LOCUS       Exported                6897 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  bacterial expression of N-terminus of murine TET1.
ACCESSION   .
VERSION     .
KEYWORDS    pACYC-mTET1-N
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6897)
  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 6897)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6897
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        complement(1..78)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     124..143
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        124..142
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    143..167
                     /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             223..240
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3477..3495)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3491..3538
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3758..4417)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4332..4351
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4418..4520)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4912..4929)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5046..5590)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(5126..5145)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5815..6897)
                     /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     6859..6878
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt
       61 gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga
      121 aattaatacg actcactata ggggaattgt gagcggataa caattcccct gtagaaataa
      181 ttttgtttaa ctttaataag gagatatacc atgggcagca gccatcacca tcatcaccac
      241 agccaggatc ctatgcctct tgatgagtct gtgtatccca cagctctgat aagattcatc
      301 ttggcaggct cacagccgga tgtctttgat actaagcccc aagaaaaaac ccttataaca
      361 accccagaac aagtgggaag ccacccaaat caagttctgg atgccacctc tgtcctagga
      421 caggccttta gtactcttcc acttcagtgg ggattttctg gtgctaactt agttcaagtt
      481 gaggccctgg gcaagggttc agactcacca gaggatcttg gtgctattac tatgctaaac
      541 caacaagaaa ctgtggctat ggatatggat aggaatgcta ccccggacct gcctatcttc
      601 cttcctaagc ctccaaacac agttgctacc tacagtagcc ctcttttagg gcctgagccc
      661 cacagctcta cctcctgtgg gcttgaagtc caaggtgcta ccccgatttt aactttggac
      721 tcaggacaca ctccccaact cccaccaaac cctgagtcaa gttcagtacc tctggtaata
      781 gctgcaaacg gtacgcgagc tgagaagcag tttggtacca gccttttccc agccgtccca
      841 caagggttta cagtcgcagc cgagaatgaa gtccagcatg ccccattaga tcttacccag
      901 ggttctcagg ctgctcccag caaactggaa ggagagattt ctcgggtcag catcactggc
      961 tcagctgatg tcaaagccac agcgatgtcc atgccggtta cacaggccag tacctcttct
     1021 cccccatgta attctacgcc gccaatggtg gagaggagga agcgcaaggc ctgtggggtc
     1081 tgtgagccgt gccagcagaa ggccaactgt ggggagtgca cctactgcaa gaatcgaaag
     1141 aacagccacc agatctgtaa gaagaggaaa tgcgaggtgc tcaaaaagaa gccagaggcc
     1201 acctcacagg cacaggttac aaaagaaaac aagaggcccc agagggaaaa gaagcccaaa
     1261 gttttaaaga cagactttaa caacaaacca gtaaatggcc ccaaatcaga gtccatggac
     1321 tgcagtagac gtggtcatgg ggaagaagag caaaggttgg atttgatcac acatcccctt
     1381 gaaaatgtaa ggaagaatgc aggcggcatg acgggaattg aggtggagaa gtgggcgcca
     1441 aacaagaaat cacacttagc agagggtcaa gtcaaaggaa gctgtgatgc caatttaact
     1501 ggggtggaaa acccccagcc ctctgaagat gacaagcagc aaaccaaccc ttctccgaca
     1561 tttgcccaga ccataaggaa cggcatgaaa aatgtacact gtttacccac tgacacacac
     1621 cttccgctca acaaattgaa tcatgaggaa tttagcaagg cgttgggaaa taattcctct
     1681 aaattgctga ctgatccctc aaactgcaag gatgccatga gtgtcaccac ctccgggggg
     1741 gagtgtgatc atctcaaggg gccaaggaac accctgctct tccagaagcc tggcttgaac
     1801 tgcaggtctg gtgcagaacc caccatcttt aataatcacc ctaatacaca cagcgctggg
     1861 agtcgacctc accctcctga gaaggtaccc aacaaagaac ccaaagatgg ctctccggtt
     1921 cagccaagtc ttctatccct aatgaaggac aggagattaa cgctggaaca agtggtagcc
     1981 atagaagccc tgactcagct ctcagaagcg ccctcagaga gctcctcccc atccaagccg
     2041 gagaaggatg aagaagcgca tcagaaaacg gctagtctgc ttaacagttg caaggctatc
     2101 cttcattcgg tgagaaaaga cctccaagac ccaaacgttc aaggaaaggg ccttcaccat
     2161 gacactgttg tctttaatgg ccagaacaga acattcaagt cacctgatag tttcgctact
     2221 aaccaagcac ttataaaatc acaaggatat ccaagttcac ctacagctga gaagaaaggc
     2281 gctgctggtg ggagagctcc ctttgatggt tttgagaatt cacatccatt gcccatagaa
     2341 agtcacaatc ttgaaaactg cagtcaggta ctaagctgcg atcagaattt gagttcccat
     2401 gacccatcat gccaggatgc accctacagt caaattgaag aagatgtcgc agcacaattg
     2461 acccaacttg catccacgat taatcacatc aacgccgagg tcaggaatgc tgaaagtaca
     2521 ccagaaagcc ttgttgcaaa gaacacaaag caaaaacaca gtcaggagaa acgcatggta
     2581 catcagaagc caccctctag tacacaaact aaacccagtg taccatcagc aaagccaaag
     2641 aaggcccaga aaaaggcaag agcgacccca cacgcaaaca agcggaaaaa gaaaccccca
     2701 gcccgaagct ctcaagaaaa tgatcagaag aagcaagagc agctagctat agagtatagt
     2761 aaaatgcatg acatttggat gtcatctaag tttcaaaggt ttggccagtc tagtccacgt
