LOCUS       sequence-470720-        7504 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express NanoLuc(R)-NSD3 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 7504)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7504)
  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            "Unpublished"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: NanoBRET Assay Vector, NanoLuc-NSD3
FEATURES             Location/Qualifiers
     source          1..7504
                     /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(2257..2769)
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     promoter        complement(2808..2826)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2809..2826)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        2946..3249
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3404..3424
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        3457..3504
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3536..4195
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3602..3621)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      complement(4367..4955)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4447..4466)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5081..5129
                     /note="synthetic polyadenylation signal"
     CDS             complement(5153..5947)
                     /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(5264..5283)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5874..5893
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(6326..7504)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          6335..7273
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(6358..6378)
                     /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 gtccacagtt tcctcaatcg ctgcggagac ggagctgtca ctgaatctga cagagccctg
      361 cactccccgg tccgccgacc ctttctgtaa tccagtcttc actgtagcct gaggaactgt
      421 ttcaacctgc tcctttttga tcttcttctt tggcacaacc tcagtggatt tctctgattc
      481 agaacgagtt ctaattgatc ttctctgttg cttcttttct actgagcctg tagaaccaga
      541 tgttgcttca ggagatgata cactctgcgt tggcttttca tttctctggt ttggtgtaga
      601 aattataagc ctgtcttgcc ccctgacact tatttctgtt ttgttaccaa ttccctttgt
      661 tgaataaaca aattgatcga taaatttccc atcccctgta gcattctgaa gagcaaacac
      721 ttgttcaatt ttcacaactg gagacatgtt acacttctgc aaatccaggc tccctttgtg
      781 catcgtaatg gaagctggta aggatttcct tgctgccgca gttttccagg ctattttaac
      841 aggcggtggc tcttcctctt ccgcacttgt gtgccgcctc tggctatgtc tccgaatttc
      901 agtacttgag agtgaggagg ccacctcccc tgcattggtc tgttctggct gagtattcag
      961 cacagatctt ggtcgtcggg tttttttaac ttcggttttg gaggcaacac tcttttttgc
     1021 ttgggataaa gcctcttcag gctgtttatc aatgtaaata aaagtatact gttctattct
     1081 ttcttctcga gtcattttca atgctttctc tgcatgggca atgccaatat cccactgagc
     1141 acgttctctc tgaggtcggg gtttccgaat cttttgtttc tcagagtgat tgctggcttg
     1201 tttggttgcc tcagccagta attcttcata ctgtttatga cctttatact ctcgtacccg
     1261 tttttcatga acccacgccc tctctggctg gttgctaaaa aactggacat gatattctcg
     1321 ggcacctctt gtgttaattt tagtatgaac ctcaagctgg ggatcacttg aaaccataca
     1381 aggccaccaa ggataggttc ccaccttgga ccacacaaga tcgccaacct gaaacttaac
     1441 accagtggac acttccgttg ttggaacaga agatagtatt ggctgaactg gggcttcctc
     1501 ttttagtact ggttcttccc ttggtttttc tgatacagtg tcaaccctct catttggtct
     1561 attttgttcc tctggttcta atttggggat tttgtgtgac ttgcgctctt cagatcttga
     1621 tgagtcatgc ttgttgcttt ttttcctctt ttcttttctg ctttcatgct ttgatttcgt
     1681 gtgctcactt gcctgtactt catttaaaag gtctccacaa agggaagact caaacaattc
     1741 cctgccattc tggatagttt tggttatttt tagtttaatt tcaggtgagc cagtcttctt
     1801 tggaatcaca gtttgtggta ccgaaggagg aggtggtggc tgtggagggg aaggtttttc
     1861 cagaatttca tgtggtcttg tgtttggaat ttctgaatga taatagtcag tggggctaaa
     1921 gtttctaact gcaccaaagc cattggctga cccattagga tactgattat atgactggta
     1981 tttggtttga gtttcataca cactgattga tgatggatac ccatttgtga gtggaggaag
     2041 atcttctgtt gtagctgggt actgaaagcc ttgctgcaaa gtagcttcat atggtgtctg
     2101 gccaccatct tcagcaatgt cactgttgtt atcaaaggca tcctcctgac ggatgttggc
     2161 ggagtcaatg agttgaggtg gttgctgaat tgtgtttccc atgatccctt gcatgaaaga
     2221 gaaagagaaa tccatggcga tcgcgccgct cgagcccgcc agaatgcgtt cgcacagccg
     2281 ccagccggtc actccgttga tggttactcg gaacagcagg gagccgtcgg ggttgatcag
     2341 gcgctcgtcg ataattttgt tgccgttcca cagggtccct gttacagtga tctttttgcc
     2401 gtcgaacacg gcgatgcctt catacggccg tccgaaatag tcgatcatgt tcggcgtaac
     2461 cccgtcgatt accagtgtgc catagtgcag gatcacctta aagtgatgat catccacagg
     2521 gtacaccacc ttaaaaattt tttcgatctg gcccatttgg tcgccgctca gaccttcata
     2581 cgggatgatg acatggatgt cgatcttcag cccattttca ccgctcagga caatcctttg
     2641 gatcggagtt acggacaccc cgagattctg aaacaaactg gacacacctc cctgttcaag
     2701 gacttggtcc aggttgtagc cggctgtctg tcgccagtcc ccaacgaaat cttcgagtgt
     2761 gaagaccatg gtgagcgcta gctgcaaaaa gaacaagtag cttgtattct atagtgtcac
     2821 ctaaatgggc gatctgacgg ttcactaaac cagctctgct tatataggcc tccctggcgc
