LOCUS Exported 8755 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses mouse TET2 CD mutant in mammalian cells. ACCESSION . VERSION . KEYWORDS pcDNA3-FLAG-mTET2 CD mutant SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8755) 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 8755) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8755 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 213..233 /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin complement(242..670) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(352..373) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 564..583 /label=F1ori-R /note="F1 origin, reverse primer" polyA_signal 716..940 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind 929..946 /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" promoter 966..984 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 966..983 /label=SP6 /note="SP6 promoter, forward primer" CDS complement(3733..3756) /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" intron 3802..4374 /label=-beta-globin intron /note="intron from rabbit beta-globin gene" primer_bind complement(4410..4429) /label=T7 /note="T7 promoter, forward primer" promoter complement(4411..4429) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" promoter complement(4474..4677) /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind complement(4503..4523) /label=CMV-F /note="Human CMV immediate early promoter, forward primer" enhancer 4678..5057 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" primer_bind 5229..5248 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" promoter 5323..5427 /gene="bla" /label=AmpR promoter CDS 5428..6288 /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 complement(5646..5665) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6459..7047 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6948..6967 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 7201..7218 /label=L4440 /note="L4440 vector, forward primer" protein_bind 7335..7356 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 7371..7401 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7409..7425 /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)." primer_bind 7414..7436 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 7433..7449 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 7433..7449 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" polyA_signal 7498..7619 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(7509..7528) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 7563..7582 /label=SV40pA-R /note="SV40 polyA, reverse primer" CDS complement(7793..8587) /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(7904..7923) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 8514..8533 /label=Neo-R /note="Neomycin resistance gene, reverse primer" promoter complement(8654..228) /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 8668..48 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(8722..8741) /label=SV40pro-F /note="SV40 promoter/origin, forward primer" ORIGIN 1 gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag 61 gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg 121 ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc tttgcatact 181 tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat tccacagaat 241 taattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc 301 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg 361 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat 421 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc 481 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag 541 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg 601 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta 661 cagggcgcgt ggggataccc cctagagccc cagctggttc tttccgcctc agaagccata 721 gagcccaccg catccccagc atgcctgcta ttgtcttccc aatcctcccc cttgctgtcc 781 tgccccaccc caccccccag aatagaatga cacctactca gacaatgcga tgcaatttcc 841 tcattttatt aggaaaggac agtgggagtg gcaccttcca gggtcaagga aggcacgggg 901 gaggggcaaa caacagatgg ctggcaacta gaaggcacag tcgaggctga tcagcgagct 961 ctagcattta ggtgacacta tagaataggg ccctctagat gcatgctcga gcggccgcca 1021 gtgtgatgga tatctgcaga attcgccctt tcatacaaat gtgttgtaag gccctgtgac 1081 ctgagtgaaa gcatatggtg atgtagtcac