LOCUS       sequence-474576-        5545 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express GR-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 5545)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5545)
  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            "Unpublished"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: NanoBRET Assay Acceptor Vector, GR
FEATURES             Location/Qualifiers
     source          1..5545
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(50..253)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(79..99)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        254..633
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    824..1107
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     rep_origin      1223..1811
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1712..1731
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1983..2777)
                     /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="MLEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2094..2113)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2704..2723
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     polyA_signal    3176..3297
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3213..3232)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3267..3286
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3334..3352
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(3440..4324)
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     CDS             complement(4340..4360)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     primer_bind     complement(5264..5283)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5265..5283)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5325..5344)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          5327..5459
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
ORIGIN
        1 gataaactac cgcaataaag cttctagtga tctgacggtt cactaaacca gctctgctta
       61 tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt caacggggcg gggttattac
      121 gacattttgg aaagtcccgt tgattttggt gccaaaacaa actcccattg acgtcaatgg
      181 ggtggagact tggaaatccc cgtgagtcaa accgctatcc acgcccattg gtgtactgcc
      241 aaaaccgcat caccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa
      301 agtcccgtaa ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt
      361 caataggggg cggacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc
      421 gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat
      481 acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta
      541 ccgtaagtta tgtaacgcgg aactccatat atgggctatg aactaatgac cccgtaattg
      601 attactatta ataactagtc aataatcaat gccaacatgg cggtcatatt ggacatgagc
      661 caatataaat gtacatatta tgatatagat acaacgtatg caatggccaa tagccaatat
      721 tgatttatgc tatataacca atgaataata tggctaatgg ccaatattga agatctcgat
      781 cctctacgcc ggacgcatcg tggccggagt actgcggttt atctcgatgg ctacgagggc
      841 agacagtaag tggatttacc ataatccctt aattgtacgc accgctaaaa cgcgttcagc
      901 gcgatcacgg cagcagacag gtaaaaatgg caacaaacca cccgaaaaac tgccgcaatc
      961 gcgcctgata aattttaacc gtatgaatac ctatgcaacc agagggtaca ggccacatta
     1021 cccccactta atccactgaa gctgccattt ttcatggttt caccatccca gcgaagggcc
     1081 atccagcgtg cgttcctgta tttccggact ataaaaggat ctaggtgaag atcctttttg
     1141 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     1201 tagaaaagat caaaggatct tcttgaaatc ctttttttct gcgcgtaatc tgctgcttgc
     1261 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     1321 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     1381 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     1441 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     1501 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     1561 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     1621 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     1681 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     1741 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     1801 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     1861 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     1921 ttgagtgagc tgataccggt tgggcgtcgc ttggtcggtc atttcgaacc ccagagtccc
     1981 gctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg
