LOCUS       sequence-473342-        5256 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express MBP-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 5256)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5256)
  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            "Unpublished"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: MBP-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5256
                     /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             1599..2111
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(2199..2217)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2254..2375
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2265..2284)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2319..2338
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2620..2977
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2620..2811
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2643..2663)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2828..2963
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        2992..3039
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3071..3865
                     /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(3125..3144)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3735..3754
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3929..3977
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4216..4804)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4296..4315)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4920..5203
                     /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 tcacagaaga
     1081 gaccctccca gaggcacgga tccaagtacc tggccacagc aagtaccatg gaccatgcca
     1141 ggcatggctt cctcccaagg cacagagaca cgggcatcct tgactccatc gggcgcttct
     1201 ttggcggtga caggggtgcg cccaagcggg gctctggcaa ggactcacac cacccggcaa
     1261 gaactgctca ctacggctcc ctgccccaga agtcacacgg ccggacccaa gatgaaaacc
     1321 ccgtagtcca cttcttcaag aacattgtga cgcctcgcac accacccccg tcgcagggaa
     1381 aggggagagg actgtccctg agcagattta gctggggggc cgaaggccag agaccaggat
     1441 ttggctacgg aggcagagcg tccgactata aatcggctca caagggattc aagggagtcg
     1501 atgcccaggg cacgctttcc aaaattttta agctgggagg aagagatagt cgctctggat
     1561 cacccatggc tagacgcgtt tctctcggct cgagcggcgt cttcacactc gaagatttcg
     1621 ttggggactg gcgacagaca gccggctaca acctggacca agtccttgaa cagggaggtg
     1681 tgtccagttt gtttcagaat ctcggggtgt ccgtaactcc gatccaaagg attgtcctga
     1741 gcggtgaaaa tgggctgaag atcgacatcc atgtcatcat cccgtatgaa ggtctgagcg
     1801 gcgaccaaat gggccagatc gaaaaaattt ttaaggtggt gtaccctgtg gatgatcatc
     1861 actttaaggt gatcctgcac tatggcacac tggtaatcga cggggttacg ccgaacatga
     1921 tcgactattt cggacggccg tatgaaggca tcgccgtgtt cgacggcaaa aagatcactg
     1981 taacagggac cctgtggaac ggcaacaaaa ttatcgacga gcgcctgatc aaccccgacg
     2041 gctccctgct gttccgagta accatcaacg gagtgaccgg ctggcggctg tgcgaacgca
     2101 ttctggcgta aggccgcgac tctagagtcg acctgcaggc atgcaagctg atccggctgc
     2161 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata
     2221 accccttggg gcggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa
     2281 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     2341 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     2401 tgtttcaggt tcagggggag atgtgggagg tttttttaag caagtaaaac ctctacaaat
     2461 gtggtaaaat cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc
     2521 tccttacgca tctgtgcggt atttcacacc gcatacgcgg atctgcgcag caccatggcc
     2581 tgaaataacc tctgaaagag gaacttggtt aggtaccttc tgaggcggaa agaaccagct
     2641 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     2701 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     2761 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     2821 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     2881 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     2941 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag cttaattaac tgttgacaat
     3001 taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa ctaaacccag
     3061 gaggcagatc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     3121 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     3181 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     3241 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     3301 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     3361 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     3421 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     3481 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     3541 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     3601 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     3661 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     3721 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     3781 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     3841 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     3901 acgcccaacc tgccatcacg atggccgcaa taaaatatct ttattttcat tacatctgtg
     3961 tgttggtttt ttgtgtgaat cgatagcgat aaggatcctc tttgcgcttg cgttttccct
     4021 tgtccagata gcccagtagc tgacattcat ccggggtcag caccgtttct gcggactggc
     4081 tttctacccg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4141 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4201 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4261 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4321 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4381 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4441 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4501 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4561 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4621 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     4681 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4741 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatt
     4801 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4861 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttatagtc
     4921 cggaaataca ggaacgcacg ctggatggcc cttcgctggg atggtgaaac catgaaaaat
     4981 ggcagcttca gtggattaag tgggggtaat gtggcctgta ccctctggtt gcataggtat
     5041 tcatacggtt aaaatttatc aggcgcgatt gcggcagttt ttcgggtggt ttgttgccat
     5101 ttttacctgt ctgctgccgt gatcgcgctg aacgcgtttt agcggtgcgt acaattaagg
     5161 gattatggta aatccactta ctgtctgccc tcgtagccat cgagataaac cgcagtactc
     5221 cggccacgat gcgtccggcg tagaggatcg agatct
//
