LOCUS       sequence-91547-e        1010 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 1010)
  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 1010)
  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..1010
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     7..23
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7..23
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             34..54
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             61..771
                     /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"
     CDS             772..795
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(825..841)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(826..842)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(860..878)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
ORIGIN
        1 ccggattcta gaactagtgg atcccccccc ggggaaaatt tatattttca aggtcaattt
       61 tctaaaggtg aagaattatt cactggcgtt gtcccaattt tggttgaatt agatggtgat
      121 gttaatggtc acaaattttc tgtctccggt gaaggtgaag gtgacgctac ttacggtaaa
      181 ttgaccttaa aatttatttg tactactggt aaattgccag ttccatggcc aaccttagtc
      241 actactttcg gttatggtgt tcaatgtttt gcgagatacc cagatcatat gaaacaacat
      301 gactttttca agtctgccat gccagaaggt tatgttcaag aaagaactat ttttttcaaa
      361 gatgacggta actacaagac cagagctgaa gtcaagtttg aaggtgatac cttagttaat
      421 agaatcgaat taaaaggtat tgattttaaa gaagatggta acattttagg tcacaaattg
      481 gaatacaact ataactctca caatgtttac atcatggctg acaaacaaaa gaatggtatc
      541 aaagttaact tcaaaattag acacaacatt gaagatggtt ctgttcaatt agctgaccat
      601 tatcaacaaa atactccaat tggtgatggt ccagtcttgt taccagacaa ccattactta
      661 tccactcaat ctgccttatc caaagatcca aacgaaaaga gagaccacat ggtcttgtta
      721 gaatttgtta ctgctgctgg tattacccat ggtatggatg aattgtacaa acaccaccac
      781 catcatcatc atcattaact gcaggaattc gatatcaagc ttatcgatac cgtcgacctc
      841 gagtcatgta attagttatg tcacgcttac attcacgccc ctccccccac atccgctcta
      901 accgaaaagg gaaggagtta gacaacctga agtctaggtc ccctatttat ttttttatag
      961 ttatgttagt attaagaacg ttatttatat tcaaattttc ctttttttcc
//
