LOCUS       sequence-153729-        9726 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of murine TET1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9726)
  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 9726)
  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: pCDF-mTET1
FEATURES             Location/Qualifiers
     source          1..9726
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     115..134
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        115..133
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    134..158
                     /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             6409..6426
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             6442..6486
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(6524..6542)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6538..6585
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(6756..7547)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(7548..7639)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7687..8425)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     CDS             complement(8635..9717)
                     /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     9679..9698
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(join(9718..9726,1..69))
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
       61 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aaattaatac
      121 gactcactat aggggaattg tgagcggata acaattcccc tgtagaaata attttgttta
      181 actttaataa ggagatatac catggacgcg tgcggccgca tcatgtctcg gtcccgcccc
      241 gcaaagcctt ccaaatcagt caaaacaaag ctacagaaaa agaaagacat ccagatgaag
      301 acgaagacat caaagcaggc tgtcagacat ggggcatcag cgaaggcagt gaaccccgga
      361 aaaccgaagc aattaattaa aagaagagat ggtaagaagg aaacggaaga caaaaccccg
      421 acaccggcgc cgagttttct gacgcgagcc ggggccgccc gaatgaatcg ggatcgaaac
      481 caagttcttt ttcagaaccc agactcctta acttgcaacg ggtttacaat ggctcttcga
      541 agaacctctc ttagctggcg actctcccag cgcccagtag tcacacccaa acctaagaaa
      601 gtaccacctt caaagaagca gtgtacacat aatatccagg acgagccggg ggtaaagcac
      661 tctgaaaatg attcagtccc cagccaacac gctaccgtgt ccccaggcac agagaatggt
      721 gaacagaaca ggtgtttagt tgaaggggag agccaagaga taacccagtc ttgccctgta
      781 tttgaggaaa gaatagaaga tacccaaagc tgtatttccg catctgggaa cctggaagca
      841 gaaatttctt ggccactgga agggacacat tgtgaagaac tactttctca tcagacatct
      901 gataatgagt gcacttcccc tcaagagtgt gctccattgc cccaaagatc aacttccgaa
      961 gttacctctc agaaaaacac tagcaatcag ttagcagact tgagctcaca agtagagtct
     1021 atcaagctgt ctgatccttc tccgaatcct acgggaagcg accataatgg ctttccagac
     1081 tccagtttta ggatcgttcc tgaattggac ctgaaaacct gcatgcctct tgatgagtct
     1141 gtgtatccca cagctctgat aagattcatc ttggcaggct cacagccgga tgtctttgat
     1201 actaagcccc aagaaaaaac ccttataaca accccagaac aagtgggaag ccacccaaat
     1261 caagttctgg atgccacctc tgtcctagga caggccttta gtactcttcc acttcagtgg
     1321 ggattttctg gtgctaactt agttcaagtt gaggccctgg gcaagggttc agactcacca
     1381 gaggatcttg gtgctattac tatgctaaac caacaagaaa ctgtggctat ggatatggat
     1441 aggaatgcta ccccggacct gcctatcttc cttcctaagc ctccaaacac agttgctacc
     1501 tacagtagcc ctcttttagg gcctgagccc cacagctcta cctcctgtgg gcttgaagtc
     1561 caaggtgcta ccccgatttt aactttggac tcaggacaca ctccccaact cccaccaaac