ggtagaatcc gtagtcacag accctgtgtt 1141 ctcagcaaga gactggatga aacgcaggta actgggttcc tgtggccctg tgggctcacg 1201 cttttcctgt ttgccatgat ttttctgaga cacgtggtct gatccattct ttccgcactc 1261 ttcctctttc cgggcctttt cagccatttt ggcttcccag agagccaaac aatgttttgg 1321 tagaaacaaa ttcttatgcc tatacagtac aagtgagatc ctggtggggt gattccggtc 1381 gggatcgttt actttggttg tggcatgaac ctcacacttt gcacactcaa taagaattga 1441 cccatgagtt ggggctatag ccacccctcc aatgcaagga tcctgaaagt tgtgctcact 1501 atctgaccaa tactcaattt cctgcacagc agctgcatcc tggcctgaca cctcaggctg 1561 cttctcctga ctgctgccag tcagactgta taatagttct tgggcagaag cagttctatc 1621 atgattaaac cctggaagca ctcttgagag cccattggct atgttgtcat tctccttgtt 1681 gtggaaggcg tggctggaac tgctccccga agctgctgcc gtgtagctgt agaccgtgtg 1741 agagggtaga tgatgctcac tggttttgta gtcagtgtgt gggtggctgt atggtgatct 1801 ggaggcagct cccatgaaat gactatccat cttgggggtg gggtaaggag aaaacgttgc 1861 taggtggtgt acatttggtt ttagggtgga gttagtagtg aaggcatctc tctgcatatt 1921 ttggtttcca taacttaagt acttagaggg actgtcccca aacgtctgtt ggtgaagggt 1981 gtggatgggt ggtaagttct gattggtgag atggtcctgg tttggatatc tatgtagatc 2041 cctggactga gcctgggggg aataagaacc taagaaaggg gaaccattgt ccactggcac 2101 actcccattg tatggaaaag gtgcatattg gttattctga ttcaaaagcc caaggtaggg 2161 gttcatgtaa ttagaaggat tcaaatatga acccgaggca tgagatgaag tggggtaaaa 2221 gttcacatgg ttggtatctc cataaatatc tgaggtgtgt gctgagctgg gataaggact 2281 gtgaggagta ggctgtctcg tgtagacact ggtggatcca gaacaatgag aaccaacaga 2341 ctgagagtta ccgggcatga tatgctgtgg ctgtggctgt ggctgaggtg tcgtctgcgg 2401 ctgcggctga ggtggctgcg gctgctgtgg cctctgccct tgctgaaggt gcctctggag 2461 tgttggtggc tgccggatga ccgggcccga aagctgcggt tgtgctgtca tttgtttcat 2521 atggcttgca ttttccatca gctttgtatg tgaggaagac tttcccttct cagtcctact 2581 ggagcagttc tccagagagg aatgctttcg agctgctttc ttggtcttgg ctttcttggg 2641 ctccgctttc ttcttgcaac tcttgggcag ctctcctatc cttatgaccc ttctcctccg 2701 aaatgactgc agaacctcaa tggaccccat ccgtatcttc ttctcctggc cttccgtact 2761 cccaaactca tcctcagggg cgatgatgta cataggcagc acgtggaact gctcatcctc 2821 aggcttagct ccgacttctc gattgtcttc tctattgagg gtgaccacca ctgtactgcc 2881 atttggcatg ttctgctggg ctctgtagga atgagcagag aagtccaaac atgcagtgac 2941 tcctgagaat ggccggcctt ccttcagacc caaacagcag tctggggctt ggtgttcaaa 3001 ttcaacctga ttattgtatg catcgggtgc aagcttcttg tatattggag caatgacagt 3061 agccaggttt tgcaaatgag aacctagtct ctcttcctct tttggctcag ctccatgtag 3121 cctaaatttc cttggtttct tgcttctggc aaacttacat ccattatagt acatgctcca 3181 agaacaacca aaagaaaagg aggcaccaca ggtctctgga ttctcaccct gacaacaaca 3241 gtttctgggt ggtgattgct ttttcttcgt ttcattctga gaacagcgac ggttggtgca 3301 tatgccacac ttgcccaaga tatctgtaag ttctgagtag agttctgatg cgagtagctt 3361 tgggattccg tcccacaaca tgatggcaat taccatcacc gccgtctcac atgtgtggtt 3421 aggtcgcact cgtaccaaac acagtagttt ctcctcctca ctcgatctcc gatataccca 3481 tttagcaata ggacatccct gagagctctt gccttcttta ccagtatata tgactttttc 3541 aatcctaata gctttaccct tctctccata cctttcttcc atgagtgttc taatagctgc 3601 cacatcagga ccagctccca gatgggtata ataaggagct tcatctttgt ctggattgca 3661 tccttcacat ttgccattct gactttgtaa agggcgaatt ccagcacact ggcggccgtt 3721 actagtggat cccttgtcgt catcgtcttt gtagtccata agcttggggt cgacagcaca 3781 aaccagcacg ttgcccagga gctgtaggaa aaagaagaag gcatgaacat ggttagcaga 3841 ggggcccggt ttggactcag agtattttat cctcatctca aacagtgtat atcattgtaa 3901 ccataaagag aaaggcagga tgatgaccag gatgtagttg tttctaccaa taagaatatt 3961 tccacgccag ccagaattta tatgcagaaa tattctacct tatcatttaa ttataacaat 4021 tgttctctaa aactgtgctg aagtacaata taatataccc tgattgcctt gaaaaaaaag 4081 tgattagaga aagtacttac aatctgacaa ataaacaaaa gtgaatttaa aaattcgtta 4141 caaatgcaag ctaaagttta acgaaaaagt tacagaaaat gaaaagaaaa taagaggaga 4201 caatggttgt caacagagta gaaagtgaaa gaaacaaaat tatcatgagg gtccatggtg 4261 atacaaggga catcttccca ttctaaacaa caccctgaaa actttgcccc ctccatataa 4321 catgaatttt acaatagcga aaaagaaaga acaatcaagg gtccccaaac tcaccctgaa 4381 gttctcagga tcgactctag cttgggtctc cctatagtga gtcgtattaa tttcgataag 4441 ccagtaagca gtgggttctc tagttagcca gagagctctg cttatataga cctcccaccg 4501 tacacgccta ccgcccattt gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc 4561 ccgttgattt tggtgccaaa acaaactccc attgacgtca atggggtgga gacttggaaa 4621 tccccgtgag tcaaaccgct atccacgccc attgatgtac tgccaaaacc gcatcaccat 4681 ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc