sequence-249979-        6890 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses myc-tagged NDUFS2/H60A 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 H60A/myc-His
FEATURES             Location/Qualifiers
     source          1..6890
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        85..464
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        465..668
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     619..639
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     713..732
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        713..731
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2225..2254
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2270..2287
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2331..2348)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2337..2561
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2607..3035
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2694..2713)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2904..2925
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3044..3064)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3049..3378
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3229..3364
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3291..3310
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3445..4239
                     /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(3499..3518)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4109..4128
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4413..4534
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4450..4469)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4504..4523
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4583..4599)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4583..4599)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4596..4618)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4607..4623
                     /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(4631..4637,4638..4655,4656..4661))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4676..4697
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4814..4831)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4985..5573)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5065..5084)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5744..6535,6536..6604))
                     /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     6367..6386
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6605..6709)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6784..6803)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg
       61 atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa
      121 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
      181 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      241 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      301 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      361 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      421 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
      481 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
      541 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
      601 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
      661 gcagagctct ctggctaact agagaaccca ctgcttactg gcttatcgaa attaatacga
      721 ctcactatag ggagacccaa gctggctagc taagatggcg gcgctgaggg ctttgtgcgg
      781 cttccggggc gtcgcggccc aggtgctgcg gcctggggct ggagtccgat tgccgattca
      841 gcccagcaga ggtgttcggc agtggcagcc agatgtggaa tgggcacagc agtttggggg
      901 agctgttatg tacccaagca aagaaacagc cgcatggaag cctccacctt ggaatgatgt
      961 ggaccctcca aaggacacaa ttgtgaagaa cattaccctg aactttgggc cccaacaccc
     1021 agcagcgcat ggtgtcctgc gactagtgat ggaattgagt ggggagatgg tgcggaagtg
     1081 tgatcctcac atcgggctcc tgcaccgagg cactgagaag ctcattgaat acaagaccta
     1141 tcttcaggcc cttccatact ttgaccggct agactatgtg tccatgatgt gtaacgaaca
     1201 ggcctattct ctagctgtgg agaagttgct aaacatccgg cctcctcctc gggcacagtg
     1261 gatccgagtg ctgtttggag aaatcacacg tttgttgaac cacatcatgg ctgtgaccac
     1321 acatgccctg gaccttgggg ccatgacccc tttcttctgg ctgtttgaag aaagggagaa
     1381 gatgtttgag ttctacgagc gagtgtctgg agcccgaatg catgctgctt atatccggcc
     1441 aggaggagtg caccaggacc taccccttgg gcttatggat gacatttatc agttttctaa
     1501 gaacttctct cttcggcttg atgagttgga ggagttgctg accaacaata ggatctggcg
     1561 aaatcggaca attgacattg gggttgtaac agcagaagaa gcacttaact atggttttag
     1621 tggagtgatg cttcggggct caggcatcca gtgggacctg cggaagaccc agccctatga
     1681 tgtttacgac caggttgagt ttgatgttcc tgttggttct cgaggggact gctatgatag
     1741 gtacctgtgc cgggtggagg agatgcgcca gtccctgaga attatcgcac agtgtctaaa
     1801 caagatgcct cctggggaga tcaaggttga tgatgccaaa gtgtctccac ctaagcgagc
     1861 agagatgaag acttccatgg agtcactgat tcatcacttt aagttgtata ctgagggcta
     1921 ccaagttcct ccaggagcca catatactgc cattgaggct cccaagggag agtttggggt
     1981 gtacctggtg tctgatggca gcagccgccc ttatcgatgc aagatcaagg ctcctggttt
     2041 tgcccatctg gctggtttgg acaagatgtc taagggacac atgttggcag atgtcgttgc
     2101 catcataggt acccaagata ttgtatttgg agaagtagat cgggcggccg ccactgtgct
     2161 ggatatctgc agaattccac cacactggac tagtggatcc gagctcggta ccaagcttgg
     2221 gcccgaacaa aaactcatct cagaagagga tctgaatagc gccgtcgacc atcatcatca
     2281 tcatcattga gtttaaacgg tctccagctt aagtttaaac cgctgatcag cctcgactgt
     2341 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     2401 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     2461 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     2521 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     2581 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     2641 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     2701 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     2761 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     2821 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     2881 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     2941 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     3001 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     3061 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3121 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3181 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3241 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3301 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3361 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     3421 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     3481 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     3541 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     3601 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     3661 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     3721 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     3781 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     3841 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     3901 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     3961 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4021 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4081 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4141 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4201 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     4261 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     4321 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     4381 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     4441 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     4501 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     4561 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     4621 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     4681 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     4741 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     4801 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     4861 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4921 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4981 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5041 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5101 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5161 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5221 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5281 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5341 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5401 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     5461 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5521 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5581 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5641 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     5701 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     5761 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     5821 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     5881 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5941 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6001 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6061 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6121 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6181 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6241 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6301 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6361 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6421 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6481 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6541 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6601 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     6661 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     6721 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
     6781 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     6841 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag
//