     2821 agttttcctg ttctgttgag aaatattcct gtctttaacc aaatattaaa gcccgtgact
     2881 cagagtaaaa caccctcaca acataatgag ttatttcctc ctatcaatca gataaaattc
     2941 acaagaaacc ctgaattggc aaaggagaaa gtgaaggttg aaccatcaga ttctctgcca
     3001 acatgccaat tcaagacaga atccggtggg cagacatttg cagagccggc agataattct
     3061 cagggacagc ccatggtgag tgtcaatcag gaagctcacc ctctgcccca gtccccacca
     3121 tctaaccagt gtgctaatat aatggctggt gctgcccaaa cacagtttca tctaggcgct
     3181 caagagaacc tggtgcatca gataccacca ccgacgttgc ctggtacctc ccctgacaca
     3241 ctcttgccag acccagcatc aatcctcagg aaagggaaag tgttacattt cgatggaatt
     3301 acagttgtta cggagaagcg tgaagctcaa acatcaagca atggaccact gggtcccacc
     3361 acagacagtg cacagtcaga atttaaagag agcatcatgg acttacttag caagcctgca
     3421 aaaaacctga tagctggatt gaaggaacag taattaatta acctaggctg ctgccaccgc
     3481 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
     3541 gaaacctcag gcatttgaga agcacacggt cacactgctt ccggtagtca ataaaccggt
     3601 aaaccagcaa tagacataag cggctattta acgaccctgc cctgaaccga cgaccgggtc
     3661 gaatttgctt tcgaatttct gccattcatc cgcttattat cacttattca ggcgtagcac
     3721 caggcgttta agggcaccaa taactgcctt aaaaaaatta cgccccgccc tgccactcat
     3781 cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca cagacggcat
     3841 gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca
     3901 tagtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga
     3961 aactcaccca gggattggct gagacgaaaa acatattctc aataaaccct ttagggaaat
     4021 aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga aactgccgga
     4081 aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg
     4141 tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgga
     4201 actccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt
     4261 gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat
     4321 aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata
     4381 tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta gctcctgaaa
     4441 atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg
     4501 aacctcttac gtgccgatca acgtctcatt ttcgccaaaa gttggcccag ggcttcccgg
     4561 tatcaacagg gacaccagga tttatttatt ctgcgaagtg atcttccgtc acaggtattt
     4621 attcggcgca aagtgcgtcg ggtgatgctg ccaacttact gatttagtgt atgatggtgt
     4681 ttttgaggtg ctccagtggc ttctgtttct atcagctgtc cctcctgttc agctactgac
     4741 ggggtggtgc gtaacggcaa aagcaccgcc ggacatcagc gctagcggag tgtatactgg
     4801 cttactatgt tggcactgat gagggtgtca gtgaagtgct tcatgtggca ggagaaaaaa
     4861 ggctgcaccg gtgcgtcagc agaatatgtg atacaggata tattccgctt cctcgctcac
     4921 tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa atggcttacg aacggggcgg
     4981 agatttcctg gaagatgcca ggaagatact taacagggaa gtgagagggc cgcggcaaag
     5041 ccgtttttcc ataggctccg cccccctgac aagcatcacg aaatctgacg ctcaaatcag
     5101 tggtggcgaa acccgacagg actataaaga taccaggcgt ttcccctggc ggctccctcg
     5161 tgcgctctcc tgttcctgcc tttcggttta ccggtgtcat tccgctgtta tggccgcgtt
     5221 tgtctcattc cacgcctgac actcagttcc gggtaggcag ttcgctccaa gctggactgt
     5281 atgcacgaac cccccgttca gtccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5341 tccaacccgg aaagacatgc aaaagcacca ctggcagcag ccactggtaa ttgatttaga
     5401 ggagttagtc ttgaagtcat gcgccggtta aggctaaact gaaaggacaa gttttggtga
     5461 ctgcgctcct ccaagccagt tacctcggtt caaagagttg gtagctcaga gaaccttcga
     5521 aaaaccgccc tgcaaggcgg ttttttcgtt ttcagagcaa gagattacgc gcagaccaaa
     5581 acgatctcaa gaagatcatc ttattaatca gataaaatat ttctagattt cagtgcaatt
     5641 tatctcttca aatgtagcac ctgaagtcag ccccatacga tataagttgt aattctcatg
     5701 ttagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc
     5761 ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg
     5821 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     5881 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg
     5941 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct
     6001 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga
     6061 gctgtcttcg gtatcgtcgt atcccactac cgagatgtcc gcaccaacgc gcagcccgga
     6121 ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt
     6181 gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca
     6241 gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc
     6301 agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg
     6361 gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat
     6421 aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca
     6481 ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact
     6541 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc
     6601 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac
     6661 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg
     6721 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc
     6781 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac
     6841 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcac
//
