LOCUS       sequence-475206-        7246 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express NanoLuc-ENL 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 7246)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7246)
  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            "Unpublished"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-ENL Vector
FEATURES             Location/Qualifiers
     source          1..7246
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    17..98
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     42..61
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1999..2511)
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     promoter        complement(2550..2568)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2551..2568)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        2688..2991
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3146..3166
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        3199..3246
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3278..3937
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3344..3363)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      complement(4109..4697)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4189..4208)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4823..4871
                     /note="synthetic polyadenylation signal"
     CDS             complement(4895..5689)
                     /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(5006..5025)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5616..5635
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(6068..7246)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          6077..7015
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(6100..6120)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
ORIGIN
        1 aagacgcgtg acgtcggcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
       61 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
      121 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
      181 aatgtggtat ggctgattat gatccgcggc cgcaatacgt cgacgttatc agctgacttc
      241 gtacgagagc ctaggtgttt ctagaggatc cccgggtacc gagcccgaat tcgtttaaac
      301 tgtggccacg gcctccaggc agctctgcag tttgcgcacg gtggtctcgt ccagggagaa
      361 gaggtcgaag tcgaaggtgg tgttggtgac attgaagtgg ccagtctcct cgatcagatt
      421 cacaatctgc tgcagcacgt tgcgctcccg cagcgccatc agcctccggt gtagctccac
      481 cagctcatcc gtgtaggcct tgtcgtaggt gcccttcttg aggatcttct caggcttgct
      541 gcaggactcg gggctcctcc ggcctgacac cttgctgttg ggcgggggtg gcttctgggg
      601 gggtgggggc tcacggctgg gcagggagga gtcggcgctg ttgtcactct cgctgtcgct
      661 gaagctcaac ctggagtccc tccccgggtt ggtcttgccg gcagcctcct cgcctgacga
      721 agagtcgtcc tcgtcggact cctcggactg caggtcctcc accatggagc gcaggggtcc
      781 ttggctgtgg ttctgggatg gctcgaagtc tgagtctgag ctggagtctg agctggagct
      841 ggagttggac gggcttgact gggcagactc ggacttgaag gaggcctcgt cctctgagtt
      901 ggactcctcc acctccaggg cctttttggc ctcccggggc tcactctcgg ccttcacctt
      961 ctcccctctg gtgctgctct tgtccttggc cggcttcttg tccgagaagg aggaggagga
     1021 ggaggtgcgg ggcgaggtgc ctggagcact ccgggacccc ttcgagctgc tcttcttctg
     1081 cttcttggcg ctgggctttg gcgagtcggc ggtggccggc cgcttgctgg aagcccgggg
     1141 tgggggtggg ggtgggggtg ggggcccacc cttgggggac gtgctttcca gcttggtctc
     1201 tttcagggcc atcttgggtt ccttgaaggc agccttgggc ggtggcgcct tctcctcctt
     1261 gggcagccgg ccctcgccca gcttccgcga ggtgtccttg gagcttttgg cctgctcacg
     1321 ctccagctcc ttggaggagc tcttgctctc ggagtctttg cggggccgct cccggtgctc
     1381 cttggtcacc ttgtgtggct tggaggtctt gctgctctcc ttgttggcgt ccttggagcc
     1441 gtgggatggt ttggtcttct tggggtcaga gaaggcagag agtggaattg tgggtaacat
     1501 ggggtagtcg ggactgggcc tggacaccgt gtctgctcct tcgggcatta ccatcacccc
     1561 gccggcccgc aggagcttgt accggaactc cgtggtgggg ttgttgaagg tgagcttctc
     1621 gcagcgcagg tggttcacgg gcgggttgcc ttccaggttc aggaacaggt cgtaggtgaa
     1681 gcagaccttc ctcggctcct ccttgttttt gaagtgcacc tcgatgggca tgatgaagcc
     1741 agcgtacccc gactcctcta ctttgtaggg gggctccttg cacacgcgtc tgggcttggg
     1801 gaagctgtcg tgcagccaga agaccacctt ctccacgaag tgctggatgt cacattgctc
     1861 ggggccgcgg acaaacacca tccagtcgtg agtgaacccc tccgtggtgg gcttcttgcg
     1921 cagttgggcg cgatgcccca gctctaacct cacctggacg gtgcactgat tgtccatggc
     1981 gatcgcgccg ctcgagcccg ccagaatgcg ttcgcacagc cgccagccgg tcactccgtt
     2041 gatggttact cggaacagca gggagccgtc ggggttgatc aggcgctcgt cgataatttt
     2101 gttgccgttc cacagggtcc ctgttacagt gatctttttg ccgtcgaaca cggcgatgcc
     2161 ttcatacggc cgtccgaaat agtcgatcat gttcggcgta accccgtcga ttaccagtgt
     2221 gccatagtgc aggatcacct taaagtgatg atcatccaca gggtacacca ccttaaaaat
     2281 tttttcgatc tggcccattt ggtcgccgct cagaccttca tacgggatga tgacatggat
     2341 gtcgatcttc agcccatttt caccgctcag gacaatcctt tggatcggag ttacggacac
     2401 cccgagattc tgaaacaaac tggacacacc tccctgttca aggacttggt ccaggttgta
     2461 gccggctgtc tgtcgccagt ccccaacgaa atcttcgagt gtgaagacca tggtgagcgc
     2521 tagctgcaaa aagaacaagt agcttgtatt ctatagtgtc acctaaatgg gcgatctgac
     2581 ggttcactaa accagctctg cttatatagg cctccctggc gcgccactag ttattaatag
