LOCUS       sequence-481504-        8385 bp    DNA     circular SYN 30-JUL-2025
DEFINITION  Express BRPF1-NanoLuc 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 8385)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8385)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 30, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: BRPF1-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..8385
                     /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             4728..5240
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(5328..5346)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    5383..5504
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5394..5413)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5448..5467
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5749..6106
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5749..5940
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5772..5792)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5957..6092
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        6121..6168
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6200..6994
                     /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(6254..6273)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6864..6883
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7058..7106
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(7345..7933)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7425..7444)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    8049..8332
                     /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 ggctagcgat cgccatgggg gtggactttg
     1081 atgtgaagac tttctgccac aacttgcggg cgactaagcc accatacgag tgcccggtgg
     1141 agacctgccg aaaggtctac aagagttaca gtggtattga gtaccacctg taccactatg
     1201 accacgacaa cccaccaccc ccacaacaaa ctccactccg caagcacaag aaaaaggggc
     1261 gccagtcacg cccagccaac aagcagtcac ccagcccctc agaggtctca cagtcaccag
     1321 gccgtgaggt gatgagctat gcacaggccc agcgtatggt ggaggtggac ttgcatggcc
     1381 gcgtccaccg catcagcatc tttgacaacc tggatgtggt gtcagaggat gaggaagccc
     1441 ccgaggaggc ccctgagaat ggcagcaaca aggagaacac tgagacacca gctgctactc
     1501 ccaagtcagg caaacataag aacaaggaga agcgcaagga ctccaaccat caccaccacc
     1561 acaatgtttc tgcgagcacc actcccaagc tgccagaggt ggtctatcgg gagctggaac
     1621 aggacacccc tgatgcccca ccccggccaa cttcctatta ccggtacatc gagaagtctg
     1681 cagaggagct ggacgaggaa gtagagtatg acatggacga ggaggactac atctggctgg
     1741 atatcatgaa tgagcgtcgg aagacagagg gtgtaagtcc catcccgcag gagatctttg
     1801 agtacctaat ggaccgactg gagaaggagt cgtactttga gagtcataat aaaggcgacc
     1861 ctaatgcgct agtggacgag gatgctgttt gctgtatctg caatgatggt gagtgccaga
     1921 acagcaatgt catcctcttc tgtgacatgt gcaacctggc cgtgcaccag gagtgctacg
     1981 gtgtccccta tatccctgag ggccagtggc tgtgccgccg ttgcctgcag tcaccctctc
     2041 gtgctgtgga ttgtgccctg tgccccaaca agggcggtgc cttcaagcag acagatgacg
     2101 ggcgctgggc ccatgtggtg tgtgccttgt ggatccctga ggtctgcttc gccaacacgg
     2161 tcttcctaga gcctattgac agcattgagc acatcccacc agctcgctgg aagctcacct
     2221 gctacatttg caaacaacgg ggctcagggg cctgcatcca gtgccacaag gccaactgtt
     2281 acacagcttt ccatgtgaca tgcgcccagc aggctggcct ttacatgaag atggagcctg
     2341 tgcgggagac aggcgccaac ggcacctctt tcagtgtccg caagacagcc tactgcgaca
     2401 tccacacgcc tccaggttca gcacgccgac tgcctgccct gtcccacagc gagggtgagg
     2461 aggatgaaga tgaggaggag gatgagggta agggctggag ctcagagaaa gtcaagaagg
     2521 ccaaggccaa gtcccggatc aaaatgaaga aggcacggaa gatcctggca gagaagcggg
     2581 cagcagcacc tgtggtgtca gtgccctgca tcccaccaca caggcttagt aaaatcacca
     2641 accgcctgac catccaaagg aagagccagt tcatgcagag gctgcacagc tactggacac
     2701 tgaagcggca gtcacggaat ggggtcccat tgctacgtcg cctgcagaca cacctgcaat
     2761 ctcagaggaa ctgtgaccaa gttgggagag attctgagga taagaactgg gcccttaaag
     2821 aacagctcaa gtcctggcag cggctccggc atgacttgga gcgagctcgg ctgctcgtgg
     2881 aattgatccg caagcgggaa aaactcaaaa gggagacgat caaggttcag cagattgcca
     2941 tggagatgca gctgactcct ttcctcatcc tccttcgcaa aaccttggag cagctccaag
     3001 agaaggacac aggcaacatc ttcagcgagc cggtccctct gtctgaggta cctgactacc
     3061 tagaccacat caaaaagccc atggactttt tcaccatgaa gcagaacttg gaggcttacc
     3121 gctacctgaa ttttgatgat tttgaggagg acttcaacct catcgtcagc aactgcctca
     3181 agtataacgc caaggacacc atcttctacc gggcagcagt gcggcttcgt gagcagggtg
     3241 gtgctgtgct ccgccaggcc cggcgccagg cagaaaaaat gggcattgac tttgagacgg
     3301 gcatgcatat cccccacagc ctggctggag atgaggccac acaccacact gaagatgcag
     3361 ccgaggaaga gcggctggtc ttgctggaga accagaagca cctgccagtg gaagaacagc
     3421 taaagctgct tctggagcgg ctggacgaag tgaatgccag caagcagagt gtgggccgct
     3481 cacggcgtgc aaagatgatc aagaaagaga tgacggcact gcggcggaag cttgcccatc
     3541 agcgagagac gggacgtgat ggccctgagc ggcatggccc ctcgagccgg ggtagtctga
     3601 caccccaccc ggcagcctgt gacaaggatg ggcagacaga tagtgcggca gaggagagca
     3661 gcagccagga gacaagcaaa ggcctgggtc ccaacatgtc ctcaaccccc gcacatgagg
     3721 tgggcaggag aacctcagtt ctgttctcca aaaagaaccc gaagacagct ggaccgccca
     3781 agaggccggg ccggcccccc aaaaaccggg agagccagat gacccccagc cacggaggca
     3841 gtcctgtggg gcccccccag ctccccatca tgagttccct gcgtcagcgc aagcggggta
