LOCUS       sequence-224353-        2189 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Mammalian expression of the AKAR5' sensor for detecting PKA activity
            using two-photon fluorescence lifetime imaging microscopy (2pFLIM).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2189)
  AUTHORS   Tillo SE, Xiong WH, Takahashi M, Miao S, Andrade AL, Fortin DA, Yang
            G, Qin M, Smoody BF, Stork PJS, Zhong H
  TITLE     Liberated PKA Catalytic Subunits Associate with the Membrane via 
            Myristoylation to Preferentially Phosphorylate Membrane Substrates.
  JOURNAL   Cell Rep. 2017 Apr 18;19(3):617-629. doi: 
            10.1016/j.celrep.2017.03.070.
  PUBMED    28423323
REFERENCE   2  (bases 1 to 2189)
  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"; doi: 
            "10.1016/j.celrep.2017.03"; journalName: "Cell Rep"; date: 
            "2017-04-18- 18"; volume: "19"; issue: "3"; pages: "617-629"
COMMENT     Sequence Label: pCAGGS_AKAR5'
FEATURES             Location/Qualifiers
     source          1..2189
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     65..84
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      154..163
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(160..162,163..165,166..876)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1615..1617,1618..1620,1621..2133)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNWNSVNVYIMADKQKNGIK
                     VNFKIRHNIE"
ORIGIN
        1 accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc
       61 ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt cggtaccgcg
      121 ggcccgggat ccaccggaag ccacctgcat gcggccacca tggtgagcaa gggcgaggag
      181 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
      241 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
      301 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
      361 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
      421 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
      481 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
      541 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
      601 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
      661 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
      721 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccaag
      781 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
      841 gccgggatca ctctcggcat ggacgagctg tacaagcgca tgcataagtt ttctcaagaa
      901 cagatcggcg aaaacattgt gtgcagggtc atttgtacca cgggtcaaat tcccatccga
      961 gatttgtcag ctgatatttc acaagtgctt aaggaaaaac gatccataaa gaaagtttgg
     1021 acatttggta gaaacccagc ctgtgactat catttaggaa acatttcaag actgtcaaat
     1081 aagcatttcc aaatactact aggagaagac ggtaaccttt tattgaatga catttccact
     1141 aatgggacct ggttaaatgg gcaaaaagtc gagaagaaca gcaatcagtt actgtctcaa
     1201 ggtgatgaaa taaccgttgg tgtaggcgtg gaatcagata ttttatctct ggtcattttc
     1261 ataaacgaca aatttaagca gtgcctcgag cagaacaaag ttgatcgctc tgcaggtaag
     1321 ccaggcagcg gcgagggcag caccaagggc ctgcgtcgcg ccaccctggt ggacggcggc
     1381 accggcggca gcgagctcat gggcagcgtg cagctcgccg accactacca gcagaacacc
     1441 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccgcc
     1501 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagag agtgaccgcc
     1561 gccgggatca ctctcggcat ggacgagcta tacaagggtg gaactggagg ttcaatggtg
     1621 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     1681 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     1741 ctgaccctga agctcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     1801 accaccttcg gctacggcct gatgtgcttc gcccgctacc ccgaccacat gaagcagcac
     1861 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     1921 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     1981 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     2041 gagtacaact ggaacagcgt caacgtctat atcatggccg acaagcagaa gaacggcatc
     2101 aaggtgaact tcaagatccg ccacaacatc gagtaaggcc gcatgatcag ctggatgcat
     2161 cgatcacgcg taccggtgct cgaggtacc
//