     1621 cctgagtcaa gttcagtacc tctggtaata gctgcaaacg gtacgcgagc tgagaagcag
     1681 tttggtacca gccttttccc agccgtccca caagggttta cagtcgcagc cgagaatgaa
     1741 gtccagcatg ccccattaga tcttacccag ggttctcagg ctgctcccag caaactggaa
     1801 ggagagattt ctcgggtcag catcactggc tcagctgatg tcaaagccac agcgatgtcc
     1861 atgccggtta cacaggccag tacctcttct cccccatgta attctacgcc gccaatggtg
     1921 gagaggagga agcgcaaggc ctgtggggtc tgtgagccgt gccagcagaa ggccaactgt
     1981 ggggagtgca cctactgcaa gaatcgaaag aacagccacc agatctgtaa gaagaggaaa
     2041 tgcgaggtgc tcaaaaagaa gccagaggcc acctcacagg cacaggttac aaaagaaaac
     2101 aagaggcccc agagggaaaa gaagcccaaa gttttaaaga cagactttaa caacaaacca
     2161 gtaaatggcc ccaaatcaga gtccatggac tgcagtagac gtggtcatgg ggaagaagag
     2221 caaaggttgg atttgatcac acatcccctt gaaaatgtaa ggaagaatgc aggcggcatg
     2281 acgggaattg aggtggagaa gtgggcgcca aacaagaaat cacacttagc agagggtcaa
     2341 gtcaaaggaa gctgtgatgc caatttaact ggggtggaaa acccccagcc ctctgaagat
     2401 gacaagcagc aaaccaaccc ttctccgaca tttgcccaga ccataaggaa cggcatgaaa
     2461 aatgtacact gtttacccac tgacacacac cttccgctca acaaattgaa tcatgaggaa
     2521 tttagcaagg cgttgggaaa taattcctct aaattgctga ctgatccctc aaactgcaag
     2581 gatgccatga gtgtcaccac ctccgggggg gagtgtgatc atctcaaggg gccaaggaac
     2641 accctgctct tccagaagcc tggcttgaac tgcaggtctg gtgcagaacc caccatcttt
     2701 aataatcacc ctaatacaca cagcgctggg agtcgacctc accctcctga gaaggtaccc
     2761 aacaaagaac ccaaagatgg ctctccggtt cagccaagtc ttctatccct aatgaaggac
     2821 aggagattaa cgctggaaca agtggtagcc atagaagccc tgactcagct ctcagaagcg
     2881 ccctcagaga gctcctcccc atccaagccg gagaaggatg aagaagcgca tcagaaaacg
     2941 gctagtctgc ttaacagttg caaggctatc cttcattcgg tgagaaaaga cctccaagac
     3001 ccaaacgttc aaggaaaggg ccttcaccat gacactgttg tctttaatgg ccagaacaga
     3061 acattcaagt cacctgatag tttcgctact aaccaagcac ttataaaatc acaaggatat
     3121 ccaagttcac ctacagctga gaagaaaggc gctgctggtg ggagagctcc ctttgatggt
     3181 tttgagaatt cacatccatt gcccatagaa agtcacaatc ttgaaaactg cagtcaggta
     3241 ctaagctgcg atcagaattt gagttcccat gacccatcat gccaggatgc accctacagt
     3301 caaattgaag aagatgtcgc agcacaattg acccaacttg catccacgat taatcacatc
     3361 aacgccgagg tcaggaatgc tgaaagtaca ccagaaagcc ttgttgcaaa gaacacaaag
     3421 caaaaacaca gtcaggagaa acgcatggta catcagaagc caccctctag tacacaaact
     3481 aaacccagtg taccatcagc aaagccaaag aaggcccaga aaaaggcaag agcgacccca
     3541 cacgcaaaca agcggaaaaa gaaaccccca gcccgaagct ctcaagaaaa tgatcagaag
     3601 aagcaagagc agctagctat agagtatagt aaaatgcatg acatttggat gtcatctaag
     3661 tttcaaaggt ttggccagtc tagtccacgt agttttcctg ttctgttgag aaatattcct
     3721 gtctttaacc aaatattaaa gcccgtgact cagagtaaaa caccctcaca acataatgag
     3781 ttatttcctc ctatcaatca gataaaattc acaagaaacc ctgaattggc aaaggagaaa
     3841 gtgaaggttg aaccatcaga ttctctgcca acatgccaat tcaagacaga atccggtggg
     3901 cagacatttg cagagccggc agataattct cagggacagc ccatggtgag tgtcaatcag
     3961 gaagctcacc ctctgcccca gtccccacca tctaaccagt gtgctaatat aatggctggt
     4021 gctgcccaaa cacagtttca tctaggcgct caagagaacc tggtgcatca gataccacca
     4081 ccgacgttgc ctggtacctc ccctgacaca ctcttgccag acccagcatc aatcctcagg
     4141 aaagggaaag tgttacattt cgatggaatt acagttgtta cggagaagcg tgaagctcaa
     4201 acatcaagca atggaccact gggtcccacc acagacagtg cacagtcaga atttaaagag
