LOCUS       sequence-476796-        7477 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express NSD3-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 7477)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7477)
  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            "Unpublished"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, NSD3-NanoLuc
FEATURES             Location/Qualifiers
     source          1..7477
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        253..271
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     253..270
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             2266..2778
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     polyA_signal    2955..3036
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2992..3011)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3053..4231
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3284..4222
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     4179..4199
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             4609..5403
                     /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(4663..4682)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5273..5292
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5427..5475
                     /note="synthetic polyadenylation signal"
     rep_origin      5601..6189
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6090..6109
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6361..7020)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     6935..6954
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(7052..7099)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     primer_bind     complement(7132..7152)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        join(7307..7477,1..133)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt
       61 cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt
      121 aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta
      181 ctattaataa ctagtggcgc gccagggagg cctatataag cagagctggt ttagtgaacc
      241 gtcagatcgc ccatttaggt gacactatag aatacaagct acttgttctt tttgcagcta
      301 gcgatcgcca tggatttctc tttctctttc atgcaaggga tcatgggaaa cacaattcag
      361 caaccacctc aactcattga ctccgccaac atccgtcagg aggatgcctt tgataacaac
      421 agtgacattg ctgaagatgg tggccagaca ccatatgaag ctactttgca gcaaggcttt
      481 cagtacccag ctacaacaga agatcttcct ccactcacaa atgggtatcc atcatcaatc
      541 agtgtgtatg aaactcaaac caaataccag tcatataatc agtatcctaa tgggtcagcc
      601 aatggctttg gtgcagttag aaactttagc cccactgact attatcattc agaaattcca
      661 aacacaagac cacatgaaat tctggaaaaa ccttcccctc cacagccacc acctcctcct
      721 tcggtaccac aaactgtgat tccaaagaag actggctcac ctgaaattaa actaaaaata
      781 accaaaacta tccagaatgg cagggaattg tttgagtctt ccctttgtgg agacctttta
      841 aatgaagtac aggcaagtga gcacacgaaa tcaaagcatg aaagcagaaa agaaaagagg
      901 aaaaaaagca acaagcatga ctcatcaaga tctgaagagc gcaagtcaca caaaatcccc
      961 aaattagaac cagaggaaca aaatagacca aatgagaggg ttgacactgt atcagaaaaa
     1021 ccaagggaag aaccagtact aaaagaggaa gccccagttc agccaatact atcttctgtt
     1081 ccaacaacgg aagtgtccac tggtgttaag tttcaggttg gcgatcttgt gtggtccaag
     1141 gtgggaacct atccttggtg gccttgtatg gtttcaagtg atccccagct tgaggttcat
     1201 actaaaatta acacaagagg tgcccgagaa tatcatgtcc agttttttag caaccagcca
     1261 gagagggcgt gggttcatga aaaacgggta cgagagtata aaggtcataa acagtatgaa
     1321 gaattactgg ctgaggcaac caaacaagcc agcaatcact ctgagaaaca aaagattcgg
     1381 aaaccccgac ctcagagaga acgtgctcag tgggatattg gcattgccca tgcagagaaa
     1441 gcattgaaaa tgactcgaga agaaagaata gaacagtata cttttattta cattgataaa
     1501 cagcctgaag aggctttatc ccaagcaaaa aagagtgttg cctccaaaac cgaagttaaa
     1561 aaaacccgac gaccaagatc tgtgctgaat actcagccag aacagaccaa tgcaggggag
     1621 gtggcctcct cactctcaag tactgaaatt cggagacata gccagaggcg gcacacaagt
     1681 gcggaagagg aagagccacc gcctgttaaa atagcctgga aaactgcggc agcaaggaaa
     1741 tccttaccag cttccattac gatgcacaaa gggagcctgg atttgcagaa gtgtaacatg
     1801 tctccagttg tgaaaattga acaagtgttt gctcttcaga atgctacagg ggatgggaaa
     1861 tttatcgatc aatttgttta ttcaacaaag ggaattggta acaaaacaga aataagtgtc
     1921 agggggcaag acaggcttat aatttctaca ccaaaccaga gaaatgaaaa gccaacgcag
     1981 agtgtatcat ctcctgaagc aacatctggt tctacaggct cagtagaaaa gaagcaacag
     2041 agaagatcaa ttagaactcg ttctgaatca gagaaatcca ctgaggttgt gccaaagaag
     2101 aagatcaaaa aggagcaggt tgaaacagtt cctcaggcta cagtgaagac tggattacag
     2161 aaagggtcgg cggaccgggg agtgcagggc tctgtcagat tcagtgacag ctccgtctcc
     2221 gcagcgattg aggaaactgt ggacgtttct ctcggctcga gcggcgtctt cacactcgaa
     2281 gatttcgttg gggactggcg acagacagcc ggctacaacc tggaccaagt ccttgaacag
     2341 ggaggtgtgt ccagtttgtt tcagaatctc ggggtgtccg taactccgat ccaaaggatt
     2401 gtcctgagcg gtgaaaatgg gctgaagatc gacatccatg tcatcatccc gtatgaaggt
     2461 ctgagcggcg accaaatggg ccagatcgaa aaaattttta aggtggtgta ccctgtggat
     2521 gatcatcact ttaaggtgat cctgcactat ggcacactgg taatcgacgg ggttacgccg
     2581 aacatgatcg actatttcgg acggccgtat gaaggcatcg ccgtgttcga cggcaaaaag
     2641 atcactgtaa cagggaccct gtggaacggc aacaaaatta tcgacgagcg cctgatcaac
     2701 cccgacggct ccctgctgtt ccgagtaacc atcaacggag tgaccggctg gcggctgtgc
     2761 gaacgcattc tggcgtaagg ccgcgactct agaaacacct aggctctcgt acgaagtcag
     2821 ctgataacgt cgacgtattg cggccgcgga tcataatcag ccataccaca tttgtagagg
