LOCUS       sequence-472470-        6145 bp    DNA     circular SYN 13-MAY-2025
DEFINITION  Express YAP-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 6145)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6145)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: NanoBRET Assay Vector, YAP1
FEATURES             Location/Qualifiers
     source          1..6145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1499..2011)
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     primer_bind     complement(2024..2043)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2025..2043)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2085..2104)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          2087..2219
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(2355..2558)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2384..2404)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2559..2938
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(3383..3400)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3517..4062)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(3597..3616)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4219..4267
                     /note="synthetic polyadenylation signal"
     CDS             complement(4331..5125)
                     /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(4442..4461)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5052..5071
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5157..5204)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(5219..5576)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5233..5368
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5385..5576
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5533..5553
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    5821..5942
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5858..5877)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5912..5931
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5979..5997
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
ORIGIN
        1 tggcagccaa aacagactcc atgtcattaa ggatgtcaga actcaaagct tcctgcagac
       61 ttggcatcag ctcctctcct tctatgttca ttccatctcc ttccagtgtt ccaaggtcca
      121 catttgtccc aggaatggct tcaaggtagt ctgggaaacg gttctgctgt gagggcaggg
      181 tgctttggtt gatagtatca cctgtatcca tctcatccac actgttcagg aagtcatctg
      241 gggttcgagg gacactgtag ctgctcatgc ttagtccact gtctgtactc tcatctcgag
      301 agtgataggt gccactgtta aggaaaggat ctgagctatt ggtcgtcatt gttctcaatt
      361 cctgagacat cccgggagaa gacactggat tttgagtccc accatcctgc tccagtgttg
      421 gtaactggct acgcagggct aactcctgac attttggaga atttgctgtg ctgggattga
      481 tattccgcat tgcctgccga agcagttctt gctgtttcag ccgcagcctc tccttctcca
      541 tctgcagttg ctgcagtcgc atctgttgct gctggttgga gttgctgcca cccatgacgc
      601 ctccctgtgg gctctgggga gccaggggtg gtggctgttt cactggagca ctctgactga
      661 ttctctggtt catggcaaaa cgagggtcaa gccttgggtc tagccaagag gtggtcttgt
      721 tcttatggtt tatatagtaa atttctccat cctgagtcat ggcttgttcc catccatcag
      781 gaagaggacc tgaagccgag ttcatcatat tctgctgcac tggtggactg gtgggggctg
      841 tgacgttcat ctgggacagc atggccttcc tggggtcctg ccatgttgtt gtctgatcga
      901 tgtgatttaa gaagtatctc tgaccagaag atgtctttgc catctcccaa cctgctggca
      961 gaggtacatc atcaggtatc tcaaaagaag actgtcgaag atgctgagct gtgggtgtag
     1021 ctgctgggcc agagactact ccagtggggg tcagtgtccc aggagaaaca gctcccaact
     1081 gcagagaagc tggagaggaa tgagctcgaa catgctgtgg agtcagggct cctgcagtgc
     1141 ctgcatcagt actggcctgt cgggagtggg atttgggctc cggcggcttg aagaaggagt
     1201 cgggcagctt ccggagcctc atgggcacgg tctggggcac gttggccgtc ttggggttca
     1261 tgacggcgtt gaagagcgcc tccaggtcgg tctccgagtc cccgcggacg tgcacgatct
     1321 gatgcccggc ggggggtgcc tgcggcgccg cctgggtcgc cgcgggtgcc ggttgcccgg
     1381 gtccggacgg cgggccctgc ccctgcgggg gctgcgaagg cggctgccct tggccctggg
     1441 gggccggttg aggcggcggc tgctgcccgg gatccatggc gatcgcgccg ctcgagcccg
     1501 ccagaatgcg ttcgcacagc cgccagccgg tcactccgtt gatggttact cggaacagca
     1561 gggagccgtc ggggttgatc aggcgctcgt cgataatttt gttgccgttc cacagggtcc
     1621 ctgttacagt gatctttttg ccgtcgaaca cggcgatgcc ttcatacggc cgtccgaaat
     1681 agtcgatcat gttcggcgta accccgtcga ttaccagtgt gccatagtgc aggatcacct
     1741 taaagtgatg atcatccaca gggtacacca ccttaaaaat tttttcgatc tggcccattt
     1801 ggtcgccgct cagaccttca tacgggatga tgacatggat gtcgatcttc agcccatttt
     1861 caccgctcag gacaatcctt tggatcggag ttacggacac cccgagattc tgaaacaaac
     1921 tggacacacc tccctgttca aggacttggt ccaggttgta gccggctgtc tgtcgccagt
     1981 ccccaacgaa atcttcgagt gtgaagacca tggtgagcgc tagccctata gtgagtcgta
     2041 ttaatactct agccttaaga gctgtaattg aactgggagt ggacacctgt ggagagaaag
     2101 gcaaagtgga tgtcagtaag accaataggt gcctatcaga aacgcaagag tcttctctgt
     2161 ctcgacaagc ccagtttcta ttggtctcct taaacctgtc ttgtaacctt gatacttacc
