LOCUS       sequence-501959-        8903 bp    DNA     circular SYN 04-MAR-2026
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 8903)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8903)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 4, 2026 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            "Unpublished"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..8903
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(3521..3540)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3522..3540)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3582..3601)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          3584..3716
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(3852..4055)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3881..3901)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4056..4435
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(4880..4897)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5014..5559)
                     /direction=LEFT
                     /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     complement(5094..5113)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5716..5764
                     /note="synthetic polyadenylation signal"
     CDS             complement(5828..6622)
                     /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(5939..5958)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6549..6568
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(6654..6701)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(6716..7073)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6730..6865
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6882..7073
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     7030..7050
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    7318..7439
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7355..7374)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7409..7428
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7476..7494
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(7582..8094)
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
ORIGIN
        1 tgttcgcatg cttctgccgc gcttcaacaa gtttgccata aagaactatc tgcgtttcct
       61 gttcctgggc agatatcacg ggctgtgaag agctataagc cttttcgggt gattggacca
      121 tgatagattt cttagatata ttgctgccag aactaatttt gtcacaaggt ttgtagctat
      181 agagcttctc caagttagat ttcttttccg tggacagctc tactggaacc tggttatagc
      241 aatgttcctt cgtaccgctg gacactgttt tactgctcgg caacaaaagc ttcttagggc
      301 aaagctcttc gttagcttgc aagatcaagg actgtgactc ggtgttcgct ttgtgaagcc
      361 aattgcgccc tttcttcgtc agcgcgcaga ttttcataaa cttgttatac cgactcactt
      421 caaccagaaa gccttcggtt atcagctggc gactaaaggc cttccaccag gactctgttt
      481 gatcctttcc agtcccgaac aggctgtgcc ggcgatactg atcagcgagt cgttgactat
      541 tggaacctcg caaaaagagt atcgggagtc cgatgccaaa cttttcaccc aatatgtcaa
      601 ccgcgctcag cagcttaaaa gcttgcggac caaaatccca ggaggtatcc tcgctgtcat
      661 ccatactgta gcaatgatcc aagcgtgacc tgcagttgtc gcaacacttc tcggtcccca
      721 tgattccaag agatgctttc tggacctgct tgtcttcgaa atgactaaga ataatctggc
      781 ggcggcatct tgaagagtgg agatatttct ccatcttcgc catcatttta agtttataaa
      841 ggcggaactt ttcgttgcgt atttcggtga gcaagtgccg gttaaggttt atatcagcag
      901 gcgcccacaa gacgtggcaa gagctttgaa gtccgtcgcg cccggcgcga ccgatctctt
