LOCUS       sequence-470732-       10359 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express TAF1-NanoLuc(R) 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 10359)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10359)
  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            "Unpublished"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: NanoBRET Assay Vector, TAF1-NanoLuc
FEATURES             Location/Qualifiers
     source          1..10359
                     /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             complement(2575..2589)
                     /codon_start=1
                     /product="tag for protein cleavage using Ni2+ ion (Dang et 
                     al., 2019)"
                     /label=SNAC tag
                     /translation="GSHHW"
     CDS             6702..7214
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(7302..7320)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    7357..7478
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7368..7387)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7422..7441
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        7723..8080
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        7723..7914
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(7746..7766)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      7931..8066
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        8095..8142
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             8174..8968
                     /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(8228..8247)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8838..8857
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    9032..9080
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(9319..9907)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9399..9418)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    10023..10306
                     /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 cgccatggga cccggctgcg
     1081 atttgctgct gcggacagca gctaccatca ctgctgccgc catcatgtca gacacggaca
     1141 gcgacgaaga ttccgctgga ggcggcccat tttctttagc gggtttcctt ttcggcaaca
     1201 tcaatggagc cgggcagctg gagggggaaa gcgtcttgga tgatgaatgt aagaagcact
     1261 tggcaggctt gggggctttg gggctgggca gcctgatcac tgaactcacg gcaaatgaag
     1321 aattgaccgg gactgacggt gccttggtaa atgatgaagg gtgggttagg agtacagaag
     1381 atgctgtgga ctattcagac atcaatgagg tggcagaaga tgaaagccga agataccagc
     1441 agacgatggg gagcttgcag cccctttgcc actcagatta tgatgaagat gactatgatg
     1501 ctgattgtga agacattgat tgcaagttga tgcctcctcc acctccaccc ccgggaccaa
     1561 tgaagaagga taaggaccag gattctatta ctggtgagaa agtggacttc agtagttcct
     1621 ctgactcaga atctgagatg ggacctcagg aagcaacaca ggcagaatct gaagatggaa
     1681 agctgaccct tccattggct gggattatgc agcatgatgc caccaagctg ttgccaagtg
     1741 tcacagaact ttttccagaa tttcgacctg gaaaggtgtt acgttttcta cgtctttttg
     1801 gaccagggaa gaatgtccca tctgtttggc ggagtgctcg gagaaagagg aagaagaagc
     1861 accgtgagct gatacaggaa gagcagatcc aggaggtgga gtgctcagta gaatcagaag
     1921 tcagccagaa gtctttgtgg aactacgact acgctccacc accacctcca gagcagtgtc
     1981 tctctgatga tgaaatcacg atgatggctc ctgtggagtc caaattttcc caatcaactg
     2041 gagatataga taaagtgaca gataccaaac caagagtggc tgagtggcgt tatgggcctg
     2101 cccgactgtg gtatgatatg ctgggtgtcc ctgaagatgg cagtgggttt gactatggct
     2161 tcaaactgag aaagacagaa catgaacctg tgataaaatc tagaatgata gaggaattta
     2221 ggaaacttga ggaaaacaat ggcactgatc ttctggctga tgaaaacttc ctgatggtga
     2281 cacagctgca ttgggaggat gatatcatct gggatgggga ggatgtcaaa cacaaaggga
     2341 caaaacctca gcgtgcaagc ctggcaggct ggcttccttc tagcatgact aggaatgcga
     2401 tggcttacaa tgttcagcaa ggttttgcag ccactcttga tgatgacaaa ccttggtact
     2461 ccatttttcc cattgacaat gaggatctgg tatatggacg ctgggaggac aatatcattt
     2521 gggatgctca ggccatgccc cggctgttgg aacctcctgt tttgacactt gatcccaatg
     2581 atgagaacct cattttggaa attcctgatg agaaggaaga ggccacctct aactccccct
     2641 ccaaggagag taagaaggaa tcatctctga agaagagtcg aattctctta gggaaaacag
     2701 gagtcatcaa ggaggaacca cagcagaaca tgtctcagcc agaagtgaaa gatccatgga
     2761 atctctccaa tgatgagtat tattatccca agcaacaggg tcttcgaggc acctttggag
     2821 ggaatattat ccagcattca attcctgctg tggaattacg gcagcccttc tttcccaccc
     2881 acatggggcc catcaaactc cggcagttcc atcgcccacc tctgaaaaag tactcatttg
     2941 gtgcactttc tcagccaggt ccccactcag tccaaccttt gctaaagcac atcaaaaaaa
