LOCUS       sequence-474974-        5135 bp    DNA     circular SYN 06-JUN-2025
DEFINITION  Express NanoLuc(R)-BRD9(BD) 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 5135)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 5135)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 6, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: NanoLuc-BRD9(BD) Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5135
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(902..920)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    957..1078
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(968..987)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1022..1041
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1323..1680
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1323..1514
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1346..1366)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1531..1666
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        1695..1742
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1774..2568
                     /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(1828..1847)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2438..2457
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2632..2680
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(2919..3507)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2999..3018)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    3623..3906
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     enhancer        4097..4476
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4477..4680
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4631..4651
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          4816..4948
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     4931..4950
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     4992..5011
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4992..5010
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(5024..5135,1..401)
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
ORIGIN
        1 tgtccgtaac tccgatccaa aggattgtcc tgagcggtga aaatgggctg aagatcgaca
       61 tccatgtcat catcccgtat gaaggtctga gcggcgacca aatgggccag atcgaaaaaa
      121 tttttaaggt ggtgtaccct gtggatgatc atcactttaa ggtgatcctg cactatggca
      181 cactggtaat cgacggggtt acgccgaaca tgatcgacta tttcggacgg ccgtatgaag
      241 gcatcgccgt gttcgacggc aaaaagatca ctgtaacagg gaccctgtgg aacggcaaca
      301 aaattatcga cgagcgcctg atcaaccccg acggctccct gctgttccga gtaaccatca
      361 acggagtgac cggctggcgg ctgtgcgaac gcattctggc gggctcgagc ggcgcgatcg
      421 ccatgccgcc cccagatcgg ccagtccgag cgtgccggac acagccagcc gaaaatgaga
      481 gcacacctat tcagcaactc ctggaacact tcctccgcca gcttcagaga aaagatcccc
      541 atggattttt tgcttttcct gtcacggatg caattgctcc tggatattca atgataataa
      601 aacatcccat ggattttggc accatgaaag acaaaattgt agctaatgaa tacaagtcag
      661 ttacggaatt taaggcagat ttcaagctga tgtgtgataa tgcaatgaca tacaataggc
      721 cagataccgt gtactacaag ttggcgaaga agatccttca cgcaggcttt aagatgatga
      781 gcaaacaggt ttaaacgaat tcgggctcgg tacccgggga tcctctagag tcgacctgca
      841 ggcatgcaag ctgatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
      901 accgctgagc aataactagc ataacccctt ggggcggccg cttcgagcag acatgataag
      961 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     1021 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     1081 caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggttttttt
     1141 aagcaagtaa aacctctaca aatgtggtaa aatcgaattt taacaaaata ttaacgctta
     1201 caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     1261 cggatctgcg cagcaccatg gcctgaaata acctctgaaa gaggaacttg gttaggtacc
     1321 ttctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     1381 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     1441 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     1501 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     1561 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     1621 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     1681 aagcttaatt aactgttgac aattaatcat cggcatagta tatcggcata gtataatacg
     1741 acaaggtgag gaactaaacc caggaggcag atcatgattg aacaagatgg attgcacgca
     1801 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     1861 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     1921 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     1981 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     2041 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     2101 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     2161 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     2221 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     2281 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     2341 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     2401 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     2461 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     2521 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     2581 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgatggccg caataaaata
     2641 tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagc gataaggatc
     2701 ctctttgcgc ttgcgttttc ccttgtccag atagcccagt agctgacatt catccggggt
     2761 cagcaccgtt tctgcggact ggctttctac ccggtatcag ctcactcaaa ggcggtaata
     2821 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     2881 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     2941 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3001 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3061 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3121 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3181 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3241 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3301 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     3361 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3421 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3481 attacgcgca gaaaaaaagg atttcaagaa gatcctttga tcttttctac ggggtctgac
     3541 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3601 ttcacctaga tccttttata gtccggaaat acaggaacgc acgctggatg gcccttcgct
     3661 gggatggtga aaccatgaaa aatggcagct tcagtggatt aagtgggggt aatgtggcct
     3721 gtaccctctg gttgcatagg tattcatacg gttaaaattt atcaggcgcg attgcggcag
     3781 tttttcgggt ggtttgttgc catttttacc tgtctgctgc cgtgatcgcg ctgaacgcgt
     3841 tttagcggtg cgtacaatta agggattatg gtaaatccac ttactgtctg ccctcgtagc
     3901 catcgagata aaccgcagta ctccggccac gatgcgtccg gcgtagagga tcgagatctt
     3961 caatattggc cattagccat attattcatt ggttatatag cataaatcaa tattggctat
     4021 tggccattgc atacgttgta tctatatcat aatatgtaca tttatattgg ctcatgtcca
     4081 atatgaccgc catgttggca ttgattattg actagttatt aatagtaatc aattacgggg
     4141 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     4201 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     4261 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     4321 cacttggcag tacatcaagt gtatcatatg ccaagtccgc cccctattga cgtcaatgac
     4381 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tacgggactt tcctacttgg
     4441 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacacc
     4501 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     4561 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taataacccc
     4621 gccccgttga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
     4681 ggtttagtga accgtcagat cactagaagc tttattgcgg tagtttatca cagttaaatt
     4741 gctaacgcag tcagtgcttc tgacacaaca gtctcgaact taagctgcag aagttggtcg
     4801 tgaggcactg ggcaggtaag tatcaaggtt acaagacagg tttaaggaga ccaatagaaa
     4861 ctgggcttgt cgagacagag aagactcttg cgtttctgat aggcacctat tggtcttact
     4921 gacatccact ttgcctttct ctccacaggt gtccactccc agttcaatta cagctcttaa
     4981 ggctagagta ttaatacgac tcactatagg gctagcgctc accatggtct tcacactcga
     5041 agatttcgtt ggggactggc gacagacagc cggctacaac ctggaccaag tccttgaaca
     5101 gggaggtgtg tccagtttgt ttcagaatct cgggg
//