ataaggtcat 4741 gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag ggggcgtact 4801 tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat actccaccca 4861 ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca ttattgacgt 4921 caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac 4981 gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact attaataact 5041 agtcaataat caatgtcaac gcgtatatct ggcccgtaca tcgcgaagca gcgcaaaacg 5101 cctaacccta agcagattct tcatgcaatt gtcggtcaag ccttgccttg ttgtagctta 5161 aattttgctc gcgcactact cagcgacctc caacacacaa gcagggagca gatactggct 5221 taactatgcg gcatcagagc agattgtact gagagtgcac cataggggat cgggagatct 5281 cccgatccgt cgacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt 5341 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg 5401 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt 5461 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta 5521 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc 5581 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa 5641 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc 5701 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt 5761 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact 5821 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac 5881 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata 5941 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta 6001 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg 6061 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat 6121 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt 6181 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga 6241 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa 6301 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag 6361 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac 6421 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc 6481 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat 6541 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat 6601 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct 6661 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt 6721 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg 6781 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta 6841 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg 6901 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg 6961 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc 7021 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg 7081 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat 7141 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc 7201 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg 7261 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt 7321 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt 7381 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac 7441 agctatgacc atgattacgc caagctctag ctagaggtcg acggtataca gacatgataa 7501 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt 7561 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagttg 7621 gggtgggcga agaactccag catgagatcc ccgcgctgga ggatcatcca gccggcgtcc 7681 cggaaaacga ttccgaagcc caacctttca tagaaggcgg cggtggaatc gaaatctcgt 7741 gatggcaggt tgggcgtcgc ttggtcggtc atttcgaacc ccagagtccc gctcagaaga 7801 actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa 7861 gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca 7921 acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa 7981 agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc acgacgagat 8041 cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag ttcggctggc gcgagcccct 8101 gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc 8161 gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca 8221 gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca 8281 ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa 8341 cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct 8401 cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc 8461 cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt 8521 catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt 8581 caatcatgcg aaacgatcct catcctgtct cttgatcaga tccgaaaatg gatatacaag 8641 ctcccgggag ctttttgcaa aagcctaggc ctccaaaaaa gcctcctcac tacttctgga 8701 atagctcaga ggcagaggcg gcctcggcct ctgcataaat aaaaaaaatt agtca //