LOCUS       sequence-220964-        7818 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of catalytically dead murine TDG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7818)
  AUTHORS   Weber AR, Krawczyk C, Robertson AB, Kusnierczyk A, Vagbo CB, 
            Schuermann D, Klungland A, Schar P
  TITLE     Biochemical reconstitution of TET1-TDG-BER-dependent active DNA 
            demethylation reveals a highly coordinated mechanism.
  JOURNAL   Nat Commun. 2016 Mar 2;7:10806. doi: 10.1038/ncomms10806.
  PUBMED    26932196
REFERENCE   2  (bases 1 to 7818)
  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: "Nat 
            Commun."; date: "2016-03-2"; volume: "7:10806. doi"; pages: " 
            10.1038/ncomms10806"
COMMENT     Sequence Label: pTG-mTDGa.1
FEATURES             Location/Qualifiers
     source          1..7818
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1358..1381
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1391..2047
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     complement(2096..2114)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2110..2157
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2217..2321
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2322..2390,2391..3182)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2540..2559)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3224..3737)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3419..3440)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3631..3650
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      3848..4436
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4337..4356
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4590..4607
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(4629..4648)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4789..4811
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4805..4996)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5553..6635)
                     /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     6597..6616
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6636..6713)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      6866..6952
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7049..7135
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7232..7318
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7415..7501
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7598..7684
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataaccat ggacgcagag gccgcgcgca gctattctct
      121 ggagcaagtt caagctttgt attcatttcc atttcaacaa atgatggcag aagttcctaa
      181 catggcagtc acgactggac agcaggtgcc agcagtagct cctaacatgg caaccgtgac
      241 agaacagcag gtgcccgcag acgctcctgt ccaggaacct gcaccagaag ctccaaagag
      301 aaggaaaagg aaacccagag cagcagagcc ccaggaacca gtggagccca aaaaacctgc
      361 tacgtcgaag aaatccggca agtctacaaa atcaaaggaa aagcaggaga aaatcacaga
      421 cgcgtttaaa gtgaaaagga aagtggaccg cttcaacggc gtctctgaag ctgagcttct
      481 gaccaagact cttcctgaca ttttgacctt caatctggat attgtgatca ttggcattgc
      541 cccgggatta atggctgctt acaaaggaca tcactaccct gggcctggaa atcacttctg
      601 gaagtgtctg ttcatgtcgg ggctgagtga ggtgcagctg aatcacatgg atgaccacac
      661 cttacccggc aagtacggca tcggattcac caacatggtg gaacggacga cgccgggcag
      721 caaggatctg tctagtaaag agttccggga aggagggcgc atcctggtgc agaaactgca
      781 gaaatatcag ccacgaatag cggtgtttaa tggaaaatgt atttatgaaa ttttcagtaa
      841 agaagttttt ggagtaaagg ttaagaactt ggaatttggg cttcaacccc acaagatccc
      901 agacacagaa actctgtgct acgtcatgcc gtcgtccagc gccagatgtg ctcagtttcc
      961 ccgggcccag gacaaagttc attactacat taagctgaag gacttgagag accagctgaa
     1021 aggcattgaa cgcaacaccg acgttcagga agtgcagtat acatttgacc tgcagcttgc
     1081 gcaagaggac gcaaagaaga tggctgttaa ggaagaaaag tatgatccag gctatgaggc
     1141 agcttacggc ggcgcctatg gggaaaaccc atgtaatggg gaaccttgtg gcattgcttc
     1201 aaatgggcta acagcccaca gtgcggagcc gagaggagaa gcgacccccg gcgatgttcc
     1261 aaatgggcag tggatggcac agtcgtttgc agagcagatc ccttctttta ataattgtgg
     1321 gacccgagag caggaagaag agagccacgc tctcgagctg gaagttctgt tccaggggcc
     1381 cggatccgga atgtccccta tactaggtta ttggaaaatt aagggccttg tgcaacccac
     1441 tcgacttctt ttggaatatc ttgaagaaaa atatgaagag catttgtatg agcgcgatga
     1501 aggtgataaa tggcgaaaca aaaagtttga attgggtttg gagtttccca atcttcctta
     1561 ttatattgat ggtgatgtta aattaacaca gtctatggcc atcatacgtt atatagctga
     1621 caagcacaac atgttgggtg gttgtccaaa agagcgtgca gagatttcaa tgcttgaagg
     1681 agcggttttg gatattagat acggtgtttc gagaattgca tatagtaaag actttgaaac
     1741 tctcaaagtt gattttctta gcaagctacc tgaaatgctg aaaatgttcg aagatcgttt
     1801 atgtcataaa acatatttaa atggtgatca tgtaacccat cctgacttca tgttgtatga
     1861 cgctcttgat gttgttttat acatggaccc aatgtgcctg gatgcgttcc caaaattagt
     1921 ttgttttaaa aaacgtattg aagctatccc acaaattgat aagtacttga aatccagcaa
     1981 gtatatagca tggcctttgc agggctggca agccacgttt ggtggtggcg accatcctcc
     2041 aaaatagatc cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct
     2101 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg
     2161 aaaggaggaa ctatatccgg ataactacgt caggtggcac ttttcgggga aatgtgcgcg
     2221 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     2281 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     2341 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     2401 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     2461 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt ttcccaatga
     2521 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
