LOCUS       sequence-102875-         907 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  FRET-based biosensor for monitoring cAMP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 907)
  AUTHORS   DiPilato LM, Zhang J
  TITLE     The role of membrane microdomains in shaping beta2-adrenergic 
            receptor-mediated cAMP dynamics.
  JOURNAL   Mol Biosyst. 2009 Aug;5(8):832-7. doi: 10.1039/b823243a. Epub 2009 
            May 26.
  PUBMED    19603118
REFERENCE   2  (bases 1 to 907)
  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: "10.1039/b823243a"; 
            journalName: "Mol Biosyst"; date: "2009-08"; volume: "5"; issue: 
            "8"; pages: "832-7"
COMMENT     Sequence Label: pcDNA3-ICUE3
FEATURES             Location/Qualifiers
     source          1..907
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     36..55
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        36..54
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      74..83
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             95..112
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             116..148
                     /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(152..160,161..175)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             join(182..184,185..187,188..898)
                     /codon_start=1
                     /product="monomeric super enhanced variant of CFP (Matsuda 
                     et al., 2008)"
                     /label=mseCFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     CDS             join(182..184,185..187,188..898)
                     /codon_start=1
                     /product="cyan variant of GFP"
                     /label=CFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     CDS             join(182..184,185..187,188..898)
                     /codon_start=1
                     /product="enhanced CFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(227..248)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(488..507)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     835..856
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 actagagaac ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc
       61 caagcttgcg gccgccacca tggtgcgggg ttctcatcat catcatcatc atggtatggc
      121 tagcatgact ggtggacagc aaatgggtcg ggatctgtac gacgatgacg ataaggatcc
      181 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
      241 cggcgacgta aacggccaca ggttcagcgt gtccggcgag ggcgagggcg atgccaccta
      301 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
      361 cctcgtgacc accctgacct ggggcgtgca gtgcttcagc cgctaccccg accacatgaa
      421 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gtaccatctt
      481 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
      541 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
      601 caagctggag tacaactaca tcagccacaa cgtctatatc accgccgaca agcagaagaa
      661 cggcatcaag gcccacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
      721 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
      781 ctacctgagc acccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt
      841 cctgctggag ttcgtgaccg ccgccgggat cactcacggc atggacgagc tgtacaaggc
      901 tggtacc
//
