LOCUS       sequence-474732-        5068 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express NanoLuc(R)-TOM20 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 5068)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5068)
  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            "Unpublished"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-TOM20 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5068
                     /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(2147..2165)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2202..2323
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2213..2232)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2267..2286
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2568..2925
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2568..2759
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2591..2611)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2776..2911
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        2940..2987
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3019..3813
                     /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(3073..3092)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3683..3702
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3877..3925
                     /note="synthetic polyadenylation signal"
     rep_origin      4082..4627
                     /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     4528..4547
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4744..4761
                     /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 ggttggtcgt aattcagcta
     1621 tagcagcagg agtttgtggt gcactcttta taggatactg catatacttt gatcgcaaac
     1681 ggcggtcaga tccgaatttt aaaaaccgct tgcgggagcg cagaaagaag cagaaactgg
     1741 caaaagaaag agccggtctg agtaagctcc cggaccttaa agacgctgag gcggttcaaa
     1801 aatttttttt ggaggagatc cagctcgggg aagaactgtt ggcccaaggt gaatatgaga
     1861 aaggtgttga ccatttgact aacgcaatag cggtctgcgg tcaaccgcag cagcttttgc
     1921 aagtattgca gcagaccctg cctccacctg tctttcagat gctgcttaca aagttgccta
     1981 ctataagtca gcgaattgtt agtgctcaat cattagcaga agatgatgtt gaagtttaaa
     2041 cgaattcggg ctcggtaccc ggggatcctc tagagtcgac ctgcaggcat gcaagctgat
     2101 ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
     2161 ctagcataac cccttggggc ggccgcttcg agcagacatg ataagataca ttgatgagtt
     2221 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     2281 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     2341 tcattttatg tttcaggttc agggggagat gtgggaggtt tttttaagca agtaaaacct
     2401 ctacaaatgt ggtaaaatcg aattttaaca aaatattaac gcttacaatt tcctgatgcg
     2461 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgcggat ctgcgcagca
     2521 ccatggcctg aaataacctc tgaaagagga acttggttag gtaccttctg aggcggaaag
     2581 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     2641 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     2701 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     2761 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     2821 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     2881 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct taattaactg
     2941 ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact
     3001 aaacccagga ggcagatcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     3061 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     3121 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     3181 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     3241 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     3301 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     3361 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     3421 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     3481 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     3541 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     3601 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     3661 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     3721 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     3781 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     3841 accaagcgac gcccaacctg ccatcacgat ggccgcaata aaatatcttt attttcatta
     3901 catctgtgtg ttggtttttt gtgtgaatcg atagcgataa ggatccgctc atgcgatgat
     3961 aagctgtcaa acatgagaat tacaacttat atcgtatggg gctgacttca ggtgctacat
     4021 ttgaagagat aaattgcact gaaatctaga aatattttat ctgattaata agatgatctt
     4081 cttgagatcg ttttggtctg cgcgtaatct cttgctctga aaacgaaaaa accgccttgc
     4141 agggcggttt ttcgaaggtt ctctgagcta ccaactcttt gaaccgaggt aactggcttg
     4201 gaggagcgca gtcaccaaaa cttgtccttt cagtttagcc ttaaccggcg catgacttca
     4261 agactaactc ctctaaatca attaccagtg gctgctgcca gtggtgcttt tgcatgtctt
     4321 tccgggttgg actcaagacg atagttaccg gataaggcgc agcggtcgga ctgaacgggg
     4381 ggttcgtgca tacagtccag cttggagcga actgcctacc cggaactgag tgtcaggcgt
     4441 ggaatgagac aaacgcggcc ataacagcgg aatgacaccg gtaaaccgaa aggcaggaac
     4501 aggagagcgc acgagggagc cgccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4561 gtttcgccac cactgatttg agcgtcagat ttcgtgatgc ttgtcagggg ggcggagcct
     4621 atggaaaaac ggctttgccg cggccctctc acttccctgt taagtatctt cctggcatct
     4681 tccaggaaat ctccgccccg ttcgtaagcc atttccgctc gccgcagtcg aacgaccgag
     4741 cgtagcgagt cagtgagcga ggaagcggaa tatatcctgt atcacatatt ctgctgacgc
     4801 accggtgcag ccttttttct cctgccacat gaagcacttc actgacaccc tcatcagtgc
     4861 caacatagta agccagtata cactccgcta gcgctgatgt ccggcggtgc ttttgccgtt
     4921 acgcaccacc ccgtcatgag attacgcgtt ttagcggtgc gtacaattaa gggattatgg
     4981 taaatccact tactgtctgc cctcgtagcc atcgagataa accgcagtac tccggccacg
     5041 atgcgtccgg cgtagaggat cgagatct
//
