LOCUS       sequence-366131-        5993 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial vector for expression of an N-terminal GST fusion of 
            PolB(K72A) with a TEV protease site located between the GST tag and 
            PolB(K72A).
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(K72A)
FEATURES             Location/Qualifiers
     source          1..5993
                     /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(3122..3144)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3289..3307)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3375..3479
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3480..3548,3549..4340)
                     /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(3698..3717)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4511..5099
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5000..5019
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5253..5270
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5343..5420
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5421..5993,1..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(5440..5459)
                     /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 gacatgagca aacggaaggc gccgcaggag actctcaacg ggggaatcac
     2101 cgacatgctc acagaactcg caaactttga gaagaacgtg agccaagcta tccacaagta
     2161 caatgcttac agaaaagcag catctgttat agcaaaatac ccacacaaaa taaagagtgg
     2221 agctgaagct aagaaattgc ctggagtagg aacaaaaatt gctgaagcga ttgatgagtt
     2281 tttagcaact ggaaaattac gtaaactgga aaagattcgg caggatgata cgagttcatc
     2341 catcaatttc ctgactcgag ttagtggcat tggtccatct gctgcaagga agtttgtaga
     2401 tgaaggaatt aaaacactag aagatctcag aaaaaatgaa gataaattga accatcatca
     2461 gcgaattggg ctgaaatatt ttggggactt tgaaaaaaga attcctcgtg aagagatgtt
     2521 acaaatgcaa gatattgtac taaatgaagt taaaaaagtg gattctgaat acattgctac
     2581 agtctgtggc agtttcagaa gaggtgcaga gtccagtggt gacatggatg ttctcctgac
     2641 ccatcccagc ttcacttcag aatcaaccaa acagccaaaa ctgttacatc aggttgtgga
     2701 gcagttacaa aaggttcatt ttatcacaga taccctgtca aagggtgaga caaagttcat
     2761 gggtgtttgc cagcttccca gtaaaaatga tgaaaaagaa tatccacaca gaagaattga
     2821 tatcaggttg atacccaaag atcagtatta ctgtggtgtt ctctatttca ctgggagtga
     2881 tattttcaat aagaatatga gggctcatgc cctagaaaag ggtttcacaa tcaatgagta
     2941 caccatccgt cccttgggag tcactggagt tgcaggagaa cccctgccag tggatagtga
     3001 aaaagacatc tttgattaca tccagtggaa ataccgggaa cccaaggacc ggagcgaatg
     3061 agcggccgca tcgtgactga ctgacgatct gcctcgcgcg tttcggtgat gacggtgaaa
     3121 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     3181 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga
     3241 cccagtcacg tagcgatagc ggagtgtata attcttgaag acgaaagggc ctcgtgatac
     3301 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     3361 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     3421 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     3481 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     3541 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     3601 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     3661 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     3721 gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     3781 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     3841 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     3901 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     3961 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     4021 ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     4081 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     4141 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     4201 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     4261 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     4321 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     4381 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     4441 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     4501 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4561 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4621 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     4681 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4741 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4801 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4861 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     4921 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     4981 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5041 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     5101 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     5161 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     5221 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     5281 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcataaatt
     5341 ccgacaccat cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga
     5401 gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg
     5461 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa
     5521 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac
     5581 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc
     5641 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg
     5701 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc
     5761 ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca
     5821 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga
     5881 cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc
     5941 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tct
//
