LOCUS       sequence-442713-        8102 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express flag-tagged mouse Tet1 catalytic domain..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8102)
  AUTHORS   Ito S, Shen L, Dai Q, Wu SC, Collins LB, Swenberg JA, He C, Zhang Y
  TITLE     Tet proteins can convert 5-methylcytosine to 5-formylcytosine and 
            5-carboxylcytosine.
  JOURNAL   Science. 2011 Sep 2;333(6047):1300-3. doi: 10.1126/science.1210597. 
            Epub 2011 Jul 21.
  PUBMED    21778364
REFERENCE   2  (bases 1 to 8102)
  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"; doi: 
            "10.1126/science.1210597"; journalName: "Science"; date: "2011-09-2-
            2"; volume: "333"; issue: "6047"; pages: "1300-3"
COMMENT     Sequence Label: pcDNA3-Flag-Tet1 CD
FEATURES             Location/Qualifiers
     source          1..8102
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        84..463
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        464..667
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     618..638
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     712..731
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        712..730
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     intron          767..1339
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     CDS             1385..1408
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(3504..3522)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3505..3522)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3542..3559)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3548..3772
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3818..4246
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3905..3924)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4115..4136
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4255..4275)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4260..4589
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4440..4575
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4502..4521
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4656..5450
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4710..4729)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5320..5339
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5624..5745
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5661..5680)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5715..5734
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5794..5810)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5794..5810)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5807..5829)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5818..5834
                     /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)."
     promoter        complement(join(5842..5848,5849..5866,5867..5872))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5887..5908
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6025..6042)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6196..6784)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6276..6295)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6955..7746,7747..7815))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7578..7597
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7816..7920)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7995..8014)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
       61 tgtacgggcc agatatacgc gttgacattg attattgact agttattaat agtaatcaat
      121 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
      181 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
      241 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      301 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      361 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      421 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
      481 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      541 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      601 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      661 cagagctctc tggctaacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac
      721 tcactatagg gagacccaag ctagagtcga tcctgagaac ttcagggtga gtttggggac
      781 ccttgattgt tctttctttt tcgctattgt aaaattcatg ttatatggag ggggcaaagt
      841 tttcagggtg ttgtttagaa tgggaagatg tcccttgtat caccatggac cctcatgata
      901 attttgtttc tttcactttc tactctgttg acaaccattg tctcctctta ttttcttttc
      961 attttctgta actttttcgt taaactttag cttgcatttg taacgaattt ttaaattcac
     1021 ttttgtttat ttgtcagatt gtaagtactt tctctaatca cttttttttc aaggcaatca
     1081 gggtatatta tattgtactt cagcacagtt ttagagaaca attgttataa ttaaatgata
     1141 aggtagaata tttctgcata taaattctgg ctggcgtgga aatattctta ttggtagaaa
     1201 caactacatc ctggtcatca tcctgccttt ctctttatgg ttacaatgat atacactgtt
     1261 tgagatgagg ataaaatact ctgagtccaa accgggcccc tctgctaacc atgttcatgc
     1321 cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt tgtgctgtcg accccaagct
     1381 tatggactac aaagacgatg acgacaaggg atccgaagct gcaccctgtg actgtgatgg
     1441 aggtacacaa aaagaaaaag gcccatatta tacacacctt ggggcaggac caagtgtggc
     1501 tgctgtcagg gagctcatgg agactaggtt tggccagaag gggaaggcaa tccggattga
     1561 gaagatagtg ttcacgggga aggaagggaa gagctctcag ggctgcccgg tcgccaagtg
     1621 ggtgatcaga agaagtggtc ctgaagagaa gcttatttgt ttggttcgtg agcgtgtaga
     1681 ccatcactgt tcgacggctg tgatagttgt ccttatcctg ctgtgggaag gtatccctcg
     1741 cctgatggct gaccgcctgt acaaagagct cactgagaac ttgaggtcct acagcggaca
     1801 tcccacagac cgaagatgta ccctcaacaa aaagcgtacc tgcacctgtc aaggcatcga
     1861 cccaaaaacc tgcggagcgt ccttctcctt tggctgttcg tggagcatgt atttcaacgg
     1921 ctgtaagttt gggaggagtg aaaaccccag aaaattcaga cttgctccaa actacccctt
     1981 acataactac tataagagaa ttactggaat gagttctgaa ggaagtgacg tgaaaaccgg
     2041 gtggatcatt ccagaccgca agaccctcat aagcagagag gaaaaacagc ttgaaaagaa
     2101 tttacaagaa ttggctacag tattagctcc actttacaag cagatggctc cagttgctta
     2161 tcaaaatcag gtggaatatg aagaagttgc tggagactgt cgacttggaa atgaagaggg
     2221 gcgtcctttc tctggtgtca cctgttgcat ggatttttgt gcccattctc acaaggacat
     2281 tcacaacatg cacaacggaa gcaccgtggt gtgtacgttg attcgagcag atggccgtga
