LOCUS       sequence-350545-        5993 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial vector for expression of an N-terminal GST fusion of 
            PolB(WT) with a TEV protease site located between the GST tag and 
            PolB(WT).
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(WT)
FEATURES             Location/Qualifiers
     source          1..5993
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(199..204,205..220,221..227)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    235..251
                     /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             274..927
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     885..907
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             934..951
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             952..972
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(2059..2081)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2226..2244)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2312..2416
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2417..2485,2486..3277)
                     /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(2635..2654)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3448..4036
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3937..3956
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4190..4207
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4280..4357
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4358..5440
                     /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(4377..4396)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5489..5494,5495..5512,5513..5519)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5527..5543
                     /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     5551..5567
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5551..5567
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(5563..5993,1..97)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5583..5600)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5583..5599)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5592..5614)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 tgatggcgtt ggaattagct tatcgactgc acggtgcacc aatgcttctg gcgtcaggca
       61 gccatcggaa gctgtggtat ggctgtgcag gtcgtaaatc actgcataat tcgtgtcgct
      121 caaggcgcac tcccgttctg gataatgttt tttgcgccga catcataacg gttctggcaa
      181 atattctgaa atgagctgtt gacaattaat catcggctcg tataatgtgt ggaattgtga
      241 gcggataaca atttcacaca ggaaacagta ttcatgtccc ctatactagg ttattggaaa
      301 attaagggcc ttgtgcaacc cactcgactt cttttggaat atcttgaaga aaaatatgaa
      361 gagcatttgt atgagcgcga tgaaggtgat aaatggcgaa acaaaaagtt tgaattgggt
      421 ttggagtttc ccaatcttcc ttattatatt gatggtgatg ttaaattaac acagtctatg
      481 gccatcatac gttatatagc tgacaagcac aacatgttgg gtggttgtcc aaaagagcgt
      541 gcagagattt caatgcttga aggagcggtt ttggatatta gatacggtgt ttcgagaatt
      601 gcatatagta aagactttga aactctcaaa gttgattttc ttagcaagct acctgaaatg
      661 ctgaaaatgt tcgaagatcg tttatgtcat aaaacatatt taaatggtga tcatgtaacc
      721 catcctgact tcatgttgta tgacgctctt gatgttgttt tatacatgga cccaatgtgc
      781 ctggatgcgt tcccaaaatt agtttgtttt aaaaaacgta ttgaagctat cccacaaatt
      841 gataagtact tgaaatccag caagtatata gcatggcctt tgcagggctg gcaagccacg
      901 tttggtggtg gcgaccatcc tccaaaatcg gatctggttc cgcgtggatc cgaaaacctg
      961 tattttcagg gctcgaattc ccgggtcgac atgagcaaac ggaaggcgcc gcaggagact
     1021 ctcaacgggg gaatcaccga catgctcaca gaactcgcaa actttgagaa gaacgtgagc
     1081 caagctatcc acaagtacaa tgcttacaga aaagcagcat ctgttatagc aaaataccca
     1141 cacaaaataa agagtggagc tgaagctaag aaattgcctg gagtaggaac aaaaattgct
     1201 gaaaagattg atgagttttt agcaactgga aaattacgta aactggaaaa gattcggcag
     1261 gatgatacga gttcatccat caatttcctg actcgagtta gtggcattgg tccatctgct
     1321 gcaaggaagt ttgtagatga aggaattaaa acactagaag atctcagaaa aaatgaagat
     1381 aaattgaacc atcatcagcg aattgggctg aaatattttg gggactttga aaaaagaatt
     1441 cctcgtgaag agatgttaca aatgcaagat attgtactaa atgaagttaa aaaagtggat
     1501 tctgaataca ttgctacagt ctgtggcagt ttcagaagag gtgcagagtc cagtggtgac
     1561 atggatgttc tcctgaccca tcccagcttc acttcagaat caaccaaaca gccaaaactg
     1621 ttacatcagg ttgtggagca gttacaaaag gttcatttta tcacagatac cctgtcaaag
     1681 ggtgagacaa agttcatggg tgtttgccag cttcccagta aaaatgatga aaaagaatat
     1741 ccacacagaa gaattgatat caggttgata cccaaagatc agtattactg tggtgttctc
     1801 tatttcactg ggagtgatat tttcaataag aatatgaggg ctcatgccct agaaaagggt
     1861 ttcacaatca atgagtacac catccgtccc ttgggagtca ctggagttgc aggagaaccc
     1921 ctgccagtgg atagtgaaaa agacatcttt gattacatcc agtggaaata ccgggaaccc
     1981 aaggaccgga gcgaatgagc ggccgcatcg tgactgactg acgatctgcc tcgcgcgttt
     2041 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     2101 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     2161 tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtataatt cttgaagacg
     2221 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     2281 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     2341 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     2401 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     2461 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     2521 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     2581 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     2641 tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     2701 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     2761 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     2821 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     2881 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     2941 gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga
     3001 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     3061 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     3121 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     3181 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     3241 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     3301 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     3361 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     3421 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3481 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3541 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     3601 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3661 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3721 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3781 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3841 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3901 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3961 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4021 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4081 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     4141 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     4201 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     4261 ttcacaccgc ataaattccg acaccatcga atggcgcaaa acctttcgcg gtatggcatg
     4321 atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga
     4381 tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag
     4441 ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat
     4501 tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac
     4561 ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga
     4621 tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa
     4681 agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct
     4741 ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct
     4801 tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg
     4861 actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc
     4921 attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa
     4981 tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca
     5041 aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca
     5101 gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat
     5161 ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgttaaccac
     5221 catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc
     5281 tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac
     5341 caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     5401 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtaa
     5461 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     5521 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacggat
     5581 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     5641 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     5701 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gctttgcctg gtttccggca
     5761 ccagaagcgg tgccggaaag ctggctggag tgcgatcttc ctgaggccga tactgtcgtc
     5821 gtcccctcaa actggcagat gcacggttac gatgcgccca tctacaccaa cgtgacctat
     5881 cccattacgg tcaatccgcc gtttgttccc acggagaatc cgacgggttg ttactcgctc
     5941 acatttaatg ttgatgaaag ctggctacag gaaggccaga cgcgaattat ttt
//
