LOCUS       sequence-471861-        6103 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express NanoLuc(R)-MDM4 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 6103)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6103)
  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            "Unpublished"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: NanoLuc-MDM4 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6103
                     /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"
     primer_bind     complement(3182..3200)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3237..3358
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3248..3267)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3302..3321
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3603..3960
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3603..3794
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3626..3646)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3811..3946
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3975..4022
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4054..4848
                     /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(4108..4127)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4718..4737
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4912..4960
                     /note="synthetic polyadenylation signal"
     rep_origin      5117..5662
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     5563..5582
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5779..5796
                     /label=L4440
                     /note="L4440 vector, forward primer"
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 gacatcattt tccacctctg
     1621 ctcagtgttc aacatctgac agtgcttgca ggatctctcc tggacaaatc aatcaggtac
     1681 gaccaaaact gccgcttttg aagattttgc atgcagcagg tgcgcaaggt gaaatgttca
     1741 ctgttaaaga ggtcatgcac tatttaggtc agtacataat ggtgaagcaa ctttatgatc
     1801 agcaggagca gcatatggta tattgtggtg gagatctttt gggagaacta ctgggacgtc
     1861 agagcttctc cgtgaaagac ccaagccctc tctatgatat gctaagaaag aatcttgtca
     1921 ctttagccac tgctactaca gatgctgctc agactctcgc tctcgcacag gatcacagta
     1981 tggatattcc aagtcaagac caactgaagc aaagtgcaga ggaaagttcc acttccagaa
     2041 aaagaactac agaagacgat atccccacac tgcctacctc agagcataaa tgcatacatt
     2101 ctagagaaga tgaagactta attgaaaatt tagcccaaga tgaaacatct aggctggacc
     2161 ttggatttga ggagtgggat gtagctggcc tgccttggtg gtttttagga aacttgagaa
     2221 gcaactatac acctagaagt aatggctcaa ctgatttaca gacaaatcag gatgtgggta
     2281 ctgccattgt ttcagatact acagatgact tgtggttttt gaatgagtca gtatcagagc
     2341 agttaggtgt tggaataaaa gttgaagctg ctgatactga acaaacaagt gaagaagtag
     2401 ggaaagtaag tgacaaaaag gtgattgaag tgggaaaaaa tgatgacctg gaggactcta
     2461 agtccttaag tgatgatacc gatgtagagg ttacctctga ggatgagtgg cagtgtactg
     2521 aatgcaagaa atttaactct ccaagcaaga ggtactgttt tcgttgttgg gccttgagga
     2581 aggattggta ttcagattgt tcaaagttaa cccattctct ctccacgtct gatatcactg
     2641 ccatacctga aaaggaaaat gaaggaaatg atgtccctga ttgtcgaaga accatttcgg
     2701 ctcctgtcgt tagacctaaa gatgcgtata taaagaaaga aaactccaaa ctttttgatc
     2761 cctgcaactc agtggaattc ttggatttgg ctcacagttc tgaaagccaa gagaccatct
     2821 caagcatggg agaacagtta gataaccttt ctgaacagag aacagataca gaaaacatgg
     2881 aggattgcca gaatctcttg aagccatgta gcttatgtga gaaaagacca cgagacggga
     2941 acattattca tggaaggacg ggccatcttg tcacttgttt tcactgtgcc agaagactaa
     3001 agaaggctgg ggcttcatgc cctatttgca agaaagagat tcagctggtt attaaggttt
     3061 ttatagcagt ttaaacgaat tcgggctcgg tacccgggga tcctctagag tcgacctgca
     3121 ggcatgcaag ctgatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
     3181 accgctgagc aataactagc ataacccctt ggggcggccg cttcgagcag acatgataag
     3241 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     3301 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     3361 caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggttttttt
     3421 aagcaagtaa aacctctaca aatgtggtaa aatcgaattt taacaaaata ttaacgctta
     3481 caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     3541 cggatctgcg cagcaccatg gcctgaaata acctctgaaa gaggaacttg gttaggtacc
     3601 ttctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3661 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3721 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3781 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3841 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     3901 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3961 aagcttaatt aactgttgac aattaatcat cggcatagta tatcggcata gtataatacg
     4021 acaaggtgag gaactaaacc caggaggcag atcatgattg aacaagatgg attgcacgca
     4081 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     4141 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     4201 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     4261 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     4321 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     4381 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     4441 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     4501 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     4561 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     4621 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     4681 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     4741 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     4801 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     4861 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgatggccg caataaaata
     4921 tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagc gataaggatc
     4981 cgctcatgcg atgataagct gtcaaacatg agaattacaa cttatatcgt atggggctga
     5041 cttcaggtgc tacatttgaa gagataaatt gcactgaaat ctagaaatat tttatctgat
     5101 taataagatg atcttcttga gatcgttttg gtctgcgcgt aatctcttgc tctgaaaacg
     5161 aaaaaaccgc cttgcagggc ggtttttcga aggttctctg agctaccaac tctttgaacc
     5221 gaggtaactg gcttggagga gcgcagtcac caaaacttgt cctttcagtt tagccttaac
     5281 cggcgcatga cttcaagact aactcctcta aatcaattac cagtggctgc tgccagtggt
     5341 gcttttgcat gtctttccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5401 tcggactgaa cggggggttc gtgcatacag tccagcttgg agcgaactgc ctacccggaa
     5461 ctgagtgtca ggcgtggaat gagacaaacg cggccataac agcggaatga caccggtaaa
     5521 ccgaaaggca ggaacaggag agcgcacgag ggagccgcca gggggaaacg cctggtatct
     5581 ttatagtcct gtcgggtttc gccaccactg atttgagcgt cagatttcgt gatgcttgtc
     5641 aggggggcgg agcctatgga aaaacggctt tgccgcggcc ctctcacttc cctgttaagt
     5701 atcttcctgg catcttccag gaaatctccg ccccgttcgt aagccatttc cgctcgccgc
     5761 agtcgaacga ccgagcgtag cgagtcagtg agcgaggaag cggaatatat cctgtatcac
     5821 atattctgct gacgcaccgg tgcagccttt tttctcctgc cacatgaagc acttcactga
     5881 caccctcatc agtgccaaca tagtaagcca gtatacactc cgctagcgct gatgtccggc
     5941 ggtgcttttg ccgttacgca ccaccccgtc atgagattac gcgttttagc ggtgcgtaca
     6001 attaagggat tatggtaaat ccacttactg tctgccctcg tagccatcga gataaaccgc
     6061 agtactccgg ccacgatgcg tccggcgtag aggatcgaga tct
//
