LOCUS       sequence-470821-        9065 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express NanoLuc(R)-WRN 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 9065)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9065)
  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            "Unpublished"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: NanoLuc-WRN pFN31K Vector
FEATURES             Location/Qualifiers
     source          1..9065
                     /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(6008..6026)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    6063..6184
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6074..6093)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6128..6147
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        6429..6786
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6429..6620
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6452..6472)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6637..6772
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        6801..6848
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6880..7674
                     /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(6934..6953)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7544..7563
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7738..7786
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(8025..8613)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8105..8124)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    8729..9012
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
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 gagtgaaaaa aaattggaaa
     1621 caactgcaca gcagcggaaa tgtcctgaat ggatgaatgt gcagaataaa agatgtgctg
     1681 tagaagaaag aaaggcatgt gttcggaaga gtgtttttga agatgacctc cccttcttag
     1741 aattcactgg atccattgtg tatagttacg atgctagtga ttgctctttc ctgtcagaag
     1801 atattagcat gagtctatca gatggggatg tggtgggatt tgacatggag tggccaccat
     1861 tatacaatag agggaaactt ggcaaagttg cactaattca gttgtgtgtt tctgagagca
     1921 aatgttactt gttccacgtt tcttccatgt cagtttttcc ccagggatta aaaatgttgc
     1981 ttgaaaataa agcagttaaa aaggcaggtg taggaattga aggagatcag tggaaacttc
     2041 tacgtgactt tgatatcaaa ttgaagaatt ttgtggagtt gacagatgtt gccaataaaa
     2101 agctgaaatg cacagagacc tggagcctta acagtctggt taaacacctc ttaggtaaac
     2161 agctcctgaa agacaagtct atccgctgta gcaattggag taaatttcct ctcactgagg
     2221 accagaaact gtatgcagcc actgatgctt atgctggttt tattatttac cgaaatttag
     2281 agattttgga tgatactgtg caaaggtttg ctataaataa agaggaagaa atcctactta
     2341 gcgacatgaa caaacagttg acttcaatct ctgaggaagt gatggatctg gctaagcatc
     2401 ttcctcatgc tttcagtaaa ttggaaaacc cacggagggt ttctatctta ctaaaggata
     2461 tttcagaaaa tctatattca ctgaggagga tgataattgg gtctactaac attgagactg
     2521 aactgaggcc cagcaataat ttaaacttat tatcctttga agattcaact actgggggag
     2581 tacaacagaa acaaattaga gaacatgaag ttttaattca cgttgaagat gaaacatggg
     2641 acccaacact tgatcattta gctaaacatg atggagaaga tgtacttgga aataaagtgg
     2701 aacgaaaaga agatggattt gaagatggag tagaagacaa caaattgaaa gagaatatgg
     2761 aaagagcttg tttgatgtcg ttagatatta cagaacatga actccaaatt ttggaacagc
     2821 agtctcagga agaatatctt agtgatattg cttataaatc tactgagcat ttatctccca
     2881 atgataatga aaacgatacg tcctatgtaa ttgagagtga tgaagattta gaaatggaga
     2941 tgcttaagca tttatctccc aatgataatg aaaacgatac gtcctatgta attgagagtg
     3001 atgaagattt agaaatggag atgcttaagt ctttagaaaa cctcaatagt ggcacggtag
     3061 aaccaactca ttctaaatgc ttaaaaatgg aaagaaatct gggtcttcct actaaagaag
     3121 aagaagaaga tgatgaaaat gaagctaatg aaggggaaga agatgatgat aaggactttt
     3181 tgtggccagc acccaatgaa gagcaagtta cttgcctcaa gatgtacttt ggccattcca
     3241 gttttaaacc agttcagtgg aaagtgattc attcagtatt agaagaaaga agagataatg
     3301 ttgctgtcat ggcaactgga tatggaaaga gtttgtgctt ccagtatcca cctgtttatg
     3361 taggcaagat tggccttgtt atctctcccc ttatttctct gatggaagac caagtgctac
     3421 agcttaaaat gtccaacatc ccagcttgct tccttggatc agcacagtca gaaaatgttc
     3481 taacagatat taaattaggt aaataccgga ttgtatacgt aactccagaa tactgttcag
     3541 gtaacatggg cctgctccag caacttgagg ctgatattgg tatcacgctc attgctgtgg
     3601 atgaggctca ctgtatttct gagtgggggc atgattttag ggattcattc aggaagttgg
     3661 gctccctaaa gacagcactg ccaatggttc caatcgttgc acttactgct actgcaagtt
     3721 cttcaatccg ggaagacatt gtacgttgct taaatctgag aaatcctcag atcacctgta
     3781 ctggttttga tcgaccaaac ctgtatttag aagttaggcg aaaaacaggg aatatccttc
     3841 aggatctgca gccatttctt gtcaaaacaa gttcccactg ggaatttgaa ggtccaacaa
     3901 tcatctactg tccttctaga aaaatgacac aacaagttac aggtgaactt aggaaactga
     3961 atctatcctg tggaacatac catgcgggca tgagttttag cacaaggaaa gacattcatc
     4021 ataggtttgt aagagatgaa attcagtgtg tcatagctac catagctttt ggaatgggca
     4081 ttaataaagc tgacattcgc caagtcattc attacggtgc tcctaaggac atggaatcat
     4141 attatcagga gattggtaga gctggtcgtg atggacttca aagttcttgt cacgtcctct
     4201 gggctcctgc agacattaac ttaaataggc accttcttac tgagatacgt aatgagaagt
     4261 ttcgattata caaattaaag atgatggcaa agatggaaaa atatcttcat tctagcagat
     4321 gtaggagaca aatcatcttg tctcattttg aggacaaaca agtacaaaaa gcctccttgg
     4381 gaattatggg aactgaaaaa tgctgtgata attgcaggtc cagattggat cattgctatt
     4441 ccatggatga ctcagaggat acatcctggg actttggtcc acaagcattt aagcttttgt
