LOCUS       sequence-473409-        6145 bp    DNA     circular SYN 06-JUN-2025
DEFINITION  Express NanoLuc(R)-YAP 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 Jun 6, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: NanoLuc-YAP Vector
FEATURES             Location/Qualifiers
     source          1..6145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1065..1577
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     primer_bind     complement(3224..3242)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3279..3400
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3290..3309)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3344..3363
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3645..4002
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3645..3836
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3668..3688)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3853..3988
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4017..4064
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4096..4890
                     /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(4150..4169)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4760..4779
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4954..5002
                     /note="synthetic polyadenylation signal"
     rep_origin      5159..5704
                     /direction=RIGHT
                     /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     5605..5624
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5821..5838
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcgct caccatggtc ttcacactcg
     1081 aagatttcgt tggggactgg cgacagacag ccggctacaa cctggaccaa gtccttgaac
     1141 agggaggtgt gtccagtttg tttcagaatc tcggggtgtc cgtaactccg atccaaagga
     1201 ttgtcctgag cggtgaaaat gggctgaaga tcgacatcca tgtcatcatc ccgtatgaag
     1261 gtctgagcgg cgaccaaatg ggccagatcg aaaaaatttt taaggtggtg taccctgtgg
     1321 atgatcatca ctttaaggtg atcctgcact atggcacact ggtaatcgac ggggttacgc
     1381 cgaacatgat cgactatttc ggacggccgt atgaaggcat cgccgtgttc gacggcaaaa
     1441 agatcactgt aacagggacc ctgtggaacg gcaacaaaat tatcgacgag cgcctgatca
     1501 accccgacgg ctccctgctg ttccgagtaa ccatcaacgg agtgaccggc tggcggctgt
     1561 gcgaacgcat tctggcgggc tcgagcggcg cgatcgccat ggatcccggg cagcagccgc
     1621 cgcctcaacc ggccccccag ggccaagggc agccgccttc gcagcccccg caggggcagg
     1681 gcccgccgtc cggacccggg caaccggcac ccgcggcgac ccaggcggcg ccgcaggcac
     1741 cccccgccgg gcatcagatc gtgcacgtcc gcggggactc ggagaccgac ctggaggcgc
     1801 tcttcaacgc cgtcatgaac cccaagacgg ccaacgtgcc ccagaccgtg cccatgaggc
     1861 tccggaagct gcccgactcc ttcttcaagc cgccggagcc caaatcccac tcccgacagg
     1921 ccagtactga tgcaggcact gcaggagccc tgactccaca gcatgttcga gctcattcct
     1981 ctccagcttc tctgcagttg ggagctgttt ctcctgggac actgaccccc actggagtag
     2041 tctctggccc agcagctaca cccacagctc agcatcttcg acagtcttct tttgagatac
     2101 ctgatgatgt acctctgcca gcaggttggg agatggcaaa gacatcttct ggtcagagat
     2161 acttcttaaa tcacatcgat cagacaacaa catggcagga ccccaggaag gccatgctgt
     2221 cccagatgaa cgtcacagcc cccaccagtc caccagtgca gcagaatatg atgaactcgg
     2281 cttcaggtcc tcttcctgat ggatgggaac aagccatgac tcaggatgga gaaatttact
     2341 atataaacca taagaacaag accacctctt ggctagaccc aaggcttgac cctcgttttg
     2401 ccatgaacca gagaatcagt cagagtgctc cagtgaaaca gccaccaccc ctggctcccc
     2461 agagcccaca gggaggcgtc atgggtggca gcaactccaa ccagcagcaa cagatgcgac
     2521 tgcagcaact gcagatggag aaggagaggc tgcggctgaa acagcaagaa ctgcttcggc
     2581 aggcaatgcg gaatatcaat cccagcacag caaattctcc aaaatgtcag gagttagccc
     2641 tgcgtagcca gttaccaaca ctggagcagg atggtgggac tcaaaatcca gtgtcttctc
     2701 ccgggatgtc tcaggaattg agaacaatga cgaccaatag ctcagatcct ttccttaaca
     2761 gtggcaccta tcactctcga gatgagagta cagacagtgg actaagcatg agcagctaca
