LOCUS       sequence-478459-        5786 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express MEK1-NanoLuc 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 5786)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5786)
  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            "Unpublished"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, MEK1-NanoLuc
FEATURES             Location/Qualifiers
     source          1..5786
                     /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             2265..2777
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(2865..2883)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2920..3041
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2931..2950)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2985..3004
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3286..3643
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3286..3477
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3309..3329)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3494..3629
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3658..3705
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3737..4531
                     /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(3791..3810)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4401..4420
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4595..4643
                     /note="synthetic polyadenylation signal"
     rep_origin      4800..5345
                     /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     5246..5265
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5462..5479
                     /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 cgccatgccc aagaagaagc
     1081 cgacgcccat ccagctgaac ccggcccccg acggctctgc agttaacggg accagctctg
     1141 cggagaccaa cttggaggcc ttgcagaaga agctggagga gctagagctt gatgagcagc
     1201 agcgaaagcg ccttgaggcc tttcttaccc agaagcagaa ggtgggagaa ctgaaggatg
     1261 acgactttga gaagatcagt gagctggggg ctggcaatgg cggtgtggtg ttcaaggtct
     1321 cccacaagcc ttctggcctg gtcatggcca gaaagctaat tcatctggag atcaaacccg
     1381 caatccggaa ccagatcata agggagctgc aggttctgca tgagtgcaac tctccgtaca
     1441 tcgtgggctt ctatggtgcg ttctacagcg atggcgagat cagtatctgc atggagcaca
     1501 tggatggagg ttctctggat caagtcctga agaaagctgg aagaattcct gaacaaattt
     1561 taggaaaagt tagcattgct gtaataaaag gcctgacata tctgagggag aagcacaaga
     1621 tcatgcacag agatgtcaag ccctccaaca tcctagtcaa ctcccgtggg gagatcaagc
     1681 tctgtgactt tggggtcagc gggcagctca tcgactccat ggccaactcc ttcgtgggca
     1741 caaggtccta catgtcgcca gaaagactcc aggggactca ttactctgtg cagtcagaca
     1801 tctggagcat gggactgtct ctggtagaga tggcggttgg gaggtatccc atccctcctc
     1861 cagatgccaa ggagctggag ctgatgtttg ggtgccaggt ggaaggagat gcggctgaga
     1921 ccccacccag gccaaggacc cccgggaggc cccttagctc atacggaatg gacagccgac
     1981 ctcccatggc aatttttgag ttgttggatt acatagtcaa cgagcctcct ccaaaactgc
     2041 ccagtggagt gttcagtctg gaatttcaag attttgtgaa taaatgctta ataaaaaacc
     2101 ccgcagagag agcagatttg aagcaactca tggttcatgc ttttatcaag agatctgatg
     2161 ctgaggaagt ggattttgca ggttggctct gctccaccat cggccttaac cagcccagca
     2221 caccaaccca tgctgctggc gtcgtttctc tcggctcgag cggcgtcttc acactcgaag
     2281 atttcgttgg ggactggcga cagacagccg gctacaacct ggaccaagtc cttgaacagg
     2341 gaggtgtgtc cagtttgttt cagaatctcg gggtgtccgt aactccgatc caaaggattg
     2401 tcctgagcgg tgaaaatggg ctgaagatcg acatccatgt catcatcccg tatgaaggtc
     2461 tgagcggcga ccaaatgggc cagatcgaaa aaatttttaa ggtggtgtac cctgtggatg
     2521 atcatcactt taaggtgatc ctgcactatg gcacactggt aatcgacggg gttacgccga
     2581 acatgatcga ctatttcgga cggccgtatg aaggcatcgc cgtgttcgac ggcaaaaaga
     2641 tcactgtaac agggaccctg tggaacggca acaaaattat cgacgagcgc ctgatcaacc
     2701 ccgacggctc cctgctgttc cgagtaacca tcaacggagt gaccggctgg cggctgtgcg
     2761 aacgcattct ggcgtaaggc cgcgactcta gagtcgacct gcaggcatgc aagctgatcc
     2821 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
     2881 agcataaccc cttggggcgg ccgcttcgag cagacatgat aagatacatt gatgagtttg
     2941 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     3001 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     3061 attttatgtt tcaggttcag ggggagatgt gggaggtttt tttaagcaag taaaacctct
     3121 acaaatgtgg taaaatcgaa ttttaacaaa atattaacgc ttacaatttc ctgatgcggt
     3181 attttctcct tacgcatctg tgcggtattt cacaccgcat acgcggatct gcgcagcacc
     3241 atggcctgaa ataacctctg aaagaggaac ttggttaggt accttctgag gcggaaagaa
     3301 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     3361 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     3421 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     3481 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3541 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     3601 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctta attaactgtt
     3661 gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa
     3721 acccaggagg cagatcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     3781 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     3841 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     3901 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     3961 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     4021 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     4081 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     4141 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     4201 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     4261 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     4321 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     4381 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     4441 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     4501 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     4561 caagcgacgc ccaacctgcc atcacgatgg ccgcaataaa atatctttat tttcattaca
     4621 tctgtgtgtt ggttttttgt gtgaatcgat agcgataagg atccgctcat gcgatgataa
     4681 gctgtcaaac atgagaatta caacttatat cgtatggggc tgacttcagg tgctacattt
     4741 gaagagataa attgcactga aatctagaaa tattttatct gattaataag atgatcttct
     4801 tgagatcgtt ttggtctgcg cgtaatctct tgctctgaaa acgaaaaaac cgccttgcag
     4861 ggcggttttt cgaaggttct ctgagctacc aactctttga accgaggtaa ctggcttgga
     4921 ggagcgcagt caccaaaact tgtcctttca gtttagcctt aaccggcgca tgacttcaag
     4981 actaactcct ctaaatcaat taccagtggc tgctgccagt ggtgcttttg catgtctttc
     5041 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcggact gaacgggggg
     5101 ttcgtgcata cagtccagct tggagcgaac tgcctacccg gaactgagtg tcaggcgtgg
     5161 aatgagacaa acgcggccat aacagcggaa tgacaccggt aaaccgaaag gcaggaacag
     5221 gagagcgcac gagggagccg ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5281 ttcgccacca ctgatttgag cgtcagattt cgtgatgctt gtcagggggg cggagcctat
     5341 ggaaaaacgg ctttgccgcg gccctctcac ttccctgtta agtatcttcc tggcatcttc
     5401 caggaaatct ccgccccgtt cgtaagccat ttccgctcgc cgcagtcgaa cgaccgagcg
     5461 tagcgagtca gtgagcgagg aagcggaata tatcctgtat cacatattct gctgacgcac
     5521 cggtgcagcc ttttttctcc tgccacatga agcacttcac tgacaccctc atcagtgcca
     5581 acatagtaag ccagtataca ctccgctagc gctgatgtcc ggcggtgctt ttgccgttac
     5641 gcaccacccc gtcatgagat tacgcgtttt agcggtgcgt acaattaagg gattatggta
     5701 aatccactta ctgtctgccc tcgtagccat cgagataaac cgcagtactc cggccacgat
     5761 gcgtccggcg tagaggatcg agatct
//
