LOCUS       sequence-473377-        7838 bp    DNA     circular SYN 06-JUN-2025
DEFINITION  Express NanoLuc(R)-PRMT5 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 7838)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 7838)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 6, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: NanoBRET Assay Vector, NanoLuc-PRMT5
FEATURES             Location/Qualifiers
     source          1..7838
                     /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"
     CDS             1065..1577
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     regulatory      1597..1606
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3606..3624)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3656..3777
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3667..3686)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3721..3740
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4022..4379
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4022..4213
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4045..4065)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4230..4365
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4410..5447
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5426..5434
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    5471..5519
                     /note="synthetic polyadenylation signal"
     promoter        5675..5779
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5780..5848,5849..6640)
                     /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     complement(5998..6017)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6798..7386)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6878..6897)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7502..7785
                     /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 ggctagcgct caccatggtc ttcacactcg
     1081 aagatttcgt tggggactgg cgacagacag ccggctacaa cctggaccaa gtccttgaac
     1141 agggaggtgt gtccagtttg tttcagaatc tcggggtgtc cgtaactccg atccaaagga
     1201 ttgtcctgag cggtgaaaat gggctgaaga tcgacatcca tgtcatcatc ccgtatgaag
     1261 gtctgagcgg cgaccaaatg ggccagatcg aaaaaatttt taaggtggtg taccctgtgg
     1321 atgatcatca ctttaaggtg atcctgcact atggcacact ggtaatcgac ggggttacgc
     1381 cgaacatgat cgactatttc ggacggccgt atgaaggcat cgccgtgttc gacggcaaaa
     1441 agatcactgt aacagggacc ctgtggaacg gcaacaaaat tatcgacgag cgcctgatca
     1501 accccgacgg ctccctgctg ttccgagtaa ccatcaacgg agtgaccggc tggcggctgt
     1561 gcgaacgcat tctggcgggc tcgagcggcg cgatcgccat ggcggcgatg gcggtcgggg
     1621 gtgctggtgg gagccgcgtg tccagcggga gggacctgaa ttgcgtcccc gaaatagctg
     1681 acacactagg ggctgtggcc aagcaggggt ttgatttcct ctgcatgcct gtcttccatc
     1741 cgcgtttcaa gagggagttc attcaggaac ctgctaagaa tcggcccggt ccccagacac
     1801 gatcagacct actgctgtca ggaagggact ggaatacgct aattgtggga aagctttctc
     1861 catggattcg tccagactca aaagtggaga agattcgcag gaactccgag gcggccatgt
     1921 tacaggagct gaattttggt gcatatttgg gtcttccagc tttcctgctg ccccttaatc
     1981 aggaagataa caccaacctg gccagagttt tgaccaacca catccacact ggccatcact
     2041 cttccatgtt ctggatgcgg gtacccttgg tggcaccaga ggacctgaga gatgatataa
     2101 ttgagaatgc accaactaca cacacagagg agtacagtgg ggaggagaaa acgtggatgt
     2161 ggtggcacaa cttccggact ttgtgtgact atagtaagag gattgcagtg gctcttgaaa
     2221 ttggggctga cctcccatct aatcatgtca ttgatcgctg gcttggggag cccatcaaag
     2281 cagccattct ccccactagc attttcctga ccaataagaa gggatttcct gttctttcta
     2341 agatgcacca gaggctcatc ttccggctcc tcaagttgga ggtgcagttc atcatcacag
     2401 gcaccaacca ccactcagag aaggagttct gctcctacct ccaatacctg gaatacttaa
     2461 gccagaaccg tcctccacct aatgcctatg aactctttgc caagggctat gaagactatc
     2521 tgcagtcccc gcttcagcca ctgatggaca atctggaatc tcagacatat gaagtgtttg
     2581 aaaaggaccc catcaaatac tctcagtacc agcaggccat ctataaatgt ctgctagacc
     2641 gagtaccaga agaggagaag gataccaatg tccaggtact gatggtgctg ggagcaggac
     2701 ggggacccct ggtgaacgct tccctgcggg cagccaagca ggccgaccgg cggataaagc
     2761 tgtatgctgt ggagaaaaac ccaaatgccg tggtgacgct agagaactgg cagtttgaag
     2821 aatggggaag ccaagtgacc gtagtctcat cagacatgag ggaatgggtg gctccagaga
     2881 aagcagacat cattgtcagt gagcttctgg gctcatttgc tgacaatgaa ttgtcgcctg
     2941 agtgcctgga tggagcccag cacttcctaa aagatgatgg tgtgagcatc cccggggagt
     3001 acacttcctt tctggctccc atctcttcct ccaagctgta caatgaggtc cgagcctgta
     3061 gggagaagga ccgtgaccct gaggcccagt ttgagatgcc ttatgtggta cggctgcaca
     3121 acttccacca gctctctgca ccccagccct gtttcacctt cagccatccc aacagagatc
     3181 ctatgattga caacaaccgc tattgcacct tggaatttcc tgtggaggtg aacacagtac
     3241 tacatggctt tgccggctac tttgagactg tgctttatca ggacatcact ctgagtatcc
     3301 gtccagagac tcactctcct gggatgttct catggtttcc catcctcttc cctattaagc
     3361 agcccataac ggtacgtgaa ggccaaacca tctgtgtgcg tttctggcga tgcagcaatt
     3421 ccaagaaggt gtggtatgag tgggctgtga cagcaccagt ctgttctgct attcataacc
     3481 ccacaggccg ctcatatacc attggcctcg tttaaacgaa ttcgggctcg gtacccgggg
