LOCUS       sequence-481955-        5881 bp    DNA     circular SYN 04-AUG-2025
DEFINITION  Express DAP12-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 5881)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5881)
  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            "Unpublished"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: NanoBRET Assay Vector, DAP12-NanoLuc
FEATURES             Location/Qualifiers
     source          1..5881
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    17..98
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     42..61
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(275..787)
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     promoter        complement(1186..1204)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1187..1204)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        1324..1627
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1782..1802
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        1835..1882
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1914..2573
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1980..1999)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      complement(2745..3333)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2825..2844)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3459..3507
                     /note="synthetic polyadenylation signal"
     CDS             complement(3531..4325)
                     /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(3642..3661)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4252..4271
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4703..5881)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          4712..5650
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(4735..4755)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
ORIGIN
        1 aagacgcgtg acgtcggcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
       61 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
      121 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
      181 aatgtggtat ggctgattat gatccgcggc cgcaatacgt cgacgttatc agctgacttc
      241 gtacgagagc ctaggtgttt ctagagtcgc ggccttacgc cagaatgcgt tcgcacagcc
      301 gccagccggt cactccgttg atggttactc ggaacagcag ggagccgtcg gggttgatca
      361 ggcgctcgtc gataattttg ttgccgttcc acagggtccc tgttacagtg atctttttgc
      421 cgtcgaacac ggcgatgcct tcatacggcc gtccgaaata gtcgatcatg ttcggcgtaa
      481 ccccgtcgat taccagtgtg ccatagtgca ggatcacctt aaagtgatga tcatccacag
      541 ggtacaccac cttaaaaatt ttttcgatct ggcccatttg gtcgccgctc agaccttcat
      601 acgggatgat gacatggatg tcgatcttca gcccattttc accgctcagg acaatccttt
      661 ggatcggagt tacggacacc ccgagattct gaaacaaact ggacacacct ccctgttcaa
      721 ggacttggtc caggttgtag ccggctgtct gtcgccagtc cccaacgaaa tcttcgagtg
      781 tgaagacgcc gctcgagccg agagaaactt tgtaatacgg cctctgtgtg ttgaggtcgc
      841 tgtagacatc cgacctctga ccctggagct cctgataagg cgactcggtc tcagtgatac
      901 gctgtttccg ggtcgctgcc tccgcagccc ctcgcccccg agggaccagc cggcccagga
      961 agtacacggc cagggcaatg agcactgtca gcaccaggtc tcccatcacg atccctgcca
     1021 gcacgcccgg gctcaccgta gagcaactgc aatcgctctg ggcctgggcc tggacaggac
     1081 ggagaccact tacagccagc aggagaggca ggagcaggag cctgctgcag ggttcaagtc
     1141 cccccatggc gatcgctagc tgcaaaaaga acaagtagct tgtattctat agtgtcacct
     1201 aaatgggcga tctgacggtt cactaaacca gctctgctta tataggcctc cctggcgcgc
     1261 cactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
     1321 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     1381 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
     1441 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     1501 gccaagtccg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
     1561 gtacatgacc ttacgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     1621 taccatggtg atgcggtttt ggcagtacac caatgggcgt ggatagcggt ttgactcacg
     1681 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
     1741 acgggacttt ccaaaatgtc gtaataaccc cgccccgttg acgcaaatgg gcggtaggcg
     1801 tgtacggtgg gaggtcaaat cgaattctaa tactgttgac aattaatcat cggcatagta
     1861 tatcggcata gtataatacg acaaggtgag gaactaaacc caggaggcag atcatggaga
     1921 aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa gaacattttg
     1981 aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg gatattacgg
     2041 cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt attcacattc
