LOCUS       sequence-425272-        2069 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Fluorescent reporter for ceramide (putative, mammalian expression).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2069)
  AUTHORS   Girik V, van Ek L, Dentand Quadri I, Azam M, Cruz Cobo M, Mandavit 
            M, Riezman I, Riezman H, Gavin AC, Nunes-Hasler P
  TITLE     Development of Genetically Encoded Fluorescent KSR1-Based Probes to 
            Track Ceramides during Phagocytosis.
  JOURNAL   Int J Mol Sci. 2024 Mar 5;25(5):2996. doi: 10.3390/ijms25052996.
  PUBMED    38474242
REFERENCE   2  (bases 1 to 2069)
  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: "Int J Mol 
            Sci."; date: "2024-03-5"; volume: "25(5):2996. doi"; pages: " 
            10.3390/ijms25052996"
COMMENT     Sequence Label: pEGFP-PRKCZ-C20 (N-PRKCZ-C20)
FEATURES             Location/Qualifiers
     source          1..2069
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        230..433
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     384..404
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      472..481
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(478..480,481..483,484..1194)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(478..480,481..483,484..1194)
                     /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(523..544)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(784..803)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1131..1152
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1948..2069
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1985..2004)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2039..2058
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
       61 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
      121 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
      181 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
      241 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
      301 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
      361 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
      421 tataagcaga gctggtttag tgaaccgtca gatccgctag cgctaccggt cgccaccatg
      481 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
      541 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
      601 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
      661 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
      721 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
      781 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
      841 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
      901 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
      961 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     1021 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     1081 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     1141 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtccgga
     1201 ctcagatctc gagctggcgg ttcctctggt ggtggtggtg cgggtgacac aacgagcact
     1261 ttctgcggaa ccccgaatta catcgccccc gaaatcctgc ggggagagga gtacgggttc
     1321 agcgtggact ggtgggcgct gggagtcctc atgtttgaga tgatggccgg gcgctccccg
     1381 ttcgacatca tcaccgacaa cccggacatg aacacagagg actacctttt ccaagtgatc
     1441 ctggagaagc ccatccggat cccccggttc ctgtccgtca aagcctccca tgttttaaaa
     1501 ggatttttaa ataaggaccc caaagagagg ctcggctgcc ggccacagac tggattttct
     1561 gacatcaagt cccacgcgtt cttccgcagc atagactggg acttgctgga gaagaagcag
     1621 gcgctccctc cattccagcc acagatcaca gacgactacg gtctggacaa ctttgacaca
     1681 cagttcacca gcgagcccgt gcagctgacc ccagacgatg aggatgccat aaagaggatc
     1741 gaccagtcag agttcgaagg ctttgagtat atcaacccat tattgctgtc caccgaggag
     1801 tcgggtaccg cgggcccggg atccaccgga tctagataac tgatcataat cagccatacc
     1861 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     1921 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     1981 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     2041 ggtttgtcca aactcatcaa tgtatctta
//
