LOCUS       sequence-91544-e        1045 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Used for expressing recombinant proteins as a C terminal GFP-His 
            tagged constuct in S.cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1045)
  AUTHORS   Parker JL, Newstead S
  TITLE     Method to increase the yield of eukaryotic membrane protein 
            expression in Saccharomyces cerevisiae for structural and functional
            studies.
  JOURNAL   Protein Sci. 2014 Jun 20. doi: 10.1002/pro.2507.
  PUBMED    24947543
REFERENCE   2  (bases 1 to 1045)
  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: "Protein 
            Sci. 2014 Jun 20. doi: 10.1002/pro.2507."
COMMENT     Sequence Label: pDDGFP_LEU2D
FEATURES             Location/Qualifiers
     source          1..1045
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     terminator      complement(13..202)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     214..232
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     250..266
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     251..267
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(297..320)
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             complement(321..1031)
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
ORIGIN
        1 gcgcaattaa gccttcgagc gtcccaaaac cttctcaagc aaggttttca gtataatgtt
       61 acatgcgtac acgcgtctgt acagaaaaaa aagaaaaatt tgaaatataa ataacgttct
      121 taatactaac ataactataa aaaaataaat agggacctag acttcaggtt gtctaactcc
      181 ttccttttcg gttagagcgg atgtgggggg agggcgtgaa tgtaagcgtg acataactaa
      241 ttacatgact cgaggtcgac ggtatcgata agcttgatat cgaattcctg cagttaatga
      301 tgatgatgat ggtggtggtg tttgtacaat tcatccatac catgggtaat accagcagca
      361 gtaacaaatt ctaacaagac catgtggtct ctcttttcgt ttggatcttt ggataaggca
      421 gattgagtgg ataagtaatg gttgtctggt aacaagactg gaccatcacc aattggagta
      481 ttttgttgat aatggtcagc taattgaaca gaaccatctt caatgttgtg tctaattttg
      541 aagttaactt tgataccatt cttttgtttg tcagccatga tgtaaacatt gtgagagtta
      601 tagttgtatt ccaatttgtg acctaaaatg ttaccatctt ctttaaaatc aatacctttt
      661 aattcgattc tattaactaa ggtatcacct tcaaacttga cttcagctct ggtcttgtag
      721 ttaccgtcat ctttgaaaaa aatagttctt tcttgaacat aaccttctgg catggcagac
      781 ttgaaaaagt catgttgttt catatgatct gggtatctcg caaaacattg aacaccataa
      841 ccgaaagtag tgactaaggt tggccatgga actggcaatt taccagtagt acaaataaat
      901 tttaaggtca atttaccgta agtagcgtca ccttcacctt caccggagac agaaaatttg
      961 tgaccattaa catcaccatc taattcaacc aaaattggga caacgccagt gaataattct
     1021 tcacctttag aaaattgacc ttgaa
//
