LOCUS       sequence-224362-        2196 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Mammalian expression of the AKAR5 sensor tagged with nucleus export 
            signal (NES) 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 2196)
  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 2196)
  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_cyt-AKAR5
FEATURES             Location/Qualifiers
     source          1..2196
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     10..32
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     94..113
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      183..192
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(189..191,192..194,195..905)
                     /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(1428..1430,1431..1433,1434..2144)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNWNSVNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     ERVTAAGITLGMDELYK"
ORIGIN
        1 gcagggcggg gttcggcttc tggcgtgtga ccggcggctc tagagcctct gctaaccatg
       61 ttcatgcctt cttctttttc ctacagctcc tgggcaacgt gctggttatt gtgctgtctc
      121 atcattttgg caaagaattc ggtaccgcgg gcccgggatc caccggaagc cacctgcatg
      181 cggccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
      241 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
      301 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
      361 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
      421 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
      481 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
      541 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
      601 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
      661 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
      721 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
      781 acaaccacta cctgagcacc cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc
      841 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
      901 acaagcgcat gcataagttt tctcaagaac agatcggcga aaacattgtg tgcagggtca
      961 tttgtaccac gggtcaaatt cccatccgag atttgtcagc tgatatttca caagtgctta
     1021 aggaaaaacg atccataaag aaagtttgga catttggtag aaacccagcc tgtgactatc
     1081 atttaggaaa catttcaaga ctgtcaaata agcatttcca aatactacta ggagaagacg
     1141 gtaacctttt attgaatgac atttccacta atgggacctg gttaaatggg caaaaagtcg
     1201 agaagaacag caatcagtta ctgtctcaag gtgatgaaat aaccgttggt gtaggcgtgg
     1261 aatcagatat tttatctctg gtcattttca taaacgacaa atttaagcag tgcctcgagc
     1321 agaacaaagt tgatcgctct gcaggtaagc caggcagcgg cgagggcagc accaagggcc
     1381 tgcgtcgcgc caccctggtg gacggcggca ccggcggcag cgagctcatg gtgagcaagg
     1441 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     1501 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     1561 tgaagctcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct
     1621 tcggctacgg cctgatgtgc ttcgcccgct accccgacca catgaagcag cacgacttct
     1681 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     1741 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     1801 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     1861 actggaacag cgtcaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     1921 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     1981 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc
     2041 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagagag
     2101 tgaccgccgc cgggatcact ctcggcatgg acgagctata caagggtgaa ttccttcaaa
     2161 aaaaacttga agaacttgaa cttgacgaat gatcta
//
