LOCUS       sequence-43796-e         964 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 964)
  AUTHORS   Hsieh JM, Besserer GM, Madej MG, Bui HQ, Kwon S, Abramson J
  TITLE     Bridging the gap: a GFP-based strategy for overexpression and 
            purification of membrane proteins with intra and extracellular 
            C-termini.
  JOURNAL   Protein Sci. 2010 Apr;19(4):868-80.
  PUBMED    20196076
REFERENCE   2  (bases 1 to 964)
  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."; date: "2010-04"; volume: "19(4)"; pages: "868-80"
COMMENT     Sequence Label: pWarf(+)
FEATURES             Location/Qualifiers
     source          1..964
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             50..73
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             191..901
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKAN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(218..246)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     841..861
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             914..937
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
ORIGIN
        1 actttaagaa ggagactcga gatgttaaga attccgggta cctggatcct tggaagtctt
       61 gtttcaggga ccattaatta acggtggtga gattacatta attatttttg gtgtgatggc
      121 tggtgttatt ggaacgatcc tcttaatttc ttacggtatt cgtcgtctga ttaagggttc
      181 tggtggtttc agcaaaggag aagaactttt cactggagtt gtcccaattc ttgttgaatt
      241 agatggtgat gttaatgggc acaaattttc tgtcagtgga gagggtgaag gtgatgctac
      301 atacggaaaa ctcaccctta aatttatttg cactactgga aaactacctg ttccatggcc
      361 aacacttgtc actactctga cctatggtgt tcaatgcttt tcccgttatc cggatcacat
      421 gaaacggcat gactttttca agagtgccat gcccgaaggt tatgtacagg aacgcactat
      481 atctttcaaa gatgacggga actacaagac gcgtgctgaa gtcaagtttg aaggtgatac
      541 ccttgttaat cgtatcgagt taaagggtat tgattttaaa gaagatggaa acattctcgg
      601 acacaaacta gagtacaact ataactcaca caatgtatac atcacggcag acaaacaaaa
      661 gaatggaatc aaagctaact tcaaaattcg ccacaacatt gaagatggtt ccgttcaact
      721 agcagaccat tatcaacaaa atactccaat tggcgatggc cctgtccttt taccagacaa
      781 ccattacctg tcgacacaat ctgccctttc gaaagatccc aacgaaaagc gtgaccacat
      841 ggtccttctt gagtttgtaa ctgctgctgg gattacacat ggcatggatg agctctacaa
      901 aaagcttgcg gcccatcatc atcaccacca ccaccactga gatccggctg ctaacaagcc
      961 cgaa
//