     4261 agcatcatgg acttacttag caagcctgca aaaaacctga tagctggatt gaaggaacag
     4321 gaagctgcac cctgtgactg tgatggaggt acacaaaaag aaaaaggccc atattataca
     4381 caccttgggg caggaccaag tgtggctgct gtcagggagc tcatggagac taggtttggc
     4441 cagaagggga aggcaatccg gattgagaag atagtgttca cggggaagga agggaagagc
     4501 tctcagggct gcccggtcgc caagtgggtg atcagaagaa gtggtcctga agagaagctt
     4561 atttgtttgg ttcgtgagcg tgtagaccat cactgttcga cggctgtgat agttgtcctt
     4621 atcctgctgt gggaaggtat ccctcgcctg atggctgacc gcctgtacaa agagctcact
     4681 gagaacttga ggtcctacag cggacatccc acagaccgaa gatgtaccct caacaaaaag
     4741 cgtacctgca cctgtcaagg catcgaccca aaaacctgcg gagcgtcctt ctcctttggc
     4801 tgttcgtgga gcatgtattt caacggctgt aagtttggga ggagtgaaaa ccccagaaaa
     4861 ttcagacttg ctccaaacta ccccttacat aactactata agagaattac tggaatgagt
     4921 tctgaaggaa gtgacgtgaa aaccgggtgg atcattccag accgcaagac cctcataagc
     4981 agagaggaaa aacagcttga aaagaattta caagaattgg ctacagtatt agctccactt
     5041 tacaagcaga tggctccagt tgcttatcaa aatcaggtgg aatatgaaga agttgctgga
     5101 gactgtcgac ttggaaatga agaggggcgt cctttctctg gtgtcacctg ttgcatggat
     5161 ttttgtgccc attctcacaa ggacattcac aacatgcaca acggaagcac cgtggtgtgt
     5221 acgttgattc gagcagatgg ccgtgacaca aattgtcccg aggatgaaca actccacgtc
     5281 ctgccactat accggcttgc agacactgat gaatttggct ccgtggaagg gatgaaggcc
     5341 aaaatcaaat ctggggccat ccaagtcaat gggccaacca ggaagaggcg actacgtttt
     5401 actgagcctg ttcctcgatg tgggaagagg gccaaaatga agcagaacca caataaatca
     5461 ggtactgctg gtctcagaag gaagaggatc tctgccagcc caaagggagc cccaggttca
     5521 cacaacacta agagcttttc atcagcctca tctacttctc acctagtgaa agacgaatct
     5581 acagacttct gtcccctgca ggcttcctcc gcagaaacat ctacctgtac gtacagtaaa
     5641 acagcctcag gtgggtttgc agaaacaagt agtattctcc actgcacaat gccttctgga
     5701 gcacacagtg gtgctaatgc agctgctggg gaatgtactg gaacggtgca gcctgccgag
     5761 gtggctgctc atcctcacca gtctcttccc acagccgatt ctcccgttca tgctgagcct
     5821 ctcactagtc catctgagca gctaacttct aaccagtcaa accagcagct ccctctcctc
     5881 agcaattctc agaaactggc ttcctgtcag gtggaagatg agcggcaccc tgaagcggat
     5941 gagcctcagc accccgagga cgataacttg cctcaacttg atgaattctg gtcagacagt
     6001 gaggagatct acgccgatcc ttcctttggt ggcgtggcga tagcacccat tcacggctcg
     6061 gtgctcattg agtgcgctcg gaaggagctt catgctacca cctctttgcg ctcccccaaa
     6121 cgaggggtcc cttttcgtgt gtcccttgta ttctaccagc acaaaagcct aaacaagcct
     6181 aatcatggtt ttgatatcaa caaaattaag tgtaaatgca aaaaagtaac gaaaaaaaag
     6241 cccgcagacc gggagtgtcc tgatgtatcc cccgaagcca atttatcaca ccaaattcct
     6301 tctcgagttg catcaacctt aacccgagac aatgttgtta ccgtgtcccc atactctctc
     6361 actcatgttg cgggacccta caatcgttgg gtctatctag ttcccgggca tcatcaccat
     6421 caccattgac tcgagtctgg taaagaaacc gctgctgcga aatttgaacg ccagcacatg
     6481 gactcgtcta ctagcgcagc ttaattaacc taggctgctg ccaccgctga gcaataacta
     6541 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa acctcaggca
     6601 tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa ccagcaatag
     6661 acataagcgg ctatttaacg accctgccct gaaccgacga ccgggtcatc gtggccggat
     6721 cttgcggccc ctcggcttga acgaattgtt agacattatt tgccgactac cttggtgatc
     6781 tcgcctttca cgtagtggac aaattcttcc aactgatctg cgcgcgaggc caagcgatct
     6841 tcttcttgtc caagataagc ctgtctagct tcaagtatga cgggctgata ctgggccggc
