LOCUS       40924_9401        5590 bp DNA     circular SYN 13-MAY-2021
DEFINITION  bacterial expression of the catalytic domain of murine TET1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5590)
  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 5590)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5590)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat
            Commun."; date: "2016-03-2"; pages: "
            10.1038/ncomms10806"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5590
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        115..133
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    134..158
                     /label=lac operator
                     /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             173..195
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             214..231
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      2402..2449
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2623..3411)
                     /codon_start=1
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(3412..3503)
                     /label=AmpR promoter
     rep_origin      complement(3551..4289)
                     /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."
     protein_bind    complement(4465..4486)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(4502..5581)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(join(5582..5590,1..69))
                     /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 catgggcagc agccatcacc atcatcacca cagccaggat
      241 ccgaattcga gctcggcgcg cctgcaggtc gacaagcttg cggccgcaga agctgcaccc
      301 tgtgactgtg atggaggtac acaaaaagaa aaaggcccat attatacaca ccttggggca
      361 ggaccaagtg tggctgctgt cagggagctc atggagacta ggtttggcca gaaggggaag
      421 gcaatccgga ttgagaagat agtgttcacg gggaaggaag ggaagagctc tcagggctgc
      481 ccggtcgcca agtgggtgat cagaagaagt ggtcctgaag agaagcttat ttgtttggtt
      541 cgtgagcgtg tagaccatca ctgttcgacg gctgtgatag ttgtccttat cctgctgtgg
      601 gaaggtatcc ctcgcctgat ggctgaccgc ctgtacaaag agctcactga gaacttgagg
      661 tcctacagcg gacatcccac agaccgaaga tgtaccctca acaaaaagcg tacctgcacc
      721 tgtcaaggca tcgacccaaa aacctgcgga gcgtccttct cctttggctg ttcgtggagc
      781 atgtatttca acggctgtaa gtttgggagg agtgaaaacc ccagaaaatt cagacttgct
      841 ccaaactacc ccttacataa ctactataag agaattactg gaatgagttc tgaaggaagt
      901 gacgtgaaaa ccgggtggat cattccagac cgcaagaccc tcataagcag agaggaaaaa
      961 cagcttgaaa agaatttaca agaattggct acagtattag ctccacttta caagcagatg
     1021 gctccagttg cttatcaaaa tcaggtggaa tatgaagaag ttgctggaga ctgtcgactt
     1081 ggaaatgaag aggggcgtcc tttctctggt gtcacctgtt gcatggattt ttgtgcccat
     1141 tctcacaagg acattcacaa catgcacaac ggaagcaccg tggtgtgtac gttgattcga
     1201 gcagatggcc gtgacacaaa ttgtcccgag gatgaacaac tccacgtcct gccactatac
     1261 cggcttgcag acactgatga atttggctcc gtggaaggga tgaaggccaa aatcaaatct
     1321 ggggccatcc aagtcaatgg gccaaccagg aagaggcgac tacgttttac tgagcctgtt
     1381 cctcgatgtg ggaagagggc caaaatgaag cagaaccaca ataaatcagg tactgctggt
     1441 ctcagaagga agaggatctc tgccagccca aagggagccc caggttcaca caacactaag
     1501 agcttttcat cagcctcatc tacttctcac ctagtgaaag acgaatctac agacttctgt
     1561 cccctgcagg cttcctccgc agaaacatct acctgtacgt acagtaaaac agcctcaggt
     1621 gggtttgcag aaacaagtag tattctccac tgcacaatgc cttctggagc acacagtggt
     1681 gctaatgcag ctgctgggga atgtactgga acggtgcagc ctgccgaggt ggctgctcat
     1741 cctcaccagt ctcttcccac agccgattct cccgttcatg ctgagcctct cactagtcca
     1801 tctgagcagc taacttctaa ccagtcaaac cagcagctcc ctctcctcag caattctcag
     1861 aaactggctt cctgtcaggt ggaagatgag cggcaccctg aagcggatga gcctcagcac
     1921 cccgaggacg ataacttgcc tcaacttgat gaattctggt cagacagtga ggagatctac
     1981 gccgatcctt cctttggtgg cgtggcgata gcacccattc acggctcggt gctcattgag
     2041 tgcgctcgga aggagcttca tgctaccacc tctttgcgct cccccaaacg aggggtccct
     2101 tttcgtgtgt cccttgtatt ctaccagcac aaaagcctaa acaagcctaa tcatggtttt
     2161 gatatcaaca aaattaagtg taaatgcaaa aaagtaacga aaaaaaagcc cgcagaccgg
