LOCUS       sequence-350561-        5993 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial vector for expression of an N-terminal GST fusion of 
            PolB(K206A/K244A) with a TEV protease site located between the GST 
            tag and PolB(K206A/K244A).
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(K206A/K244A)
FEATURES             Location/Qualifiers
     source          1..5993
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(4..9,10..27,28..34)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    42..58
                     /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     66..82
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     66..82
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             78..605
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(98..115)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(98..114)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(107..129)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(707..712,713..728,729..735)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    743..759
                     /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             782..1435
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1393..1415
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1442..1459
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1460..1480
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(2567..2589)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2734..2752)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2820..2924
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2925..2993,2994..3785)
                     /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(3143..3162)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3956..4544
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4445..4464
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4698..4715
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4788..4865
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4866..5948
                     /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(4885..4904)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
       61 tcacacagga aacagctatg accatgatta cggattcact ggccgtcgtt ttacaacgtc
      121 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
      181 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
      241 tgaatggcga atggcgcttt gcctggtttc cggcaccaga agcggtgccg gaaagctggc
      301 tggagtgcga tcttcctgag gccgatactg tcgtcgtccc ctcaaactgg cagatgcacg
      361 gttacgatgc gcccatctac accaacgtga cctatcccat tacggtcaat ccgccgtttg
      421 ttcccacgga gaatccgacg ggttgttact cgctcacatt taatgttgat gaaagctggc
      481 tacaggaagg ccagacgcga attatttttg atggcgttgg aattagctta tcgactgcac
      541 ggtgcaccaa tgcttctggc gtcaggcagc catcggaagc tgtggtatgg ctgtgcaggt
      601 cgtaaatcac tgcataattc gtgtcgctca aggcgcactc ccgttctgga taatgttttt
      661 tgcgccgaca tcataacggt tctggcaaat attctgaaat gagctgttga caattaatca
      721 tcggctcgta taatgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagtatt
      781 catgtcccct atactaggtt attggaaaat taagggcctt gtgcaaccca ctcgacttct
      841 tttggaatat cttgaagaaa aatatgaaga gcatttgtat gagcgcgatg aaggtgataa
      901 atggcgaaac aaaaagtttg aattgggttt ggagtttccc aatcttcctt attatattga
      961 tggtgatgtt aaattaacac agtctatggc catcatacgt tatatagctg acaagcacaa
     1021 catgttgggt ggttgtccaa aagagcgtgc agagatttca atgcttgaag gagcggtttt
     1081 ggatattaga tacggtgttt cgagaattgc atatagtaaa gactttgaaa ctctcaaagt
     1141 tgattttctt agcaagctac ctgaaatgct gaaaatgttc gaagatcgtt tatgtcataa
     1201 aacatattta aatggtgatc atgtaaccca tcctgacttc atgttgtatg acgctcttga
     1261 tgttgtttta tacatggacc caatgtgcct ggatgcgttc ccaaaattag tttgttttaa
     1321 aaaacgtatt gaagctatcc cacaaattga taagtacttg aaatccagca agtatatagc
     1381 atggcctttg cagggctggc aagccacgtt tggtggtggc gaccatcctc caaaatcgga
     1441 tctggttccg cgtggatccg aaaacctgta ttttcagggc tcgaattccc gggtcgacat
     1501 gagcaaacgg aaggcgccgc aggagactct caacggggga atcaccgaca tgctcacaga
     1561 actcgcaaac tttgagaaga acgtgagcca agctatccac aagtacaatg cttacagaaa
     1621 agcagcatct gttatagcaa aatacccaca caaaataaag agtggagctg aagctaagaa
     1681 attgcctgga gtaggaacaa aaattgctga aaagattgat gagtttttag caactggaaa
     1741 attacgtaaa ctggaaaaga ttcggcagga tgatacgagt tcatccatca atttcctgac
     1801 tcgagttagt ggcattggtc catctgctgc aaggaagttt gtagatgaag gaattaaaac
     1861 actagaagat ctcagaaaaa atgaagataa attgaaccat catcagcgaa ttgggctgaa
     1921 atattttggg gactttgaaa aaagaattcc tcgtgaagag atgttacaaa tgcaagatat
     1981 tgtactaaat gaagttaaaa aagtggattc tgaatacatt gctacagtct gtggcagttt
     2041 cagaagaggt gcagagtcca gtggtgacat ggatgttctc ctgacccatc ccagcttcac
     2101 ttcagaatca accgcacagc caaaactgtt acatcaggtt gtggagcagt tacaaaaggt
     2161 tcattttatc acagataccc tgtcaaaggg tgagacaaag ttcatgggtg tttgccagct
     2221 tcccagtgca aatgatgaaa aagaatatcc acacagaaga attgatatca ggttgatacc
     2281 caaagatcag tattactgtg gtgttctcta tttcactggg agtgatattt tcaataagaa
     2341 tatgagggct catgccctag aaaagggttt cacaatcaat gagtacacca tccgtccctt
     2401 gggagtcact ggagttgcag gagaacccct gccagtggat agtgaaaaag acatctttga
     2461 ttacatccag tggaaatacc gggaacccaa ggaccggagc gaatgagcgg ccgcatcgtg
     2521 actgactgac gatctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     2581 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     2641 agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg
     2701 atagcggagt gtataattct tgaagacgaa agggcctcgt gatacgccta tttttatagg
     2761 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     2821 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     2881 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     2941 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     3001 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     3061 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     3121 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc
     3181 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     3241 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     3301 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     3361 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     3421 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa
     3481 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     3541 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     3601 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     3661 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     3721 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     3781 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     3841 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     3901 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     3961 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     4021 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     4081 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     4141 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     4201 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     4261 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     4321 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     4381 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     4441 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     4501 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     4561 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     4621 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     4681 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
     4741 attttctcct tacgcatctg tgcggtattt cacaccgcat aaattccgac accatcgaat
     4801 ggcgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg
     4861 tgaatgtgaa accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga
     4921 ccgtttcccg cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg
     4981 aagcggcgat ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca
     5041 aacagtcgtt gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa
     5101 ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg
     5161 tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg
     5221 tcagtgggct gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg
     5281 cctgcactaa tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta
     5341 ttattttctc ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc
     5401 accagcaaat cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg
     5461 ctggctggca taaatatctc actcgcaatc aaattcagcc gatagcggaa cgggaaggcg
     5521 actggagtgc catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc
     5581 ccactgcgat gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg
     5641 agtccgggct gcgcgttggt gcggatatct cggtagtggg atacgacgat accgaagaca
     5701 gctcatgtta tatcccgccg ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa
     5761 ccagcgtgga ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt
     5821 tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc caatacgcaa accgcctctc
     5881 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     5941 ggcagtgagc gcaacgcaat taatgtaagt tagctcactc attaggcacc cca
//
