LOCUS       sequence-479210-        5812 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express NanoLuc-MEK1 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 5812)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5812)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: NanoBRET Assay Vector, NanoLuc-MEK1
FEATURES             Location/Qualifiers
     source          1..5812
                     /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(2891..2909)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2946..3067
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2957..2976)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3011..3030
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3312..3669
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3312..3503
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3335..3355)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3520..3655
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3684..3731
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3763..4557
                     /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(3817..3836)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4427..4446
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4621..4669
                     /note="synthetic polyadenylation signal"
     rep_origin      4826..5371
                     /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     5272..5291
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5488..5505
                     /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 gcccaagaag aagccgacgc
     1621 ccatccagct gaacccggcc cccgacggct ctgcagttaa cgggaccagc tctgcggaga
     1681 ccaacttgga ggccttgcag aagaagctgg aggagctaga gcttgatgag cagcagcgaa
     1741 agcgccttga ggcctttctt acccagaagc agaaggtggg agaactgaag gatgacgact
     1801 ttgagaagat cagtgagctg ggggctggca atggcggtgt ggtgttcaag gtctcccaca
     1861 agccttctgg cctggtcatg gccagaaagc taattcatct ggagatcaaa cccgcaatcc
     1921 ggaaccagat cataagggag ctgcaggttc tgcatgagtg caactctccg tacatcgtgg
     1981 gcttctatgg tgcgttctac agcgatggcg agatcagtat ctgcatggag cacatggatg
     2041 gaggttctct ggatcaagtc ctgaagaaag ctggaagaat tcctgaacaa attttaggaa
     2101 aagttagcat tgctgtaata aaaggcctga catatctgag ggagaagcac aagatcatgc
     2161 acagagatgt caagccctcc aacatcctag tcaactcccg tggggagatc aagctctgtg
     2221 actttggggt cagcgggcag ctcatcgact ccatggccaa ctccttcgtg ggcacaaggt
     2281 cctacatgtc gccagaaaga ctccagggga ctcattactc tgtgcagtca gacatctgga
     2341 gcatgggact gtctctggta gagatggcgg ttgggaggta tcccatccct cctccagatg
     2401 ccaaggagct ggagctgatg tttgggtgcc aggtggaagg agatgcggct gagaccccac
     2461 ccaggccaag gacccccggg aggcccctta gctcatacgg aatggacagc cgacctccca
     2521 tggcaatttt tgagttgttg gattacatag tcaacgagcc tcctccaaaa ctgcccagtg
     2581 gagtgttcag tctggaattt caagattttg tgaataaatg cttaataaaa aaccccgcag
     2641 agagagcaga tttgaagcaa ctcatggttc atgcttttat caagagatct gatgctgagg
     2701 aagtggattt tgcaggttgg ctctgctcca ccatcggcct taaccagccc agcacaccaa
     2761 cccatgctgc tggcgtcgtt taaacgaatt cgggctcggt acccggggat cctctagagt
     2821 cgacctgcag gcatgcaagc tgatccggct gctaacaaag cccgaaagga agctgagttg
     2881 gctgctgcca ccgctgagca ataactagca taaccccttg gggcggccgc ttcgagcaga
     2941 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     3001 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     3061 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga
     3121 ggttttttta agcaagtaaa acctctacaa atgtggtaaa atcgaatttt aacaaaatat
     3181 taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     3241 ccgcatacgc ggatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg
     3301 ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     3361 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3421 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3481 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3541 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3601 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3661 ttttgcaaaa agcttaatta actgttgaca attaatcatc ggcatagtat atcggcatag
     3721 tataatacga caaggtgagg aactaaaccc aggaggcaga tcatgattga acaagatgga
     3781 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     3841 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     3901 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     3961 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4021 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4081 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4141 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4201 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4261 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     4321 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4381 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4441 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4501 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     4561 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgatggccgc
     4621 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagcg
     4681 ataaggatcc gctcatgcga tgataagctg tcaaacatga gaattacaac ttatatcgta
     4741 tggggctgac ttcaggtgct acatttgaag agataaattg cactgaaatc tagaaatatt
     4801 ttatctgatt aataagatga tcttcttgag atcgttttgg tctgcgcgta atctcttgct
     4861 ctgaaaacga aaaaaccgcc ttgcagggcg gtttttcgaa ggttctctga gctaccaact
     4921 ctttgaaccg aggtaactgg cttggaggag cgcagtcacc aaaacttgtc ctttcagttt
     4981 agccttaacc ggcgcatgac ttcaagacta actcctctaa atcaattacc agtggctgct
     5041 gccagtggtg cttttgcatg tctttccggg ttggactcaa gacgatagtt accggataag
     5101 gcgcagcggt cggactgaac ggggggttcg tgcatacagt ccagcttgga gcgaactgcc
     5161 tacccggaac tgagtgtcag gcgtggaatg agacaaacgc ggccataaca gcggaatgac
     5221 accggtaaac cgaaaggcag gaacaggaga gcgcacgagg gagccgccag ggggaaacgc
     5281 ctggtatctt tatagtcctg tcgggtttcg ccaccactga tttgagcgtc agatttcgtg
     5341 atgcttgtca ggggggcgga gcctatggaa aaacggcttt gccgcggccc tctcacttcc
     5401 ctgttaagta tcttcctggc atcttccagg aaatctccgc cccgttcgta agccatttcc
     5461 gctcgccgca gtcgaacgac cgagcgtagc gagtcagtga gcgaggaagc ggaatatatc
     5521 ctgtatcaca tattctgctg acgcaccggt gcagcctttt ttctcctgcc acatgaagca
     5581 cttcactgac accctcatca gtgccaacat agtaagccag tatacactcc gctagcgctg
     5641 atgtccggcg gtgcttttgc cgttacgcac caccccgtca tgagattacg cgttttagcg
     5701 gtgcgtacaa ttaagggatt atggtaaatc cacttactgt ctgccctcgt agccatcgag
     5761 ataaaccgca gtactccggc cacgatgcgt ccggcgtaga ggatcgagat ct
//
