LOCUS       sequence-476793-        5237 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express NanoLuc(R)-TAF1(BD2) 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 5237)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5237)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: NanoLuc-TAF1(BD2) Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5237
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(19..37)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    74..195
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(85..104)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     139..158
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        440..797
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        440..631
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(463..483)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      648..783
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        812..859
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             891..1685
                     /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(945..964)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1555..1574
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1749..1797
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(2036..2624)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2116..2135)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2740..3023
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     enhancer        3214..3593
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3594..3797
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3748..3768
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          3933..4065
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     4048..4067
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     4109..4128
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4109..4127
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4141..4653
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
ORIGIN
        1 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcggccgctt
       61 cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
      121 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
      181 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
      241 atgtgggagg tttttttaag caagtaaaac ctctacaaat gtggtaaaat cgaattttaa
      301 caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt
      361 atttcacacc gcatacgcgg atctgcgcag caccatggcc tgaaataacc tctgaaagag
      421 gaacttggtt aggtaccttc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
      481 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
      541 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
      601 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
      661 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
      721 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
      781 gcctaggctt ttgcaaaaag cttaattaac tgttgacaat taatcatcgg catagtatat
      841 cggcatagta taatacgaca aggtgaggaa ctaaacccag gaggcagatc atgattgaac
      901 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
      961 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     1021 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     1081 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     1141 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     1201 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     1261 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     1321 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     1381 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     1441 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     1501 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     1561 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     1621 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     1681 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     1741 atggccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgaat
     1801 cgatagcgat aaggatcctc tttgcgcttg cgttttccct tgtccagata gcccagtagc
     1861 tgacattcat ccggggtcag caccgtttct gcggactggc tttctacccg gtatcagctc
     1921 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     1981 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     2041 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2101 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2161 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2221 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2281 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2341 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2401 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2461 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     2521 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     2581 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatt tcaagaagat cctttgatct
     2641 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     2701 gattatcaaa aaggatcttc acctagatcc ttttatagtc cggaaataca ggaacgcacg
     2761 ctggatggcc cttcgctggg atggtgaaac catgaaaaat ggcagcttca gtggattaag
     2821 tgggggtaat gtggcctgta ccctctggtt gcataggtat tcatacggtt aaaatttatc
     2881 aggcgcgatt gcggcagttt ttcgggtggt ttgttgccat ttttacctgt ctgctgccgt
     2941 gatcgcgctg aacgcgtttt agcggtgcgt acaattaagg gattatggta aatccactta
     3001 ctgtctgccc tcgtagccat cgagataaac cgcagtactc cggccacgat gcgtccggcg
     3061 tagaggatcg agatcttcaa tattggccat tagccatatt attcattggt tatatagcat
     3121 aaatcaatat tggctattgg ccattgcata cgttgtatct atatcataat atgtacattt
     3181 atattggctc atgtccaata tgaccgccat gttggcattg attattgact agttattaat
     3241 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     3301 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     3361 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     3421 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtccgcccc
     3481 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttac
     3541 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
     3601 ggttttggca gtacaccaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     3661 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     3721 aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg taggcgtgta cggtgggagg
     3781 tctatataag cagagctggt ttagtgaacc gtcagatcac tagaagcttt attgcggtag
     3841 tttatcacag ttaaattgct aacgcagtca gtgcttctga cacaacagtc tcgaacttaa
     3901 gctgcagaag ttggtcgtga ggcactgggc aggtaagtat caaggttaca agacaggttt
     3961 aaggagacca atagaaactg ggcttgtcga gacagagaag actcttgcgt ttctgatagg
     4021 cacctattgg tcttactgac atccactttg cctttctctc cacaggtgtc cactcccagt
     4081 tcaattacag ctcttaaggc tagagtatta atacgactca ctatagggct agcgctcacc
     4141 atggtcttca cactcgaaga tttcgttggg gactggcgac agacagccgg ctacaacctg
     4201 gaccaagtcc ttgaacaggg aggtgtgtcc agtttgtttc agaatctcgg ggtgtccgta
     4261 actccgatcc aaaggattgt cctgagcggt gaaaatgggc tgaagatcga catccatgtc
     4321 atcatcccgt atgaaggtct gagcggcgac caaatgggcc agatcgaaaa aatttttaag
     4381 gtggtgtacc ctgtggatga tcatcacttt aaggtgatcc tgcactatgg cacactggta
     4441 atcgacgggg ttacgccgaa catgatcgac tatttcggac ggccgtatga aggcatcgcc
     4501 gtgttcgacg gcaaaaagat cactgtaaca gggaccctgt ggaacggcaa caaaattatc
     4561 gacgagcgcc tgatcaaccc cgacggctcc ctgctgttcc gagtaaccat caacggagtg
     4621 accggctggc ggctgtgcga acgcattctg gcgggctcga gcggcgcgat cgccatggct
     4681 atcaacccct tgctggatga tgatgaccaa gtggcgtttt ctttcattct ggacaacatt
     4741 gtcacccaga aaatgatggc agttccagat tcttggccat ttcatcaccc agttaataag
     4801 aaatttgttc cagattatta caaagtgatt gtcaatccaa tggatttaga gaccatacgt
     4861 aagaacatct ccaagcacaa gtatcagagt cgggagagct ttctggatga tgtaaacctt
     4921 attctggcca acagtgttaa gtataatgga cctgagagtc agtatactaa gactgcccag
     4981 gagattgtga acgtctgtta ccagacattg actgagtatg atgaacattt gactcaactt
     5041 gagaaggata tttgtactgc taaagaagca gctttggagg aagcagaatt agaaagcctg
     5101 gacccaatga ccccagggcc ctacacgcct cagcctcctg atgtttaaac gaattcgggc
     5161 tcggtacccg gggatcctct agagtcgacc tgcaggcatg caagctgatc cggctgctaa
     5221 caaagcccga aaggaag
//
