LOCUS       sequence-482866-        8677 bp    DNA     circular SYN 11-AUG-2025
DEFINITION  Express TEAD2 / YAP Fusion Proteins in Mammalian Cells under a CMV 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8677)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8677)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 11, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: NanoBRET Assay, TEAD2 / YAP Bidirectional Vector
FEATURES             Location/Qualifiers
     source          1..8677
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(747..1541)
                     /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(858..877)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1468..1487
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1573..1620)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     protein_bind    1630..1663
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    1778..1899
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     1869..1888
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(1986..2870)
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     CDS             complement(2886..2906)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     primer_bind     complement(4281..4300)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4282..4300)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4342..4361)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          4344..4476
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(4612..4815)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4641..4661)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4816..5119
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5239..5257
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5239..5256
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             5286..5798
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     polyA_signal    7534..7615
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        7694..8051
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        7694..7885
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(7717..7737)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      7902..8037
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      8076..8085
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             8082..8480
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    8504..8552
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 ttgaaatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccggttg
      721 ggcgtcgctt ggtcggtcat ttcgaatcag aagaactcgt caagaaggcg atagaaggcg
      781 atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg
      841 ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc
      901 acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc
      961 ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gcgcgccttg
     1021 agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga
     1081 tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg
     1141 tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg
     1201 gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc
     1261 aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg
     1321 cccgtcgtgg ccagccacga tagccgcgct gcctcgtcct gcagttcatt cagggcaccg
     1381 gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg
     1441 gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa
     1501 gcggccggag aacctgcgtg caatccatct tgttcaatca tgatctgcct cctgggttta
     1561 gttcctcacc ttgtcgtatt atactatgcc gatatactat gccgatgatt aattgtcaac
     1621 agtggatcca taacttcgta taatgtatgc tatacgaagt tatattagaa ttcttaatta
     1681 agattttacc acatttgtag aggttttact tgcttaaaaa aacctcccac atctccccct
     1741 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     1801 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     1861 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgct cgaagcggcc
     1921 ttgctcagcg gtggcagcag ccaaatcagc ttgcatgcct gcaggtcgac tctagaattc
     1981 tattaaccgg aaatctccag agtagacagc cagcgcgcga tctcgctgcc gatcaggtcc
