LOCUS       sequence-103169-         878 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  PCR template for en passant (two-step Red-mediated) mutagenesis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 878)
  AUTHORS   Tischer BK, von Einem J, Kaufer B, Osterrieder N
  TITLE     Two-step red-mediated recombination for versatile high-efficiency 
            markerless DNA manipulation in Escherichia coli.
  JOURNAL   Biotechniques. 2006 Feb;40(2):191-7.
  PUBMED    16526409
REFERENCE   2  (bases 1 to 878)
  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: 
            "Biotechniques."; date: "2006-02"; volume: "40(2)"; pages: "191-7"
COMMENT     Sequence Label: pEPkan-S2
FEATURES             Location/Qualifiers
     source          1..878
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     47..66
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        47..65
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             97..120
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(251..270)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(335..354)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 ctctctggct aactagagaa cccactgctt actggcttat cgaaattaat acgactcact
       61 atagggagac ccaagcttgg taccgagctc ggatccgatt acaaggatga cgacgataag
      121 tagggataac agggtaatcg atttattcaa caaagccacg ttgtgtctca aaatctctga
      181 tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata
      241 aacagtaatg caaggggtgt tatgagccat attcaacggg aaacgtcttg ctcgaggccg
      301 cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc
      361 gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc agagttgttt
      421 ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac
      481 tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat
      541 gcatggttac tcaccactgc gatccccggg aaaacagcat tccaggtatt agaagaatat
      601 cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg
      661 attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa
      721 tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg
      781 cctgttgaac aagtctggaa agaaatgcat aagcttttgc cattctcacc ggattcagtc
      841 gtcactcatg gtgatttctc acttgataac cttatttt
//
