LOCUS       sequence-425759-        1765 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Fluorescent reporter for glucosyl-ceramide (putative, mammalian 
            expression).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1765)
  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 1765)
  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- sfGFP-KSR1-CA3  (N-KSR)
FEATURES             Location/Qualifiers
     source          1..1765
                     /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             436..453
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             511..1221
                     /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             1519..1542
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1579..1602
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1629..1646
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1699..1717)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1713..1759
                     /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 gatccctggt tccacgtggt agcaactccg ctgttgatgg taccgcaggt
      481 ccgggttcta ttgcgactat ggtcgaattc tctaaaggcg aggaactgtt tacgggcgtc
      541 gttccaatcc tggtagaact ggatggtgac gtgaacggcc acaaattttc cgtacgtggt
      601 gaaggcgaag gcgatgcaac caacggtaaa ctgaccctca aatttatctg caccactggc
      661 aaactgccgg ttccgtggcc aaccctggtg accaccctga cttacggtgt acagtgtttc
      721 tctcgttatc cggaccatat gaaacgtcac gactttttca aaagcgcaat gccggagggt
      781 tacgttcagg aacgtacgat ttctttcaaa gacgacggca cgtacaaaac tcgcgctgaa
      841 gtgaaattcg aaggcgatac cctggttaac cgcatcgagc tgaaaggcat cgatttcaaa
      901 gaggacggca acatcctggg tcacaaactg gaatacaact tcaactctca taacgtgtat
      961 atcaccgcgg ataaacagaa aaacggcatc aaagccaact tcaaaatccg tcacaacgtg
     1021 gaagacggtt ctgtacagct ggcggaccac tatcagcaga acaccccaat tggtgacggt
     1081 ccggttctgc tgccagataa ccattacctg agcactcagt ccgtgctgtc caaagatccg
     1141 aacgaaaaac gcgatcacat ggtcctcctg gagttcgtta ctgccgctgg tattacccac
     1201 ggtatggacg aactgtacaa aggatccggc ccacagggcc tggggaaccg cattgatgac
     1261 gtctcctcga tgaggtttga tctctcgcat ggatccccac agatggtacg gagggatatc
     1321 gggctgtcgg tgacgcacag gttctccacc aagtcctggc tgtcgcaggt ctgccacgtg
     1381 tgccagaaga gcatgatatt tggagtgaag tgcaagcatt gcaggttgaa gtgtcacaac
     1441 aaatgtacca aagaagcccc tgcctgtaga atatccttcc tgccactaac tcggtaaggc
     1501 catatcggcc ttaagctttg gagccacccg cagttcgaga aaggtggagg ttccggaggt
     1561 ggatcgggag gtggatcgtg gagccacccg cagttcgaaa aagcggccga ttaagcggcc
     1621 gcctcgagca ccaccaccac caccactgag atccggctgc taacaaagcc cgaaaggaag
     1681 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaacg
     1741 ggtcttgagg ggttttttgc tgaaa
//
