LOCUS       sequence-372022-        5993 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial vector for expression of an N-terminal GST fusion of 
            PolB(T304I) with a TEV protease site located between the GST tag and
            PolB(T304I).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5993)
  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 5993)
  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-PolB(T304I)
FEATURES             Location/Qualifiers
     source          1..5993
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(121..126,127..144,145..151)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    159..175
                     /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     183..199
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     183..199
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             195..722
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(215..232)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(215..231)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(224..246)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(824..829,830..845,846..852)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    860..876
                     /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             899..1552
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1510..1532
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1559..1576
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1577..1597
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(2684..2706)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2851..2869)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2937..3041
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3042..3110,3111..3902)
                     /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(3260..3279)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4073..4661
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4562..4581
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4815..4832
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4905..4982
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(4983..5993,1..72)
                     /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(5002..5021)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
       61 agcgggcagt gagcgcaacg caattaatgt aagttagctc actcattagg caccccaggc
      121 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
      181 cacaggaaac agctatgacc atgattacgg attcactggc cgtcgtttta caacgtcgtg
      241 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
      301 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
      361 atggcgaatg gcgctttgcc tggtttccgg caccagaagc ggtgccggaa agctggctgg
      421 agtgcgatct tcctgaggcc gatactgtcg tcgtcccctc aaactggcag atgcacggtt
      481 acgatgcgcc catctacacc aacgtgacct atcccattac ggtcaatccg ccgtttgttc
      541 ccacggagaa tccgacgggt tgttactcgc tcacatttaa tgttgatgaa agctggctac
      601 aggaaggcca gacgcgaatt atttttgatg gcgttggaat tagcttatcg actgcacggt
      661 gcaccaatgc ttctggcgtc aggcagccat cggaagctgt ggtatggctg tgcaggtcgt
      721 aaatcactgc ataattcgtg tcgctcaagg cgcactcccg ttctggataa tgttttttgc
      781 gccgacatca taacggttct ggcaaatatt ctgaaatgag ctgttgacaa ttaatcatcg
      841 gctcgtataa tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagtattcat
      901 gtcccctata ctaggttatt ggaaaattaa gggccttgtg caacccactc gacttctttt
      961 ggaatatctt gaagaaaaat atgaagagca tttgtatgag cgcgatgaag gtgataaatg
     1021 gcgaaacaaa aagtttgaat tgggtttgga gtttcccaat cttccttatt atattgatgg
     1081 tgatgttaaa ttaacacagt ctatggccat catacgttat atagctgaca agcacaacat
     1141 gttgggtggt tgtccaaaag agcgtgcaga gatttcaatg cttgaaggag cggttttgga
     1201 tattagatac ggtgtttcga gaattgcata tagtaaagac tttgaaactc tcaaagttga
     1261 ttttcttagc aagctacctg aaatgctgaa aatgttcgaa gatcgtttat gtcataaaac
     1321 atatttaaat ggtgatcatg taacccatcc tgacttcatg ttgtatgacg ctcttgatgt
     1381 tgttttatac atggacccaa tgtgcctgga tgcgttccca aaattagttt gttttaaaaa
     1441 acgtattgaa gctatcccac aaattgataa gtacttgaaa tccagcaagt atatagcatg
     1501 gcctttgcag ggctggcaag ccacgtttgg tggtggcgac catcctccaa aatcggatct
     1561 ggttccgcgt ggatccgaaa acctgtattt tcagggctcg aattcccggg tcgacatgag
     1621 caaacggaag gcgccgcagg agactctcaa cgggggaatc accgacatgc tcacagaact
     1681 cgcaaacttt gagaagaacg tgagccaagc tatccacaag tacaatgctt acagaaaagc
     1741 agcatctgtt atagcaaaat acccacacaa aataaagagt ggagctgaag ctaagaaatt
     1801 gcctggagta ggaacaaaaa ttgctgaaaa gattgatgag tttttagcaa ctggaaaatt
     1861 acgtaaactg gaaaagattc ggcaggatga tacgagttca tccatcaatt tcctgactcg
     1921 agttagtggc attggtccat ctgctgcaag gaagtttgta gatgaaggaa ttaaaacact
     1981 agaagatctc agaaaaaatg aagataaatt gaaccatcat cagcgaattg ggctgaaata
     2041 ttttggggac tttgaaaaaa gaattcctcg tgaagagatg ttacaaatgc aagatattgt
     2101 actaaatgaa gttaaaaaag tggattctga atacattgct acagtctgtg gcagtttcag
     2161 aagaggtgca gagtccagtg gtgacatgga tgttctcctg acccatccca gcttcacttc
     2221 agaatcaacc aaacagccaa aactgttaca tcaggttgtg gagcagttac aaaaggttca
     2281 ttttatcaca gataccctgt caaagggtga gacaaagttc atgggtgttt gccagcttcc
     2341 cagtaaaaat gatgaaaaag aatatccaca cagaagaatt gatatcaggt tgatacccaa
     2401 agatcagtat tactgtggtg ttctctattt cactgggagt gatattttca ataagaatat
     2461 gagggctcat gccctagaaa agggtttcac aatcaatgag tacaccatcc gtcccttggg
     2521 agtcattgga gttgcaggag aacccctgcc agtggatagt gaaaaagaca tctttgatta
     2581 catccagtgg aaataccggg aacccaagga ccggagcgaa tgagcggccg catcgtgact
     2641 gactgacgat ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     2701 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     2761 gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata
     2821 gcggagtgta taattcttga agacgaaagg gcctcgtgat acgcctattt ttataggtta
     2881 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     2941 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     3001 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     3061 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     3121 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     3181 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     3241 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
     3301 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     3361 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     3421 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     3481 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     3541 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa
     3601 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     3661 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     3721 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     3781 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     3841 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     3901 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     3961 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     4021 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4081 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4141 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4201 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4261 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4321 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4381 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4441 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4501 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4561 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4621 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     4681 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     4741 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     4801 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     4861 ttctccttac gcatctgtgc ggtatttcac accgcataaa ttccgacacc atcgaatggc
     4921 gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga
     4981 atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg
     5041 tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag
     5101 cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac
     5161 agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg
     5221 tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag
     5281 aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca
     5341 gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct
     5401 gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta
     5461 ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc
     5521 agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg
     5581 gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact
     5641 ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca
     5701 ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt
     5761 ccgggctgcg cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagacagct
     5821 catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca
     5881 gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc
     5941 ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcc
//