     2041 gggttgtctt cttgcagcag attcagaccc gggccgatgt ccacagcctt gcagttaggc
     2101 aggcttttgg ccaggcgagc ggcttcggcc ggtgggatca gaacgcctgg ggtgccccag
     2161 aacagcagct tcgggacagg ggactggtgc agccagtcca tgtattcttc gaccagcgcg
     2221 acgatgttcg ctggctcacc ggcgattggc agctcgtttg ggaagcgcca cagtggctcg
     2281 cggtcaacag gattcaggaa cggctcgcgg taatggtcca tctcgacttc agtcagcggg
     2341 cggacgacac ccatcggcag cgtaccctcg ataaaaacgt tctgatcgat gatcagcttg
     2401 cggccgacgt cggtggtgcg gaaggcctgg aaggtctcgc gggcaaattc tggccattcg
     2461 tcccaggtcg ggatagggcg gatgaactcc ataaatgcaa tacctttgac gcgctctgga
     2521 ttgcgcttgg cccagtggaa acccagagcg gagccccagt cgtgaatgac caggacgacc
     2581 tcttccagac ccagggcttc gatgaaggca tccatgaagc ggacgtggtc gtcgaagaaa
     2641 taacccaggt ctggtttgtc ggatttgccc ataccgatca ggtctggagc aatgcagcga
     2701 tgggtcggtg caacatgcgg gatgatgttg cgccacacgt aggaggaggt cgggttaccg
     2761 tgcaggaaca gcacaggggt gccatcgcgc ggaccaacat cgacgtagtg catgcgctcg
     2821 cccaggactt ccacataatg ggggtcgaat ggaaagccag taccgatttc ggatccatcg
     2881 ttatcgctct gaaagtacag atcctcagtg gttggctcga gagaaacgtc cctgaccagg
     2941 cggtaaatgt gatgctgggc cccacgctcg ctggtggaga cctcgaagac ataggcggtg
     3001 cagagcagca gttcctgggt gtctctgttt gtcaccacct ggaggatggt gaagttttcc
     3061 aggacgctgt tcatcatgta tcgctcaggc agctgccgca acttgtgcaa gaaattcacc
     3121 aggtactcgc acatgggcga gcgcagcagg cggtacacaa atctgccgtc ctccagctgg
     3181 gcccgttccg tctccacctt ctccaccacc tgcttgccaa aagagcagac cttggaggaa
     3241 caggtgaggg tcatgtgttc caggctctca tactggctgc tcactccgta gaagccacca
     3301 ctgctgatgc tgccaccggc ccctgcctcc tcaccacttg ggccccagtt caggtccgcc
     3361 cagaacttga ccaggaagaa ggcatggggg gggccacgat catatagctc tcggaggcca
     3421 ccctttttct cagggaattt gtcgtagatc tgccggacgt ccacactctc gagcggcggc
     3481 gctccggggc tggggcagtg ctggctgatg tgcacgaaca ggtgcctctg gtaagaatca
     3541 actgcatctg gcggttccac gaaggctgag aactctacca gctgcaaccg ggcggtgccc
     3601 aggccccgag cctgccaggc tgggggcgat ggggtaggtg ggggcagggg tgagagggct
     3661 tgggggggct cgtaccctgg gaggtcagta gatgggggag tcagtgacaa ggtgaacggt
     3721 gtctgtgaga atggcttcac atctggaaca ttccaggggg gcccagatcc tccagaccaa
     3781 aactggaaaa gctcagaggc ctgaggacca gtgggaccca gtttggcctg cagagaaggc
     3841 gcggagatga gctgggcaga ggacatggtt gccattgtct ggaaagcctt gtccttggaa
     3901 acctggtcct tcaacttgga ctggatttcc cttgatttcc ttcgggccaa aacctggatg
     3961 tgactagaaa cctgttttcg agttcgggtc ttccccgttc tcagcttgat gtagcgggcg
     4021 atcagttcat tccgaccata catcttgcct tcatcagaca aaattatttt ccggcggccg
     4081 cagggtggat agatggccag ggcctcctgg aagctctgct caatgtctgg gctccacacc
     4141 ccctctgcat ccgggccccc gtcaccccca gccccctcac tgccgccggt accctcctca
     4201 ctgccttcct cactgcccgt ccagccgctg ccatcgtcca gggcggcccc agcccggggt
     4261 tcccccatgg cgatcgctag ccctatagtg agtcgtatta atactctagc cttaagagct
     4321 gtaattgaac tgggagtgga cacctgtgga gagaaaggca aagtggatgt cagtaagacc
     4381 aataggtgcc tatcagaaac gcaagagtct tctctgtctc gacaagccca gtttctattg
     4441 gtctccttaa acctgtcttg taaccttgat acttacctgc ccagtgcctc acgaccaact
     4501 tctgcagctt aagttcgaga ctgttgtgtc agaagcactg actgcgttag caatttaact
     4561 gtgataaact accgcaataa agcttctagt gatctgacgg ttcactaaac cagctctgct
     4621 tatatagacc tcccaccgta cacgcctacc gcccatttgc gtcaacgggg cggggttatt
     4681 acgacatttt ggaaagtccc gttgattttg gtgccaaaac aaactcccat tgacgtcaat
     4741 ggggtggaga cttggaaatc cccgtgagtc aaaccgctat ccacgcccat tggtgtactg
     4801 ccaaaaccgc atcaccatgg taatagcgat gactaatacg tagatgtact gccaagtagg
     4861 aaagtcccgt aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac
     4921 gtcaataggg ggcggacttg gcatatgata cacttgatgt actgccaagt gggcagttta
     4981 ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac
     5041 atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt
     5101 taccgtaagt tatgtaacgc ggaactccat atatgggcta tgaactaatg accccgtaat
     5161 tgattactat taataactag tggcgcgcca gggaggccta tatcagcaga gctggtttag
     5221 tgaaccgtca gatcgcccat ttaggtgaca ctatagaata caagctactt gttctttttg
     5281 caaccatggt cttcacactc gaagatttcg ttggggactg gcgacagaca gccggctaca
