LOCUS       sequence-189875-        6472 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses WT human NQO1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6472)
  AUTHORS   Asher G, Tsvetkov P, Kahana C, Shaul Y
  TITLE     A mechanism of ubiquitin-independent proteasomal degradation of the 
            tumor suppressors p53 and p73.
  JOURNAL   Genes Dev. 2005 Feb 1;19(3):316-21.
  PUBMED    15687255
REFERENCE   2  (bases 1 to 6472)
  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: "Genes 
            Dev."; date: "2005-02-1"; volume: "19(3)"; pages: "316-21"
COMMENT     Sequence Label: pEFIRES NQO1 WT
FEATURES             Location/Qualifiers
     source          1..6472
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        46..1208
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          277..1199
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1156..1176
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     intron          1247..1379
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1362..1381
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1424..1442
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2301..2867
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2317..2869
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2461..2478)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2688..2707
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2869..3471
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MGTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     3368..3388
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3705..3839
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3742..3761)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3796..3815
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3914..4369
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4001..4020)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4211..4232
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4436..4455)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4555..4577
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4615..4633)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4701..4805
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4806..4874,4875..5666)
                     /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     complement(5024..5043)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5837..6425
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6326..6345
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttttgctcac atggctcgac agatccccca attcaagctt cgtgaggctc cggtgcccgt
       61 cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat
      121 tgaaccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg
      181 ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac
      241 gttctttttc gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc
      301 gggcctggcc tctttacggg ttatggccct tgcgtgcctt gaattacttc cacctggctc
      361 cagtacgtga ttcttgatcc cgagctggag ccaggggcgg gccttgcgct ttaggagccc
      421 cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc gtgcgaatct
      481 ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt taaaattttt
      541 gatgacctgc tgcgacgctt tttttctggc aagatagtct tgtaaatgcg ggccaggatc
      601 tgcacactgg tatttcggtt tttggggccg cgggcggcga cggggcccgt gcgtcccagc
      661 gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga cgggggtagt
      721 ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct
      781 gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg ccgcttcccg
      841 gccctgctcc agggggctca aaatggagga cgcggcgctc gggagagcgg gcgggtgagt
      901 cacccacaca aaggaaaggg gcctttccgt cctcagccgt cgcttcatgt gactccacgg
      961 agtaccgggc gccgtccagg cacctcgatt agttctggag cttttggagt acgtcgtctt
     1021 taggttgggg ggaggggttt tatgcgatgg agtttcccca cactgagtgg gtggagactg
     1081 aagttaggcc agcttggcac ttgatgtaat tctccttgga atttgccctt tttgagtttg
     1141 gatcttggtt cattctcaag cctcagacag tggttcaaag tttttttctt ccatttcagg
     1201 tgtcgtgaac acgggctgca gaagttggtc gtgaggcact gggcaggtaa gtatcaaggt
     1261 tacaagacag gtttaaggag accaatagaa actgggcttg tcgagacaga gaagactctt
     1321 gcgtttctga taggcaccta ttggtcttac tgacatccac tttgcctttc tctccacagg
     1381 tgtccactcc cagttcaatt acagctctta aggctagagt acttaatacg actcactata
     1441 ggctagcctc gaggctatgg tcggcagaag agcactgatc gtactggctc actcagagag
     1501 gacctccttc aactatgcca tgaaggaggc tgctgcagcg gctttgaaga agaaaggatg
     1561 ggaggtggtg gagtcggacc tctatgccat gaacttcaat cccatcattt ccagaaagga
     1621 catcacaggt aaactgaagg accctgcgaa ctttcagtat cctgccgagt ctgttctggc
     1681 ttataaagaa ggccatctga gcccagatat tgtggctgaa caaaagaagc tggaagccgc
     1741 agaccttgtg atattccagt tccccctgca gtggtttgga gtccctgcca ttctgaaagg
     1801 ctggtttgag cgagtgttca taggagagtt tgcttacact tacgctgcca tgtatgacaa
     1861 aggacccttc cggagtaaga aggcagtgct ttccatcacc actggtggca gtggctccat
     1921 gtactctctg caagggatcc acggggacat gaatgtcatt ctctggccaa ttcagagtgg
     1981 cattctgcat ttctgtggct tccaagtctt agaacctcaa ctgacatata gcattgggca
     2041 cactccagca gacgcccgaa ttcaaatcct ggaaggatgg aagaaacgcc tggagaatat
     2101 ttgggatgag acaccactgt attttgctcc aagcagcctc tttgacctaa acttccaggc
     2161 aggattctta atgaaaaaag aggtacagga tgaggagaaa aacaagaaat ttggcctttc
     2221 tgtgggccat cacttgggca agtccatccc aactgacaac cagatcaaag ctagaaaatg
     2281 agcggccgct ctagcccaat tccgcccccc cccctaacgt tactggccga agccgcttgg
     2341 aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca
     2401 atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc
     2461 ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag
     2521 cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg
     2581 gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac
     2641 aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa
     2701 gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc
     2761 tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc
     2821 ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat gggcaccgag
     2881 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt acgcaccctc
     2941 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgacccgga ccgccacatc
     3001 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     3061 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     3121 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     3181 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     3241 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     3301 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc
     3361 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     3421 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg acgcccgccc
     3481 cacgacccgc agcgcccgac cgaaaggagc gcacgacccc atggctccga ccgaagccac
     3541 ccggggcggc cccgccgacc ccgcacccgc ccccgaggcc caccgactct agctagagga
     3601 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     3661 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     3721 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     3781 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcc
     3841 gggctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     3901 gaatggcgaa tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3961 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     4021 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     4081 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     4141 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4201 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     4261 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     4321 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc ctgatgcggt
     4381 attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa
     4441 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     4501 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     4561 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     4621 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     4681 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4741 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4801 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     4861 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     4921 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     4981 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5041 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5101 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5161 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5221 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5281 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5341 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5401 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5461 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5521 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     5581 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     5641 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     5701 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     5761 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     5821 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5881 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5941 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     6001 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6061 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6121 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6181 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6241 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6301 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6361 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6421 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg cc
//
