LOCUS       sequence-481957-        6870 bp    DNA     circular SYN 04-AUG-2025
DEFINITION  Express NanoLuc-TEAD4 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 6870)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6870)
  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            "Unpublished"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: NanoLuc-TEAD4 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6870
                     /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(1624..2136)
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     promoter        complement(2175..2193)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2176..2193)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        2313..2616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2771..2791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        2824..2871
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2903..3562
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2969..2988)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      complement(3734..4322)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3814..3833)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4448..4496
                     /note="synthetic polyadenylation signal"
     CDS             complement(4520..5314)
                     /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(4631..4650)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5241..5260
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5692..6870)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          5701..6639
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(5724..5744)
                     /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 ttctttcacc agcctgtaga tgtggtgctg agccccgtgc tcactggctg acacctcaaa
      361 gacataggca atgcacagca aggtctcctg tgtgtctctg ttggtgacca cctgcaggat
      421 ggtgaagttc tccagcacgc tgttcatcat gtacttctca gggaggtgct tgagcttgtg
      481 gatgaagttg atcatgtact cacagagcgg ggaccggtgg atgcggtaag agtagtgtcc
      541 attctcatag cgagcatact ctgtctcaac tttctccacc acctgcttgc cgaaagagca
      601 gaccttcgtg gagcaggtga tgatcatgtt ctcggggctc tcatactggc tggagacccc
      661 atagaaggag ctgccttcat cctcgatgtt ggtgttgagg tctgcccaga acttcacaag
      721 aaaaaaggca ttggagggtc cccgttcgaa gagatccttg agtccaccct ttttctccgg
      781 gaatttgtca tagatttggc ggatgtccac ggcttcgagg taggggtcgc tgtagcttgg
      841 gctggactgg ccaatgtgca cgaacaggtg cttgttgtac gtgtccgggt cctgctgctg
      901 ctccaggaag gcagagaact ccaacatcca gagcttggag ctggccacgc tgcggccctg
      961 ccatgggggt gccgggggcg cagagggcga tggggcgggc cctgcaggag actcaaaccc
     1021 tggcagaggc agcggaggct ggacagcata ggtttgctga gagaaaggct tcacatcatg
     1081 ggacgttccg gcttggcctg gcaaagctcc ttgccaaaac cctgagactg ctgggcggcc
     1141 ggggccccgg gcgagggcca tgctactgtg gaaggccgtg gcggagatga tctgtgcaga
     1201 cgacatggca gccatgctct gcagggcctt gtccttagct gcctggtcct ttagcttggc
     1261 ctggatctcg cgagctttgc gacgagccag cacctggatg tggctggaga cctgcttcct
     1321 ggtgcgggtc ttccctgtcc ggagcttgat gtagcgggca atcagctcgt tccgaccata
     1381 catcttgccc tcgtccgaca ggatgatttt gcgcctgcca cagggcgggt agatggcgag
     1441 ggcctcctgg aaactctgct caatatccgg gctccacacg ccctctgcgt cattgtcgat
     1501 gggcttgtcc agtgcctgac tgcccccaga ggcggtgctc ccctcagggg aggtgggaga
     1561 gctccactcg ttggaggtaa tggtgccggc cgtgccctcc atggcgatcg cgccgctcga
     1621 gcccgccaga atgcgttcgc acagccgcca gccggtcact ccgttgatgg ttactcggaa
     1681 cagcagggag ccgtcggggt tgatcaggcg ctcgtcgata attttgttgc cgttccacag
     1741 ggtccctgtt acagtgatct ttttgccgtc gaacacggcg atgccttcat acggccgtcc
     1801 gaaatagtcg atcatgttcg gcgtaacccc gtcgattacc agtgtgccat agtgcaggat
     1861 caccttaaag tgatgatcat ccacagggta caccacctta aaaatttttt cgatctggcc
     1921 catttggtcg ccgctcagac cttcatacgg gatgatgaca tggatgtcga tcttcagccc
     1981 attttcaccg ctcaggacaa tcctttggat cggagttacg gacaccccga gattctgaaa
     2041 caaactggac acacctccct gttcaaggac ttggtccagg ttgtagccgg ctgtctgtcg
     2101 ccagtcccca acgaaatctt cgagtgtgaa gaccatggtg agcgctagct gcaaaaagaa
     2161 caagtagctt gtattctata gtgtcaccta aatgggcgat ctgacggttc actaaaccag
     2221 ctctgcttat ataggcctcc ctggcgcgcc actagttatt aatagtaatc aattacgggg
     2281 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     2341 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     2401 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     2461 cacttggcag tacatcaagt gtatcatatg ccaagtccgc cccctattga cgtcaatgac
     2521 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tacgggactt tcctacttgg
     2581 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacacc
     2641 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     2701 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taataacccc
     2761 gccccgttga cgcaaatggg cggtaggcgt gtacggtggg aggtcaaatc gaattctaat
