LOCUS       sequence-425273-        2190 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 2190)
  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 2190)
  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: pNES-PRKCZ-C20-EGFP (C-NES-PRKCZ-C20)
FEATURES             Location/Qualifiers
     source          1..2190
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      537..546
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    1161..1213
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      1221..1230
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1227..1229,1230..1232,1233..1946)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1227..1229,1230..1232,1233..1943)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1272..1293)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1533..1552)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1880..1901
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2069..2190
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2106..2125)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2160..2179
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcca
      541 ccatggctct gcagaaaaag ttggaagagc ttgagctgga tgaggcaccg gtcgccaccg
      601 gtgacacaac gagcactttc tgcggaaccc cgaattacat cgcccccgaa atcctgcggg
      661 gagaggagta cgggttcagc gtggactggt gggcgctggg agtcctcatg tttgagatga
      721 tggccgggcg ctccccgttc gacatcatca ccgacaaccc ggacatgaac acagaggact
      781 accttttcca agtgatcctg gagaagccca tccggatccc ccggttcctg tccgtcaaag
      841 cctcccatgt tttaaaagga tttttaaata aggaccccaa agagaggctc ggctgccggc
      901 cacagactgg attttctgac atcaagtccc acgcgttctt ccgcagcata gactgggact
      961 tgctggagaa gaagcaggcg ctccctccat tccagccaca gatcacagac gactacggtc
     1021 tggacaactt tgacacacag ttcaccagcg agcccgtgca gctgacccca gacgatgagg
     1081 atgccataaa gaggatcgac cagtcagagt tcgaaggctt tgagtatatc aacccattat
     1141 tgctgtccac cgaggagtcg ctcgagctca agcttcgaat tctgcagtcg acggtaccgc
     1201 gggcccggga tccaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg
     1261 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     1321 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     1381 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     1441 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     1501 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     1561 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     1621 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     1681 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     1741 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     1801 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     1861 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     1921 tcggcatgga cgagctgtac aagtaaagcg gccgcgactc tagatcataa tcagccatac
     1981 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     2041 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     2101 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     2161 tggtttgtcc aaactcatca atgtatctta
//