     5341 acctggacca agtccttgaa cagggaggtg tgtccagttt gtttcagaat ctcggggtgt
     5401 ccgtaactcc gatccaaagg attgtcctga gcggtgaaaa tgggctgaag atcgacatcc
     5461 atgtcatcat cccgtatgaa ggtctgagcg gcgaccaaat gggccagatc gaaaaaattt
     5521 ttaaggtggt gtaccctgtg gatgatcatc actttaaggt gatcctgcac tatggcacac
     5581 tggtaatcga cggggttacg ccgaacatga tcgactattt cggacggccg tatgaaggca
     5641 tcgccgtgtt cgacggcaaa aagatcactg taacagggac cctgtggaac ggcaacaaaa
     5701 ttatcgacga gcgcctgatc aaccccgacg gctccctgct gttccgagta accatcaacg
     5761 gagtgaccgg ctggcggctg tgcgaacgca ttctggcggg ctcgagcggc gcgatcgcca
     5821 tggatcccgg gcagcagccg ccgcctcaac cggcccccca gggccaaggg cagccgcctt
     5881 cgcagccccc gcaggggcag ggcccgccgt ccggacccgg gcaaccggca cccgcggcga
     5941 cccaggcggc gccgcaggca ccccccgccg ggcatcagat cgtgcacgtc cgcggggact
     6001 cggagaccga cctggaggcg ctcttcaacg ccgtcatgaa ccccaagacg gccaacgtgc
     6061 cccagaccgt gcccatgagg ctccggaagc tgcccgactc cttcttcaag ccgccggagc
     6121 ccaaatccca ctcccgacag gccagtactg atgcaggcac tgcaggagcc ctgactccac
     6181 agcatgttcg agctcattcc tctccagctt ctctgcagtt gggagctgtt tctcctggga
     6241 cactgacccc cactggagta gtctctggcc cagcagctac acccacagct cagcatcttc
     6301 gacagtcttc ttttgagata cctgatgatg tacctctgcc agcaggttgg gagatggcaa
     6361 agacatcttc tggtcagaga tacttcttaa atcacatcga tcagacaaca acatggcagg
     6421 accccaggaa ggccatgctg tcccagatga acgtcacagc ccccaccagt ccaccagtgc
     6481 agcagaatat gatgaactcg gcttcaggtc ctcttcctga tggatgggaa caagccatga
     6541 ctcaggatgg agaaatttac tatataaacc ataagaacaa gaccacctct tggctagacc
     6601 caaggcttga ccctcgtttt gccatgaacc agagaatcag tcagagtgct ccagtgaaac
     6661 agccaccacc cctggctccc cagagcccac agggaggcgt catgggtggc agcaactcca
     6721 accagcagca acagatgcga ctgcagcaac tgcagatgga gaaggagagg ctgcggctga
     6781 aacagcaaga actgcttcgg caggcaatgc ggaatatcaa tcccagcaca gcaaattctc
     6841 caaaatgtca ggagttagcc ctgcgtagcc agttaccaac actggagcag gatggtggga
     6901 ctcaaaatcc agtgtcttct cccgggatgt ctcaggaatt gagaacaatg acgaccaata
     6961 gctcagatcc tttccttaac agtggcacct atcactctcg agatgagagt acagacagtg
     7021 gactaagcat gagcagctac agtgtccctc gaaccccaga tgacttcctg aacagtgtgg
     7081 atgagatgga tacaggtgat actatcaacc aaagcaccct gccctcacag cagaaccgtt
     7141 tcccagacta ccttgaagcc attcctggga caaatgtgga ccttggaaca ctggaaggag
     7201 atggaatgaa catagaagga gaggagctga tgccaagtct gcaggaagct ttgagttctg
     7261 acatccttaa tgacatggag tctgttttgg ctgccaccaa gctagataaa gaaagctttc
     7321 ttacatggtt agtttaaacg aattcgggct cggtacccgg ggatcctcta gaaacaccta
     7381 ggctctcgta cgaagtcagc tgataacgtc gacgtattgc ggccgcggat cataatcagc
     7441 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     7501 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     7561 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgccgacg
     7621 tcttaattaa cgcgtcttgc gcagcaccat ggcctgaaat aacctctgaa agaggaactt
     7681 ggttaggtac cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     7741 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     7801 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     7861 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     7921 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     7981 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     8041 gcttttgcaa aaagctcgat tcttctgaca ctagcgccac catggccaag cctttgtctc
     8101 aagaagaatc caccctcatt gaaagagcaa cggctacaat caacagcatc cccatctctg
     8161 aagactacag cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc actggtgtca
     8221 atgtatatca ttttactggg ggaccttgtg cagaactcgt ggtgctgggc actgctgctg
     8281 ctgcggcagc tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac aggggcatct
     8341 tgagcccctg cggacggtgc cgacaggtgc ttctcgatct gcatcctggg atcaaagcca
     8401 tagtgaagga cagtgatgga cagccgacgg cagttgggat tcgtgaattg ctgccctctg
     8461 gttatgtgtg ggagggctaa actctagcca tcacgatggc cgcaataaaa tatctttatt
     8521 ttcattacat ctgtgtgttg gttttttgtg tgacgattcc ggactataaa aggatctagg
     8581 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     8641 gagcgtcaga ccccgtagaa aagatcaaag gatcttc
//
