LOCUS       sequence-479789-        6254 bp    DNA     circular SYN 04-AUG-2025
DEFINITION  Express NanoLuc-DCAF4 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 6254)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6254)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 4, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: NanoLuc-DCAF4 Vector
FEATURES             Location/Qualifiers
     source          1..6254
                     /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"
     primer_bind     complement(3197..3215)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3252..3373
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3263..3282)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3317..3336
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3618..3975
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3618..3809
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3641..3661)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3826..3961
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3990..4037
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4069..4863
                     /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(4123..4142)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4733..4752
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4927..4975
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5214..5802)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5294..5313)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5918..6201
                     /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 gaataaaagt cgctggcaga
     1621 gtagaagacg acatgggaga agaagccacc agcagaaccc ttggttcaga ctccgtgatt
     1681 ctgaagacag gtctgactcc cgggcagcac agcccgctca cgattccggc cacggtgatg
     1741 acgagtctcc gtcaacctcg tctggcacag ctgggacctc ctctgtgcca gagctacctg
     1801 ggttttactt tgaccctgaa aagaaacgct acttccgctt gctccctgga cataacaact
     1861 gcaaccccct gacgaaagag agcatccggc agaaggagat ggagagcaag agactgcggc
     1921 tgctccagga agaagacaga cggaaaaaga ttgccaggat gggatttaat gcatcttcca
     1981 tgctacgaaa aagccagctg ggttttctca acgtcaccaa ttactgccat ttagcccacg
     2041 agctgcgtct cagctgcatg gagaggaaaa aggtccagat tcgaagcatg gatccctccg
     2101 ccttggcaag cgaccgattt aacctcatac tggcagatac caacagtgac cggctcttca
     2161 cagtgaacga tgttaaagtt ggaggctcca agtatggtat catcaacctg caaagtctga
     2221 agacccctac gctcaaggtg ttcatgcacg aaaacctcta cttcaccaac cggaaggtga
     2281 attcggtgtg ctgggcctcg ctgaatcact tggattccca cattctgcta tgcctcatgg
     2341 gactcgcaga gactccaggc tgtgccaccc tgctcccagc atcactgttc gtcaatagtc
     2401 acccaggaat agaccggcct ggcatgctct gcagtttccg gatccctggt gcctggtcct
     2461 gtgcctggtc cctgaatatc caagcaaata actgcttcag tacaggcttg tctcggcggg
     2521 tcctgttgac caacgtggtg acgggacacc ggcagtcctt tgggaccaac agtgatgtct
     2581 tggcccagca gtttgctctc atggctcctc tgctgtttaa tggctgccgc tctggggaaa
     2641 tctttgccat tgatctgcgt tgtggaaatc aaggcaaggg atggaaggcc acccgcctgt
     2701 ttcatgattc agcagtgacc tctgtgcgga tcctccaaga tgagcaatac ctgatggctt
     2761 cagacatggc tggaaagatc aagctgtggg acctgaggac cacgaagtgc gtaaggcagt
     2821 acgaaggcca cgtgaatgag tacgcctacc tgcccctgca tgtgcacgag gaagaaggaa
     2881 tcctggtggc agtgggccag gactgctaca cgagaatctg gagcctccac gatgcccgcc
     2941 tactgagaac cataccctcc ccgtaccctg cctccaaggc cgacattccc agtgtggcct
     3001 tctcgtcgcg gctggggggc tcccggggcg cgccggggct gctcatggct gtcgggcagg
     3061 acctttactg ttactcctac agcgtttaaa cgaattcggg ctcggtaccc ggggatcctc
     3121 tagagtcgac ctgcaggcat gcaagctgat ccggctgcta acaaagcccg aaaggaagct
     3181 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ggccgcttcg
     3241 agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     3301 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     3361 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat
     3421 gtgggaggtt tttttaagca agtaaaacct ctacaaatgt ggtaaaatcg aattttaaca
     3481 aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     3541 ttcacaccgc atacgcggat ctgcgcagca ccatggcctg aaataacctc tgaaagagga
     3601 acttggttag gtaccttctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     3661 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3721 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3781 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3841 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3901 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3961 ctaggctttt gcaaaaagct taattaactg ttgacaatta atcatcggca tagtatatcg
     4021 gcatagtata atacgacaag gtgaggaact aaacccagga ggcagatcat gattgaacaa
     4081 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4141 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4201 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     4261 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     4321 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     4381 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4441 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4501 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4561 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     4621 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4681 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     4741 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     4801 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     4861 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgat
     4921 ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg
     4981 atagcgataa ggatcctctt tgcgcttgcg ttttcccttg tccagatagc ccagtagctg
     5041 acattcatcc ggggtcagca ccgtttctgc ggactggctt tctacccggt atcagctcac
     5101 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5161 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5221 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5281 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5341 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5401 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5461 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5521 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5581 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5641 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5701 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5761 gtttgcaagc agcagattac gcgcagaaaa aaaggatttc aagaagatcc tttgatcttt
     5821 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5881 ttatcaaaaa ggatcttcac ctagatcctt ttatagtccg gaaatacagg aacgcacgct
     5941 ggatggccct tcgctgggat ggtgaaacca tgaaaaatgg cagcttcagt ggattaagtg
     6001 ggggtaatgt ggcctgtacc ctctggttgc ataggtattc atacggttaa aatttatcag
     6061 gcgcgattgc ggcagttttt cgggtggttt gttgccattt ttacctgtct gctgccgtga
     6121 tcgcgctgaa cgcgttttag cggtgcgtac aattaaggga ttatggtaaa tccacttact
     6181 gtctgccctc gtagccatcg agataaaccg cagtactccg gccacgatgc gtccggcgta
     6241 gaggatcgag atct
//