      961 gatagtatga ctccatgtct ttaggcgcac catagtgtat gacttgtcgt atgtccgcct
     1021 tgttaatgcc catcccgaat gcgatcgttg cgatgacaca ctgtatttca tcccggacga
     1081 agcggtgatg tatatctttt cttgtgctaa aggacatccc cgcgtggtac gtcccacaac
     1141 tgaggttgag tttgcggagt tcgcccgtaa cttgctgtgt catcttccga ctcgggcagt
     1201 atatgatagt aggtccttca aattcccaat ggctactcgt ttttaccaaa aatggttgca
     1261 ggtcctggag tatgttcccg gtcttccgcc gcacttccaa atagaggttg ggtctgtcga
     1321 atcctgtgca ggtaatctgc gggtttcgca agttcagaca tcgcacgatg tcttccctaa
     1381 tggagctgct tgccgttgcg gtcaaggcaa cgatcggaac catgggcaaa gccgttttca
     1441 aacttccgag cttcctaaaa gaatctctga aatcgtgacc ccattcggat atacaatggg
     1501 cttcatctac agctatgagg gtaattccga tatcagcctc cagctgctgg agcagtccca
     1561 tattaccaga acagtactcg ggcgttacat atacgattct atattttccg agtttaatgt
     1621 ccgtcaggac attctcactt tgcgctgaac caagaaaaca ggccggaatg ttggacatct
     1681 taagttgcaa tacttggtcc tccatcagac ttatgagcgg gcttataacg agtccgatct
     1741 tccctacata aacaggtggg tactgaaaac acagggactt gccataccca gttgccatga
     1801 cggcgacatt gtccctcctc tcctccagta cagagtgaat gactttccat tggacaggtt
     1861 tgaaagagga gtgtccgaaa tacatcttca gacaagttac ttgttcctcg ttaggcgccg
     1921 gccaaagaaa gtccttatca tcatcctctt caccctcgtt cgcttcattt tcgtcatcct
     1981 cctcctcttc tttggtagga aggccgaggt tcctttccat tttaaggcac ttactgtgtg
     2041 taggttcgac ggtacctgag ttcaaattct cgagtgactt gagcatttcc atttccagat
     2101 cctcatcaga ttctattaca taagaagtgt cgttttcgtt gtcatttggt gacagatgct
     2161 taagcatttc catttcgagg tcctcgtcag attcaattac ataggaggtg tcattctcat
     2221 tgtcgtttgg agacaggtgt tctgtacttt tgtacgcgat gtcgctgaga tattcttcct
     2281 gtgactgttg ctcaaggatc tgcagctcgt gttcagttat gtcgaggctc ataagacatg
     2341 cccgctccat attttctttg agtttattat cctctacccc atcttcgaag ccgtcctctt
     2401 tgcgctcgac cttattaccg agaacatctt ctccgtcatg cttggccaaa tgatcgagtg
     2461 tcggatccca cgtttcgtcc tccacatgga tcaaaacctc atgttcccga atctgtttct
     2521 gttggacccc acccgttgtg ctatcctcaa aactaagcaa attcaagtta ttactaggcc
     2581 tcagctcggt ttcaatgttc gtgctgccga tgatcatccg tctgagtgag tagagattct
     2641 cagaaatgtc tttaagcagt atggatactc ggcgtgggtt ctcgagctta ctaaaggcat
     2701 gtggcaagtg ctttgcaaga tccatcactt cctcagatat tgatgtgagc tgcttattca
     2761 tatcagacaa caaaatttcc tcctctttat tgattgcaaa ccgctgtacc gtatcgtcca
     2821 atatctccaa attccggtat ataataaaac ctgcgtaagc gtcggttgca gcgtacagtt
     2881 tctgatcttc cgtcagcgga aacttggacc agttgctgca tcgtatactc ttgtctttca
     2941 acaactgctt tccgagcagg tgttttacga gtgagttaag ggaccacgtc tcagtacact
     3001 taagcttttt attagccacg tcagtaagct ctacgaagtt cttaagttta atgtcaaaat
     3061 ctctcaacaa cttccattgg tccccttcta tgcctactcc cgcttttttc accgccttgt
     3121 tctcgagcag cattttgagc ccctgcggga aaacggacat gctggaaaca tggaacaaat
     3181 agcatttgct ctcactgacg caaagctgta taagggcgac cttgccgagc ttccccctgt
     3241 tgtacaaggg cggccactcc atatcaaagc cgaccacgtc accgtcgctc aaggacatac
     3301 taatatcttc actcaggaag gagcaatcgc ttgcgtcgta tgaataaaca attgatccgg
     3361 tgaactcaag gaacggaaga tcatcttcaa agactgattt ccgcacacaa gctttgcgct
     3421 cttctaccgc acatctcttg ttttgaacgt tcatccactc ggggcatttg cgctgttgtg
     3481 cagtggtctc aagttttttt tcggacatgg cgatcgctag ccctatagtg agtcgtatta
     3541 atactctagc cttaagagct gtaattgaac tgggagtgga cacctgtgga gagaaaggca
