LOCUS       sequence-355638-        2520 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Replacement of endogenous rat PKA-RII-alpha subunits with wild-type 
            RII-alpha.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2520)
  AUTHORS   Church TW, Tewatia P, Hannan S, Antunes J, Eriksson O, Smart TG, 
            Hellgren Kotaleski J, Gold MG
  TITLE     AKAP79 enables calcineurin to directly suppress protein kinase A 
            activity.
  JOURNAL   Elife. 2021 Oct 6;10. pii: 68164. doi: 10.7554/eLife.68164.
  PUBMED    34612814
REFERENCE   2  (bases 1 to 2520)
  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"; journalName: "Elife."; 
            date: "2021-10-6"; volume: "10. pii: 68164. doi"; pages: " 
            10.7554/eLife.68164"
COMMENT     Sequence Label: pFUGW-shRII-alpha-RII-alpha-WT-IRES-EGFP
FEATURES             Location/Qualifiers
     source          1..2520
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_feature    join(1217..1791,1792..1794,1795..1803)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1384..1401)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1611..1630
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1804..1806,1807..1809,1810..2520)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1804..1806,1807..1809,1810..2520)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1849..1870)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2110..2129)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2457..2478
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 atgagccaca tccagatccc accggggctc acggagctgc tgcagggcta caccgtggag
       61 gtgcttcggc agcagccgcc cgacctcgtc gacttcgcgg tggagtactt cacacgcctg
      121 cgcgaggccc gccggcaaga gtcagactcg ttcatcgccc ccccgacgac ctttcacgcg
      181 caggagtcca gcggggtccc cgtcatcgag gaggacgggg agagtgaatc ggactcggac
      241 gatgaggatc tggaagttcc gattcccagc aaatttacta gacgagtatc agtctgtgca
      301 gaaacgttta accctgatga agaagaagat aatgatccaa gggtggttca ccccaaaacc
      361 gacgagcaga ggtgcagact tcaggaagcc tgtaaagaca ttctgctttt caaaaacctg
      421 gatcaggaac agctttctca agttctggac gccatgtttg aaaagatagt caaaactgac
      481 gagcatgtca ttgaccaagg agatgatgga gacaactttt atgtcataga aaggggaacc
      541 tatgacattt tagtaacaaa agataatcaa acacgatctg ttggtcagta tgacaaccgt
      601 ggcagttttg gagaactagc cctgatgtac aataccccga gagctgctac cattgtggcc
      661 acctcagacg gctccctttg gggattggac cgggtgactt ttaggagaat catagtgaag
      721 aacaatgcaa agaagaggaa gatgttcgaa tcgtttattg agtctgtacc gctctttaaa
      781 tcactagaga tgtcagaacg aatgaagatt gtggatgtga tcggggaaaa gatctataag
      841 gatggagagc gaataatcac tcagggtgaa aaggccgaca gcttttatat tatagagtct
      901 ggagaagtga gcatcttgat tagaagcaag actaaaacga acaagaacgg cgggaaccag
      961 gaggttgaga ttgcccactg ccataagggg cagtactttg gagaacttgc cctggtgacc
     1021 aacaaaccaa gagctgcttc tgcttatgcg gttggagacg tcaaatgctt agtcatggat
     1081 gttcaagcat ttgagaggct tctgggcccc tgcatggaca tcatgaagag gaacatctca
     1141 cattacgaag aacagctggt gaagatgttt ggctccaact tggatctatt ggaccccggg
     1201 cagtagctcg gatccgcccc tctccctccc ccccccctaa cgttactggc cgaagccgct
     1261 tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg
     1321 gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt
     1381 cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg
     1441 aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac
     1501 ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg
     1561 cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct
     1621 caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg
     1681 atctggggcc tcggtacaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag
     1741 gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa tatggccaca
     1801 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     1861 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     1921 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     1981 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     2041 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     2101 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     2161 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     2221 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     2281 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     2341 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     2401 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     2461 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
//
