LOCUS       sequence-472466-        5159 bp    DNA     circular SYN 13-MAY-2025
DEFINITION  Express NanoLuc(R)-BRPF1(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 5159)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 5159)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: NanoLuc-BRPF1(BD) Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5159
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        395..774
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        775..978
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     929..949
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          1114..1246
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1229..1248
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1290..1309
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1290..1308
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1322..1834
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     primer_bind     complement(2359..2377)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2414..2535
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2425..2444)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2479..2498
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2780..3137
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2780..2971
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2803..2823)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2988..3123
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3152..3199
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3231..4025
                     /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(3285..3304)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3895..3914
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4089..4137
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4376..4964)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4456..4475)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    join(5080..5159,1..204)
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 gtgggggtaa tgtggcctgt accctctggt tgcataggta ttcatacggt taaaatttat
       61 caggcgcgat tgcggcagtt tttcgggtgg tttgttgcca tttttacctg tctgctgccg
      121 tgatcgcgct gaacgcgttt tagcggtgcg tacaattaag ggattatggt aaatccactt
      181 actgtctgcc ctcgtagcca tcgagataaa ccgcagtact ccggccacga tgcgtccggc
      241 gtagaggatc gagatcttca atattggcca ttagccatat tattcattgg ttatatagca
      301 taaatcaata ttggctattg gccattgcat acgttgtatc tatatcataa tatgtacatt
      361 tatattggct catgtccaat atgaccgcca tgttggcatt gattattgac tagttattaa
      421 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
      481 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      541 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      601 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtccgccc
      661 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
      721 cgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
      781 cggttttggc agtacaccaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
      841 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
      901 aaatgtcgta ataaccccgc cccgttgacg caaatgggcg gtaggcgtgt acggtgggag
      961 gtctatataa gcagagctgg tttagtgaac cgtcagatca ctagaagctt tattgcggta
     1021 gtttatcaca gttaaattgc taacgcagtc agtgcttctg acacaacagt ctcgaactta
     1081 agctgcagaa gttggtcgtg aggcactggg caggtaagta tcaaggttac aagacaggtt
     1141 taaggagacc aatagaaact gggcttgtcg agacagagaa gactcttgcg tttctgatag
     1201 gcacctattg gtcttactga catccacttt gcctttctct ccacaggtgt ccactcccag
     1261 ttcaattaca gctcttaagg ctagagtatt aatacgactc actatagggc tagcgctcac
     1321 catggtcttc acactcgaag atttcgttgg ggactggcga cagacagccg gctacaacct
     1381 ggaccaagtc cttgaacagg gaggtgtgtc cagtttgttt cagaatctcg gggtgtccgt
     1441 aactccgatc caaaggattg tcctgagcgg tgaaaatggg ctgaagatcg acatccatgt
     1501 catcatcccg tatgaaggtc tgagcggcga ccaaatgggc cagatcgaaa aaatttttaa
     1561 ggtggtgtac cctgtggatg atcatcactt taaggtgatc ctgcactatg gcacactggt
     1621 aatcgacggg gttacgccga acatgatcga ctatttcgga cggccgtatg aaggcatcgc
     1681 cgtgttcgac ggcaaaaaga tcactgtaac agggaccctg tggaacggca acaaaattat
     1741 cgacgagcgc ctgatcaacc ccgacggctc cctgctgttc cgagtaacca tcaacggagt
     1801 gaccggctgg cggctgtgcg aacgcattct ggcgggctcg agcggcgcga tcgccatggt
     1861 tcagcagatt gccatggaga tgcagctgac tcctttcctc atcctccttc gcaaaacctt
     1921 ggagcagctc caagagaagg acacaggcaa catcttcagc gagccggtcc ctctgtctga
     1981 ggtacctgac tacctagacc acatcaaaaa gcccatggac tttttcacca tgaagcagaa
     2041 cttggaggct taccgctacc tgaattttga tgattttgag gaggacttca acctcatcgt
     2101 cagcaactgc ctcaagtata acgccaagga caccatcttc taccgggcag cagtgcggct
     2161 tcgtgagcag ggtggtgctg tgctccgcca ggcccggcgc caggcagaaa aaatgggcat
     2221 tgactttgag acgggcatgc atatcgttta aacgaattcg ggctcggtac ccggggatcc
     2281 tctagagtcg acctgcaggc atgcaagctg atccggctgc taacaaagcc cgaaaggaag
     2341 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcggccgctt
     2401 cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     2461 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     2521 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     2581 atgtgggagg tttttttaag caagtaaaac ctctacaaat gtggtaaaat cgaattttaa
     2641 caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt
     2701 atttcacacc gcatacgcgg atctgcgcag caccatggcc tgaaataacc tctgaaagag
     2761 gaacttggtt aggtaccttc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     2821 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     2881 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2941 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3001 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3061 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3121 gcctaggctt ttgcaaaaag cttaattaac tgttgacaat taatcatcgg catagtatat
     3181 cggcatagta taatacgaca aggtgaggaa ctaaacccag gaggcagatc atgattgaac
     3241 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3301 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3361 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     3421 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     3481 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     3541 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     3601 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     3661 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     3721 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     3781 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     3841 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     3901 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     3961 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4021 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4081 atggccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgaat
     4141 cgatagcgat aaggatcctc tttgcgcttg cgttttccct tgtccagata gcccagtagc
     4201 tgacattcat ccggggtcag caccgtttct gcggactggc tttctacccg gtatcagctc
     4261 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     4321 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     4381 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     4441 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     4501 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4561 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4621 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4681 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4741 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4801 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4861 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4921 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatt tcaagaagat cctttgatct
     4981 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     5041 gattatcaaa aaggatcttc acctagatcc ttttatagtc cggaaataca ggaacgcacg
     5101 ctggatggcc cttcgctggg atggtgaaac catgaaaaat ggcagcttca gtggattaa
//
