LOCUS       sequence-349554-        5675 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial vector for expression of an N-terminal GST fusion of the 
            Tev protease.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5675)
  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 5675)
  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-Tev
FEATURES             Location/Qualifiers
     source          1..5675
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(192..197,198..213,214..220)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    228..244
                     /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             267..920
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     878..900
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             927..944
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             948..1655
                     /codon_start=1
                     /product="tobacco etch virus protease (Kapust et al., 
                     2001)"
                     /label=TEV protease
                     /translation="GESLFKGPRDYNPISSTICHLTNESDGHTTSLYGIGFGPFIITNK
                     HLFRRNNGTLLVQSLHGVFKVKNTTTLQQHLIDGRDMIIIRMPKDFPPFPQKLKFREPQ
                     REERICLVTTNFQTKSMSSMVSDTSCTFPSSDGIFWKHWIQTKDGQCGSPLVSTRDGFI
                     VGIHSASNFTNTNNYFTSVPKNFMELLTNQEAQQWVSGWRLNADSVLWGGHKVFMVKPE
                     EPFQPVKEATQLMN"
     primer_bind     complement(1734..1756)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1901..1919)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1987..2091
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2092..2160,2161..2952)
                     /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(2310..2329)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3123..3711
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3612..3631
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3865..3882
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3955..4032
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4033..5115
                     /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(4052..4071)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5164..5169,5170..5187,5188..5194)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5202..5218
                     /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     5226..5242
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5226..5242
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(5238..5675,1..90)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5258..5275)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5258..5274)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5267..5289)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 gttggaatta gcttatcgac tgcacggtgc accaatgctt ctggcgtcag gcagccatcg
       61 gaagctgtgg tatggctgtg caggtcgtaa atcactgcat aattcgtgtc gctcaaggcg
      121 cactcccgtt ctggataatg ttttttgcgc cgacatcata acggttctgg caaatattct
      181 gaaatgagct gttgacaatt aatcatcggc tcgtataatg tgtggaattg tgagcggata
      241 acaatttcac acaggaaaca gtattcatgt cccctatact aggttattgg aaaattaagg
      301 gccttgtgca acccactcga cttcttttgg aatatcttga agaaaaatat gaagagcatt
      361 tgtatgagcg cgatgaaggt gataaatggc gaaacaaaaa gtttgaattg ggtttggagt
      421 ttcccaatct tccttattat attgatggtg atgttaaatt aacacagtct atggccatca
      481 tacgttatat agctgacaag cacaacatgt tgggtggttg tccaaaagag cgtgcagaga
      541 tttcaatgct tgaaggagcg gttttggata ttagatacgg tgtttcgaga attgcatata
      601 gtaaagactt tgaaactctc aaagttgatt ttcttagcaa gctacctgaa atgctgaaaa
      661 tgttcgaaga tcgtttatgt cataaaacat atttaaatgg tgatcatgta acccatcctg
      721 acttcatgtt gtatgacgct cttgatgttg ttttatacat ggacccaatg tgcctggatg
      781 cgttcccaaa attagtttgt tttaaaaaac gtattgaagc tatcccacaa attgataagt
      841 acttgaaatc cagcaagtat atagcatggc ctttgcaggg ctggcaagcc acgtttggtg
      901 gtggcgacca tcctccaaaa tcggatctgg ttccgcgtgg atcccatgga gaaagcttgt
      961 ttaaggggcc gcgtgattac aacccgatat cgagcaccat ttgtcatttg acgaatgaat
     1021 ctgatgggca cacaacatcg ttgtatggta ttggatttgg tcccttcatc attacaaaca
     1081 agcacttgtt tagaagaaat aatggaacac tgttggtcca atcactacat ggtgtattca
     1141 aggtcaagaa caccacgact ttgcaacaac acctcattga tgggagggac atgataatta
     1201 ttcgcatgcc taaggatttc ccaccatttc ctcaaaagct gaaatttaga gagccacaaa
     1261 gggaagagcg catatgtctt gtgacaacca acttccaaac taagagcatg tctagcatgg
     1321 tgtcagacac tagttgcaca ttcccttcat ctgatggcat attctggaag cattggattc
     1381 aaaccaagga tgggcagtgt ggcagtccat tagtatcaac tagagatggg ttcattgttg
     1441 gtatacactc agcatcgaat ttcaccaaca caaacaatta tttcacaagc gtgccgaaaa
     1501 acttcatgga attgttgaca aatcaggagg cgcagcagtg ggttagtggt tggcgattaa
     1561 atgctgactc agtattgtgg gggggccata aagttttcat ggtgaaacct gaagagcctt
     1621 ttcagccagt taaggaagcg actcaactca tgaatcgtcg tcgccgtcgc taagcggccg
     1681 catcgtgact gactgacgat ctgcctcgcg cgtttcggtg atgacggtga aaacctctga
     1741 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     1801 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca
     1861 cgtagcgata gcggagtgta taattcttga agacgaaagg gcctcgtgat acgcctattt
     1921 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     1981 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     2041 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     2101 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     2161 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     2221 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     2281 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
     2341 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     2401 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     2461 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     2521 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     2581 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca
     2641 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     2701 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     2761 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     2821 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     2881 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     2941 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     3001 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     3061 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     3121 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3181 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     3241 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     3301 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     3361 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3421 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     3481 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     3541 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     3601 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     3661 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     3721 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     3781 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     3841 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
     3901 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcataaa ttccgacacc
     3961 atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc
     4021 agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct
     4081 tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa
     4141 aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg
     4201 gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg
     4261 tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg
     4321 tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg
     4381 caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg
     4441 gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc
     4501 aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca
     4561 ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg
     4621 cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg
     4681 gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc
     4741 atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc
     4801 attaccgagt ccgggctgcg cgttggtgcg gatatctcgg tagtgggata cgacgatacc
     4861 gaagacagct catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg
     4921 gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat
     4981 cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc
     5041 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     5101 gaaagcgggc agtgagcgca acgcaattaa tgtaagttag ctcactcatt aggcacccca
     5161 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     5221 tcacacagga aacagctatg accatgatta cggattcact ggccgtcgtt ttacaacgtc
     5281 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     5341 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     5401 tgaatggcga atggcgcttt gcctggtttc cggcaccaga agcggtgccg gaaagctggc
     5461 tggagtgcga tcttcctgag gccgatactg tcgtcgtccc ctcaaactgg cagatgcacg
     5521 gttacgatgc gcccatctac accaacgtga cctatcccat tacggtcaat ccgccgtttg
     5581 ttcccacgga gaatccgacg ggttgttact cgctcacatt taatgttgat gaaagctggc
     5641 tacaggaagg ccagacgcga attatttttg atggc
//
