LOCUS       sequence-471914-        5042 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express TOM20-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 5042)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5042)
  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            "Unpublished"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: TOM20-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5042
                     /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             1521..2033
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(2121..2139)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2176..2297
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2187..2206)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2241..2260
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2542..2899
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2542..2733
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2565..2585)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2750..2885
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        2914..2961
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2993..3787
                     /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(3047..3066)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3657..3676
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3851..3899
                     /note="synthetic polyadenylation signal"
     rep_origin      4056..4601
                     /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     4502..4521
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4718..4735
                     /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 ggctagcgat cgccatggtt gggcgcaatt
     1081 cagcgatagc ggcgggggtt tgcggggcgc tctttatagg atactgcata tactttgatc
     1141 gcaaacggcg gtcagatccg aattttaaaa accgcttgcg ggagcgcaga aagaagcaga
     1201 aactggcaaa agaaagagcc ggtctgagta agctcccgga ccttaaagac gctgaggcgg
     1261 ttcaaaaatt ttttttggag gagatccagc tcggggaaga actgttggcc caaggtgaat
     1321 atgagaaagg tgttgaccat ttgactaacg caatagcggt ctgcggtcaa ccgcagcagc
     1381 ttttgcaagt attgcagcag accctgcctc cacctgtctt tcagatgctg cttacaaagt
     1441 tgcctactat aagtcagcga attgtcagcg cccagtcatt ggcagaggac gatgtggagg
     1501 tttctctcgg ctcgagcggc gtcttcacac tcgaagattt cgttggggac tggcgacaga
     1561 cagccggcta caacctggac caagtccttg aacagggagg tgtgtccagt ttgtttcaga
     1621 atctcggggt gtccgtaact ccgatccaaa ggattgtcct gagcggtgaa aatgggctga
     1681 agatcgacat ccatgtcatc atcccgtatg aaggtctgag cggcgaccaa atgggccaga
     1741 tcgaaaaaat ttttaaggtg gtgtaccctg tggatgatca tcactttaag gtgatcctgc
     1801 actatggcac actggtaatc gacggggtta cgccgaacat gatcgactat ttcggacggc
     1861 cgtatgaagg catcgccgtg ttcgacggca aaaagatcac tgtaacaggg accctgtgga
     1921 acggcaacaa aattatcgac gagcgcctga tcaaccccga cggctccctg ctgttccgag
     1981 taaccatcaa cggagtgacc ggctggcggc tgtgcgaacg cattctggcg taaggccgcg
     2041 actctagagt cgacctgcag gcatgcaagc tgatccggct gctaacaaag cccgaaagga
     2101 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcggccgc
     2161 ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     2221 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     2281 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     2341 agatgtggga ggttttttta agcaagtaaa acctctacaa atgtggtaaa atcgaatttt
     2401 aacaaaatat taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg
     2461 gtatttcaca ccgcatacgc ggatctgcgc agcaccatgg cctgaaataa cctctgaaag
     2521 aggaacttgg ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     2581 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     2641 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2701 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     2761 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     2821 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     2881 aggcctaggc ttttgcaaaa agcttaatta actgttgaca attaatcatc ggcatagtat
     2941 atcggcatag tataatacga caaggtgagg aactaaaccc aggaggcaga tcatgattga
     3001 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     3061 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     3121 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     3181 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     3241 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     3301 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     3361 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     3421 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     3481 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     3541 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     3601 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     3661 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     3721 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     3781 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     3841 cgatggccgc aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga
     3901 atcgatagcg ataaggatcc gctcatgcga tgataagctg tcaaacatga gaattacaac
     3961 ttatatcgta tggggctgac ttcaggtgct acatttgaag agataaattg cactgaaatc
     4021 tagaaatatt ttatctgatt aataagatga tcttcttgag atcgttttgg tctgcgcgta
     4081 atctcttgct ctgaaaacga aaaaaccgcc ttgcagggcg gtttttcgaa ggttctctga
     4141 gctaccaact ctttgaaccg aggtaactgg cttggaggag cgcagtcacc aaaacttgtc
     4201 ctttcagttt agccttaacc ggcgcatgac ttcaagacta actcctctaa atcaattacc
     4261 agtggctgct gccagtggtg cttttgcatg tctttccggg ttggactcaa gacgatagtt
     4321 accggataag gcgcagcggt cggactgaac ggggggttcg tgcatacagt ccagcttgga
     4381 gcgaactgcc tacccggaac tgagtgtcag gcgtggaatg agacaaacgc ggccataaca
     4441 gcggaatgac accggtaaac cgaaaggcag gaacaggaga gcgcacgagg gagccgccag
     4501 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccaccactga tttgagcgtc
     4561 agatttcgtg atgcttgtca ggggggcgga gcctatggaa aaacggcttt gccgcggccc
     4621 tctcacttcc ctgttaagta tcttcctggc atcttccagg aaatctccgc cccgttcgta
     4681 agccatttcc gctcgccgca gtcgaacgac cgagcgtagc gagtcagtga gcgaggaagc
     4741 ggaatatatc ctgtatcaca tattctgctg acgcaccggt gcagcctttt ttctcctgcc
     4801 acatgaagca cttcactgac accctcatca gtgccaacat agtaagccag tatacactcc
     4861 gctagcgctg atgtccggcg gtgcttttgc cgttacgcac caccccgtca tgagattacg
     4921 cgttttagcg gtgcgtacaa ttaagggatt atggtaaatc cacttactgt ctgccctcgt
     4981 agccatcgag ataaaccgca gtactccggc cacgatgcgt ccggcgtaga ggatcgagat
     5041 ct
//