     2581 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     2641 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     2701 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     2761 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     2821 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     2881 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     2941 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     3001 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     3061 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     3121 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     3181 aactgtcaga ccaagtttac tcatatatac tttagattga tttaccccgg ttgataatca
     3241 gaaaagcccc aaaaacagga agattgtata agcaaatatt taaattgtaa acgttaatat
     3301 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     3361 aatcggcaaa atcccttata aatcaaaaga atagcccgag atagggttga gtgttgttcc
     3421 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     3481 cgtctatcag ggcgatggcc cactacgtga accatcaccc aaatcaagtt ttttggggtc
     3541 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     3601 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     3661 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     3721 gccgctacag ggcgcgtaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     3781 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3841 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3901 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3961 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     4021 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4081 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4141 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4201 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4261 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4321 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4381 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4441 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4501 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     4561 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gctatggtgc
     4621 actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc
     4681 tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac
     4741 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     4801 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat
     4861 cagcgtggtc gtgcagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga
     4921 gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt
     4981 tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga
     5041 taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt
     5101 tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa
     5161 tcactcaggg tcaatgccag ccgaacgcca gcaagacgta gcccagcgcg tcggccgcca
     5221 tgccggcgat aatggcctgc ttctcgccga aacgtttggt ggcgggacca gtgacgaagg
     5281 cttgagcgag ggcgtgcaag attccgaata ccgcaagcga caggccgatc atcgtcgcgc
     5341 tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc tgtcctacga
     5401 gttgcatgat aaagaagaca gtcataagtg cggcgacgat agtcatgccc cgcgcccacc
     5461 ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat
     5521 gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     5581 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5641 ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc
     5701 gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa
     5761 tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat
     5821 cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg
     5881 cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc
     5941 atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc
     6001 ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag
     6061 acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc
     6121 tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg
     6181 tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca
     6241 tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg
     6301 tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg
     6361 gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg
     6421 gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc
     6481 acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc
     6541 gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca
     6601 tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct
     6661 tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt gtccgggatc
     6721 tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc
     6781 gttgagcacc gccgccgcaa ggaatggtgc atgccggcat gccgcccttt cgtcttcaag
     6841 aattaattcc caattcccca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
     6901 ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg
     6961 agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata
     7021 aactgccagg aattaattcc ccaggcatca aataaaacga aaggctcagt cgaaagactg
     7081 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc
     7141 gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc
     7201 ataaactgcc aggaattaat tccccaggca tcaaataaaa cgaaaggctc agtcgaaaga
     7261 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
     7321 gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc
     7381 gccataaact gccaggaatt aattccccag gcatcaaata aaacgaaagg ctcagtcgaa
     7441 agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa
     7501 tccgccggga gcggatttga acgttgcgaa gcaacggccc ggagggtggc gggcaggacg
     7561 cccgccataa actgccagga attaattccc caggcatcaa ataaaacgaa aggctcagtc
     7621 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac
     7681 aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg
     7741 acgcccgcca taaactgcca ggaattgggg atcggaatta attcccggtt taaaccgggg
     7801 atctcgatcc cgcgaaat
//