     3901 ggagcccccg gcccagttcg agctcagaca gcgacagtga taagtccaca gaagaccccc
     3961 caatggactt accagccaat ggcttcagcg gtggaaacca accagtgaag aagagtttct
     4021 tggtataccg taatgactgc agccttcccc ggagcagctc agactctgag tccagcagca
     4081 gtagcagtag cagcgctgct tcagaccgga ccagcacaac gccctcaaaa caaggccggg
     4141 gcaaaccctc cttctctcgg ggcactttcc cagaggacag cagtgaggat acctcaggca
     4201 ctgagaatga ggcctactcc gtgggcactg gccgcggcgt gggccacagc atggtaagga
     4261 agagtctggg ccggggagct ggctggctgt cagaggatga ggactccccg ctggatgctc
     4321 tggacctcgt gtgggccaaa tgccgaggct atccatcata cccagctctg atcattgatc
     4381 caaagatgcc ccgagaaggt atgttccacc atggggttcc catccctgtg cccccactgg
     4441 aggtgctgaa acttggggag cagatgaccc aggaagcccg agagcatctc tacctcgtcc
     4501 tcttctttga caacaaacga acctggcagt ggctgcccag gaccaagctg gttcctctgg
     4561 gtgtgaacca ggacctagac aaggagaaga tgctggaggg ccgcaagtcc aacatccgca
     4621 agtcagtaca gatcgcctac cacagggctc tgcagcaccg cagcaaggtg caaggcgagc
     4681 agagcagtga gaccagcgat agtgatgttt ctctcggctc gagcggcgtc ttcacactcg
     4741 aagatttcgt tggggactgg cgacagacag ccggctacaa cctggaccaa gtccttgaac
     4801 agggaggtgt gtccagtttg tttcagaatc tcggggtgtc cgtaactccg atccaaagga
     4861 ttgtcctgag cggtgaaaat gggctgaaga tcgacatcca tgtcatcatc ccgtatgaag
     4921 gtctgagcgg cgaccaaatg ggccagatcg aaaaaatttt taaggtggtg taccctgtgg
     4981 atgatcatca ctttaaggtg atcctgcact atggcacact ggtaatcgac ggggttacgc
     5041 cgaacatgat cgactatttc ggacggccgt atgaaggcat cgccgtgttc gacggcaaaa
     5101 agatcactgt aacagggacc ctgtggaacg gcaacaaaat tatcgacgag cgcctgatca
     5161 accccgacgg ctccctgctg ttccgagtaa ccatcaacgg agtgaccggc tggcggctgt
     5221 gcgaacgcat tctggcgtaa ggccgcgact ctagagtcga cctgcaggca tgcaagctga
     5281 tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata
     5341 actagcataa ccccttgggg cggccgcttc gagcagacat gataagatac attgatgagt
     5401 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     5461 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     5521 ttcattttat gtttcaggtt cagggggaga tgtgggaggt ttttttaagc aagtaaaacc
     5581 tctacaaatg tggtaaaatc gaattttaac aaaatattaa cgcttacaat ttcctgatgc
     5641 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgcgga tctgcgcagc
     5701 accatggcct gaaataacct ctgaaagagg aacttggtta ggtaccttct gaggcggaaa
     5761 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     5821 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     5881 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     5941 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     6001 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     6061 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttaattaact
     6121 gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac
     6181 taaacccagg aggcagatca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     6241 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     6301 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     6361 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     6421 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     6481 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     6541 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     6601 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     6661 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     6721 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     6781 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     6841 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     6901 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     6961 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     7021 gaccaagcga cgcccaacct gccatcacga tggccgcaat aaaatatctt tattttcatt
     7081 acatctgtgt gttggttttt tgtgtgaatc gatagcgata aggatcctct ttgcgcttgc
     7141 gttttccctt gtccagatag cccagtagct gacattcatc cggggtcagc accgtttctg
     7201 cggactggct ttctacccgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     7261 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     7321 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     7381 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     7441 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     7501 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     7561 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     7621 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     7681 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     7741 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     7801 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     7861 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     7921 aaaaggattt caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     7981 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     8041 tttatagtcc ggaaatacag gaacgcacgc tggatggccc ttcgctggga tggtgaaacc
     8101 atgaaaaatg gcagcttcag tggattaagt gggggtaatg tggcctgtac cctctggttg
     8161 cataggtatt catacggtta aaatttatca ggcgcgattg cggcagtttt tcgggtggtt
     8221 tgttgccatt tttacctgtc tgctgccgtg atcgcgctga acgcgtttta gcggtgcgta
     8281 caattaaggg attatggtaa atccacttac tgtctgccct cgtagccatc gagataaacc
     8341 gcagtactcc ggccacgatg cgtccggcgt agaggatcga gatct
//
