LOCUS       sequence-470995-        9039 bp    DNA     circular SYN 06-MAY-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 9039)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 9039)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 6, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: WRN-NanoLuc pFC32K Vector
FEATURES             Location/Qualifiers
     source          1..9039
                     /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             5382..5894
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(5982..6000)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    6037..6158
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6048..6067)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6102..6121
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        6403..6760
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6403..6594
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6426..6446)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6611..6746
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        6775..6822
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6854..7648
                     /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(6908..6927)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7518..7537
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7712..7760
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(7999..8587)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8079..8098)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    8703..8986
                     /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 ggctagcgat cgccatgagt gaaaaaaaat
     1081 tggaaacaac tgcacagcag cggaaatgtc ctgaatggat gaatgtgcag aataaaagat
     1141 gtgctgtaga agaaagaaag gcatgtgttc ggaagagtgt ttttgaagat gacctcccct
     1201 tcttagaatt cactggatcc attgtgtata gttacgatgc tagtgattgc tctttcctgt
     1261 cagaagatat tagcatgagt ctatcagatg gggatgtggt gggatttgac atggagtggc
     1321 caccattata caatagaggg aaacttggca aagttgcact aattcagttg tgtgtttctg
     1381 agagcaaatg ttacttgttc cacgtttctt ccatgtcagt ttttccccag ggattaaaaa
     1441 tgttgcttga aaataaagca gttaaaaagg caggtgtagg aattgaagga gatcagtgga
     1501 aacttctacg tgactttgat atcaaattga agaattttgt ggagttgaca gatgttgcca
     1561 ataaaaagct gaaatgcaca gagacctgga gccttaacag tctggttaaa cacctcttag
     1621 gtaaacagct cctgaaagac aagtctatcc gctgtagcaa ttggagtaaa tttcctctca
     1681 ctgaggacca gaaactgtat gcagccactg atgcttatgc tggttttatt atttaccgaa
     1741 atttagagat tttggatgat actgtgcaaa ggtttgctat aaataaagag gaagaaatcc
     1801 tacttagcga catgaacaaa cagttgactt caatctctga ggaagtgatg gatctggcta
     1861 agcatcttcc tcatgctttc agtaaattgg aaaacccacg gagggtttct atcttactaa
     1921 aggatatttc agaaaatcta tattcactga ggaggatgat aattgggtct actaacattg
     1981 agactgaact gaggcccagc aataatttaa acttattatc ctttgaagat tcaactactg
     2041 ggggagtaca acagaaacaa attagagaac atgaagtttt aattcacgtt gaagatgaaa
     2101 catgggaccc aacacttgat catttagcta aacatgatgg agaagatgta cttggaaata
     2161 aagtggaacg aaaagaagat ggatttgaag atggagtaga agacaacaaa ttgaaagaga
     2221 atatggaaag agcttgtttg atgtcgttag atattacaga acatgaactc caaattttgg
     2281 aacagcagtc tcaggaagaa tatcttagtg atattgctta taaatctact gagcatttat
     2341 ctcccaatga taatgaaaac gatacgtcct atgtaattga gagtgatgaa gatttagaaa
     2401 tggagatgct taagcattta tctcccaatg ataatgaaaa cgatacgtcc tatgtaattg
     2461 agagtgatga agatttagaa atggagatgc ttaagtcttt agaaaacctc aatagtggca
     2521 cggtagaacc aactcattct aaatgcttaa aaatggaaag aaatctgggt cttcctacta
     2581 aagaagaaga agaagatgat gaaaatgaag ctaatgaagg ggaagaagat gatgataagg
     2641 actttttgtg gccagcaccc aatgaagagc aagttacttg cctcaagatg tactttggcc
     2701 attccagttt taaaccagtt cagtggaaag tgattcattc agtattagaa gaaagaagag
