LOCUS       sequence-479807-        6560 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express NanoLuc-BRD9 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 6560)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6560)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 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            "Unpublished"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-BRD9
FEATURES             Location/Qualifiers
     source          1..6560
                     /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             1065..1577
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             2894..2905
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(3503..3521)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3558..3679
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3569..3588)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3623..3642
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3924..4281
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3924..4115
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3947..3967)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4132..4267
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4296..4343
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4375..5169
                     /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(4429..4448)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5039..5058
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5233..5281
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5520..6108)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5600..5619)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6224..6507
                     /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 ggctagcgct caccatggtc ttcacactcg
     1081 aagatttcgt tggggactgg cgacagacag ccggctacaa cctggaccaa gtccttgaac
     1141 agggaggtgt gtccagtttg tttcagaatc tcggggtgtc cgtaactccg atccaaagga
     1201 ttgtcctgag cggtgaaaat gggctgaaga tcgacatcca tgtcatcatc ccgtatgaag
     1261 gtctgagcgg cgaccaaatg ggccagatcg aaaaaatttt taaggtggtg taccctgtgg
     1321 atgatcatca ctttaaggtg atcctgcact atggcacact ggtaatcgac ggggttacgc
     1381 cgaacatgat cgactatttc ggacggccgt atgaaggcat cgccgtgttc gacggcaaaa
     1441 agatcactgt aacagggacc ctgtggaacg gcaacaaaat tatcgacgag cgcctgatca
     1501 accccgacgg ctccctgctg ttccgagtaa ccatcaacgg agtgaccggc tggcggctgt
     1561 gcgaacgcat tctggcgggc tcgagcggcg cgatcgccat gggcaagaag cacaagaagc
     1621 acaaggccga gtggcgctcg tcctacgagg attatgccga caagcccctg gagaagcctc
     1681 taaagctagt cctgaaggtc ggaggaagtg aagtgactga actctcagga tccggccacg
     1741 actccagtta ctatgatgac aggtcagacc atgagcgaga gaggcacaaa gaaaagaaaa
     1801 agaagaagaa gaagaagtcc gagaaggaga agcatctgga cgatgaggaa agaaggaagc
     1861 gaaaggaaga gaagaagcgg aagcgagaga gggagcactg tgacacggag ggagaggctg
     1921 acgactttga tcctgggaag aaggtggagg tggagccgcc cccagatcgg ccagtccgag
     1981 cgtgccggac acagccagcc gaaaatgaga gcacacctat tcagcaactc ctggaacact
     2041 tcctccgcca gcttcagaga aaagatcccc atggattttt tgcttttcct gtcacggatg
     2101 caattgctcc tggatattca atgataataa aacatcccat ggattttggc accatgaaag
     2161 acaaaattgt agctaatgaa tacaagtcag ttacggaatt taaggcagat ttcaagctga
     2221 tgtgtgataa tgcaatgaca tacaataggc cagataccgt gtactacaag ttggcgaaga
     2281 agatccttca cgcaggcttt aagatgatga gcaaacaggc agctcttttg ggcaatgaag
     2341 atacagctgt tgaggaacct gtccctgaag ttgtaccagt acaagtagaa actgccaaga
     2401 aatccaaaaa gccgagtaga gaagttatca gctgcatgtt tgagcctgaa gggaatgcct
     2461 gcagcttgac ggacagtacc gcagaggagc acgtgctggc gctggtggag cacgcagctg
     2521 acgaagctcg ggacaggatc aaccggttcc tcccaggcgg caagatgggc tatctgaaga
     2581 ggaacgggga cgggagcctg ctctacagcg tggtcaacac ggccgagccg gacgctgatg
     2641 aggaggagac ccacccggtg gacttgagct cgctctccag taagctactc ccaggcttca
     2701 ccacgctggg cttcaaagac gagagaagaa acaaagtcac ctttctctcc agtgccacta
     2761 ctgcgctttc gatgcagaat aattcagtat ttggcgactt gaagtcggac gagatggagc
     2821 tgctctactc agcctacgga gatgagacag gcgtgcagtg tgcgctgagc ctgcaggagt
     2881 ttgtgaagga tgctgggagc tacagcaaga aagtggtgga cgacctcctg gaccagatca
     2941 caggcggaga ccactctagg acgctcttcc agctgaagca gagaagaaat gttcccatga
     3001 agcctccaga tgaagccaag gttggggaca ccctaggaga cagcagcagc tctgttctgg
     3061 agttcatgtc gatgaagtcc tatcccgacg tttctgtgga tatctccatg ctcagctctc
     3121 tggggaaggt gaagaaggag ctggaccctg acgacagcca tttgaacttg gatgagacga
     3181 cgaagctcct gcaggacctg cacgaagcac aggcggagcg cggcggctct cggccgtcgt
     3241 ccaacctcag ctccctgtcc aacgcctccg agagggacca gcaccacctg ggaagccctt
     3301 ctcgcctgag tgtcggggag cagccagacg tcacccacga cccctatgag tttcttcagt
     3361 ctccagagcc tgcggcctct gccaagaccg tttaaacgaa ttcgggctcg gtacccgggg
     3421 atcctctaga gtcgacctgc aggcatgcaa gctgatccgg ctgctaacaa agcccgaaag
     3481 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcggcc
     3541 gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     3601 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     3661 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     3721 ggagatgtgg gaggtttttt taagcaagta aaacctctac aaatgtggta aaatcgaatt
     3781 ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg
     3841 cggtatttca caccgcatac gcggatctgc gcagcaccat ggcctgaaat aacctctgaa
     3901 agaggaactt ggttaggtac cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     3961 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4021 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4081 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     4141 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4201 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     4261 ggaggcctag gcttttgcaa aaagcttaat taactgttga caattaatca tcggcatagt
     4321 atatcggcat agtataatac gacaaggtga ggaactaaac ccaggaggca gatcatgatt
     4381 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4441 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4501 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     4561 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4621 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4681 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4741 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4801 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4861 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4921 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4981 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     5041 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     5101 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     5161 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     5221 cacgatggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt
     5281 gaatcgatag cgataaggat cctctttgcg cttgcgtttt cccttgtcca gatagcccag
     5341 tagctgacat tcatccgggg tcagcaccgt ttctgcggac tggctttcta cccggtatca
     5401 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5461 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5521 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5581 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5641 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5701 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5761 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5821 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5881 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5941 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     6001 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     6061 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatttcaaga agatcctttg
     6121 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     6181 atgagattat caaaaaggat cttcacctag atccttttat agtccggaaa tacaggaacg
     6241 cacgctggat ggcccttcgc tgggatggtg aaaccatgaa aaatggcagc ttcagtggat
     6301 taagtggggg taatgtggcc tgtaccctct ggttgcatag gtattcatac ggttaaaatt
     6361 tatcaggcgc gattgcggca gtttttcggg tggtttgttg ccatttttac ctgtctgctg
     6421 ccgtgatcgc gctgaacgcg ttttagcggt gcgtacaatt aagggattat ggtaaatcca
     6481 cttactgtct gccctcgtag ccatcgagat aaaccgcagt actccggcca cgatgcgtcc
     6541 ggcgtagagg atcgagatct
//