     2641 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     2701 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     2761 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     2821 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct
     2881 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg
     2941 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
     3001 ttttggcagt acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     3061 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
     3121 tgtcgtaata accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
     3181 aaatcgaatt ctaatactgt tgacaattaa tcatcggcat agtatatcgg catagtataa
     3241 tacgacaagg tgaggaacta aacccaggag gcagatcatg gagaaaaaaa tcactggata
     3301 taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt
     3361 tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt
     3421 aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa
     3481 tgctcatccg gaattccgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt
     3541 tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga
     3601 ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg
     3661 tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa
     3721 tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc
     3781 ccccgttttc actatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc
     3841 gattcaggtt catcatgccg tctgtgatgg cttccatgtc ggcagaatgc ttaatgaatt
     3901 acaacagtac tgcgatgagt ggcagggcgg ggcgtaagcg ggactctggg gttcgaaatg
     3961 accgaccaag cgacgcccaa ccggtatcag ctcactcaaa ggcggtaata cggttatcca
     4021 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4081 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4141 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4201 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4261 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4321 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4381 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4441 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4501 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     4561 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4621 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4681 gaaaaaaagg atttcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4741 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4801 tccttttata gtccggaatc gtcacacaaa aaaccaacac acagatgtaa tgaaaataaa
     4861 gatattttat tgcggccatc gtgatggcta gagttcagaa gaactcgtca agaaggcgat
     4921 agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag
     4981 cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc
     5041 ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca
     5101 tgatattcgg caagcaggca tcgccgtggg tcacgacgag atcctcgccg tcgggcatgc
     5161 gcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat
     5221 catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg
     5281 cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag
     5341 ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca
     5401 cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc
     5461 aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc agttcattca
     5521 gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga
     5581 acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct
     5641 ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg gtggcatcga
     5701 ttcccggtca acatccagta cacctccggc accaccggct tccccaaggg cgccacgctc
     5761 tcccaccaca acatcctcaa caacggctac tgggtgggcc gcacggtcgg ctacacggag
     5821 caggaccggg tgtgtctgcc ggtgcccttc taccactgct tcggcatggt catgggcaac
     5881 ttgggcgcca cctcccacgc cgcctgcatc gtcatcccgg ccccgtcctt cgagccggcg
     5941 gcgacgctgg aggcggtgca gcgggagagg tgcacgtccc tgtacggcgt ccccacgaag
     6001 ccgaattcca ccacactgga ctagatgcat gctcgacagt gtgatggata tctgcagaat
     6061 tcggctttca cgacacctga aatggaagaa aaaaactttg aaccactgtc tgaggcttga
     6121 gaatgaacca agatccaaac tcaaaaaggg caaattccaa ggagaattac atcaagtgcc
     6181 aagctggcct aacttcagtc tccacccact cagtgtgggg aaactccatc gcataaaacc
     6241 cctcccccca acctaaagac gacgtactcc aaaagctcga gaactaatcg aggtgcctgg
     6301 acggcgcccg gtactccgtg gagtcacatg aagcgacggc tgaggacgga aaggcccttt
     6361 tcctttgtgt gggtgactca cccgcccgct ctcccgagcg ccgcgtcctc cattttgagc
     6421 tgcctgcagc agggccggga agcggccatc tttccgctca cgcaactggt gccgaccggg
     6481 ccagccttgc cgcccagggc ggggcgatac acggcggcgc gaggccaggc accagagcag
     6541 gccggccagc ttgagactac ccccgtccga ttctcggtgg ccgcgctcgc aggccccgcc
     6601 tcgccgaaca tgtgcgctgg gacgcacggg ccccgtcgcc gcccgcggcc ccaaaaaccg
     6661 aaataccagt gtgcagatct tggcccgcat ttacaagact atcttgccag aaaaaaagcg
     6721 tcgcagcagg tcatcaaaaa ttttaaatgg ctagagactt atcgaaagca gcgagacagg
     6781 cgcgaaggtg ccaccagatt cgcacgcggc ggccccagcg cccaggccag gcctcaactc
     6841 aagcacgagg cgaaggggct ccttaagcgc aaggcctcga actctcccac ccacttccaa
     6901 cccgaagctc gggatcaaga atcacgtact gcagccaggt ggaagtaatt caaggcacgc
     6961 aagggccata acccgtaaag aggccaggcc cgcgggaacc acacacggca cttacctgtg
     7021 ttctggcggc aaacccgttg cgaaaaagaa cgttcacggc gactactgca cttatatacg
     7081 gttctccccc accctcggga aaaaggcgga gccagtacac gacatcactt tcccagttta
     7141 ccccgcgcca ccttctctag gcaccggttc aattgccgac ccctcccccc aacttctcgg
     7201 ggactgtggg cgatgtgcgc tctgcccact gacgggcacc ggagcc
//