     2761 ataatgttgc tgtcatggca actggatatg gaaagagttt gtgcttccag tatccacctg
     2821 tttatgtagg caagattggc cttgttatct ctccccttat ttctctgatg gaagaccaag
     2881 tgctacagct taaaatgtcc aacatcccag cttgcttcct tggatcagca cagtcagaaa
     2941 atgttctaac agatattaaa ttaggtaaat accggattgt atacgtaact ccagaatact
     3001 gttcaggtaa catgggcctg ctccagcaac ttgaggctga tattggtatc acgctcattg
     3061 ctgtggatga ggctcactgt atttctgagt gggggcatga ttttagggat tcattcagga
     3121 agttgggctc cctaaagaca gcactgccaa tggttccaat cgttgcactt actgctactg
     3181 caagttcttc aatccgggaa gacattgtac gttgcttaaa tctgagaaat cctcagatca
     3241 cctgtactgg ttttgatcga ccaaacctgt atttagaagt taggcgaaaa acagggaata
     3301 tccttcagga tctgcagcca tttcttgtca aaacaagttc ccactgggaa tttgaaggtc
     3361 caacaatcat ctactgtcct tctagaaaaa tgacacaaca agttacaggt gaacttagga
     3421 aactgaatct atcctgtgga acataccatg cgggcatgag ttttagcaca aggaaagaca
     3481 ttcatcatag gtttgtaaga gatgaaattc agtgtgtcat agctaccata gcttttggaa
     3541 tgggcattaa taaagctgac attcgccaag tcattcatta cggtgctcct aaggacatgg
     3601 aatcatatta tcaggagatt ggtagagctg gtcgtgatgg acttcaaagt tcttgtcacg
     3661 tcctctgggc tcctgcagac attaacttaa ataggcacct tcttactgag atacgtaatg
     3721 agaagtttcg attatacaaa ttaaagatga tggcaaagat ggaaaaatat cttcattcta
     3781 gcagatgtag gagacaaatc atcttgtctc attttgagga caaacaagta caaaaagcct
     3841 ccttgggaat tatgggaact gaaaaatgct gtgataattg caggtccaga ttggatcatt
     3901 gctattccat ggatgactca gaggatacat cctgggactt tggtccacaa gcatttaagc
     3961 ttttgtctgc tgtggacatc ttaggcgaaa aatttggaat tgggcttcca attttatttc
     4021 tccgaggatc taattctcag cgtcttgccg atcaatatcg caggcacagt ttatttggca
     4081 ctggcaagga tcaaacagag agttggtgga aggctttttc ccgtcagctg atcactgagg
     4141 gattcttggt agaagtttct cggtataaca aatttatgaa gatttgcgcc cttacgaaaa
     4201 agggtagaaa ttggcttcat aaagctaata cagaatctca gagcctcatc cttcaagcta
     4261 atgaagaatt gtgtccaaag aagttgcttc tgcctagttc gaaaactgta tcttcgggca
     4321 ccaaagagca ttgttataat caagtaccag ttgaattaag tacagagaag aagtctaact
     4381 tggagaagtt atattcttat aaaccatgtg ataagatttc ttctgggagt aacatttcta
     4441 aaaaaagtat catggtacag tcaccagaaa aagcttacag ttcctcacag cctgttattt
     4501 cggcacaaga gcaggagact cagattgtgt tatatggcaa attggtagaa gctaggcaga
     4561 aacatgccaa taaaatggat gttcccccag ctattctggc aacaaacaag atactggtgg
     4621 atatggccaa aatgagacca actacggttg aaaacgtaaa aaggattgat ggtgtttctg
     4681 aaggcaaagc tgccatgttg gcccctctgt tggaagtcat caaacatttc tgccaaacaa
     4741 atagtgttca gacagacctc ttttcaagta caaaacctca agaagaacag aagacgagtc
     4801 tggtagcaaa aaataaaata tgcacacttt cacagtctat ggccatcaca tactctttat
     4861 tccaagaaaa gaagatgcct ttgaagagca tagctgagag caggattctg cctctcatga
     4921 caattggcat gcacttatcc caagcggtga aagctggctg cccccttgat ttggagcgag
     4981 caggcctgac tccagaggtt cagaagatta ttgctgatgt tatccgaaac cctcccgtca
     5041 actcagatat gagtaaaatt agcctaatca gaatgttagt tcctgaaaac attgacacgt
     5101 accttatcca catggcaatt gagatcctta aacatggtcc tgacagcgga cttcaacctt
     5161 catgtgatgt caacaaaagg agatgttttc ccggttctga agagatctgt tcaagttcta
     5221 agagaagcaa ggaagaagta ggcatcaata ctgagacttc atctgcagag agaaagagac
     5281 gattacctgt gtggtttgcc aaaggaagtg ataccagcaa gaaattaatg gacaaaacga
     5341 aaaggggagg tctttttagt gtttctctcg gctcgagcgg cgtcttcaca ctcgaagatt
     5401 tcgttgggga ctggcgacag acagccggct acaacctgga ccaagtcctt gaacagggag
     5461 gtgtgtccag tttgtttcag aatctcgggg tgtccgtaac tccgatccaa aggattgtcc
     5521 tgagcggtga aaatgggctg aagatcgaca tccatgtcat catcccgtat gaaggtctga
     5581 gcggcgacca aatgggccag atcgaaaaaa tttttaaggt ggtgtaccct gtggatgatc
     5641 atcactttaa ggtgatcctg cactatggca cactggtaat cgacggggtt acgccgaaca