     2041 ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca
     2101 cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg
     2161 aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc gccatgagtc
     2221 acgacgagat cctcaccatc cggcatacgc gccttgagcc tggcgaacag ttcggctggc
     2281 gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga
     2341 gtgcgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca
     2401 agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg
     2461 tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct
     2521 tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc
     2581 cgcgctgcct cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga
     2641 accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt
     2701 tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat
     2761 ccatcttgtt caagcatgcg aaacgaccgt catcctgtct cttgatcaga tcttgatccc
     2821 ctgcgccatc agatccttgg cggcaagaaa gccatccagt ttactttgca gggcttccca
     2881 accttaccag agggcgcccc agctggcaat tcttttgaag ctcacgctgc cgcaagcact
     2941 cagggcgcaa gggctgctaa aggaagcgga acacgtagaa agccagtccg cagaaacggt
     3001 gctgaccccg gatgaatgtc agctactggg ctatctggac aagggaaaac gcaagcgcaa
     3061 agaggatcca ttagaattcg attttaccac atttgtagag gttttacttg cttaaaaaaa
     3121 cctcccacat ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     3181 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     3241 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     3301 tgtctgctcg aagcggccgc cccaaggggt tatgctagtt attgctcagc ggtggcagca
     3361 gccaactcag cttcctttcg ggctttgtta gcagccggat cagcttgcat gcctgcaggt
     3421 cgactctaga attctattaa ccggaaatct ccagagtaga cagccagcgc gcgatctcgc
     3481 tgccgatcag gtccgggttg tcttcttgca gcagattcag acccgggccg atgtccacag
     3541 ccttgcagtt aggcaggctt ttggccaggc gagcggcttc ggccggtggg atcagaacgc
     3601 ctggggtgcc ccagaacagc agcttcggga caggggactg gtgcagccag tccatgtatt
     3661 cttcgaccag cgcgacgatg ttcgctggct caccggcgat tggcagctcg tttgggaagc
     3721 gccacagtgg ctcgcggtca acaggattca ggaacggctc gcggtaatgg tccatctcga
     3781 cttcagtcag cgggcggacg acacccatcg gcagcgtacc ctcgataaaa acgttctgat
     3841 cgatgatcag cttgcggccg acgtcggtgg tgcggaaggc ctggaaggtc tcgcgggcaa
     3901 attctggcca ttcgtcccag gtcgggatag ggcggatgaa ctccataaat gcaatacctt
     3961 tgacgcgctc tggattgcgc ttggcccagt ggaaacccag agcggagccc cagtcgtgaa
     4021 tgaccaggac gacctcttcc agacccaggg cttcgatgaa ggcatccatg aagcggacgt
     4081 ggtcgtcgaa gaaataaccc aggtctggtt tgtcggattt gcccataccg atcaggtctg
     4141 gagcaatgca gcgatgggtc ggtgcaacat gcgggatgat gttgcgccac acgtaggagg
     4201 aggtcgggtt accgtgcagg aacagcacag gggtgccatc gcgcggacca acatcgacgt
     4261 agtgcatgcg ctcgcccagg acttccacat aatgggggtc gaatggaaag ccagtaccga
     4321 tttcggatcc atcgttatcg ctctgaaagt acagatcctc agtggttggc tcgagagaaa
     4381 ccgtctcctc caaatgtgta tcactttgct tctttgagtt tgtatcttgg atgccacatt
     4441 ttttcacatc catcattctc ccttttctta aacggaagat gaatgtcagt gctacaccaa
     4501 ggcataataa gatggctccc agaattacca agtgagtcct ttcatttgga ggatgtgcca
     4561 gaggtagttc tgggatgacc aattcagctg tatggttttc ctcaggatct aatctcctaa
     4621 aagtgcagta gaaaatctca ttagttgttg tgttgattct cagtgtgctg gtcacattga
     4681 aaagcttctc ctctctcttg gaattggtgg tggtggtctt accactcagg acttgatggt
     4741 cactgcttgt ccagatgact tcggccttgg ggtagccctc agcctgacat gtcagttcat
     4801 gttcagaggt gactggatcc acaaccaaaa ttctttggtt gattttgttg tatggggcat
     4861 tgactttcac agtaattcgc ttgtagtcgg caccaccata gctgatcatg cagcggtaca
     4921 cccctgcatc ctgcaatttc acatctgtga tctgaagtgc agcatttccc agggagagct
     4981 ggtccttcaa cagccgggcc ctctgtctgt agctactatg ctgaaccttc aggtcttcct
     5041 ctccatgcac aaattgaatg atgttcttat cctccatttc ccaatagaca attagtgcag
     5101 ccaggtctaa ttgtttttct actgggaatt tgcattcaat tgtcatattg ctaccatact
     5161 ctaccacata taggtccttg ggaaccgtga cagtaaatgc gttcagcaaa tgccagtagg
     5221 tcatgaatat aaagacagca aatatcctca tggcgatcgc tagccctata gtgagtcgta
     5281 ttaatactct agccttaaga gctgtaattg aactgggagt ggacacctgt ggagagaaag
     5341 gcaaagtgga tgtcagtaag accaataggt gcctatcaga aacgcaagag tcttctctgt
     5401 ctcgacaagc ccagtttcta ttggtctcct taaacctgtc ttgtaacctt gatacttacc
     5461 tgcccagtgc ctcacgacca acttctgcag cttaagttcg agactgttgt gtcagaagca
     5521 ctgactgcgt tagcaattta actgt
//
