LOCUS       sequence-481958-        6844 bp    DNA     circular SYN 04-AUG-2025
DEFINITION  Express TEAD4-NanoLuc 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 6844)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6844)
  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            "Unpublished"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: TEAD4-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6844
                     /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(2149..2167)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2150..2167)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        2287..2590
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2745..2765
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        2798..2845
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2877..3536
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2943..2962)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      complement(3708..4296)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3788..3807)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4422..4470
                     /note="synthetic polyadenylation signal"
     CDS             complement(4494..5288)
                     /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(4605..4624)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5215..5234
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5666..6844)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          5675..6613
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(5698..5718)
                     /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 agagaaactt ctttcaccag cctgtagatg tggtgctgag
      841 ccccgtgctc actggctgac acctcaaaga cataggcaat gcacagcaag gtctcctgtg
      901 tgtctctgtt ggtgaccacc tgcaggatgg tgaagttctc cagcacgctg ttcatcatgt
      961 acttctcagg gaggtgcttg agcttgtgga tgaagttgat catgtactca cagagcgggg
     1021 accggtggat gcggtaagag tagtgtccat tctcatagcg agcatactct gtctcaactt
     1081 tctccaccac ctgcttgccg aaagagcaga ccttcgtgga gcaggtgatg atcatgttct
     1141 cggggctctc atactggctg gagaccccat agaaggagct gccttcatcc tcgatgttgg
     1201 tgttgaggtc tgcccagaac ttcacaagaa aaaaggcatt ggagggtccc cgttcgaaga
     1261 gatccttgag tccacccttt ttctccggga atttgtcata gatttggcgg atgtccacgg
     1321 cttcgaggta ggggtcgctg tagcttgggc tggactggcc aatgtgcacg aacaggtgct
     1381 tgttgtacgt gtccgggtcc tgctgctgct ccaggaaggc agagaactcc aacatccaga
     1441 gcttggagct ggccacgctg cggccctgcc atgggggtgc cgggggcgca gagggcgatg
     1501 gggcgggccc tgcaggagac tcaaaccctg gcagaggcag cggaggctgg acagcatagg
     1561 tttgctgaga gaaaggcttc acatcatggg acgttccggc ttggcctggc aaagctcctt
     1621 gccaaaaccc tgagactgct gggcggccgg ggccccgggc gagggccatg ctactgtgga
     1681 aggccgtggc ggagatgatc tgtgcagacg acatggcagc catgctctgc agggccttgt
     1741 ccttagctgc ctggtccttt agcttggcct ggatctcgcg agctttgcga cgagccagca
     1801 cctggatgtg gctggagacc tgcttcctgg tgcgggtctt ccctgtccgg agcttgatgt
     1861 agcgggcaat cagctcgttc cgaccataca tcttgccctc gtccgacagg atgattttgc
     1921 gcctgccaca gggcgggtag atggcgaggg cctcctggaa actctgctca atatccgggc
     1981 tccacacgcc ctctgcgtca ttgtcgatgg gcttgtccag tgcctgactg cccccagagg
     2041 cggtgctccc ctcaggggag gtgggagagc tccactcgtt ggaggtaatg gtgccggccg
     2101 tgccctccat ggcgatcgct agctgcaaaa agaacaagta gcttgtattc tatagtgtca
     2161 cctaaatggg cgatctgacg gttcactaaa ccagctctgc ttatataggc ctccctggcg
     2221 cgccactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     2281 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     2341 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     2401 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     2461 tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     2521 ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     2581 tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc
     2641 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     2701 tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag
     2761 gcgtgtacgg tgggaggtca aatcgaattc taatactgtt gacaattaat catcggcata
     2821 gtatatcggc atagtataat acgacaaggt gaggaactaa acccaggagg cagatcatgg
     2881 agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt
     2941 ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta
     3001 cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca
     3061 ttcttgcccg cctgatgaat gctcatccgg aattccgtat ggcaatgaaa gacggtgagc
