LOCUS       sequence-474675-        6768 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express NanoLuc(R)-TAZ 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 6768)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6768)
  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            "Unpublished"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-TAZ Vector
FEATURES             Location/Qualifiers
     source          1..6768
                     /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(1522..2034)
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     promoter        complement(2073..2091)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2074..2091)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        2211..2514
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2669..2689
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        2722..2769
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2801..3460
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2867..2886)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      complement(3632..4220)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3712..3731)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4346..4394
                     /note="synthetic polyadenylation signal"
     CDS             complement(4418..5212)
                     /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(4529..4548)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5139..5158
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5590..6768)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          5599..6537
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(5622..5642)
                     /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 ctagaggatc cccgggtacc gagcccgaat tcgtttaaac
      301 cagccaggtt agaaagggct cacttttgtt cagagcagac tctacatcat tgaagagggg
      361 gatcaggtct tcagattcca aagttcctaa gtcaacgttt gttcctggaa gacagtcaag
      421 gaaatcaggg aaacgggtct gttggggatt gatgttcatg ggtgtttgtc ctgcgttttc
      481 tcctgtatcc atctcatcca cattgctgag gaagtcctcc ggagttgtgg ggacactgta
      541 gcaccctaac cccaggccac tgtcagtgct ctgctccctc gaatgatatg gccctccatt
      601 gaggaaagga tctgagctat tattagtgat ggatctcatg tctggggtca tcgtgggtgg
      661 gttgacagca gcctgaactg gggcaagagt ctcagcttcc atggggagct gtcgacagag
      721 ggcagcttcc tgcctcatga gctcctcttg gcgcattcga atcctttctc tctccatctg
      781 gattctctga agccgcagtt tctgctgctg ctgctgctga gtggtcagcg cattgggcat
      841 actcatgagc cctgcgggtg ggttctgagt ggggtggttc tgctggctca gggtactggg
      901 ggccatctgc tgctggtgtt ggtgattcat cacgagattt ggctgggata ctgccatgga
      961 cctctgaggc actggtgtgg aactgacggc agggtggagg ttcatatgat tcagaggctg
     1021 attcatcgcc ttcctagggt cttgccatgt ggtgattttt tctatgtgat tgaggaagta
     1081 cctctggcca gtggccgtga aggtcatctc ccagcccggg ggcagtggca gctcgtcggt
     1141 cacgtcgtag gactgctggc ggaggtgcgc gtgctgctgc gcggggctac ccgcagcacc
     1201 cgcgccggtg cccagctgca gggacgcggg cgacgagtgc gagcggacat gctgggcacc
     1261 cccagccagt cgaggccccg ggtggccgcc cgacgagtcg gtgctggact ggcgcgagtg
     1321 cgagcccgaa tcaggctcct taaagaaaga ctccggcagg atcttcttcc gccacgagct
     1381 aggcttcgga ttcatgacag agttgaagag ggcttcgagg tctgtgtcta ggtcctgcgt
     1441 gacgtggatc acttgctgcc caggcggcgg gagcggaggg ggcgccgagg ccggattcat
     1501 ggcgatcgcg ccgctcgagc ccgccagaat gcgttcgcac agccgccagc cggtcactcc
     1561 gttgatggtt actcggaaca gcagggagcc gtcggggttg atcaggcgct cgtcgataat
     1621 tttgttgccg ttccacaggg tccctgttac agtgatcttt ttgccgtcga acacggcgat
     1681 gccttcatac ggccgtccga aatagtcgat catgttcggc gtaaccccgt cgattaccag
     1741 tgtgccatag tgcaggatca ccttaaagtg atgatcatcc acagggtaca ccaccttaaa
     1801 aattttttcg atctggccca tttggtcgcc gctcagacct tcatacggga tgatgacatg
     1861 gatgtcgatc ttcagcccat tttcaccgct caggacaatc ctttggatcg gagttacgga
     1921 caccccgaga ttctgaaaca aactggacac acctccctgt tcaaggactt ggtccaggtt
     1981 gtagccggct gtctgtcgcc agtccccaac gaaatcttcg agtgtgaaga ccatggtgag
     2041 cgctagctgc aaaaagaaca agtagcttgt attctatagt gtcacctaaa tgggcgatct
     2101 gacggttcac taaaccagct ctgcttatat aggcctccct ggcgcgccac tagttattaa
     2161 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     2221 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     2281 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     2341 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtccgccc
     2401 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     2461 cgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
     2521 cggttttggc agtacaccaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     2581 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     2641 aaatgtcgta ataaccccgc cccgttgacg caaatgggcg gtaggcgtgt acggtgggag