     2881 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     2941 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     3001 tcacaaattt cacaaataaa gcattttttt cactgccgac gtcacgcgtc ttggctccgg
     3061 tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt
     3121 cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt
     3181 gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc
     3241 cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg
     3301 ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa ttacttccac
     3361 ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt gggagagttc
     3421 gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct ggcctgggcg
     3481 ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa
     3541 gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct ggcaagatag
     3601 tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg ccgcgggcgg
     3661 cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc
     3721 accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc ctggcctcgc
     3781 gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac cagttgcgtg
     3841 agcggaaaga tggccgcttc ccggccctgc tgcaggcagc tcaaaatgga ggacgcggcg
     3901 ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc cgtcctcagc
     3961 cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg attagttctc
     4021 gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga tggagtttcc
     4081 ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt aattctcctt
     4141 ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga cagtggttca
     4201 aagttttttt cttccatttc aggtgtcgtg aaagccgaat tctgcagata tccatcacac
     4261 tgtcgagcat gcatctagtc cagtgtggtg gaattcggct tcgtggggac gccgtacagg
     4321 gacgtgcacc tctcccgctg caccgcctcc agcgtcgccg ccggctcgaa ggacggggcc
     4381 gggatgacga tgcaggcggc gtgggaggtg gcgcccaagt tgcccatgac catgccgaag
     4441 cagtggtaga agggcaccgg cagacacacc cggtcctgct ccgtgtagcc gaccgtgcgg
     4501 cccacccagt agccgttgtt gaggatgttg tggtgggaga gcgtggcgcc cttggggaag
     4561 ccggtgtgcc ggaggtgtac tggatgttga ccgggaatcg atgccaccat gattgaacaa
     4621 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4681 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4741 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     4801 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     4861 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     4921 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4981 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     5041 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     5101 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     5161 gtcgtgaccc acggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     5221 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     5281 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     5341 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     5401 tgaactctag ccatcacgat ggccgcaata aaatatcttt attttcatta catctgtgtg
     5461 ttggtttttt gtgtgacgat tccggactat aaaaggatct aggtgaagat cctttttgat
     5521 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5581 gaaaagatca aaggatcttc ttgaaatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5641 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5701 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     5761 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5821 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5881 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5941 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6001 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6061 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6121 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6181 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6241 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     6301 gagtgagctg ataccggttg ggcgtcgctt ggtcggtcat ttcgaacccc agagtcccgc
     6361 ttacgccccg ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac
     6421 atggaagcca tcacagacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc
     6481 gccttgcgta taatatttgc ccatagtgaa aacgggggcg aagaagttgt ccatattggc
     6541 cacgtttaaa tcaaaactgg tgaaactcac ccagggattg gctgagacga aaaacatatt
     6601 ctcaataaac cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga
     6661 atatatgtgt agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt
     6721 ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc
     6781 accgtctttc attgccatac ggaattccgg atgagcattc atcaggcggg caagaatgtg
     6841 aataaaggcc ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat
     6901 atccagctga acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg
     6961 ttctttacga tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat
     7021 gatctgcctc ctgggtttag ttcctcacct tgtcgtatta tactatgccg atatactatg
     7081 ccgatgatta attgtcaaca gtattagaat tcgatttgac ctcccaccgt acacgcctac
     7141 cgcccatttg cgtcaacggg gcggggttat tacgacattt tggaaagtcc cgttgatttt
     7201 ggtgccaaaa caaactccca ttgacgtcaa tggggtggag acttggaaat ccccgtgagt
     7261 caaaccgcta tccacgccca ttggtgtact gccaaaaccg catcaccatg gtaatagcga
     7321 tgactaatac gtagatgtac tgccaagtag gaaagtcccg taaggtcatg tactgggcat
     7381 aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcggactt ggcatatgat
     7441 acacttgatg tactgccaag tgggcagttt accgtaa
//
