LOCUS       sequence-128286-        6295 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human NQO1 Flag tagged at C-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6295)
  AUTHORS   Tsvetkov P, Adamovich Y, Elliott E, Shaul Y
  TITLE     E3 ligase STUB1/CHIP regulates NAD(P)H:quinone oxidoreductase 1 
            (NQO1) accumulation in aged brain, a process impaired in certain 
            Alzheimer disease patients.
  JOURNAL   J Biol Chem. 2011 Mar 18;286(11):8839-45. doi: 
            10.1074/jbc.M110.193276. Epub 2011 Jan 10.
  PUBMED    21220432
REFERENCE   2  (bases 1 to 6295)
  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"; doi: 
            "10.1074/jbc.M110.193276"; journalName: "J Biol Chem"; date: 
            "2011-03-18- 18"; volume: "286"; issue: "11"; pages: "8839-45"
COMMENT     Sequence Label: pCDNA3 NQO1-Flag
FEATURES             Location/Qualifiers
     source          1..6295
                     /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"
     regulatory      901..910
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1729..1752
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(1847..1865)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1848..1865)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1885..1902)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1891..2115
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2161..2589
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2248..2267)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2458..2479
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2598..2618)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2603..2933
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2784..2919
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2846..2865
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3000..3794
                     /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(3054..3073)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3664..3683
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3968..4089
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4005..4024)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4059..4078
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4138..4154)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4138..4154)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4151..4173)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4162..4178
                     /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(4186..4192,4193..4210,4211..4216))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4231..4252
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4369..4386)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4540..5128)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4620..4639)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5299..6090,6091..6159))
                     /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     5922..5941
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6160..6264)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gcttggtacc
      901 gccaccatgg tcggcagaag agcactgatc gtactggctc actcagagag gacctccttc
      961 aactatgcca tgaaggaggc tgctgcagcg gctttgaaga agaaaggatg ggaggtggtg
     1021 gagtcggacc tctatgccat gaacttcaat cccatcattt ccagaaagga catcacaggt
     1081 aaactgaagg accctgcgaa ctttcagtat cctgccgagt ctgttctggc ttataaagaa
     1141 ggccatctga gcccagatat tgtggctgaa caaaagaagc tggaagccgc agaccttgtg
     1201 atattccagt tccccctgca gtggtttgga gtccctgcca ttctgaaagg ctggtttgag
     1261 cgagtgttca taggagagtt tgcttacact tacgctgcca tgtatgacaa aggacccttc
     1321 cggagtaaga aggcagtgct ttccatcacc actggtggca gtggctccat gtactctctg
     1381 caagggatcc acggggacat gaatgtcatt ctctggccaa ttcagagtgg cattctgcat
     1441 ttctgtggct tccaagtctt agaacctcaa ctgacatata gcattgggca cactccagca
     1501 gacgcccgaa ttcaaatcct ggaaggatgg aagaaacgcc tggagaatat ttgggatgag
     1561 acaccactgt attttgctcc aagcagcctc tttgacctaa acttccaggc aggattctta
     1621 atgaaaaaag aggtacagga tgaggagaaa aacaagaaat ttggcctttc tgtgggccat
     1681 cacttgggca agtccatccc aactgacaac cagatcaaag ctagaaaaga ctacaaggac
     1741 gacgatgaca agtgaagatc cactagtaac ggccgccagt gtgctggaat tctgcagata
     1801 tccatcacac tggcggccgc tcgagcatgc atctagaggg ccctattcta tagtgtcacc
     1861 taaatgctag agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     1921 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     1981 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     2041 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     2101 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     2161 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     2221 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     2281 cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta
     2341 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     2401 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     2461 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     2521 aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     2581 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     2641 aggcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     2701 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     2761 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     2821 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     2881 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     2941 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     3001 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     3061 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     3121 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     3181 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     3241 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     3301 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     3361 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     3421 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     3481 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     3541 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     3601 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     3661 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     3721 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     3781 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     3841 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     3901 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     3961 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4021 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4081 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     4141 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4201 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4261 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4321 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4381 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4441 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4501 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4561 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4621 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4681 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     4741 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4801 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4861 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4921 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4981 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5041 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5101 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5161 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5221 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5281 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5341 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5401 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5461 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5521 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5581 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5641 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5701 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     5761 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5821 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5881 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5941 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6001 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6061 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6121 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6181 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6241 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
