LOCUS       sequence-486825-        6510 bp    DNA     circular SYN 22-SEP-2025
DEFINITION  Express PPP2R1A-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 6510)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6510)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 22, 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            "Unpublished"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, PPP2R1A-NanoLuc
FEATURES             Location/Qualifiers
     source          1..6510
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             55..567
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(655..673)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    710..831
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(721..740)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     775..794
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1076..1433
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1076..1267
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1099..1119)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1284..1419
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        1448..1495
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1527..2321
                     /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(1581..1600)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2191..2210
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2385..2433
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(2672..3260)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2752..2771)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    3376..3659
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     enhancer        3850..4229
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4230..4433
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4384..4404
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          4569..4701
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     4684..4703
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     4745..4764
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4745..4763
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4775..4784
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 gcccaggagg ctctgactgt tctgtctctc gccgtttctc tcggctcgag cggcgtcttc
       61 acactcgaag atttcgttgg ggactggcga cagacagccg gctacaacct ggaccaagtc
      121 cttgaacagg gaggtgtgtc cagtttgttt cagaatctcg gggtgtccgt aactccgatc
      181 caaaggattg tcctgagcgg tgaaaatggg ctgaagatcg acatccatgt catcatcccg
      241 tatgaaggtc tgagcggcga ccaaatgggc cagatcgaaa aaatttttaa ggtggtgtac
      301 cctgtggatg atcatcactt taaggtgatc ctgcactatg gcacactggt aatcgacggg
      361 gttacgccga acatgatcga ctatttcgga cggccgtatg aaggcatcgc cgtgttcgac
      421 ggcaaaaaga tcactgtaac agggaccctg tggaacggca acaaaattat cgacgagcgc
      481 ctgatcaacc ccgacggctc cctgctgttc cgagtaacca tcaacggagt gaccggctgg
      541 cggctgtgcg aacgcattct ggcgtaaggc cgcgactcta gagtcgacct gcaggcatgc
      601 aagctgatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg
      661 agcaataact agcataaccc cttggggcgg ccgcttcgag cagacatgat aagatacatt
      721 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
      781 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
      841 aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt tttaagcaag
      901 taaaacctct acaaatgtgg taaaatcgaa ttttaacaaa atattaacgc ttacaatttc
      961 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgcggatct
     1021 gcgcagcacc atggcctgaa ataacctctg aaagaggaac ttggttaggt accttctgag
     1081 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     1141 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     1201 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     1261 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     1321 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     1381 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctta
     1441 attaactgtt gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt
     1501 gaggaactaa acccaggagg cagatcatga ttgaacaaga tggattgcac gcaggttctc
     1561 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     1621 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     1681 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     1741 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     1801 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     1861 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     1921 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     1981 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     2041 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     2101 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     2161 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     2221 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     2281 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     2341 aatgaccgac caagcgacgc ccaacctgcc atcacgatgg ccgcaataaa atatctttat
