LOCUS       sequence-250182-        6793 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses myc-tagged _MTS NDUFS2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6793)
  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 6793)
  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 _2-33/myc-His
FEATURES             Location/Qualifiers
     source          1..6793
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        106..485
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        486..689
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     640..660
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     734..753
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        734..752
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2149..2178
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2194..2211
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2255..2272)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2261..2485
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2531..2959
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2618..2637)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2828..2849
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2968..2988)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2973..3302
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3153..3288
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3215..3234
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3369..4163
                     /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(3423..3442)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4033..4052
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4337..4458
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4374..4393)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4428..4447
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4507..4523)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4507..4523)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4520..4542)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4531..4547
                     /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(4555..4561,4562..4579,4580..4585))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4600..4621
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4738..4755)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4909..5497)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4989..5008)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5668..6459,6460..6528))
                     /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     6291..6310
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6529..6633)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6708..6727)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta
       61 ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga
      121 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
      181 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      241 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      301 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      361 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      421 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      481 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      541 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      601 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      661 tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact
      721 ggcttatcga aattaatacg actcactata gggagaccca agctgctagc taagatggtt
      781 cggcagtggc agccagatgt ggaatgggca cagcagtttg ggggagctgt tatgtaccca
      841 agcaaagaaa cagcccactg gaagcctcca ccttggaatg atgtggaccc tccaaaggac
      901 acaattgtga agaacattac cctgaacttt gggccccaac acccagcagc gcatggtgtc
      961 ctgcgactag tgatggaatt gagtggggag atggtgcgga agtgtgatcc tcacatcggg
     1021 ctcctgcacc gaggcactga gaagctcatt gaatacaaga cctatcttca ggcccttcca
     1081 tactttgacc ggctagacta tgtgtccatg atgtgtaacg aacaggccta ttctctagct
     1141 gtggagaagt tgctaaacat ccggcctcct cctcgggcac agtggatccg agtgctgttt
     1201 ggagaaatca cacgtttgtt gaaccacatc atggctgtga ccacacatgc cctggacctt
     1261 ggggccatga cccctttctt ctggctgttt gaagaaaggg agaagatgtt tgagttctac
     1321 gagcgagtgt ctggagcccg aatgcatgct gcttatatcc ggccaggagg agtgcaccag
     1381 gacctacccc ttgggcttat ggatgacatt tatcagtttt ctaagaactt ctctcttcgg
     1441 cttgatgagt tggaggagtt gctgaccaac aataggatct ggcgaaatcg gacaattgac
     1501 attggggttg taacagcaga agaagcactt aactatggtt ttagtggagt gatgcttcgg
     1561 ggctcaggca tccagtggga cctgcggaag acccagccct atgatgttta cgaccaggtt
     1621 gagtttgatg ttcctgttgg ttctcgaggg gactgctatg ataggtacct gtgccgggtg
     1681 gaggagatgc gccagtccct gagaattatc gcacagtgtc taaacaagat gcctcctggg
     1741 gagatcaagg ttgatgatgc caaagtgtct ccacctaagc gagcagagat gaagacttcc
     1801 atggagtcac tgattcatca ctttaagttg tatactgagg gctaccaagt tcctccagga
     1861 gccacatata ctgccattga ggctcccaag ggagagtttg gggtgtacct ggtgtctgat
     1921 ggcagcagcc gcccttatcg atgcaagatc aaggctcctg gttttgccca tctggctggt
     1981 ttggacaaga tgtctaaggg acacatgttg gcagatgtcg ttgccatcat aggtacccaa
     2041 gatattgtat ttggagaagt agatcgggcg gccgccactg tgctggatat ctgcagaatt
     2101 ccaccacact ggactagtgg atccgagctc ggtaccaagc ttgggcccga acaaaaactc
     2161 atctcagaag aggatctgaa tagcgccgtc gaccatcatc atcatcatca ttgagtttaa
     2221 acggtctcca gcttaagttt aaaccgctga tcagcctcga ctgtgccttc tagttgccag
     2281 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     2341 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     2401 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     2461 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     2521 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2581 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     2641 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2701 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     2761 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2821 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     2881 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     2941 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3001 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3061 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3121 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3181 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3241 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3301 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     3361 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     3421 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     3481 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     3541 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     3601 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     3661 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     3721 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     3781 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     3841 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     3901 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     3961 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4021 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4081 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4141 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     4201 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     4261 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     4321 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     4381 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     4441 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     4501 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     4561 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     4621 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     4681 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     4741 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     4801 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     4861 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4921 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4981 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5041 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5101 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5161 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5221 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5281 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5341 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5401 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     5461 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     5521 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     5581 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     5641 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     5701 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     5761 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     5821 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     5881 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     5941 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6001 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6061 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6121 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6181 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6241 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     6301 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     6361 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     6421 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     6481 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     6541 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     6601 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     6661 cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct
     6721 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     6781 tgcgcgagca aaa
//
