LOCUS       sequence-126679-        6553 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of human TDG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6553)
  AUTHORS   Schuermann D, Scheidegger SP, Weber AR, Bjoras M, Leumann CJ, Schar 
            P
  TITLE     3CAPS - a structural AP-site analogue as a tool to investigate DNA 
            base excision repair.
  JOURNAL   Nucleic Acids Res. 2016 Jan 4. pii: gkv1520.
  PUBMED    26733580
REFERENCE   2  (bases 1 to 6553)
  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"; journalName: "Nucleic 
            Acids Res. 2016 Jan 4. pii: gkv1520."
COMMENT     Sequence Label: pET28-hTDG
FEATURES             Location/Qualifiers
     source          1..6553
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(4..81)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     287..306
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     390..409
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        390..408
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    409..433
                     /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)."
     RBS             448..470
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             489..506
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             516..533
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1803..1820
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1873..1891)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1887..1934
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1971..2426
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2058..2077)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2268..2289
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2519..3334)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2519..3334)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3242..3261
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3326..3345
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3456..4044
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3945..3964
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4198..4215
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4230..4372
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4298..4317)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4458..4480
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4474..4665)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5474..6553,1..3))
                     /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     6518..6537
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt
       61 tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta
      121 ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat
      181 gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga
      241 aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga
      301 tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt
      361 agaggatcga gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc
      421 ggataacaat tcccctctag aaataatttt gtttaacttt aagaaggaga tataccatgg
      481 gcagcagcca tcatcatcat catcacagca gcggcctggt gccgcgcggc agccatatgg
      541 ctagcgaagc ggagaacgcg ggcagctatt cccttcagca agctcaagct ttttatacgt
      601 ttccatttca acaactgatg gctgaagctc ctaatatggc agttgtgaat gaacagcaaa
      661 tgccagaaga agttccagcc ccagctcctg ctcaggaacc agtgcaagag gctccaaaag
      721 gaagaaaaag aaaacccaga acaacagaac caaaacaacc agtggaaccc aaaaaacctg
      781 ttgagtcaaa aaaatctggc aagtctgcaa aatcaaaaga aaaacaagaa aaaattacag
      841 acacatttaa agtaaaaaga aaagtagacc gttttaatgg tgtttcagaa gctgaacttc
      901 tgaccaagac tctccccgat attttgacct tcaatctgga cattgtcatt attggcataa
      961 acccgggact aatggctgct tacaaagggc atcattaccc tggacctgga aaccattttt
     1021 ggaagtgttt gtttatgtca gggctcagtg aggtccagct gaaccatatg gatgatcaca
     1081 ctctaccagg gaagtatggt attggattta ccaacatggt ggaaaggacc acgcccggca
     1141 gcaaagatct ctccagtaaa gaatttcgtg aaggaggacg tattctagta cagaaattac
     1201 agaaatatca gccacgaata gcagtgttta atggaaaatg tatttatgaa atttttagta
     1261 aagaagtttt tggagtaaag gttaagaact tggaatttgg gcttcagccc cataagattc
     1321 cagacacaga aactctctgc tatgttatgc catcatccag tgcaagatgt gctcagtttc
     1381 ctcgagccca agacaaagtt cattactaca taaaactgaa ggacttaaga gatcagttga
     1441 aaggcattga acgaaatatg gacgttcaag aggtgcaata tacatttgac ctacagcttg
     1501 cccaagagga tgcaaagaag atggctgtta aggaagaaaa atatgatcca ggttatgagg
     1561 cagcatatgg tggtgcttac ggagaaaatc catgcagcag tgaaccttgt ggcttctctt
     1621 caaatgggct aattgagagc gtggagttaa gaggagaatc agctttcagt ggcattccta
     1681 atgggcagtg gatgacccag tcatttacag accaaattcc ttcctttagt aatcactgtg
     1741 gaacacaaga acaggaagaa gaaagccatg cttaagtcga caagcttgcg gccgcactcg
     1801 agcaccacca ccaccaccac tgagatccgg ctgctaacaa agcccgaaag gaagctgagt
     1861 tggctgctgc caccgctgag caataactag cataacccct tggggcctct aaacgggtct
     1921 tgaggggttt tttgctgaaa ggaggaacta tatccggatt ggcgaatggg acgcgccctg
     1981 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2041 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2101 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     2161 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     2221 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     2281 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     2341 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     2401 taacaaaata ttaacgttta caatttcagg tggcactttt cggggaaatg tgcgcggaac
     2461 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga attaattctt
     2521 agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac
     2581 catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata
     2641 ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta
     2701 ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg
     2761 aatccggtga gaatggcaaa agtttatgca tttctttcca gacttgttca acaggccagc
     2821 cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg
     2881 cctgagcgag acgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat
     2941 gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt
     3001 cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat
     3061 caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta
     3121 gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca
     3181 actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat
     3241 tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc
     3301 tagagcaaga cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt
     3361 aagcagacag ttttattgtt catgaccaaa atcccttaac gtgagttttc gttccactga
     3421 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     3481 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     3541 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     3601 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     3661 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     3721 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     3781 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     3841 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     3901 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     3961 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     4021 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     4081 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     4141 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     4201 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
     4261 tgcggtattt cacaccgcat atatggtgca ctctcagtac aatctgctct gatgccgcat
     4321 agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca
     4381 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     4441 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     4501 acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc
     4561 tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct
     4621 gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta
     4681 agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat
     4741 acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc
     4801 ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa
     4861 tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat
     4921 aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac
     4981 cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc
     5041 gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct
     5101 caacgacagg agcacgatca tgcgcacccg tggggccgcc atgccggcga taatggcctg
     5161 cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa
     5221 gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc
     5281 gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac
     5341 agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt
     5401 gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt
     5461 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     5521 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc
     5581 ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt
     5641 gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta
     5701 acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatatccg
     5761 caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt
     5821 tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa
     5881 aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag
     5941 tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg
     6001 ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac
     6061 cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata
     6121 acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat
     6181 agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg
     6241 cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc
     6301 gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa
     6361 cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat
     6421 tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct
     6481 ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata
     6541 acgttactgg ttt
//
