LOCUS       sequence-224366-        2213 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Mammalian expression of the AKAR5 sensor C-terminally fused to the 
            C-terminus of KRas (membrane targeting) 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 2213)
  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 2213)
  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_m-AKAR5
FEATURES             Location/Qualifiers
     source          1..2213
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     73..92
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      162..171
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(168..170,171..173,174..884)
                     /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(1407..1409,1410..1412,1413..2123)
                     /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 ggcgtgtgac cggcggctct agagcctctg ctaaccatgt tcatgccttc ttctttttcc
       61 tacagctcct gggcaacgtg ctggttattg tgctgtctca tcattttggc aaagaattcg
      121 gtaccgcggg cccgggatcc accggaagcc acctgcatgc ggccaccatg gtgagcaagg
      181 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
      241 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
      301 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
      361 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
      421 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
      481 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
      541 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
      601 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
      661 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
      721 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
      781 agtccaagct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
      841 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagcgcatg cataagtttt
      901 ctcaagaaca gatcggcgaa aacattgtgt gcagggtcat ttgtaccacg ggtcaaattc
      961 ccatccgaga tttgtcagct gatatttcac aagtgcttaa ggaaaaacga tccataaaga
     1021 aagtttggac atttggtaga aacccagcct gtgactatca tttaggaaac atttcaagac
     1081 tgtcaaataa gcatttccaa atactactag gagaagacgg taacctttta ttgaatgaca
     1141 tttccactaa tgggacctgg ttaaatgggc aaaaagtcga gaagaacagc aatcagttac
     1201 tgtctcaagg tgatgaaata accgttggtg taggcgtgga atcagatatt ttatctctgg
     1261 tcattttcat aaacgacaaa tttaagcagt gcctcgagca gaacaaagtt gatcgctctg
     1321 caggtaagcc aggcagcggc gagggcagca ccaagggcct gcgtcgcgcc accctggtgg
     1381 acggcggcac cggcggcagc gagctcatgg tgagcaaggg cgaggagctg ttcaccgggg
     1441 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     1501 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagctcatc tgcaccaccg
     1561 gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt cggctacggc ctgatgtgct
     1621 tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     1681 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     1741 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     1801 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctggaacagc gtcaacgtct
     1861 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     1921 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     1981 gccccgtgct gctgcccgac aaccactacc tgagctacca gtccgccctg agcaaagacc
     2041 ccaacgagaa gcgcgatcac atggtcctgc tggagagagt gaccgccgcc gggatcactc
     2101 tcggcatgga cgagctatac aagggtgaat tcggtaaaaa gaagaaaaag aagtcaaaga
     2161 caaagtgtgt aattatgtaa tctagagtcg aggtaccgat gaatagctaa ggt
//
