LOCUS       sequence-473333-        6654 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express PRMT5-NanoLuc(R) Fusion Protein in Mammalian Cells under a 
            CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6654)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6654)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: NanoBRET Assay Vector, PRMT5-NanoLuc
FEATURES             Location/Qualifiers
     source          1..6654
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1063..1072
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2997..3509
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(3597..3615)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3652..3773
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3663..3682)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3717..3736
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4018..4375
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4018..4209
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4041..4061)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4226..4361
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4390..4437
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4469..5263
                     /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(4523..4542)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5133..5152
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5327..5375
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5614..6202)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5694..5713)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6318..6601
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcgat cgccatggcg gcgatggcgg
     1081 tcgggggtgc tggtgggagc cgcgtgtcca gcgggaggga cctgaattgc gtccccgaaa
     1141 tagctgacac actaggggct gtggccaagc aggggtttga tttcctctgc atgcctgtct
     1201 tccatccgcg tttcaagagg gagttcattc aggaacctgc taagaatcgg cccggtcccc
     1261 agacacgatc agacctactg ctgtcaggaa gggactggaa tacgctaatt gtgggaaagc
     1321 tttctccatg gattcgtcca gactcaaaag tggagaagat tcgcaggaac tccgaggcgg
     1381 ccatgttaca ggagctgaat tttggtgcat atttgggtct tccagctttc ctgctgcccc
     1441 ttaatcagga agataacacc aacctggcca gagttttgac caaccacatc cacactggcc
     1501 atcactcttc catgttctgg atgcgggtac ccttggtggc accagaggac ctgagagatg
     1561 atataattga gaatgcacca actacacaca cagaggagta cagtggggag gagaaaacgt
     1621 ggatgtggtg gcacaacttc cggactttgt gtgactatag taagaggatt gcagtggctc
     1681 ttgaaattgg ggctgacctc ccatctaatc atgtcattga tcgctggctt ggggagccca
     1741 tcaaagcagc cattctcccc actagcattt tcctgaccaa taagaaggga tttcctgttc
     1801 tttctaagat gcaccagagg ctcatcttcc ggctcctcaa gttggaggtg cagttcatca
     1861 tcacaggcac caaccaccac tcagagaagg agttctgctc ctacctccaa tacctggaat
     1921 acttaagcca gaaccgtcct ccacctaatg cctatgaact ctttgccaag ggctatgaag
     1981 actatctgca gtccccgctt cagccactga tggacaatct ggaatctcag acatatgaag
     2041 tgtttgaaaa ggaccccatc aaatactctc agtaccagca ggccatctat aaatgtctgc
     2101 tagaccgagt accagaagag gagaaggata ccaatgtcca ggtactgatg gtgctgggag
     2161 caggacgggg acccctggtg aacgcttccc tgcgggcagc caagcaggcc gaccggcgga
     2221 taaagctgta tgctgtggag aaaaacccaa atgccgtggt gacgctagag aactggcagt
     2281 ttgaagaatg gggaagccaa gtgaccgtag tctcatcaga catgagggaa tgggtggctc
     2341 cagagaaagc agacatcatt gtcagtgagc ttctgggctc atttgctgac aatgaattgt
     2401 cgcctgagtg cctggatgga gcccagcact tcctaaaaga tgatggtgtg agcatccccg
     2461 gggagtacac ttcctttctg gctcccatct cttcctccaa gctgtacaat gaggtccgag
     2521 cctgtaggga gaaggaccgt gaccctgagg cccagtttga gatgccttat gtggtacggc
     2581 tgcacaactt ccaccagctc tctgcacccc agccctgttt caccttcagc catcccaaca
     2641 gagatcctat gattgacaac aaccgctatt gcaccttgga atttcctgtg gaggtgaaca
     2701 cagtactaca tggctttgcc ggctactttg agactgtgct ttatcaggac atcactctga
     2761 gtatccgtcc agagactcac tctcctggga tgttctcatg gtttcccatc ctcttcccta
     2821 ttaagcagcc cataacggta cgtgaaggcc aaaccatctg tgtgcgtttc tggcgatgca
     2881 gcaattccaa gaaggtgtgg tatgagtggg ctgtgacagc accagtctgt tctgctattc
     2941 ataaccccac aggccgctca tataccattg gcctcgtttc tctcggctcg agcggcgtct
     3001 tcacactcga agatttcgtt ggggactggc gacagacagc cggctacaac ctggaccaag
     3061 tccttgaaca gggaggtgtg tccagtttgt ttcagaatct cggggtgtcc gtaactccga
     3121 tccaaaggat tgtcctgagc ggtgaaaatg ggctgaagat cgacatccat gtcatcatcc
     3181 cgtatgaagg tctgagcggc gaccaaatgg gccagatcga aaaaattttt aaggtggtgt
     3241 accctgtgga tgatcatcac tttaaggtga tcctgcacta tggcacactg gtaatcgacg
     3301 gggttacgcc gaacatgatc gactatttcg gacggccgta tgaaggcatc gccgtgttcg
     3361 acggcaaaaa gatcactgta acagggaccc tgtggaacgg caacaaaatt atcgacgagc
     3421 gcctgatcaa ccccgacggc tccctgctgt tccgagtaac catcaacgga gtgaccggct
     3481 ggcggctgtg cgaacgcatt ctggcgtaag gccgcgactc tagagtcgac ctgcaggcat
     3541 gcaagctgat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
     3601 tgagcaataa ctagcataac cccttggggc ggccgcttcg agcagacatg ataagataca
     3661 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     3721 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     3781 acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt tttttaagca
     3841 agtaaaacct ctacaaatgt ggtaaaatcg aattttaaca aaatattaac gcttacaatt
     3901 tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgcggat
     3961 ctgcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag gtaccttctg
     4021 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     4081 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     4141 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4201 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     4261 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     4321 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     4381 taattaactg ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag
     4441 gtgaggaact aaacccagga ggcagatcat gattgaacaa gatggattgc acgcaggttc
     4501 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     4561 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     4621 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     4681 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     4741 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4801 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4861 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4921 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4981 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     5041 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     5101 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     5161 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     5221 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     5281 gaaatgaccg accaagcgac gcccaacctg ccatcacgat ggccgcaata aaatatcttt
     5341 attttcatta catctgtgtg ttggtttttt gtgtgaatcg atagcgataa ggatcctctt
     5401 tgcgcttgcg ttttcccttg tccagatagc ccagtagctg acattcatcc ggggtcagca
     5461 ccgtttctgc ggactggctt tctacccggt atcagctcac tcaaaggcgg taatacggtt
     5521 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5581 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5641 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5701 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5761 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5821 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5881 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5941 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6001 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     6061 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6121 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6181 gcgcagaaaa aaaggatttc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6241 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6301 ctagatcctt ttatagtccg gaaatacagg aacgcacgct ggatggccct tcgctgggat
     6361 ggtgaaacca tgaaaaatgg cagcttcagt ggattaagtg ggggtaatgt ggcctgtacc
     6421 ctctggttgc ataggtattc atacggttaa aatttatcag gcgcgattgc ggcagttttt
     6481 cgggtggttt gttgccattt ttacctgtct gctgccgtga tcgcgctgaa cgcgttttag
     6541 cggtgcgtac aattaaggga ttatggtaaa tccacttact gtctgccctc gtagccatcg
     6601 agataaaccg cagtactccg gccacgatgc gtccggcgta gaggatcgag atct
//