     2221 gagtgtcctg atgtatcccc cgaagccaat ttatcacacc aaattccttc tcgagttgca
     2281 tcaaccttaa cccgagacaa tgttgttacc gtgtccccat actctctcac tcatgttgcg
     2341 ggaccctaca atcgttgggt ctaattaatt aacctaggct gctgccaccg ctgagcaata
     2401 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaacctca
     2461 ggcatttgag aagcacacgg tcacactgct tccggtagtc aataaaccgg taaaccagca
     2521 atagacataa gcggctattt aacgaccctg ccctgaaccg acgaccgggt catcgtggcc
     2581 ggatcttgcg gcccctcggc ttgaacgaat tgttagacat tatttgccga ctaccttggt
     2641 gatctcgcct ttcacgtagt ggacaaattc ttccaactga tctgcgcgcg aggccaagcg
     2701 atcttcttct tgtccaagat aagcctgtct agcttcaagt atgacgggct gatactgggc
     2761 cggcaggcgc tccattgccc agtcggcagc gacatccttc ggcgcgattt tgccggttac
     2821 tgcgctgtac caaatgcggg acaacgtaag cactacattt cgctcatcgc cagcccagtc
     2881 gggcggcgag ttccatagcg ttaaggtttc atttagcgcc tcaaatagat cctgttcagg
     2941 aaccggatca aagagttcct ccgccgctgg acctaccaag gcaacgctat gttctcttgc
     3001 ttttgtcagc aagatagcca gatcaatgtc gatcgtggct ggctcgaaga tacctgcaag
     3061 aatgtcattg cgctgccatt ctccaaattg cagttcgcgc ttagctggat aacgccacgg
     3121 aatgatgtcg tcgtgcacaa caatggtgac ttctacagcg cggagaatct cgctctctcc
     3181 aggggaagcc gaagtttcca aaaggtcgtt gatcaaagct cgccgcgttg tttcatcaag
     3241 ccttacggtc accgtaacca gcaaatcaat atcactgtgt ggcttcaggc cgccatccac
     3301 tgcggagccg tacaaatgta cggccagcaa cgtcggttcg agatggcgct cgatgacgcc
     3361 aactacctct gatagttgag tcgatacttc ggcgatcacc gcttccctca tactcttcct
     3421 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     3481 atgtatttag aaaaataaac aaatagctag ctcactcggt cgctacgctc cgggcgtgag
     3541 actgcggcgg gcgctgcgga cacatacaaa gttacccaca gattccgtgg ataagcaggg
     3601 gactaacatg tgaggcaaaa cagcagggcc gcgccggtgg cgtttttcca taggctccgc
     3661 cctcctgcca gagttcacat aaacagacgc ttttccggtg catctgtggg agccgtgagg
     3721 ctcaaccatg aatctgacag tacgggcgaa acccgacagg acttaaagat ccccaccgtt
     3781 tccggcgggt cgctccctct tgcgctctcc tgttccgacc ctgccgttta ccggatacct
     3841 gttccgcctt tctcccttac gggaagtgtg gcgctttctc atagctcaca cactggtatc
     3901 tcggctcggt gtaggtcgtt cgctccaagc tgggctgtaa gcaagaactc cccgttcagc
     3961 ccgactgctg cgccttatcc ggtaactgtt cacttgagtc caacccggaa aagcacggta
     4021 aaacgccact ggcagcagcc attggtaact gggagttcgc agaggatttg tttagctaaa
     4081 cacgcggttg ctcttgaagt gtgcgccaaa gtccggctac actggaagga cagatttggt
     4141 tgctgtgctc tgcgaaagcc agttaccacg gttaagcagt tccccaactg acttaacctt
     4201 cgatcaaacc acctccccag gtggtttttt cgtttacagg gcaaaagatt acgcgcagaa
     4261 aaaaaggatc tcaagaagat cctttgatct tttctactga accgctctag atttcagtgc
     4321 aatttatctc ttcaaatgta gcacctgaag tcagccccat acgatataag ttgtaattct
     4381 catgttagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
     4441 catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc
     4501 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4561 cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga
     4621 cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca
     4681 cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac
     4741 atgagctgtc ttcggtatcg tcgtatccca ctaccgagat gtccgcacca acgcgcagcc
     4801 cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg
     4861 cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac
     4921 tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc
     4981 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
     5041 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga
     5101 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag
     5161 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc
     5221 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc
     5281 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg
     5341 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg
     5401 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg
     5461 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg
     5521 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca
     5581 cattcaccac
//
