LOCUS       sequence-474724-        8234 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express NanoLuc(R)-TAF1 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 8234)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8234)
  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            "Unpublished"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 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..8234
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        101..480
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        481..684
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     635..655
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          820..952
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     935..954
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     996..1015
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        996..1014
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1028..1540
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     primer_bind     complement(5140..5158)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    5195..5316
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5206..5225)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5260..5279
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5561..5918
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5561..5752
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5584..5604)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5769..5904
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5933..5980
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6012..6806
                     /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(6066..6085)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6676..6695
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6870..6918
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(7157..7745)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7237..7256)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7861..8144
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 atagcataaa tcaatattgg ctattggcca ttgcatacgt tgtatctata tcataatatg
       61 tacatttata ttggctcatg tccaatatga ccgccatgtt ggcattgatt attgactagt
      121 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
      181 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
      241 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      301 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      361 ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
      421 accttacggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      481 gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt
      541 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      601 tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg
      661 tgggaggtct atataagcag agctggttta gtgaaccgtc agatcactag aagctttatt
      721 gcggtagttt atcacagtta aattgctaac gcagtcagtg cttctgacac aacagtctcg
      781 aacttaagct gcagaagttg gtcgtgaggc actgggcagg taagtatcaa ggttacaaga
      841 caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact cttgcgtttc
      901 tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac aggtgtccac
      961 tcccagttca attacagctc ttaaggctag agtattaata cgactcacta tagggctagc
     1021 gctcaccatg gtcttcacac tcgaagattt cgttggggac tggcgacaga cagccggcta
     1081 caacctggac caagtccttg aacagggagg tgtgtccagt ttgtttcaga atctcggggt
     1141 gtccgtaact ccgatccaaa ggattgtcct gagcggtgaa aatgggctga agatcgacat
     1201 ccatgtcatc atcccgtatg aaggtctgag cggcgaccaa atgggccaga tcgaaaaaat
     1261 ttttaaggtg gtgtaccctg tggatgatca tcactttaag gtgatcctgc actatggcac
     1321 actggtaatc gacggggtta cgccgaacat gatcgactat ttcggacggc cgtatgaagg
     1381 catcgccgtg ttcgacggca aaaagatcac tgtaacaggg accctgtgga acggcaacaa
     1441 aattatcgac gagcgcctga tcaaccccga cggctccctg ctgttccgag taaccatcaa
     1501 cggagtgacc ggctggcggc tgtgcgaacg cattctggcg ggctcgagcg gcgcgatcgc
     1561 catgggaccc ggctgcgatt tgctgctgcg gacagcagct accatcactg ctgccgccat
     1621 catgtcagac acggacagcg acgaagattc cgctggaggc ggcccatttt ctttagcggg
     1681 tttccttttc ggcaacatca atggagccgg gcagctggag ggggaaagcg tcttggatga
     1741 tgaatgtaag aagcacttgg caggcttggg ggctttgggg ctgggcagcc tgatcactga
     1801 actcacggca aatgaagaat tgaccgggac tgacggtgcc ttggtaaatg atgaagggtg
     1861 ggttaggagt acagaagatg ctgtggacta ttcagacatc aatgaggtgg cagaagatga
     1921 aagccgaaga taccagcaga cgatggggag cttgcagccc ctttgccact cagattatga
     1981 tgaagatgac tatgatgctg attgtgaaga cattgattgc aagttgatgc ctcctccacc
     2041 tccacccccg ggaccaatga agaaggataa ggaccaggat tctattactg gtgagaaagt
     2101 ggacttcagt agttcctctg actcagaatc tgagatggga cctcaggaag caacacaggc
     2161 agaatctgaa gatggaaagc tgacccttcc attggctggg attatgcagc atgatgccac
     2221 caagctgttg ccaagtgtca cagaactttt tccagaattt cgacctggaa aggtgttacg
     2281 ttttctacgt ctttttggac cagggaagaa tgtcccatct gtttggcgga gtgctcggag
     2341 aaagaggaag aagaagcacc gtgagctgat acaggaagag cagatccagg aggtggagtg
     2401 ctcagtagaa tcagaagtca gccagaagtc tttgtggaac tacgactacg ctccaccacc
     2461 acctccagag cagtgtctct ctgatgatga aatcacgatg atggctcctg tggagtccaa
     2521 attttcccaa tcaactggag atatagataa agtgacagat accaaaccaa gagtggctga
     2581 gcatcaagag cgttacaaag aggaatgtca gcgcatcttt gacctacaga acaaggttct
     2641 gtcatcaact gaagtcttat caactgacac agacagcagc tcagctgaag atagtgactt
     2701 tgaagaaatg ggaaagaaca ttgagaacat gttgcagaac aagaaaacca gctctcagct
     2761 ttcacgtgaa cgggaggaac aggagcggaa ggaactacag cgaatgctac tggcagcagg
     2821 ctcagcagca tccggaaaca atcacagaga tgatgacaca gcttccgtga ctagccttaa
     2881 ctcttctgcc actggacgct gtctcaagat ttatcgcacg tttcgagatg aagaggggaa
     2941 agagtatgtt cgctgtgaga cagtccgaaa accagctgtc attgatgcct atgtgcgcat
     3001 acggactaca aaagatgagg aattcattcg aaaatttgcc ctttttgatg aacaacatcg
     3061 ggaagagatg cgaaaagaac ggcggaggat tcaagagcaa ctgaggcggc ttaagaggaa
     3121 ccaggaaaag gagaagctta agggtcctcc tgagaagaag cccaagaaaa tgaaggagcg
