LOCUS       sequence-153402-         859 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Positive control plasmid for Nicking Mutagenesis. Constitutively 
            expresses a non-fluorescent variant of GFPmut3 (Y66H) that is 
            mutated to fluorescent (H66Y) in the protocol..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 859)
  AUTHORS   Wrenbeck EE, Klesmith JR, Stapleton JA, Adeniran A, Tyo KE, 
            Whitehead TA
  TITLE     Plasmid-based one-pot saturation mutagenesis.
  JOURNAL   Nat Methods. 2016 Nov;13(11):928-930. doi: 10.1038/nmeth.4029. Epub 
            2016 Oct 10.
  PUBMED    27723752
REFERENCE   2  (bases 1 to 859)
  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"; doi: 
            "10.1038/nmeth.4029"; journalName: "Nat Methods"; date: "2016-11"; 
            volume: "13"; issue: "11"; pages: "928-930"
COMMENT     Sequence Label: pEDA5_GFPmut3_Y66H
FEATURES             Location/Qualifiers
     source          1..859
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             333..355
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(393..421)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
ORIGIN
        1 accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc
       61 tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc
      121 ctcaacgaca ggagcacgat catgcgcgct gaggacccgt ggggccgcca tgccggccac
      181 agctaacacc acgtcgtccc tatctgctgc cctaggtcta tgagtggttg ctggataact
      241 ttacgggcat gcataaggct cgtaatatat attcagggag accacaacgg tttccctcta
      301 caaataattt tgtttaactt tctagaaata attttgttta actttaagaa ggagatatac
      361 atatgcgtaa aggagaagaa cttttcactg gagttgtccc aattcttgtt gaattagatg
      421 gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat gcaacatacg
      481 gaaaacttac ccttaaattt atttgcacta ctggaaaact acctgttcca tggccaacac
      541 ttgtcactac tttcggtcat ggtgttcaat gctttgcgag atacccagat catatgaaac
      601 agcatgactt tttcaagagt gccatgcccg aaggttatgt acaggaaaga actatatttt
      661 tcaaagatga cgggaactac aagacacgtg ctgaagtcaa gtttgaaggt gatacccttg
      721 ttaatagaat cgagttaaaa ggtattgatt ttaaagaaga tggaaacatt cttggacaca
      781 aattggaata caactataac tcacacaatg tatacatcat ggcagacaaa caaaagaatg
      841 gaatcaaagt taacttcaa
//