     6901 aggcgctcca ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc ggttactgcg
     6961 ctgtaccaaa tgcgggacaa cgtaagcact acatttcgct catcgccagc ccagtcgggc
     7021 ggcgagttcc atagcgttaa ggtttcattt agcgcctcaa atagatcctg ttcaggaacc
     7081 ggatcaaaga gttcctccgc cgctggacct accaaggcaa cgctatgttc tcttgctttt
     7141 gtcagcaaga tagccagatc aatgtcgatc gtggctggct cgaagatacc tgcaagaatg
     7201 tcattgcgct gccattctcc aaattgcagt tcgcgcttag ctggataacg ccacggaatg
     7261 atgtcgtcgt gcacaacaat ggtgacttct acagcgcgga gaatctcgct ctctccaggg
     7321 gaagccgaag tttccaaaag gtcgttgatc aaagctcgcc gcgttgtttc atcaagcctt
     7381 acggtcaccg taaccagcaa atcaatatca ctgtgtggct tcaggccgcc atccactgcg
     7441 gagccgtaca aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat gacgccaact
     7501 acctctgata gttgagtcga tacttcggcg atcaccgctt ccctcatact cttccttttt
     7561 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7621 atttagaaaa ataaacaaat agctagctca ctcggtcgct acgctccggg cgtgagactg
     7681 cggcgggcgc tgcggacaca tacaaagtta cccacagatt ccgtggataa gcaggggact
     7741 aacatgtgag gcaaaacagc agggccgcgc cggtggcgtt tttccatagg ctccgccctc
     7801 ctgccagagt tcacataaac agacgctttt ccggtgcatc tgtgggagcc gtgaggctca
     7861 accatgaatc tgacagtacg ggcgaaaccc gacaggactt aaagatcccc accgtttccg
     7921 gcgggtcgct ccctcttgcg ctctcctgtt ccgaccctgc cgtttaccgg atacctgttc
     7981 cgcctttctc ccttacggga agtgtggcgc tttctcatag ctcacacact ggtatctcgg
     8041 ctcggtgtag gtcgttcgct ccaagctggg ctgtaagcaa gaactccccg ttcagcccga
     8101 ctgctgcgcc ttatccggta actgttcact tgagtccaac ccggaaaagc acggtaaaac
     8161 gccactggca gcagccattg gtaactggga gttcgcagag gatttgttta gctaaacacg
     8221 cggttgctct tgaagtgtgc gccaaagtcc ggctacactg gaaggacaga tttggttgct
     8281 gtgctctgcg aaagccagtt accacggtta agcagttccc caactgactt aaccttcgat
     8341 caaaccacct ccccaggtgg ttttttcgtt tacagggcaa aagattacgc gcagaaaaaa
     8401 aggatctcaa gaagatcctt tgatcttttc tactgaaccg ctctagattt cagtgcaatt
     8461 tatctcttca aatgtagcac ctgaagtcag ccccatacga tataagttgt aattctcatg
     8521 ttagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc
     8581 ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg
     8641 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     8701 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg
     8761 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct
     8821 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga
     8881 gctgtcttcg gtatcgtcgt atcccactac cgagatgtcc gcaccaacgc gcagcccgga
     8941 ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt
     9001 gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca
     9061 gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc
     9121 agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg
     9181 gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat
     9241 aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca
     9301 ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact
     9361 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc
     9421 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac
     9481 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg
     9541 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc
     9601 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac
     9661 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt
     9721 caccac
//
