LOCUS       sequence-136096-        7475 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human Y128F NQO1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7475)
  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 7475)
  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 Y128F
FEATURES             Location/Qualifiers
     source          1..7475
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        153..1335
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          384..1326
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1283..1303
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     intron          1475..1607
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1590..1609
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1652..1670
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2529..3095
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2689..2706)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2916..2935
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     polyA_signal    3179..3300
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3190..3209)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3244..3263
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3486..3941
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3573..3592)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3783..3804
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4058..4415
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4058..4249
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4081..4101)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4266..4401
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4466..5260
                     /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(4520..4539)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5130..5149
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5324..5372
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5413..5432)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5532..5554
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5592..5610)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5678..5782
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5783..5851,5852..6643)
                     /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(6001..6020)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6814..7402
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7303..7322
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagtaattca tacaaaagga ctcgcccctg ccttggggaa tcccagggac cgtcgttaaa
       61 ctcccactaa cgtagaaccc agagatcgct gcgttcccgc cccctcaccc gcccgctctc
      121 gtcatcactg aggtggagaa gagcatgcgt gaggctccgg tgcccgtcag tgggcagagc
      181 gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga accggtgcct
      241 agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc
      301 ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca
      361 acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct
      421 ttacgggtta tggcccttgc gtgccttgaa ttacttccac gcccctggct gcagtacgtg
      481 attcttgatc ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa
      541 ggagcccctt cgcctcgtgc ttgagttgag gcctggcttg ggcgctgggg ccgccgcgtg
      601 cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa
      661 aatttttgat gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc
      721 caagatctgc acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg
      781 tcccagcgca catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg
      841 gggtagtctc aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc
      901 ccgccctggg cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg
      961 cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg
     1021 ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac
     1081 tccacggagt accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg
     1141 tcgtctttag gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg
     1201 gagactgaag ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt
     1261 gagtttggat cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca
     1321 tttcaggtgt cgtgaactag aagctttatt gcggtagttt atcacagtta aattgctaac
     1381 gcagtcagtg cttctgacac aacagtctcg aacttaagct gcagtgactc tcttaaggta
     1441 gccttgcaga agttggtcgt gaggcactgg gcaggtaagt atcaaggtta caagacaggt
     1501 ttaaggagac caatagaaac tgggcttgtc gagacagaga agactcttgc gtttctgata
     1561 ggcacctatt ggtcttactg acatccactt tgcctttctc tccacaggtg tccactccca
     1621 gttcaattac agctcttaag gctagagtac ttaatacgac tcactatagg ctagcctcga
     1681 ggctatggtc ggcagaagag cactgatcgt actggctcac tcagagagga cctccttcaa
     1741 ctatgccatg aaggaggctg ctgcagcggc tttgaagaag aaaggatggg aggtggtgga
     1801 gtcggacctc tatgccatga acttcaatcc catcatttcc agaaaggaca tcacaggtaa
     1861 actgaaggac cctgcgaact ttcagtatcc tgccgagtct gttctggctt ataaagaagg
     1921 ccatctgagc ccagatattg tggctgaaca aaagaagctg gaagccgcag accttgtgat
     1981 attccagttc cccctgcagt ggtttggagt ccctgccatt ctgaaaggct ggtttgagcg
     2041 agtgttcata ggagagtttg cttacacttt cgctgccatg tatgacaaag gacccttccg
     2101 gagtaagaag gcagtgcttt ccatcaccac tggtggcagt ggctccatgt actctctgca
     2161 agggatccac ggggacatga atgtcattct ctggccaatt cagagtggca ttctgcattt
     2221 ctgtggcttc caagtcttag aacctcaact gacatatagc attgggcaca ctccagcaga
     2281 cgcccgaatt caaatcctgg aaggatggaa gaaacgcctg gagaatattt gggatgagac
     2341 accactgtat tttgctccaa gcagcctctt tgacctaaac ttccaggcag gattcttaat
     2401 gaaaaaagag gtacaggatg aggagaaaaa caagaaattt ggcctttctg tgggccatca
     2461 cttgggcaag tccatcccaa ctgacaacca gatcaaagct agaaaatgag cggccgctct
     2521 agcccaattc cgcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt
     2581 gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc
     2641 ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag
     2701 gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac
     2761 aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc
     2821 tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc
     2881 acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca
     2941 aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt
     3001 gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg
     3061 gggacgtggt tttcctttga aaaacacgat gataagcttg ccacaacccg ggatcctcta
     3121 gagtcgaccc gggcggccgc ttccctttag tgagggttaa tgcttcgagc agacatgata
     3181 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     3241 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     3301 aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt
     3361 taaagcaagt aaaacctcta caaatgtggt aaaatccgat aaggatcgat ccgggctggc
     3421 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     3481 aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3541 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3601 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3661 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     3721 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3781 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     3841 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     3901 atttaacgcg aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc
     3961 cttacgcatc tgtgcggtat ttcacaccgc atacgcggat ctgcgcagca ccatggcctg
     4021 aaataacctc tgaaagagga acttggttag gtaccttctg aggcggaaag aaccagctgt
     4081 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4141 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     4201 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     4261 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4321 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     4381 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct tgattcttct gacacaacag
     4441 tctcgaactt aaggctagag ccaccatgat tgaacaagat ggattgcacg caggttctcc
     4501 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     4561 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     4621 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     4681 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     4741 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     4801 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     4861 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     4921 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     4981 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     5041 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     5101 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     5161 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     5221 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     5281 atgaccgacc aagcgacgcc caacctgcca tcacgatggc cgcaataaaa tatctttatt
     5341 ttcattacat ctgtgtgttg gttttttgtg tgaatcgata gcgataagga tccgcgtatg
     5401 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     5461 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     5521 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     5581 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     5641 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     5701 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     5761 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     5821 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     5881 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     5941 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     6001 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     6061 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     6121 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     6181 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     6241 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     6301 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     6361 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     6421 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     6481 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     6541 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     6601 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     6661 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     6721 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     6781 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6841 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6901 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     6961 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     7021 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     7081 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     7141 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     7201 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     7261 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     7321 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     7381 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     7441 ttgctggcct tttgctcaca tggctcgaca gatct
//