     3541 atcctctaga tttgggccca attcctgcag gattttgcgg ccgcttgctg agttggctgc
     3601 tgccaccgct gagcaataac tagcataacc ccttggccgc ttcgagcaga catgataaga
     3661 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     3721 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     3781 aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttta
     3841 agcaagtaaa acctctacaa atgtggtaaa atcgaatttt aacaaaatat taacgcttac
     3901 aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgc
     3961 ggatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg ttaggtacct
     4021 tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     4081 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     4141 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4201 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4261 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     4321 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     4381 agctcgattc ttctgacact agcgccacca tgaagaagcc cgaactcacc gctaccagcg
     4441 ttgaaaaatt tctcatcgag aagttcgaca gtgtgagcga cctgatgcag ttgtcggagg
     4501 gcgaagagag ccgagccttc agcttcgatg tcggcggacg cggctatgta ctgcgggtga
     4561 atagctgcgc tgatggcttc tacaaagacc gctacgtgta ccgccacttc gccagcgctg
     4621 cactacccat ccccgaagtg ttggacatcg gcgagttcag cgagagcctg acatactgca
     4681 tcagtagacg cgcccaaggc gttactctcc aagacctccc cgaaacagag ctgcctgctg
     4741 tgttacagcc tgtcgccgaa gctatggatg ctattgccgc cgccgacctc agtcaaacca
     4801 gcggcttcgg cccattcggg ccccaaggca tcggccagta cacaacctgg cgggatttca
     4861 tttgcgccat tgctgatccc catgtctacc actggcagac cgtgatggac gacaccgtgt
     4921 ccgccagcgt agctcaagcc ctggacgaac tgatgctgtg ggccgaagac tgtcccgagg
     4981 tgcgccacct cgtccatgcc gacttcggca gcaacaacgt cctgaccgac aacggccgca
     5041 tcaccgccgt aatcgactgg tccgaagcta tgttcgggga cagtcagtac gaggtggcca
     5101 acatcttctt ctggcggccc tggctggctt gcatggagca gcagactcgc tacttcgagc
     5161 gccggcatcc cgagctggcc ggcagccctc gtctgcgagc ctacatgctg cgcatcggcc
     5221 tggatcagct ctaccagagc ctcgtggacg gcaacttcga cgatgctgcc tgggctcaag
     5281 gccgctgcga tgccatcgtc cgcagcgggg ccggcaccgt cggtcgcaca caaatcgctc
     5341 gccggagcgc agccgtatgg accgacggct gcgtcgaggt gctggccgac agcggcaacc
     5401 gccggcccag tacacgaccg cgcgctaagg aggtaggtcg agtttagact ctagccatca
     5461 cgatggccgc aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga
     5521 atcgatagcg ataaggatcc tctttgcgct tgcgttttcc cttgtccaga tagcccagta
     5581 gctgacattc atccggggtc agcaccgttt ctgcggactg gctttctacg taatggtttc
     5641 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     5701 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     5761 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     5821 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     5881 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     5941 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttca aagttctgct
     6001 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     6061 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     6121 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     6181 cttacttctg acaactatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     6241 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     6301 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     6361 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     6421 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     6481 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     6541 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     6601 gatcgctgag ataggtgcct cactgattaa gcattggtaa ttcgaaatga ccgaccaagc
     6661 gacgcccaac cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6721 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6781 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6841 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6901 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6961 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     7021 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     7081 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     7141 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     7201 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     7261 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     7321 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     7381 tttcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     7441 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttatag
     7501 tccggaaata caggaacgca cgctggatgg cccttcgctg ggatggtgaa accatgaaaa
     7561 atggcagctt cagtggatta agtgggggta atgtggcctg taccctctgg ttgcataggt
     7621 attcatacgg ttaaaattta tcaggcgcga ttgcggcagt ttttcgggtg gtttgttgcc
     7681 atttttacct gtctgctgcc gtgatcgcgc tgaacgcgtt ttagcggtgc gtacaattaa
     7741 gggattatgg taaatccact tactgtctgc cctcgtagcc atcgagataa accgcagtac
     7801 tccggccacg atgcgtccgg cgtagaggat cgagatct
//