     4501 ctgctgtgga catcttaggc gaaaaatttg gaattgggct tccaatttta tttctccgag
     4561 gatctaattc tcagcgtctt gccgatcaat atcgcaggca cagtttattt ggcactggca
     4621 aggatcaaac agagagttgg tggaaggctt tttcccgtca gctgatcact gagggattct
     4681 tggtagaagt ttctcggtat aacaaattta tgaagatttg cgcccttacg aaaaagggta
     4741 gaaattggct tcataaagct aatacagaat ctcagagcct catccttcaa gctaatgaag
     4801 aattgtgtcc aaagaagttg cttctgccta gttcgaaaac tgtatcttcg ggcaccaaag
     4861 agcattgtta taatcaagta ccagttgaat taagtacaga gaagaagtct aacttggaga
     4921 agttatattc ttataaacca tgtgataaga tttcttctgg gagtaacatt tctaaaaaaa
     4981 gtatcatggt acagtcacca gaaaaagctt acagttcctc acagcctgtt atttcggcac
     5041 aagagcagga gactcagatt gtgttatatg gcaaattggt agaagctagg cagaaacatg
     5101 ccaataaaat ggatgttccc ccagctattc tggcaacaaa caagatactg gtggatatgg
     5161 ccaaaatgag accaactacg gttgaaaacg taaaaaggat tgatggtgtt tctgaaggca
     5221 aagctgccat gttggcccct ctgttggaag tcatcaaaca tttctgccaa acaaatagtg
     5281 ttcagacaga cctcttttca agtacaaaac ctcaagaaga acagaagacg agtctggtag
     5341 caaaaaataa aatatgcaca ctttcacagt ctatggccat cacatactct ttattccaag
     5401 aaaagaagat gcctttgaag agcatagctg agagcaggat tctgcctctc atgacaattg
     5461 gcatgcactt atcccaagcg gtgaaagctg gctgccccct tgatttggag cgagcaggcc
     5521 tgactccaga ggttcagaag attattgctg atgttatccg aaaccctccc gtcaactcag
     5581 atatgagtaa aattagccta atcagaatgt tagttcctga aaacattgac acgtacctta
     5641 tccacatggc aattgagatc cttaaacatg gtcctgacag cggacttcaa ccttcatgtg
     5701 atgtcaacaa aaggagatgt tttcccggtt ctgaagagat ctgttcaagt tctaagagaa
     5761 gcaaggaaga agtaggcatc aatactgaga cttcatctgc agagagaaag agacgattac
     5821 ctgtgtggtt tgccaaagga agtgatacca gcaagaaatt aatggacaaa acgaaaaggg
     5881 gaggtctttt tagtgtttaa acgaattcgg gctcggtacc cggggatcct ctagagtcga
     5941 cctgcaggca tgcaagctga tccggctgct aacaaagccc gaaaggaagc tgagttggct
     6001 gctgccaccg ctgagcaata actagcataa ccccttgggg cggccgcttc gagcagacat
     6061 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     6121 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     6181 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt
     6241 ttttttaagc aagtaaaacc tctacaaatg tggtaaaatc gaattttaac aaaatattaa
     6301 cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     6361 catacgcgga tctgcgcagc accatggcct gaaataacct ctgaaagagg aacttggtta
     6421 ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     6481 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     6541 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     6601 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     6661 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     6721 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     6781 tgcaaaaagc ttaattaact gttgacaatt aatcatcggc atagtatatc ggcatagtat
     6841 aatacgacaa ggtgaggaac taaacccagg aggcagatca tgattgaaca agatggattg
     6901 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     6961 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     7021 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     7081 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     7141 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     7201 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     7261 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     7321 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     7381 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     7441 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     7501 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     7561 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     7621 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     7681 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga tggccgcaat
     7741 aaaatatctt tattttcatt acatctgtgt gttggttttt tgtgtgaatc gatagcgata
     7801 aggatcctct ttgcgcttgc gttttccctt gtccagatag cccagtagct gacattcatc
     7861 cggggtcagc accgtttctg cggactggct ttctacccgg tatcagctca ctcaaaggcg
     7921 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     7981 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     8041 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     8101 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     8161 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     8221 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     8281 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     8341 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     8401 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     8461 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     8521 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     8581 cagcagatta cgcgcagaaa aaaaggattt caagaagatc ctttgatctt ttctacgggg
     8641 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     8701 aggatcttca cctagatcct tttatagtcc ggaaatacag gaacgcacgc tggatggccc
     8761 ttcgctggga tggtgaaacc atgaaaaatg gcagcttcag tggattaagt gggggtaatg
     8821 tggcctgtac cctctggttg cataggtatt catacggtta aaatttatca ggcgcgattg
     8881 cggcagtttt tcgggtggtt tgttgccatt tttacctgtc tgctgccgtg atcgcgctga
     8941 acgcgtttta gcggtgcgta caattaaggg attatggtaa atccacttac tgtctgccct
     9001 cgtagccatc gagataaacc gcagtactcc ggccacgatg cgtccggcgt agaggatcga
     9061 gatct
//