     2821 actgttgaca attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg
     2881 aactaaaccc aggaggcaga tcatggagaa aaaaatcact ggatatacca ccgttgatat
     2941 atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc aatgtaccta
     3001 taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga aaaataagca
     3061 caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc atccggaatt
     3121 ccgtatggca atgaaagacg gtgagctggt gatatgggat agtgttcacc cttgttacac
     3181 cgttttccat gagcaaactg aaacgttttc atcgctctgg agtgaatacc acgacgattt
     3241 ccggcagttt ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa acctggccta
     3301 tttccctaaa gggtttattg agaatatgtt tttcgtctca gccaatccct gggtgagttt
     3361 caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg ttttcactat
     3421 gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc aggttcatca
     3481 tgccgtctgt gatggcttcc atgtcggcag aatgcttaat gaattacaac agtactgcga
     3541 tgagtggcag ggcggggcgt aagcgggact ctggggttcg aaatgaccga ccaagcgacg
     3601 cccaaccggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     3661 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     3721 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3781 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     3841 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3901 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3961 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4021 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4081 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4141 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     4201 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4261 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatttc
     4321 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4381 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttatagtccg
     4441 gaatcgtcac acaaaaaacc aacacacaga tgtaatgaaa ataaagatat tttattgcgg
     4501 ccatcgtgat ggctagagtt cagaagaact cgtcaagaag gcgatagaag gcgatgcgct
     4561 gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa
     4621 gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca
     4681 gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc
     4741 aggcatcgcc gtgggtcacg acgagatcct cgccgtcggg catgcgcgcc ttgagcctgg
     4801 cgaacagttc ggctggcgcg agcccctgat gctcttcgtc cagatcatcc tgatcgacaa
     4861 gaccggcttc catccgagta cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg
     4921 ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt
     4981 tctcggcagg agcaaggtga gatgacagga gatcctgccc cggcacttcg cccaatagca
     5041 gccagtccct tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg
     5101 tggccagcca cgatagccgc gctgcctcgt cctgcagttc attcagggca ccggacaggt
     5161 cggtcttgac aaaaagaacc gggcgcccct gcgctgacag ccggaacacg gcggcatcag
     5221 agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc caagcggccg
     5281 gagaacctgc gtgcaatcca tcttgttcaa tcatggtggc atcgattccc ggtcaacatc
     5341 cagtacacct ccggcacacc ggcttcccca agggcgccac gctctcccac cacaacatcc
     5401 tcaacaacgg ctactgggtg ggccgcacgg tcggctacac ggagcaggac cgggtgtgtc
     5461 tgccggtgcc cttctaccac tgcttcggca tggtcatggg caacttgggc gccacctccc
     5521 acgccgcctg catcgtcatc ccggccccgt ccttcgagcc ggcggcgacg ctggaggcgg
     5581 tgcagcggga gaggtgcacg tccctgtacg gcgtccccac gaagccgaat tccaccacac
     5641 tggactagat gcatgctcga cagtgtgatg gatatctgca gaattcggct ttcacgacac
     5701 ctgaaatgga agaaaaaaac tttgaaccac tgtctgaggc ttgagaatga accaagatcc
     5761 aaactcaaaa agggcaaatt ccaaggagaa ttacatcaag tgccaagctg gcctaacttc
     5821 agtctccacc cactcagtgt ggggaaactc catcgcataa aacccctccc cccaacctaa
     5881 agacgacgta ctccaaaagc tcgagaacta atcgaggtgc ctggacggcg cccggtactc
     5941 cgtggagtca catgaagcga cggctgagga cggaaaggcc cttttccttt gtgtgggtga
     6001 ctcacccgcc cgctctcccg agcgccgcgt cctccatttt gagctgcctg cagcagggcc
     6061 gggaagcggc catctttccg ctcacgcaac tggtgccgac cgggccagcc ttgccgccca
     6121 gggcggggcg atacacggcg gcgcgaggcc aggcaccaga gcaggccggc cagcttgaga
     6181 ctacccccgt ccgattctcg gtggccgcgc tcgcaggccc cgcctcgccg aacatgtgcg
     6241 ctgggacgca cgggccccgt cgccgcccgc ggccccaaaa accgaaatac cagtgtgcag
     6301 atcttggccc gcatttacaa gactatcttg ccagaaaaaa agcgtcgcag caggtcatca
     6361 aaaattttaa atggctagag acttatcgaa agcagcgaga caggcgcgaa ggtgccacca
     6421 gattcgcacg cggcggcccc agcgcccagg ccaggcctca actcaagcac gaggcgaagg
     6481 ggctccttaa gcgcaaggcc tcgaactctc ccacccactt ccaacccgaa gctcgggatc
     6541 aagaatcacg tactgcagcc aggtggaagt aattcaaggc acgcaagggc cataacccgt
     6601 aaagaggcca ggcccgcggg aaccacacac ggcacttacc tgtgttctgg cggcaaaccc
     6661 gttgcgaaaa agaacgttca cggcgactac tgcacttata tacggttctc ccccaccctc
     6721 gggaaaaagg cggagccagt acacgacatc actttcccag tttaccccgc gccaccttct
     6781 ctaggcaccg gttcaattgc cgacccctcc ccccaacttc tcggggactg tgggcgatgt
     6841 gcgctctgcc cactgacggg caccggagcc
//