     2101 ttgcccgcct gatgaatgct catccggaat tccgtatggc aatgaaagac ggtgagctgg
     2161 tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact gaaacgtttt
     2221 catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata tattcgcaag
     2281 atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt gagaatatgt
     2341 ttttcgtctc agccaatccc tgggtgagtt tcaccagttt tgatttaaac gtggccaata
     2401 tggacaactt cttcgccccc gttttcacta tgggcaaata ttatacgcaa ggcgacaagg
     2461 tgctgatgcc gctggcgatt caggttcatc atgccgtctg tgatggcttc catgtcggca
     2521 gaatgcttaa tgaattacaa cagtactgcg atgagtggca gggcggggcg taagcgggac
     2581 tctggggttc gaaatgaccg accaagcgac gcccaaccgg tatcagctca ctcaaaggcg
     2641 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     2701 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2761 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2821 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2881 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2941 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3001 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3061 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3121 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3181 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3241 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     3301 cagcagatta cgcgcagaaa aaaaggattt caagaagatc ctttgatctt ttctacgggg
     3361 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     3421 aggatcttca cctagatcct tttatagtcc ggaatcgtca cacaaaaaac caacacacag
     3481 atgtaatgaa aataaagata ttttattgcg gccatcgtga tggctagagt tcagaagaac
     3541 tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc
     3601 acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac
     3661 gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag
     3721 cggccatttt ccaccatgat attcggcaag caggcatcgc cgtgggtcac gacgagatcc
     3781 tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga
     3841 tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc
     3901 tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc
     3961 cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg
     4021 agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg
     4081 tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg
     4141 tcctgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc
     4201 tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca
     4261 tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca
     4321 atcatggtgg catcgattcc cggtcaacat ccagtacacc tccggcacac cggcttcccc
     4381 aagggcgcca cgctctccca ccacaacatc ctcaacaacg gctactgggt gggccgcacg
     4441 gtcggctaca cggagcagga ccgggtgtgt ctgccggtgc ccttctacca ctgcttcggc
     4501 atggtcatgg gcaacttggg cgccacctcc cacgccgcct gcatcgtcat cccggccccg
     4561 tccttcgagc cggcggcgac gctggaggcg gtgcagcggg agaggtgcac gtccctgtac
     4621 ggcgtcccca cgaagccgaa ttccaccaca ctggactaga tgcatgctcg acagtgtgat
     4681 ggatatctgc agaattcggc tttcacgaca cctgaaatgg aagaaaaaaa ctttgaacca
     4741 ctgtctgagg cttgagaatg aaccaagatc caaactcaaa aagggcaaat tccaaggaga
     4801 attacatcaa gtgccaagct ggcctaactt cagtctccac ccactcagtg tggggaaact
     4861 ccatcgcata aaacccctcc ccccaaccta aagacgacgt actccaaaag ctcgagaact
     4921 aatcgaggtg cctggacggc gcccggtact ccgtggagtc acatgaagcg acggctgagg
     4981 acggaaaggc ccttttcctt tgtgtgggtg actcacccgc ccgctctccc gagcgccgcg
     5041 tcctccattt tgagctgcct gcagcagggc cgggaagcgg ccatctttcc gctcacgcaa
     5101 ctggtgccga ccgggccagc cttgccgccc agggcggggc gatacacggc ggcgcgaggc
     5161 caggcaccag agcaggccgg ccagcttgag actacccccg tccgattctc ggtggccgcg
     5221 ctcgcaggcc ccgcctcgcc gaacatgtgc gctgggacgc acgggccccg tcgccgcccg
     5281 cggccccaaa aaccgaaata ccagtgtgca gatcttggcc cgcatttaca agactatctt
     5341 gccagaaaaa aagcgtcgca gcaggtcatc aaaaatttta aatggctaga gacttatcga
     5401 aagcagcgag acaggcgcga aggtgccacc agattcgcac gcggcggccc cagcgcccag
     5461 gccaggcctc aactcaagca cgaggcgaag gggctcctta agcgcaaggc ctcgaactct
     5521 cccacccact tccaacccga agctcgggat caagaatcac gtactgcagc caggtggaag
     5581 taattcaagg cacgcaaggg ccataacccg taaagaggcc aggcccgcgg gaaccacaca
     5641 cggcacttac ctgtgttctg gcggcaaacc cgttgcgaaa aagaacgttc acggcgacta
     5701 ctgcacttat atacggttct cccccaccct cgggaaaaag gcggagccag tacacgacat
     5761 cactttccca gtttaccccg cgccaccttc tctaggcacc ggttcaattg ccgacccctc
     5821 cccccaactt ctcggggact gtgggcgatg tgcgctctgc ccactgacgg gcaccggagc
     5881 c
//
