LOCUS       sequence-231820-        6126 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression in E.coli and affinity purification of N-terminally 
            His tagged NQO1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6126)
  AUTHORS   Tsvetkov P, Asher G, Reiss V, Shaul Y, Sachs L, Lotem J
  TITLE     Inhibition of NAD(P)H:quinone oxidoreductase 1 activity and 
            induction of p53 degradation by the natural phenolic compound 
            curcumin.
  JOURNAL   Proc Natl Acad Sci U S A. 2005 Apr 12;102(15):5535-40. doi: 
            10.1073/pnas.0501828102. Epub 2005 Apr 4.
  PUBMED    15809436
REFERENCE   2  (bases 1 to 6126)
  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: 
            "10.1073/pnas.0501828102"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2005-04-12- 12"; volume: "102"; issue: "15"; pages: "5535-40"
COMMENT     Sequence Label: pET28-His-TEV-NQO1
FEATURES             Location/Qualifiers
     source          1..6126
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     106..125
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     209..228
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        209..227
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    228..252
                     /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)."
     RBS             267..289
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             308..325
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             335..355
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1199..1216
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1269..1287)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1283..1330
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1367..1822
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1454..1473)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1664..1685
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1914..2729)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1914..2729)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2637..2656
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2721..2740
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2851..3439
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3340..3359
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3593..3610
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3625..3764
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3690..3709)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3850..3872
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3866..4057)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4866..5948)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5910..5929
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5949..6026)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa
       61 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
      121 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
      181 gaggatcgag atctcgatcc cgcgaaatta atacgactca ctatagggga attgtgagcg
      241 gataacaatt cccctctaga aataattttg tttaacttta agaaggagat ataccatggg
      301 cagcagccat catcatcatc atcactccgc gggtgaaaac ctgtacttcc agggtgtcgg
      361 cagaagagca ctgatcgtac tggctcactc agagaggacc tccttcaact atgccatgaa
      421 ggaggctgct gcagcggctt tgaagaagaa aggatgggag gtggtggagt cggacctcta
      481 tgccatgaac ttcaatccca tcatttccag aaaggacatc acaggtaaac tgaaggaccc
      541 tgcgaacttt cagtatcctg ccgagtctgt tctggcttat aaagaaggcc atctgagccc
      601 agatattgtg gctgaacaaa agaagctgga agccgcagac cttgtgatat tccagttccc
      661 cctgcagtgg tttggagtcc ctgccattct gaaaggctgg tttgagcgag tgttcatagg
      721 agagtttgct tacacttacg ctgccatgta tgacaaagga cccttccgga gtaagaaggc
      781 agtgctttcc atcaccactg gtggcagtgg ctccatgtac tctctgcaag ggatccacgg
      841 ggacatgaat gtcattctct ggccaattca gagtggcatt ctgcatttct gtggcttcca
      901 agtcttagaa cctcaactga catatagcat tgggcacact ccagcagacg cccgaattca
      961 aatcctggaa ggatggaaga aacgcctgga gaatatttgg gatgagacac cactgtattt
     1021 tgctccaagc agcctctttg acctaaactt ccaggcagga ttcttaatga aaaaagaggt
     1081 acaggatgag gagaaaaaca agaaatttgg cctttctgtg ggccatcact tgggcaagtc
     1141 catcccaact gacaaccaga tcaaagctag aaaataaaag cttgcggccg cactcgagca
     1201 ccaccaccac caccactgag atccggctgc taacaaagcc cgaaaggaag ctgagttggc
     1261 tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
     1321 gggttttttg ctgaaaggag gaactatatc cggattggcg aatgggacgc gccctgtagc
     1381 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     1441 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     1501 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     1561 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     1621 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     1681 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     1741 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     1801 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     1861 tttgtttatt tttctaaata cattcaaata tgtatccgct catgaattaa ttcttagaaa
     1921 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat
     1981 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg
     2041 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat
     2101 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
     2161 ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta
     2221 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga
     2281 gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac
     2341 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct
     2401 aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga
     2461 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg
     2521 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
     2581 ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg
     2641 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggcctagag
     2701 caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca
     2761 gacagtttta ttgttcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     2821 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     2881 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2941 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     3001 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3061 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3121 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3181 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3241 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3301 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     3361 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     3421 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3481 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     3541 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     3601 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     3661 tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     3721 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
     3781 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     3841 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     3901 ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt
     3961 catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc
     4021 gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat
     4081 ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta
     4141 ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga
     4201 tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg
     4261 taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc
     4321 agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg
     4381 ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg
     4441 gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca
     4501 ggagcacgat catgcgcacc cgtggggccg ccatgccggc gataatggcc tgcttctcgc
     4561 cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga
     4621 ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa
     4681 tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa
     4741 gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc
     4801 tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt
     4861 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4921 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc
     4981 agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag
     5041 cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg
     5101 atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg
     5161 cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc
     5221 agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac
     5281 atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat
     5341 ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc
     5401 gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca
     5461 tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga
     5521 acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg
     5581 atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg
     5641 ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta
     5701 atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc
     5761 agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc
     5821 gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc
     5881 acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact
     5941 ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga
     6001 aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg
     6061 cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg
     6121 tgcatg
//