     3601 aagtggatgt cagtaagacc aataggtgcc tatcagaaac gcaagagtct tctctgtctc
     3661 gacaagccca gtttctattg gtctccttaa acctgtcttg taaccttgat acttacctgc
     3721 caagtgcctc acgaccaact tctgcagctt aagttcgaga ctgttgtgtc agaagcactg
     3781 actgcgttag caatttaact gtgataaact accgcaataa agcttctagt gatctgacgg
     3841 ttcactaaac cagctctgct tatatagacc tcccaccgta cacgcctacc gcccatttgc
     3901 gtcaacgggg cggggttatt acgacatttt ggaaagtccc gttgattttg gtgccaaaac
     3961 aaactcccat tgacgtcaat ggggtggaga cttggaaatc cccgtgagtc aaaccgctat
     4021 ccacgcccat tggtgtactg ccaaaaccgc atcaccatgg taatagcgat gactaatacg
     4081 tagatgtact gccaagtagg aaagtcccgt aaggtcatgt actgggcata atgccaggcg
     4141 ggccatttac cgtcattgac gtcaataggg ggcggacttg gcatatgata cacttgatgt
     4201 actgccaagt gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct
     4261 attggcgtta ctatgggaac atacgtcatt attgacgtca atgggcgggg gtcgttgggc
     4321 ggtcagccag gcgggccatt taccgtaagt tatgtaacgc ggaactccat atatgggcta
     4381 tgaactaatg accccgtaat tgattactat taataactag tcaataatca atgccaacat
     4441 ggcggtcata ttggacatga gccaatataa atgtacatat tatgatatag atacaacgta
     4501 tgcaatggcc aatagccaat attgatttat gctatataac caatgaataa tatggctaat
     4561 ggccaatatt gaagatctcg atcctctacg ccggacgcat cgtggccgga gtactgcggt
     4621 ttatctcgat ggctacgagg gcagacagta agtggattta ccataatccc ttaattgtac
     4681 gcaccgctaa aacgcgtaat ctcatgacgg ggtggtgcgt aacggcaaaa gcaccgccgg
     4741 acatcagcgc tagcggagtg tatactggct tactatgttg gcactgatga gggtgtcagt
     4801 gaagtgcttc atgtggcagg agaaaaaagg ctgcaccggt gcgtcagcag aatatgtgat
     4861 acaggatata ttccgcttcc tcgctcactg actcgctacg ctcggtcgtt cgactgcggc
     4921 gagcggaaat ggcttacgaa cggggcggag atttcctgga agatgccagg aagatactta
     4981 acagggaagt gagagggccg cggcaaagcc gtttttccat aggctccgcc cccctgacaa
     5041 gcatcacgaa atctgacgct caaatcagtg gtggcgaaac ccgacaggac tataaagata
     5101 ccaggcgttt ccccctggcg gctccctcgt gcgctctcct gttcctgcct ttcggtttac
     5161 cggtgtcatt ccgctgttat ggccgcgttt gtctcattcc acgcctgaca ctcagttccg
     5221 ggtaggcagt tcgctccaag ctggactgta tgcacgaacc ccccgttcag tccgaccgct
     5281 gcgccttatc cggtaactat cgtcttgagt ccaacccgga aagacatgca aaagcaccac
     5341 tggcagcagc cactggtaat tgatttagag gagttagtct tgaagtcatg cgccggttaa
     5401 ggctaaactg aaaggacaag ttttggtgac tgcgctcctc caagccagtt acctcggttc
     5461 aaagagttgg tagctcagag aaccttcgaa aaaccgccct gcaaggcggt tttttcgttt
     5521 tcagagcaag agattacgcg cagaccaaaa cgatctcaag aagatcatct tattaatcag
     5581 ataaaatatt tctagatttc agtgcaattt atctcttcaa atgtagcacc tgaagtcagc
     5641 cccatacgat ataagttgta attctcatgt ttgacagctt atcatcgcat gagcggatcc
     5701 ttatcgctat cgattcacac aaaaaaccaa cacacagatg taatgaaaat aaagatattt
     5761 tattgcggcc atcgtgatgg caggttgggc gtcgcttggt cggtcatttc gaaccccaga
     5821 gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag
     5881 cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc tcttcagcaa
     5941 tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc cggccacagt
     6001 cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag gcatcgccat
     6061 gggtcacgac gagatcctcg ccgtcgggca tgcgcgcctt gagcctggcg aacagttcgg
     6121 ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga ccggcttcca
     6181 tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg
     6241 gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc tcggcaggag
     6301 caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc cagtcccttc
     6361 ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg gccagccacg
     6421 atagccgcgc tgcctcgtcc tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa
     6481 aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag cagccgattg
     6541 tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga gaacctgcgt
     6601 gcaatccatc ttgttcaatc atgatctgcc tcctgggttt agttcctcac cttgtcgtat
     6661 tatactatgc cgatatacta tgccgatgat taattgtcaa cagttaatta agctttttgc
     6721 aaaagcctag gcctccaaaa aagcctcctc actacttctg gaatagctca gaggccgagg
     6781 cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg
     6841 aactgggcgg agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact
     6901 aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc
     6961 tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg
     7021 actttccaca ccctaactga cacacattcc acagctggtt ctttccgcct cagaaggtac
     7081 ctaaccaagt tcctctttca gaggttattt caggccatgg tgctgcgcag atccgcgtat
     7141 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcagga aattgtaagc
     7201 gttaatattt tgttaaaatt cgattttacc acatttgtag aggttttact tgcttaaaaa
     7261 aacctcccac atctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     7321 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     7381 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     7441 catgtctgct cgaagcggcc gccccaaggg gttatgctag ttattgctca gcggtggcag
     7501 cagccaactc agcttccttt cgggctttgt tagcagccgg atcagcttgc atgcctgcag
     7561 gtcgactcta gagtcgcggc cttacgccag aatgcgttcg cacagccgcc agccggtcac
     7621 tccgttgatg gttactcgga acagcaggga gccgtcgggg ttgatcaggc gctcgtcgat
     7681 aattttgttg ccgttccaca gggtccctgt tacagtgatc tttttgccgt cgaacacggc
     7741 gatgccttca tacggccgtc cgaaatagtc gatcatgttc ggcgtaaccc cgtcgattac
     7801 cagtgtgcca tagtgcagga tcaccttaaa gtgatgatca tccacagggt acaccacctt
     7861 aaaaattttt tcgatctggc ccatttggtc gccgctcaga ccttcatacg ggatgatgac
     7921 atggatgtcg atcttcagcc cattttcacc gctcaggaca atcctttgga tcggagttac
     7981 ggacaccccg agattctgaa acaaactgga cacacctccc tgttcaagga cttggtccag
     8041 gttgtagccg gctgtctgtc gccagtcccc aacgaaatct tcgagtgtga agacgccgct
     8101 cgagccgaga gaaacagaga acaggccacc tcgcttggtt ttgtccatga gtttcttgct
     8161 tgtatcgctt cccttcgcga accatactgg caacctcctc tttctttctg ctgaagaggt
     8221 ctctgtattt ataccaacct cctctttaga ccgtttactg ctactacaga tttcttcact
     8281 ccctgggaaa catcggcgtt tgttcacgtc acaactaggc tggaggccgc tgtcgggtcc
     8341 atgtttaagg atctcaatcg ccatatggat aaggtaagta tcaatattct ccgggaccag
     8401 catgcggatc aatgagatct tactcatgtc tgagttgact ggtgggtttc ttattacgtc
     8461 cgctatgatc ttttggactt ctggtgtcaa ccccgccctc tcaaggtcaa gcggacatcc
     8521 cgctttcact gcctgtgaaa gatgcatacc gatggtcatc aatggaagga ttcttgattc
     8581 agcgattgac ttaaggggca tcttcttctc ttggaaaagt gaatatgtta tagccataga
     8641 ttgggacaag gtgcaaatct tgtttttggc aacaagagac gtcttttgct cctcttgtgg
     8701 cttagtactg ctaaagagat ccgtttgcac actattcgtc tggcaaaaat gtttaattac
     8761 ttcaagcaac ggagcaagca ttgccgcttt tccttcgctc accccgtcga tccgtttaac
     8821 gttttctacg gtggtggggc gcatcttcgc catatctaca agtattttgt tcgtcgccaa
     8881 gatggccggt gggacatcca ttt
//
