LOCUS       sequence-475172-        7001 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express PPP2R5A-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 7001)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7001)
  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            "Unpublished"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: NanoBRET Assay Vector, PPP2R5A-NanoLuc
FEATURES             Location/Qualifiers
     source          1..7001
                     /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(2305..2323)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2306..2323)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     enhancer        2443..2746
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2901..2921
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        2954..3001
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3033..3692
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3099..3118)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      complement(3864..4452)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3944..3963)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4578..4626
                     /note="synthetic polyadenylation signal"
     CDS             complement(4650..5444)
                     /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(4761..4780)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5371..5390
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5823..7001)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          5832..6770
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(5855..5875)
                     /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 cggcacttgt attgctgaga atactgtgca
      841 tgttgtaagc actattctgt ttttctagag ctttctttag ctttagctcc tctaattttt
      901 tccataattc ttcacgttcc aattccttct ttttctctct ctgtctttca gctttgtatg
      961 agctagtaag gtcatcgaaa agcttgccat tcatttccat tagggttttc agcacattgt
     1021 ataccagtgc tacaatggtc ggattccagt gttctttgga aattttgtac aaactggcaa
     1081 acataattgg cagaatttta tcaatgttct cctcaatcaa actaagaata tattcgttat
     1141 tccagaagta caatgccctt tctgcaacct gaaaatgaga actggataca cacttggata
     1201 tctgcttgaa aagtggctct tcaatttttt tgaactgtgt tggttcaatg acatctaaga
     1261 tttcttcaat ttctcctaaa aacatcacct ctttctgact gcaggttttt ggccaaaatt
     1321 tcagcagtcc tctgatcact ggctctgtta gtgttgtatc tttctccagg aactgtacaa
     1381 cacaatatgc tagctgagca tgaaacaaag ctaatccttt tgcagtatgc ataggaataa
     1441 gaaccttcat tagaaattgt ttatgttctg ctttcagtgg caatgcaaag ccattgataa
     1501 tacttcctaa tatttcaaga agttcagcaa caccattgaa atgttctgtt tcatatataa
     1561 acctgaggaa aatgttgtta atttgttttc tgatgaatgc tcttaatcca agaaatttcc
     1621 cataaattcg gtgcagaaca gtcttcagga agtcacgttc tctgggatct tcactatcaa
     1681 aaagctccag gagctgttgt acgaatttct gatcaatgta tcgttttgca atgctaggct
     1741 ggaaatcagg gctctccaaa aatctcaaga agaattcata taccaactgt atgtgaggcc
     1801 aagaggcctc aagcgtgggt tcatcctctt ctggatcaaa atctggatta tcacttggag
     1861 gaagtgtacg gaagatgtta gcactgatca tttttactat atcagaatac gctgattcaa
     1921 caattacacc acgattagtt gaaacatact caaccagttc attcagtgtt gctcttttaa
     1981 tttctttgct cttcaagtct gaaacagagt ccatgaaatc aaacagtata caacactgct
     2041 gcaacttctg acagaaaagc tcttgttgtt catttgaagt ggcatctttg agctggggca
     2101 gcgggtgcag ctctgcctgg ctgccctggc tgcgaaactg cgacgagccc tgggagcgct
     2161 tctgcctctg cgccttgcgg accgatttcc gggtgaagcc gtccactttc tccgaggccg
     2221 agatggcggc gctggcagcc cccgccggcg gcgacgacga cgacatggcg atcgctagct
     2281 gcaaaaagaa caagtagctt gtattctata gtgtcaccta aatgggcgat ctgacggttc
     2341 actaaaccag ctctgcttat ataggcctcc ctggcgcgcc actagttatt aatagtaatc
     2401 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     2461 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     2521 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     2581 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtccgc cccctattga
     2641 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tacgggactt
     2701 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
     2761 gcagtacacc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     2821 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     2881 taataacccc gccccgttga cgcaaatggg cggtaggcgt gtacggtggg aggtcaaatc
     2941 gaattctaat actgttgaca attaatcatc ggcatagtat atcggcatag tataatacga
     3001 caaggtgagg aactaaaccc aggaggcaga tcatggagaa aaaaatcact ggatatacca
     3061 ccgttgatat atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc
     3121 aatgtaccta taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga
