LOCUS       sequence-350667-        5810 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial vector for expression of an N-terminal GST fusion of 
            NQO1(Y128F) with a TEV protease site located between the GST tag and
            NQO1(Y128F).
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(Y128F)
FEATURES             Location/Qualifiers
     source          1..5810
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(559..564,565..582,583..589)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    597..613
                     /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     621..637
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     621..637
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             633..1160
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(653..670)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(653..669)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(662..684)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(1262..1267,1268..1283,1284..1290)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1298..1314
                     /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             1337..1990
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1948..1970
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1997..2014
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2015..2035
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(2939..2961)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3106..3124)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3192..3296
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3297..3365,3366..4157)
                     /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(3515..3534)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4328..4916
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4817..4836
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5070..5087
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5160..5237
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5238..5810,1..510)
                     /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(5257..5276)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag
       61 ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa
      121 atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg
      181 ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg
      241 ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgttaaccac catcaaacag
      301 gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag
      361 gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac caccctggcg
      421 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
      481 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtaa gttagctcac
      541 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
      601 gagcggataa caatttcaca caggaaacag ctatgaccat gattacggat tcactggccg
      661 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
      721 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
      781 aacagttgcg cagcctgaat ggcgaatggc gctttgcctg gtttccggca ccagaagcgg
      841 tgccggaaag ctggctggag tgcgatcttc ctgaggccga tactgtcgtc gtcccctcaa
      901 actggcagat gcacggttac gatgcgccca tctacaccaa cgtgacctat cccattacgg
      961 tcaatccgcc gtttgttccc acggagaatc cgacgggttg ttactcgctc acatttaatg
     1021 ttgatgaaag ctggctacag gaaggccaga cgcgaattat ttttgatggc gttggaatta
     1081 gcttatcgac tgcacggtgc accaatgctt ctggcgtcag gcagccatcg gaagctgtgg
     1141 tatggctgtg caggtcgtaa atcactgcat aattcgtgtc gctcaaggcg cactcccgtt
     1201 ctggataatg ttttttgcgc cgacatcata acggttctgg caaatattct gaaatgagct
     1261 gttgacaatt aatcatcggc tcgtataatg tgtggaattg tgagcggata acaatttcac
     1321 acaggaaaca gtattcatgt cccctatact aggttattgg aaaattaagg gccttgtgca
     1381 acccactcga cttcttttgg aatatcttga agaaaaatat gaagagcatt tgtatgagcg
     1441 cgatgaaggt gataaatggc gaaacaaaaa gtttgaattg ggtttggagt ttcccaatct
     1501 tccttattat attgatggtg atgttaaatt aacacagtct atggccatca tacgttatat
     1561 agctgacaag cacaacatgt tgggtggttg tccaaaagag cgtgcagaga tttcaatgct
     1621 tgaaggagcg gttttggata ttagatacgg tgtttcgaga attgcatata gtaaagactt
     1681 tgaaactctc aaagttgatt ttcttagcaa gctacctgaa atgctgaaaa tgttcgaaga
     1741 tcgtttatgt cataaaacat atttaaatgg tgatcatgta acccatcctg acttcatgtt
     1801 gtatgacgct cttgatgttg ttttatacat ggacccaatg tgcctggatg cgttcccaaa
     1861 attagtttgt tttaaaaaac gtattgaagc tatcccacaa attgataagt acttgaaatc
     1921 cagcaagtat atagcatggc ctttgcaggg ctggcaagcc acgtttggtg gtggcgacca
     1981 tcctccaaaa tcggatctgg ttccgcgtgg atccgaaaac ctgtattttc agggctcgaa
     2041 ttcccgggtc gacatggtcg gcagaagagc actgatcgta ctggctcact cagagaggac
     2101 ctccttcaac tatgccatga aggaggctgc tgcagcggct ttgaagaaga aaggatggga
     2161 ggtggtggag tcggacctct atgccatgaa cttcaatccc atcatttcca gaaaggacat
     2221 cacaggtaaa ctgaaggacc ctgcgaactt tcagtatcct gccgagtctg ttctggctta
     2281 taaagaaggc catctgagcc cagatattgt ggctgaacaa aagaagctgg aagccgcaga
     2341 ccttgtgata ttccagttcc ccctgcagtg gtttggagtc cctgccattc tgaaaggctg
     2401 gtttgagcga gtgttcatag gagagtttgc ttacactttc gctgccatgt atgacaaagg
     2461 acccttccgg agtaagaagg cagtgctttc catcaccact ggtggcagtg gctccatgta
     2521 ctctctgcaa gggatccacg gggacatgaa tgtcattctc tggccaattc agagtggcat
     2581 tctgcatttc tgtggcttcc aagtcttaga acctcaactg acatatagca ttgggcacac
     2641 tccagcagac gcccgaattc aaatcctgga aggatggaag aaacgcctgg agaatatttg
     2701 ggatgagaca ccactgtatt ttgctccaag cagcctcttt gacctaaact tccaggcagg
     2761 attcttaatg aaaaaagagg tacaggatga ggagaaaaac aagaaatttg gcctttctgt
     2821 gggccatcac ttgggcaagt ccatcccaac tgacaaccag atcaaagcta gaaaatgagc
     2881 ggccgcatcg tgactgactg acgatctgcc tcgcgcgttt cggtgatgac ggtgaaaacc
     2941 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     3001 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc
     3061 agtcacgtag cgatagcgga gtgtataatt cttgaagacg aaagggcctc gtgatacgcc
     3121 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     3181 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3241 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     3301 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     3361 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     3421 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     3481 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
     3541 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     3601 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     3661 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     3721 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     3781 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     3841 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     3901 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     3961 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     4021 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     4081 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     4141 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     4201 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4261 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4321 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4381 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4441 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     4501 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4561 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4621 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4681 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4741 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4801 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4861 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4921 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4981 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     5041 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     5101 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc ataaattccg
     5161 acaccatcga atggcgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc
     5221 aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg
     5281 tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc
     5341 gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac
     5401 aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg
     5461 cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg
     5521 tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc
     5581 tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg
     5641 ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac
     5701 ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg gagcatctgg
     5761 tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct
//