     5701 tgatcgacta tttcggacgg ccgtatgaag gcatcgccgt gttcgacggc aaaaagatca
     5761 ctgtaacagg gaccctgtgg aacggcaaca aaattatcga cgagcgcctg atcaaccccg
     5821 acggctccct gctgttccga gtaaccatca acggagtgac cggctggcgg ctgtgcgaac
     5881 gcattctggc gtaaggccgc gactctagag tcgacctgca ggcatgcaag ctgatccggc
     5941 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
     6001 ataacccctt ggggcggccg cttcgagcag acatgataag atacattgat gagtttggac
     6061 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     6121 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     6181 ttatgtttca ggttcagggg gagatgtggg aggttttttt aagcaagtaa aacctctaca
     6241 aatgtggtaa aatcgaattt taacaaaata ttaacgctta caatttcctg atgcggtatt
     6301 ttctccttac gcatctgtgc ggtatttcac accgcatacg cggatctgcg cagcaccatg
     6361 gcctgaaata acctctgaaa gaggaacttg gttaggtacc ttctgaggcg gaaagaacca
     6421 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     6481 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     6541 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     6601 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     6661 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     6721 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagcttaatt aactgttgac
     6781 aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc
     6841 caggaggcag atcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     6901 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     6961 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     7021 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     7081 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     7141 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     7201 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     7261 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     7321 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     7381 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     7441 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     7501 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     7561 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     7621 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     7681 gcgacgccca acctgccatc acgatggccg caataaaata tctttatttt cattacatct
     7741 gtgtgttggt tttttgtgtg aatcgatagc gataaggatc ctctttgcgc ttgcgttttc
     7801 ccttgtccag atagcccagt agctgacatt catccggggt cagcaccgtt tctgcggact
     7861 ggctttctac ccggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     7921 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     7981 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     8041 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     8101 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     8161 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     8221 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     8281 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     8341 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     8401 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     8461 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     8521 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     8581 atttcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     8641 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttata
     8701 gtccggaaat acaggaacgc acgctggatg gcccttcgct gggatggtga aaccatgaaa
     8761 aatggcagct tcagtggatt aagtgggggt aatgtggcct gtaccctctg gttgcatagg
     8821 tattcatacg gttaaaattt atcaggcgcg attgcggcag tttttcgggt ggtttgttgc
     8881 catttttacc tgtctgctgc cgtgatcgcg ctgaacgcgt tttagcggtg cgtacaatta
     8941 agggattatg gtaaatccac ttactgtctg ccctcgtagc catcgagata aaccgcagta
     9001 ctccggccac gatgcgtccg gcgtagagga tcgagatct
//
