LOCUS       sequence-217039-        6572 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-terminally FLAG-tagged human RNMT 121-476 in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6572)
  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 6572)
  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-FLAG-RNMT 121-476
FEATURES             Location/Qualifiers
     source          1..6572
                     /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..741
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        complement(1917..1935)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1918..1935)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1955..1972)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1961..2185
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2231..2659
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2318..2337)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2528..2549
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2668..2688)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2673..3002
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2853..2988
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2915..2934
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3069..3863
                     /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(3123..3142)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3733..3752
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4037..4158
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4074..4093)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4128..4147
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4207..4223)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4207..4223)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4220..4242)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4231..4247
                     /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(4255..4261,4262..4279,4280..4285))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4300..4321
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4438..4455)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4609..5197)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4689..4708)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5368..6159,6160..6228))
                     /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     5991..6010
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6229..6333)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6408..6427)
                     /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 agcggccata tcgaaggtcg tcatgatggc actcaaaata agagaaaaat agcacttgag
      781 gatgttcctg aaaagcagaa aaatctggaa gaaggacaca gctcaacagt ggctgcccat
      841 tacaatgaac ttcaggaagt tggtttggag aagcgtagtc aaagtcgtat tttttaccta
      901 agaaacttta ataattggat gaaaagtgtt ctcattggag aatttttgga aaaggtacga
      961 cagaagaaaa aacgtgatat cactgttttg gacctgggat gtggtaaagg tggagatttg
     1021 ctgaaatgga aaaaaggaag aattaacaag ctagtttgta ctgatattgc cgatgtttct
     1081 gtcaaacagt gtcagcagcg gtatgaggac atgaaaaatc gtcgtgatag tgaatatatt
     1141 ttcagtgcag aatttataac tgctgacagc tcaaaggaac ttctgattga caaatttcgt
     1201 gacccacaaa tgtgttttga catctgcagt tgtcagtttg tctgtcatta ctcatttgag
     1261 tcttatgagc aggctgacat gatgctgaga aatgcgtgtg agagacttag ccctgggggc
     1321 tattttattg gtactactcc caatagcttt gaattgataa gacgccttga agcttcagaa
     1381 acagaatcat ttggaaatga aatatatact gtgaaatttc agaagaaagg agattatcct
     1441 ttatttggct gcaaatatga cttcaacttg gaaggtgttg tggatgttcc tgaattcttg
     1501 gtctattttc cattgctaaa tgaaatggca aagaagtaca atatgaaact agtctacaaa
     1561 aaaacatttc tggaattcta cgaagaaaag attaagaaca atgaaaataa aatgctctta
     1621 aaacgaatgc aggccttgga gccatatcct gcaaatgaga gttctaaact tgtctctgag
     1681 aaggtggatg actatgaaca tgcagcaaag tacatgaaga acagtcaagt aaggttacct
     1741 ttgggaacct taagtaaatc agaatgggaa gctacaagta tttacttggt gtttgccttt
     1801 gagaaacagc agtgatgacg agctcggatc cactagtaac ggccgccagt gtgctggaat
     1861 tctgcagata tccatcacac tggcggccgc tcgagcatgc atctagaggg ccctattcta
     1921 tagtgtcacc taaatgctag agctcgctga tcagcctcga ctgtgccttc tagttgccag
     1981 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     2041 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     2101 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     2161 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     2221 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2281 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     2341 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2401 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     2461 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2521 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     2581 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     2641 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     2701 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     2761 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     2821 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     2881 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     2941 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3001 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     3061 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     3121 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     3181 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     3241 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     3301 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     3361 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     3421 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     3481 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     3541 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     3601 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     3661 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     3721 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     3781 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     3841 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     3901 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     3961 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     4021 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     4081 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     4141 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     4201 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     4261 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     4321 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     4381 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     4441 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     4501 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     4561 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4621 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4681 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4741 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4801 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4861 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4921 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4981 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5041 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5101 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     5161 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     5221 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     5281 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     5341 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     5401 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     5461 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     5521 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     5581 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     5641 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     5701 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     5761 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     5821 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     5881 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     5941 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     6001 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     6061 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     6121 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     6181 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     6241 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     6301 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     6361 cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct
     6421 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     6481 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
     6541 ctgcttaggg ttaggcgttt tgcgctgctt cg
//
