LOCUS       sequence-472943-        4984 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express WRN-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 4984)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4984)
  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            "Unpublished"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: WRN-NanoLuc pFC32K Vector
FEATURES             Location/Qualifiers
     source          1..4984
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(243..262)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(244..262)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(304..323)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          306..438
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(574..777)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(603..623)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        778..1157
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    1348..1631
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     rep_origin      1747..2335
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2236..2255
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2507..3301)
                     /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(2618..2637)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3228..3247
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     polyA_signal    3700..3821
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3737..3756)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3791..3810
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3858..3876
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(3964..4848)
                     /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(4864..4884)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
ORIGIN
        1 gcgtgctcgg tgtaggtgac tgcgtcccga atcacattct ccaggaacac cttcagcaca
       61 ccgcgggtct cctcgtaaat gaggccagag atccgcttaa cgccgccacg ccgagctaga
      121 cgccgaatgg caggcttggt gatgccctga atgttgtctc tcaagacctt gcggtggcgc
      181 ttagcgcccc ctttgcctaa gccttttccg ccctttcctc tgccggacat ggcgatcgct
      241 agccctatag tgagtcgtat taatactcta gccttaagag ctgtaattga actgggagtg
      301 gacacctgtg gagagaaagg caaagtggat gtcagtaaga ccaataggtg cctatcagaa
      361 acgcaagagt cttctctgtc tcgacaagcc cagtttctat tggtctcctt aaacctgtct
      421 tgtaaccttg atacttacct gcccagtgcc tcacgaccaa cttctgcagc ttaagttcga
      481 gactgttgtg tcagaagcac tgactgcgtt agcaatttaa ctgtgataaa ctaccgcaat
      541 aaagcttcta gtgatctgac ggttcactaa accagctctg cttatataga cctcccaccg
      601 tacacgccta ccgcccattt gcgtcaacgg ggcggggtta ttacgacatt ttggaaagtc
      661 ccgttgattt tggtgccaaa acaaactccc attgacgtca atggggtgga gacttggaaa
      721 tccccgtgag tcaaaccgct atccacgccc attggtgtac tgccaaaacc gcatcaccat
      781 ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc gtaaggtcat
      841 gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag ggggcggact
      901 tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat actccaccca
      961 ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca ttattgacgt
     1021 caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac
     1081 gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact attaataact
     1141 agtcaataat caatgccaac atggcggtca tattggacat gagccaatat aaatgtacat
     1201 attatgatat agatacaacg tatgcaatgg ccaatagcca atattgattt atgctatata
     1261 accaatgaat aatatggcta atggccaata ttgaagatct cgatcctcta cgccggacgc
     1321 atcgtggccg gagtactgcg gtttatctcg atggctacga gggcagacag taagtggatt
     1381 taccataatc ccttaattgt acgcaccgct aaaacgcgtt cagcgcgatc acggcagcag
     1441 acaggtaaaa atggcaacaa accacccgaa aaactgccgc aatcgcgcct gataaatttt
     1501 aaccgtatga atacctatgc aaccagaggg tacaggccac attaccccca cttaatccac
     1561 tgaagctgcc atttttcatg gtttcaccat cccagcgaag ggccatccag cgtgcgttcc
     1621 tgtatttccg gactataaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     1681 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     1741 atcttcttga aatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     1801 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     1861 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     1921 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     1981 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     2041 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     2101 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     2161 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     2221 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     2281 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     2341 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     2401 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     2461 cggttgggcg tcgcttggtc ggtcatttcg aaccccagag tcccgctcag aagaactcgt
     2521 caagaaggcg atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga
     2581 ggaagcggtc agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgcta
     2641 tgtcctgata gcggtccgcc acacccagcc ggccacagtc gatgaatcca gaaaagcggc
     2701 cattttccac catgatattc ggcaagcagg catcgccatg agtcacgacg agatcctcac
     2761 catccggcat acgcgccttg agcctggcga acagttcggc tggcgcgagc ccctgatgct
     2821 cttcgtccag atcatcctga tcgacaagac cggcttccat ccgagtgcgt gctcgctcga
     2881 tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc
     2941 gcattgcatc agccatgatg gatactttct cggcaggagc aaggtgagat gacaggagat
     3001 cctgccccgg cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga
     3061 gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtcct
     3121 gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg
     3181 ctgacagccg gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc
     3241 cgaatagcct ctccacccaa gcggccggag aacctgcgtg caatccatct tgttcaagca
     3301 tgcgaaacga ccgtcatcct gtctcttgat cagatcttga tcccctgcgc catcagatcc
     3361 ttggcggcaa gaaagccatc cagtttactt tgcagggctt cccaacctta ccagagggcg
     3421 ccccagctgg caattctttt gaagctcacg ctgccgcaag cactcagggc gcaagggctg
     3481 ctaaaggaag cggaacacgt agaaagccag tccgcagaaa cggtgctgac cccggatgaa
     3541 tgtcagctac tgggctatct ggacaaggga aaacgcaagc gcaaagagga tccattagaa
     3601 ttcgatttta ccacatttgt agaggtttta cttgcttaaa aaaacctccc acatctcccc
     3661 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     3721 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     3781 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg ctcgaagcgg
     3841 ccgccccaag gggttatgct agttattgct cagcggtggc agcagccaac tcagcttcct
     3901 ttcgggcttt gttagcagcc ggatcagctt gcatgcctgc aggtcgactc tagaattcta
     3961 ttaaccggaa atctccagag tagacagcca gcgcgcgatc tcgctgccga tcaggtccgg
     4021 gttgtcttct tgcagcagat tcagacccgg gccgatgtcc acagccttgc agttaggcag
     4081 gcttttggcc aggcgagcgg cttcggccgg tgggatcaga acgcctgggg tgccccagaa
     4141 cagcagcttc gggacagggg actggtgcag ccagtccatg tattcttcga ccagcgcgac
     4201 gatgttcgct ggctcaccgg cgattggcag ctcgtttggg aagcgccaca gtggctcgcg
     4261 gtcaacagga ttcaggaacg gctcgcggta atggtccatc tcgacttcag tcagcgggcg
     4321 gacgacaccc atcggcagcg taccctcgat aaaaacgttc tgatcgatga tcagcttgcg
     4381 gccgacgtcg gtggtgcgga aggcctggaa ggtctcgcgg gcaaattctg gccattcgtc
     4441 ccaggtcggg atagggcgga tgaactccat aaatgcaata cctttgacgc gctctggatt
     4501 gcgcttggcc cagtggaaac ccagagcgga gccccagtcg tgaatgacca ggacgacctc
     4561 ttccagaccc agggcttcga tgaaggcatc catgaagcgg acgtggtcgt cgaagaaata
     4621 acccaggtct ggtttgtcgg atttgcccat accgatcagg tctggagcaa tgcagcgatg
     4681 ggtcggtgca acatgcggga tgatgttgcg ccacacgtag gaggaggtcg ggttaccgtg
     4741 caggaacagc acaggggtgc catcgcgcgg accaacatcg acgtagtgca tgcgctcgcc
     4801 caggacttcc acataatggg ggtcgaatgg aaagccagta ccgatttcgg atccatcgtt
     4861 atcgctctga aagtacagat cctcagtggt tggctcgaga gaaacgcctc cgaagccgta
     4921 cagggtgcgc ccctggcgct tgagcgcgta caccacatcc atggctgtga cggtcttgcg
     4981 cttg
//
