LOCUS       sequence-100398-        1074 bp    DNA     linear   UNA 14-APR-2025
DEFINITION  A fluorescent reporter of AMPK activity and cellular energy stress.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1074)
  AUTHORS   Tsou P, Zheng B, Hsu CH, Sasaki AT, Cantley LC
  TITLE     A fluorescent reporter of AMPK activity and cellular energy stress.
  JOURNAL   Cell Metab. 2011 Apr 6;13(4):476-86.
  PUBMED    21459332
REFERENCE   2  (bases 1 to 1074)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell 
            Metab."; date: "2011-04-6"; volume: "13(4)"; pages: "476-86"
COMMENT     Sequence Label: ampkar
FEATURES             Location/Qualifiers
     source          1..1074
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     61..80
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        61..79
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      99..108
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             120..137
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             141..173
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(177..185,186..200)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             join(207..209,210..212,213..725)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIE"
     primer_bind     complement(252..273)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(513..532)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     860..881
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 ctatataagc agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat
       61 taatacgact cactataggg agacccaagc ttgcggccgc caccatggtg cggggttctc
      121 atcatcatca tcatcatggt atggctagca tgactggtgg acagcaaatg ggtcgggatc
      181 tgtacgacga tgacgataag gatcccatgg tgagcaaggg cgaggagctg ttcaccgggg
      241 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaggttc agcgtgtccg
      301 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
      361 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctggggc gtgcagtgct
      421 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
      481 gctacgtcca ggagcgtacc atcttcttca aggacgacgg caactacaag acccgcgccg
      541 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
      601 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacatcagc cacaacgtct
      661 atatcaccgc cgacaagcag aagaacggca tcaaggccca cttcaagatc cgccacaaca
      721 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
      781 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccaagctg agcaaagacc
      841 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc cgcatgcata
      901 agttttctca agaacagatc ggcgaaaaca ttgtgtgcag ggtcatttgt accacgggtc
      961 aaattcccat ccgagatttg tcagctgata tttcacaagt gcttaaggaa aaacgatcca
     1021 taaagaaagt ttggacattt ggtagaaacc cagcctgtga ctatcattta ggaa
//