     3001 aggccaagat gagagaacaa gagaggcaag cttcaggtgg tggagagatg ttttttatgc
     3061 gcacacctca ggacctcaca ggcaaagatg gtgatcttat tcttgcagaa tatagtgagg
     3121 aaaatggacc cttaatgatg caggttggca tggcaaccaa gataaagaac tattataaac
     3181 ggaaacctgg aaaagatcct ggagcaccag attgtaaata tggggaaact gtttactgcc
     3241 atacatctcc tttcctgggt tctctccatc ctggccaatt gctgcaagca tttgagaaca
     3301 acctttttcg tgctccaatt tatcttcata agatgccaga aactgatttc ttgatcattc
     3361 ggacaagaca gggttactat attcgggaat tagtggatat ttttgtggtt ggccagcagt
     3421 gtcccttgtt tgaagttcct gggcctaact ccaaaagggc caatacgcat attcgagact
     3481 ttctacaggt ttttatttac cgccttttct ggaaaagtaa agatcggcca cggaggatac
     3541 gaatggaaga tataaaaaaa gcctttcctt cccattcaga aagcagcatc cggaagaggc
     3601 taaagctctg cgctgacttc aaacgcacag ggatggactc aaactggtgg gtgcttaagt
     3661 ctgattttcg tttaccaacg gaagaagaga tcagagctat ggtgtcacca gagcagtgct
     3721 gtgcttatta tagcatgata gctgcagagc aacgactgaa ggatgctggc tatggtgaga
     3781 aatccttttt tgctccagaa gaagaaaatg aggaagattt ccagatgaag attgatgatg
     3841 aagttcgcac tgccccttgg aacaccacaa gggccttcat tgctgccatg aagggcaagt
     3901 gtctgctaga ggtgactggg gtggcagatc ccacggggtg tggtgaagga ttctcctatg
     3961 tgaagattcc aaacaaacca acacagcaga aggatgataa agaaccgcag ccagtgaaga
     4021 agacagtgac aggaacagat gcagaccttc gtcgcctttc cctgaaaaat gccaagcaac
     4081 ttctacgtaa atttggtgtg cctgaggaag agattaaaaa gttgtcccgc tgggaagtga
     4141 ttgatgtggt gcgcacaatg tcaacagaac aggctcgttc tggagagggg cccatgagta
     4201 aatttgcccg tggatcaagg ttttctgtgg ctgagcatca agagcgttac aaagaggaat
     4261 gtcagcgcat ctttgaccta cagaacaagg ttctgtcatc aactgaagtc ttatcaactg
     4321 acacagacag cagctcagct gaagatagtg actttgaaga aatgggaaag aacattgaga
     4381 acatgttgca gaacaagaaa accagctctc agctttcacg tgaacgggag gaacaggagc
     4441 ggaaggaact acagcgaatg ctactggcag caggctcagc agcatccgga aacaatcaca
     4501 gagatgatga cacagcttcc gtgactagcc ttaactcttc tgccactgga cgctgtctca
     4561 agatttatcg cacgtttcga gatgaagagg ggaaagagta tgttcgctgt gagacagtcc
     4621 gaaaaccagc tgtcattgat gcctatgtgc gcatacggac tacaaaagat gaggaattca
     4681 ttcgaaaatt tgcccttttt gatgaacaac atcgggaaga gatgcgaaaa gaacggcgga
     4741 ggattcaaga gcaactgagg cggcttaaga ggaaccagga aaaggagaag cttaagggtc
     4801 ctcctgagaa gaagcccaag aaaatgaagg agcgtcctga cctaaaactg aaatgtgggg
     4861 catgtggtgc cattggacac atgaggacta acaaattctg ccccctctat tatcaaacaa
     4921 atgcgccacc ttccaaccct gttgccatga cagaagaaca ggaggaggag ttggaaaaga
     4981 cagtcattca taatgataat gaagaactta tcaaggttga agggaccaaa attgtcttgg
     5041 ggaaacagct aattgagagt gcggatgagg ttcgcagaaa atctctggtt ctcaagtttc
     5101 ctaaacagca gcttcctcca aagaagaaac ggcgagttgg aaccactgtt cactgtgact
     5161 atttgaatag acctcataag tccatccacc ggcgccgcac agaccctatg gtgacgctgt
     5221 cgtccatctt ggagtctatc atcaatgaca tgagagatct tccaaataca taccctttcc
     5281 acactccagt caatgcaaag gttgtaaagg actactacaa aatcatcact cggccaatgg
     5341 acctacaaac actccgcgaa aacgtgcgta aacgcctcta cccatctcgg gaagagttca
     5401 gagagcatct ggagctaatt gtgaaaaata gtgcaaccta caatgggcca aaacactcat
     5461 tgactcagat ctctcaatcc atgctggatc tctgtgatga aaaactcaaa gagaaagaag
     5521 acaaattagc tcgcttagag aaagctatca accccttgct ggatgatgat gaccaagtgg
     5581 cgttttcttt cattctggac aacattgtca cccagaaaat gatggcagtt ccagattctt
     5641 ggccatttca tcacccagtt aataagaaat ttgttccaga ttattacaaa gtgattgtca
     5701 atccaatgga tttagagacc atacgtaaga acatctccaa gcacaagtat cagagtcggg
     5761 agagctttct ggatgatgta aaccttattc tggccaacag tgttaagtat aatggacctg
     5821 agagtcagta tactaagact gcccaggaga ttgtgaacgt ctgttaccag acattgactg
     5881 agtatgatga acatttgact caacttgaga aggatatttg tactgctaaa gaagcagctt
     5941 tggaggaagc agaattagaa agcctggacc caatgacccc agggccctac acgcctcagc
     6001 ctcctgattt gtatgatacc aacacatccc tcagtatgtc tcgagatgcc tctgtatttc
     6061 aagatgagag caatatgtct gtcttggata ttcccagtgc cactccagaa aagcaggtaa
     6121 cacaggaagg tgaagatgga gatggtgatc ttgcagatga agaggaagga actgtacaac
     6181 agcctcaagc cagtgtcctg tatgaggatt tgcttatgtc tgaaggagaa gatgatgagg
     6241 aagatgctgg gagtgatgaa gaaggagaca atcctttctc tgctatccag ctgagtgaaa
     6301 gtggaagtga ctctgatgtg ggatctggtg gaataagacc caaacaaccc cgcatgcttc
     6361 aggagaacac aaggatggac atggaaaatg aagaaagcat gatgtcctat gagggagacg
     6421 gtggggaggc ttcccatggt ttggaggata gcaacatcag ttatgggagc tatgaggagc
