LOCUS       sequence-425760-        1597 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Fluorescent reporter for ceramide (for bacterial expression and 
            purification).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1597)
  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 1597)
  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: pETM11-SUMO3-KSR1-CA3-sfGFP (C-KSR)
FEATURES             Location/Qualifiers
     source          1..1597
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     24..43
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        24..42
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    43..67
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             118..135
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             166..186
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             730..1443
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     CDS             1461..1478
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1531..1549)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1545..1591
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tcgagatctc gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa
       61 caattcccct ctagaaataa ttttgattta actttaagaa ggagatatac catgaaacat
      121 caccatcacc atcaccccat gagcgattac gacatcccca ctactgagaa tctttatttt
      181 cagggcgcca tgggcaacga tcacattaac ctgaaagtgg ccggtcaaga cggtagcgta
      241 gtccagttta aaatcaaacg ccacacccct ctgtcgaaac tgatgaaagc ctattgtgaa
      301 cgccaaggtc tgtctatgcg tcagatccgt tttcgcttcg atggacagcc gattaacgaa
      361 accgacactc cagcacagct ggaaatggaa gatgaggaca ccattgacgt gttccagcaa
      421 cagaccggtg gatccaccat ggggaaccgc attgatgacg tctcctcgat gaggtttgat
      481 ctctcgcatg gatccccaca gatggtacgg agggatatcg ggctgtcggt gacgcacagg
      541 ttctccacca agtcctggct gtcgcaggtc tgccacgtgt gccagaagag catgatattt
      601 ggagtgaagt gcaagcattg caggttgaag tgtcacaaca aatgtaccaa agaagcccct
      661 gcctgtagaa tatccttcct gccactaact cgggcaggtc cgggttctat tgcgactatg
      721 gtcgaattct ctaaaggcga ggaactgttt acgggcgtcg ttccaatcct ggtagaactg
      781 gatggtgacg tgaacggcca caaattttcc gtacgtggtg aaggcgaagg cgatgcaacc
      841 aacggtaaac tgaccctcaa atttatctgc accactggca aactgccggt tccgtggcca
      901 accctggtga ccaccctgac ttacggtgta cagtgtttct ctcgttatcc ggaccatatg
      961 aaacgtcacg actttttcaa aagcgcaatg ccggagggtt acgttcagga acgtacgatt
     1021 tctttcaaag acgacggcac gtacaaaact cgcgctgaag tgaaattcga aggcgatacc
     1081 ctggttaacc gcatcgagct gaaaggcatc gatttcaaag aggacggcaa catcctgggt
     1141 cacaaactgg aatacaactt caactctcat aacgtgtata tcaccgcgga taaacagaaa
     1201 aacggcatca aagccaactt caaaatccgt cacaacgtgg aagacggttc tgtacagctg
     1261 gcggaccact atcagcagaa caccccaatt ggtgacggtc cggttctgct gccagataac
     1321 cattacctga gcactcagtc cgtgctgtcc aaagatccga acgaaaaacg cgatcacatg
     1381 gtcctcctgg agttcgttac tgccgctggt attacccacg gtatggacga actgtacaaa
     1441 tgataagcgg ccgcctcgag caccaccacc accaccactg anatccggct gctaacaaag
     1501 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
     1561 gggcctctaa cgggtcttga ggggtttttt gctgaaa
//
