LOCUS       sequence-350666-        5810 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial vector for expression of an N-terminal GST fusion of NQO1 
            with a TEV protease site located between the GST tag and NQO1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5810)
  AUTHORS   Fang Q, Andrews J, Sharma N, Wilk A, Clark J, Slyskova J, Koczor CA,
            Lans H, Prakash A, Sobol RW
  TITLE     Stability and sub-cellular localization of DNA polymerase beta is 
            regulated by interactions with NQO1 and XRCC1 in response to 
            oxidative stress.
  JOURNAL   Nucleic Acids Res. 2019 Jul 9;47(12):6269-6286. doi: 
            10.1093/nar/gkz293.
  PUBMED    31287140
REFERENCE   2  (bases 1 to 5810)
  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"; journalName: 
            "Nucleic Acids Res"; date: "2019-07-9- 9"; volume: "47"; issue: 
            "12"; pages: "6269-6286"
COMMENT     Sequence Label: pGEX4T3-GST-NQO1
FEATURES             Location/Qualifiers
     source          1..5810
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(529..534,535..552,553..559)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    567..583
                     /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)."
     primer_bind     591..607
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     591..607
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             603..1130
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(623..640)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(623..639)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(632..654)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(1232..1237,1238..1253,1254..1260)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1268..1284
                     /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)."
     CDS             1307..1960
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1918..1940
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1967..1984
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1985..2005
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(2909..2931)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3076..3094)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3162..3266
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3267..3335,3336..4127)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3485..3504)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4298..4886
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4787..4806
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5040..5057
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5130..5207
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5208..5810,1..480)
                     /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     complement(5227..5246)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 cataaatatc tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg cgactggagt
       61 gccatgtccg gttttcaaca aaccatgcaa atgctgaatg agggcatcgt tcccactgcg
      121 atgctggttg ccaacgatca gatggcgctg ggcgcaatgc gcgccattac cgagtccggg
      181 ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg ataccgaaga cagctcatgt
      241 tatatcccgc cgttaaccac catcaaacag gattttcgcc tgctggggca aaccagcgtg
      301 gaccgcttgc tgcaactctc tcagggccag gcggtgaagg gcaatcagct gttgcccgtc
      361 tcactggtga aaagaaaaac caccctggcg cccaatacgc aaaccgcctc tccccgcgcg
      421 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
      481 gcgcaacgca attaatgtaa gttagctcac tcattaggca ccccaggctt tacactttat
      541 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
      601 ctatgaccat gattacggat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
      661 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
      721 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc
      781 gctttgcctg gtttccggca ccagaagcgg tgccggaaag ctggctggag tgcgatcttc
      841 ctgaggccga tactgtcgtc gtcccctcaa actggcagat gcacggttac gatgcgccca
      901 tctacaccaa cgtgacctat cccattacgg tcaatccgcc gtttgttccc acggagaatc
      961 cgacgggttg ttactcgctc acatttaatg ttgatgaaag ctggctacag gaaggccaga
     1021 cgcgaattat ttttgatggc gttggaatta gcttatcgac tgcacggtgc accaatgctt
     1081 ctggcgtcag gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa atcactgcat
     1141 aattcgtgtc gctcaaggcg cactcccgtt ctggataatg ttttttgcgc cgacatcata
     1201 acggttctgg caaatattct gaaatgagct gttgacaatt aatcatcggc tcgtataatg
     1261 tgtggaattg tgagcggata acaatttcac acaggaaaca gtattcatgt cccctatact
     1321 aggttattgg aaaattaagg gccttgtgca acccactcga cttcttttgg aatatcttga
     1381 agaaaaatat gaagagcatt tgtatgagcg cgatgaaggt gataaatggc gaaacaaaaa
     1441 gtttgaattg ggtttggagt ttcccaatct tccttattat attgatggtg atgttaaatt
     1501 aacacagtct atggccatca tacgttatat agctgacaag cacaacatgt tgggtggttg
     1561 tccaaaagag cgtgcagaga tttcaatgct tgaaggagcg gttttggata ttagatacgg
     1621 tgtttcgaga attgcatata gtaaagactt tgaaactctc aaagttgatt ttcttagcaa
     1681 gctacctgaa atgctgaaaa tgttcgaaga tcgtttatgt cataaaacat atttaaatgg
     1741 tgatcatgta acccatcctg acttcatgtt gtatgacgct cttgatgttg ttttatacat
     1801 ggacccaatg tgcctggatg cgttcccaaa attagtttgt tttaaaaaac gtattgaagc
     1861 tatcccacaa attgataagt acttgaaatc cagcaagtat atagcatggc ctttgcaggg
     1921 ctggcaagcc acgtttggtg gtggcgacca tcctccaaaa tcggatctgg ttccgcgtgg
     1981 atccgaaaac ctgtattttc agggctcgaa ttcccgggtc gacatggtcg gcagaagagc
     2041 actgatcgta ctggctcact cagagaggac ctccttcaac tatgccatga aggaggctgc
     2101 tgcagcggct ttgaagaaga aaggatggga ggtggtggag tcggacctct atgccatgaa
     2161 cttcaatccc atcatttcca gaaaggacat cacaggtaaa ctgaaggacc ctgcgaactt
     2221 tcagtatcct gccgagtctg ttctggctta taaagaaggc catctgagcc cagatattgt
     2281 ggctgaacaa aagaagctgg aagccgcaga ccttgtgata ttccagttcc ccctgcagtg
     2341 gtttggagtc cctgccattc tgaaaggctg gtttgagcga gtgttcatag gagagtttgc
     2401 ttacacttac gctgccatgt atgacaaagg acccttccgg agtaagaagg cagtgctttc
     2461 catcaccact ggtggcagtg gctccatgta ctctctgcaa gggatccacg gggacatgaa
     2521 tgtcattctc tggccaattc agagtggcat tctgcatttc tgtggcttcc aagtcttaga
     2581 acctcaactg acatatagca ttgggcacac tccagcagac gcccgaattc aaatcctgga
     2641 aggatggaag aaacgcctgg agaatatttg ggatgagaca ccactgtatt ttgctccaag
     2701 cagcctcttt gacctaaact tccaggcagg attcttaatg aaaaaagagg tacaggatga
     2761 ggagaaaaac aagaaatttg gcctttctgt gggccatcac ttgggcaagt ccatcccaac
     2821 tgacaaccag atcaaagcta gaaaatgagc ggccgcatcg tgactgactg acgatctgcc
     2881 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     2941 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     3001 ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtataatt
     3061 cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     3121 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3181 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3241 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     3301 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     3361 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     3421 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     3481 ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc
     3541 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     3601 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     3661 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     3721 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     3781 caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat
     3841 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     3901 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     3961 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     4021 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     4081 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     4141 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4201 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4261 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4321 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4381 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4441 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4501 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4561 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4621 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4681 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4741 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     4801 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     4861 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     4921 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     4981 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     5041 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     5101 tgtgcggtat ttcacaccgc ataaattccg acaccatcga atggcgcaaa acctttcgcg
     5161 gtatggcatg atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa
     5221 cgttatacga tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga
     5281 accaggccag ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc
     5341 tgaattacat tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg
     5401 gcgttgccac ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat
     5461 ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg
     5521 aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta
     5581 actatccgct ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg
     5641 cgttatttct tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag
     5701 acggtacgcg actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt
     5761 tagcgggccc attaagttct gtctcggcgc gtctgcgtct ggctggctgg
//
