LOCUS       sequence-177685-        7579 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse TET2 N500 truncation in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7579)
  AUTHORS   Nakagawa T, Lv L, Nakagawa M, Yu Y, Yu C, D'Alessio AC, Nakayama K, 
            Fan HY, Chen X, Xiong Y
  TITLE     CRL4(VprBP) E3 Ligase Promotes Monoubiquitylation and Chromatin 
            Binding of TET Dioxygenases.
  JOURNAL   Mol Cell. 2015 Jan 22;57(2):247-60. doi: 
            10.1016/j.molcel.2014.12.002. Epub 2014 Dec 31.
  PUBMED    25557551
REFERENCE   2  (bases 1 to 7579)
  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: 
            "10.1016/j.molcel.2014.12.002"; journalName: "Mol Cell"; date: 
            "2015-01-22- 22"; volume: "57"; issue: "2"; pages: "247-60"
COMMENT     Sequence Label: pcDNA3-FLAG-mTET2 (N500)
FEATURES             Location/Qualifiers
     source          1..7579
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     4..25
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(144..164)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        149..478
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      329..464
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     391..410
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             545..1339
                     /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(599..618)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1209..1228
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1513..1634
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1550..1569)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1604..1623
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1683..1699)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1683..1699)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1696..1718)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1707..1723
                     /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(1731..1737,1738..1755,1756..1761))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1776..1797
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1914..1931)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2085..2673)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2165..2184)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2844..3635,3636..3704))
                     /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     3467..3486
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3705..3809)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3884..3903)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4075..4454
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4455..4658
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4609..4629
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4703..4722
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4703..4721
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     intron          4758..5330
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     CDS             5376..5399
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(6972..6990)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6973..6990)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(7010..7027)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7016..7240
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(7286..7579,1..135)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7373..7392)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
       61 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
      121 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
      181 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
      241 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
      301 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
      361 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
      421 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
      481 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
      541 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
      601 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
      661 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
      721 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
      781 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
      841 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
      901 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
      961 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     1021 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     1081 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     1141 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     1201 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     1261 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     1321 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     1381 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     1441 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     1501 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     1561 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     1621 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     1681 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     1741 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     1801 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     1861 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     1921 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     1981 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     2041 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2101 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2161 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2221 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2281 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2341 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2401 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2461 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2521 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2581 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2641 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2701 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     2761 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     2821 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     2881 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     2941 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     3001 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     3061 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     3121 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     3181 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3241 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     3301 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     3361 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     3421 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     3481 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     3541 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     3601 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     3661 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     3721 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     3781 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     3841 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
     3901 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
     3961 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
     4021 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
     4081 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     4141 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     4201 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     4261 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     4321 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     4381 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     4441 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     4501 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     4561 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     4621 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
     4681 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctagagtcg
     4741 atcctgagaa cttcagggtg agtttgggga cccttgattg ttctttcttt ttcgctattg
     4801 taaaattcat gttatatgga gggggcaaag ttttcagggt gttgtttaga atgggaagat
     4861 gtcccttgta tcaccatgga ccctcatgat aattttgttt ctttcacttt ctactctgtt
     4921 gacaaccatt gtctcctctt attttctttt cattttctgt aactttttcg ttaaacttta
     4981 gcttgcattt gtaacgaatt tttaaattca cttttgttta tttgtcagat tgtaagtact
     5041 ttctctaatc actttttttt caaggcaatc agggtatatt atattgtact tcagcacagt
     5101 tttagagaac aattgttata attaaatgat aaggtagaat atttctgcat ataaattctg
     5161 gctggcgtgg aaatattctt attggtagaa acaactacat cctggtcatc atcctgcctt
     5221 tctctttatg gttacaatga tatacactgt ttgagatgag gataaaatac tctgagtcca
     5281 aaccgggccc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     5341 acgtgctggt ttgtgctgtc gaccccaagc ttatggacta caaagacgat gacgacaagg
     5401 gatccactag taacggccgc cagtgtgctg gaattcgccc tttagaacag gacagaacca
     5461 cccatgctga gggcaccaga ctgagtccat tcctgatagc accaccttct cccatcagcc
     5521 atacagaacc tctggctgtc aaactccaga atggaagccc gttagcagag agacctcatc
     5581 cagaagtaaa tggagacacc aagtggcaat cttcccaaag ctgttatgga atatcccaca
     5641 tgaaaggcag ccagagcagt catgagagtc cacatgaaga cagagggtat tccaggtgct
     5701 tacagaatgg agggataaaa cgcacagtca gtgaaccatc tctctctggg ctccatccaa
     5761 acaagatatt gaaactagac caaaaagcta agggagaaag caatatcttc gaggaaagcc
     5821 aagaaagaaa ccacggtaaa agcagccgtc agccaaatgt ctccggacta agtgataatg
     5881 gagagcccgt gacctcgacc acccaggaaa gttcaggtgc agatgctttc ccaacacgga
     5941 actacaatgg agttgaaatt caggttctca acgagcagga aggggaaaaa ggcaggagcg
     6001 ttacattact taaaaacaag attgtgctaa tgcctaatgg tgctacagtt tctgcccatt
     6061 ctgaggagaa cacacgtggt gaactcctgg aaaaaacaca gtgttatcca gattgtgtct
     6121 ccattgccgt gcagagtacc gcatctcatg taaacacccc tagcagtcag gctgctatcg
     6181 agttgtccca cgagatccct caaccatcgc ttacctcagc gcagatcaat ttctcccaga
     6241 cctcaagctt gcagctgcct ccagagccag ctgcaatggt gactaaggcc tgtgatgctg
     6301 ataatgccag taaaccagct atagtaccag gtacctgtcc ttttcagaaa gcagaacacc
     6361 aacaaaagtc agctttggac ataggcccat ctcgtgcaga aaacaaaacc atccaaggaa
     6421 gcatggagct atttgctgaa gaatactatc ctagttccga ccggaatttg caagcttcgc
     6481 atggcagctc tgaacagtat tcaaagcaaa aggaaaccaa tggtgcttac ttcaggcaaa
     6541 gctcgaagtt ccctaaagat tccatctctc ccactactgt gaccccaccg tcacaatcac
     6601 ttcttgctcc ccgtcttgtt cttcagcctc ctttagaagg aaaaggcgct ctaaatgatg
     6661 tagctttgga agaacaccat gactacccca accgaagcaa ccgaactctt ttaagggaag
     6721 ggaaaataga ccatcaaccc aagacatcat ctagccagag tctgaatcca tctgtacata
     6781 cacccaaccc ccccttgatg cttccagaac agcatcagaa tgattgtggc tcaccgagcc
     6841 ctgaaaagtc aagaaaaatg tcagaatatc tcatgtatta cctgccaaat catggccaca
     6901 gtggaggttt acaagaacat agccaatacc tgatggggca ctgactcgag catgcatcta
     6961 gagggcccta ttctatagtg tcacctaaat gctagagctc gctgatcagc ctcgactgtg
     7021 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     7081 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     7141 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     7201 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     7261 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     7321 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     7381 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     7441 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     7501 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     7561 ttggagtcca cgttcttta
//
