LOCUS       sequence-327074-        7016 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of PRMT3(E338Q) 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(E338Q)
FEATURES             Location/Qualifiers
     source          1..7016
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        7..386
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        387..590
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     541..561
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     635..654
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        635..653
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      685..694
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             706..720
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     promoter        complement(2341..2359)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2342..2359)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2379..2396)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2385..2609
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2655..3083
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2742..2761)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2952..2973
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3092..3112)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3097..3426
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3277..3412
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3339..3358
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3493..4287
                     /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(3547..3566)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4157..4176
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4461..4582
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4498..4517)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4552..4571
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4631..4647)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4631..4647)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4644..4666)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4655..4671
                     /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(4679..4685,4686..4703,4704..4709))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4724..4745
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4862..4879)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5033..5621)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5113..5132)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5792..6583,6584..6652))
                     /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     6415..6434
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6653..6757)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6832..6851)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
       61 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      121 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      181 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      241 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      301 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      361 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      421 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      481 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      541 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa
      601 ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc
      661 aagcttggta ccgagctcgg atccgccacc atggattaca aggatgacga cgacgataag
      721 tgctcgttag cgtcaggcgc taccggcggc cggggcgctg tggagaatga ggaggacctg
      781 ccagaactgt cggacagcgg ggacgaggcc gcctgggagg atgaggacga tgcagatctc
      841 ccccacggca agcagcagac cccctgcctg ttctgtaaca ggttattcac atctgctgaa
      901 gaaacatttt cgcactgtaa gtctgagcat cagtttaata ttgacagcat ggttcataaa
      961 catggacttg aattttatgg atacattaag ctaataaatt ttattagact taagaatcct
     1021 acagttgagt acatgaattc catatacaac ccagtgcctt gggagaaaga agagtatttg
     1081 aagccagtat tagaagatga ccttttactt caatttgatg tagaagatct ttatgaaccg
     1141 gtgtcagtac ccttctcata ccccaatgga ctcagtgaaa atacatctgc tgttgaaaaa
     1201 ttgaaacata tggaagccag ggcactgtct gctgaagccg cattggccag agcacgtgag
     1261 gatctgcaaa aaatgaaaca atttgctcag gattttgtga tgcacacaga tgtcagaacc
     1321 tgctcgtcat ctactagtgt cattgcggac ctccaggagg atgaggatgg tgtttatttc
     1381 agctcatacg ggcattatgg gatacatgaa gaaatgctaa aggacaaaat acgaacagaa
     1441 agctaccgag atttcatata ccaaaatcca catatcttca aagacaaggt agttttggat
     1501 gttgggtgtg gaactggaat tctctctatg tttgctgcta aagctggggc gaagaaggtt
     1561 cttggagttg atcaatctga aatactttac caggcaatgg atattataag actaaataaa
     1621 cttgaagata ctattacact aattaaagga aagattgaag aagttcatct tcctgtagaa
     1681 aaagtagatg ttatcatatc tgagtggatg ggctattttc ttctgtttca gtctatgtta
     1741 gattctgtcc tttatgcaaa gaacaaatac ttggcaaaag gaggctcggt ctaccctgac
     1801 atttgcacta tcagccttgt agcagtgagt gatgtgaata aacatgctga tagaattgct
     1861 ttttgggatg atgtctatgg cttcaagatg tcctgcatga agaaagcagt tatcccagaa
     1921 gctgttgtgg aagttttaga tccgaagact cttatttcag aaccttgtgg tattaagcat
     1981 atagattgcc atacgacgtc tatctcagat ttggaatttt catcagattt taccctgaaa
     2041 atcacaagga catccatgtg cacggcaatt gctggctact ttgatatata ttttgagaag
     2101 aattgccaca acagggtcgt gttctctacg ggccctcaga gcaccaaaac acactggaaa
     2161 caaacagtat ttctactgga aaaaccattt tcagttaaag caggtgaagc cttgaaagga
     2221 aaggtcacag ttcacaagaa taagaaagat ccacgttctc tcaccgtgac cctcacgttg
     2281 aataattcaa ctcaaactta tggtctccag tgactcgagc atgcatctag agggccctat
     2341 tctatagtgt cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg
     2401 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     2461 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     2521 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     2581 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     2641 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     2701 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     2761 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     2821 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     2881 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     2941 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     3001 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     3061 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     3121 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3181 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3241 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3301 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3361 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     3421 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     3481 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     3541 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     3601 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     3661 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     3721 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     3781 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     3841 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     3901 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     3961 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4021 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4081 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4141 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4201 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     4261 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     4321 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     4381 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     4441 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     4501 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     4561 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     4621 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     4681 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     4741 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     4801 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     4861 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4921 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4981 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5041 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5101 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5161 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5221 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5281 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5341 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5401 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5461 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5521 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5581 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5641 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5701 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5761 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5821 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5881 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5941 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6001 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6061 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     6121 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6181 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6241 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6301 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6361 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6421 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     6481 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6541 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6601 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6661 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6721 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6781 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg
     6841 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
     6901 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
     6961 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatata
//
