LOCUS       pcDNA3/Flag-METTL3                7052 bp ds-DNA     circular SYN 06-AUG-2018
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 Aug 6, 2018 from SnapGene Server 1.1.58
            //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
//