LOCUS       sequence-474691-        6847 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express TEAD3-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 6847)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6847)
  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            "Unpublished"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: TEAD3-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6847
                     /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(275..787)
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     promoter        complement(2152..2170)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2153..2170)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        2290..2593
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2748..2768
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        2801..2848
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2880..3539
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2946..2965)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      complement(3711..4299)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3791..3810)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4425..4473
                     /note="synthetic polyadenylation signal"
     CDS             complement(4497..5291)
                     /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(4608..4627)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5218..5237
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5669..6847)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          5678..6616
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(5701..5721)
                     /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 ctagagtcgc ggccttacgc cagaatgcgt tcgcacagcc
      301 gccagccggt cactccgttg atggttactc ggaacagcag ggagccgtcg gggttgatca
      361 ggcgctcgtc gataattttg ttgccgttcc acagggtccc tgttacagtg atctttttgc
      421 cgtcgaacac ggcgatgcct tcatacggcc gtccgaaata gtcgatcatg ttcggcgtaa
      481 ccccgtcgat taccagtgtg ccatagtgca ggatcacctt aaagtgatga tcatccacag
      541 ggtacaccac cttaaaaatt ttttcgatct ggcccatttg gtcgccgctc agaccttcat
      601 acgggatgat gacatggatg tcgatcttca gcccattttc accgctcagg acaatccttt
      661 ggatcggagt tacggacacc ccgagattct gaaacaaact ggacacacct ccctgttcaa
      721 ggacttggtc caggttgtag ccggctgtct gtcgccagtc cccaacgaaa tcttcgagtg
      781 tgaagacgcc gctcgagccg agagaaacgt ctttgacgag cttgtagaca tggtgctggg
      841 ccccgtgctc actggtggag acttcgaaga caaaagcaat gacaagcagg gtctcctggg
      901 agtcccggct cgtgaccacc tgcaggatgg tgaagttctc cagcacgctg ttcatcatgt
      961 acttctcggg caggtgcttc agcttgtgga tgaagttgat catgtactcg cacatgggcg
     1021 agcggtggat acggtacaca aagcgcccgt tctccagcct ggcatactca gtctccacct
     1081 tctctaccac ctgtttgcca aaggagcaca ccttggtgga gacgctgatg gtcatgctat
     1141 cagcagagct gtactgagag ctgaccccat agaaggctcc cgggccctcc tggatggtgc
     1201 tgttgaggtc ggcccagaac ttgacaagga agaaggcatt agggggcccc ttctcataga
     1261 gctccttcaa tcctcccttt ttctcgggga atttgtcata gatctggcgc acatctactg
     1321 cctccagggg tgggtctgag aaggcggggt tcgtctggcc gatgtgcaca aacaggtgtt
     1381 tgctgtacgt gtcagggtct cgctgcacct ccatgaaggc tgaatactcc aggagccgca
     1441 gccgggagga ggcaatggta cggtcctgcc acacaggcac agaggcagca gctgagggga
     1501 gcggggccag gggctcataa ctgctgagcg tcggcggcag gggcggctgg atggggtagg
     1561 ctggctgtgc aaagggcttg atgtcctgag agggtccagg ctgctgtccc aggagagggg
     1621 ggctgctcca gaaccgcgag gaagtggaga agacggcctg gggcagaggg gaaggtgggc
     1681 tgaacttgtt ctgcaggaca ctggcagaga cgatctgggc agaggacatg gacgccatgc
     1741 tctgaagggc tttgtccttg gagacctggt ccaggttcat ggccttgatg ccaacctggt
     1801 actcccgcac cttcttccga gctagaacct gtatgtggct ggacacctgt tttctcgtcc
     1861 gagtcttccc cgtcctcagt ttaatatagc gtgcaatcaa ctcatttcgg ccgtacatct
     1921 tgccctcgtc tgacaggatg atcttccgcc ggccgcaggg cgggtagatg gccagggcct
     1981 cctggaagct ctgctcgatg tccgggctcc acacgccctc cgcatcgttg tccagcccct
     2041 tgtccaggcc ctcgggccca tcctcccggg cctccccggg gctgctgctg gcgttccagc
     2101 tgttggacgc catggcgatc gctagctgca aaaagaacaa gtagcttgta ttctatagtg
     2161 tcacctaaat gggcgatctg acggttcact aaaccagctc tgcttatata ggcctccctg
     2221 gcgcgccact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     2281 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     2341 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     2401 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     2461 tcatatgcca agtccgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     2521 tgcccagtac atgaccttac gggactttcc tacttggcag tacatctacg tattagtcat
     2581 cgctattacc atggtgatgc ggttttggca gtacaccaat gggcgtggat agcggtttga
     2641 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     2701 aaatcaacgg gactttccaa aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg
     2761 taggcgtgta cggtgggagg tcaaatcgaa ttctaatact gttgacaatt aatcatcggc
     2821 atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaacccagg aggcagatca
     2881 tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac
     2941 attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata
     3001 ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc
     3061 acattcttgc ccgcctgatg aatgctcatc cggaattccg tatggcaatg aaagacggtg
     3121 agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa
     3181 cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt
     3241 cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga
     3301 atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg
     3361 ccaatatgga caacttcttc gcccccgttt tcactatggg caaatattat acgcaaggcg
     3421 acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtctgtgat ggcttccatg
     3481 tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaag
     3541 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aaccggtatc agctcactca
     3601 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3661 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3721 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3781 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3841 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3901 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3961 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4021 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4081 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4141 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4201 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4261 tgcaagcagc agattacgcg cagaaaaaaa ggatttcaag aagatccttt gatcttttct
     4321 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4381 tcaaaaagga tcttcaccta gatcctttta tagtccggaa tcgtcacaca aaaaaccaac
     4441 acacagatgt aatgaaaata aagatatttt attgcggcca tcgtgatggc tagagttcag
     4501 aagaactcgt caagaaggcg atagaaggcg atgcgctgcg aatcgggagc ggcgataccg
     4561 taaagcacga ggaagcggtc agcccattcg ccgccaagct cttcagcaat atcacgggta
     4621 gccaacgcta tgtcctgata gcggtccgcc acacccagcc ggccacagtc gatgaatcca
     4681 gaaaagcggc cattttccac catgatattc ggcaagcagg catcgccgtg ggtcacgacg
     4741 agatcctcgc cgtcgggcat gcgcgccttg agcctggcga acagttcggc tggcgcgagc
     4801 ccctgatgct cttcgtccag atcatcctga tcgacaagac cggcttccat ccgagtacgt
     4861 gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta
     4921 tgcagccgcc gcattgcatc agccatgatg gatactttct cggcaggagc aaggtgagat
     4981 gacaggagat cctgccccgg cacttcgccc aatagcagcc agtcccttcc cgcttcagtg
     5041 acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct
     5101 gcctcgtcct gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg
     5161 cgcccctgcg ctgacagccg gaacacggcg gcatcagagc agccgattgt ctgttgtgcc
     5221 cagtcatagc cgaatagcct ctccacccaa gcggccggag aacctgcgtg caatccatct
     5281 tgttcaatca tggtggcatc gattcccggt caacatccag tacacctccg gcacaccggc
     5341 ttccccaagg gcgccacgct ctcccaccac aacatcctca acaacggcta ctgggtgggc
     5401 cgcacggtcg gctacacgga gcaggaccgg gtgtgtctgc cggtgccctt ctaccactgc
     5461 ttcggcatgg tcatgggcaa cttgggcgcc acctcccacg ccgcctgcat cgtcatcccg
     5521 gccccgtcct tcgagccggc ggcgacgctg gaggcggtgc agcgggagag gtgcacgtcc
     5581 ctgtacggcg tccccacgaa gccgaattcc accacactgg actagatgca tgctcgacag
     5641 tgtgatggat atctgcagaa ttcggctttc acgacacctg aaatggaaga aaaaaacttt
     5701 gaaccactgt ctgaggcttg agaatgaacc aagatccaaa ctcaaaaagg gcaaattcca
     5761 aggagaatta catcaagtgc caagctggcc taacttcagt ctccacccac tcagtgtggg
     5821 gaaactccat cgcataaaac ccctcccccc aacctaaaga cgacgtactc caaaagctcg
     5881 agaactaatc gaggtgcctg gacggcgccc ggtactccgt ggagtcacat gaagcgacgg
     5941 ctgaggacgg aaaggccctt ttcctttgtg tgggtgactc acccgcccgc tctcccgagc
     6001 gccgcgtcct ccattttgag ctgcctgcag cagggccggg aagcggccat ctttccgctc
     6061 acgcaactgg tgccgaccgg gccagccttg ccgcccaggg cggggcgata cacggcggcg
     6121 cgaggccagg caccagagca ggccggccag cttgagacta cccccgtccg attctcggtg
     6181 gccgcgctcg caggccccgc ctcgccgaac atgtgcgctg ggacgcacgg gccccgtcgc
     6241 cgcccgcggc cccaaaaacc gaaataccag tgtgcagatc ttggcccgca tttacaagac
     6301 tatcttgcca gaaaaaaagc gtcgcagcag gtcatcaaaa attttaaatg gctagagact
     6361 tatcgaaagc agcgagacag gcgcgaaggt gccaccagat tcgcacgcgg cggccccagc
     6421 gcccaggcca ggcctcaact caagcacgag gcgaaggggc tccttaagcg caaggcctcg
     6481 aactctccca cccacttcca acccgaagct cgggatcaag aatcacgtac tgcagccagg
     6541 tggaagtaat tcaaggcacg caagggccat aacccgtaaa gaggccaggc ccgcgggaac
     6601 cacacacggc acttacctgt gttctggcgg caaacccgtt gcgaaaaaga acgttcacgg
     6661 cgactactgc acttatatac ggttctcccc caccctcggg aaaaaggcgg agccagtaca
     6721 cgacatcact ttcccagttt accccgcgcc accttctcta ggcaccggtt caattgccga
     6781 cccctccccc caacttctcg gggactgtgg gcgatgtgcg ctctgcccac tgacgggcac
     6841 cggagcc
//