     2401 tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat agcgataagg atcctctttg
     2461 cgcttgcgtt ttcccttgtc cagatagccc agtagctgac attcatccgg ggtcagcacc
     2521 gtttctgcgg actggctttc tacccggtat cagctcactc aaaggcggta atacggttat
     2581 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     2641 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2701 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2761 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2821 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2881 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2941 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3001 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3061 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     3121 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3181 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3241 gcagaaaaaa aggatttcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3301 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3361 agatcctttt atagtccgga aatacaggaa cgcacgctgg atggcccttc gctgggatgg
     3421 tgaaaccatg aaaaatggca gcttcagtgg attaagtggg ggtaatgtgg cctgtaccct
     3481 ctggttgcat aggtattcat acggttaaaa tttatcaggc gcgattgcgg cagtttttcg
     3541 ggtggtttgt tgccattttt acctgtctgc tgccgtgatc gcgctgaacg cgttttagcg
     3601 gtgcgtacaa ttaagggatt atggtaaatc cacttactgt ctgccctcgt agccatcgag
     3661 ataaaccgca gtactccggc cacgatgcgt ccggcgtaga ggatcgagat cttcaatatt
     3721 ggccattagc catattattc attggttata tagcataaat caatattggc tattggccat
     3781 tgcatacgtt gtatctatat cataatatgt acatttatat tggctcatgt ccaatatgac
     3841 cgccatgttg gcattgatta ttgactagtt attaatagta atcaattacg gggtcattag
     3901 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     3961 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     4021 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     4081 cagtacatca agtgtatcat atgccaagtc cgccccctat tgacgtcaat gacggtaaat
     4141 ggcccgcctg gcattatgcc cagtacatga ccttacggga ctttcctact tggcagtaca
     4201 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac accaatgggc
     4261 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     4321 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaataac cccgccccgt
     4381 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag
     4441 tgaaccgtca gatcactaga agctttattg cggtagttta tcacagttaa attgctaacg
     4501 cagtcagtgc ttctgacaca acagtctcga acttaagctg cagaagttgg tcgtgaggca
     4561 ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct
     4621 tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc
     4681 actttgcctt tctctccaca ggtgtccact cccagttcaa ttacagctct taaggctaga
     4741 gtattaatac gactcactat agggctagcg atcgccatgg cggcggccga cggcgacgac
     4801 tcgctgtacc ccatcgcggt gctcatagac gaactccgca atgaggacgt tcagcttcgc
     4861 ctcaacagca tcaagaagct gtccaccatc gccttggccc ttggggttga aaggacccga
     4921 agtgagcttc tgcctttcct tacagatacc atctatgatg aagatgaggt cctcctggcc
     4981 ctggcagaac agctgggaac cttcactacc ctggtgggag gcccagagta cgtgcactgc
     5041 ctgctgccac cgctggagtc gctggccaca gtggaggaga cagtggtgcg ggacaaggca
     5101 gtggagtcct tacgggccat ctcacacgag cactcgccct ctgacctgga ggcgcacttt
     5161 gtgccgctag tgaagcggct ggcgggcggc gactggttca cctcccgcac ctcggcctgc
     5221 ggcctcttct ccgtctgcta cccccgagtg tccagtgctg tgaaggcgga acttcgacag
     5281 tacttccgga acctgtgctc agatgacacc cccatggtgc ggcgggccgc agcctccaag
     5341 ctgggggagt ttgccaaggt gctggagctg gacaacgtca agagtgagat catccccatg
     5401 ttctccaacc tggcctctga cgagcaggac tcggtgcggc tgctggcggt ggaggcgtgc
     5461 gtgaacatcg cccagcttct gccccaggag gatctggagg ccctggtgat gcccactctg
     5521 cgccaggccg ctgaagacaa gtcctggcgc gtccgctaca tggtggctga caagttcaca
     5581 gagctccaga aagcagtggg gcctgagatc accaagacag acctggtccc tgccttccag
     5641 aacctgatga aagactgtga ggccgaggtg agggccgcag cctcccacaa ggtcaaagag
     5701 ttctgtgaaa acctctcagc tgactgtcgg gagaatgtga tcatgtccca gatcttgccc
     5761 tgcatcaagg agctggtgtc cgatgccaac caacatgtca agtctgccct ggcctcagtc
     5821 atcatgggtc tctctcccat cttgggcaaa gacaacacca tcgagcacct cttgcccctc
     5881 ttcctggctc agctgaagga tgagtgccct gaggtacggc tgaacatcat ctctaacctg
     5941 gactgtgtga acgaggtgat tggcatccgg cagctgtccc agtccctgct ccctgccatt
     6001 gtggagctgg ctgaggacgc caagtggcgg gtgcggctgg ccatcattga gtacatgccc
     6061 ctcctggctg gacagctggg agtggagttc tttgatgaga aacttaactc cttgtgcatg
     6121 gcctggcttg tggatcatgt atatgccatc cgcgaggcag ccaccagcaa cctgaagaag
     6181 ctagtggaaa agtttgggaa ggagtgggcc catgccacaa tcatccccaa ggtcttggcc
     6241 atgtccggag accccaacta cctgcaccgc atgactacgc tcttctgcat caatgtgctg
     6301 tctgaggtct gtgggcagga catcaccacc aagcacatgc tacccacggt tctgcgcatg
     6361 gctggggacc cggttgccaa tgtccgcttc aatgtggcca agtctctgca gaagataggg
     6421 cccatcctgg acaacagcac cttgcagagt gaagtcaagc ccatcctaga gaagctgacc
     6481 caggaccagg atgtggacgt caaatacttt
//