     3181 tcctgaccta aaactgaaat gtggggcatg tggtgccatt ggacacatga ggactaacaa
     3241 attctgcccc ctctattatc aaacaaatgc gccaccttcc aaccctgttg ccatgacaga
     3301 agaacaggag gaggagttgg aaaagacagt cattcataat gataatgaag aacttatcaa
     3361 ggttgaaggg accaaaattg tcttggggaa acagctaatt gagagtgcgg atgaggttcg
     3421 cagaaaatct ctggttctca agtttcctaa acagcagctt cctccaaaga agaaacggcg
     3481 agttggaacc actgttcact gtgactattt gaatagacct cataagtcca tccaccggcg
     3541 ccgcacagac cctatggtga cgctgtcgtc catcttggag tctatcatca atgacatgag
     3601 agatcttcca aatacatacc ctttccacac tccagtcaat gcaaaggttg taaaggacta
     3661 ctacaaaatc atcactcggc caatggacct acaaacactc cgcgaaaacg tgcgtaaacg
     3721 cctctaccca tctcgggaag agttcagaga gcatctggag ctaattgtga aaaatagtgc
     3781 aacctacaat gggccaaaac actcattgac tcagatctct caatccatgc tggatctctg
     3841 tgatgaaaaa ctcaaagaga aagaagacaa attagctcgc ttagagaaag ctatcaaccc
     3901 cttgctggat gatgatgacc aagtggcgtt ttctttcatt ctggacaaca ttgtcaccca
     3961 gaaaatgatg gcagttccag attcttggcc atttcatcac ccagttaata agaaatttgt
     4021 tccagattat tacaaagtga ttgtcaatcc aatggattta gagaccatac gtaagaacat
     4081 ctccaagcac aagtatcaga gtcgggagag ctttctggat gatgtaaacc ttattctggc
     4141 caacagtgtt aagtataatg gacctgagag tcagtatact aagactgccc aggagattgt
     4201 gaacgtctgt taccagacat tgactgagta tgatgaacat ttgactcaac ttgagaagga
     4261 tatttgtact gctaaagaag cagctttgga ggaagcagaa ttagaaagcc tggacccaat
     4321 gaccccaggg ccctacacgc ctcagcctcc tgatttgtat gataccaaca catccctcag
     4381 tatgtctcga gatgcctctg tatttcaaga tgagagcaat atgtctgtct tggatattcc
     4441 cagtgccact ccagaaaagc aggtaacaca ggaaggtgaa gatggagatg gtgatcttgc
     4501 agatgaagag gaaggaactg tacaacagcc tcaagccagt gtcctgtatg aggatttgct
     4561 tatgtctgaa ggagaagatg atgaggaaga tgctgggagt gatgaagaag gagacaatcc
     4621 tttctctgct atccagctga gtgaaagtgg aagtgactct gatgtgggat ctggtggaat
     4681 aagacccaaa caaccccgca tgcttcagga gaacacaagg atggacatgg aaaatgaaga
     4741 aagcatgatg tcctatgagg gagacggtgg ggaggcttcc catggtttgg aggatagcaa
     4801 catcagttat gggagctatg aggagcctga tcccaagtcg aacacccaag acacaagctt
     4861 cagcagcatc ggtgggtatg aggtatcaga ggaggaagaa gatgaggagg aggaagagca
     4921 gcgctctggg ccgagcgtac taagccaggt ccacctgtca gaggacgagg aggacagtga
     4981 ggatttccac tccattgctg gggacagtga cttggactct gatgaagttt aaacgaattc
     5041 gggctcggta cccggggatc ctctagagtc gacctgcagg catgcaagct gatccggctg
     5101 ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat
     5161 aaccccttgg ggcggccgct tcgagcagac atgataagat acattgatga gtttggacaa
     5221 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     5281 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     5341 atgtttcagg ttcaggggga gatgtgggag gtttttttaa gcaagtaaaa cctctacaaa
     5401 tgtggtaaaa tcgaatttta acaaaatatt aacgcttaca atttcctgat gcggtatttt
     5461 ctccttacgc atctgtgcgg tatttcacac cgcatacgcg gatctgcgca gcaccatggc
     5521 ctgaaataac ctctgaaaga ggaacttggt taggtacctt ctgaggcgga aagaaccagc
     5581 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     5641 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     5701 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     5761 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     5821 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     5881 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gcttaattaa ctgttgacaa
     5941 ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccca
     6001 ggaggcagat catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     6061 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     6121 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     6181 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     6241 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     6301 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     6361 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     6421 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     6481 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     6541 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     6601 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     6661 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     6721 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     6781 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     6841 gacgcccaac ctgccatcac gatggccgca ataaaatatc tttattttca ttacatctgt
     6901 gtgttggttt tttgtgtgaa tcgatagcga taaggatcct ctttgcgctt gcgttttccc
     6961 ttgtccagat agcccagtag ctgacattca tccggggtca gcaccgtttc tgcggactgg
     7021 ctttctaccc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     7081 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     7141 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     7201 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     7261 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     7321 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     7381 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     7441 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     7501 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     7561 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     7621 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     7681 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     7741 ttcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     7801 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttatagt
     7861 ccggaaatac aggaacgcac gctggatggc ccttcgctgg gatggtgaaa ccatgaaaaa
     7921 tggcagcttc agtggattaa gtgggggtaa tgtggcctgt accctctggt tgcataggta
     7981 ttcatacggt taaaatttat caggcgcgat tgcggcagtt tttcgggtgg tttgttgcca
     8041 tttttacctg tctgctgccg tgatcgcgct gaacgcgttt tagcggtgcg tacaattaag
     8101 ggattatggt aaatccactt actgtctgcc ctcgtagcca tcgagataaa ccgcagtact
     8161 ccggccacga tgcgtccggc gtagaggatc gagatcttca atattggcca ttagccatat
     8221 tattcattgg ttat
//