     3181 aaaataagca caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc
     3241 atccggaatt ccgtatggca atgaaagacg gtgagctggt gatatgggat agtgttcacc
     3301 cttgttacac cgttttccat gagcaaactg aaacgttttc atcgctctgg agtgaatacc
     3361 acgacgattt ccggcagttt ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa
     3421 acctggccta tttccctaaa gggtttattg agaatatgtt tttcgtctca gccaatccct
     3481 gggtgagttt caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg
     3541 ttttcactat gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc
     3601 aggttcatca tgccgtctgt gatggcttcc atgtcggcag aatgcttaat gaattacaac
     3661 agtactgcga tgagtggcag ggcggggcgt aagcgggact ctggggttcg aaatgaccga
     3721 ccaagcgacg cccaaccggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3781 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3841 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3901 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3961 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4021 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4081 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4141 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4201 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4261 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     4321 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4381 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4441 aaaggatttc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4501 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4561 ttatagtccg gaatcgtcac acaaaaaacc aacacacaga tgtaatgaaa ataaagatat
     4621 tttattgcgg ccatcgtgat ggctagagtt cagaagaact cgtcaagaag gcgatagaag
     4681 gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat
     4741 tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc
     4801 gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata
     4861 ttcggcaagc aggcatcgcc gtgggtcacg acgagatcct cgccgtcggg catgcgcgcc
     4921 ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc cagatcatcc
     4981 tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg tttcgcttgg
     5041 tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg
     5101 atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc cggcacttcg
     5161 cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga
     5221 acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cctgcagttc attcagggca
     5281 ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag ccggaacacg
     5341 gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc
     5401 caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcatggtggc atcgattccc
     5461 ggtcaacatc cagtacacct ccggcaccac cggcttcccc aagggcgcca cgctctccca
     5521 ccacaacatc ctcaacaacg gctactgggt gggccgcacg gtcggctaca cggagcagga
     5581 ccgggtgtgt ctgccggtgc ccttctacca ctgcttcggc atggtcatgg gcaacttggg
     5641 cgccacctcc cacgccgcct gcatcgtcat cccggccccg tccttcgagc cggcggcgac
     5701 gctggaggcg gtgcagcggg agaggtgcac gtccctgtac ggcgtcccca cgaagccgaa
     5761 ttccaccaca ctggactaga tgcatgctcg acagtgtgat ggatatctgc agaattcggc
     5821 tttcacgaca cctgaaatgg aagaaaaaaa ctttgaacca ctgtctgagg cttgagaatg
     5881 aaccaagatc caaactcaaa aagggcaaat tccaaggaga attacatcaa gtgccaagct
     5941 ggcctaactt cagtctccac ccactcagtg tggggaaact ccatcgcata aaacccctcc
     6001 ccccaaccta aagacgacgt actccaaaag ctcgagaact aatcgaggtg cctggacggc
     6061 gcccggtact ccgtggagtc acatgaagcg acggctgagg acggaaaggc ccttttcctt
     6121 tgtgtgggtg actcacccgc ccgctctccc gagcgccgcg tcctccattt tgagctgcct
     6181 gcagcagggc cgggaagcgg ccatctttcc gctcacgcaa ctggtgccga ccgggccagc
     6241 cttgccgccc agggcggggc gatacacggc ggcgcgaggc caggcaccag agcaggccgg
     6301 ccagcttgag actacccccg tccgattctc ggtggccgcg ctcgcaggcc ccgcctcgcc
     6361 gaacatgtgc gctgggacgc acgggccccg tcgccgcccg cggccccaaa aaccgaaata
     6421 ccagtgtgca gatcttggcc cgcatttaca agactatctt gccagaaaaa aagcgtcgca
     6481 gcaggtcatc aaaaatttta aatggctaga gacttatcga aagcagcgag acaggcgcga
     6541 aggtgccacc agattcgcac gcggcggccc cagcgcccag gccaggcctc aactcaagca
     6601 cgaggcgaag gggctcctta agcgcaaggc ctcgaactct cccacccact tccaacccga
     6661 agctcgggat caagaatcac gtactgcagc caggtggaag taattcaagg cacgcaaggg
     6721 ccataacccg taaagaggcc aggcccgcgg gaaccacaca cggcacttac ctgtgttctg
     6781 gcggcaaacc cgttgcgaaa aagaacgttc acggcgacta ctgcacttat atacggttct
     6841 cccccaccct cgggaaaaag gcggagccag tacacgacat cactttccca gtttaccccg
     6901 cgccaccttc tctaggcacc ggttcaattg ccgacccctc cccccaactt ctcggggact
     6961 gtgggcgatg tgcgctctgc ccactgacgg gcaccggagc c
//
