LOCUS       sequence-217040-        6608 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG-NES-RNMT 121-476 D203A (""Delta-N-RNMT"") in 
            mammalian cells. Overexpression inhibits cytoplasmic cap 
            methylation..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6608)
  AUTHORS   Trotman JB, Giltmier AJ, Mukherjee C, Schoenberg DR
  TITLE     RNA guanine-7 methyltransferase catalyzes the methylation of 
            cytoplasmically recapped RNAs.
  JOURNAL   Nucleic Acids Res. 2017 Oct 13;45(18):10726-10739. doi: 
            10.1093/nar/gkx801.
  PUBMED    28981715
REFERENCE   2  (bases 1 to 6608)
  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"; journalName: 
            "Nucleic Acids Res"; date: "2017-10-13- 13"; volume: "45"; issue: 
            "18"; pages: "10726-10739"
COMMENT     Sequence Label: pcDNA3-Delta-N-RNMT
FEATURES             Location/Qualifiers
     source          1..6608
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        27..406
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        407..610
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     561..581
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     655..674
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        655..673
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             694..717
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             730..756
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             766..777
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        complement(1953..1971)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1954..1971)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1991..2008)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1997..2221
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2267..2695
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2354..2373)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2564..2585
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2704..2724)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2709..3038
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2889..3024
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2951..2970
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3105..3899
                     /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(3159..3178)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3769..3788
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4073..4194
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4110..4129)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4164..4183
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4243..4259)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4243..4259)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4256..4278)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4267..4283
                     /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(4291..4297,4298..4315,4316..4321))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4336..4357
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4474..4491)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4645..5233)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4725..4744)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5404..6195,6196..6264))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMAVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6027..6046
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6265..6369)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6444..6463)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
       61 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      121 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      181 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      241 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      301 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      361 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
      421 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      481 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      541 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      601 aagcagagct ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac
      661 gactcactat agggagaccc aagcttgatg ggcgactaca aagacgatga cgacaagagc
      721 agccttcagc taccaccgct tgagagactt actcttgatg gccatatcga aggtcgtcat
      781 gatggcactc aaaataagag aaaaatagca cttgaggatg ttcctgaaaa gcagaaaaat
      841 ctggaagaag gacacagctc aacagtggct gcccattaca atgaacttca ggaagttggt
      901 ttggagaagc gtagtcaaag tcgtattttt tacctaagaa actttaataa ttggatgaaa
      961 agtgttctca ttggagaatt tttggaaaag gtacgacaga agaaaaaacg tgatatcact
     1021 gttttggccc tgggatgtgg taaaggtgga gatttgctga aatggaaaaa aggaagaatt
     1081 aacaagctag tttgtactga tattgccgat gtttctgtca aacagtgtca gcagcggtat
     1141 gaggacatga aaaatcgtcg tgatagtgaa tatattttca gtgcagaatt tataactgct
     1201 gacagctcaa aggaacttct gattgacaaa tttcgtgacc cacaaatgtg ttttgacatc
     1261 tgcagttgtc agtttgtctg tcattactca tttgagtctt atgagcaggc tgacatgatg
     1321 ctgagaaatg cgtgtgagag acttagccct gggggctatt ttattggtac tactcccaat
     1381 agctttgaat tgataagacg ccttgaagct tcagaaacag aatcatttgg aaatgaaata
     1441 tatactgtga aatttcagaa gaaaggagat tatcctttat ttggctgcaa atatgacttc
     1501 aacttggaag gtgttgtgga tgttcctgaa ttcttggtct attttccatt gctaaatgaa
     1561 atggcaaaga agtacaatat gaaactagtc tacaaaaaaa catttctgga attctacgaa
     1621 gaaaagatta agaacaatga aaataaaatg ctcttaaaac gaatgcaggc cttggagcca
     1681 tatcctgcaa atgagagttc taaacttgtc tctgagaagg tggatgacta tgaacatgca
     1741 gcaaagtaca tgaagaacag tcaagtaagg ttacctttgg gaaccttaag taaatcagaa
     1801 tgggaagcta caagtattta cttggtgttt gcctttgaga aacagcagtg atgacgagct
     1861 cggatccact agtaacggcc gccagtgtgc tggaattctg cagatatcca tcacactggc
     1921 ggccgctcga gcatgcatct agagggccct attctatagt gtcacctaaa tgctagagct
     1981 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     2041 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     2101 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     2161 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     2221 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     2281 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2341 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2401 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     2461 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     2521 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     2581 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     2641 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     2701 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     2761 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     2821 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     2881 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     2941 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     3001 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     3061 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     3121 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     3181 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     3241 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     3301 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     3361 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     3421 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     3481 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     3541 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     3601 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     3661 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     3721 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     3781 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     3841 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     3901 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     3961 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4021 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     4081 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     4141 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     4201 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     4261 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     4321 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     4381 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcagggag
     4441 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4501 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4561 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4621 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcgcaa
     4681 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4741 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4801 gtccgccttt cttccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4861 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4921 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4981 attgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5041 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     5101 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5161 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5221 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5281 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5341 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5401 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5461 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     5521 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     5581 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     5641 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     5701 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     5761 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5821 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     5881 actcatggtt atggcagcac tgcataattc tcttactgcc atgccatccg taagatgctt
     5941 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     6001 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     6061 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     6121 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6181 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6241 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6301 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6361 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     6421 gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     6481 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     6541 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg
     6601 ctgcttcg
//