     2701 gtcaaatcga attctaatac tgttgacaat taatcatcgg catagtatat cggcatagta
     2761 taatacgaca aggtgaggaa ctaaacccag gaggcagatc atggagaaaa aaatcactgg
     2821 atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc
     2881 agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac
     2941 cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat
     3001 gaatgctcat ccggaattcc gtatggcaat gaaagacggt gagctggtga tatgggatag
     3061 tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag
     3121 tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta
     3181 cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc
     3241 caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt
     3301 cgcccccgtt ttcactatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct
     3361 ggcgattcag gttcatcatg ccgtctgtga tggcttccat gtcggcagaa tgcttaatga
     3421 attacaacag tactgcgatg agtggcaggg cggggcgtaa gcgggactct ggggttcgaa
     3481 atgaccgacc aagcgacgcc caaccggtat cagctcactc aaaggcggta atacggttat
     3541 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3601 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3661 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3721 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3781 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3841 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3901 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3961 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4021 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     4081 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4141 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4201 gcagaaaaaa aggatttcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4261 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4321 agatcctttt atagtccgga atcgtcacac aaaaaaccaa cacacagatg taatgaaaat
     4381 aaagatattt tattgcggcc atcgtgatgg ctagagttca gaagaactcg tcaagaaggc
     4441 gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt
     4501 cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat
     4561 agcggtccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca
     4621 ccatgatatt cggcaagcag gcatcgccgt gggtcacgac gagatcctcg ccgtcgggca
     4681 tgcgcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca
     4741 gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt
     4801 tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat
     4861 cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga tcctgccccg
     4921 gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agcacagctg
     4981 cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtcc tgcagttcat
     5041 tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc
     5101 ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc
     5161 tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc atggtggcat
     5221 cgattcccgg tcaacatcca gtacacctcc ggcacaccgg cttccccaag ggcgccacgc
     5281 tctcccacca caacatcctc aacaacggct actgggtggg ccgcacggtc ggctacacgg
     5341 agcaggaccg ggtgtgtctg ccggtgccct tctaccactg cttcggcatg gtcatgggca
     5401 acttgggcgc cacctcccac gccgcctgca tcgtcatccc ggccccgtcc ttcgagccgg
     5461 cggcgacgct ggaggcggtg cagcgggaga ggtgcacgtc cctgtacggc gtccccacga
     5521 agccgaattc caccacactg gactagatgc atgctcgaca gtgtgatgga tatctgcaga
     5581 attcggcttt cacgacacct gaaatggaag aaaaaaactt tgaaccactg tctgaggctt
     5641 gagaatgaac caagatccaa actcaaaaag ggcaaattcc aaggagaatt acatcaagtg
     5701 ccaagctggc ctaacttcag tctccaccca ctcagtgtgg ggaaactcca tcgcataaaa
     5761 cccctccccc caacctaaag acgacgtact ccaaaagctc gagaactaat cgaggtgcct
     5821 ggacggcgcc cggtactccg tggagtcaca tgaagcgacg gctgaggacg gaaaggccct
     5881 tttcctttgt gtgggtgact cacccgcccg ctctcccgag cgccgcgtcc tccattttga
     5941 gctgcctgca gcagggccgg gaagcggcca tctttccgct cacgcaactg gtgccgaccg
     6001 ggccagcctt gccgcccagg gcggggcgat acacggcggc gcgaggccag gcaccagagc
     6061 aggccggcca gcttgagact acccccgtcc gattctcggt ggccgcgctc gcaggccccg
     6121 cctcgccgaa catgtgcgct gggacgcacg ggccccgtcg ccgcccgcgg ccccaaaaac
     6181 cgaaatacca gtgtgcagat cttggcccgc atttacaaga ctatcttgcc agaaaaaaag
     6241 cgtcgcagca ggtcatcaaa aattttaaat ggctagagac ttatcgaaag cagcgagaca
     6301 ggcgcgaagg tgccaccaga ttcgcacgcg gcggccccag cgcccaggcc aggcctcaac
     6361 tcaagcacga ggcgaagggg ctccttaagc gcaaggcctc gaactctccc acccacttcc
     6421 aacccgaagc tcgggatcaa gaatcacgta ctgcagccag gtggaagtaa ttcaaggcac
     6481 gcaagggcca taacccgtaa agaggccagg cccgcgggaa ccacacacgg cacttacctg
     6541 tgttctggcg gcaaacccgt tgcgaaaaag aacgttcacg gcgactactg cacttatata
     6601 cggttctccc ccaccctcgg gaaaaaggcg gagccagtac acgacatcac tttcccagtt
     6661 taccccgcgc caccttctct aggcaccggt tcaattgccg acccctcccc ccaacttctc
     6721 ggggactgtg ggcgatgtgc gctctgccca ctgacgggca ccggagcc
//
