LOCUS       sequence-326689-        7016 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of PRMT3 with an N-terminal FLAG tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7016)
  AUTHORS   Kaniskan HU, Szewczyk MM, Yu Z, Eram MS, Yang X, Schmidt K, Luo X, 
            Dai M, He F, Zang I, Lin Y, Kennedy S, Li F, Dobrovetsky E, Dong A, 
            Smil D, Min SJ, Landon M, Lin-Jones J, Huang XP, Roth BL, Schapira 
            M, Atadja P, Barsyte-Lovejoy D, Arrowsmith CH, Brown PJ, Zhao K, Jin
            J, Vedadi M
  TITLE     A potent, selective and cell-active allosteric inhibitor of protein 
            arginine methyltransferase 3 (PRMT3).
  JOURNAL   Angew Chem Int Ed Engl. 2015 Apr 20;54(17):5166-70. doi: 
            10.1002/anie.201412154. Epub 2015 Feb 27.
  PUBMED    25728001
REFERENCE   2  (bases 1 to 7016)
  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: 
            "10.1002/anie.201412154"; journalName: "Angew Chem Int Ed Engl"; 
            date: "2015-04-20- 20"; volume: "54"; issue: "17"; pages: "5166-70"
COMMENT     Sequence Label: FLAG-PRMT3
FEATURES             Location/Qualifiers
     source          1..7016
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      679..688
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             700..714
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     promoter        complement(2335..2353)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2336..2353)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2373..2390)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2379..2603
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2649..3077
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2736..2755)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2946..2967
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3086..3106)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3091..3420
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3271..3406
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3333..3352
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3487..4281
                     /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(3541..3560)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4151..4170
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4455..4576
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4492..4511)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4546..4565
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4625..4641)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4625..4641)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4638..4660)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4649..4665
                     /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(4673..4679,4680..4697,4698..4703))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4718..4739
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4856..4873)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5027..5615)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5107..5126)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5786..6577,6578..6646))
                     /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     6409..6428
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6647..6751)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6826..6845)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgagc tcggatccgc caccatggat tacaaggatg acgacgacga taagtgctcg
      721 ttagcgtcag gcgctaccgg cggccggggc gctgtggaga atgaggagga cctgccagaa
      781 ctgtcggaca gcggggacga ggccgcctgg gaggatgagg acgatgcaga tctcccccac
      841 ggcaagcagc agaccccctg cctgttctgt aacaggttat tcacatctgc tgaagaaaca
      901 ttttcgcact gtaagtctga gcatcagttt aatattgaca gcatggttca taaacatgga
      961 cttgaatttt atggatacat taagctaata aattttatta gacttaagaa tcctacagtt
     1021 gagtacatga attccatata caacccagtg ccttgggaga aagaagagta tttgaagcca
     1081 gtattagaag atgacctttt acttcaattt gatgtagaag atctttatga accggtgtca
     1141 gtacccttct cataccccaa tggactcagt gaaaatacat ctgctgttga aaaattgaaa
     1201 catatggaag ccagggcact gtctgctgaa gccgcattgg ccagagcacg tgaggatctg
     1261 caaaaaatga aacaatttgc tcaggatttt gtgatgcaca cagatgtcag aacctgctcg
     1321 tcatctacta gtgtcattgc ggacctccag gaggatgagg atggtgttta tttcagctca
     1381 tacgggcatt atgggataca tgaagaaatg ctaaaggaca aaatacgaac agaaagctac
     1441 cgagatttca tataccaaaa tccacatatc ttcaaagaca aggtagtttt ggatgttggg
     1501 tgtggaactg gaattctctc tatgtttgct gctaaagctg gggcgaagaa ggttcttgga
     1561 gttgatcaat ctgaaatact ttaccaggca atggatatta taagactaaa taaacttgaa
     1621 gatactatta cactaattaa aggaaagatt gaagaagttc atcttcctgt agaaaaagta
     1681 gatgttatca tatctgagtg gatgggctat tttcttctgt ttgagtctat gttagattct
     1741 gtcctttatg caaagaacaa atacttggca aaaggaggct cggtctaccc tgacatttgc
     1801 actatcagcc ttgtagcagt gagtgatgtg aataaacatg ctgatagaat tgctttttgg
     1861 gatgatgtct atggcttcaa gatgtcctgc atgaagaaag cagttatccc agaagctgtt
     1921 gtggaagttt tagatccgaa gactcttatt tcagaacctt gtggtattaa gcatatagat
     1981 tgccatacga cgtctatctc agatttggaa ttttcatcag attttaccct gaaaatcaca
     2041 aggacatcca tgtgcacggc aattgctggc tactttgata tatattttga gaagaattgc
     2101 cacaacaggg tcgtgttctc tacgggccct cagagcacca aaacacactg gaaacaaaca
     2161 gtatttctac tggaaaaacc attttcagtt aaagcaggtg aagccttgaa aggaaaggtc
     2221 acagttcaca agaataagaa agatccacgt tctctcaccg tgaccctcac gttgaataat
     2281 tcaactcaaa cttatggtct ccagtgactc gagcatgcat ctagagggcc ctattctata
     2341 gtgtcaccta aatgctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc
     2401 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     2461 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     2521 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     2581 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     2641 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     2701 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     2761 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     2821 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     2881 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     2941 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     3001 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     3061 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3121 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3181 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3241 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3301 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     3361 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3421 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     3481 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     3541 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     3601 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     3661 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     3721 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     3781 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     3841 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     3901 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     3961 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     4021 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     4081 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4141 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4201 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4261 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     4321 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     4381 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     4441 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     4501 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     4561 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     4621 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     4681 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     4741 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     4801 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     4861 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     4921 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4981 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5041 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5101 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5161 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5221 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5281 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5341 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5401 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5461 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5521 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5581 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     5641 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5701 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5761 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5821 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5881 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5941 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6001 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6061 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6121 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6181 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6241 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6301 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6361 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6421 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6481 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6541 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6601 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     6661 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6721 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     6781 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
     6841 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     6901 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
     6961 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgtt
//