     2821 gtgtccctcg aaccccagat gacttcctga acagtgtgga tgagatggat acaggtgata
     2881 ctatcaacca aagcaccctg ccctcacagc agaaccgttt cccagactac cttgaagcca
     2941 ttcctgggac aaatgtggac cttggaacac tggaaggaga tggaatgaac atagaaggag
     3001 aggagctgat gccaagtctg caggaagctt tgagttctga catccttaat gacatggagt
     3061 ctgttttggc tgccaccaag ctagataaag aaagctttct tacatggtta gtttaaacga
     3121 attcgggctc ggtacccggg gatcctctag agtcgacctg caggcatgca agctgatccg
     3181 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta
     3241 gcataacccc ttggggcggc cgcttcgagc agacatgata agatacattg atgagtttgg
     3301 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     3361 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     3421 ttttatgttt caggttcagg gggagatgtg ggaggttttt ttaagcaagt aaaacctcta
     3481 caaatgtggt aaaatcgaat tttaacaaaa tattaacgct tacaatttcc tgatgcggta
     3541 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgcggatctg cgcagcacca
     3601 tggcctgaaa taacctctga aagaggaact tggttaggta ccttctgagg cggaaagaac
     3661 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3721 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3781 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3841 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3901 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     3961 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagcttaa ttaactgttg
     4021 acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa
     4081 cccaggaggc agatcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     4141 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     4201 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     4261 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     4321 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4381 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4441 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4501 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4561 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4621 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4681 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4741 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4801 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4861 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     4921 aagcgacgcc caacctgcca tcacgatggc cgcaataaaa tatctttatt ttcattacat
     4981 ctgtgtgttg gttttttgtg tgaatcgata gcgataagga tccgctcatg cgatgataag
     5041 ctgtcaaaca tgagaattac aacttatatc gtatggggct gacttcaggt gctacatttg
     5101 aagagataaa ttgcactgaa atctagaaat attttatctg attaataaga tgatcttctt
     5161 gagatcgttt tggtctgcgc gtaatctctt gctctgaaaa cgaaaaaacc gccttgcagg
     5221 gcggtttttc gaaggttctc tgagctacca actctttgaa ccgaggtaac tggcttggag
     5281 gagcgcagtc accaaaactt gtcctttcag tttagcctta accggcgcat gacttcaaga
     5341 ctaactcctc taaatcaatt accagtggct gctgccagtg gtgcttttgc atgtctttcc
     5401 gggttggact caagacgata gttaccggat aaggcgcagc ggtcggactg aacggggggt
     5461 tcgtgcatac agtccagctt ggagcgaact gcctacccgg aactgagtgt caggcgtgga
     5521 atgagacaaa cgcggccata acagcggaat gacaccggta aaccgaaagg caggaacagg
     5581 agagcgcacg agggagccgc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     5641 tcgccaccac tgatttgagc gtcagatttc gtgatgcttg tcaggggggc ggagcctatg
     5701 gaaaaacggc tttgccgcgg ccctctcact tccctgttaa gtatcttcct ggcatcttcc
     5761 aggaaatctc cgccccgttc gtaagccatt tccgctcgcc gcagtcgaac gaccgagcgt
     5821 agcgagtcag tgagcgagga agcggaatat atcctgtatc acatattctg ctgacgcacc
     5881 ggtgcagcct tttttctcct gccacatgaa gcacttcact gacaccctca tcagtgccaa
     5941 catagtaagc cagtatacac tccgctagcg ctgatgtccg gcggtgcttt tgccgttacg
     6001 caccaccccg tcatgagatt acgcgtttta gcggtgcgta caattaaggg attatggtaa
     6061 atccacttac tgtctgccct cgtagccatc gagataaacc gcagtactcc ggccacgatg
     6121 cgtccggcgt agaggatcga gatct
//