     2221 tgcccagtgc ctcacgacca acttctgcag cttaagttcg agactgttgt gtcagaagca
     2281 ctgactgcgt tagcaattta actgtgataa actaccgcaa taaagcttct agtgatctga
     2341 cggttcacta aaccagctct gcttatatag acctcccacc gtacacgcct accgcccatt
     2401 tgcgtcaacg gggcggggtt attacgacat tttggaaagt cccgttgatt ttggtgccaa
     2461 aacaaactcc cattgacgtc aatggggtgg agacttggaa atccccgtga gtcaaaccgc
     2521 tatccacgcc cattggtgta ctgccaaaac cgcatcacca tggtaatagc gatgactaat
     2581 acgtagatgt actgccaagt aggaaagtcc cgtaaggtca tgtactgggc ataatgccag
     2641 gcgggccatt taccgtcatt gacgtcaata gggggcggac ttggcatatg atacacttga
     2701 tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc
     2761 cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg
     2821 ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg
     2881 ctatgaacta atgaccccgt aattgattac tattaataac tagtcaataa tcaatgccaa
     2941 catggcggtc atattggaca tgagccaata taaatgtaca tattatgata tagatacaac
     3001 gtatgcaatg gccaatagcc aatattgatt tatgctatat aaccaatgaa taatatggct
     3061 aatggccaat attgaagatc tcgatcctct acgccggacg catcgtggcc ggagtactgc
     3121 ggtttatctc gatggctacg agggcagaca gtaagtggat ttaccataat cccttaattg
     3181 tacgcaccgc taaaacgcgt aatctcatga cggggtggtg cgtaacggca aaagcaccgc
     3241 cggacatcag cgctagcgga gtgtatactg gcttactatg ttggcactga tgagggtgtc
     3301 agtgaagtgc ttcatgtggc aggagaaaaa aggctgcacc ggtgcgtcag cagaatatgt
     3361 gatacaggat atattccgct tcctcgctca ctgactcgct acgctcggtc gttcgactgc
     3421 ggcgagcgga aatggcttac gaacggggcg gagatttcct ggaagatgcc aggaagatac
     3481 ttaacaggga agtgagaggg ccgcggcaaa gccgtttttc cataggctcc gcccccctga
     3541 caagcatcac gaaatctgac gctcaaatca gtggtggcga aacccgacag gactataaag
     3601 ataccaggcg tttccccctg gcggctccct cgtgcgctct cctgttcctg cctttcggtt
     3661 taccggtgtc attccgctgt tatggccgcg tttgtctcat tccacgcctg acactcagtt
     3721 ccgggtaggc agttcgctcc aagctggact gtatgcacga accccccgtt cagtccgacc
     3781 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggaaagacat gcaaaagcac
     3841 cactggcagc agccactggt aattgattta gaggagttag tcttgaagtc atgcgccggt
     3901 taaggctaaa ctgaaaggac aagttttggt gactgcgctc ctccaagcca gttacctcgg
     3961 ttcaaagagt tggtagctca gagaaccttc gaaaaaccgc cctgcaaggc ggttttttcg
     4021 ttttcagagc aagagattac gcgcagacca aaacgatctc aagaagatca tcttattaat
     4081 cagataaaat atttctagat ttcagtgcaa tttatctctt caaatgtagc acctgaagtc
     4141 agccccatac gatataagtt gtaattctca tgtttgacag cttatcatcg catgagcgga
     4201 tccttatcgc tatcgattca cacaaaaaac caacacacag atgtaatgaa aataaagata
     4261 ttttattgcg gccatcgtga tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc
     4321 agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg
     4381 gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag
     4441 caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac
     4501 agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc
     4561 catgggtcac gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt
     4621 cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt
     4681 ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag
     4741 ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag
     4801 gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc
     4861 ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc
     4921 acgatagccg cgctgcctcg tcctgcagtt cattcagggc accggacagg tcggtcttga
     4981 caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga
     5041 ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg
     5101 cgtgcaatcc atcttgttca atcatgatct gcctcctggg tttagttcct caccttgtcg
     5161 tattatacta tgccgatata ctatgccgat gattaattgt caacagttaa ttaagctttt
     5221 tgcaaaagcc taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg
     5281 aggcggcctc ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg
     5341 cggaactggg cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg
     5401 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
     5461 acctggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg
     5521 gggactttcc acaccctaac tgacacacat tccacagctg gttctttccg cctcagaagg
     5581 tacctaacca agttcctctt tcagaggtta tttcaggcca tggtgctgcg cagatccgcg
     5641 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggaaattgta
     5701 agcgttaata ttttgttaaa attcgatttt accacatttg tagaggtttt acttgcttaa
     5761 aaaaacctcc cacatctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     5821 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     5881 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     5941 tatcatgtct gctcgaagcg gccgccccaa ggggttatgc tagttattgc tcagcggtgg
     6001 cagcagccaa ctcagcttcc tttcgggctt tgttagcagc cggatcagct tgcatgcctg
     6061 caggtcgact ctagaggatc cccgggtacc gagcccgaat tcgtttaaac taaccatgta
     6121 agaaagcttt ctttatctag cttgg
//