     2341 cacaaattgt cccgaggatg aacaactcca cgtcctgcca ctataccggc ttgcagacac
     2401 tgatgaattt ggctccgtgg aagggatgaa ggccaaaatc aaatctgggg ccatccaagt
     2461 caatgggcca accaggaaga ggcgactacg ttttactgag cctgttcctc gatgtgggaa
     2521 gagggccaaa atgaagcaga accacaataa atcaggttca cacaacacta agagcttttc
     2581 atcagcctca tctacttctc acctagtgaa agacgaatct acagacttct gtcccctgca
     2641 ggcttcctcc gcagaaacat ctacctgtac gtacagtaaa acagcctcag gtgggtttgc
     2701 agaaacaagt agtattctcc actgcacaat gccttctgga gcacacagtg gtgctaatgc
     2761 agctgctggg gaatgtactg gaacggtgca gcctgccgag gtggctgctc atcctcacca
     2821 gtctcttccc acagccgatt ctcccgttca tgctgagcct ctcactagtc catctgagca
     2881 gctaacttct aaccagtcaa accagcagct ccctctcctc agcaattctc agaaactggc
     2941 ttcctgtcag gtggaagatg agcggcaccc tgaagcggat gagcctcagc accccgagga
     3001 cgataacttg cctcaacttg atgaattctg gtcagacagt gaggagatct acgccgatcc
     3061 ttcctttggt ggcgtggcga tagcacccat tcacggctcg gtgctcattg agtgcgctcg
     3121 gaaggagctt catgctacca cctctttgcg ctcccccaaa cgaggggtcc cttttcgtgt
     3181 gtcccttgta ttctaccagc acaaaagcct aaacaagcct aatcatggtt ttgatatcaa
     3241 caaaattaag tgtaaatgca aaaaagtaac gaaaaaaaag cccgcagacc gggagtgtcc
     3301 tgatgtatcc cccgaagcca atttatcaca ccaaattcct tctcgagttg catcaacctt
     3361 aacccgagac aatgttgtta ccgtgtcccc atactctctc actcatgttg cgggacccta
     3421 caatcgttgg gtctaaaaag ggcgaattct gcagatatcc atcacactgg cggccgctcg
     3481 agcatgcatc tagagggccc tattctatag tgtcacctaa atgctagagc tcgctgatca
     3541 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     3601 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     3661 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     3721 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     3781 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     3841 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     3901 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3961 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4021 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4081 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4141 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     4201 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     4261 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4321 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4381 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4441 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4501 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     4561 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     4621 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     4681 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     4741 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     4801 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     4861 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     4921 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     4981 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     5041 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     5101 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     5161 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     5221 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     5281 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     5341 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     5401 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     5461 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     5521 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     5581 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     5641 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     5701 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     5761 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     5821 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     5881 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     5941 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     6001 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     6061 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     6121 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     6181 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     6241 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6301 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6361 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     6421 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6481 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6541 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6601 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     6661 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     6721 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6781 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6841 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6901 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6961 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     7021 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     7081 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     7141 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     7201 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     7261 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     7321 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     7381 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     7441 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     7501 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7561 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7621 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     7681 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7741 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7801 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7861 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7921 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     7981 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     8041 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     8101 gg
//
