LOCUS       sequence-165809-        2257 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses GFP-Tagged OSBP PH domain in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2257)
  AUTHORS   Levine TP, Munro S
  TITLE     The pleckstrin homology domain of oxysterol-binding protein 
            recognises a determinant specific to Golgi membranes.
  JOURNAL   Curr Biol. 1998 Jun 18;8(13):729-39.
  PUBMED    9651677
REFERENCE   2  (bases 1 to 2257)
  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: "Curr 
            Biol."; date: "1998-06-18"; volume: "8(13)"; pages: "729-39"
COMMENT     Sequence Label: pRS406-PHO5-GFP-OSBP PH domain
FEATURES             Location/Qualifiers
     source          1..2257
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     225..247
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     239..256
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     240..256
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     266..285
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        266..284
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     513..532
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             797..1510
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(827..855)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1450..1470
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             1523..1552
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(1897..1917)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1897..1915)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1936..1952)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1936..1952)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1949..1971)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1960..1976
                     /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)."
     promoter        complement(join(1984..1990,1991..2008,2009..2014))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2029..2050
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2167..2184)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
       61 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc cattcgccat tcaggctgcg
      121 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      181 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      241 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg
      301 gccccccctc gaggtcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt
      361 agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg
      421 cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg
      481 agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca
      541 accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgatccg
      601 gtcaccttac ttggcaaggc atatacccat ttgggataag ggtaaacatc tttgaattgt
      661 cgaaatgaaa cgtatataag cgctgatgtt ttgctaagtc gaggttagta tggcttcatc
      721 tctcatgaga ataagaacaa caacaaatag agcaagcaaa ttcgagaatt cctgcagccc
      781 gggcaattcg aatataatgt ctaaaggaga agaacttttc actggagttg tcccaattct
      841 tgttgaatta gatggtgatg ttaatgggca caaattttct gtcagtggag agggtgaagg
      901 tgatgcaaca tacggaaaac ttacccttaa atttatttgc actactggaa aactacctgt
      961 tccatggcca acacttgtca ctacttttac gtatggtgtt caatgctttt caagataccc
     1021 agatcatatg aaacggcatg actttttcaa gagtgccatg cccgaaggtt atgtacagga
     1081 aagaactata tttttcaaag atgacgggaa ctacaagaca cgtgctgaag tcaagtttga
     1141 aggtgatacc cttgttaata gaatcgagtt aaaaggtatt gattttaaag aagatggaaa
     1201 cattcttgga cacaaattgg aatacaacta taactcacac aatgtataca tcatggcaga
     1261 caaacaaaag aatggaatca aagttaactt caaaattaga cacaacattg aagatggaag
     1321 cgttcaacta gcagaccatt atcaacaaaa tactccaatt ggcgatggcc ctgtcctttt
     1381 accagacaac cattacctgt ccacacaatc tgccctttcg aaagatccca acgaaaagag
     1441 agaccacatg gtccttcttg agtttgtaac agctgctggg attacacatg gcatggatga
     1501 actatacaag cttggatcca tggagcaaaa gctcatttct gaagaggact tgagatcttc
     1561 gggctcggct cgagagggct ggctcttcaa atggaccaat tatatcaaag gctaccagcg
     1621 gcgatggttc gtgctgagca acgggctcct gagctactac agatcaaagg cagagatgag
     1681 acatacctgc cgtggtacca tcaacctcgc cacagccaac atcaccgtgg aggactcctg
     1741 caacttcatc atttccaatg ggggtgctca gacctaccat ctgaaagcta gttcagaagt
     1801 tgagcggcag cgctgggtga cggccctgga actggccaag gccaaagctg tgaagaagaa
     1861 ttcgtgataa ccgcggtgga gctccagctt ttgttccctt tagtgagggt taattgcgcg
     1921 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     1981 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     2041 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     2101 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     2161 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     2221 tcactcaaag gcggtaatac ggttatccac agaatca
//
