LOCUS       sequence-476794-        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  JOURNAL   Unpublished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            "Unpublished"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(252..270)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    307..428
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(318..337)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     372..391
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             827..1621
                     /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(881..900)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1491..1510
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(1793..2381)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1873..1892)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2497..2780
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     enhancer        2971..3350
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3351..3554
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3505..3525
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          3690..3822
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3805..3824
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     3866..3885
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3866..3884
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4228..4248
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             join(4264..4984,1..164)
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
ORIGIN
        1 caggcgttct gatcccaccg gccgaagccg ctcgcctggc caaaagcctg cctaactgca
       61 aggctgtgga catcggcccg ggtctgaatc tgctgcaaga agacaacccg gacctgatcg
      121 gcagcgagat cgcgcgctgg ctgtctactc tggagatttc cggttaatag aattctagag
      181 tcgacctgca ggcatgcaag ctgatccggc tgctaacaaa gcccgaaagg aagctgagtt
      241 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcggccg cttcgagcag
      301 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
      361 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
      421 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg
      481 aggttttttt aagcaagtaa aacctctaca aatgtggtaa aatcgaattc taatggatcc
      541 tctttgcgct tgcgttttcc cttgtccaga tagcccagta gctgacattc atccggggtc
      601 agcaccgttt ctgcggactg gctttctacg tgttccgctt cctttagcag cccttgcgcc
      661 ctgagtgctt gcggcagcgt gagcttcaaa agaattgcca gctggggcgc cctctggtaa
      721 ggttgggaag ccctgcaaag taaactggat ggctttcttg ccgccaagga tctgatggcg
      781 caggggatca agatctgatc aagagacagg atgacggtcg tttcgcatgc ttgaacaaga
      841 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
      901 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
      961 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     1021 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     1081 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     1141 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     1201 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     1261 cactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     1321 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgtatg ccggatggtg aggatctcgt
     1381 cgtgactcat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     1441 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     1501 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     1561 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     1621 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaaccggta tcagctcact
     1681 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     1741 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     1801 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     1861 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     1921 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     1981 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     2041 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     2101 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     2161 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     2221 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     2281 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     2341 tttgcaagca gcagattacg cgcagaaaaa aaggatttca agaagatcct ttgatctttt
     2401 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     2461 tatcaaaaag gatcttcacc tagatccttt tatagtccgg aaatacagga acgcacgctg
     2521 gatggccctt cgctgggatg gtgaaaccat gaaaaatggc agcttcagtg gattaagtgg
     2581 gggtaatgtg gcctgtaccc tctggttgca taggtattca tacggttaaa atttatcagg
     2641 cgcgattgcg gcagtttttc gggtggtttg ttgccatttt tacctgtctg ctgccgtgat
     2701 cgcgctgaac gcgttttagc ggtgcgtaca attaagggat tatggtaaat ccacttactg
     2761 tctgccctcg tagccatcga gataaaccgc agtactccgg ccacgatgcg tccggcgtag
     2821 aggatcgaga tcttcaatat tggccattag ccatattatt cattggttat atagcataaa
     2881 tcaatattgg ctattggcca ttgcatacgt tgtatctata tcataatatg tacatttata
     2941 ttggctcatg tccaatatga ccgccatgtt ggcattgatt attgactagt tattaatagt
     3001 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     3061 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     3121 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     3181 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt ccgcccccta
     3241 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttacggg
     3301 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
     3361 tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     3421 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     3481 gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     3541 atataagcag agctggttta gtgaaccgtc agatcactag aagctttatt gcggtagttt
     3601 atcacagtta aattgctaac gcagtcagtg cttctgacac aacagtctcg aacttaagct
     3661 gcagaagttg gtcgtgaggc actgggcagg taagtatcaa ggttacaaga caggtttaag
     3721 gagaccaata gaaactgggc ttgtcgagac agagaagact cttgcgtttc tgataggcac
     3781 ctattggtct tactgacatc cactttgcct ttctctccac aggtgtccac tcccagttca
     3841 attacagctc ttaaggctag agtattaata cgactcacta tagggctagc gatcgccatg
     3901 tccggcagag gaaagggcgg aaaaggctta ggcaaagggg gcgctaagcg ccaccgcaag
     3961 gtcttgagag acaacattca gggcatcacc aagcctgcca ttcggcgtct agctcggcgt
     4021 ggcggcgtta agcggatctc tggcctcatt tacgaggaga cccgcggtgt gctgaaggtg
     4081 ttcctggaga atgtgattcg ggacgcagtc acctacaccg agcacgccaa gcgcaagacc
     4141 gtcacagcca tggatgtggt gtacgcgctc aagcgccagg ggcgcaccct gtacggcttc
     4201 ggaggcgttt ctctcgagcc aaccactgag gatctgtact ttcagagcga taacgatgga
     4261 tccgaaatcg gtactggctt tccattcgac ccccattatg tggaagtcct gggcgagcgc
     4321 atgcactacg tcgatgttgg tccgcgcgat ggcacccctg tgctgttcct gcacggtaac
     4381 ccgacctcct cctacgtgtg gcgcaacatc atcccgcatg ttgcaccgac ccatcgctgc
     4441 attgctccag acctgatcgg tatgggcaaa tccgacaaac cagacctggg ttatttcttc
     4501 gacgaccacg tccgcttcat ggatgccttc atcgaagccc tgggtctgga agaggtcgtc
     4561 ctggtcattc acgactgggg ctccgctctg ggtttccact gggccaagcg caatccagag
     4621 cgcgtcaaag gtattgcatt tatggagttc atccgcccta tcccgacctg ggacgaatgg
     4681 ccagaatttg cccgcgagac cttccaggcc ttccgcacca ccgacgtcgg ccgcaagctg
     4741 atcatcgatc agaacgtttt tatcgagggt acgctgccga tgggtgtcgt ccgcccgctg
     4801 actgaagtcg agatggacca ttaccgcgag ccgttcctga atcctgttga ccgcgagcca
     4861 ctgtggcgct tcccaaacga gctgccaatc gccggtgagc cagcgaacat cgtcgcgctg
     4921 gtcgaagaat acatggactg gctgcaccag tcccctgtcc cgaagctgct gttctggggc
     4981 accc
//