     2881 gccactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     2941 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     3001 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     3061 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     3121 atgccaagtc cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     3181 cagtacatga ccttacggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     3241 attaccatgg tgatgcggtt ttggcagtac accaatgggc gtggatagcg gtttgactca
     3301 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
     3361 caacgggact ttccaaaatg tcgtaataac cccgccccgt tgacgcaaat gggcggtagg
     3421 cgtgtacggt gggaggtcaa atcgaattct aatactgttg acaattaatc atcggcatag
     3481 tatatcggca tagtataata cgacaaggtg aggaactaaa cccaggaggc agatcatgga
     3541 gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt
     3601 tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac
     3661 ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat
     3721 tcttgcccgc ctgatgaatg ctcatccgga attccgtatg gcaatgaaag acggtgagct
     3781 ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt
     3841 ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca
     3901 agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat
     3961 gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa
     4021 tatggacaac ttcttcgccc ccgttttcac tatgggcaaa tattatacgc aaggcgacaa
     4081 ggtgctgatg ccgctggcga ttcaggttca tcatgccgtc tgtgatggct tccatgtcgg
     4141 cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaagcggg
     4201 actctggggt tcgaaatgac cgaccaagcg acgcccaacc ggtatcagct cactcaaagg
     4261 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4321 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4381 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4441 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4501 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4561 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4621 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4681 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4741 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4801 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4861 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4921 agcagcagat tacgcgcaga aaaaaaggat ttcaagaaga tcctttgatc ttttctacgg
     4981 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5041 aaaggatctt cacctagatc cttttatagt ccggaatcgt cacacaaaaa accaacacac
     5101 agatgtaatg aaaataaaga tattttattg cggccatcgt gatggctaga gttcagaaga
     5161 actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa
     5221 gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca
     5281 acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa
     5341 agcggccatt ttccaccatg atattcggca agcaggcatc gccgtgggtc acgacgagat
     5401 cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag ttcggctggc gcgagcccct
     5461 gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc
     5521 gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca
     5581 gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca
     5641 ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa
     5701 cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct
     5761 cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc
     5821 cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt
     5881 catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt
     5941 caatcatggt ggcatcgatt cccggtcaac atccagtaca cctccggcac caccggcttc
     6001 cccaagggcg ccacgctctc ccaccacaac atcctcaaca acggctactg ggtgggccgc
     6061 acggtcggct acacggagca ggaccgggtg tgtctgccgg tgcccttcta ccactgcttc
     6121 ggcatggtca tgggcaactt gggcgccacc tcccacgccg cctgcatcgt catcccggcc
     6181 ccgtccttcg agccggcggc gacgctggag gcggtgcagc gggagaggtg cacgtccctg
     6241 tacggcgtcc ccacgaagcc gaattccacc acactggact agatgcatgc tcgacagtgt
     6301 gatggatatc tgcagaattc ggctttcacg acacctgaaa tggaagaaaa aaactttgaa
     6361 ccactgtctg aggcttgaga atgaaccaag atccaaactc aaaaagggca aattccaagg
     6421 agaattacat caagtgccaa gctggcctaa cttcagtctc cacccactca gtgtggggaa
     6481 actccatcgc ataaaacccc tccccccaac ctaaagacga cgtactccaa aagctcgaga
     6541 actaatcgag gtgcctggac ggcgcccggt actccgtgga gtcacatgaa gcgacggctg
     6601 aggacggaaa ggcccttttc ctttgtgtgg gtgactcacc cgcccgctct cccgagcgcc
     6661 gcgtcctcca ttttgagctg cctgcagcag ggccgggaag cggccatctt tccgctcacg
     6721 caactggtgc cgaccgggcc agccttgccg cccagggcgg ggcgatacac ggcggcgcga
     6781 ggccaggcac cagagcaggc cggccagctt gagactaccc ccgtccgatt ctcggtggcc
     6841 gcgctcgcag gccccgcctc gccgaacatg tgcgctggga cgcacgggcc ccgtcgccgc
     6901 ccgcggcccc aaaaaccgaa ataccagtgt gcagatcttg gcccgcattt acaagactat
     6961 cttgccagaa aaaaagcgtc gcagcaggtc atcaaaaatt ttaaatggct agagacttat
     7021 cgaaagcagc gagacaggcg cgaaggtgcc accagattcg cacgcggcgg ccccagcgcc
     7081 caggccaggc ctcaactcaa gcacgaggcg aaggggctcc ttaagcgcaa ggcctcgaac
     7141 tctcccaccc acttccaacc cgaagctcgg gatcaagaat cacgtactgc agccaggtgg
     7201 aagtaattca aggcacgcaa gggccataac ccgtaaagag gccaggcccg cgggaaccac
     7261 acacggcact tacctgtgtt ctggcggcaa acccgttgcg aaaaagaacg ttcacggcga
     7321 ctactgcact tatatacggt tctcccccac cctcgggaaa aaggcggagc cagtacacga
     7381 catcactttc ccagtttacc ccgcgccacc ttctctaggc accggttcaa ttgccgaccc
     7441 ctccccccaa cttctcgggg actgtgggcg atgtgcgctc tgcccactga cgggcaccgg
     7501 agcc
//