     6481 ctgatcccaa gtcgaacacc caagacacaa gcttcagcag catcggtggg tatgaggtat
     6541 cagaggagga agaagatgag gaggaggaag agcagcgctc tgggccgagc gtactaagcc
     6601 aggtccacct gtcagaggac gaggaggaca gtgaggattt ccactccatt gctggggaca
     6661 gtgacttgga ctctgatgaa gtttctctcg gctcgagcgg cgtcttcaca ctcgaagatt
     6721 tcgttgggga ctggcgacag acagccggct acaacctgga ccaagtcctt gaacagggag
     6781 gtgtgtccag tttgtttcag aatctcgggg tgtccgtaac tccgatccaa aggattgtcc
     6841 tgagcggtga aaatgggctg aagatcgaca tccatgtcat catcccgtat gaaggtctga
     6901 gcggcgacca aatgggccag atcgaaaaaa tttttaaggt ggtgtaccct gtggatgatc
     6961 atcactttaa ggtgatcctg cactatggca cactggtaat cgacggggtt acgccgaaca
     7021 tgatcgacta tttcggacgg ccgtatgaag gcatcgccgt gttcgacggc aaaaagatca
     7081 ctgtaacagg gaccctgtgg aacggcaaca aaattatcga cgagcgcctg atcaaccccg
     7141 acggctccct gctgttccga gtaaccatca acggagtgac cggctggcgg ctgtgcgaac
     7201 gcattctggc gtaaggccgc gactctagag tcgacctgca ggcatgcaag ctgatccggc
     7261 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
     7321 ataacccctt ggggcggccg cttcgagcag acatgataag atacattgat gagtttggac
     7381 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     7441 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     7501 ttatgtttca ggttcagggg gagatgtggg aggttttttt aagcaagtaa aacctctaca
     7561 aatgtggtaa aatcgaattt taacaaaata ttaacgctta caatttcctg atgcggtatt
     7621 ttctccttac gcatctgtgc ggtatttcac accgcatacg cggatctgcg cagcaccatg
     7681 gcctgaaata acctctgaaa gaggaacttg gttaggtacc ttctgaggcg gaaagaacca
     7741 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     7801 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     7861 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     7921 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     7981 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     8041 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagcttaatt aactgttgac
     8101 aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc
     8161 caggaggcag atcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     8221 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     8281 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     8341 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     8401 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     8461 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     8521 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     8581 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     8641 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     8701 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     8761 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     8821 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     8881 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     8941 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     9001 gcgacgccca acctgccatc acgatggccg caataaaata tctttatttt cattacatct
     9061 gtgtgttggt tttttgtgtg aatcgatagc gataaggatc ctctttgcgc ttgcgttttc
     9121 ccttgtccag atagcccagt agctgacatt catccggggt cagcaccgtt tctgcggact
     9181 ggctttctac ccggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     9241 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     9301 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     9361 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     9421 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     9481 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     9541 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     9601 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     9661 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     9721 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     9781 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     9841 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     9901 atttcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     9961 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttata
    10021 gtccggaaat acaggaacgc acgctggatg gcccttcgct gggatggtga aaccatgaaa
    10081 aatggcagct tcagtggatt aagtgggggt aatgtggcct gtaccctctg gttgcatagg
    10141 tattcatacg gttaaaattt atcaggcgcg attgcggcag tttttcgggt ggtttgttgc
    10201 catttttacc tgtctgctgc cgtgatcgcg ctgaacgcgt tttagcggtg cgtacaatta
    10261 agggattatg gtaaatccac ttactgtctg ccctcgtagc catcgagata aaccgcagta
    10321 ctccggccac gatgcgtccg gcgtagagga tcgagatct
//