     3121 tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt
     3181 tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc
     3241 aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata
     3301 tgtttttcgt ctcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca
     3361 atatggacaa cttcttcgcc cccgttttca ctatgggcaa atattatacg caaggcgaca
     3421 aggtgctgat gccgctggcg attcaggttc atcatgccgt ctgtgatggc ttccatgtcg
     3481 gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaagcgg
     3541 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac cggtatcagc tcactcaaag
     3601 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     3661 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3721 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3781 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3841 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3901 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3961 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4021 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4081 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4141 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4201 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4261 aagcagcaga ttacgcgcag aaaaaaagga tttcaagaag atcctttgat cttttctacg
     4321 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4381 aaaaggatct tcacctagat ccttttatag tccggaatcg tcacacaaaa aaccaacaca
     4441 cagatgtaat gaaaataaag atattttatt gcggccatcg tgatggctag agttcagaag
     4501 aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa
     4561 agcacgagga agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc
     4621 aacgctatgt cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa
     4681 aagcggccat tttccaccat gatattcggc aagcaggcat cgccgtgggt cacgacgaga
     4741 tcctcgccgt cgggcatgcg cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc
     4801 tgatgctctt cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct
     4861 cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc
     4921 agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac
     4981 aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca
     5041 acgtcgagca cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc
     5101 tcgtcctgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc
     5161 ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag
     5221 tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt
     5281 tcaatcatgg tggcatcgat tcccggtcaa catccagtac acctccggca caccggcttc
     5341 cccaagggcg ccacgctctc ccaccacaac atcctcaaca acggctactg ggtgggccgc
     5401 acggtcggct acacggagca ggaccgggtg tgtctgccgg tgcccttcta ccactgcttc
     5461 ggcatggtca tgggcaactt gggcgccacc tcccacgccg cctgcatcgt catcccggcc
     5521 ccgtccttcg agccggcggc gacgctggag gcggtgcagc gggagaggtg cacgtccctg
     5581 tacggcgtcc ccacgaagcc gaattccacc acactggact agatgcatgc tcgacagtgt
     5641 gatggatatc tgcagaattc ggctttcacg acacctgaaa tggaagaaaa aaactttgaa
     5701 ccactgtctg aggcttgaga atgaaccaag atccaaactc aaaaagggca aattccaagg
     5761 agaattacat caagtgccaa gctggcctaa cttcagtctc cacccactca gtgtggggaa
     5821 actccatcgc ataaaacccc tccccccaac ctaaagacga cgtactccaa aagctcgaga
     5881 actaatcgag gtgcctggac ggcgcccggt actccgtgga gtcacatgaa gcgacggctg
     5941 aggacggaaa ggcccttttc ctttgtgtgg gtgactcacc cgcccgctct cccgagcgcc
     6001 gcgtcctcca ttttgagctg cctgcagcag ggccgggaag cggccatctt tccgctcacg
     6061 caactggtgc cgaccgggcc agccttgccg cccagggcgg ggcgatacac ggcggcgcga
     6121 ggccaggcac cagagcaggc cggccagctt gagactaccc ccgtccgatt ctcggtggcc
     6181 gcgctcgcag gccccgcctc gccgaacatg tgcgctggga cgcacgggcc ccgtcgccgc
     6241 ccgcggcccc aaaaaccgaa ataccagtgt gcagatcttg gcccgcattt acaagactat
     6301 cttgccagaa aaaaagcgtc gcagcaggtc atcaaaaatt ttaaatggct agagacttat
     6361 cgaaagcagc gagacaggcg cgaaggtgcc accagattcg cacgcggcgg ccccagcgcc
     6421 caggccaggc ctcaactcaa gcacgaggcg aaggggctcc ttaagcgcaa ggcctcgaac
     6481 tctcccaccc acttccaacc cgaagctcgg gatcaagaat cacgtactgc agccaggtgg
     6541 aagtaattca aggcacgcaa gggccataac ccgtaaagag gccaggcccg cgggaaccac
     6601 acacggcact tacctgtgtt ctggcggcaa acccgttgcg aaaaagaacg ttcacggcga
     6661 ctactgcact tatatacggt tctcccccac cctcgggaaa aaggcggagc cagtacacga
     6721 catcactttc ccagtttacc ccgcgccacc ttctctaggc accggttcaa ttgccgaccc
     6781 ctccccccaa cttctcgggg actgtgggcg atgtgcgctc tgcccactga cgggcaccgg
     6841 agcc
//
