LOCUS Exported 7052 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION For expression of METTL3 (methyltransferase-like 3) in human cells. ACCESSION . VERSION . KEYWORDS pcDNA3/Flag-METTL3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7052) AUTHORS Liu J, Yue Y, Han D, Wang X, Fu Y, Zhang L, Jia G, Yu M, Lu Z, Deng X, Dai Q, Chen W, He C TITLE A METTL3-METTL14 complex mediates mammalian nuclear RNA N6-adenosine methylation. JOURNAL Nat Chem Biol. 2014 Feb;10(2):93-5. doi: 10.1038/nchembio.1432. Epub 2013 Dec 6. PUBMED 24316715 REFERENCE 2 (bases 1 to 7052) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7052 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(32..51) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 223..602 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 603..806 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 757..777 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 851..870 /label=T7 /note="T7 promoter, forward primer" promoter 851..869 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 886..909 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" promoter complement(2665..2683) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(2666..2683) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(2703..2720) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2709..2933 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2979..3407 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3066..3085) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3276..3297 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3416..3436) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3460..3670 /label=SV40 promoter /note="SV40 early promoter" promoter 3482..3678 /label=SV40 promoter /note="SV40 early promoter" rep_origin 3529..3664 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3591..3610 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3745..4539 /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(3799..3818) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4409..4428 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4713..4834 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4750..4769) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4804..4823 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4883..4899) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4883..4899) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4896..4918) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4907..4923 /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(4931..4961) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4976..4997 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5114..5131) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5285..5873) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5365..5384) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6044..6904) /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 6667..6686 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6905..7009) /gene="bla" /label=AmpR promoter ORIGIN 1 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa 61 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt 121 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc 181 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta 241 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg 301 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga 361 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat 421 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa 481 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca 541 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca 601 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat 661 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg 721 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac 781 ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact gcttactggc 841 ttatcgaaat taatacgact cactataggg agacccaagc ttatggatta caaggatgac 901 gatgacaagt cggacacgtg gagctctatc caggcccaca agaagcagct ggactctctg 961 cgggagaggc tgcagcggag gcggaagcag gactcggggc acttggatct acggaatcca 1021 gaggcagcat tgtctccaac cttccgtagt gacagcccag tgcctactgc acccacctct 1081 ggtggcccta agcccagcac agcttcagca gttcctgaat tagctacaga tcctgagtta 1141 gagaagaagt tgctacacca cctctctgat ctggccttaa cattgcccac tgatgctgtg 1201 tccatctgtc ttgccatctc cacgccagat gctcctgcca ctcaagatgg ggtagaaagc 1261 ctcctgcaga agtttgcagc tcaggagttg attgaggtaa agcgaggtct cctacaagat 1321 gatgcacatc ctactcttgt aacctatgct gaccattcca agctctctgc catgatgggt 1381 gctgtggcag aaaagaaggg ccctggggag gtagcaggga ctgtcacagg gcagaagcgg 1441 cgtgcagaac aggactcgac tacagtagct gcctttgcca gttcgttagt ctctggtctg 1501 aactcttcag catcggaacc agcaaaggag ccagccaaga aatcaaggaa acatgctgcc 1561 tcagatgttg atctggagat agagagcctt ctgaaccaac agtccactaa ggaacaacag 1621 agcaagaagg tcagtcagga gatcctagag ctattaaata ctacaacagc caaggaacaa 1681 tccattgttg aaaaatttcg ctctcgaggt cgggcccaag tgcaagaatt ctgtgactat 1741 ggaaccaagg aggagtgcat gaaagccagt gatgctgatc gaccctgtcg caagctgcac 1801 ttcagacgaa ttatcaataa acacactgat gagtctttag gtgactgctc tttccttaat 1861 acatgtttcc acatggatac ctgcaagtat gttcactatg aaattgatgc ttgcatggat 1921 tctgaggccc ctggcagcaa agaccacacg ccaagccagg agcttgctct tacacagagt 1981 gtcggaggtg attccagtgc agaccgactc ttcccacctc agtggatctg ttgtgatatc 2041 cgctacctgg acgtcagtat cttgggcaag tttgcagttg tgatggctga cccaccctgg 2101 gatattcaca tggaactgcc ctatgggacc ctgacagatg atgagatgcg caggctcaac 2161 atacccgtac tacaggatga tggctttctc ttcctctggg tcacaggcag ggccatggag 2221 ttggggagag aatgtctaaa tctctggggg tatgaacggg tagatgaaat tatttgggtg 2281 aagacaaatc aactgcaacg catcattcgg acaggccgta caggtcactg gttgaaccat 2341 gggaaggaac actgcttggt tggtgtcaaa ggaaatcccc aaggcttcaa ccagggtctg 2401 gattgtgatg tgatcgtagc tgaggttcgt tccaccagtc ataaaccaga tgaaatctat 2461 ggcatgattg aaagactatc tcctggcact cgcaagattg agttatttgg acgaccacac 2521 aatgtgcaac ccaactggat cacccttgga aaccaactgg atgggatcca cctactagac 2581 ccagatgtgg ttgcacggtt caagcaaagg tacccagatg gtatcatctc taaacctaag 2641 aatttatagt ctagagggcc ctattctata gtgtcaccta aatgctagag ctcgctgatc 2701 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc 2761 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc 2821 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg 2881 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga 2941 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt 3001 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc 3061 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca 3121 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc 3181 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt 3241 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac 3301 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc 3361 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat 3421 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc 3481 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta 3541 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca 3601 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga 3661 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa 3721 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg 3781 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct 3841 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac 3901 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg 3961 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg 4021 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa 4081 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca 4141 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt 4201 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc 4261 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc 4321 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg 4381 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt 4441 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag 4501 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa 4561 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct 4621 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg 4681 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt 4741 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta 4801 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta 4861 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca 4921 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag 4981 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt 5041 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 5101 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 5161 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 5221 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 5281 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 5341 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 5401 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 5461 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 5521 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 5581 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 5641 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 5701 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag 5761 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 5821 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 5881 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 5941 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt 6001 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca 6061 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg 6121 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac 6181 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg 6241 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc 6301 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta 6361 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac 6421 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc 6481 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac 6541 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact 6601 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa 6661 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt 6721 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca 6781 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa 6841 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac 6901 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg 6961 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc 7021 gaaaagtgcc acctgacgtc gacggatcgg ga //