LOCUS       sequence-249976-        6890 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses myc-tagged NDUFS2/P63A 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 P63A/myc-His
FEATURES             Location/Qualifiers
     source          1..6890
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        130..509
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        510..713
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     664..684
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     758..777
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        758..776
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2270..2299
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2315..2332
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2376..2393)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2382..2606
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2652..3080
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2739..2758)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2949..2970
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3089..3109)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3094..3423
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3274..3409
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3336..3355
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3490..4284
                     /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(3544..3563)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4154..4173
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4458..4579
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4495..4514)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4549..4568
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4628..4644)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4628..4644)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4641..4663)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4652..4668
                     /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(4676..4682,4683..4700,4701..4706))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4721..4742
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4859..4876)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5030..5618)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5110..5129)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5789..6580,6581..6649))
                     /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     6412..6431
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6650..6754)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6829..6848)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
       61 gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat
      121 atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
      181 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      241 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      301 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      361 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      421 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      481 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      541 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      601 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      661 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc
      721 taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga
      781 cccaagctgg ctagctaaga tggcggcgct gagggctttg tgcggcttcc ggggcgtcgc
      841 ggcccaggtg ctgcggcctg gggctggagt ccgattgccg attcagccca gcagaggtgt
      901 tcggcagtgg cagccagatg tggaatgggc acagcagttt gggggagctg ttatgtaccc
      961 aagcaaagaa acagcccact ggaaggcgcc accttggaat gatgtggacc ctccaaagga
     1021 cacaattgtg aagaacatta ccctgaactt tgggccccaa cacccagcag cgcatggtgt
     1081 cctgcgacta gtgatggaat tgagtgggga gatggtgcgg aagtgtgatc ctcacatcgg
     1141 gctcctgcac cgaggcactg agaagctcat tgaatacaag acctatcttc aggcccttcc
     1201 atactttgac cggctagact atgtgtccat gatgtgtaac gaacaggcct attctctagc
     1261 tgtggagaag ttgctaaaca tccggcctcc tcctcgggca cagtggatcc gagtgctgtt
     1321 tggagaaatc acacgtttgt tgaaccacat catggctgtg accacacatg ccctggacct
     1381 tggggccatg acccctttct tctggctgtt tgaagaaagg gagaagatgt ttgagttcta
     1441 cgagcgagtg tctggagccc gaatgcatgc tgcttatatc cggccaggag gagtgcacca
     1501 ggacctaccc cttgggctta tggatgacat ttatcagttt tctaagaact tctctcttcg
     1561 gcttgatgag ttggaggagt tgctgaccaa caataggatc tggcgaaatc ggacaattga
     1621 cattggggtt gtaacagcag aagaagcact taactatggt tttagtggag tgatgcttcg
     1681 gggctcaggc atccagtggg acctgcggaa gacccagccc tatgatgttt acgaccaggt
     1741 tgagtttgat gttcctgttg gttctcgagg ggactgctat gataggtacc tgtgccgggt
     1801 ggaggagatg cgccagtccc tgagaattat cgcacagtgt ctaaacaaga tgcctcctgg
     1861 ggagatcaag gttgatgatg ccaaagtgtc tccacctaag cgagcagaga tgaagacttc
     1921 catggagtca ctgattcatc actttaagtt gtatactgag ggctaccaag ttcctccagg
     1981 agccacatat actgccattg aggctcccaa gggagagttt ggggtgtacc tggtgtctga
     2041 tggcagcagc cgcccttatc gatgcaagat caaggctcct ggttttgccc atctggctgg
     2101 tttggacaag atgtctaagg gacacatgtt ggcagatgtc gttgccatca taggtaccca
     2161 agatattgta tttggagaag tagatcgggc ggccgccact gtgctggata tctgcagaat
     2221 tccaccacac tggactagtg gatccgagct cggtaccaag cttgggcccg aacaaaaact
     2281 catctcagaa gaggatctga atagcgccgt cgaccatcat catcatcatc attgagttta
     2341 aacggtctcc agcttaagtt taaaccgctg atcagcctcg actgtgcctt ctagttgcca
     2401 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     2461 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     2521 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     2581 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     2641 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     2701 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     2761 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     2821 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     2881 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     2941 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     3001 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     3061 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3121 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3181 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3241 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3301 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3361 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3421 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     3481 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     3541 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     3601 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     3661 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     3721 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     3781 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     3841 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     3901 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     3961 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     4021 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4081 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4141 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     4201 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4261 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     4321 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4381 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4441 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4501 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4561 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     4621 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4681 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4741 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4801 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4861 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4921 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4981 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5041 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5101 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5161 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5221 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5281 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5341 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5401 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5461 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5521 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5581 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5641 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5701 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5761 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     5821 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5881 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5941 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6001 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6061 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6121 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6181 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6241 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6301 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6361 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6421 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6481 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6541 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6601 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6661 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6721 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     6781 acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc
     6841 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
//
